Annals of the Missouri Botanical Garden Volume Ш 1916 With Nine Piates opd Eighty-four Figures Published quarterly by the Board of Trustees of the Missouri Botanical Garden, St. Louis, Mo. Entered as second-class matter at the Post Office at St. Louis, Missouri, under the Act of March 3, 1879. 24088 Annals of the Missouri Botanical Garden A Quarterly Journal containing Scientifie Contributions from the Missouri Botanical Garden and the Graduate Labora- tory of the Henry Shaw School of Botany of Washington University in affiliation with the Missouri Botanical Garden. Editorial Committee George T. Moore Benjamin M. Duggar Information The Annals of the Missouri Botanical Garden appears four times dur- ing the calendar year, February, April, September, and November. Four numbers constitute a volume. Subscription Price - - $3.00 per volume Single Numbers - - - 1.00 each The following agent is authorized to aceept foreign subseriptions: William Wesley & Son, 28 Essex Street, Strand, London. \ "^ d. . . ° ein $8 Seier Pria | NA Мат СА А d'et cos 9: Рту D e... e STAFF OF THE MISSOURI BOTANICAL GARDEN Director, GEORGE T. MOORE. BENJAMIN М, DUGGAR, EDWARD A. Burt, Physiologist, in We - of Mycologist and Librarian. Graduate Laborator J. C. Тн. UPHOF, HERMANN VON Me 5 TA Assistant Botanist. Patholog JESSE M. GREENMAN, GEORGE W. FREIBERG, Curator of the Herbarium. Research Assistant. KATHERINE H. LEIGH, Secretary to the Director. BOARD OF TRUSTEES OF THE MISSOURI BOTANICAL GARDEN President, EDWARDS WHITAKER. Vice-President, DAVID S. H. SMI EDWARD ©, ELIOT. LEONARD MATTHEWS. GEORGE C. HITCHCOCK. WILLIAM H. H. PETTUS. P. CHOUTEAU MAFFITT. PHILIP C. SCANLAN. EDWARD MALLINCKRODT. JOHN F. SHEPLEY. EX-OFFICIO MEMBERS: EDMUND A. ENG HENRY W. KIE President of the жы of Science Mayor of the eU of St. Louis. uis. BENJAMIN P. STROMBER Еверекіс A. HALL President of the Board of Public Chancellor of Washington University. Schools of St. Lou DANIEL Š. TUTTLE Bishop of the Diocese of Missouri. E. F. FINNEY, Assistant Secretary. : 2, Gë е _— ba : с. TABLE OF CONTENTS Rhizoctonia Solani in Relation to the “МорорП2” and the ‘‘ Vermehrung- BUR ce › os scm ue SER a ae B. M. Duggar The Texas Root Rot Fungus and Its Conidial Stage: З EE ee B. M. Duggar Cabbage Yellows and the Relation of Temperature to Its Occurrence..... J. C. Gilman Monograph of the North and Central American Species of the Genus Senecio—Part II............... J. M. Greenman New or Interesting Species of Gill Fungi from Missouri............ L. O. Overholts A New Senecio from Jamaica..... J. M. Greenman The Thelephoraceae of North America. Vv prec cer ee TTE E. A. Burt The Occurrence in Nature of Certain Yeast-like Fungi with Reference to Their Possible Pathogenicity in the E EE W. H. Emig The Missouri Agrimonies............. B. F. Bush The Thelephoraceae of North America. VELO. МЕ s E. A. Burt Catalogue of the Plants of Jasper County, Miageouriy. Zen | за E. J. Palmer Pistillaria (subg. Pistillina) Thaxteri, БЕРЕКЕСІ CG AX | ane cue ke E. A. Burt PAGE 85-194 195-200 201-202 203—241 243-307 309-318 319-343 345-401 403-406 TABLE OF CONTENTS PAGE A Note on the Adaptability of the Folin Micro-Kjeldahl Apparatus for Plant SN Ee ЕТТЕ АК A.R. Davis 407-412 Studies in the Physiology of the Fungi. I. Nitrogen Fixation............... B. M. Duggar and A. R. Davis 413-437 Studies in the Physiology of the Fungi. II. Lenzites saepiaria Fries, with Special Reference to Enzyme Ас- КҮЙЕДІ ыты т S. M. Zeller 489-512 General Index to Volume ПІ..................... 513-517 Annals of the Missouri Botanical Garden Vor. 3 FEBRUARY, 1916 No. 1 RHIZOCTONIA SOLANI IN RELATION TO THE * MOPOPILZ" AND THE *VERMEHRUNGSPILZ " B. M. DUGGAR Physiologist to the Missouri Botanical Garden, in Charge of Graduate Laboratory Professor of Plant EC in the Henry Shaw School of Botany of Washington Universi Discussing in a recent paper (15) the distribution of Rhiz- octonia Solani Kühn (Corticium vagum B. & C.) I made the following statements: “И is rather surprising to find that R. Solani has received relatively little attention in Europe. Although recognized as inducing a disease of the potato widely distributed in central Europe, and occasionally reported on the beet, yet little careful work has been bestowed upon the fungus.” T shall now endeavor to show, as definitely as a discussion of the literature will permit, that this statement requires ma- terial modification. At the same time the evidence indicates a considerable extension of the region in which this Rhizoc- tonia is important as a seed-bed parasite. The new light on the problem is a result of the provisional determination— amounting almost to a certainty—that the Javanese ‘‘Mo- popilz" and the central European ‘‘Vermehrungspilz’’ are identical with Rhizoctonia Solani (Corticium vagum B. & C.). Some years ago the writer attempted to determine the pos- sible relation of Rhizoctonia to seed-bed or cutting-bench dis- eases in Germany, but at that time the literature was scant and confusing, so that the effort was unfortunately abandoned, largely, ee er, as a result of the suggestions from several sourees, that Pythium, Botrytis, and other known forms were Ann. Mo. Bor. Garb., VoL. 3, 1916 (1) [VoL. 3 2 ANNALS OF THE MISSOURI BOTANICAL GARDEN clearly responsible for these diseases. It now appears that a considerable literature has been gradually accumulating, but it was not correlated with the work on Rhizoctonia, proceed- ing at the same time, owing largely to incorrect determina- tions. I owe the present outlook upon the literature to the illuminating paper of Rant (715%) in which, it would seem, from his description and figures, that he clearly and correctly identifies the ‘‘mopo’’ fungus of Cinchona seed-beds in Java as Momliopsis Aderholdu, described by Ruhland (208) and designated by him as the cause of seed-bed and propagation- bed difficulties. It was necessary for me to go but a step further to determine that Moniliopsis Aderholdii is in reality identical with Rhizoctonia Solani. It seems well, however, to review briefly all the contributions thus far found which seem to shed light on this fungus as a cause of disease in propa- gating-beds, as studied in Europe and in Java, especially as it serves to supplement the literature cited in my recent paper (Duggar, 715). Since comments will be made in eonneetion with the re- view of literature, it may be well to emphasize certain char- acteristics particularly distinctive of Rhizoctonia Solani, and among those important are the following: (1) The great variety of higher plants affected; (2) the rapidity of spread where seedlings are attacked, presenting an appearance as if hot water had been poured over the young plants; (5) the growth of a web of mycelium over the fallen plants and likewise over the adjacent soil, so that frag- ments of soil adhere when the plants are lifted 3 (4) mycelium practically hyaline when young, with characteristic branching and septation, becoming brownish with age; (5) under favor- able conditions, especially in culture, the development of floc- cose masses, consisting of chains of cells (Monilia-like), often much branched or elbowed, colorless to brownish; (6) the for- mation of dark brown sclerotial bodies, irregular in size and outline, developing in the same way as the Йоссове masses, but denser by anastomosis, with the form of the cells (in mature E practically uniform throughout, that is, with no dif- s perhaps more than any other characteristic enables нин to dis- Der the fungus from the effects of Pythium and of Botryti 1916] DUGGAR—RHIZOCTONIA SOLANI 3 ferentiated peripheral layer; and (7) cultures readily ob- tained from mycelium or sclerotium, the organism producing in culture only mycelium, flake-like masses or tufts, and ef- fuse sclerotia. In reviewing the earlier of these studies upon diseases of this type which may be caused by Rhizoctonia it is hazardous to attempt to interpret those cases in which the organism is in- adequately described, yet bearing in mind the more striking characteristics of Rhizoctonia, it is believed that no litera- ture is here included which does not suggest this fungus. In the later studies the organism has been for the most part so well described that little doubt may be entertained with re- spect to the determination. For the present it is necessary to rely upon a discussion of the literature, but when more ma- terial, in the form of cultures, from the regions here referred to is available, a supplementary statement will be required. An examination of the files of the more important of the horticultural journals of both France and Germany prior to 1880, has been made with the result that references to dis- eases of cuttings and seedlings are found to be not infrequent, but without exception these contribute nothing, so far as I have been able to find, which will throw light upon the organ- ism concerned. The earliest reference which has been found to be of importance is that of Therry and Thierry ('82). They reported having studied, for more than a year, the my- celial filaments which invaded Ше cutting-benches of garden- ers and florists in the region of Lyons. Although unable to find a spore stage, they described the organism studied as Mortierella arachnoides Th. & Th. (araignée des serres), since they considered the vegetative stage to show a close re- lationship to M. Ficariae which they found on leaves of Ficaria Ranunculoides. M. arachnoides is described as killing the shoots and growing over the fallen tissues, disorganizing them with great rapidity, also growing over the soil in the form of a web of strands. The mycelium is said to be able to grow meters during a single night. The points noted, together with the brief description of the mycelium, strongly sug- Tü ae a on [VoL. 3 4 ANNALS OF THE MISSOURI BOTANICAL GARDEN gest Rhizoctonia, and it would not be strange that sclerotia were absent under the conditions. Prompted apparently by the account just referred to, and based somewhat upon that, von Thümen (’82) reports upon the ‘‘Vermehrungspilz,’’ and this appears to be the first defi- nite account of the organism from central Europe. His de- scription of the mycelium adds somewhat to that of Therry and Thierry, and like them he found that “* * * die Unter- suchungen nichts weiter ergab, als die Anwesenheit zahl- reicher, spinnwebendünne, weisslicher oder braunlicher My- celfüden, von irgend einer Fruetifieation aber trotz genaues- ten Suchens auch nicht die mindeste Spur aufzufinden мат.” Whatever may be the interpretation of these two sets of ob- servations they emphasize (1) the rapidity of growth and vio- lence of the attacks of the organism concerned and (2) the presence of a mycelium as the only stage of the associated organism. It appears probable that the disease which came to be known as ‘‘maladie de la toile?" in France is the same as that re- ferred to by Therry and Thierry (282); nevertheless, such ob- servations as are reported during the next fifteen years leave the question of a causal organism in an unsatisfactory state. Mangin (794) refers to ‘‘la toile" as the disease due to a fun- gus occurring both in the greenhouse and in the open, pro- ducing a decay of leaves and branches, especially at or near the surface of the soil. Recalling what has already been said regarding this fungus it is significant that he remarks: “Quand la Toile est bien développée, les filaments mycéliens agglutinent les fragments de terre et deviennent trés visibles." Collecting material from the affected area he found that in a few days conidiophores of Botrytis appeared on the dead leaves. With cultures of the Botrytis he reproduced a dis- ease in lettuce. Since, however, Botrytis cinerea might oc- eur upon any debris, and since it also produces a disease of lettuce, it does not follow, of course, that it is the fungus re- sponsible for the troubles here referred to. From the de- scription of the effects, one is inclined to reject the idea that Botrytis is concerned in this ease. In the same year Prillieux 1916] DUGGAR—RHIZOCTONIA SOLANI 5 and Delacroix (794) found “Ча toile’’ abundant in the seed and propagating-beds, market gardens, ete., near Fontaine- bleau. Affected plants were infested with a sterile mycelium, and they found a fungus, identified as Botrytis cinerea, fruit- ing on the dead material, from which they prepared cultures. No infection experiments were made, and they report no at- tempt to ascertain if the mycelium in the tissues were really that of Botrytis. No additional information is advanced in Mangin's (794*) second paper. Sorauer (796) refers to the ‘‘ Vermehrungsschimmel" of the cutting-benches and of seed-beds as probably belonging to the genus Sclerotinia. Reference is made to the spider web-like mycelium, lack of sporophores, and the presence of sclerotia. It is apparently on account of the sclerotia that he refers the fungus to Sclerotinia. He indieates that this organism is the chief fungus of the cutting-bench, although Mucor, Botrytis, and other organisms may also be found. The description, though far from being complete, is applicable to Rhizoctonia. Aderhold (797) characterized the fungus and its effects in some detail, and there ean be little doubt that he was dealing with the disease then recognized as widely distributed. More- over, unlike those who preceded him, he obtained the sterile fungus in culture, observed the Monilia-like chains of cells, and also the formation of sclerotia. It seems remarkable that it did not suggest to him Kühn's potato fungus. On the con- trary, he agreed with Sorauer in referring the fungus to Sclerotinia, without indieating the species. In a second paper Sorauer (299) also discusses the fungus more completely. He refers to much of the earlier work, in- cluding that of Aderhold. Various stages of the fungus are figured, that is, the mycelium, the moniliform hyphal cells, and the sclerotia, all stages pointing to Rhizoctonia. He also re- fers to a characteristic of his fungus, since frequently ob- served, doubtless, by all who have studied Rhizoctonia in liquid eultures, namely, that of growing up the walls of the vessel above the level of the liquid. He also examined the affected tissues and was able to follow the mycelium in its advance, showing its penetration into the inner bark, like- [Vor. 8 6 ANNALS OF THE MISSOURI BOTANICAL GARDEN wise into the mesophyll of affected leaves. Comment is made on the fact that the death of the cells ensues when very few hyphae have penetrated the tissues. A review of the earlier work on ‘‘la toile" in France is presented by Beauverie (’99) who calls attention to the fact that the fungus producing this disease has been considered by some to be Botrytis cinerea, and by others to be Acros- talagmus albus, a determination made in one instance at least by Oudemans (’92). This determination was based on mate- rial received from the Zodlogical Garden at Rotterdam. Beauverie obtained cultures but fails to describe how the fun- gus was isolated. From these cultures he was able to obtain only a sterile fungus, which, unfortunately, is not described. Failing to obtain spores he then proceeded with cultures origi- nating from Botrytis cinerea. By growing this organism in a moist atmosphere at a temperature of about 33°C. a sterile form was induced. Again, by exposing cultures to a tempera- ture somewhat lower, he affirms that he was able to develop a temporary sterile stage. It would appear that on the basis of these results he draws the conclusion that the first organism isolated represented also a sterile form of the Botrytis. He further emphasizes the point that the sterile form is the more dangerous in the production of disease, leading to the infer- ence that conditions resulting in the development of this stage predetermined the prevalence of the malady. It is unsatisfac- tory to attempt to draw conclusions from this work, but it is at least probable that his first cultures may have been Rhi- госіота, and that, however accurately the work with Botry- tis may have been carried out, it had really no connection with ‘Ја toile.’’ Lindau (’08) follows his discussion of Rhizoctonia with a paragraph dealing with the fungus producing disease in the eutting-beneh and propagating-houses. The organism is de- seribed on the basis of the observations of Sorauer (799) and Aderhold (297), reference being made to the characteristic mycelium and chains of short cells, as well as to the occurrence of sclerotia. He questions the relationship with Monilia, sug- 1916] DUGGAR—RHIZOCTONIA SOLANI 7 gesting that the figures would indicate a closer relationship to Hormiscium or Torula. Since many of the American stud- ies upon the potato and damping off fungus had previously been examined by him, as the account of Rhizoctonia indicates, it is surprising that the possibility of the identity of the ‘‘Vermehrungspilz’’ with Rhizoctonia was not suggested. In his discussion of the fungus Ruhland (208) considers the earlier work of Aderhold, Beauverie, and Sorauer. Special attention is given to that of Beauverie, and Ruhland takes the view that while in all probability the disease discussed by that investigator is the same as the disease of propagating-beds in Germany, Beauverie's cultures of Botrytis were not those of the disease-inducing organism. Ruhland studied the or- ganism in culture, confirms the previous descriptions of my- celium, Monilia-like cells, structure of the sclerotia, ete. He would regard the sclerotia as sclerotial-like bodies (pseudo- sclerotia), owing to the fact that the structure is homogeneous throughout. Cultures of the fungus here discussed and of Botrytis cinerea were studied in parallel series with respect to the capacity to ferment cellulose, and it was found that while this capacity is possessed to a considerable degree by Botrytis, as had been previously established, the cellulose-dis- solving capacity of the **Vermehrungspilz" is very low. He finds that in the development of the Monilia-like cells there is only a superficial resemblance to Monilia, since the spores of the latter are produced basipetally, while those of the seed- bed fungus are formed aeropetally. The development of scle- rotia from the Monilia-like masses is also noted. Apparently, he concluded that the old cells of the Monilia-like chains, as well as those of the sclerotia, were empty, hence incapable of germination. Earlier studies of Rhizoctonia have, however, shown clearly that many old cells of this type are capable of germination, and the peculiarities of this process have been figured and described (Duggar, '99). In Java a disease of the Cinchona seed-beds was reported by Moens (789). He describes the damping off of the seedlings as rapidly progressing radially, especially when the condi- tions are moist. 'The disease often begins at those points [Vor. 3 8 ANNALS OF THE MISSOURI BOTANICAL GARDEN where the drip from the defective roof falls on the seed-bed. The mycelium is described as spreading rapidly in the form of a web over the diseased plants and adjacent soil. Some observations were made by Stibbe (’06) who also reported that the disease may appear as early as during the first few days of growth. Koorders (706) observed the disease in a young plantation. An examination of affected stems and roots was made, and a colorless, septate mycelium was found, but there were no evidences of fruiting stages. From these earlier observations of the Cinchona diseases in Java we have only the above evidences of the effect upon the host to suggest Rhizoctonia or a related fungus. The investigations of Rant (208, 714, 715, '15*), previously referred to, are sufficiently complete in all partieulars to en- able us to identify the fungus as Rhizoctonia. To this dis- ease he applies the term ‘торо’? and ‘‘hamamopo’’ rather than the Dutch ‘‘Schimmeldraadjes.’’ He found the effects upon the host to be as previously described, and noted particu- larly that the cobweb-like growth of the mycelium over the soil and dead plants occurred in a characteristic fashion when the area over the seed-bed was moist. He also emphasizes the point that fragments of soil are firmly held together by the growth of the fungous mycelium. Referring again to the distinctive characteristics of Rhizoctonia enumerated earlier, we find that his work covers every point there indicated. The fungus was found to affect not only Cinchona seedlings but was also found in his garden upon the following: Lychnis diurna, Rudbeckia sp., Lobelia erinus, Conyza angustifolia, Bidens pilosa, Antirrhinum majus, red beet, endive, cabbage, and lettuce. Culturing the fungus upon peptone glucose agar he obtained a good growth with all the characteristics of Rhi- zoctonia which have been referred to in my previous paper. Comparing his measurements with those previously reported, it is found that there is a close agreement throughout. The measurements also agree with those of Aderhold (’97). Rant also instituted a comparison between this fungus and Botry- tis cinerea, and the results well emphasize the differences be- tween these two organisms. He found likewise that the 1916] DUGGAR—RHIZOCTONIA SOLANI 9 ‘‘mopo’’ fungus bears no resemblance to Acrostalagmus al- bus, to which “Ча toile" in France had been occasionally re- ferred. On the other hand, cultures of the ‘‘Vermehrung- spilz’’ obtained from Amsterdam agreed closely with the or- ganism isolated from Cinchona seedlings. Infection experi- ments with Ше “торо” fungus were carried out both on Cin- chona seedlings and on seedlings of the plants previously enumerated as affected. In all cases positive results were ob- tained. Summary and Discussion—From the reviews and discus- sion it seems justifiable to conclude that the common seed- bed fungus in Germany and in France is identical with the damping off fungus which has been frequently studied in this country since the investigations of Atkinson (’92). I have given the evidence upon which the conclusion is based that the damping off fungus of the United States is Kiihn’s Rhi- zoctoma Solani (Corticium vagum B. & C.), the cause of im- portant potato diseases and of other types of disease in a variety of host plants. The work of Rant enables us to in- clude in this category of diseases due to Rhizoctonia Solani the disease of Cinchona seedlings and of other plants in Java. In establishing these points it is not necessary to consider the earlier and less complete reports upon ‘Іа toile’? and the ‘‘Vermehrungspilz’’ but particularly the papers of Aderhold (797), Sorauer (799), Beauverie (99), and Ruhland (’08). LITERATURE CITED Aderhold, R. ('97). Über den АА чече == sein Leben und seine Bekümp- ung. Gartenflora 46:114—126. f. 1-5. n Ge C'90). Lg имене Esp eott IV. ‘‘Sore-shin,’’ арии EL ра rot." Agr. ваг Bul. 41:30-39. KE Beauverie, J. (09). Le Botrytis Шеге et la maladie de la toile. apt rend. Acad. Paris 128:846-849, 1251-1253. 1899. 250. 1909 Duggar, B. M. (799). Three important fungous diseases of the sugar beet. Cornell Univ. Agr. Exp. Sta., Bul. 163:339-363. f. 49-63. 1899. резки, ДКА et eg A. (’09). Maladies parasitaires des plantes cultivées. РР. СТБ are cae EE (Pers.) DC. and R. Solani Kühn (Cor tieium vagum B. & C.), h notes on other ee Ann. Mo. Bot. Gard. 2:403-458. f. 1- -9. 1915. [Vou. 3 10 ANNALS OF THE MISSOURI BOTANICAL GARDEN кишер 8. H. (’06). Resultaten van een voorloopig mikroskopisch onderzoek жен van jonge kinaplantjes veroorzaakt door Heterodera-aaltjes sehimmel. De Cultuurgids. 1906. [Cited from Rant, Lendner, A. (’14). Une maladie de la vigne due à un champignon du genre Hypoehnus. Soc. Bot. Genéve, Bul. II. 6:104—106. 1914. we Eé à (708). (In Sorauer's) Handbuch der Pflanzenkrankheiten 2:473-474. 190 ет L. (794). Sur la toile, oo parasitaire de certains végétaux. Bul. Soe. Biol. X. 1:209-210. 189 (7942). Sur le parasitisme d'une espéce de Botrytis. Compt. rend. Acad. Paris 118':882-884. 1894. Moens, J. C. B. (782). Die Kinaeultuur in Azié 1882. pp. 152-153. 1882. Oudemans, C. A. J. A. ( 99). Gergen A la A mycologique des Pays-Bas. XIV. Nederl. Krudk. Arch. II. 6:p. 1892. Plemper van Balen, B. A. (700). Het ‘‘omvallen’’ van stekken. Tijdsch. Plan- tenziekten 6: 30- 81. 1900. Prillieux, E., E Delacroix, С. (794). Maladie de la toile produite par le Botrytis ompt. rend. Acad. Paris 118:744-746. 1894. Rant, A. (708). De Mopoziekte. Teysmannia 19:433-434. 1908. — — ——, (214). Die Ziekten en Schimmels der Kina. Mededeelingen van het Kina Proefstation. 1914. [Cited from "Rant, "15а. | —— (15). De Mopoziekte. Teysmannia 26:54-57. 1915. (7152). Uber die Mopokrankheit junger Cinehonapflanzen und über den "javanischen Vermehrungspilz. Jard. Bot. Buitenzorg, Bul. 18:1-21. f. 1—8. 1915. сети T (708). "ar v Kenntnis m sog. CERS “6; biol. Anst. f. Land- u. rstw., Arb. 6:71-76. f. 1-3. 1908 — p (796). Über б eet bte Pflanzenkr. 4-315. 1896. , (99). Der Vermehrungspilz. Ibid. 9:321-328. pl. 6. 1899. Stibbe, A. F. W. ('06). Eenige aanvullingen op de voordracht van den heer V. W. van Goch, gehouden op het 6de Congres te Malang. De Cultuurgids. 1906. [Cited from Rant, ’ Therry, J., et Thierry (782). Nouvelles be Am de Mucorinées du genre Mor- tierella. Rev. Myc. 4:160-162. 1882 Thiimen, F. v. (’82). Der at PD Wiener Garten Zeit. 7:417-419. 1882. THE TEXAS ROOT ROT FUNGUS AND ITS CONIDIAL STAGE! B. M. DUGGAR Physiologist to the Missouri Botanical Garden, in Charge of Graduate Laboratory Professor of Plant Physiology in the Henry Shaw School of Botany of Washington University More than twenty-five years ago Pammel ('88, '89) spent two summers in Texas investigating an important cotton dis- ease popularly known as the ‘‘cotton blight’’ or “cotton dy- ing,” and as а result of his observations two reports were published upon the cotton root rot,—the latter more appro- priate name being applied by him to the disease in question. He determined the causal organism to be a sterile fungus found in some abundance on every dead or dying root, and it was tentatively identified as Ozonium auricomum Lk. After a study of Link’s type, Shear (707) described the organism as a new species, O. omnivorum. Since the work of Pammel the disease has doubtless been the object of numerous field observations, more or less exten- sive, but so far as is personally known to me, and so far as reports are available, the only records are those of Dug- gar (709, observations made 701, ’02), Shear (707, observa- tions beginning in '02), Shear and Miles (207, 707%), Heald (209, 711), and Heald and Wolf (711, 19). The disease is un- doubtedly one of the most destructive of the cotton fungi, and the average losses sustained in the state of Texas have been variously estimated by Orton ('06) and others to be two to three million dollars. In addition, considerable damage is sustained by such erops as alfalfa, beans, sweet potatoes, and certain orchard fruits. It would appear that the organism is very largely confined їо Texas. In that state it seems to have been commonly ob- "The writer was engaged in a study of cotton diseases, Ди зе тъй Ше Texas root rot, in Ше Bur reau ы Plant Industry during the seasons of 1901-02. In the fall of 1902 the work was transferred to Dr. C. L. Shear. Now that the Missouri Botanical Garden is giving ple cie attention to a botanieal survey a certain sec- tion of the Southwest, it has seemed appropriate to resume the studies of this fungus so wide-spread and pe espinas, in a large part of that region. Ann. Mo. Bor. Garp., Vor. 3, 1916. (11) ve dA ы. e [Vor. 8 12 ANNALS OF THE MISSOURI BOTANICAL GARDEN served since 1866. Shear (’07) states that the fungus is dis- tributed from eastern Texas to southern California, and that it has been found in southern Oklahoma and Indian Territory. The writer failed to find the organism in western Louisiana and southwestern Arkansas in 1901, but it was observed in southern Oklahoma in 1915. I am unaware of the data on which the occurrence of the fungus westward to California is reported. Nevertheless, considering the long period of time during which the Ozoniwm has been a serious disease-inducing factor in Texas, it is rather remarkable that it has not been found in Louisiana and Mississippi. In these states the cot- ton wilt fungus, Fusarium vasinfectum, is well known, but the Ozonium has never been reported, so far as can be learned. It is almost impossible to assume that the fungus has not been distributed to these states through the various possible commercial channels; so that one is impelled to draw the in- ference that the establishment of the fungus farther east- ward is limited by climatic or soil factors. It should be recalled that Pammel reported the disease common throughout all sections of Texas in which cotton was grown, with the exception of Ше gulf prairie region and cer- tain alluvial soils. It seems now certain that there 1s no soil type in the cotton-producing section of the state which is free from the disease. Nevertheless, the percentage of loss has been invariably greater in the black prairie or blaek waxy soils, whether with or without outeroppings of rotten lime- stone. As noted later, the organism occurs on a number of native plants, both trees and herbs, but the observations thus far made give no clue as to whether or not it may be eonsid- ered endemie. I have been unable to secure data on the oc- currence of this fungus from Mexico southward. From the reports of Pammel, Heald, Heald and Wolf, and from my own observations, the following host plants may be enumerated. Trees and shrubs: Ulmus americana, Broussonetia papy- rifera, Morus alba, Ficus Carica, Acer saccharinum, Tilia americana, Fraxinus americana, Diospyros Kaki, Melia Aze- darach, Pyrus communis, P. Malus, Cydonia vulgaris, Robinia Pseudo-Acacia, Prunus Persica, P. sp. (cherry), and Hibiscus syriacus. 1916] DUGGAR—THE TEXAS ROOT ROT FUNGUS 13 Herbaceous plants: Beta vulgaris, Chenopodium sp., Cas- sia Tora, C. marilandica, Medicago sativa, Arachis hypogaea, Phaseolus vulgaris, Vigna sinensis, Linum rigidum, Croton spp., Euphorbia spp. (three), Sida spinosa, Hibiscus esculen- tus, Gossypium herbaceum, Petroselinum hortense, Ipomoea Batatas, Solanum rostratum, Ambrosia psilostachya, and Xanthium canadense. So far as I am aware, no special attempt has been made to determine all the species of wild herbaceous plants or forest trees affected. The enumeration of hosts given above makes it seem plausible, therefore, that few plants or crops may be free from the disease except the grains and other members of the grass family. At Petty, Texas, in September, 1901, the disease was found upon half a dozen species of weeds in a pasture, the sod of which could not have been disturbed for some years previous. The chief characteristic of the disease, as far as I have ob- served it on herbaceous plants, is the sudden wilting and dy- ing of the affected individuals. Occasionally a slight yellow- ing and unhealthy appearance is found to be due to an infec- tion which does not encircle the main root, and less frequently to the localization of the disease in a few of the larger primary root branches. The first ‘‘dying’’ of cotton is associated with the beginning of blossoming, or of boll formation, commonly from June to July; but Pammel reports one case in which the disease was observed May 6. If the fungus is responsible for injuries in the early stages of growth, then either such in- juries have been overlooked or have been ascribed to other causes. common with Rhizoctonia Crocorum the organism spreads radially, the rate of spread being most variable and, of course, governed by the conditions. The most rapid spread observed by the writer was in a field of irrigated al- falfa. The persistence of the larger ‘‘dead spots’’ season after season in much the same part of the field is accountable in large measure for the popular belief that these are ‘‘al- kali” spots. The progress of the disease from one year to another is best followed by observing a perennial crop such as alfalfa, in which case new infections are usually relatively few, whereas in a field grown two years or more to cotton [VoL. 8 14 ANNALS OF THE MISSOURI BOTANICAL GARDEN one notes the disappearance of some of the smaller spots of the previous year, and often the number of new infections is considerable. If diseased cotton stalks are left standing in the field, few or no evidences of the fungus are apparent on the roots the following March. However, some of the more interested growers claim to have observed mats of the fungus turned over by the plow when bedding the land. I have been unable to obtain such material for study. Ав already indicated, the reappearance of the larger spots, particularly, is a strong indication of the persistence of the mycelium in the soil. This leaves out of consideration the influence of the conidial stage, discussed below, in the persistence of the organism in the same area during successive years. On lifting wilted stalks of cotton, or stalks recently dead, it is found, from the most favorable material, that the roots are closely invested with a einnamon-buff! felt of hyphae in which strands are conspicuous. The fungus may involve the small- est rootlets, and in addition, the strands of hyphae penetrate the soil and apparently extend considerable distances. The larger soil strands are somewhat darker, often cinnamon- colored. In a badly infected area the strands of hyphae may be found in any lump of soil. Pammel describes the mycelium as brown in color, and Shear as ‘‘dirty yellow, whitish when young.’’ In the early stages of development on the host, I find the mycelium pale buff, becoming cinnamon-brown as strands are formed. In September, 1915, the conditions were particularly favor- able at Paris, Texas, for late-season infections, so that by ex- amining the roots of many plants taken at the periphery of a diseased area, but themselves apparently healthy, compara- tively early stages of infection were observed. In all cases a depression of the bark pointed out the area of penetration of the fungus on the main root. The observations also demon- strated clearly that the attack may be either what I shall designate centripetal or centrifugal. In the former case the infection converges upon the main roots from a few or many small laterals, while in the latter the main root may be com- *Ridgway’s ‘Color standards and nomenclature’ has been employed in the determination of all colors referred to in this paper. 1916] DUGGAR—THE TEXAS ROOT ROT FUNGUS 15 pletely encircled before the fungus extends to the branches. If recovery of affected stalks oceurs at all, it is usually by the produetion of very superficial laterals. I have not made a careful study of the distribution of hyphae in the various tissues, nor of the mechanism of pene- tration. From the variety of plants affected it may be in- ferred that direct infection by the hyphae is general. The presence of lentieels on the enlarged part of the root of cot- ton by midsummer may possibly be related to the greater susceptibility of this plant, and may also be a factor in de- termining the frequency of the centrifugal type. During the seasons of 1901-09 a careful search was made for spore stages of the Ozoniwm, and while several basid- lomycetous fungi were found on old cotton stalks in areas where cotton had died from the disease, still no case was observed which, upon careful examination, proved worthy of experimental study. In the examination made of such ma- terial special care was given to the characters of the myce- lium. However, while examining a semicircular area of dead cotton on the edge of a cotton field in 1902, my attention was caught by a buff-colored circular spot on the ground just out- side the cotton field in an area of grass and weeds wherein several of the latter had died from the Ozoniwm. The ma- terial observed proved to be an incrustation, or light powdery layer, of spores covering about one square foot in area. One small area of a few square inches only, considerably weath- ered, was found between the rows of cotton. Removing the soil with some of the spore material and making an examination under the hand lens it was found that strands of the Ozonium pervaded the whole mass, and thus there was presented the possibility of a spore form genetically connected with the Ozonium. Subsequently, the spore material was studied more carefully. At that time it was clear that strands of the Ozoniwm were present under the masses of. spores, but the observations afforded no evidenee of the method of spore production. The masses of small spheroidal spores formed a layer sometimes 3 mm. in thickness, and while the broken bits of hyphae observed resembled those of the zonium, no light was thrown on the relation of spores to mycelium. The conditions so much resembled those under Uh [VoL. š 16 ANNALS OF THE MISSOURI BOTANICAL GARDEN which oidial formation occurs in cultures of certain Pasidio- mycetes that I subsequently suggested the presence of such an oidial stage of this fungus (Duggar, '09). Owing to the transfer of the cotton disease work to Dr. С. L. Shear at this time the material was laid away, and not again examined un- til a recent reinvestigation of all material in my hands which might be considered related to Rhizoctonia. Then it was as- certained that the best packet of material collected in 1902 had not been studied—that from the area found between the rows of cotton. The reéxamination of this collection resulted Fig. 1. Phymatotrichum omnivorum: types of conidiophores and conidial production. in finding in some abundance the hyphae which bear the spores. It was furthermore ascertained that the spores were produced at first on the characteristic larger hyphae and on small branches of those hyphae which make up the strands of the fungus in the soil. Typically, the attached conidia were found in heads about short swollen, but not necessarily spherical, branches of the short-celled or strand hyphae. These branches were simple or forked, the forking being at irregular intervals, and occasionally branching was contin- ued from a swollen cell (fig. 1). The spores adhered some- Д 1916] DUGGAR—THE TEXAS ROOT ROT FUNGUS 17 what, but never in such masses as characterize certain species of Sporotrichum or Verticillium. In view of the importance of this observation, and failing to secure material from correspondents, a trip to Paris, Texas, was arranged in September, 1915, with the view of securing fresh material and of making further observations on the fungus in the field. The time selected proved favor- able, and an examination of the cotton fields in the vicinity of Paris revealed the Оготшт in unusual abundance. Nev- ertheless, ‘‘dead spots’’ in many fields were examined before the characteristic fruiting stage was found. Then it was located in some quantity in a ** dead spot’’ of about one acre in extent, occurring in a very rich, black waxy soil in a low- lying area of the field. In this area no less than a hundred or more of these conidial cir- cles were found. They varied in diameter from 3 to 30 cm. The majority of these were found in the furrows or ‘‘mid- dles’’ between the rows of cot- ton, yet they also occurred оп the ridge rows, and in seven cases they gleck diseased cotton stalks. In the latter cases, however, the strands Fig. 2. mM omnivorum: penetrating the spore-bearing large- celled hypha layer appeared to come from the soil in general rather than directly from the diseased root. In every case the typical color of the spore mass was light pinkish cinnamon, and in thin strata pinkish buff, fading some- what on drying. At that date the circular area consisted of a more or less perfect crust of spores sometimes broken or pow- dery. A few of the spore areas had become overgrown with such olive-green moulds as Cladosporium and Macrosporiwm. 2 [VoL. 3 18 ANNALS OF THE MISSOURI BOTANICAL GARDEN Just below the spore crust, especially towards the center, the typical cinnamon-buff strands of the Ozoniwm occurred in abundance. A similar type of mycelium also permeated the soil to a considerable extent in the immediate vicinity and often about the periphery of the spore-bearing area. The study of the collections made in 1915 emphasize the diversity in the form of hyphae as well as in the method of spore production. Although no circular areas were found in an early stage of development, yet some of the older ones yielded on the periphery material from which the method of spore formation could be followed. It would appear that a superficial growth of large, branched, almost hyaline hyphae is first formed (fig. 2), covering the surface with a delicate stratum. These hyphae are sometimes Rhizoctonia-like. They may also bear conidiophores at irregular intervals, the latter arising usually as short assurgent branches. These branches either produce conidia directly, or commonly after becoming variously forked (fig. 1). As further growth pro- ceeds, however, definite strands are developed, and then swollen branches from any cell of the strand may produce spores. Later the wave of spore production appears to in- volve practically the whole mycelium, and the conidia are found laterally distributed in various positions on the surfaces of both the strand and simple hyphae, so that in the end there is practically nothing left but a pulverulent mass consisting of the conidia and remains of the mycelium and strands. The conidia are sessile, but occasionally cells bearing conidia ex- hibit a somewhat roughened surface. The true character of the fungus cannot be determined unless one is careful to se- cure the youngest material available, that is, from near the margin of the spore area, or otherwise a spore-forming area in an early stage of development. The diverse characteristics of the mycelium, as found on the surface of the host and in the soil, may be briefly sum- marized as follows: (1) Large-celled type. Hyphae Rhizoctonia-like, often abundant on the margins of the conidial areas, measuring frequently 20и in diameter, with cross walls 60-120» apart. This type should also include some of the arachnoid mycelium 1916] DUGGAR—THE TEXAS ROOT ROT FUNGUS 19 on the surface of the roots, also those representing early stages of strand formation (fig. 2). (2) Strand hyphae. In these the individuality of the hyphae is practically lost, the strands being ultimately plectenchy- matic bands in which the individual cells vary considerably in diameter, the larger cells of young strands resembling some- what the larger hyphae above mentioned. It is interesting to observe that they may serve not only to spread the fungus vegetatively, but superficial soil strands may function as a conidial stroma. They are also more or less sclerotial and are doubtless an important factor in the persistence of the fungus in the soil (fig. 3). (3) Acicular type. The arachnoid mycelium with which the root is invested gives rise to certain fairly rigid hyphae AN GO оку Sen š EE E, SE E . 3. Phymatotrichum omnivorum: mycelial strands; upper, old strand from root of cotton; lower, young strand from conidial area. which in turn produce branches that are most frequently in pairs, that is, opposite, and at right angles. Branching is also not infrequently verticillate. In all cases such branches are characteristic in appearance, being rigid and needle-like, tapering to very fine filaments (fig. 4). This type has been found only on the roots. It is necessary to add, however, that intermediate types between the various forms mentioned occur. In general, the [VoL. 8 20 ANNALS OF THE MISSOURI BOTANICAL GARDEN mycelium is Rhizoetonia-like rather than Ozonium-like, yet no sclerotia have been found. In this connection I may add that it is proposed in a later paper to bring together certain observations which have been accumulating on Ozonium stages of Basidiomycetes. Numerous germination cultures have been made with ma- terial from two weeks to three months old. While this has af- forded some interesting suggestions, germination in any par- ticular medium has been, on the whole, erratic. The data are .4. Phymatotrichum omnivorum: acicular hyphae exhibiting charac- teristic modes of branching. reserved for a later report. The cultures which have been prepared from the newly infected root, as also those from erratic spore germination, have yielded a sterile mycelium which, while in itself distinctive, resembles only in a general way the mycelium found on the roots and in the soil. The mycelium in culture is hyaline, forming on young bean stems and on various other culture media a dense, slow-growing mat, seldom rising more than 3 mm. above the substratum, and never becoming fluffy in appearance. After standing for some weeks this mycelium becomes somewhat colored, as- suming a warm buff to light ochraceous buff. In culture the 1916] DUGGAR—THE TEXAS ROOT ROT FUNGUS GA? hyphae are likewise most diverse in diameter, varying from those 15-20» to others extremely delicate and flexuous (fig. 5). No truly acicular branches, however, are produced un- der ordinary cultural conditions. With age, the mycelium somewhat collapses toward the substratum and has a greater tendency to grow along the glass tube in the form of false strands. Grown in soil, by covering a vigorous growth on bean stems with a layer of loam, hyphae similar to those just des- eribed are produced; but, in addition, there are formed here and there vesicular enlargements, and the latter are sometimes реа" 9. w deaet omnivorum: hyphae grown on bean stems, from a D 60 days in clusters, the branches becoming two to three times forked. The conditions for conidial formation have not been deter- mined. It will be seen that the connection of the conidial stage with the Ozonium rests at present upon two classes of observation: (1) the presence in the conidial layer of hyphae and strands (bearing conidia) found to be identical with the character- istic mycelium on the roots of affected plants, and (2) the identity in artificial culture of the mycelium originating, on the one hand, from diseased roots, and, on the other, from the germination of the conidia. То complete the proof it would, of course, be necessary to secure positive results by inoculation with conidia, or better, positive results with a pure culture originating from conidia. Unfortunately this phase of the work has not been successfully developed. In ANGTON UN; € =з П MEn ASOC ara QO Se -m ` e i ech рди”. [ Vor. 3 22 ANNALS OF THE MISSOURI BOTANICAL GARDEN this connection it should be said that no inoculations carried out in the greenhouse up to the present have given positive results. As a source of infection I have employed (1) dis- eased cotton roots fresh from the field (showing the Ozoniwm in abundance), (2) fresh conidia, and (3) cultures from dis- eased roots. It is apparent that the conditions for infection have not been made satisfactory. Such experiments are to be continued both in the greenhouse and in the field. It has been found difficult to place the fungus satisfactorily in any established genus of the Hyphomycetes. While in the manner of conidial production it is undoubtedly related to such genera as Phymatotrichum, Botryosporium, Rhinotri- chum, ete., it does not exhibit all the characteristics of any of these genera. Nevertheless, it has seemed best, after ex- amining all available exsiccati material of forms which might be related, to place the fungus tentatively in the genus Phy- matotrichum, and, if Bonorden’s figure (Handb. d. allgem. Myk. pl. 8, f. 181) is correct, fairly close to P. pyramidale Bon. The fungus is clearly excluded from Botryosporium, the conidiophores of which are erect, with conidia produced on sterigmata. Slightly emended, the genus Phymatotr- chum would be of taxonomic convenience. In placing the Texas cotton fungus in this genus, I would not convey the impression that this fungus is considered to belong to the Ascomycetes. Accepting Shear’s specific name, a revised description of the organism is appended. Phymatotrichum omnivorum (Shear) Dugear, n. comb. Hyphae diverse, forming on the host (1) a loose weft of large, branched cells, producing more rigid hyphae with acic- 'It should be noted that the genus Phymatotrichum ër E d. allgem. Myk. p. 116. pl. 8, f. 181. n was at first reduced t seetion of Botrytis by Saceardo (Sylloge ere 86). | Later, however, he ion it to generic rank (Sylloge 16:1033. 02) P^ aecommodate a speeies of Oudemans. Costantin y Mucédinées San pp. 44-46. f. 12. 1888) Se a detailed de- scription a fungus, which was obv Ж considered We intei v pyramidale Bon., see the name Botryosporium pyramidale Cost ere 57 little doubt that the D figured by Cos — is “prope rly plac ed. However, the source of Costantin rial was apparently the original specimen of Bor onorden, and since his eg D in many Se from that of Bonorden, it is perhaps E to Ces estion the identi EA D the two fungi. Lindau (Rabenhorst's Kryptogam flora 1 (Abt. 8) :117 04) spe to ge the views of Costantin. He also vites As exsiccati, 2 Micr. sel. 1907. 1916] DUGGAR—THE TEXAS ROOT ROT FUNGUS 23 ular branches, these last often arising at right angles and op- posite, and (2) plectenchymatic strands; almost hyaline when young to cinnamon-brown in mature strands. Fertile hyphae arising irregularly from the large-celled mycelium or direct from cells of the strands, assurgent, simple or forked, with spore-bearing portion vesicular (spheroidal to ellipsoidal), often 20-28» in length and 15-20, in diameter. Spores finally arising also from undifferentiated hyphal and strand cells, hyaline, spheroidal to ovoidal, the spheroidal averaging 4.85.5 in diameter, the ovoidal measuring 5-6x(6-84; ex- treme diameters, 3.2 and 9.8 и. The conidial stage forms a con- tinuous pulverulent, sometimes crust-like, area on the soil. Hab. Hyphae on living roots of many plants and in soil, conidial stage on soil in the vicinity of diseased plants. Specimens have been deposited in the herbaria of the Mis- souri Botanical Garden and the Bureau of Plant Industry, Washington, and in the collection of Professor W. G. Farlow, Cambridge. BIBLIOGRAPHY Atkinson, G. F. (’93). Method for ке pure cultures of Pammel’s fungus of Texas root rot of cotton. Bot. Gaz. 18:16-19. 1893. Curtis, б. W. (92). Alfalfa root rot. Тех. Agr. Exp. Sta., Bul. 22:209-215. 1892. Duggar, B. M. (709). Fungous diseases of plants. pp. 479-481. f. 240. 1909. Heald, F. D. (709). Symptoms of disease in plants. Univ. Tex. Bul. 135:pp. 57-58. f. 56. 1909. , (11). Texas root rot of cotton. Tex. Dept. Agr., Bul. 22:303-308. 1911. , and ЗАН; Е. А. (711). List of hae” bgp and fungi occurring in Texas. x. Acad. Sei. s Trans, 11:1 ,—— s (12). A plant disease survey in the vieinity of San Antonio, exas. U.S. Dept. Agr., Bur. Pl. Ind., Bul. 226:pp. 26, 38, 41, 48, 49, 55, 57, „р 1912 Orton, W. A. (’02-’07). Plant diseases in the United States. U.S. t. Agr., Yearbook 1902:718; 1903:555; 1905:603, 604, 610; 1906: 506; 1007: 586. agre 02-185 D 8). Root rot of po or ''eotton blight ? Tex. Agr. Exp. „ Rept. 1:50-65. 1888; Ibid., Bul 4:18-33. 1888. -------, (789). Cottonroot-rot. Ibid., Rept. 2:61-86. pl. 1-5. 1889; Ibid., Bul. 7:1-30. pl. 1-5. 1889. Shear, C. L. (707). New species of fungi. Bul. Torr. Bot. Club 34:pp. 305-306. 1907. —__——., and Miles, С. Е. ('07). The control of pu root-rot of cotton. U: 8. Dept. Agr., Bur. РІ. Ind, Bul, 102:39-42. f. 3. 1907. -------, (7073). Texas root-rot of cotton. Ibid, Cire. 9:1-7. f. 1 CABBAGE YELLOWS AND THE RELATION Ob TEMPERATURE TO ITS OCCURRENCE JOSEPH C. GILMAN Formerly Rufus J. Lackland Fellow in the Henry Shaw School of Botany of ‘ashington University [INTRODUCTION In recent years the diseases of plants caused by fungi be- longing to the genus Fusarium have assumed greater and greater importance from an economic standpoint. A large amount of work has been done on the descriptions of such dis- eases on new hosts, and on the taxonomy of the genus Fusa- rium, but there has been comparatively little study of the re- lations of these fungi to their hosts, especially of the condi- tions under which members of this genus may become harm- ful parasites. Therefore, any work which throws light on this point is of value scientifically, first, because the mode of attack and the other relations of the parasitic species of Fusa- rium are all very closely related and very similar in their nature, and second, because of the possibility of throwing light on the problem of immunity or resistance of plants to the at- tack of parasitic organisms. The latter point is of particular interest, since it will be recalled that, up to the present time, practically the only control of the diseases caused by fungi belonging to this genus, has been by the selection or develop- ment of strains of the host resistant to fungous attack. While assisting in the work of the development of strains of cabbage resistant to yellows in Wisconsin, investigations were undertaken to find the cause of the disease and the rela- tions between host and parasite. During these investigations the relation of temperature to the occurrence of this disease was found to be of utmost importance, and the principal part of the work was accordingly devoted to this side of the prob- lem. Nevertheless, before taking up these observations and experiments in detail, the results of the investigations into the etiology and pathological anatomy of the disease should ANN. Mo. Bor. Garb., Vor. 3, 1916 (25) [Vor. 3 26 ANNALS OF THE MISSOURI BOTANICAL GARDEN be discussed, in order that the physiological relations between host and parasite may be understood more clearly. А brief resumé of the literature on cabbage yellows will show the state of our knowledge of this disease at the time these inves- tigations were taken up. History oF THE DISEASE The disease was first reported by Smith (799, 7997), ав occur- ring in New York State in 1895. He found the trouble ex- ceedingly severe, threatening ‘‘to put an end to the success- ful growing of eabbages in considerable districts.”’ He con- sidered that the disease was ‘‘due to a soil Fusarium” but made no inoeulation experiments. Aside from this observa- tion his only contribution to our knowledge of the disease was in relation to its persistence in the soil; the organism resisted drying in the laboratory for three and one-half years. Woods (799) showed that the characteristic symptom, the yellowing, was due to the presence of an increased amount of an oxidiz- ing enzyme, peroxidase, in the diseased leaf tissue. Norton and Symons (207) reported the presence of Ше disease in Maryland, but performed no experimental work. Harter (709), of the Bureau of Plant Industry, made inocu- lations of sterile soil with pure cultures of a Fusarium isolated from the stems of diseased cabbage plants. He was able to produce the characteristic symptoms in plants grown in that soil. In one trial, 83 per cent of these inoculations were suc- cessful; in a second, he reported that a large percentage of the plants showed typical symptoms, but no exact figure was given. He also made the statement that the fungus was a vascular parasite and formed microconidia in the vessels of the living plant. In addition to this paper Harter (712) has pub- lished merely a popular aceount of the disease. Manns (711) reported the disease as prevalent and destructive in Ohio but limited his work to field observations of a general nature. Jones (713, 714, 714%) in a series of papers reported the de- velopment of strains of winter and "kraut" types of eab- bage which are highly resistant to the attaek of this disease. These strains were developed by means of selection of sound 1916] GILMAN—CABBAGE YELLOWS 27 plants from badly diseased fields. In his last paper he re- ported that in the resistant strain 100 per cent formed com- mercial heads, or a yield of 18.8 tons to the acre; on the other hand, in the commercial strain used as controls, only 46 per cent of the plants lived and 24.2 per cent headed, or a yield of 2 tons to the acre. These results show that, as far as practice is concerned, the disease has been controlled, but much remains to be done on the other aspects of the problem of the relation of host and parasite. Before discussing these phases, however, a brief description of the disease will not be out of place. SYMPTOMS OF THE DISEASE The first evidence of the disease in the greenhouse is found on very young seedlings, often just after the appearance of the first true leaf, and is characterized by a rapid wilting of the cotyledons and dying of the roots while the stem is still turgid and, to all external appearances, normal. If, however, the conditions are not favorable for the attack of the fungus so early in the life of the host, the characteristic symptom —the yellowing of the leaf, to which the disease owes the name of ‘‘yellows’’—is found. This yellowing may invade the en- tire plant, in which case wilting and death rapidly follow, or it may be confined to merely one side of the plant or leaf. If this one-sided invasion occurs, the plant or leaf ceases growth on the diseased side, but the green portions continue their development, bringing about a curvature of the plant or leaf toward the diseased area. This type is most fre- quently found on plants grown in the seed-beds in infected soil. These one-sided plants are usually stunted with the leaves loosely attached to the stem, so that they fall at the touch. If transplanted into the field such plants may die immediately, or if conditions are favorable for their development, they may live all summer, becoming stunted individuals with the lower leaves dying and dropping off, leaving a tuft of living leaves at the tip of a bare stalk. When healthy seedlings are transplanted to diseased soil the same characteristics occur ; some plants die immediately—first, however, losing their chlo- [Vor. 3 28 ANNALS OF THE MISSOURI BOTANICAL GARDEN rophyll—while others become one-sided and stunted, but live throughout the summer. "The latter rarely form heads. CAUSAL ORGANISM TAXONOMY Cabbage yellows is caused by a soil fungus belonging to the genus Fusarium. The organism was first described by Wollenweber (713) who, basing his classification on the work of Appel and Wollenweber (710), placed it in the section Ele- gans and named it Fusarium conglutinans Wollenw. The de- seription given by this author is as follows: s EE n. sp. differs from F. orthoceras he absence of a wine-red color on rice which is a "striking dier of typical species of the section Elegans. Vascular parasite, cause of wilt disease of Brassica KE var. capitata (proved by Erwin F. acte L. R. Jones, L. L. Harter) in the United States of Americ This deserip- tion was based on a culture from the Laboratory of Plant Pathol- ogy of the Uni- versity of Wis- consin. The same е year Stevens (218) ascribed the yellows to Fusa- rium Brassicae Fig. Conidia е of vi ae % Thum., citing eet s fluid after 48 hou a, Culture V; b, A ture II; c, Culture I. Gei унда ат x ge Harter ( 09) as his authority in spite of the following facts: first, that Harter specifically stated that he was working with an undescribed species and, second, that Wollenweber had included Harter’s organism in his new species, F. conglutinans. Moreover, the organism that is parasitic on cabbage in the United States differs from Fu- sarium Brassicae Thüm. as described by De Thümen (280) 1916] GILMAN—CABBAGE YELLOWS 29 in the following respects: Fusarium Brassicae forms sporo- dochia, while in F. conglutinans these are much reduced and usually not formed at all. F. Brassicae has conidia which are two-septate, while F. conglutinans has conidia, the majority of which are non-septate with a few one- and three-septate forms, two-septate spores not appearing. The main point of resemblance, from the description of F. Brassicae, is that the spore measurements fall within the same limits—a fact which, in view of the above differences, would scarcely suffice to put the two as synonymous. Fig. 2. Conidia production of F. conglutinans in Uschinsky's fluid after 48 hours. Culture I. Camera lueida sketeh >x 1000. While the above description by Wollenweber is perhaps sufficient to differentiate F. conglutinans as a distinct form, it is hardly adequate as a diagnosis of the species; moreover, the question may be raised as to whether a physiological char- acter, such as color production on a special medium, which has not been regarded as of specific rank in related genera, is sufficient basis for the establishment of a new species in the genus Fusarium. This, of course, introduces a new factor into the taxonomy of this genus, but the writer would hold it [Vor. 3 30 ANNALS OF THE MISSOURI BOTANICAL GARDEN Fig. 3. Production of era? Ken of F. conglutinans in Usehin s fluid after five days. Culture xL рн lucida sketch x 1000. to be justifiable in such a group as the genus Fusarium, where a classification based on morphology alone has led and would continue to lead to confusion in many cases. That this character is con- stant in the case of F. con- glutinans, there can be no doubt, but as additional evi- dence, some forty-three cul- tures of this organism have been maintained in the lab- oratory in connection with this work for a period of from six months to two years, and in no case did they produce red color on rice media, while cultures of F. orthoceras, carried as controls, did. Moreover, on other media the organism maintained constant similar- ities with cultures from similar sources which oninoculation into the host produced the disease. The organisms were grown on potato hard agar, dextrose bouillon agar, soil extract agar, cooked potato plugs, cooked potato stems, and cooked rice. The mycelium in all cases grew well, giving a white fluffy growth at first, which gradu- ally turned cream color, and in old cultures showed ochreous to brown strands in the aerial mycelium in the upper part of the tube. Spores of the “тпісто” type were found in all cultures in great abundance, espec- tally during the early part of the growth of the cultures. The production of aérial mycelium was most abundant in those cases where the amount of carbohydrate in Fig. 4. Method of branching of myce li о Camera lucida es x 1000. the substratum was greatest or most available. 1916] GILMAN—CABBAGE YELLOWS 31 After growth of a few weeks chlamydospores were found in most of the cultures, the microspores were beginning to be- come abnormal, and the few macrospores were also breaking down. The macrospores were found to be produced best on potato stems, next best on the potato agar, while very few ap- peared on cooked potato plugs and cooked rice. Besides the tube cultures, hang- ing-drop cultures in a modified Uschinsky’s fluid! were observed. In this medium the fungus pro- се duced microspores abundantly in cultures only two days old when 2 kept ай room temperatures. Chlamydospores were found to begin to form in cultures but five days old, although they did not mature insoshorta time. Usually Ж the first chlamydospores occurred terminally; later other parts of the mycelium rounded up to form the intercalary spores. A revised deseription of the fungus is as follows: Fusarium conglutinans Wollenw. Sporodochia laeking or greatly А Fig. 5. Conidia of Е. conglu- reduced; pionnotes never present. tinavs. Culture LVI on potato idi idi m. ida sketeh x Conidia borne on short conidi- gio" Camera luci ophores strewn throughout the mycelium, majority non-septate, a few one-septate and three-septate. The non-septate conidia ovoid to ellipsoidal, hyaline, 2.5-4 X 6-154, the majority being 2.5-3 X 7-104. One-septate conidia hyaline, cylindrical, with This medium was га modification of the standard Uschinsky’s fluid, and was made up as follow E она ЗБРОЯ 1000 grams Magnesium sulphate ..... 0.3 grams a | EE EE 30 grams CES phosphate .. 2 grams oe chloride `. 5 grams Ammonium tartrate ..... 6 grams Caleium AA ua ces 0.1 grams od аа, «a. В grams gr The solution was sterilized at ten pounds pressure for twenty minutes in the autoclave. [Vor. 3 32 ANNALS OF THE MISSOURI BOTANICAL GARDEN rounded ends, long axis slightly bent, dimensions 4 X 19u. Three-septate conidia fusiform, hyaline, with both end-cells tapering and with rounded tips, no sharply differentiated foot, dimensions 3.5-5.5 X 25-334. Conidia with higher septation very rare. In culture aérial mycelium white at first, becoming cream- colored and finally showing a development of ochreous strands of thallo-plectenchymatic tissue throughout, but no sclerotia. Grows well on potato agar, dextrose bouillon agar, Uschinsky’s fluid, cooked rice, cooked potato plugs, and potato stems. On no medium is there any color production except the slight yellowing spoken of above. In older cultures terminal or in- tercalary chlamydospores are produced. They are usually one-, sometimes two-celled, spherical to ovoid with a thick irregular wall, frequently slightly colored. Dimensions 7-12 X 7-15». The fungus is found in the soil and is a vascular parasite attacking cabbage, Brassica oleracea var. capitata, causing the yellows, or wilt disease. It has also been isolated as a sapro- phyte from China aster and tubers of potato (Lewis, '13).' TEMPERATURE STUDIES Tn view of the later work in regard to the relation of temper- ature to the occurrence of the disease, it will be well to dis- cuss briefly this relation for the fungus in pure culture, both as to growth and germination of conidia. The latter will be considered first. In order to obtain spores of the fungus free from pieces of mycelium, a bit of mycelium was placed in a hanging-drop of Uschinsky’s fluid in a Van Tieghem cell which was partially filled with Uschinsky’s fluid below. The spores were formed abundantly at room temperatures in forty-eight hours and al- lowed to drop into the lower liquid from which they were 1 Through the kindness of Dr. W. J. Morse, of the Maine Agricultural Expe- ent Station, ior ore rs of these two strains of Fusarium conglutinans Wes? obtained, and in ations made on February 2, 1915, on five plants each, again on Ke 26, 1915, with а strain from aster on ten plants; both gave negative results. This would indicate Ser they Geste to a saprophytic strain, although the results might be due to the fact that the — ad bee so long in eulture that they ha d lost their гр or to the small number of trials made. Control cultures of this fungus from ca abbage, however gave 80 per cent infeetion in the first case and 100 per cent in the second tria 1916] GILMAN——CABBAGE YELLOWS 20 transferred to other Van Tieghem cells by means of a sterile pipette. They were immediately placed in the incubators at the desired temperatures, and observed at intervals for ger- mination. All observations were made in duplicate, and the trials were repeated twice to verify them. Тһе temperatures used were 8-10°C., 10-12°C., 16°C., 21°C., and 33°C. The results are brought together in table 1. TABLE I GERMINATION OF CONIDIA OF FUSARIUM Aerer ново AT VARIOUS TEMPERATURE Germination (+) at the following temperatures Hours of 8-10? C. 10-12? C. 1626. 212; е С. exposure Trial no. Trial no. Trial no. Trial no. Trial no. 1 2 1 2 1 2 1 2 1 2 I ee — — БЕН IH == = = = =з а Да Жы. -- — -= E ща PS Es E: zz - Hi Mor — — — == ши = = aia 4 4 Dro АГ — — — — — = 7 ВЕЕ + + NEC — — — 7 +* T* aR E + + Lae -- -- -- — + + + + + + BE... = — — — + + + + + “Б Ee — -- — | —t + + + 4- ch mE ШЕ». — — — 4 + + A 4- + ater test in which more frequent observations were made, conidia at this Senaye (16°C.) germinated 12 hours after the beginning of the exposure. ł At this temperature (10-12° C.) spores were found to germinate after 36 n a later experiment. The number that germinated, however, was very small and the growth eec Zi slow It will be noted that, as was to be expected, spores of the fungus germinated best at the higher temperatures of the experiment, although they were able to grow slowly at the lower temperatures. These facts are further borne out by the growth of the fungus on potato agar. Transfers of a bit of the mycelium from a rapidly growing culture were placed in the center of plates of potato hard agar, and the plates were then placed in the incubators at the desired temperatures. 3 [VoL. 5 34 ANNALS OF THE MISSOURI BOTANICAL GARDEN Three plates were carried at each temperature, and measure- ments of the growth of the colonies of the mycelium were made each day for ten days, after which time the experiment was dis- continued because of the contamination of some of the plates and the drying out of others. The results are given in ta- e II. TABLE II GROWTH OF FUSARIUM CONGLUTINANS AT VARIOUS TEMPERATURES Diameter of colony in cm. at various temperatures Age of 4-8? C. 18? C. 21-22? C. 25 C. colony in days Plate no. Plate no. Plate no. Plate no. EI3418111342151]114213 11-21 3 ESO Lore» ERREUR EE а EH ETS Ы а О. О.О ТТВ E E EE ЖЕЕ asal welt 9:4 0.1 0.H 1.8 1,8 12 1.29 208 11 assise EVI 0.1] 0.1| 0.1} 0.6| 0.6| 0.6| 2.0| 2.01 1.8| 2.0| 2.2| 1.7 ОТТ РЕТОТИТТРИРЕ 0.21 0.2 0. 11.00.9 0.9) 3.0] 2.8] 2.4] 2.9] 3.2] 2.9 Boule ere ала 0.4) 0.5 0.4! 1.2) 1.1] 1.2) 3.6] 3.4] 3.0] 3.4, —*| 3.1 Dich Ries PROP 0.5} 0.7 0.5 1.41 1.2) 1.3] 4.0] 3.8] 3.4] 3.8 3.7 ОРУУ КЕРЕЙ 0.7) 0.8| 0.9) 1.5) 1.3] 1.51 4543 —*| 4.6 4.4 ТТК ТТТ KH 111: 1.8] 1.7] 1.7] 5.1] 4.8 5.1]. 5.3 SEN 1.4| 1.5| 1.3] 2.0] 1.8| 1.8 5.5] 5.4]. 5.6. .0 j| BCEE 1.6| 1.7| 1.4| 2.2| 2.2| 1.9| 6.2| 6.0}. —* 6.6 gieler? growth Pay S. 0.160.170.140.220.220.190.620.600.560 6210.50|0.66. Average for each OI крал 0.16 0.21 0.59 0.69 *Contaminated. If the growth of Fusarium conglutinans be compared with that of some of our more common saprophytic forms as, for example, Penicillium glaucum, or Aspergillus niger, as re- ported in the literature, it will be noted that, while the op- timum of these forms is also high, they can grow better than Е. conglutinans at the lower temperatures. In other respects the eurves of growth of these forms would approximate one- another very closely. 1916] GILMAN—CABBAGE YELLOWS 35 No attempt was made to find the maximum and minimum growth temperature for this fungus, because the object of the work was to find, if possible, an explanation for the fact that yellows occurred in the host at high temperatures rather than at low. This relation will be discussed later when a full review of the points involved will be taken up. INOCULATION EXPERIMENTS The first inoculation experiments were tried during the sum- mer of 1912. On July 17 five flats of soil were planted to cabbage; three contained soil brought from the experimental plot at Racine and two, normal greenhouse soil. One of the latter was left as a control, and the other was inoculated with spores from a pure culture of the fungus. These flats were kept shaded on the north side of some shrubbery in the path- ological garden and no typical yellows had appeared by Sep- tember 7, when they were discarded. On August 23 five plants in the pathological garden were inoculated by placing myce- lium of a rapidly growing culture in contact with the roots. No disease was found up to October 22, when frost killed the plants. The seedlings were two weeks old at the time of inoc- ulation. Again, on September 10, thirty healthy plants were transferred to three flats of soil brought from the experi- mental plots at Racine, but no disease was found in any of the flats by December 2. On January 6, 1913, twelve pots of sterile soil were planted to cabbage and inoculated by stirring cultures of Fusarium into the pots. Twelve pots of diseased soil from the experi- mental plot, eight of sterilized soil, and four of normal undis- eased soil were planted as controls. Spores were abundant in all the cultures used. No yellows had appeared by April 29, and the plants were then pulled and the pots replanted. Instead of keeping this second lot in the open greenhouse, [ VoL. 3 36 ANNALS OF THE MISSOURI BOTANICAL GARDEN however, they were placed under a glass, such as is used in a forcing-bed, thereby giving a higher temperature than could be attained in the open house. On January 14 the plants in one pot of inoculated soil showed the characteristic symp- toms, and on July 10 the plants in a second pot had suc- cumbed. On July 12 a third pot contained plants showing the disease. Damping off due to Rhizoctonia interfered with the value of this trial. Тһе diseased plants were plated out on potato hard agar in all cases, and the typical Fusarium found to be present. As will be noted, it was only after the tem- perature of the pots had been raised that there was any ос- currence of the disease. Curves showing the temperature attained by placing the plants under the glass are given in fig. 6 Ma 4. ae з гот " ши ТТ ТЕБЕН a А CEO BRSRRRS ЕЕЕ ie ake и wi s ming; Li ининин най E m a ak Li Y таяз итиини и! 11 * МЕНН ШЕН a г. am aE EST SE CTIE? ] ай coos 5 tN | uu HHTH Wei П BEENS. 2i LEIT Li Li ІП ALI Li + L: LLLELLET завшиша! I1 Li Іп ПІ! Li Ka Li а жи ята и: Oh е! 14 LI LEI ІТ! Li 9; Li TILLLLEI LELLLILII li 4 Lii Lil Li Li LELLLLEI TELI ial Li Іп LK LI e u =" ВАН = EECH здү шин ТИ ГІ CHAT En L Sp H М = +44 9; Т EI) HI о Li пеи р IIT II LLLLLIKI CEEE Li L1 LILLLLEL ELL? 11 шишиши 1411 {1 Ew LI вишна! EET EI li ELLE) LLLI Ы © LI CSL «куш, LI LLLLLLEI LETE L: Y Tt massa. LLLLILLILLI LI п LLI Li су Li шааж а: ТТ LI LLLLLLT I LILI Li b: Fig. 6. iagram showing temperature under glass in кы. ina pod inoculation experiments, Мау 5- 1914. On May 10, 1913, another inoculation experiment was started with seedlings grown in normal greenhouse soil. The plants used were about ten days old — just showing the first true leaf. Five pots of soil were used, five seedlings in each pot. Pot 1 was left as a control, while the roots of the plants in the other pots were dipped into a suspension of spores from a pure culture and immediately planted. On May 13 they had all recovered from the effects of transplanting and were in good 1916] 1 GILMAN—CABBAGE YELLOWS Bt condition. On July 10 symptoms of yellows showed in pots 4 and 5; on July 14 the plants in pot 3 became diseased, but no disease was found in pots 1 or 2. The plants were plated on potato hard agar, and F. conglutinans was recovered from plants from pots 3, 4, and 5. None was found on seedlings plated from pots 1 and 2. Culture V, which upon reinocula- tion again produced the disease, was one of these cultures (table v). On May 27, 1913, a more extensive inoculation experiment was started; part of the pots containing the plants were placed in the greenhouse and part in the pathologieal garden. As the greenhouse was not heated, the difference of conditions was not noticeable, and, therefore, the results are combined. For this trial twenty pots were planted to cabbage. These pots were in duplieate, ten being placed in the greenhouse and ten in the soil of the garden. The treatment of the soil in these pots was as follows: Eight pots contained soil from the ex- perimental plots at Raeine, and eight others normal green- house soil These sixteen containers were sterilized in an autoclave at eleven pounds pressure for four hours. Four of each of them were used as controls, four were inoculated with pure eultures of F. conglutinans, and the remaining four were inoculated with wilted leaves of the diseased plants. Two pots of normal greenhouse soil and two of normal diseased soil were added to the series as controls. The cultures were added by mixing them with the surface soil in the pots. The seed used were not good, and, therefore, the plants did not eome up well, so that in the following pots there were no plants: one pot of diseased soil sterilized but not inoculated ; two pots of diseased soil sterilized and inoeulated with pure cultures; two of diseased soil sterilized and inoculated with the leaves; one pot of normal greenhouse soil not inoculated ; and one of sterilized greenhouse soil inoculated with leaves. The results of this experiment are given in table rrr. [VoL. 5 38 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE III RESULTS OF INOCULATIONS WITH PURE CULTURES OF FUSARIUM CONGLUTINANS Plants per pot Pot no, Kind of soil Treatment of soil Percent- Total No. age no. diseased | diseased 1 | Infected.......... беегіпгей................. 7 0 0 2 | Infected........... Sterilized oii Ve RN oo be Ü. ondas d 3 | Infected ter'z'd and inoc'l'd........ 4 3 75 4 | Infected........... Ster'z'd and inoc’l’d........ 1 1 100 5 | Infected........... Монша tics cai Lun 4 80 6 | Infected........... Normál i. Ae Bee aer carne 6 6 100 7 | Uninfected........ МОКа beck eius 0 0 8 | Uninfected........ Мота ooh vvv 1 PEREP ETT 9 | Uninfected........ Ster'z'd and inoc’l’d........ 3 1 33 10 | Uninfected. ....... Ster'z'd and inoc'l'd........ 5 5 100 11 | Uninfected........ Ster'z'd and inoc'l'd........ 1 1 100 12 | Uninfected........ Ster'z'd and inoc'l'd........ u 1 50 13 | Uninfected........ Sterilized................. 6 0 0 14 | Uninfected........ Sterilized. оо: 4 0 0 15 | Uninfected........ Ster'z'd and infect. leaves... 2 1 50 16 | Uninfected........ Ster'z'd and infect. leaves... O VT ays Ee The results are not as conclusive as they might be, how- ever, as the number of plants used was very small due to the poor seed mentioned above. The pots in which no plants grew in either of the duplicates were omitted. An idea of the appearance of the plants at the time of making counts may be had from pl. 1, fig. 2. It should be noted that these successful inoculations were made at the warmest time in the summer. Plates were made on July 14 from plants in each pot, and in all cases the diseased seedlings gave pure cultures of F. conglutinans, while those from the controls remained sterile. Culture X was one of these and upon reinoculation again produced the disease (table v). On June 12 the above experiment was repeated, using three flats of soil (from the experimental field), two of which had been sterilized at eleven pounds pressure for four hours on two successive days, the other remaining untreated. One of 1916] GILMAN—CABBAGE YELLOWS 39 the sterile flats was inoculated by stirring into its surface ten pure cultures of F. conglutinans which were sporulating abundantly. Then all the flats were planted. As an addi- tional control, a flat of normal greenhouse soil was placed in the series. On July 7 yellows began to appear in the steril- ized inoculated flat and continued to spread until July 16, when the experiment was concluded by making plates from the plants from the sterilized inoculated, and the sterilized flats. F. conglutinans was found as the cause of the yellowing in all the plants, while the control plants remained sterile. This is shown well in pl. 2, figs. 15 and 16. Exact counts were not made. Again, on June 29, four pots of sterile greenhouse soil were inoculated with cultures of F. conglutinans, and on July 24 yellows was found in all four of the pots. On July 11 inocu- lations of individual plants were repeated by dipping wounded plant roots into suspensions of the spores of the fun- gus in sterile water. Adequate controls were included in this series and in all cases the control plants remained healthy, while among the inoculated plants, 50 per cent of the indi- viduals showed the characteristic symptoms on July 24, when the experiment was discontinued. VIRULENCE OF CULTURES Pure cultures of F. conglutinans vary greatly in their viru- lence, and the cause of this variation is not certain. From inoculation experiments it would seem that, in general, the longer the organism has been carried in culture the greater is the probability that it has lost its virulence. On the other hand, drying in culture seems to have little or no ill effect on the virulence of the organism. The susceptibility of the host must also be considered as an important factor when the fungus-host relation hangs in such a delicate balance, and the source of the culture is always [Vor. 3 40 ANNALS OF THE MISSOURI BOTANICAL GARDEN of importance also. The medium upon which the culture is grown and the state of the mycelium and spores have been pointed out by Wollenweber as important factors in other species of Fusarium which produce plant disease, and doubt- less they bear their part in the irregularity of the results presented here. In a series of inoculation experiments made at the Missouri Botanical Garden, recently isolated cultures were used as sources of infection. The cultures were grown on cooked po- tato stems, and inoculation was effected by placing a bit of the culture tissue in contact with a wound on the hypocotyl of the plant. The plants were in the cotyledonous stage, and after inoculation were placed in normal uninfected greenhouse soil. Five seedlings were placed in each pot. Table rv gives the results of the experiments. TABLE IV PRELIMINARY STUDY OF VARIATION OF VIRULENCE OF FUSARIUM CONGLUTINANS IN PURE CULTURE No. of Per cent of Culture Total no. diseased diseased number isolation of plants BN E 7/14/13 40 30 75.0 те 11/17/14 15 13 86.6 он. 2/1/15 5 5 100.0 EE РУТИНЫ И 10 0. It will be noted that where the larger number of plants was used the percentage of infection fell. This result might have been expected if the age of the cultures used and the great variation in susceptibility of the host plant were taken into consideration, but to gather more data on these points a trial was made with a large series of cultures that had been isolated at various times and also from various sources. Table v gives the data on the inoculation experiment which was carried out similarly to the one just reported. 1916] GILMAN—CABBAGE YELLOWS RESULT OF INOCULATION VIRULENCE OF FUSARIU 41 TABLE V EXPERIMENT SHOWING VARIATIONS IN M CONGLUTINANS IN PURE CULTURE Culture Species number [ce F. conglutinans.. || eem F. conglutinans.. ` без rs conglutinans E F. conglutinans 1 А conglutinans лл F. conglutinans BI F. conglutina EI . z: F. conglutinans NIV F. conglutinans ARV tes F. conglutinans INVI. F. conglutinans XVIII Undetermined.. 295b M conglutinans КИТ Е. conglutinans 9,4 m M F. conglutinans XXII....|F. conglutinans XXIII. F. conglutinans XXIV. F. conglutinans XXV.. onglutinans XXVI. F. orthoceras XXVII Undetermined XXVIII nglutinans XXIX. Undetermined Э.Ө. conglutinans XXXII...) F. conglutinans.. XXXIII..|Undetermine XXXV...|F. conglutinans.. XXXVI. ДЕ. conglutinans.. XXXVII .|F. conglutinans.. XXXVIII|F. conglutinans.. XXXIX. .|F. conglutinans.. П . conglutinans.. #@ tee F. conglutinans.. XLIII....|Undetermined.... XLIV. F. conglutinans ESI os F. conglutinans ALVE, F. congluiinans XLVII...|F. conglutinans SE? F. conglutinans га | OPE conglutinans ЗА 2 .... F. conglutin LIV. Undetermined Шу F. conglutinans БТА d . conglutinans Control. ..|No fungus....... Source Ca Jd Ca Ca bbage.... ст за а `a ОооОоОООООО р © & D Ca Cab H ET Er ES Em KEE a bbage.... uliflower. . a liflower. J age.. bbage. E uliflower. . bage... Pathogenicity Date of Damped| Үеі- Healthy isola- off lowed Hon р S LEE t cu i Per Per Per No.|cent| No.|cent| No. |cent 5/16/ ( 0 ( ( 5| 100 5/12/ ( 0 ( 5| 100 5/10/ 20 B] 6 te 20 5/10/ 20 ( 4| 80 /14/ ( ( 100 //14/ 2 2 60 г/14/ 10 ( 0 /14/ ( 2| 4 3| 60 г /14/ ( ( 5| 100 7/14/ ( 4 8 20 ech ( ( ( ( 00 7 /26/ ( 100 6/ ( 3| 6( 40 1/26/ ( 2| 4 60 г /26/ ( ( ( 100 //26/ ( ( ( 100 7/26/ ( ( ( 100 '/26/ ( ( ( 100 '/27/ 4| 8 2 ( 0 )/ 6/ ( ( ( ( 100 / 4/ ( ( ( ( 100 / 4/ ( ( ( 100 /24/ ( ( ( 100 6/28/ ( ər 46 40 6/28/ 2 4 8 ( ) /28/ OR e ( 3| 60 /24/ 20 ( 4| 80 6/24/ 2| 40] ( 3| € 6/28/] 2 2 3| 60 6/28/1 ( í 5| 1( 6/28/1 1 20| i 4| 80 1/ 3/14 5| 100 ( ) 2/14/14 2| 40 4( 1 20 cl oio e A 20 ( 4| 8 eue. ( 0 ( ( 5| 100 Cae 4 66 ( ( 2| 40 4/13/14 20 ( 4| 80 3/12/14 ( 0 & 6 40 5/ 6/14 ( 0 ( 5| 100 5/ 6/14 ( 0 2 4 3| 60 5/18/14 ( 0 a 4 š C 9/24/14 ( 0 ( 5| 10 11/17/14 0 0 4| 80 20 2/ 1/15| of o 5| 100 EN H 0 ( 0 t 90 *Isolated by Lewis ('13) at the Maine Agricultural Experiment Station and determined by H. W. Wollenweber, Bureau of Plant Industry, Washington, D. C. [ VoL. 3 42 ANNALS OF THE MISSOURI BOTANICAL GARDEN The cultures were all prepared in the same way for the experiment. They were all grown on cooked potato stems, and were of the same age. Each culture was used to inoculate five plants by inserting a bit of the mycelium into the hypo- cotyl of young seedlings still in the cotyledonous stage. The previous history of the cultures, of course, differed for the individual. Cultures VI, XVII, and XIX had been allowed to dry out on potato hard agar for fourteen months, that is, from July 14, 1913, to September 26, 1914, and then were transferred to cooked potato stems. On January 12, 1915, they were again transferred to fresh cooked potato stems, and these cultures were used in the experiment. Al- though two of them (VI and XVII) apparently lost their virulenee, the third (XIX) retained its ability to attack the host even after this severe drying. Other strains which had not been allowed to dry out but which were kept on fresh media, possessed no greater virulence, nor did any greater percentage of them exhibit pathogenicity. The length of time the organism has been in culture seems to be a more important factor; for cultures isolated late in 1914 showed proportionally a larger number virulent than did those isolated at earlier dates. In addition, the more recent isolations showed the greater virulence. That this is not invariable, however, is shown by the fact that many of the cultures first isolated still retained their virulence, viz., ХХХ, XXXII, XXXVII, all three of which were isolated on June 28, 1912. The source of the culture seems to have greater influence. Of the six strains of F. conglutinans isolated from cauliflower grown in diseased soil and apparently attacked with yellows, but one (XX) showed any ability to infect cabbage and that only to a limited extent. Strains from aster and potato, kindly furnished by Dr. W. J. Morse of the Maine Agricultural Experiment Station, also gave negative results when inocu- lated into cabbage. F. orthoceras, which had been isolated from the stem of a diseased cabbage plant, was introduced into the series as a control. A number of undetermined Fu- sarium cultures which had been isolated from cabbage, cauli- ; 19161 GILMAN—CABBAGE YELLOWS 43 flower, and China aster were added for the same reason. None of these latter were capable of infecting the living cab- bage plant. SUSCEPTIBILITY OF HOST That the susceptibility of the host plant must also play an important part in this question of inoculation is shown by the fact that so few of the cultures gave a perfect (100 per cent) infection, although the inoculations were made with parts of the same culture on plants from the same pot and under as identical conditions as possible. Further evidence on this point was also shown when the difference in the length of the incubation period of any one culture was noted on plants of the same variety and age. For example, in the last experiment observations were made daily in the greenhouse, and the condition of the plants noted. The results of these observations are brought together for a few of the cultures in table vt. TABLE VI RESULTS OF OBSERVATIONS ON INDIVIDUAL SUSCEPTIBILITY AS SHOWN BY THE INCUBATION PERIOD UPON INOCULATION Number of diseased plants in each pot at the various days of incubation Culture no. 15 16 17 19 23 40 50 +. А отказ. 1 2 3 3 3 3 1 So evo ы Ыс» 2 2 2 3 3 3 | beo deus gen ЕРТ 2 3 3 3 3 dE EE ose i 1 1 4 4 4 5 ыс ли Beret BEBE ER 2 3 3 8 22,55 SS Ee aes. 1 2 3 4 Control... EEN ҒАС ХЕ А E ma SE са = Thus it is shown that not only were some virulent cultures slower in taking effect than others, but that the individual plants were markedly different in their ability to resist the fungus. Although what constitutes such resistance has not been worked out, a little evidence gathered during these in- vestigations may well be presented here. [Vor. 3 44 ANNALS OF THE MISSOURI BOTANICAL GARDEN In the field it was noted that the plants of the resistant strains of cabbage were, as a rule, larger than plants of the commercial strains of the same age. The first year it was thought that this difference in size might be due to crowding in the seed-bed of the plants of the commercial strain, chiefly beeause the amount of available seed of the resistant varie- ties was limited, while that of the commercial strain was plentiful. When this faet repeated it- self over three years, experiments in the laboratory were run 7. Fungous hyphae obtained by disseetion of to account for the diseased stem after boiling in KOH solution. Camera difference. Seeds lucida sketeh. were placed between moist filter paper in petri dishes and allowed to germinate. Twenty-five seeds were placed in each dish, and two dishes of each strain, VIII a-16 and commercial Danish Ball-head, were germinated. After wetting up the filter paper with distilled water the dishes were all placed in an incubator at 22°C. The seeds of the resistant strain germinated twelve hours before those of the commercial varieties, three days after the begin- ning of the experiment. Plate 2, figs. 5 and 6 give an idea of the appearance of the seedlings at this time, under similar conditions of moisture and temperature. This characteristic of growth suggested that there might be a considerable difference in osmotic pressure between the root cells of the two strains, and experimental work was un- dertaken to determine whether the threshold of plasmolysis of the two strands differed toward NaCl solution as a plas- molytie agent. Two trials were made using the root-hairs as indicators, but in neither case was any difference between the threshold of plasmolysis of the resistant strain and that of the susceptible strain found. 1916] GILMAN—CABBAGE YELLOWS 45 Hosr RELATIONS MORPHOLOGY The distribution of the fungus in the living host tissues is limited to the vascular bundles. This fact was first shown in making plates from old stems of diseased plants. The stems were cut cross-wise in thin sections and, after sterilization in hydrogen peroxide for five minutes and washing in sterile water, were laid on the surface of poured plates of potato hard agar. Invariably the first growth of the mycelium ap- peared from the fibro-vascular ring (pl. 2, fig. 14). Upon dis- section of diseased seedlings the hyphae were demonstrable ` Fig Cross-section of vascular bundle from diseased cabbage stem, showing distribution of fungus in vessels. Note preponderance of cut ends of hyphae. Stained with Pianeze IIIb. Camera lucida sketch x 1000. traversing the lumina of the bundles longitudinally. The stems were first boiled for five minutes in a 5 per cent potassium hydroxide solution and then dissected under a hand lens. The final examination was made under the compound micro- scope. In no case was a very large amount of mycelium found in any single vessel (fig. 7). In later work the diseased stems were imbedded in paraffin in the usual manner, after fixing in Gilson’s solution, and stained with Pianeze IIIb, as recommended by Vaughan (214). The fungus stained а deep red, while Ше host tissue was col- [Vor. 3 46 ANNALS OF THE MISSOURI BOTANICAL GARDEN ored green. Longitudinal sections showed the hyphae of the fungus running longitudinally in the lumina of the spiral vessels and the bast fibres. Cross-sections showed only the cut ends of the fungus. Drawings illustrating these facts are shown in figs. 8 and 9 which were made with the aid of a camera lucida. It was found that besides the purely vegetative hyphae, the fungus produced conidia in the vessels of the host (fig. 10). Those spores observed in the host tissue were all of the unicellular type. All the evidence shows that the fungus at- tacks the root first, but Just how remains to be worked out. After entering the host, it is con- fined to the vascular system. The fungus was never isolated from 42-2 the stem until gcn Ñ tudinal section of a marked yellowing in diseased of the leaves ap- Mr уа, peared, although it hae ZZ Was always present Camera lucida in the tissues before sketch X 1000. they had been killed. This fact is brought out in pl. 2, fig. 12 in which is shown a branched plant, one branch of which was at- tacked, while the other remained healthy in appearance. The leaf at E was still alive although one side of it was yellow. The stem of the plant appeared normal externally. The fungus, however, was isolated from the stem below the branching and Fig. 10. Longitudinal section of ALI points D qud Z pu thie Ме Geet ote инш, diseased branch, while the parts Stained with Pianeze IIIb. Camera at B and C on the other branch BEER remained sterile. The plate made from this plant is shown in pl. 2, fig. 13. 1916] GILMAN——CABBAGE YELLOWS 47 After the death of the host the fungus traverses all the tissues, sporulating at the surface and within the host also. In this way the fungus is able to return to the soil. Whether it may winter over in the host tissue was tested by marking plants which have been killed by the yellows in the field in —— (? 1913, and then bringing these plants into the laboratory in the spring of 1914. The stumps were first freed from the soil by brushing them under water and then wash- ing in running water for fifteen minutes. After this washing the stalks were di- vided into equal portions and placed = two flats of Sterile Fig. ll. Conidia obtained from over- greenhouse soil (sterilized ID wintered cabbage stem in spring of 1915. the autoclave at eleven Camera lucida sketch x 800. pounds pressure for five hours) and left for twenty-four days, after which time the flats were planted to cabbage on June 3, 1914. Yellows was found in both flats on J uly 6, 1914, show- ing that the fungus was able to get back into the soil from these stems, or that the roots coming in contact with the stems were attacked. That the fungus may live over in the soil was first shown by E. F. Smith (99, '99*), when he found that the organ- ism in the soil was able to withstand dry- ing in the laboratory for three and one-half <-> years. Dr. M. P. Henderson in some un- published studies on Phoma lingam showed ET ig E inadvertently that the stumps of cabbage tem of over-win- are not necessary for the transmission of tucida sketch, Сашега Fusarium conglutinans, as it is able to live in the soil. He found that F. conglutinans was still present and virulent in soil that had been sifted through a fine sieve. [ VoL. 5 48 ANNALS OF THE MISSOURI BOTANICAL GARDEN This experiment was repeated with soil from the ехреті- mental field at Racine. The soil was sifted through а 40- mesh sieve and in this earth cabbage was then planted. All the plants were found diseased at the end of fourteen days, while the plants in a control pot of uninfected soil remained healthy. Plants in a pot of infected soil, unsieved, showed the disease slightly earlier, at the end of twelve days, this difference in time being due probably to the fact that in the sieved soil the organism existed only as spores and, there- fore, took longer to infect than where it grew rapidly from mycelium in the old host tissue. TEMPERATURE Literature.—Before discussing the significance of the tem- perature relation in the case of the attack of F. conglutinans on cabbage, the results obtained in other diseases where tem- perature has proven of pathological importance should be con- sidered briefly for comparison. Upon making a careful review of the literature it was found that our knowledge of this field is very limited and fragmentary, although the importance of temperature is gen- erally recognized. Earle (’02), in a paper on the environ- mental factors concerned with disease, discussed the tempera- ture relations in a general way, pointing out that for health, the plant must have temperatures of the proper degree for growth. Duggar (709) also recognized the importance of temperature in relation to the susceptibility of the host to fungous attack, but in a paper of this nature he could make nothing more than a general statement. Reed (710) ша sim- ilar paper paid more attention to this one of the many envi- ronmental factors involved, and showed that the temperature most favorable for the attack of a fungus is dependent en- tirely on the particular organism under consideration. He cited as examples of this relation the bitter rot of apple which is favored by high temperatures and the leaf curl of the peach which thrives best under cool weather conditions. Klebahn (719, p. 88) stated that, although the temperature is undoubt- 1916] GILMAN—CABBAGE YELLOWS 49 edly an important faetor in the case of diseases caused by the fungi, there has been insufficient work on the question to warrant more than a general discussion of the subject. In spite of this lack of correlated facts on the relation of temperature to plant disease, there are many isolated notes seattered through plant pathological literature, and an at- tempt has been made to bring them together at this time. In order to put them in the best shape for comparison, one with another, it was thought well to arrange them according to the natural grouping of the parasites upon which the ob- servations were made. Schizomycetes.—'The diseases caused by bacteria may be placed first. Halsted (’98) pointed out that the summer of 1894, which was excessively hot in New Jersey, was charac- terized by an outbreak of fire blight due to Bacillus amylovo- rus. Whetzel (’06), from evidence in New York, confirmed this relation but considered that moisture was the more im- portant factor limiting the outbreak of an effective epidemic of this disease. Schuster (712), working with the bacterial rots of potato, showed that at high temperatures (35°C.) saprophytic spe- cies, as, for example, Bacillus fluorescens, might become para- sitie on potato tubers, causing soft rot. Smith (214) noted that in rapidly growing shoots of sus- ceptible hosts the incubation period of Bacillus Solanacearum is shortened in very hot wet weather from 8-10 days to 2-3 days. He attributes the decrease to a difference in suscepti- bility in the host rather than to a change in the virulence of the invading organism. Phycomqycetes.—Perhaps more work has been done on the temperature relations of the Phycomycetes than of any other group of fungi, but the major part of these studies has been with its relation to spore germination. Atkinson (795) found that high temperature was a contrib- uting factor to damping off by Pythium deBaryanum, which view is corroborated by Johnson (714) working at Wisconsin, although neither of these authors show any experimental evi- dence to support their opinion. 4 [Vor. 3 50 ANNALS OF THE MISSOURI BOTANICAL GARDEN Melhus ('11) showed that chilling the conidia of Cystopus candidus to a temperature of 8-10°C. produced the optimum germination and also infection in the case of the radish, and thus proved that this fungus is dependent on such chilling for its best development in its attack on the host. Similar relations have been found to hold true in the cases of other Phycomycetes. From field observations covering a period of twenty years Lutman (711) concluded that Phytoph- thora infestans required a fall in temperature for its best de- velopment on the potato. Melhus (212) showed that the opti- mum temperature for spore germination in this species, both conidia and zoospores, was 8-14°C., thus corroborating the previous field observations. After the fungus has entered, however, he noted in a later paper (18) that the disease was produced more readily at comparatively high temperatures, thus showing that, in this ease at least, there was a difference between the temperatures favorable to parasitie growth, de- pending upon whether the infecting material be spores or mycelium. Reed (712), working with Phytophthor a infestans on tomato plants, found that here again its attack was dependent upon low temperatures. The attack only occurred above the alti- tude of 2000 feet, and then only at times when there were cool nights. Opposed to Phytophthora, Plasmopara Viticola has been found to be dependent on a rather high degree of temperature. Sajó (’01) observed this in 1900 as compared with 1899, the temperature in 1900 being higher throughout the summer than in the previous year. Again, in 1912, Ravaz and Verge (712, 124 2120) showed that for quickest germination a temperature of 22-27? C. was necessary for this fungus and that, since the fungus found water also necessary for infection, the host eould only be attaeked at periods of sustained high tempera- tures and humidity. Istvánffi and Pálinkás (713) showed that not only were Ravaz and Verge correct, but that the de- velopment of conidiophores and conidia from the infected host was also somewhat dependent on these same temperatures. 1916] GILMAN—CABBAGE YELLOWS 51 Ascomycetes.—In the Ascomycetes but little has been done on the temperature relations of the parasitie forms. In Ohio, Selby (799, '04) observed that the leaf curl of the peach, caused by Exoascus deformans, was favored in its occurrence by relatively low temperatures in April, May, and June, the weather in April having the greatest influence. These conclu- sions were based on observations made over a period of ten years, 1893-1903. Duggar (709) showed that the same was true in New York. Pierce ('00) found that similar conditions brought about the attack in California, and attributed the vir- ulenee of the attack to the harmful aetion the adverse weather had on the host, eausing it to be weakened. He also noted that hot dry weather would check an attack which had al- ready started. That Sclerotinia Panacis, the cause of black rot of the gin- seng root, was favored by eold weather was shown by Van Hook (’04). This author found that this disease developed only in the winter, also a time when the roots were in a dor- mant state. Germination of the spores of Sphaerotheca Нитий has been shown by Salmon (’00) to be increased if the spores were previously exposed to cold, especially freezing, temperatures. The germination, however, took place only when the higher temperatures were restored. Sajó (701) observed that Oidium Tuckeri on grapes was favored by subnormal temperature and moisture. Probably one of the first observations of scientific value on the relation of temperature to a particular plant disease was that made on the black rot of grapes by Buchanan in 1850. As is reported by Viala ('87), Buchanan noticed that this dis- ease was worse after a period of hot weather. Viala also made observations on this relation and found that in hot weather (maximum 35-37^C. minimum 18-20°С.) there was a bad epidemie of the trouble. When the temperature fell the disease became checked. His observations covered a period of two years. Later Edson (203), making observa- tions in North Carolina, came to similar conclusions. [Vor. 3 52 ANNALS OF THE MISSOURI BOTANICAL GARDEN That reaction of different parasites to the temperature rela- tions may differ even within a single genus, is well brought out in the genus Glomerella. Неге, on the one hand, is found Glomerella rufomaculans which is dependent on a maximum temperature of 32°C. for the outbreak of an epidemic (Scott, 06), while on the other, Colletotrichum Lindemuthianum (Glomerella Lindemuthianum Shear) is reported by Edgerton (715) as being unable to grow in culture above 31°C. Не shows that this species causes the most severe injury at cool temperatures, infection being inhibited by the summer heat. Fungi Imperfecti.—Little has been done as to this relation in the Fungi Imperfecti. Ravn (700) has shown that in the case of Helminthosporium teres, the attack on barley was con- ditioned on cool temperatures at the immediate time of sprout- ing of the kernel in the soil. Similar conditions held for the stripe disease of barley, caused by Helminthosporium grami- neum. By growing the plants under controlled conditions of temperature, this author was able to show that a temperature of 6.5-14^C. favored the disease, while a temperature of 19- 25°C. practically excluded it from the seed-beds. He showed that the susceptible period for infection was immediately at the time of germination of the seedling, and that plants sprouted in warm temperatures, which were then immediately removed to cool conditions, did not become infected. Bakke (12), in Iowa, showed that the optimum for growth of this fungus in culture was 23-25^C., so that it would appear that the effect of the temperature was one of resistance or escape on the part of the host rather than an effect on the fungus. Further evidence bears out this belief, since Helminthosporium teres can cause a leaf spot in the field at the higher tempera- tures. The question of the temperature relations of the parasitic species of Fusarium will be diseussed later and may be dis- missed here with a brief statement that, as a rule, they seem to require high temperatures for their most virulent attack. In this they appear to be opposed to the closely related genus, Verticillium, which also causes wilt diseases (Wollenweber, 552); 1916] GILMAN—CABBAGE YELLOWS 95 In one other member of the Fungi Imperfecti, Sphaeropsis Ellisii, Petri (713) has observed that the attack was depend- ent on cool humid atmospheric conditions, and the fungus was never seen in warm well-ventilated exposures. 1t is probable, however, that in this disease the limiting factor is moisture rather than temperature. Basidiomycetes.—The temperature relations of the smuts and the rusts have been worked out more exactly than the other Basidiomycetes. Brefeld (795), in experiments with oat smut (Ustilago Avenae), showed that when germinated spores were placed in soil and oats grown therein, 27-30 per cent of the plants became infected at 15^C., while at 7°C. 40-46 per cent were attacked. Tubeuf ('01), working with the same form, found the opposite results when ungerminated spores were used instead of germinated. He showed also that the spores of Ustilago Avenae cannot germinate under 5°C., their minimum for germination being between 5 and 9°С. As is pointed out by Hecke (709), the п im the findings is probably due to the fact that Brefeld germinated the spores before exposing the cultures to the different temperatures while Tubeuf did not. On account of this difference the time of susceptibility of the host was lengthened by the low tem- perature in Brefeld’s experiments, and hence the increased in- fection; while in the experiments of Tubeuf the plants at low temperatures were held below the temperature of germination of the smut spores, and, therefore, the greater infection oc- curred at the higher temperatures. In regard to the stinking smut of wheat (Ustilago Tritici), on the other hand, the minimum temperature for germination of both the wheat kernel (3-4°C.) and the spores of the fun- gus (5°C.) was practically the same, while the maximum for the smut germination (25°C.) was considerably lower than that of the wheat (30—32*C.), so that in this ease the opposite facts were true, as Hecke (709) showed. "Therefore, the in- fection was favored by low temperatures and prevented by high (25°C.), because when the plant grew slowly the length of the susceptible period was increased. Munerati (712) re- ports similar observations on wheat in Italy; early fall and [Vor. 3 54 ANNALS OF THE MISSOURI BOTANICAL GARDEN late spring planting favored the host, while late fall and early spring planting increased infection. Among the rusts a similar relation between spore germina- tion and infection occurs. Howell (’90), working on the clover rust (Uromyces Trifolii), showed that infection would take place only at comparatively low temperatures, the reason given being that it was only at the low temperatures that spore germination occurred; the maximum temperature for both uredo- and aecidiospores was in the neighborhood of 25°С. Marshall Ward (’01) in his notable experiments with the brome rust (Puccinia dispersa) showed that the optimum tem- perature for germination for this form was also at 18°C. Eriksson (’95), in making experiments with rusts, and es- pecially with the germination of spores of different forms, found that chilling the spores in the case of Aecidium Ber- beridis, Peridermium Strobi, Uredo glumarum, and 17. coro- nata accelerated germination when they were brought back to higher temperatures, Johnson (719), working with uredospores of Puccinia graminis, P. rubigo-vera, and P. coronata, showed that their optimum temperatures for germination were 12-17°C.; hence epidemics of grain rusts usually spread at periods of subnormal temperatures. From these observations it is easily seen that the rusts have developed the parasitic habit to a very special degree, adapt- ing the temperatures when there is likely to be dew as those at whieh spore germination will take place, and thus aiding themselves in their attack on the host. Balls (208) has shown in some very careful work on the temperature relation of the Rhizoctonia causing ‘‘sore shin” of cotton that this fungus attacks the cotton plant at 20°C., but not at 83°C. Не checked his work with observations on pure eultures of the organism, and found that at high tem- peratures the fungus secreted, or excreted, an inhibiting sub- stance into the culture fluid which was injurious to the fungus. Whether this same toxie substance prevented that attack on the host is questionable. As to the wood-destroying fungi, Falek (’07) found that Merulius silvester, M. domesticus, M. sclerotiorum, Polyporus 1916] i GILMAN—CABBAGE YELLOWS 55 -vaporarius spumarius, and Verpa bohemica all have а mini- mum temperature for growth of about 3°C.; M erulius sil- vester, M. sclerotiorum, and Polyporus vaporarius spumarius have an optimum of about 25°C., while the optimum for Меги- lius domesticus and Verpa bohemica is at 22° С. Their шах- ima are all at about 30°C. It was noted that these tempera- tures correspond very closely to those of Phycomyces nitens and Mucor Mucedo, as determined for comparison, although each species has a rate of growth that is constant for a given temperature (other factors being equal) and characteristic of that species. With the exception of the genus Fusarium, Ше preceding covers the important work that has been done on the tempera- ture relation of the parasitic fungi, as far as could be ascer- tained. It will be readily seen that the relation of tempera- ture to the attack of a parasite is a complex one and depends entirely upon the individual diseases under observation. To take up now the relations of temperature to diseases caused by Fusarium, Jones (’08) stated in his observations on the damping off of coniferous seedlings, caused by a mem- ber of that genus, that the trouble was facilitated by high temperatures. He was confirmed in this by Gifford (211), working оп Ше same trouble. Wollenweber (713), however, was the first to show this relation in the case of the wilt dis- eases caused by Fusarium. He pointed out that these dis- eases occur most severely in the warmer climates, especially in the tropics and subtropies, but noted the cabbage yellows as an exception to this general rule. Previously, Wolf (710) had noted, in the case of the wilt disease of pansy (Fusarium Violae), that the trouble was found only in July, and then only when the beds in which the plants were growing had been heavily covered with fresh horse manure, both of which facts suggest a dependence of the fungus on high temperature. This author made no mention of temperature, nor were any experiments on this relation reported. Orton (713, 714) in discussing the potato plant and its rela- tion to disease has shown that in this instance Wollenweber’s hypothesis held true, the Fusarium wilt having a southern [Vor. 3 56 ANNALS OF THE MISSOURI BOTANICAL GARDEN range as compared with Verticillium albo-atrum which caused a trouble of almost identieal nature in the northern climates. Neither of these authors made controlled experiments to de- termine, if possible, exact ranges of temperatures. Humphrey (214), working in Washington with tomato blight caused by Fusarium orthoceras App. and Wollenw, and F. oxysporum Schlecht., found that the blight was favored by high temperatures. His statements were based on obser- vations made on experimental plots in 1911 and 1914 at Pull- man, Washington, coupled with the determination of the op- timum temperature for growth of the organism in the labora- tory at 86°F. or 30°C. This author suggested that the light intensity and wind are also factors in bringing about the typical symptoms of the disease. A preliminary report (Gilman, '14) of the relation of tem- perature to the occurrence of cabbage yellows was made at the Philadelphia meetings of the American Phytopathological Society in 1915. The full report of this work is as follows: Field observations.—On the experimental plot at Racine during the summer of 1912 it was observed that the attack of F. conglutinans occurred in the early part of July, when the plants had been set about two weeks. It was noted further that the plants which escaped or withstood the disease at this time remained healthy throughout the rest of the summer. Plants set out after this period were all practically immune. Upon looking up the temperature records of the summer it was found that the attack of the disease followed very closely a period of exceptionally hot weather. Table хуп gives a sum- mary of the observations made at three different times dur- ing the growing period. The strain numbers are those used by Jones and Gilman (715) in the development of a variety of cabbage resistant to yellows. Strains II, ІП, and VI were commercial varieties of Danish Ball-head imported from Den- mark; strains VII, VIII, IX, and X were from seed grown from resistant heads; strain XI was of the Flat Dutch variety imported from Germany; strain XII was commercial Houser; and strain XV, commercial Danish Ball-head. Further de- tails may be found in the publication mentioned above. The. м. = ET 1916] GILMAN——CABBAGE YELLOWS rade. Q 9 y / Degrees Cen ^ 4 ~ № o с о о ы ° Percen tage of Disease Saas ° June July 20 22 24 26 28 JO 2 > 00057 4 D 8 10 ІР 14 18 Je 20 22 24 26 28 30 WNE ` ИТ М Eé. 9 : mm в e шм - -- — tj — ВЕ Н "m Dr? BERRRRSRS f - y | - — dosh 4 4 4 4 $9 4 0 LER |-- m 1.24. i“ ак - a LL TT r1 а PT в m = антта аши GEI UL ж r1 юаш”. з I ш L ша а! | u Г In di - N N ва Ки EEN HE Ñ CET ry H = Lu ка! I I -— i I D Lu EN D H LE - HH =" т LI m. ги ER L E Li Г а! ва каман ай тааш кош ш ши окюшивешми ня им wama) LH ERASEFÓDAAHSTESEAHABERURSERSEEMKESNRAREEE Li ti Е Set eee ЕЕЕ РЕТТТІТІТТІГІТІТІТІТІТІІТІТІІІТТІТІ HL 1 LH D LI H паша шш шж B NL E E е S Cl H H шиа вра CC яти š t И 6 H 58 B ве кә шй ин рк IE BR (8 шиити. ІНІН Bi 711 TT r1 1 ri ct І TI Ц Г Г E LI LI LI LJ LI LI LI LI | LI Li LI H а! Ц I r1 Г m | H ц ri 1 | pa LI ү INI Ц LH RI ананан SES LE rr EI LI LI Li EC aan D LI ER 1 Г LH 11 LI LI LI II Li LI T] 18! D bi H ri LH LI Li ГІ [i 14 LI LI 1 || 11 Ll LE Ll LI A Ц LI IBI H I LI I H rT H I TI П LH im 11 I TI 4 1+ LI SEESRRHSRRREARREE TT IN | 1 Н D H r "^s BEBSNRBESNEEEEEEN LLLE LI | | LI 11 кш 11 RSR и! а! Г] INI ва! Im LL LI LI LI LI яи: 11 CD 11 I | TT ri LL TT Г r а: Li | Ц ] Hid H H SES жашташ шап ЕЕЕ Fig. 13. Comparison of temperature with percentage of disease in field in 1912, [VoL. З 58 ANNALS OF THE MISSOURI BOTANICAL GARDEN plants were set in the field June 24. The disease was begin- ning to show on July 8, when only 0.05 per cent of the plants were yellowed, and increased rapidly to 40.5 per cent on July 29. Of 420 plants set on July 15, after the hottest weather had passed, all remained healthy to August 21, but on account of the late planting did not make heads, and, therefore, were not considered later. The curves given in fig. 13 show the re- lation of the occurrence of the disease to temperature. The temperature data from whieh these were plotted are those pub- lished by the Milwaukee office of the United States Weather Bureau, a distance 25 miles north from the experimental field. TABLE VII SUMMARY OF FIELD OBSERVATIONS ON EXPERIMENTAL PLOT AT RACINE, WISCONSIN, 1912 July 16 July 22 August 20 Total Ке МИМИ! Strain no. o plants | Number | Per cent | Number | Percent | Nu mber | Per cent yellow | yellow | yellow | yellow | yellow yellow СТР ЕРА 45 18 4.0 32 n. 37 82.2 |]: EDENDI 45 3 6.6 24 GR 32 71.1 TU SE 44 2 54.5 35 79.5 43 95.4 VII (a-y)...... 1039 284 27.3 551 3.0 567 54.6 VIII (a & b)... 89 1 Ij 21 3.5 11 12.3 IX (8-116)..... 352 31 8.8 13 6.9 191 54.3 X (101-143).... 625 97 15.5 195 1.2 234 37.4 i S 43 12 27.9 27 62.8 32 74.4 BE 39 5 12.8 18 46 17 43. ( 4 EE 29 8 27.6 19 65.: 20 68.9 ТОШ» 2350 185 20.5 1052 40.5 1184 50.3 Again, in 1913, observations were made in the field on the same plot. The results of these observations are given in table уіп and fig. 14. Besides the strains used in 1912 sev- eral new commercial sorts were introduced. Strains XIII and XIV were Danish Ball-head bred locally, XIII being short- stemmed and XIV long-stemmed; strain XVI was Danish Ball-head grown by the Ferry Seed Company; XVII was All Season; XVIII, Succession; XIX, Volga; XX, Harly Jersey Wakefield; XXI, Copenhagen Market; XXII, Early Summer; XXIII, Charleston Wakefield. While the experience of the 1916] 59 —CABBAGE YELLOWS AN GILM Degrees Centigrade Disease Percentage of Avgust Fig. 14. Comparison of temperature with percentage of disease in ELS 25 27 M9. 3 field in 1913. [Vor. 3 60 ANNALS OF THE MISSOURI BOTANICAL GARDEN previous year was repeated, the relation between temperature and the attack was not as marked as it had been ір 1912. The higher temperatures were sustained for a longer time, and, therefore, the percentage of disease continued to rise through- out the summer. "The plants were set June 24, and no disease was found on July 4. Nevertheless, the main attack occurred in practically the same relation to the hottest weather as it had the previous year. In 1914 the plants were grown on two experimental plots, and in addition to the strains mentioned above, a resistant strain from the Maryland Agrieultural Experiment Station XXIV) was added. The plants were set on June 26, and the disease was first observed on July 16, when 1.5 per cent of the plants showed the typical yellowing. Tables rx and x summarize the observations that were made during this sum- mer. TABLE VIII SUMMARY OF FIELD OBSERVATIONS ON EXPERIMENTAL PLOT AT RACINE, WISCONSIN, 1913 July 19 July 28 September 6 Total Strain no. o plants | Number | Per cent | Number | Per cent | Number | Per cent yellow | yellow | yellow | yellow | yellow | yellow |) METER EUN 50 28 56.0 42 84.0 47 94.0 ІШ АЖЕК ЛИР 46 28 60.8 34 73.9 34 73.¢ EEN 25 54.3 29 63.0 45 97.8 VII (а-у)......| 1243 639 51.4 874 70.3 795 63.€ VIII (a & b)... 96 3 3.1 8 8.3 9 ).4 IX (8-116). .... 248 77 31.0 120 48.4 98 39.5 EE 19 40.4 21 44.7 33 70.2 Eis 17 45.9 24 64.8 24 64. XIII (1-19). 820 301 36.7 489 59.6 748 91.0 XIV (1-11)..... 565 140 24.7 256 45.3 504 ) Muysis 985 556 56.4 707 71.7 827 ) 9,4, 4 Pas 1 25.6 25 58.1 42 m. зад MR 40 17 42.5 26 65.0 32 ( XM un w 28 19 67.8 20 71.4 26 ; P qi EE 42 5 35.7 17 40.5 12 у. ЖОҚ; UP 38 5 39.4 17 44.7 28 ).6 ари 39 20 51.3 23 58.9 37 4.8 EH toss 39 4 35.9 11 28.2 16 41.0 XXIII 33 7 51.5 17 51.5 23 69.7 Total,..... 4485 1961 44.3 2760 51.9 3380 63.5 1916] GILMAN—CABBAGE YELLOWS 61 TABLE IX SUMMARY OF FIELD OBSERVATIONS ON merely орысы ұшы PLOT, HANSCHE FARM, RACINE, WISCONSIN, July 16 July 30 August 17 Total Strain no. o plants | Number | Per cent | Number | Per cent | Number | Per cent yellow | yellow | yellow | yellow | yellow | yellow И S.L... 162 3 1.85 46 | 28.40 54 | 33.33 УШ (ET... 2: 161 2 1.24 40 | 24.83 57 | 35.40 VIIIa (7-35) 810 0 0.0 8 0.99 10 1.23 VIIIb (3-14) 385 0 0.0 8 2.08 19 4.94 264115. 81 0 0.0 Kat Rb 16 | 19.75 JACI 81 0 0.0 8 .87 15 | 18.50 X 135 (2-33)... 782 3 0.38 145 | 18.54 168 | 21.48 X 143 (2-38)...| 1213 2 0.16 215 | 17.72 318 | 26.22 (o Fe ICT 482 27 5.6 374 | 77.59 433 | 89.83 XVI.. 299 5 1.67 144 | 48.16 236 | 78.93 EL a ы 81 11 13.6 67 | 82.7 64 | 79.0 XIII-11 81 1 1:2 69 | 85.2 80 | 98.76 B $E. ue 81 0 0.0 62 | 76.5 74 | 91.35 Beye pi EA 81 1 1.2 14 | 17.3 19 | 23.45 .* 61 Sr 81 0 0.0 6 7.4 5 6.17 Se 4861 55 1.13 1215 | 24.78 1568 | 32.05 TABLE X SUMMARY OF FIELD OBSERVATIONS ON EXPERIMENTAL PLOT, BROESCH FARM, RACINE, WISCONSIN, 1914 July 16 July 30 August 17 ош 5їгаїп of dp Number | Per cent | Number | Per cent | Number | Per cent yellow | yellow | yellow | yellow | yellow | yellow VIIf (5-7)...... 476 ^i 1.47 158 | 33.19 262 5. Уш (2-6)...... 475 4 0.8 172 | 36.2 267 | 56.21 VIIIa (7-35)....| 2352 0 0.0 5 .42 176 7.48 VIIIb (3-18)....| 1175 0 0.0 6 E 136 | 11.57 X 135 (8-21) 1204 0 0.0 315 | 26.16 594 | 49.34 X 143 (2-38) 2379 16 0.67 945 | 39.72 1477 | 62.08 HI 11 238 0 0.00 175 | 73.5 23 97.1 ld PETERE 236 7 2.87 170 | 72.0 225 | 95.8 EE ree 476 137 | 28.7 448 | 94.1 454 | 95.35 XVI. 479 1 0.2 289 | 60.3 426 | 88.9 Total... 9490 172 1.81 2797 | 29.47 4248 | 44.65 Em (VoL. 3 62 ANNALS OF THE MISSOURI BOTANICAL GARDEN A soil thermograph was installed in the experimental plot; the bulb was placed six inches below the surface of the soil, and records were kept covering the growing period. "These show that the temperature of the blaek elay loam, such as is found in Racine and Kenosha counties in Wisconsin, 18 сош- paratively high, the minimum temperature of the soil rarely falling below the minimum for the air, and the maximum tem- perature of the soil, because of the lag, often exceeding that of the air, especially on cold or cloudy days (fig. 15). Тһе same relation between the main attack of the disease and temperature is apparent, although, because the high tempera- tures were maintained throughout July and August as they had been the previous summer, the percentage of disease also increased over a longer period than in 1912. The total per- centage of the disease in this year was less than in the pre- vious years, because plants from the resistant strains were counted with the control, the totals from the entire plot being used. Experimental results.—The experiments to show the rela- tion between temperature and the attack of the fungus were started in 1913. For this phase of the investigation the plants were grown in uniformly diseased soil in two different greenhouses, one of which was kept as near 25°C. as possible and the other at 15-20°C. Im the first experiment three flats of infected soil and one of greenhouse soil (uninfected) were placed in the warm house, and one flat of infected soil and one of uninfected greenhouse soil were placed in the cool house. The flats were planted on October 4, 1913, with two hundred seeds in each flat. The steam was turned on Novem- ber 18, at which time the plants in all the flats appeared nor- mal in their development. On November 25, however, yellows appeared in the flats of infected soil in the warm house. The plants in uninfected soil in both houses remained healthy. Figures 16-18 give an idea of the range of temperatures in the two houses for the entire period during which these experi- ments were made. On December 6 the above experiment was repeated; four flats of infected soil were planted, and two placed in each 1916] 63 GILMAN—CABBAGE YELLOWS L LLLI LLLI ptt + + June July August 20 ge 2 d 0 6 ES H 3 Е E : 5 a mi SS а < ET 2 m а & - = Ba Sech = 2 = = ЕРА Ы - ai ви = в папи в a Ai IER) I H H в CU Pd ене! EI Li H Н--- E š ЦЕНТ ЕН D TYI Ti IB i а! ЕГІТІШІТІІТІС су Ри! LI LI LI LI | | ЕТЕНЕ а III LI II II LI TT BENSE ba LI LI II LE rp LI LI LI 11 if LI LI LI Y an H H rH а! H H Ka UE а! LI H H H H Па S LLEI TI 11 Е == ШВ @ É rr r 4 —— ---- ---- — — —— а BEER DEE Cece HH nananana S HEHE i СЕЕ wa EIS i an OI r cI пип so rT] was Coo H === S "ERIBEIEEE ЫНЫН | SH TE X soto d НЕНІҢ Ч d BEES SERRE БҮ КЕШЕНЕ : КЕНЕНІҢ + LITT] CT LIII 11 I LI ам! 111 кааз 5, п r-r m= I H HHHHTH вана! > " LLL] == РТ] ПІН eg RH S = aan ек 1 : INI 5559559 I NENNEN =: --- | — — — --mmm Mmm - Е = ----- So E pa [ pu mm иинин H = СЕЕ ваши + Н вишкавакее!) SR ШЕ Fig. 15. Comparison of air and soil temperatures with percentage of disease in field in 1914; 5 P D | в 1 ; line; soil temperature broken line. air temperature, solid MI о [ Vor. ANNALS OF THE MISSOURI BOTANICAL GARDEN am рпов “ЕТІ esnog "ott uəso:q OII musk. Met “ет PIBN- DES = re “ІІ Əsno][ ur Əsoq1 пум a ма: 144 — iis uosureduro "ur “Sty LI rrr До shag Ki иә) 622 bijus ә ” 2421 £T HH 7 £1 Ze £o 12761 21 çi £i E et SUD ? Z Ik 62 12 <2 ES HERR Kubnaqay y Gage 494Ш9220 ‘sur рцов “етт Əsnoj[ ‘әп пәңола PII әзпон “етбт “ес 1equiadeq—F 1940120 “етт əsnoH ut E YJIM OTT әзпо ut d Jo шы қыл ыр "9r “SL о ~ 9 “ бие? saashag © м appa eg /г e; Jequia2e(g ладшглем 4290420 1916] GILMAN—CABBAGE YELLOWS 65 house. The yellows appeared in the warmer house on De- cember 29. Each of the seedlings showing the symptoms of the disease was sterilized by placing the entire seedling in hy- drogen peroxide and washing in sterile water. It was then placed on potato hard agar. The fungus grew readily from the stem of the infected seedlings, as is shown in pl. 2, figs. 8-1. The flats of infected soil were interchanged, and after replanting on January 29, the results were found to be the same; that is, the plants in the warmer house showed the disease, while those in the cooler house remained healthy. The first disease symptoms were observed on February 10. During the above experiments the temperatures were as constant as they could be made in a greenhouse where the steam supply was regulated by means of an automatic thermo- regulator. Of course, the heat on very sunny days was much greater than desired, but this factor could not be controlled, as shading caused too rapid elongation of the plants and a consequent susceptibility to damping off. It was found, how- ever, that the soil temperatures were fairly constant, being from 23 to 26°C. in the warmer house and from 12 to 16°C. in the cooler house. These determinations were made directly by placing the thermometer bulb two inches below the surface of the soil and after the mercury had come to rest making the reading. In one experiment the number of plants used in the trial was noted, and the percentage diseased after an exposure of three weeks calculated from actual count. The experiment was started January 29, 1914. The trial consisted of seven pots of infected soil, and two pots of normal greenhouse soil for controls. Five pots of infected soil were placed in the warmer greenhouse and two in the cooler house. One pot of the normal greenhouse soil was placed in each house. The disease was found first on February 10, and the plants were pulled and counts made on February 21. If the experiment had been continued, doubtless all the plants in the warmer house would have been destroyed, as they had been in the other experiments. Table xr gives the results. 5 3 [ VoL. ANNALS OF THE MISSOURI BOTANICAL GARDEN 66 “әш uƏsoiq II ósnog (oui рпов 'eureij j -pjoo {PTET “с eunf-er YLW II Əsno] ut esoqj чум әше1;-р|09 out ur seinjeieduioj [ros jo uosrreduio;) S P E IE 62 42 <2 (2 мү "әш pijos “II esuopg “әш uoyouq 911 әзпо {Ф161 “г Хен -ет YIU “ІІ евпон “ur Əsoq) дим II әвпоң ur вәлпүеләйшә) Jo повъавфшод 7% ot uf a4bogg иә) sa 7 app. әрриб шә seeibeg 1916] GILMAN—CABBAGE YELLOWS 67 TABLE XI AMOUNT OF YELLOWS PRESENT AFTER AN EXPOSURE OF THREE WEEKS TO HIGH AND LOW TEMPERATURES After three weeks’ exposure Pot Total Soil Tem- o о No. P perature plants diseased diseased еба. Ube C. 73 27 37 pil 7. i ror 5 К 25; C. 69 25 38 S Iu ntected. a coo m ет РС, 60 16 21 ek EE Mes 2w C. 54 20 37 ес WE E DuC. 44 18 41 Let EE E 15-20? C. 60 0 0 7 succ) pr ИТТЕР MI 15-20? C. 49 0 0 8 |Emmected ..... AV ERËN 6 2b. C. 119 0 0 Есей e. 15-20? C. 58 0 0 Again, on February 21, the effect of transplanting normal plants to infected soil at different temperatures was tried. Fifty normal plants were placed in two flats of infected soil February March 22 „24. 83 27 2 2 3 4 f$ 6 7 40 // /2 E 74 /$ > Ki ~ 9 to ° I ° Percentage of disease Degrees Centigrade ч 9 Fig. Ка Comparison of soil temperatures in House Ile and those in House p^ with the occurrence of yellows in the respective houses; House Ile, bro- n line; House Ila, solid line. E one flat placed in each house. Controls consisted of ten normal plants from the same flat as the above, placed in nor- mal greenhouse soil in two pots, one pot in each house. Soil [Vor. 3 68 ANNALS OF THE MISSOURI BOTANICAL GARDEN temperatures were taken daily. The results are not conclu- sive, as the temperature in the cooler house rose to 21°C. on the sixth day and remained high for two days, nor did it go back to below the temperature at which infection took place. Nevertheless, the symptoms appeared in the warmer house March 4, three days before the plants in the eool house showed any sign of the trouble, and the cooler soil retarded the advance of the fungus proportionately, as may be seen from the curves (fig. 20). In further experiments on this temperature relation a cold- frame was used as a means of maintaining cooler conditions, for the greenhouses were all too warm, due to the increased intensity of sunlight, especially at midday. The north side of the potting-house, where the sun was excluded, made it pos- sible to carry these cultures still further into the spring, and in all cases the results were the same. In the experiment in which the cold-frame was used, a soil thermograph of the type manufactured by Julien P. Friez was installed. The bulb was imbedded four inches in the soil, and the temperature of the soil and air were recorded through- out the experiment. The plants were started on March 25 in six flats of uniformly infected soil, three of which were placed in the greenhouse at 25° C. and three in the eold-frame. Two pots of greenhouse soil, planted to cabbage, were used as controls in each case. The disease appeared first in the green- house on April 4, ten days after planting. Seedlings were plated from the diseased flats and from the flats in the cold- frame on April 9, and in all eases the diseased seedlings showed the fungus growing from the stem, while the controls remained sterile. On April 13, photographs were made of two of the flats—one from the cold-frame showing the healthy condition of the seedlings, and one from the greenhouse show- ing the ravages due to the attack of the fungus (pl. 2, fig. 7). The temperature records show that there was an in- crease in temperature with the advance of the season, and it was due to this increase in temperature that the attack oc- curred. The curves (fig. 19) do not show this fact well, as they are a record of the maximum and minimum only and do 1916] GILMAN—CABBAGE YELLOWS 69 not show the duration of temperature in any one day. The records themselves, while they cannot be presented here, show this increase more markedly—for the length of time of the higher temperatures increased—as the spring gave way to summer conditions. In any case the cooler condition pre- vented the attack of the fungus for at least a month. In the experiment in which the flats were placed on the north side of the potting-shed, three flats of diseased soil were used. Two were placed on the north side of the potting- shed, and the third in the warm house. "The flats were planted April 9, and yellows appeared in the flat in the greenhouse on April 17, while none was found, up to May 26, in the flats kept outside. Soil temperatures covering this period are shown by the curves in fig. 21. /0 /2 14 /6 /8 20 28 24 26 28 J 2 4 8 /o ІР 14 /6 |8 20 EE) TH HFE BESESRERRbRSER EC rt r1 EC) IN | ri | ПП LI LI D EC H TI Cl ТТЕП H IN в! EC LH ri d 3o mmmmm HH —- E | —— | БЕКЕБЕЕЕГЕКЕТІ 5 PH Н H HHH < ГЕ BEREEE г Er, 2 AR Н H \ ааа! + an a CONC " © 20 SEH в = == E GER e к? иии TI r Н H " 5 222224 H H Е ma Cl im) g! и! =" Q Se EECH AHH H S % Sierck) LH Li O O LI a % 10 CL ri Ц ишн шша жож еш) < ва! | H A E 4 rH LLLELELALLTI Ov аа! N Г] MI VI Hu Co Y rH ПИТА Ini M H Coo ma Hi LX ІШ! N H COCO eer ШІ Li H LI H I H этн шт ДАЛ ТІТТЕГІТІІТТІІ су cot H LI H а! H LLLELEEELLELLLELELELLELLTU Hi а! г ai H H LLLLELHILLLCELLLEEELHLELLETTET II LH Г LI T$ rH и! SURES PORES Re | Fig. 21. Comparison of soil temperatures in House Пе with those on north side of potting-shed, April 10-May 20, 1914; House Пе, broken line; north side of potting-shed, solid line. Further work was undertaken at the Missouri Botanical Garden to try to determine the lowest point at which the at- tack may occur. To this end two glass incubators were set up, and the temperature was controlled by an electric thermo- regulator, so that it varied but a degree or two at the most. One incubator was set up in the laboratory at a west window, and the other was placed on the north side of the building. The temperature of the incubator in the laboratory was kept [Vor. 3 70 ANNALS OF THE MISSOURI BOTANICAL GARDEN at 22-24^C., this being the lowest constant temperature that could be procured in the room. The incubator outside was set at 16°C., but on account of the wide variation in atmos- pherie temperatures it was not possible to keep it constant at this point. At times the temperatures reached as high as 21°C., and at other times as low as 10°C. In spite of this variation no yellows occurred in the incubator which was out- side until some days after all the plants in the warmer incu- bator had been attacked. Twenty pots of soil from the experi- mental field at Racine were placed in the warm incubator and twenty similar pots were placed in the cool incubator. The pots were put in the incubators on November 5, 1914, and in four days the plants in one pot showed wilting. The plants were sixteen days old when they were submitted to the trial. Previous to this time they had been growing in the green- house which was kept at 12-14°C. On November 11 yellowing was apparent in all the pots except the controls, some of the plants also showing wilting. In the outside incubator but two plants showed a slight yellowing and there was no wilt- ing. Damping off due to Rhizoctonia was rather extensive in some of the pots because of the relatively high humidity eonditions. Counts were taken on November 14 of the plants in the warm ineubator, and seedlings from both ineubators were plated on potato hard agar. The results were as fol- lows: At 22-24" C. F. conglutinans was isolated from plants from every pot of the infeeted soil, while none could be isolated from the plants in the normal greenhouse soil. When the plants from eight pots of infected soil, growing at 10-16°C., were plated F. conglutinans was isolated from but one; the fungus was not isolated from the eontrols. In the twenty pots at 22-247 C. there was a total of 104 plants, seven of which re- mained healthy, while in the incubator at 10-162 C. there was a total of eighty-eight plants, only one of which showed the disease even after they had been left until December 1 in the incubator. Table хп summarizes the results obtained on this temperature relation. 1916] GILMAN—CABBAGE YELLOWS TX TABLE XII SUMMARY OF RESULTS OF EXPERIMENTS TO SHOW THE RELATION TEMPERATURE TO ATTACK OF THE FUNGUS Extent of trial Approximate Incu- average temperature | Condition bation Infected soil Uninfected soil іп °С. of control „period at incep- in days | Higher | Lower | Higher | Lower tion of temper- | temper- | temper- | temper- disease ature ature ature ature Higher Lower 13 3 flats.| 1 flat 1 flat 1 flat 22° 16-20° | Healthy.. 21 1 flat 1 flat 1 pot 1 pot 22° 16-20? | Healthy*. 30 10 pots.| 12 pots.| 1 pot 1 pot 14-16? | Healthy.. 4 20 pots.| 20 pots.| 3 pots.| 3 pots.| 22-24? 16° | Healthy”. 23 3 flat fl lebar. [2 flat... 4-18? | Healthy.. 12 3 flats.| 1 flat wat...) 2 Gat... 25° 4-18° | Healthy.. 12 7 pots.| 5 pots.| 2 pots.| 2 pots. 25° 4-18° | Healthy.. 8 2 flats flat..| 2 pots.| 2 pots. 25^ 4-16? | Healthy.. 10 З flats.| З flats.| 2 pots.| 2 pots. 30° |4-16° | Healthy*. *In these cases the controls in infected soil at the lower temperatures became diseased later, due to a rise of temperature above the point at which they were able to resist the disease. In a further experiment seeds were planted on October 20 in pots of infected soil and allowed to stay in the green- house. The soil temperature was 10-16°С., and while a little disease appeared from time to time after December 1, 1914, the attack was very light and very few of the plants suf- fered. When on February 2, five pots of these plants were taken to a greenhouse whose air temperature was 28-30°C., yellows appeared in virulent form in three days and all but three plants were dead on February 20. Plate 1, fig. 3, shows typical pots from this experiment. It was repeated February 28 with similar results. The fact that high temperatures caused the yellowing of the cabbage when the plant was attacked by F. conglutinans hav- ing been clearly established, the next point was to find, if pos- sible, whether the fungus entered the host at the lower temper- ature or not. The first experiments were made by plating from the plants in the lower temperatures, especially from parts of the roots of plants grown in infected soil. Although the plates were made in the same manner and with the same [VoL. 3 12 ANNALS OF THE MISSOURI BOTANICAL GARDEN care as in the ease of the yellowed seedlings grown at higher temperatures, at no time was F. conglutinans isolated from the roots of these plants. Controls from roots grown at the higher temperatures showed the fungus, as has been pointed out in previous experiments. Experiments were therefore instituted to test whether any such relation might be indi- eated by indirect methods. The first experiment was started on April 21, 1914, at which time twenty pots of infected soil were planted to cab- bage, and all were placed on the north side of the potting- shed where a low temperature could be maintained. "They were kept here until June 6, sixteen days, when all but two were placed in House Пе which was being kept at approxi- mately 25°C. On June 9, after the pots had remained in the warm house for three days, pairs of pots were removed to the cooler temperature at intervals of two days until June 15, after which date a pair was removed each day until June 19. On June 30 yellows appeared in the two pots removed on the last day, June 19, but none was found in any of the other pots. This experiment showed that in this case the yellows appeared in the same length of time as it usually took to ap- pear in a warm house, and would lead to the opinion that there had been no infection at the low temperature, or if the plants had been attacked, that they were able to recover under favorable conditions for growth. Coincident with this last experiment, moreover, twenty pots of infected soil planted to eabbage were placed in House lle, and each day two pots were removed to the cooler tem- perature. No yellows appeared in any pots removed in the first eight days, but in all those removed subsequently yellow- ing was found on June 2. Controls in the warm house showed the first symptoms on June 1, one day earlier than those on the outside. The cooler temperature, therefore, checked the disease in most cases, but where it had gone too far, the only effect was a slight lengthening of the period of incubation. Later observations on this point do not seem to confirm these results. It will be noted that in the experiments car- ried on at the Missouri Botanieal Garden, when the plants 1916] GILMAN—CABBAGE YELLOWS 73 were first grown in the greenhouse and then placed in the in- cubator at 22-24^C., the disease appeared in but four days, a period that was shorter than had been noted in any other experiment. This trial was repeated on March 9, and again the seedlings showed the disease in four days, on March 13. Hight pots of seedlings were used, and the disease ap- peared in all the pots on the same day, although not all the seedlings in any one pot were yellow at this time. Previous to the appearance of the yellows, platings made from the roots by the hydrogen-peroxide method gave negative results in all eases. Further, the roots were washed out of the soil and examined carefully under the microscope, but no hyphae of the fungus were observed until after wilting or yellowing had begun. The rotting usually began at the tips of roots near the surface of the soil, and progressed toward the main roots and stem. The only explanation that seems applicable to these conflicting results is that, because the temperature in the greenhouse at the Garden is slightly higher than that found on the outside of the potting-shed, the fungus may enter to a limited extent, but cannot affect the host unfavorably ex- cept at the higher temperature, while at Madison it was un- able to gain any sort of a foothold. This view is further supported by the fact that a few plants grown in diseased soil in this greenhouse, after a long period of time showed yel- lows, as previously mentioned. Because the small number of hyphae found in any single diseased stem seemed insufficient for the blocking of the pas- sage of water to the leaves of the diseased plant, some pre- liminary work was undertaken to find, if possible, whether mechanical or chemical killing of the stem might bring about symptoms in the leaves similar to those produced by the fun- gus, and especially with regard to the production of a toxic substance to which the symptoms might be ascribed. To test this question six plants of cabbage were cut on one side with a scalpel so that half of the stem was removed for a distance of 0.5 em. The plants were about two weeks old and growing rapidly. The cut surfaces were covered with paraffin to prevent too rapid drying of the tender tissues. Two 7 ЫБ. D я, = ` [Vor. 3 74 ANNALS OF THE MISSOURI BOTANICAL GARDEN plants showed wilting in nine days but the others all remained upright and turgid, having completely recovered. There was no discoloration of the leaves in connection with the wilting. In a later experiment with older plants, the entire stem of each plant was killed for a distance of 3 em. from the surface of the ground by allowing it to stand in alcohol for three minutes. After nine days, wilting appeared in one of the five plants, the lowest leaves drooping first, but with no dis- coloration such as occurs under the influence of fungous at- tack, nor falling of the wilted leaves. A second plant suc- cumbed on the twelfth day, but again there was rapid wilting with no loss of green coloring matter. By the eighteenth day all the plants had wilted, but even where the injury had been least and the wilting slowest there was no discoloration or falling of the leaves. The experiment was repeated with older plants in March, 1915, with similar results. The wilting al- ways took place without discoloration of the leaves, nor did any of them drop before the entire head was wilted. Further work was started, therefore, to see whether the fun- gus could produce in pure culture any substances toxic to eab- bage. For this study two Erlenmeyer flasks of half-liter ca- pacity, each containing 100 сс. of Uschinsky’s fluid, were in- oculated with a virulent culture of F. conglutinans. After two weeks the fungus-felt was filtered from the solution by means of a pressure filter, and the solution, after dilution to 500 ce., was poured in two glass tumblers, in which germinated eab- bage seedlings were then placed. Controls of Uschinsky's fluid diluted 2:5, tap water, and Pfeffer's full nutrient were used in connection with the experiment. Difficulty was ex- perieneed in getting the plants to start because of the desic- cation of the young cotyledons. By placing the plants in an incubator under humid conditions, the plants growing on tap water and Pfeffer’s solution grew fairly well, but on Uschin- sky’s fluid, neither on that in which the fungus had been grow- ing nor on the sterile fluid, was it possible to get any growth, indicating that some other media or methods will have to be used. Further work on this point is being pursued. 1916] GILMAN—CABBAGE YELLOWS 1e Discussion.—Exaetly why the raising of the temperature should bring on this disease is still not clear, but in view of our present knowledge some correlation should be made be- tween the relations found and the other work that may shed light on this point. First, it should be pointed out that many of the so-called vascular parasites behave in a very similar manner toward temperature. As Smith (714) has shown with Bacillus Solanacearum, Humphrey (714) with Fusarium or- thoceras, and the present investigation with F. conglutinans, high temperatures facilitate the destruction of the host. To what extent this destruetiveness may be attributed to changes in the parasite and to what extent to changes in the host plant, it is diffieult to determine. Smith and Humphrey both are inclined to consider the changes in the host the primary fac- tors concerned, and as will be pointed out, the same opinion may be taken in the case of the cabbage disease. Nevertheless, the change in the fungus must be looked into also. Among diseases of plants that are partially dependent on temperature relations for their oceurrence, in many of the cases the relation is not one of loss of virulence on the part of the fungus but a limitation in the temperature range of ger- mination of the fungous spores. This sort of limitation was best illustrated by the work of Melhus (212, 713) on Phytoph- thora infestans as related to the potato blight. This author showed that, although the spores germinated only at low tem- peratures, the mycelium which wintered over in the tuber, at- tacked the new shoots from such tubers only at high tempera- tures. Other cases of similar nature, where the temperature for spore germination differed from that of mycelial growth, are found among many of the obligate plant parasites. Exam- ples that might be cited are Cystopus candidus, Plasmopara Viticola, Ustilago Avenae, U. Tritici, Uromyces Trifolii, Peridermium Strobi, Puccinia graminis, P. rubigo-vera, P. dispersa, and P. coronata. That Fusarium conglutinans is not dependent on germina- tion temperatures for its destructive attack is clearly shown by the fact that germination occurs readily at 17°C., which temperature is close to the lower limits of its destructiveness [Vor. 3 76 ANNALS OF THE MISSOURI BOTANICAL GARDEN in the case of cabbage. Moreover, it grows readily at tem- peratures much lower than this. The raising of the tempera- ture merely increases the rapidity of the growth of the fungus and, therefore, as far as this investigation is concerned, the high temperature from the standpoint of the fungus aids its destructiveness by increased spread in the soil, and more rapid development in the vascular system after it has once entered. Other possible relations, such as that of production of toxic substances, remain to be worked out. From the host standpoint the effect of temperature is much more complicated. Appel (715) in his discussion of leaf roll in potato considered excessive transpiration of prime im- portance in bringing about this condition whether the cause of the trouble was parasitic or not. The symptoms of the dis- ease in the cabbage indicate that the phenomena involved are very similar to those concerned with the annual autumn fall of leaves from woody plants. The discoloration (yellowing in the diseased plants), the formation of an abscission layer, and finally the fall of the leaves are in all ways comparable. Hence the same physiological changes within the plant are probably taking place. From this point of view then, the work of Molisch (286) and Varga (711) on the relations of environmental factors to the fall of leaves gives a basis for an explanation of the symptoms from the host standpoint. Molisch showed that a slow but continued decrease of water content of the fundamental tissue of the leaf led to the for- mation of an abscission layer and finally fall of the leaf. He further found that this loss of water might be brought about by increased transpiration or by decreased absorption or con- duction from the roots to the leaf. Temperature influenced leaf-fall, both indirectly through its effect on transpiration, and directly by bringing about the formation of the abscission layer. Leaves fell at 17-22°С. more rapidly than at 1-10°C. when other conditions were equal. Varga studied the relation of temperature to leaf-fall more exactly and found that, as a rule, low temperatures lowered transpiration and thereby set up a stimulus to leaf-fall, but that if the abscission layer had been formed through other influences, higher tempera- ET 1916] GILMAN—CABBAGE YELLOWS 77 tures within limits, eaused а more rapid fall of Ше leaves. These facts give a possible explanation of the results found with cabbage yellows. The hyphae of F. conglutinans in the fibro-vascular bundles cause a constant but slow drain on the water content of the plant, which causes the formation or the beginning, at least, of the formation, of the abscission layer. High temperatures, in addition to causing increased growth of the fungus, raise the transpiration and also stim- ulate leaf-fall; thus all the factors are cumulative in their effect. The reason that mechanical and chemical injuries to the stem did not cause similar symptoms may be explained by the fact that the plants wilted before sufficient time was given for the formation of the abscission layer and, therefore, the difference in symptoms. This theory also concurs with that of Humphrey in regard to the tomato blight, but a large amount of work is still necessary before it will be completely proven. SuMMARY Cabbage yellows is a wilt disease of cabbage caused by Fusarium conglutinans Wollenw. The fungus is a facultative parasite living in the soil, from which, under certain conditions, it becomes destructive to cabbage. The fungus has a high optimum temperature and is very resistant to drying—both in pure culture and in the soil. Inoculation experiments with Fusarium conglutinans in pure culture caused the disease in a large percentage of the trials. Control plants remained entirely free from the yellows. Fusarium conglutinans was recovered from inoculated dis- eased seedlings and again produced the disease upon inocula- tion. Variation in virulence of the cultures and in susceptibility of the host caused many artificial inoculations to be unsuc- cessful, resulting in less than 100 per cent infection. [VoL. 3 78 ANNALS OF THE MISSOURI BOTANICAL GARDEN Mechanical or chemical injury to the stem of the host caused wilting, but neither yellowing nor dropping of the leaves such as is found in diseased seedlings. The characteristic symptoms are dependent on a tempera- ture of about 17-29°С, or above for their occurrence. Lower temperatures (12-16°C.) under controlled conditions pre- vented the occurrence of the trouble in the greenhouse. Observations made in the field during the summers of 1912, 1913, and 1914 bore out this relation between the occurrence of the disease and high temperature. In conclusion, the writer wishes to express to Dr. L. R. Jones, at whose suggestion this investigation was undertaken, and to Dr. B. M. Duggar, under whom it was completed, his sincere appreciation of the many valuable suggestions and helpful criticisms given during the progress of this work. He is further indebted to the support of the Wisconsin Ex- periment Station for the opportunity of conducting the initial stages of the work and to the Missouri Botanical Garden for the completion of the work upon the problem. Graduate Laboratory, Missouri Botanical Garden. LITERATURE CITED Appel, O. Seri Leaf roll diseases of the potato. Phytopath. 5:139-148. 1915. — Wollenweber, H. W. (210). Grundlagen einer Monographie der Gattung decem (Lino. K. biol. Anst. f. Land- u. Forstw., Arb. 8:1-207. NU Atkinson, с ch (79 ix Damping off. Cornell Univ. Agr. Exp. Sta., Bul. 94:233- —6. f. 55. um 2 19 p^ 2). The m A. e 2 авн teres Saee.). Ia. Acad. Sci., Proc. Balls, W. L. (708). Ex ën iig us Bot. 22:557-591. f. 1-4. 1908. — de A id Untersuchungen aus dem Gesamtgebiete der Mykologie 11:27- De re y Hedwigia 19: p. 191. 1880. Duggar, B. M. ('09). The тр ur conditions of growth ыт. Lg eme o fungous diseases. Mass. Hort. Soc., Trans. 1909:51-66. no Е. 8. дни А. рн ^ disease in plants. N. Y. Bot. GE m 3:195- 202. a z Y ыс och Effect of temperature on Glomerella. (Science N. 8. Fason ri e gr^ ). The black rot of grapes in North Carolina. N. C. Agr Exp. Sta., Bul. 185: 133- 156. 1903. Eriksson, J. (9 5). Ueber die Forderung = SE durch Кайе. Centralbl. f. Вак. II. 1:557-565. 1f. 1916] GILMAN—CABBAGE YELLOWS 79 Falck, R. ('07). Wachstumsgesetze, Wachstumsfaktoren und Temperaturwerte der holzzerstórenden Mycelien. Móller's Haussehwamm-forschungen. Heft 1:53-152. 1907 Gifford, C. M. (11). The damping off of coniferous seedlings. Vt. Agr. Exp. Sta. Bul. 157:143-171. pl. 1-4. f. 1-10. 1911. ¿sg J. C. (714). The relation of temperature to the infection of cabbage by arium conglutinans Wollenw. (Phytopath. 4:404. 1914). i =a B. D. ('98). Influence of wet weather upon parasitie fungi. (Am. Assoc. Adv. Sei, Proc. 47:416. 1898). Harter, L. L. (709). Fusarium wilt of cabbage. Science N. S. 30:934. 1909. ------, (712). Diseases of cabbage and өтеле 2 ш and their control. U. 8. Dept. Agr., Farmers’ Bul. 488:18-21. f. 4. 1912. pos x я 9). Der тии von Sorte und Mese auf De Steinbrand- Zeitschr. f. Landw. Versuchsw. ()sterr. 19:46-66. 1909 SENS J. K. ('90). pe 277 rust. Cornell Univ. Agr. Exp. Sta., Bul. 24 129-139. f. 1-8. Humphrey, H. B. (’14). Studies on the relation of certain species of Fusarium to the tomato be + the Pacific Northwest. Wash. Agr. Exp. Sta., Bul. 115:1-22. pl. 1 Istvánffi, Zeg and Pálinkás, G. (’13). Etudes sur le чи s ei е” Ann. l'Inst. Cent. Ampelol. Roy. Hongrois. 4:1-125. pl. 9. f. 1 Johnson, E. C. (712). Cardinal temperatures for the 21 of uredospores of cereal rusts. (Phytopath. 2:47-48. 1912). pou ch AS 14). The са of damping-off disease in plant beds. Wis. Agr. ә Res. Bul. 31:29-61. f. 1-12. 1914. Jones, L. (708). The E off of coniferous seedlings. Vt. Agr. Exp. Sta., Rept’ 20:342-347. 1908. ——r (713). Тһе possibilities of disease resistance in cabbage. (Phytopath. 3:71. 1913). —— (14). Progress in developing disease-resistant cabbage. (Ibid. 4:47— 48. 1914 4). —— (7143). Third progress report on Fusarium-resistant eabbage. (Ibid. 4:404. 1914 Ж — and Gilman, A i E 15). Тһе control of cabbage GES сер disease resistance. в. Agr. Exp. Sta., Res. Bul. 38:1-70. f. 1- — EE 20718), Beck der ЕКЕН: Phytopathologie. pp. 1-147. 1-24. Berlin, 1912. ежен С. E. (718). Comparative studies of certain disease TEES species of Fusarium. Me. Agr. Exp. Sta., Bul. 219:203—9258. f. 89-1 1913. Lutman, B. Е. (711). Potato owns and the weather. Vt. Agr. Exp. Sta., Bul. 159:248-996. 911. Manns, T. Е. (711). Two recent important XL diseases of Ohio. Ohio Agr. Exp. Sta., Bul. 228:262-275. f. 2-14. xg I. E. (711). Experiments on spore M and infection in certain cies of Oomycetes. Wis. Agr. Exp. Sta., Res. Bul. 15:25-91. pl. 1-10. 1911. -----> (12). The factors governing germination and infection with Phytoph- thora infestans. (Phytopath. 2:46. 1912). — N (713). н myeelia of Phytophthora and other related species. (Ibid. 3:70. 1913). s [Vor. 3 80 ANNALS OF THE MISSOURI BOTANICAL GARDEN Molisch, E Ще 4 Кач. über Laubfall. Sitzungsber. d. k. Akad. d. , Wien 931:148-184. 1886 Munerati, O. (712). Sulla — € del ka oon per la earie in rapporto al tempo di semina. Ассай. Lincei, Rendie. 21':875-878. 1912. Norton, J. B. 8., and imo UPC 6, in. Control of SE H^ and diseases of Mad mida Md. Agr. Exp. Sta., Bul. 115 Orton, W. А. (713). Environmental AM in the ue. of Solanum tuberosum. Washington Acad. Sei, Jour. 3:180-190. f. 1-3. 1913. ——— ——, (714). Potato Ki = roll, and related diseases. U. 5. Dept. Aer, Bul. '64 :1-16. pl. 1-2. Petri, 1. (213). Disseecamento dei rametti di Pseudotsuga Douglasii Carr. prodotto da una varietà di Sphaeropsis Ellisii Sace. Ann. Мус. 11:278-280. 13. cuore m B. (700). eges leaf curl: its nature and treatment. U. S. Dept. , Div. Veg. Phys. and Path., Bul. 20:22-30. 1900 Ravaz, L. and Verge, G. h m pon de frm du mildiou. Prog. Ag "T. et Vit. 55:485-488. 19 ——————, — —- M, (123). Influence de la reno sur la germination des eonidies du mildiou. bid. 55:170-177. 1912 —————, ——————, (7126). Sur les conditions 4” apparition des eonidiophores du mildiou. Ibid. 55:206-300. 1912. Ravn, F. K. (700). Nogle Helminthosporium-Arten og de af en ам ұлы бу gdomme hos Byg og Havre. Bot. Tidsskr, 23:101-321. pl. 1-2. f. 1 1900. Reed, G. M. ('10). The influence of EEN conditions upon the devel- opment of plant diseases. Mo. State Bd. Hort., Bul. 31:1-14. 1910 — H. S. (712). Does ыа сыы infestans cause tomato blight? Phy- opath. 2:250-252. 1912. Ru K. (201). seh big ir Ansprüche von ety Tuckeri und Peronos- pora viticola. Zeitschr. Pflanzenkr. 11:92-95. 19 ———, (20 Verschiedene ред gees der schädlichen Pilze. Prometheus 13:132-135, 154—157. 190 Salmon, E. S. (700). The strawberry mildew. Roy. Hort. Soc., Jour. 25:132- 138. f. 35—36. 1900. agg " (712). Zur Kenntnis der Bakterienfaule = 7” K. biol. . Land- u. Forstw., Arb. 8:452-492. pl. 5. f. 1 m ч M. (706). The — of "e bitter-rot. U. 8. 5 Së Bur. PL , Bul. 93:1-33. pl. 1-8. f. 1 Bey, A N D. (799). Variations in the amount of leaf curl of tne peach (Exoas- sd Әле in the light of weather conditions. Soc. Prom. Agr. Sei, pue. 20:98-104. 1899. , (704.) Peach diseases III. Ohio Agr. Exp. Sta., Bul. 148:57-58. 1904. Smith, E. F. (’99). The fungous infestation of agricultural soils in the United States. Scientif. American, Suppl. 48:19981. 1899. , ат А wilt disease of cotton, watermelon and cowpea. U. 8. Dept. Agr. Div. Veg. Phys. and Path., Bul. 17:41-42. 1899. -----, (1 Baeteria іп нох to plant diseases. Carnegie Inst. Wash- ington Publ. 973:174-206. 1914. ги L. (13). The fungi which cause plant disease. р. 654. New York, 1916] GILMAN-——CABBAGE YELLOWS 81 Tubeuf, = F. von (’01). Studien tiber die Brandkrankheiten des Getreides und ihre Beka ümpfung. K. biol. Abt. f. Land-u. Forstw., Arb. 2:179-349. pl. 8, 7: 1-19. Van Hook, J. M. (20 Diseases of ginseng. Cornell Univ. Agr. Exp. Sta., Bul. 219:181-182, f- 5а 1904. Varga, O. (71 Beitrage zur Kenntnis дет Beziehungen des Lichtes und der od a zum Laubfall. Oesterr. Bot. Zeitschr. 61:74-88. Vaughan, R. E. (714). A method for the differential — of i ads and host cells. Ann. Mo. Bot. Gard. 1:241-242. 1914 Viala, P. (87). Les maladies de la vigne. pp. 245-270. Paris, 1887. Ward, M. (201). The bromes and their brome rust. Brit. Assoe. Adv. Sei, Rept. 71:836-839. 1901. теш, H. H. (?06). The blight canker of apple-trees. Cornell Univ. Agr. 1906. ( . Sta., Bul, 236:103-138. f. 49-84 Wolf, Е. A. (710). A Fusarium disease of the pansy. Mycologia 2:19-22. 1910. Wollenweber, H. W. (713). Ji» nrc Welkekrankheiten der Kulturpflanzen. Ber. d. de 1913 ut. bot. Ges. 31:17-34. f. 1. 1913 oods, A. Е. (799). The destruction of chlorophyll by oxidizing enzymes. 1899. (799) Centralbl. f. Вак. II. 5:745-754. $i Studies on the Fusarium problem. Phytopath. 3:24—50. pl. 5. [VoL. 3, 1916] 82 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 1 Fig. 1. Results of inoculation a = greenhouse with pure cultures. Pot No. 1. Infected soil, steri Pot No. 2. Sterilized soil in EE with pure culture of F. conglutinans. Pot No. 3. Soil from c m untreated. Madison, Wisconsin, July 1913. Fig. 2. Results of ino pended on in Y^ garden with pure cultures. Pot No. 1. Infected soil, sterilized. Pot No. 2. Sterilized soil inoculated with pure culture of F. conglutinans. Pot No. 3. Soil from infected т untreated. Madison, Wisconsin, July 1 1913 Fig. 3. Effect of temperat ure on p E of F. conglutinans on cabbage. Pot No. 1. Uninfeeted soil in cool hou Missouri Botanical — March 1, 1915. Fig. 4. Diseased cabbage plant show 5. typical one-sided bending of leaf and loss of lower leaves. Madison, Wisconsi ваза Е PLATE 1 ANN. Mo. Bor. GARD., Vor. 3, 1916 Fig. 3 GILMAN—CABBAGE YELLOWS COCKAYNE, BOSTON [V or. 3, 1916 84 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 2 Figs. 5 and Comparison of rate of germination of resistant and com- mercial varieties of cabbage under the same conditions. Fig. 5, commercial sort ; fig. Be p Baca gio Botanieal Garden, November 5, 1914. Fig. 7. t of temperature on the attack of F. 'conglu utinans on cabbage. On left, flat from “cold Kate on right, flat from House IIe. Madison, Wiscon- sin, April 19, 1914 Figs. 8 ,9, 10, and 11. Effeet of temperature оп Mes attack of F. conglutinans on cabbage. Fig. 8, plants from temperature control in cold-frame; fig. 9, plants from soil control in House Ile; fig. 10, plants we Fiat o. lin House Ile; fig. 11, plants from Flat No. П in House Ile. s Nos. I and II and the temperature control = bas orn infeeted E "Soil control was uninfected greenhous i. M a ie ae 9 Branched "abba age t, one о DE, showing yellows, while Fig. 12. other, BC, remains healthy. Platings Ken from marked points amd results shown in fig. 13. Madison, Wisconsin, May 19, Fig. 13. Plate made from plant shown, in De 12. Note that pieces B and C from healthy branch remained sterile. Mad , Wisconsin, May 21, 1914 ig. 14. Stems of infected cabbage adi on potato 5. Ae ` Note Ed growth from vascular bundles and ends of cut stem. Madi , Wisconsin. 15 den three EE of stem from each of gom healthy cabbage plants grown in sterilized soil; fig. 16 shows the same from three diseased plants grown in soil that had been sterilized and inoeulated ar pure culture of F. conglutinans. Madison, Wisconsin, July 16, 1913. PLATE 2 VoL. 3, 1916 4 ANN. Мо. Вот, GARD., —— - —— т i e к € š 42 uH: r da % ; , GILMAN—CABBAGE YELLOWS COCKAYNE, BOSTON MONOGRAPH OF THE NORTH AND CENTRAL AMERICAN SPECIES OF THE GENUS SENECIO—PART II J. M. GREENMAN Curator of the Herbariwm of the Missouri Botanical Garden Associate Professor in the Henry Shaw School of Botany of Washington University Бест. 6. Aurer Rydb. 86. Aunzi Rydb. Bull. Torr. Bot. Club 27:173. 1900; Greenm. Monogr. Senecio, I. Teil, 22, 23, 29, 30. 1901, and in Engl. Bot. Jahrb. 32:18, 19, 25, 26. 1902. Herbaceous perennials, glabrous or in the early stages floceose-tomentose and more or less glabrate except in the axils of the leaves and occasionally at the base of the stem; stems erect or ascending, 1 to 10 dm. high, one to several from a common base or rootstock; leaves variable, the lowermost petiolate, rotund-ovate, oblong-ovate, obovate to narrowly oblanceolate, entire, crenate or dentate to more or less lyrate; stem-leaves petiolate to sessile, pinnatisect to entire, usually reduced towards the eymose inflorescence; heads radiate or discoid; achenes glabrous or hirtellous along the angles. Sp. 33—80 KEY To THE SPECIES A. Stems 1 to 10 dm. high, ereet or nearly so, simple or branched; heads one to many, diseoid (except in S. pauciflorus var. fallax); achenes glabrous. a. Plants small, 1.5 dm. or less high; lower leaves subentire or i dentat a. Lower leaves subentire.............. 33. S. fedifolius B. Lower leaves dion CAR EET 34. S. Fernalds b. Plants larger, 1.5 to 10 dm. high; leaves thin in texture, the до erenate- dentate Issued July 7, 1916 NoTE.—The present paper is continued from Ann. Mo. Bot. Gard. 2:573-626. 1915. ANN. Mo. Вот. GARD., Vor. 3, 1916 (85) [Vor. 3 86 ANNALS OF THE MISSOURI BOTANICAL GARDEN a. Stem-leaves .5 to 3 em. broad, usually with “. сие, divisions and narrow sinuses. I. Heads pm 1. Lower leaves obovate......... 48a. S. obovatus var. elongatus 2. Lower leaves ovate.......... 35. 8. pauciflorus IL. Heads radiate ..4.25- rr 85a. 8. pauciflorus var. fallax В. Stem-leaves 1 to 6 em. broad with re- mote divisions and deep rounded TTT EE E 6. 8. idahoensis с. Plants 1 to : bor high; leaves thickish in texture, the er egi crenate to un- equally lobate. Kafe ek 37. S. debilis B. Stems erect or ascending, 5 to 4 dm. high, simple or branched; heads dréie usually solitary; achenes glabrous or hirtellous. 8. 2. mostly irregularly lobed ог sub- lyra а. green BIDEN Au SEENEN 38. S. hi ангина enes glabrous .................. 39. S. resedifoli b. Leaves mostly or all undivided. ves thickish in rang ; heads 8 I. Plants glabrous or nearly so..... 40. 8. ovinus 11. erg white-tomentose at the base and in the leaf-axils. 1. Heads 12 to Bm high; in- volueral braets 21 .......... 41. 8, conterminus 2. Heads 8 to 10 mm. high; in- volueral bracts 13 .......... 42. 8. hesperius В. ш thin in texture; heads 8 to 12 m. high, ES Plants of northern United States and Canada. 1. iks leaves coarsely dentate. 43. 8. Newcombei 2. Lower leaves crenate-dentate. 44. 8. subnudus II. Plants of Мехісо............... 46. 8. Rosei Y. Leaves reniform to oblong-obovate, thickish in texture; heads 14 to 20 mm. igh. I. Leaves reniform ............... 46. 8. Porter, II. Leaves subrotund to oblong- ПЪОМОЦОС oce EE 47. 8. Soldanella C. Stems usually erect, e to 7 dm. high E or branched; nch heads eommonly sev any, usually radiate (discoid іп 8. oe А” уат. elongatus, 8 . rubricaulis var. aphanactis and rarely in 8. pauperculus). 1916] GREENMAN—-MONOGRAPH OF SENECIO 87 a. Leaves thin in texture, not succulent in the living state. a. Basal leaves obovate, subrotund to oblong-elliptic, usually glabrous... Basal leaves rotund-ovate, oblong- ovate to oblong- води cordate to abruptly narrowed at the base, gla- brous or glabrate. I. Lower leaves rotund-ovate, usually Xo 2. Heads e to many. * Plants of Mexico. de segment o stem-leaves broader than long; achenes s hispidulous ........ tt Terminal segment of stem-leaves not broader than теді achenes glabrous . icio idus ES western United Sta ..“е............. II. Lower Sasa ovate to oblong- lanceolate, KA cordate or abruptly narrowed at the base. 1. Margins of lower leaves cre- nate- а to od T * Lower leaves, some of them, perd cordate. em T oblong- lanceolate ......... tł Basal pus ovate. || Margins lower leaves Eg serrate with ineurved teeth. || Margins of lower leaves more coarsely and тоге saliently d. o. Bracteoles broad, obtuse... оо. Bracteoles n acute . ** Lower leaves abruptly cuneate at the base, not cordate. t Eastern species. | Upper leaves pin- nately divided ..... 48. 49. 50. 61. 55. Зда. 8. obovatus S. Cardamine 8. cyclophyllus 8. quebradensis S. Раттеш S. aureus S. Robbinsit S. pseudaureus S. pauciflorus var. fallax S. Burkei 8. gaspensis [Vor. 3 88 ANNALS OF THE MISSOURI BOTANICAL GARDEN ШТ EE а in- cised-s ae tt Western | species. II diea of stem- leaves broad ...... 2. pisi пу of lower leaves entire r near * Rays usually orange-red or saffron-eolored ...... ** Rays lemon-yellow. tems closely ces- РОВ о неа tt yea x closely ces- pi | Upper leaves con- spicuously dilated ted into a broa amplexieaul base ... Y. Basal leaves NE. -ovate to Beggen often shallowly cordate and leaves more or less persistently wie. tomentulose, rarely glabro I. Pla the mountain of Arizona and Ne Mexico; achenes glabrous ............... к - . Plants of the low country and prairie throughout central United 1 achenes usually hispid- д. Basal Ge oblanceolate (except in S. пада var. firmifolius), gradually n at the base, glabrous or Sub € m! subglaueous ............ II. Plants not at all glaucous. LA —— rather densely and per- ently tomentose аб the Tun dS usually numerous. bo . Stems but slightly — at the base; heads compar tively few * Head 5 to 9 mm. high; stems not flexuous. { Eastern species ...... tt igre species ....... ** Heads 10 to 13 mm. high; stems т flexuous ..... Сл ° су i 60. e к су » съ = су 5 68. S. Crawfordii S. quaerens S. platylobus S. crocatus 8. aquariensis S. dimorphophyllus S. Farriae S. Hartianus S. plattensis S. Willingii 8. Smallii 8. pauperculus S. flavovirens S. multnomensis 1916] b. аас ари usually thick or firm іп texture, or less succulent in the living state. a. Lower leaves ovate to obovate, sub- entire to pi dentate. I. Lower leaves broadly oval to elliptic- oe subentire to cre- nate-dentat 1. Stems 2.5 to 5 dm. high.. 2. Stems usually lower, 1 to 2.5 GI c. l Geek cs. Cee T II. Lower leaves pe ee en- tire to sharply den 1. Lower leaves ee dentate. 2. Lower leaves entire or dentate towards the apex only....... В. Lower leaves mostly oblanceolate, entire or dentate chiefly towards the apex I. Plants 1 to 4 dm. high; cymes open, more or less flat-topped. L eio tending to be leafy; leav FE 2 em. wide, achenes ta da SE gien not leafy; leaves nar- .5-1 em. wide; achenes ing eege II. imi less than d hig ymes elose, somewhat rounded. . bo Y. Lower leaves larger, ovate to oblan- eeolate, subentire to coarsely and sali- ently den tate with subeartilaginous oc? Т, а ich or a slightly чер tate, what gla agami plan of the "United Забава . Leaves usually eme dentate, not glaucous; plants of Mexico. = M 74. x E x P 80. GREENMAN——MONOGRAPH OF SENECIO 89 S. laetiflorus S. Suksdorfii S. rubricaulis S. eymbalarioides S. acutidens S. tridenticulatus 8. Тағай 8. anacletus 8. toluccanus [Vor. 3 90 ANNALS OF THE MISSOURI BOTANICAL GARDEN 33. S. fedifolius Rydb. Bull. Torr. Bot. Club 27:183, pl. 5, fig. 7. 1900, and КІ. Colo. 397. 1906; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. S. discoideus Nelson in Coulter & Nelson, Manual Cent. Rocky Mountains 583. 1909, in part. An herbaceous perennial; stem slender, 1.5 dm. high; basal leaves petiolate, blade ovate or broadly oval, 1 to 2 em. long, entire or wavy margined; stem-leaves small, pinnately di- vided into short linear-oblong segments; heads 2 to 3, about 6 mm. high, discoid; involucral bracts commonly 13, lanceolate with membranous margins; achenes glabrous. Distribution: mountains of Colorado. Colorado: South Park, coll. of 1871, Canby (N. Y. College of Pharmacy Herb.), TYPE. 34. 8. Fernaldii Greenm.' A small herbaceous perennial, glabrous or slightly tomen- tose in the axils of the leaves, more or less purplish; stem erect, 1 dm. or less high, rising from an oblique rootstock; lower leaves petiolate, ovate to obovate-cuneate, including the petiole 1.5 to 3 em. long, 1 to 1.5 em. broad, sharply dentate; stem-leaves pinnatisect, the uppermost reduced to mere bracts; head solitary, discoid, 10 to 12 mm. high; involucre campanu- late, calyeulate; bracts of the involucre about 21, linear-lanceo- late, 7 to 8mm. long, a little shorter than the numerous flowers of the disk, purple; achenes glabrous. Distribution: western Newfoundland. Specimen examined Newfoundland: dry limestone barrens, upper slopes, and tablelands, Table Mountain, alt. 200-300 m., 16 Aug., 1910, 18enecio Fernaldii Greenm. sp. nov., herbaceus perennis glabrus vel in axillis foliorum parum tomentosus plus minusve purpurascens; eaule erecto 1 dm. minus alto; foliis inferioribus petiolatis ovatis vel obovato-euneatis petiolo incluso 1.5-3 em. longis 1-1,5 em. latis acute dentatis, superioribus Speso gradatim reductis; capitulo solitario 10-12 mm. alto diseoideo; involucri squamis circiter 21 lineari-lanceolatis 7-8 mm. longis; flosculis disci numerosis; aus glabris. —On dry lim wore barrens, upper "er and tablelands of Table Mountain, Newfoundland, 200-300 m., 16 A Se Fernald $ Wiegand 4188 (Gray Herb., Sg in Mo. Bot. Gard. Herb), TYP 1916] GREENMAN—MONOGRAPH OF SENECIO 91 Fernald Ф Wiegand 4188 (Gray Herb., photograph in Mo. Bot. Gard. Herb.), TYPE. 35. S. pauciflorus Pursh, Fl. Am. Sept. 2:529. 1814, and ed. 2, 1816; Schlecht. in Linnaea 10:90. 1836; DC. Prodr. 6:431. 1837; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Piper, Contr. U. S. Nat. Herb. 11:597. 1906; Piper & Beattie, Fl. Northwest Coast 387. 1915. S. aureus ?. discoideus Hook. Fl. Bor. Am. 1:333. 1834; Torr. & Gray, Fl. N. Am. 2:442. 1843, in part. S. aureus var. borealis Gray, Syn. Fl. N. Am. 1:391. 1884, and ed. 2, 1886, in part, i. e., as to S. pauciflorus Pursh in syn- onymy. S. Lemberti Greene, Pittonia 3:89. 1896. S. indecorus Greene, Fl. Franciscana 470. 1897. S. aureus pauciflorus Britt. in Britton & Brown, Ill. КІ. 3 :480. 1898. S. discoideus Britt. in Britton & Brown, Ill. Fl. 3:479, fig. 4042. 1898, and ed. 2, 3:544, fig. 4626. 1913. S. elongatus Howell, Fl. Northwest Am. 1:379. 1900, in part, not Pursh. S. aureus Britt. in Britton & Brown, Ill. Fl. 3:544. 1913, in part, i. e., as to S. pauciflorus Pursh in synonymy. An herbaceous perennial, glabrous or with a white-floccose tomentum in the axils of the leaves and in the inflorescence; stems one to several from a common base, erect or ascending, 1 to 10 dm. high; lower leaves petiolate, ovate-rotund to ovate- oblong, 1 to 8 em. long, 1 to 4.5 cm. broad, crenate-dentate, glabrous on both surfaces, their petioles equalling or twice ex- ceeding the blade; stem-leaves sublyrate to pinnatisect, the uppermost sessile and much reduced; inflorescence a few to many-headed corymbose cyme; heads discoid, 10 to 12 mm. high; involuere eampanulate, calyculate; bracts of the in- voluere usually about 21, linear-laneeolate, acute or acutish, more or less purplish; flowers numerous; achenes glabrous. Distribution: usually in bogs and wet places, Labrador, eastern Quebec, northern Michigan, across the continent to the [Vor. 3 92 ANNALS OF THE MISSOURI BOTANICAL GARDEN Rocky Mountains, northwest to Alaska, and south to northern California. Specimens examined: Labrador: specimen from the Pursh herbarium, ex Herb. Dickson (Phil. Acad. Nat. Sci. Herb.) ; without locality, '' Le- conte” (Phil. Acad. Nat. Sci. Herb.) ; about Hokhak and Heb- ron, fratres Morav. (Gray Herb.); northern Labrador, Lat. 58°, coll. of 1873, Anspach (Mo. Bot. Gard. Herb.) ; wet crevices of rocks, Nachvak, 1 Aug., 1884, Bell 14778 (Geol. Surv. Can- ada Herb.) ; Long Point, Aug., 1892, Waghorne 4 (Geol. Surv. Canada Herb. and Mo. Bot. Gard. Herb.) ; Capstan Island, Aug., 1893, Waghorne 9 (U. Б. Nat. Herb.) ; near Fortean, coll. of 1894, Waghorne 20 (Mo. Bot. Gard. Herb.); Rama, Lat. 58°50’, July, 1894, Sornborger (Gray Herb.) ; Rama, 20-24 Aug., 1897, Sornborger 67 (Gray Herb. and U. S. Nat. Herb.) ; Rama, 15 July-20 Aug., 1897, Stecker 67" (Mo. Bot. Gard. Herb.) ; without locality, Dr. Morrison (Kew Herb.). Quebee: Lake Petitsikapau, Hamilton River, 26 June, 1894, Low 5104 (Geol. Surv. Canada Herb.); sandy shores of River Ste. Anne des Monts, Gaspé, 19 Aug., 1882, Macoun 14808 (Geol. Surv. Canada Herb.) ; ealeareous alpine meadow, Table-top Mountain, Gaspé Co., alt. 1000-1125 m., 7 Aug., 1906, Fernald € Collins 261 (U. S. Nat. Herb.) ; on limestone con- glomerate cliffs, peak west of Baptiste Michaud’s, Bie, Rimouski Co., 16 July, 1904, Collins & Fernald (Gray Herb.) ; wet meadow, Bie, 22 July, 1910, Williamson 1418 (C. S. Wil- liamson Herb.) ; arbor-vitae swamps, Carleton, Bonaventure Co., 24-27 July, 1904, Collins, Fernald & Pease (Gray Herb.). Ontario: wet gravelly places, Nipigon River, 2 July, 1884, Macoun 14782 (Geol. Surv. Canada Herb.) ; bogs north of Port Arthur, 6 Aug., 1912, Williamson 2148 (Phil. Acad. Nat. Sci. Herb.); Fort William, 24 July, 1912, Williamson 1722 (Phil. Acad. Nat. Sei. Herb.) ; near Lake Superior, Macoun 53 (Mo. Bot. Gard. Herb.). Michigan: Keweenaw Peninsula, coll. of 1863, Robbins 121 (Gray Herb.) ; Keweenaw Peninsula, July, 1890, and 7 July, 1916] GREENMAN—MONOGRAPH OF SENECIO 93 1915, Farwell 776 (Gray Herb. and Mo. Bot. Gard. Herb.) ; Champion, July, 1889, Hill (Gray Herb.). Rocky Mountains: “Grand saline, R. M. E. side," Burke (Gray Herb.). Montana: Gallatin Valley, near Bozeman, alt. 1615 m., 7 July, 1896, Flodman 908 (Mo. Bot. Gard. Herb., Greene Herb., апа U. S. Nat. Herb.) ; wet shady places, Gallatin River, 14 July, 1905, Blankenship 292 (Mo. Bot. Gard. Herb., Phil. Acad. Nat. Sci. Herb., Field Mus. Herb., and U. 8. Nat. Herb.). Wyoming: Little Goose Cañon, Sheridan Co., 1 July, 1901, Nelson 2383 (Mo. Bot. Gard. Herb.) ; Middle Ten Sleep Creek, Big Horn Co., 1 Aug., 1901, Goodding 465 (Gray Herb., U. 8. Nat. Herb., and Mo. Bot. Gard. Herb.). Idaho: Forks of St. Mary’s River, alt 1100 m., 3 July, 1895, Leiberg 1158 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.). Alberta: below Pipestone Summit, Rocky Mountain Park, 6 July, 1904, Macoun 65018 (Gray Herb. and Geol. Surv. Can- ada Herb.) ; Blind Valley and Lakes, alt. 2130-2440 m., 6 July, 1906, Brown 414 (Phil. Acad. Nat. Sci. Herb.) ; head of Snake River, 5 Aug., 1911, Riley 32 (U. S. Nat. Herb.). Maekenzie: Fort Franklin, Mackenzie River, Richardson (Geol. Surv. Canada Herb., photograph in Field Mus. Herb. and in Mo. Bot. Gard. Herb.), part of түре of S. aureus var. discoideus Hook. Yukon: Ft. Selkirk, 18 July, 1899, Tarleton 134 (С. 5. Nat. Herb.); vieinity of Summit and Middle Lakes, coll. of 1898, Bolton (U. S. Nat. Herb.) ; moist glades, Red Mountain, 17 July, 1899, Gorman 1114 (U. S. Nat. Herb.) ; Moosehide Moun- tain, Dawson, 14 July, 1902, Macoun 78974 (Field Mus. Herb.) ; damp woods, West Dawson, 16 July, 1902, Macoun 78975 (Field Mus. Herb.); Finlayson River, Lat. 61°N., Dawson 14776 (Geol. Surv. Canada Herb.). Alaska: south bank of Forty Mile Creek, Yukon River, 13 July, 1893, Funston 126 (U. S. Nat. Herb.) ; near Knik, Oct., 1913, EE (Mo. Bot. Gard. Herb.) ; Kenai, 18-20 Aug., 1904, Piner 4216 (U.S. Nat. Herb.). [Vor. 3 94. ANNALS OF THE MISSOURI BOTANICAL GARDEN British Columbia: Moose Lake, 14-24 Aug., 1911, Riley 22 (U. S. Nat. Herb.) ; Kicking Horse Lake, alt. 1675 m., 15 Aug., 1890, Macoun 14771 (Geol. Surv. Canada Herb.); Kicking Horse Valley, near Field, alt. 1230 m., 20 June-25 July, 1906, Brown 487?" (Phil. Acad. Nat. Sei. Herb.) ; Ottertail Drive, near Field, July, 1905, Farr 814, 820 (Univ. Penn. Herb. and Field Mus. Herb.) ; Wapta Lake, 4 Aug., 1904, Macoun 65019, 65020 (Gray Herb. and Geol. Surv. Canada Herb.) ; Carbonate, alt. 825 m., 7 July, 1904, Heacock 185 (Gray Herb., U. S. Nat. Herb., Phil. Aead. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.) ; Upper Spillmacheen Valley, alt. 1980 m., 3 Aug., 1904, Petersen 440 (Gray Herb., U. S. Nat. Herb., Mo. Bot. Gard. Herb., and Phil. Aead. Nat. Sei. Herb.) ; Cornwall Hills, 28 July, 1894, McCoy 5100 (Gray Herb. and Geol. Surv. Canada Herb.); Skagit Valley, 28 June, 1905, Macoun 69360 (Gray Herb.) ; Maclennan River, branch of Columbia River, 31 July, 1898, Spreadborough 19727 (Geol. Surv. Canada Herb. and Greene Herb.) ; Chilli- wack Valley, 22 June, 1901, Macoun 26683 (Gray Herb., Geol. Surv. Canada Herb., and Mo. Bot. Gard. Herb.), 26684 and 26685 in part (Gray Herb. and Geol. Surv. Canada Herb.); alpine rivulet, Goldstream, alt. 1675 m., 25 July, 1905, Shaw 1012 (U. S. Nat. Herb.); Alberni, Arrowsmith Trail, Van- eouver Island, 27 June, 1907, Rosendahl 1971 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Wolf Creek, Strathcona Park, Vancouver Island, 10 Aug., 1912, Macoun 83192 (Geol. Surv. Canada Herb. and Mo. Bot. Gard. Herb.) ; Kasaan Mountain, Queen Charlotte Island, 7 July, 1901-02, Newcombe 69 (Field Mus. Herb.). Washington: Deming, Whatcont Co., 30 June, 1898, Flett 852 in part (Piper Herb.) ; Big Meadows, six miles west of Ione, 6 Aug., 1902, Kreager 428 (Gray Herb., U. S. Nat. Herb., and Piper Herb.) ; Mt. Constitution, Dreas Island, Aug., 1892, Henderson 2312 (Gray Herb.). California: Truckee, 16 June, 1901, Williamson (C. S. Wil- liamson Herb.) ; Mt. Dana, coll. of 1866, Bolander 6021 (Field Mus. Herb.) and 6021 in part (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Lassens Peak, July, 1896, Austin 365 (U. S. Nat. 1916] GREENMAN—MONOGRAPH OF SENECIO 95 Herb. and Mo. Bot. Gard. Herb.) ; Pine Creek, Lassen Co., 9 July, 1894, Baker £ Nutting (Greene Herb.), түре of S. indec- orus; Sierra Co., coll. of 1874, Lemmon 127 (Mo. Bot. Gard. Herb.) ; Sierra Nevada Mountains, coll. of 1875, Lemmon (U. 5. Nat. Herb. 48716); Soda Springs, Tuolumne Co., July, 1894, Lembert 171 (Gray Herb.); Yosemite Region, coll. of 1893, Lembert (Gray Herb. and Greene Herb. in part); Soda Springs, Mt. Conness, 6 Aug., 1890, Harford (Greene Herb.), TYPE of S. Lemberti. To this species are also to be referred two specimens without record of locality, namely, one from the collection of Nuttall, presented by Elias Durand, 1866 (Gray Herb.), and one from the Bernhardi collection (Mo. Bot. Gard. Herb.). Var. fallax Greenm. Contr. U. S. Nat. Herb. 11:597. 1906, and in Piper & Beattie, Fl. Northwest Coast 388. 1915. Similar in stature and in foliage to the species; heads ra- diate; ray-flowers 10 to 12, rays yellow; disk-flowers 50 to 60; achenes glabrous. Distribution: occurring occasionally with the species. Specimens examined: Ontario: Silver Islet Beach, 4 Aug., 1914, Williamson 2075 (C. S. Williamson Herb.) ; Port Arthur, 6 Aug., 1914, William- son 2148 (C. S. Williamson Herb.). Michigan: Mainland Park Harbor, Isle Royal, 15-16 Aug., 1912, Williamson 2312 (Phil. Acad. Nat. Sci. Herb.). Alaska: along the Yukon River, near Ft. Yukon, coll. of 1881, Bates (U. S. Nat. Herb. 48813, 48814, and Greene Herb.). Washington: Deming, Whatcom Co., 30 June, 1898, Flett 852 in part (Piper Herb.). California: Mt. Dana, coll. of 1866, Bolander 6021 in part (Gray Herb., U. S. Nat. Herb, and Mo. Bot. Gard. Herb.) ; Yosemite Region, coll. of 1895, Lembert (Gray Herb. and U. S. Nat. Herb.) ; Mt. Goddard, alt. 3050 m., Най € Chandler 660 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; along the North Fork of Kern River, Sierra Nevada, 25 Aug., 1891, Coville ё Funston 1708 (U. S. Nat. Herb.). [Vor. 3 96 ANNALS OF THE MISSOURI BOTANICAL GARDEN 36. S. idahoensis Rydb. Bull. Torr. Bot. Club 27:185, pl. 6, fig. 5. 1900; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. A rather stout herbaceous perennial 4 to 7 dm. high, glabrous or slightly white floccose-tomentulose in the axils of the leaves; stem branched from near the base, rather leafy in the lower portion, nearly naked towards the inflorescence, stramineous to somewhat purplish; leaves 2.5 to 14 em. long, 1 to 6 em. broad, mostly pinnately parted into oblong or some- what cuneate subincised divisions with broad and deep round- ed sinuses; inflorescence terminating the stem and branches in a several to many-headed corymbose cyme ; heads discoid, 10 to 12 mm. high; involucre eampanulate, ealyeulate; bracts of the involucre commonly 21, linear-lanceolate, 7 to 9 mm. long, acutish, more or less purplish-tipped; flowers numerous; achenes glabrous. Distribution: Idaho, northwest into British Columbia. Specimens examined: Idaho: in meadows at Granite Station, Kootenai Co., 30 Aug., 1892, Sandberg, MacDougal & Heller 803 (Gray Herb. and Phil. Acad. Nat. Sci. Herb.), со-түРЕ. British Columbia: Griffin Lake, 6 July, 1889, Macoun, with- out number (Gray Herb.). A species very near the preceding from which, however, it differs in the branehing of the stem and by the large broad stem-leaves with eut divisions and deep rounded sinuses. 37. S. debilis Nutt. in Trans. Am. Phil. бос. 7:408. 1841; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Blankenship in Mont. Agr. Coll. Sci. Studies 1:102. 1904. S. aureus var. discoideus Torr. & Gray, Fl. N. Am. 2:442. 1843, in part, i. e., as to S. debilis Nutt. in synonymy. S. aureus var. borealis Gray, Syn. Fl. N. Am. 1:391. 1884, and ed. 2, 1886, in part. S. flavovirens Rydb. Bull. Torr. Bot. Club 27:181. 1900, in part, as to plant of Greene. | | 1916] GREENMAN—MONOGRAPH OF SENECIO 97 S. nephrophyllus Rydb. Mem. N. Y. Bot. Gard. 1:446. 1900, and in Bull. Torr. Bot. Club 27:183. 1900. S. discoideus Nelson in Coulter & Nelson, Manual Cent. Rocky Mountains 583. 1909, in part, not S. aureus var. dis- coideus Hook. An erect herbaceous perennial, glabrous throughout or slightly floceose-tomentulose in the axils of the leaves and along the midrib on the upper surface of the young leaves; stems 1 to 5 dm. high; leaves thickish in texture; basal leaves petiolate, subreniform to ovate-oblong, 1 to 6 cm. long, 1 to 4 em. broad, entire, crenately and unequally lobate-dentate ; petioles variable in length from 1.5 em. to nearly 2 dm. long; stem-leaves petiolate or sessile, sublyrate to pinnately di- vided into remote linear-oblong or subeuneate obtusish, often unequal, divisions and deep rounded sinuses; inflorescence terminating the stem in a few to several-headed corymbose всуше; heads 7 to 10 mm. high, discoid; flowers numerous; achenes glabrous. Distribution: Montana and Colorado, west to Idaho and Oregon. Specimens examined: Montana: meadows, Big Blaekfoot River, 13 July, 1883, Canby 34 (Gray Herb.), and 203 (Phil. Acad. Nat. Sci. Herb.) ; Melrose, 6 July, 1895, Shear 5011 (U. S. Nat. Herb.). Wyoming: Laramie Plains, alt. 2590 m., 20 July, 1884, Sheldon 73 (U. S. Nat. Herb.) ; Laramie, 28 July, 1889, Greene (U. S. Nat. Herb. and Field Mus. Herb.); Laramie hills, 17 July, 1897, Nelson 3404 (Mont. Agr. Coll. Herb.) ; Laramie Plains, 21 July, 1898, Osterhout (Field Mus. Herb.); wet banks, City Springs, Laramie, 8 Aug., 1901, Nelson 8599 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; Laramie River, along the Medicine Bow Mountains, coll. of 1856, H. Engelmann (Gray Herb. and Mo. Bot. Gard. Herb.); wet meadows, Centennial, 27 July, 1900, Nelson 7722 (Mo. Bot. Gard. Herb.); boggy draws, Centennial, 27 July, 1902, Nel- son 8685 (Gray Herb., U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.); Wind River bottom, 28 July, 7 [Vor. 3 98 ANNALS OF THE MISSOURI BOTANICAL GARDEN 1882, Forwood (Gray Herb. and U. S. Nat. Herb.) ; Meadow Creek, 9 Aug. 1894, Nelson 906 (Gray Herb. and Greene Herb.); Bridger's Pass, coll. of 1856, H. Engelmann (Gray Herb.) ; Soldier Springs, Aug., 1891, Nelson 177 (U. S. Nat. Herb.). Colorado: Rocky Mountains, coll. of 1861, Parry 19 (Phil. Acad. Nat. Sei, Herb.); Rocky Mountains, Lat. 39°41’, coll. of 1862, Hall € Harbour 332 in part (Mo. Bot. Gard. Herb. and Gray Herb.) ; Lake John, 19 Aug., 1898, Shear € Bessey 3990 (U. S. Nat. Herb.) ; near High, 19 Aug., 1898, Shear Ф Bessey 4005 (U. S. Nat. Herb.). Idaho: grassy bog, One Thousand Springs Valley, alt. 2040 m., 7 Aug., 1895, Henderson 3671 (U. S. Nat. Herb.). Oregon: ‘‘ Plains of the Oregon, near the Wahlamet,’’ Nutt- all (Gray Herb.), түре. 38. S. hyperborealis Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Ann. Mo. Bot. Gard. 1:264. 1914. S. resedifolius Hook. Fl. Bor. Am. 1:333, pl. 117. 1834, in part, not Less.; DC. Prodr. 6:347. 1837, in part; Torr. & Gray, Fl. N. Am. 2:445. 1843, in part; Gray, Syn. Fl. N. Am. 1?:390. 1884, and ed. 2, 1886, in part; Macoun, Cat. Canadian РІ. 267. 1884, in part, 1. e., as to plant of Richardson. Stems erect or nearly so, one to several from a perennial base, 1 to 2 dm. high, simple or branched, white-tomentose at the base and in the leaf-axils; lower leaves obovate and erenately margined to sublyrate or pinnately divided with rather remote obtusely dentate divisions, including the petiole 4 to 10 em. long, 1 to 2.5 em. broad; stem-leaves sessile, more or less pinnatisect, the uppermost reduced to lance-attenuate entire bracts; heads solitary or few terminating the stem and branches, 8 to 12 mm. high, radiate; involucre campanulate, sparingly calyculate, glabrous; bracts of the involuere about 13, lanceolate, acute, 5 to 6 mm. long; ray-flowers 10 to 12, rays yellow; disk-flowers rather numerous; achenes hirtellous- puberulent along the angles. Distribution: Arctic northwest America. 1916] GREENMAN—MONOGRAPH OF SENECIO 99 Specimens examined: Mackenzie: ‘‘on limestone at the mouth of the Bear Lake River, and about Fort Norman and Fort Franklin,’’ Richard- son (Gray Herb. ex Hooker, Torrey Herb., and Geol. Surv. Canada Herb. 14872 in part), co-ryPE; west shore of Great Bear Lake, June-Aug., 1900, Bell 22937 (Geol. Surv. Canada Herb.). 39. S.resedifolius Less. in Linnaea 6:243. 1831; Ledeb. КІ. Rossica 2:631. 1844-46, in major part; DC. Prodr. 6:347. 1837, in major part; Torr. & Gray, Fl. N. Am. 2:445. 1843, in part; Gray, Syn. Fl. N. Am. 1?:390. 1884, and ed. 2, 1886, in part ; Macoun, Cat. Canadian РІ, 267. 1884, in part; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. Cineraria lyrata Ledeb. in Mem. Acad. Petersb. 5:576. 1818; Reichb. Ic. Bot. 2:1, pl. 101. 1824; Hook. & Arn. Bot. Beechey's Voy. 126. 1832. A low herb; stems one to several from a perennial base, simple or branched, .5 to 2 dm. high, glabrous or slightly tomentose, particularly in the axils of the leaves; lower leaves petiolate, rotund-ovate, sublyrate or irregularly pinnately di- vided, crenate to sharply dentate; upper stem-leaves sessile, more or less pinnatisect, often reduced to lance-attenuate en- tire bracts; heads solitary or few, radiate, about 1 em. high, including the rays 2 to 2.5 em. in diameter; bracts of the caly- culate involucre linear-lanceolate, 6 to 8 mm. long, acuminate, acute, often purplish; achenes glabrous. Distribution: Alaska and Siberia. Specimens examined: Alaska: St. Lawrence Island, Chamisso (Gray Herb., Ber- lin Herb., and Kew Herb.); Behring Strait, U. S. North Pacific Exploring Expedition, 1853-56, Wright (Gray Herb.) ; Hall Island, Behring Sea, coll. of 1885, Thompson (Field Mus. Herb.) ; Hall Island, Behring Sea, 11 Aug., 1891, Macoun 20640 (Geol. Surv. Canada Herb.) ; Herschel Island, Arctic Sea, coll. of 1893, Stringer 14390 (Geol. Surv. Canada Herb.); Port Clarence, 21 July, 1895, Sharp (Phil. Acad. Nat. Sci. Herb.) ; [Vor. 3 100 ANNALS OF THE MISSOURI BOTANICAL GARDEN Cape Nome, coll. of 1890, Blaisdell (Gray Herb.) ; vieinity of Nome, Powers 6 (Field Mus. Herb.) ; Cape Vancouver, 9 Aug., 1891, Macoun (Geol. Surv. Canada Herb., Gray Herb., and Mo. Bot. Gard. Herb.); Kuskokwim Valley, coll. of 1884, Weim- mann (Gray Herb.), branched form; Unga and Shumagin Islands, (7. 5. Coast Survey, 1871-72, Harrington (Gray Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.). Var. columbiensis Gray, Syn. Fl. N. Am. 17:390. 1884, and ed. 2, 1886; Macoun, Cat. Canadian РІ. 267. 1884. S. hyperborealis var. columbiensis (Gray) Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Ann. Mo. Bot. Gard. 1:264. 1914. Stems about 2 dm. high, one to several from a common base; heads subdiseoid, namely, with inconspicuous ray-flowers shorter than the bracts of the involucre., Distribution: British Columbia. Specimen examined : British Columbia: Mucklung River, 25 July, 1882, Mackay (Gray Herb.), TYPE. 40. 8. ovinus Greene, Pittonia 4:110. 1900; Blankenship, Mont. Agr. Coll. Sei. Studies 1:103. 1904. S. resedifolius Rydb. Mem. N. Y. Bot. Gard. 1:447. 1900, not Less. A low herbaceous perennial, glabrous or somewhat tawny, floccose-tomentulose in ће axils of the leaves; stems one to several from an ascending stoutish rootstock, .5 to 2 dm. high, erect or nearly so; lower leaves broadly ovate to obovate, occasionally sublyrate, including the petiole 1 to 6 em. long, the blade .5 to 3 em. long, .3 to 2.5 em. broad, subentire to erenate-dentate, glabrous, thickish in texture; upper leaves sessile, laciniate to entire; heads usually solitary, occasionally two, 8 to 10 mm. high, radiate; involucre companulate, spar- ingly ealyeulate; bracts of the involuere about 21, linear- lanceolate, acuminate, acute, 7 to 8 mm. long, often purplish; ray-flowers 13 to 21, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: mountains of Alberta to Wyoming. 1916] GREENMAN—MONOGRAPH OF SENECIO 101 Specimens examined: Alberta: high slopes of Sheep Mountain, Waterton Lake, 29 July, 1895, Macoun 11619 (Geol. Surv. Canada Herb., Gray Herb., photograph in Field Mus. Herb. and Mo. Bot. Gard. Herb.), TYPE. British Columbia: near the western summit of North Kootenai Pass, 26 July, 1883, Dawson (Geol. Surv. Canada Herb. 14826 in part, and Greene Herb.). | Montana: McDonalds Peak, Mission Range, alt. 2440 m., 19 July, 1883, Canby 36 (Gray Herb.); Stanton Lake, 1 Aug., 1894, Williams 1022 (Gray Herb., U. S. Nat. Herb., and Mont. Agr. Coll. Herb.); Mt. Hyalite, alt. 3000 m., 1 Aug., 1902, Blankenship (Gray Herb.); Sperry Glacier, 1 Sept. 1903, Umbach 798 (Field Mus. Herb.) ; Sperry Glacier, alt. 2440 m., 1 Sept., 1903, Blankenship (Gray Herb.); MacDougal Park, Flathead Lake and vicinity, 31 July, 1908, Clemens (Field Mus. Herb. and Mo. Bot. Gard. Herb.); Glacier National Park, 11 July, 1914, Hitchcock 11962 (U. S, Nat. Herb.). Wyoming: Wind River Mountains, alt. 2700-3000 m., “С.В.” (Gray Herb.). 41. 8. conterminus Greenm. nom. nov. S. Lyallii Klatt in Annal. Naturhist. Hofm. Wein 9:365. 1894, not Hook. f.; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. A low depressed somewhat cespitose herbaceous peren- nial, conspicuously white-floccose tomentose at the base and in the axils of the leaves; stems erect or nearly so, 4 to 8 cm. high from a stout rootstock; leaves thickish, the lower ovate- rotund to oblong-spatulate or occasionally sublyrate, crenate- dentate to obtusely lobed, including the petiole 1 to 3 em. long, 1 em. or less broad; stem-leaves sessile, laciniate, the upper- most reduced to entire purplish-tipped bracts; heads usually solitary, 10 to 14 mm. high, radiate; involucre campanulate, ealyeulate, floccose-tomentulose at the base, glabrous above; bracts of the involucre about 21, linear-lanceolate, acuminate, 8 to 10 mm. long, somewhat penicillate, and more or less tinged with purple; ray-flowers 10 to 12, rays yellow; disk-flowers numerous, slightly exceeding the involucre; achenes glabrous. [Vor. 3 102 ANNALS OF THE MISSOURI BOTANICAL GARDEN Distribution: Rocky Mountains, near the Canadian bound- ary, northward into Alberta and British Columbia. Specimens examined: Rocky Mountain summits, alt. 2130-2440 m., Oregon Bound- ary Commission, coll. of 1861, Lyall (Gray Herb., Kew Herb., and Berlin Herb.), түрк. Alberta: Sheep Mountain, Waterton Lake, 28-31 July, 1895, Macoun (Kew Herb. Berlin Herb. and U. S. Nat. Herb. 289213) ; on the summit of Moose Mountain, alt. 2285 m., 29 June-1l July, 1897, Macoun 22781, 22773 (Geol. Surv. Canada Herb.) ; on high mountain slopes, Crows Nest Pass, alt. 1825— 2289 m., 2 Aug., 1897, Macoun 22782, 22783 in part (Geol. Surv. Canada Herb.). British Columbia: north summit of North Kootenai Pass, eoll. of 26 July, 1883, Dawson (Geol. Surv. Canada Herb. 14826 in part). 42. 8. hesperius Greene, Pittonia 2:166. 1891. Р]. 3, fig. 1. S. hesperis Howell, Fl. Northwest Am. 1:375. 1900. S. pyroloides Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. S. auleticus Greene, Leafl. Bot. Obs. & Crit. 2:15. 1909. A slender herbaceous perennial; stem erect, 1 to 2 dm. high, floccose-tomentulose, especially at the base and in the axils of the leaves, later more or less glabrate, obovate-rotund to oblanceolate, including the petiole 1 to 6 em. long, .5 to 1.8 em. broad, entire to crenate-dentate, narrowed at the base into a petiole equalling or much exceeding the blade, at first tomentulose soon glabrate; stem-leaves sessile, laciniate to linear-attenuate; heads usually solitary, occasionally two, rarely five, 10 to 12 mm. high, radiate; involucre campanulate, sparingly calyculate, slightly tomentulose to glabrous; bracts of the involucre 13 to 21, linear-lanceolate, 5 to 8 mm. long; rays yellow; disk-flowers rather numerous; achenes glabrous. Distribution: southwestern Oregon. Specimens examined: . Oregon: Eight Dollar Mountain, May, 1884, Howell 160 (Gray Herb.) ; near Kirbyville, 27 May, 1884, Howell 1511 1916] GREEN MAN—MONOGRAPH OF SENECIO 103 (Greene Herb., U. S. Nat. Herb., Torrey Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb), түре; Eight Dollar Mountain, 13 June, 1904, Piper 6145 (U. S. Nat. Herb.) ; eight miles south of Waldo, 14 June, 1904, Piper 6254 (U. S. Nat. Herb.). 43. S. Newcombei Greene, Pittonia 3:249. 1897. A slender herbaceous perennial, glabrous throughout; stem solitary, erect or nearly so, 1.5 to 2 dm high; leaves thin, the lower petiolate, reniform to obovate in general outline, 3 to 7-lobate-dentate, 1 to 1.5 em. long, 1 to 2 em. broad, subeordate to abruptly narrowed at the base into a slender petiole, the lobes ovate-oblong, mucronate; upper leaves cuneate to linear; heads solitary, radiate, about 1 em. high; involucre subcam- panulate, ecalyculate, glabrous; bracts of the involucre 8 to 13, laneeolate, 6 to 8 mm. long, acute; ray-flowers 10 to 12, rays narrowly oblong, 1 to 1.5 em. long, 2 to 4 mm. broad, in the dried state more or less tinged with lilae; disk-flowers rather numerous; achenes striate, glabrous. Distribution: known only from Queen Charlotte Islands. Specimens examined: British Columbia: Kaitgoro, west coast of A m Island, Queen Charlotte Islands, 28 June, 1897, Newcombe (Greene Herb., Geol. Surv. Canada Herb. 18707, and Kew Herb., trac- ing in Gray Herb.), түре; in the same locality, coll. of 1903, Newcombe (Field Mus. Herb. and Gray Herb.). This species is known only from its original station, and while it exhibits certain characteristics not common in Senecio, yet it has the technical floral characters of this genus. 44. S. subnudus DC. Prodr. 6:428. 1837; Nutt. in Trans. Am. Phil. Soe, 7:412. 1841; Torr. & Gray, Fl. N. Am. 2:445. 1843; Howell, Fl. Northwest Am. 378. 1900; Rydb. Mem. N. Y. Bot. Gard. 1:447. 1900, and Bull. Torr. Bot. Club 27:184. 1900; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Piper, Contr. U. S. Nat. Herb. 11:597. 1906; Coulter & Nelson, Manual Cent. Rocky Mountains 582. 1909. S. aureus var. subnudus Gray, Syn. Fl. N. Am. 1?:391. 1884, and ed. 2, 1886; Macoun, Cat. Canadian Pl. 266. 1884. [Vor. 3 104 ANNALS OF THE MISSOURI BOTANICAL GARDEN S. cymbalarioides Buek, Ind. DC. Prodr. pt. 2, p. vi. 1840, not Nutt. A slender herbaceous perennial, glabrous throughout; stem simple, erect, 1 to 3.5 dm. high, rising from a slender root- stock; leaves thin, membranous, the lower petiolate, sub- rotund-obovate, occasionally sublyrate, including the petiole .9 to 9 em. long, .5 to 3 em. broad, abruptly to gradually nar- rowed at the base, crenate-dentate; the upper leaves sessile, incised to entire; heads solitary or occasionally two, 8 to 10 mm. high, including the yellow rays 1.5 to 2.5 em. in diameter ; bracts of the involucre about 21, linear-lanceolate, acute, glabrous and often purplish, slightly shorter than the rather numerous flowers of the disk; achenes glabrous. Distribution: Montana and Wyoming to Washington and California. Specimens examined; Montana: bogs, Park Co., alt. 2800 m., Aug., 1897, Tweedy 344 (Mont. Agr. Coll. Herb.); in open bogs among the hills, West De Lacy Creek, 4 Aug., 1899, А. Ё E. Nelson 6300 (Gray Herb. U. S. Nat. Herb, and Mo. Bot. Gard. Herb.); Old Hollowtop, near Pony, alt. 2740 m., 7 July, 1897, Rydberg & Bessey 5270 (Gray Herb., U. S. Nat. Herb., Field Mus. Herb., and Mont. Agr. Coll. Herb.). Yellowstone National Park: Upper Falls of the Yellow- stone, Hayden's U. S. Geol. Survey, 1871, Adams (U. S. Nat. Herb.) ; in bogs, alt. 2740 m., Aug., 1885, Tweedy 720 (U.S. Nat. Herb. 143107, and Field Mus. Herb.). Rocky Mountains: Lat. 49° N., alt. 1980 m., Oregon Bound- ary Commission, 1861, Lyall (Gray Herb.). Wyoming: without definite locality or date of collection, Tweedy 585 (U. S. Nat. Herb. 48711); Wind River Chain of Rocky Mountains, alt. 2135 m., Fremont (Gray Herb.). Idaho: ridges south from Wiessner’s Peak, alt. 1900 m., 27 July, 1895, Leiberg 1376 (С. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Bear Creek Cañon, alt. 1500 m., 1 Sept., 1897, Leiberg 2968 (U. S. Nat. Herb.). Washington: Horse Shoe Basin, Okanogan Co. Sept., 1897, Elmer 1423 (Mo. Bot. Gard. Herb.); Yakima region,. 1916] GREENMAN—MONOGRAPH OF SENECIO 105 Northern Transcontinental Survey, coll. of 1882, Brandegee 118 (Mo. Bot. Gard. Herb.) and 915 (Gray Herb.); wet meadows, Chiquash Mountains, Skamania Co., 18 Aug., 1892, Suksdorf 2167 (Gray Herb. U. S. Nat. Herb, Field Mus. Herb., and Mo. Bot. Gard. Herb.) ; north of Mt. Adams, Aug., 1892, Henderson 2308 (Gray Herb.) ; wet meadows, alt. about 2000 m., Mt. Paddo (Adams), 3 Sept., 1904, S'uksdorf 4241 (U. S. Nat. Herb., Field Mus. Herb, and Mo. Bot. Gard. Herb.). Oregon: Eagle Creek Mountains, Union Co., alt. 2440 m., Aug., 1881, Cusick 938, 959 (Gray Herb.) ; wet meadows of the highest mountains, eastern Oregon, coll. of 1897, Cusick 1804 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; granitic soil, swampy meadows, North Catherin Creek, eastern Oregon, alt. 1500 m., 26 July, 1907, Cusick 3177 (U. S. Nat. Herb., Field Mus. Herb, and Mo. Bot. Gard. Herb.); sedgy meadow, Wallowa Mountains, source of Kettle Creek, alt. 2285 m., 12 Aug., 1909, Cusick 3379 (U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.) ; ‘Cascade Mountains on the Ore- gon” (Douglas?) Dr. Gairdner (Gray Herb.); without lo- eality, coll. of 1871, Hall 304 (Gray Herb. and Mo. Bot. Gard. Herb.) ; base of Mt. Hood, June, 1878, J. Ё S. J. Howell (Field Mus. Herb.) ; Mt. Hood, alt. 1220 m., 25-26 Aug., 1914, Hitch- cock 12328 (U. S. Nat. Herb.) ; Caseade Mountains, Aug., 1880, Howell (U. S. Nat. Herb. 48811, and Field Mus. Herb.). California: Plumas Co., coll. of 1876, Mrs. Austin (Gray Herb. and Field Mus. Herb.); summit of Mt. Dana, coll. of 1878, Lemmon (Gray Herb.); Pine Creek, July, 1912, Baker (Field Mus. Herb.). 45. S. Rosei Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Field Col. Mus. Bot. Ser. 2:276. 1907. РІ. 3, fig. 2. An herbaceous perennial; stem solitary, erect, 4 to 4.5 dm. high, simple, glabrous below, slightly pubescent above, ter- minated by a single large radiate head ; lower leaves petiolate, ovate, 2 to 3 em. long, two-thirds as broad, obtuse, subentire to crenate-dentate, thin, glabrous on both surfaces; petioles [Vor. 3 106 ANNALS OF THE MISSOURI BOTANICAL GARDEN 2.5 to 8 em. long; upper stem-leaves sublaciniate and some- what ampliated at the base and partly clasping the stem; heads about 12 mm. high, including the rays 3 to 3.5 em. in diameter; involucre campanulate, essentially ecalyculate; bracts of the involucre lanceolate-linear, 8 to 10 mm. long, acute, glabrous; ray-flowers 10 to 12, rays light yellow, con- spicuous; disk-flowers numerous; achenes glabrous. Distribution: west central Mexico. Specimen examined: Territory of Tepic: Sierra Madre, near Santa Teresa, 10 Aug., 1897, Rose 2157 (Gray Herb. and U. 8. Nat. Herb.), TYPE. 46. S. Porteri Greene, Pittonia 3:186. 1897; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Rydb. КІ. Colo. 397. 1906. S. renifolius Porter in Porter & Coulter, Syn. Fl. Colo. 83. 1874, not Schz. Bip.; Gray, Syn. Fl. N. Am. 1:389. 1884, and ed. 2, 1886; Coulter, Manual Rocky Mountain Region 210. 1885. | A low herbaceous perennial, glabrous throughout and more or less tinged with purple; stems ascending from a slender rootstock, 1 dm. or less high; leaves petiolate, mostly reniform, including the petiole 1.5 to 5 em. long, .8 to 2.5 cm. broad, erenate; heads about 14 mm. high, solitary on nearly naked peduncles, radiate; involucre campanulate, sparingly ealycu- late; bracts of the involucre about 13, linear-lanceolate, acute, 10 mm. long, purplish; ray-flowers 8 to 10, rays bright yellow; disk-flowers numerous; achenes glabrous. Distribution: high mountains of Colorado and eastern Oregon. Specimens examined: Colorado: White House Mountain, alt. 3960 m., Hayden’s U. S. Geol. Survey, 9 Aug., 1873, Coulter 2950 (Gray Herb., Phil. Acad. Nat. Sci. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.). Oregon: alpine ridges of the Wallowa Mountains, 3 Aug., 1899, Cusick 2308 (Gray Herb., Kew Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.). 1916] GREENMAN—MONOGRAPH OF SENECIO 107 47. 8. Soldanella Gray, Proc. Acad. Nat. Sci. Phil. 15:67. 1863; Porter & Coulter, Syn. Fl. Colo. 83. 1874; Gray, Syn. КІ. N. Am. 1?:384. 1884, and ed. 2, 1886; Coulter, Manual Rocky Mountain Region 206. 1885; Rydb. Fl. Colo. 394. 1906; Coul- ter & Nelson, Manual Cent. Rocky Mountains 578. 1909; Cle- ments & Clements, Rocky Mountain Fls. 291. 1914. S. Grayi Parry ex Gray, Proc. Acad. Nat. Sci. Phil. 15:67. 1863, not S. Greyi Hook. f. A low herbaceous perennial, 1 to 2 dm. high, glabrous throughout and more or less tinged with purple; stems flex- uous, ascending from a stoutish rootstock, the latter bearing numerous fleshy-fibrous roots; leaves somewhat suceulent, subrotund to oblong-obovate, 1.5 to 5.5 em. broad, entire to sinuate-dentate, subeordate or more frequently euneate at the base into a winged petiole much exceeding the blade; heads large, 1.5 to 2 em. high, usually solitary, radiate; involucre broadly campanulate, calyculate; bracts of the involucre narrowly lanceolate, 10 to 14 mm. long, pubescent-tipped, otherwise glabrous and purplish; ray-flowers 10 to 18, rays yellow; disk-flowers very numerous; achenes strongly ribbed, glabrous. Distribution: high mountains of Colorado. Specimens examined: Colorado: Gray's Peak, 1 Aug., 1862, Parry (Gray Herb., Kew Herb., and Mo. Bot. Gard. Herb.), түре; Lat. 39°41’ N., coll. of 1862, Най € Harbour 319 (Gray Herb., Kew Herb., U.S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.) ; South Park, alt. 3960 m., Lieut. Wheeler’s Expedition, 1873, . Wolf £ Rothrock 573, 575 (Gray Herb. and U. S. Nat. Herb.) ; Mt. La Plata, alt. 4265 m., Hayden's U. 8. Geol. Survey, 1873, Coulter (Gray Herb. and Field Mus. Herb.) ; Sangre de Cristo, Aug., 1873, Brandegee 724 (U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.) ; Sawatche Range, coll. of 1880, Brandegee (Mo. Bot. Gard. Herb.) ; high mountains, Gray's Peak and vicinity, alt. 3350-4265 m., Aug., 1885, Pat- terson 84 (Gray Herb., Kew Herb., U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.); Gray's Peak, alt. . [Vor. 8 108 ANNALS OF THE MISSOURI BOTANICAL GARDEN 4965 m., 21 July, 1886, Letterman 67 (Mo. Bot. Gard. Herb.) ; Стау в Peak, July, 1888, Eastwood (U. 5. Nat. Herb.); Mt. Baldy, alt. 3655 m., 11 July, 1891, Smith (Mo. Bot. Gard. Herb.) ; Mt. Princeton, alt. 2115 m., 21 July, 1892, Sheldon 172, 491 (U. S. Nat. Herb.) ; stony slopes of Sheep Mountain, alt. 4995 m., coll. of July, 1893, Purpus 680 (Field Mus. Herb.) ; La Plata Mountains, alt. 3350-3600 m., 15 July, 1896, Tweedy 536 (U. S. Nat. Herb.) ; Cameron Pass, alt. 3600 m., 16 July, 1896, Baker (Mo. Bot. Gard. Herb.); Cumberland Mine, La Plata Mountains, alt. 3750 m., 15 July, 1898, Baker, Earle & Tracy 534 (Gray Herb, Kew Herb., U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.); Mt. Baldy, near Breckenridge, alt. 3960 m., Aug., 1901, Mackenzie 164 (Phil. Acad. Nat. Sci. Herb. and Mo. Bot. Gard. Herb.). 48. S. obovatus Muhl. ex.Willd. Sp. Pl. 3:1999. 1804; Pursh, Fl. Am. Sept. 2:530. 1814, and ed. 2, 1816; Nutt. Gen. 2:165. 1818; Ell. Sketch 329. 1824; Eaton, Manual of Botany 454. 1824; DC. Prodr. 6:432. 1837; Heller, Cat. N. Am. PL 146. 1898, and ed. 2, 230. 1900; Britton & Brown, Ill. Fl. 3:478, fig. 4041. 1898, and ed. 2, 545, fig. 4627. 1913; Britton, Manual 1027. 1901, and ed. 2, 1905; Greenm. Rhodora 3:5. 1901; Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Porter, Fl. Penn. 339. 1903; Keller & Brown, Handb. Fl. Phil. and Vicinity 343. 1905; Small, Fl. South- eastern United States 1304. 1903, and ed. 2, 1913; Greenm. in Gray, Manual, ed. 7, 854. 1908; Graves et al. Conn. Geol. and Nat. Hist. Surv. Bull. No. 14, p. 404. 1910; Small & Carter, Fl. Laneaster County 310. 1913. S. obtusatus Banks ex Pursh, Fl. Am. Sept. 2:530. 1814, and ed. 2, 1816; DC. Prodr. 6:432. 1837. S. Elliottii Torr. & Gray, Fl. N. Am. 2:443. 1843; Chapman, Fl. Southern U. S., ed. 1, 245. 1860, and ed. 2, 1889. S. aureus var. obovatus Torr. & Gray, Fl. N. Am. 2:442. 1843; Gray, Syn. Fl. N. Am. 12:391. 1884, and ed. 2, 1886; Macoun, Cat. Canadian РІ. 265. 1884, in part; Chapman, КІ. Southern U. S., ed. 3, 266. 1897. An herbaceous perennial, glabrous or slightly tomentulose ; stems erect, 2 to 5 dm. high, simple or branched, often stoloni- 1916] GREENMAN—-MONOGRAPH OF SENECIO 109 ferous at the base; stolons slender and elongated to short and rather stout; lower leaves petiolate, obovate, oblong-ovate to subrotund, 1 to 10 em. long, two-thirds to nearly or quite as broad, rounded at the apex, erenate to doubly serrate, grad- ually narrowed at the base and decurrent on the petiole or abruptly contraeted to a subeordate base; stem-leaves petio- late and sublyrate to sessile, more or less pinnatisect and semiamplexicaul; inflorescence a terminal several to many- headed corymbose cyme; heads 8 to 10 mm. high, radiate; ray- flowers 8 to 12, rays relatively long and narrow, yellow; disk- flowers numerous; achenes glabrous or occasionally hispidu- lous along the angles. Distribution: Vermont, south to Georgia, west to Mis- souri, Kansas, and Texas. Specimens examined: Vermont: Pownal, Bennington Co., 31 July, 1898, Eggles- ton 264 (Gray Herb.), and 8-11 Sept., 1899, Eggleston 1381 (U. S. Nat. Herb.). Massachusetts: Ipswich, coll. of 1842, Oakes (Gray Herb. and U. S. Nat. Herb.); Boxford, 22 June, 1878, E. Faxon (Gray Herb.) ; Alford, Milligan (U. S. Nat. Herb.) ; without definite locality, Nuttall (Phil. Acad. Nat. Sci. Herb.). Connecticut: shaded limestone ledges, Salisbury, 3 June, 1901, Bissell (Gray Herb.) ; in loam and on calcareous ledges, Salisbury, 30 May, 1902, Churchill, Bissell & Fernald 96 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.) ; near Southington, 24 Aug., 1903, Andrews (Gray Herb.) ; Meriden Mountain, 30 May, 1885, Safford 344 (U. S. Nat. Herb.) ; rich dry rocky woods, Monroe Co, 8 June, 1895, Fames (U. S. Nat. Herb.) ; thin dry woods on ledge of trap rock, Long Hill, 26 May, 1894, Eames (Gray Herb.); Bridgeport, 19 May, 1893, Eames (U. S. Nat. Herb.). New York: Lebanon Springs, 31 May, 1890, Harrison 9 (U. S. Nat. Herb.); West Point, er Herb. Thurber (Gray Herb.) ; Van Cortlandt, May, 1893, Pollard (U. S. Nat. Herb.) ; Delaware Co., 6 July, 1892, H. von Schrenk (Mo. Bot. Gard. Herb.); without definite locality, Torr. & Gray, Fl. N. Am. (Gray Herb.). à [VoL. 8 110 ANNALS OF THE MISSOURI BOTANICAL GARDEN New Jersey: dry soil, Cranberry Lake, Sussex Co., 9 June, 1907, Mackenzie 2616 (U. 8. Nat. Herb.); Budd’s Lake, Morris Co., 29 May, 1895, Heritage, in part, and 30 May, 1899, Lippincott, in part (Phil. Acad. Nat. Sci. Herb.) ; rocky banks, near Charlotteburg, Morris Co., 24 May, 1908, Mackenzie 3089 (Mo. Bot. Gard. Herb.) ; above Phillipsburg, Warren Co., 18 May, 1907, Van Pelt & Long (Phil. Acad. Nat. Sci. Herb.). Pennsylvania: Stroudsburg, 2 June, 1900, ez Herb. Canby (Phil. Acad. Nat. Sci. Herb.) ; Bartonsville, 1 July, 1907, Long Ф Bartram (Phil. Acad. Nat. Sci. Herb.); side of Pocono Knob, 31 May, 1902, Brown (Phil. Acad Nat. Sci. Herb.) ; Easton, Trail Green (Gray Herb.) ; Buckskill Falls, 31 May, 1897, Brown (Phil. Acad. Nat. Sci. Herb.) ; Hellertown, 1 June, 1849, ex Herb. Detwiller (Phil. Acad. Nat. Sci. Herb.) ; along Jordan Creek, Lehigh Co., 14 May, 1911, Pretz 3336 (Phil. Acad. Nat. Sci. Herb.) ; Nockamixon, 28 May, 1893, MacElwee, in part, 19 May, 1906, Van Pelt & Long, 26 Мау, 1902, Fretz, and 4 June, 1894, Crawford (Phil. Acad. Nat. Sci. Herb.) ; Nockamixon Rocks, 30 May, 1893, N. L. Britton (U. S. Nat. Herb.) ; Narrowsville, 30 May, 1893, Brown (Phil. Acad. Nat. Sci. Herb.) ; Ivy Roek, below Norristown, 9 May, 1896, Keller, and 6 Мау, 1906, Van Pelt (Phil. Acad. Nat. Sci. Herb.) ; Pennsburg, 20 June, 1910, Mumbauer 255 (Phil. Acad. Nat. Sci. Herb.) ; Arcola, ex Herb. Crawford (Phil. Acad. Nat. Sci. Herb.); Phoenixville, June, 1865, ex Herb. Martindale, and 25 April, 1909, Bartram (Phil. Acad. Nat. Sci. Herb.) ; near Philadelphia, Griffith 30 (Phil. Acad. Nat. Sci. Herb.) ; Phil- adelphia, ex Herb. Bernhardi (Mo. Bot. Gard. Herb.) ; Sehuyl- kill, 5 May, 1834, collector not indicated (Phil. Acad. Nat. Sci. Herb.); Lafayette and Sumneytown, Williamson (С. 8. Williamson Herb.) ; in limestone on the Conestoga near Dan- ville Pike Crossing, 17 May, 1901, Heller (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Conestoga, April, 1889, Eby (Mo. Bot. Gard. Herb.); Mountville, Laneaster Co., April, 1889 and 1890, Eby (Mo. Bot. Gard. Herb.) ; Me- Connellsburg, Fulton Co., 3 June, 1905, Stone 234 (Phil. Acad. Nat. Sei. Herb.); Johnstown, Cambria Co., Williams (C. S. 1916] GREENMAN—-MONOGRAPH OF SENECIO TI Williamson Herb.), and 16 June, 1907, Bartram (Phil. Acad. Nat. Sci. Herb.) ; Homewood, 15 June, 1907, Bartram (Phil. Acad. Nat. Sci. Herb.) ; Ohiopyle, Fayette Co., 3-8 July, 1905, Brown, Crawford & Van Pelt 22, 135 (Phil. Acad. Nat. Sci. Herb.). Virginia: Great Falls of the Potomae, 21 April, 1908, Wil- liamson (Phil. Acad. Nat. Sci. Herb.) ; on limestone cliffs at edge of river near Schulers, west of Luray, Page Co., 10 May, 1904, G. S. Miller (U. S. Nat. Herb.) ; about Mt. Crawford, Rockingham Co., alt. 365-455 m., 5-13 May, 1893, Heller (С. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.); bluffs of Middle Fork of Holston River, near Marion, Smyth Co., alt. 640 m., 22 May, 1892, Small (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; vicinity of Marion, 22 May, 1892, N. L. and E. С. Britton Ф Vail (Phil. Acad. Nat. Sci. Herb.). North Carolina: rocky woods, Hewitt's Station, Swain Co., 24 May, 1897, Biltmore Herb. 3364a (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Hot Springs and Warm Springs, 24 April, 1887, Aubrey H. Smith (Phil. Acad. Nat. Sci. Herb.). South Carolina: slopes of Paris Mountain, Greenville, 2 April, 1908, Mackenzie (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.), tomentulose form. Georgia: bluffs, Rome, coll. of 1882, ex Herb. Chapman (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.), a form approaching the variety rotundus. Alabama: Boykin, Buckley (Gray Herb.); Moulton, Law- rence Co., Mohr (U. S. Nat. Herb.) ; without definite locality, Wilkinson (U. S. Nat. Herb.). West Virginia: White Sulphur Springs, 17-18 May, 1909, Eggleston 4346 (Mo. Bot. Gard. Herb.) ; Fairmont, May, 1907, Williamson (C. S. Williamson Herb.). Ohio: near Canton, May, 1875, ex Herb. Riddell 149 (Mo. Bot. Gard. Herb.); river banks, Lorain Co., 21 April, 1890, Kofoid (Mo. Bot. Gard. Herb.) ; Huron River, Erie Co., Mose- ley (Gray Herb.) ; Oxford, 2 June, 1908, Overholts (Mo. Bot. Gard. Herb.). [Vor. 3 112 ANNALS OF THE MISSOURI BOTANICAL GARDEN Indiana: around lakes, Wells Co., 11 May, 1908, Deam (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; river bank, north of Notre Dame, 6 June, 1911, Niewwland 2660, and 15 May, 1913, Nieuwland 11114 (Mo. Bot. Gard. Herb.) ; Worth- ington, 29 April, 1889, Evermann (U. S. Nat. Herb.). Kentucky: beargrass hills near Louisville, 20 May, 1853, Mohr (U. S. Nat. Herb.) ; without definite locality, ex Herb. Short (Phil. Acad. Nat. Sci. Herb.). Tennessee: rich bluffs, Knoxville, May, 1896, Ruth (2) and in woods, near Knoxville, June, 1896, Ruth 5 (Mo. Bot. Gard. Herb.) ; Knoxville, 5 May, 1895, J. G. Smith (Mo. Bot. Gard. Herb.); moist woods, Cumberland Mountain, 5 May, 1898, Eggert (Mo. Bot. Gard. Herb.) ; cedar glades, near La- vergne, 4 May, 1898, Eggert (Mo. Bot. Gard. Herb.). Missouri: Jerome, colls. of 1913 and 1914, J. H. Kellogg 56, 58, 90, 452 (Mo. Bot. Gard. Herb.) ; Swan, 21 April, 1907, Bush 4203 (Mo. Bot. Gard. Herb.) ; Monteer, 12 May, 1901, Bush 429, and 14 Мау, 1905, Bush 2836 (Mo. Bot. Gard. Herb.) ; on bluffs, Neck City, Jasper Co., 12 May, 1912, Е. J. Palmer 3669 (Mo. Bot. Gard. Herb.). Arkansas: Independence, 23 April, 1896, Eggert (Mo. Bot. Gard. Herb.) ; Eureka Springs, 20 April, 1899, Trelease (Mo. Bot. Gard. Herb.); Hartford Station, April, 1903, Pilsbury (Phil. Acad. Nat. Sci. Herb.); Petit Jean, Yell Co., 1 and 2 April, 1903, Pilsbury (Phil. Acad. Nat. Sei. Herb.). Oklahoma: Vinita, 23 April, 1891, Carleton 19 (U. S. Nat. Herb.); on creek bank, Page, Leflore Co., 20 April, 1915, Blakeley 3443 (Mo. Bot. Gard. Herb.). Texas: river banks, Denison, 28 March, 1890, Bodin 27 (U. S. Nat. Herb.) ; near Austin, 17 March, 1908, York (Mo. Bot. Gard. Herb.). Var. divisifolius Greenm. var. nov. Stems bearing short stout stolons; lower leaves obovate to narrowly oblong-oblanceolate, those of the stem relatively long and conspicuously pinnatisect. A local but striking variation from the type. Distribution: bluffs near Knoxville, Tenn. 1916] GREENMAN—MONOGRAPH OF SENECIO 113 Specimens examined : Tennessee: rich woods, Knoxville, April, 1898, Ruth 705 (Mo. Bot. Gard. Herb.), түре; bluffs, Knoxville, April, 1898, Ruth 674 (U. S. Nat. Herb.) ; bluffs, Knoxville, May, 1896, Ruth (3) (Mo. Bot. Gard. Herb.); vicinity of Knoxville, 29 April, 1890, Lamson-Scribner (U. S. Nat. Herb.). Var. elongatus (Pursh) Britt. in Britton & Brown, Ill. КІ. 3:478. 1898; Britton, Manual 1027. 1901, and ed. 2, 1905; Porter, Fl. Penn. 339. 1903; Keller & Brown, Handb. Fl. Phil. and Vicinity 343. 1905; Greenm. Monogr. Senecio, I. Teil, 24. 1901, in Engl. Bot. Jahrb. 32:20. 1902, and in Gray, Manual, ed. 7, 854. 1908. S. elongatus Pursh, Fl. Am. Sept. 2:529. 1814, and ed. 2, 1816. S. aureus var. discoidea Porter in herb., not S. aureus var. discoideus Hook. Habit and foliage of the species; peduncles of the inflo- rescence relatively long; heads discoid. Distribution: eastern Pennsylvania and New Jersey. Specimens examined: New Jersey: Budd’s Lake, Morris Co., 29 May, 1895, Heritage, in part, and 30 May, 1895, Lippincott, in part (Phil. Acad. Nat. Sci. Herb.). Pennsylvania: vicinity of Easton, coll. of 1807, Pursh, ex Herb. Lambert (Gray Herb.); College Hill, Easton, June, 1867, Porter (Gray Herb.) ; College Hill, 1 June, 1868, Porter, and coll. of 1868, Garber (Phil. Acad. Nat. Sci. Herb.) ; Col- lege Hill, June, 1870, Porter (U. S. Nat. Herb.) ; College Hill, May, 1871, Porter (Mo. Bot. Gard. Herb.); Spruce Hill on Bushkill Creek, 25 May, 1887, Porter (Phil. Acad. Nat. Sci. Herb.); College Hill, 12 May, 1890, Porter (Gray Herb. and О. S. Nat. Herb.) ; Мейр, Bueks Co., 28 May, 1898, MacElwee (Phil. Acad. Nat. Sci. Herb.), in natt; College Hill, 17 May, 1895, Porter (Phil. Acad. Nat. Sci. Herb.) ; limestone bluffs on the Bushkill, 5 May, 1899, Porter (Phil. Acad. Nat. Sci. Herb.) ; “hillsides, near the Schuylkill and Susquehanna ’’, ex. Herb. David Townsend (Phil. Acad. Nat. Sci. Herb.). 8 [Vor. 3 114 ANNALS OF THE MISSOURI BOTANICAL GARDEN Var. rotundus Britt. in Britton & Brown, Ill. Fl. 3:479. 1898. Britton, Manual 1027. 1901, and ed. 2, 1905; Greenm. Monogr. Senecio, I. Teil, 24. 1901, in Engl. Bot. Jahrb. 32:20. 1902, and in Gray, Manual 854. 1908; Blankenship in Mo. Bot. Gard. Ann. Rept. 18:179. 1907. S. rotundus Small, Fl. Southeastern U. S. 1304. 1903, and ed. 2, 1913. S. Lindheimeri Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. Habit, inflorescence, and technical characters of the head like the species; basal leaves usually long-petioled, the blade obovate to subrotund, cuneate to abruptly constricted into a subeordate base. Distribution: Ohio to eastern Kansas and south to Louisi- ana and Texas. Specimens examined : Ohio: Oak Point, Lorain Co., 11 Мау, 1895, Ricksecker (U. S. Nat. Herb.) ; Franklin Co., 7 May, 1892, Werner 133 (Gray Herb.) ; central Ohio, Sullivant (Gray Herb.) ; vieinity of Miami, coll. of 1835, Dr. Frank (Mo. Bot. Gard. Herb.) ; Dayton, collector not indicated (Gray Herb.) ; near Cincinnati, Lloyd (Mo. Bot. Gard. Herb.). Missouri: flood plain of Fox Creek, near Allenton, St. Louis Co., 25 May, 1914, Drushel (Mo. Bot. Gard. Herb. and J. A. Drushel Herb.) ; wet places near Glencoe, 22 May, 1879, Eg- gert (Mo. Bot. Gard. Herb.); Meramee Highlands, 7 May, 1898, Norton (Mo. Bot. Gard. Herb.) ; Pacifie, 24 May, 1915, Drushel (J. A. Drushel Herb.); Jefferson City, 6 May, 1866, Krause (Mo. Bot. Gard. Herb.); Kimmswiek, 10 May, 1885, Wislizenus 226 (Mo. Bot. Gard. Herb.), in part; Monteer, 11 May, 1905, Bush 2822 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Pleasant Grove, 20 May, 1900, Bush 340, 710 (Mo. Bot. Gard. Herb. and U. S. Nat. Herb.) ; near Webb. City, 4 Мау, 1902, E. J. Palmer 314 (Mo. Bot. Gard. Herb.) ; Oronogo, 13 and 30 May, 1906, E. J. Palmer 859 (Mo. Bot. Gard. Herb.); La Russell, 22 May, 1908, E. J. Palmer 1342 (Mo. Bot. Gard. Herb.) ; Prosperity, 11 May, 1909, E. J. Pal- 1916] GREENMAN—MONOGRAPH OF SENECIO 115 mer 1667 (Mo. Bot. Gard. Herb.) ; Alba, 30 Мау, 1909, E. J. Palmer 1834 (Mo. Bot. Gard. Herb.) ; Carterville, 14 April, 1912, E. J. Palmer 3521 (Mo. Bot. Gard. Herb.) ; Eagle Rock, 22 May, 1898, Bush 262 (Mo. Bot. Gard. Herb.) ; open ground, McDonald Co., 24 May, 1891, Bush 871 (Mo. Bot. Gard. Herb.) ; rich hillside woods, Noel, 4 May, 1914, E J. Palmer 5478 (Mo. Bot. Gard. Herb.). Arkansas: Fulton, 18 April, 1901, Sargent, Trelease & Bush 143 (Phil. Acad. Nat. Sci. Herb.) ; Texarkana, 6 April, 1905, Bush 2241 (Mo. Bot. Gard. Herb.). Louisiana: without definite locality, Hale (Gray Herb.). Kansas: wet soil, Bourbon Co., 5 May, 1897, Hitchcock 1118 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.). Oklahoma: Sapulpa, 29 April, 1895, Bush 976 (Mo. Bot. Gard. Herb.). Texas: San Augustine, Crocket (U. S. Nat. Herb. 500119) ; Livingston, Polk Co., 9 April, 1914, E. J. Palmer 5189 (Mo. Bot. Gard. Herb.) ; Grand Saline, Reverchon (Mo. Bot. Gard. Herb.) ; Marshall, Harrison Co., 17 April, 1914, E. J. Palmer 5285 (Mo. Bot. Gard. Herb.) ; Mill Creek bottom, Washington Co., 26 Feb., 1844, Lindheimer (Mo. Bot. Gard. Herb. and Gray Herb.) ; rich woods, Dallas, 13 March, 1901, Reverchon 556 (Mo. Bot. Gard. Herb.) ; Gillespie Co., Jermy (Mo. Bot. Gard. Herb.) ; Comal Spring, New Braunfels, Lindheimer 446 (Mo. Bot. Gard. Herb.) ; New Braunfels, соПв. of 1850 and 1851, Lindheimer 510, 958, 959, 960 (U. S. Nat. Herb., Phil. Acad. Nat. Sei. Herb, and Mo. Bot. Gard. Herb); New Braunfels, alt. 228 m., 17-19 April, 1903, Pilsbury (Phil. Acad. Nat. Sci. Herb.); without definite locality, Wright (Gray Herb.). Var. umbratilis Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1909. Lower leaves petiolate, obovate, oblong-ovate to oblong- elliptie, 2 to 8 em. long, 1.5 to 5.5 em. broad, petioles 2 to 12 em. long. Distribution: occurring occasionally with the species, especially in shady plaees from Indiana and Virginia to Louisiana. [Vor. 3 116 ANNALS OF THE MISSOURI BOTANICAL GARDEN Specimens examined : Indiana: without definite locality, Clapp (Gray Herb.), TYPE. Virginia: shaded rocks, Bedford Co., 9 May, 1871, A. H. Curtiss (Gray Herb.). Kentueky: flat wet barrens, Henderson Co., 5 May, 1842, Short (Phil. Acad. Nat. Sci. Herb.). Tennessee: marsh, Jackson, 15 April, 1893, Bain 421 (Gray Herb.). Missouri: Monteer, 27 April, 1907, Bush 4337 (Mo. Bot. Gard. Herb.). Arkansas: in woods, Fulton, 15 April, 1902, Bush 1356 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; on low ground, Fulton, 17 April, 1905, Bush 2355 (Mo. Bot. Gard. Herb.) ; Judsonia, 15 May, 1877, Reynolds (Field Mus. Herb.) ; Texarkana, 16 May, 1901, T'release (Mo. Bot. Gard. Herb.) ; without definite locality, Dr. Pitcher (Phil. Acad. Nat. Sci. Herb.). 49. S. Cardamine Greene, Bull. Torr. Bot. Club 8:98. 1881; Gray, Syn. Fl. N. Am. 1? :390. 1884, and ed. 2, 1886; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Wooton & Standley, Contr. U. S. Nat. Herb. 19:747. 1915. A low herbaceous perennial, glabrous throughout or slightly tomentulose in the leaf-axils; roots fibrous; stems one to sev- eral from a stoutish rootstock, 1 to 3.5 dm. high; leaves mostly radical, petiolate, round-ovate, 1 to 5 em. long and broad, deeply eordate, erenate, green above, purple-tinged beneath; petioles 2 to 9 em. long; stem-leaves 1 to 3, more or less am- plexicaul, the uppermost sessile and much reduced; heads few, 1 to 3, about 1 em. high, radiate; involucre campanulate, spar- ingly ealyculate; bracts of the involucre lanceolate, 6 to 8 mm. long; ray-flowers 8 to 10, rays yellow; disk-flowers numerous, achenes glabrous. Distribution: mountains of southwestern New Mexico. Specimens examined: 1916] GREENMAN—MONOGRAPH OF SENECIO 117 New Mexico: Mogollon Mountains, 25 April, 1881, Greene (Gray Herb., Greene Herb., Kew Herb., Torrey Ны; Field Mus. Herb., KS Mo. Bot. dani Herb.), түре; Mogollon Mou tains, Willow Creek, 8 Aug., 1900, Wooton (U. S. Nat. Herb. 139065). 50. Ж cyclophyllus Greenm. Field Col. Mus. Bot. Ser. 2:276. 1907. An ка perennial; stem simple, 3.5 to 5 dm. high, sparingly tawny tomentulose at the base and in the axils of the leaves, otherwise glabrous, striate, somewhat purplish; radical and lowermost stem-leaves subrotund, 4 to 7 cm. long, equally broad, deeply cordate, crenate-dentate, green and glabrous above, purple beneath, petioles 5 to 8 em. long; upper stem-leaves sessile, amplexicaul, pinnately divided into narrowly obovate-cuneate, unequally dentate lateral divisions and a relatively large subreniform terminal division; inflo- rescence a terminal many-headed subeorymbose суше; heads 7 to 9 mm. high, radiate; involucre campanulate, sparingly ealyeulate with minute bracteoles, glabrous; bracts of the involucre about 21, lance-linear, 5 to 6 mm. long, acute, more or less purple-tipped; ray-flowers about 13, rays yellow; disk- flowers 50 to 60; mature achenes 2 mm. long, hispidulous. Distribution: northeastern Mexico. Specimens examine Nuevo Leon: near — coll. of 1906, Pringle 10230 (Gray Herb., photograph in Field Mus. Herb. and Mo. Bot. Gard. Herb.), type; Cerro la Scilla, near Monterey, 20 March, 1902, Nelson 6672 (Gray Herb.). 51. S. quebradensis Greenm.! 'Senecio quebradensis Greenm. sp. nov., herbaceus perennis glabrus vel praesertim in axillis foliorum albo-floccosus ; caule simpliee vel ramoso 1.5-4 dm alto да glabro; foliis inferioribus petiolati is rotundo-ovatis vel oblongo- -ovatis mque cordatis crenato-dentatis utrinque glabris vel ert parce Яоесово- tomentulosis et mox glabratis subinde purpurascenti- bus; foliis inferioribus plus minusve pa А et amplexieaulibus; inflo- rescentiis terminalibus corymboso-cymosis; capitulis paucis vel nu merosis 8-10 m. altis radiatis; involucro campanulato- cen glabro; bracteis aa 13-21 putes lanceolatis 5-6 mm. lo ongis acutis pend floseulis liguliferis 8-10, ligulis flavis; floribus disei numerosis; achae valde eostatis glabris.— Collected at Quebrada Honda, State of Durango, Martes 20 and 21 May, 1906, Palmer 213 (Gray Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.), TYPE. [Vor. 3 118 ANNALS OF THE MISSOURI BOTANICAL GARDEN An herbaceous perennial, glabrous or white-floccose in the leaf-axils and on the margins of the petioles; stem simple or branched, 1.5 to 4 dm. high from a stout rootstock, striate, glabrous; lower leaves petiolate, round-ovate to ovate-oblong, 1 to 6 em. long, 1 to 5 em. broad, usually cordate, crenate-den- tate, glabrous on both surfaces or slightly floecose-tomentulose in the early stages but soon glabrate; upper stem-leaves more or less pinnatisect and amplexicaul; inflorescence a subeorym- bose eyme terminating the stem and branches, few to many- headed; heads 8 to 10 mm. high, radiate; involucre campanu- late, ealyeulate, glabrous; bracts of the involucre 13 to 21, linear-lanceolate, attenuate, 5 to 6 mm. long; ray-flowers 8 to 10, rays pale yellow; disk-flowers rather numerous; achenes strongly ribbed, glabrous. Distribution: western Mexico. Specimens examined : Durango: Quebrada Honda, 20-21 May, 1906, Palmer 213 (Gray Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.), TYPE. 52. S. Pammelii Greenm.! An herbaceous perennial, glabrous or slightly white-tomen- tulose in the axils of the leaves; stems erect, 2 to 4 dm. high, and, as well as the petioles, more or less purplish towards the base; lower leaves long-petiolate, rotund-ovate, 1 to 2.5 em. long, nearly or quite as broad, shallowly cordate or subtrun- cate, crenate to nearly entire; upper stem-leaves petiolate and somewhat lyrate to sessile and pinnatisect to entire; inflo- ‘Senecio Pammelii Greenm. sp. nov., herbaceus p ubique glabrus vel in axillis foliorum paululo albo- e nir old eaulibus ereetis 2-4 dm. altis ad basin petiolis etiam y тое purpuraseentibus; 49 liis ылы longi-petiolatis rotu Fide atis em. longis et latis brevi-cordatis vel subtruneatis erenatis vel fer ét? ? toli 8 pit sala бе petiolatis et ae tis н. — us et Wee inforescentiis corymboso-eymosis; capitulis 7-10 mm. altis paucis vel pluribus radiatis; involuero Seite caly vo Wei ges icone ri 13-21 lineari-lanceola tis 5 6 mm. longis acutis glabris; flosculis A ris 10-12, ligulis avis; floribus disei numerosis; onda glabris.—Colleeted in Peterson Cañon, Peterson, Utah, alt. 2895 m., 19 July, 1902, Pammel d Blaokiocod 3870 (Мо, Bot. Gard. Herb.), TYPE; on Ruby Mou GE near Blaine post-office, Elko Co., alt. 2710 m., 27 Aug., 1913, Heller 11096 (U. S. Nat. Herb. and Mo. Bot. Z š зе Б species may be looked for in herbaria under S. aureus, also under 8. "in having been distributed under both names 1916] GREENMAN—MONOGRAPH OF SENECIO 119 rescence a few to many-headed corymbose суше; involucre campanulate, sparingly calyculate; heads 7 to 10 mm. high, radiate; braets of the involucre 13 to 21, linear-lanceolate, 5 to 6 mm. long, acute, glabrous; ray-flowers 10 to 12, rays bright yellow ; disk-flowers numerous ; achenes glabrous. Distribution: mountains of Utah and Nevada. Specimens examined: Utah: moist rocks and shady woods, Peterson Cañon, Peterson, alt. 2895 m., 19 July, 1902, Pammel £ Blackwood 3870 (Mo. Bot. Gard. Herb.), TYPE. Nevada: Ruby Mountains, near Blaine post-office, Elko Co., alt. 2710 m., 27 Aug., 1913, Heller 11096 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.). 53. S. aureus L. Sp. Pl. 2:870. 1753, ed. 2, 1220. 1763, and ed. 3, 1220. 1764; Michx. ЕІ. 2: 120. 1803; Willd. Sp. РІ. 3:1998. 1804; Pursh, Fl. Am. Sept. 2:530. 1814, and ed. 2, 1816; Nutt. Gen. 2:165. 1818; Ell. Sketch 2:331. 1824; Bigel. Fl. Bost. 307. 1824; Sprengl, Syst. Veg. 3:560. 1826, excl. syn.; DC. Prodr. 6:432. 1837; Torr. & Gray, Fl. N. Am. 2:442. 1843, in part; Torr. in Nicollet’s Report, App. B, 153 [237]. 1845; Gray, Syn. Fl. N. Am. 1? :391. 1884, and ed. 2, 1886, in part; Macoun, Cat. Canadian РІ. 264. 1884, in part; Millsp. Med. Pl. 1:91, pl. 91. 1892; Goodale, Wild Fls. of Am. 77, pl. 15. 1894; Britton & Brown, Ill. Fl. 3:480, fig. 4047. 1898, and ed. 2, 544, fig. 4625. 1913, in part; Greenm. in Rhodora 3:4. 1901; Monogr. Senecio, I. Teil, 23. 1901, and in Engl. Bot. Jahrb. 32:19. 1902; Britton, Manual 1028. 1901, and ed. 2, 1905; Greenm. in Gray, Manual, ed. 7, 854. 1908; Small & Carter, Fl. Lancaster County 310. 1913. S. tussilaginoides Walt. Fl. Carol. 208. 1788. S. rotundifolius Stokes, Bot. Mat. Med. 4:215. 1812. S. fastigiatus Schwein. ex Ell. Sketeh 2:331. 1824. An herbaceous perennial; stems one to several from a rela- tively slender rootstock, 3 to 6 dm. high, glabrous or not infre- quently white-tomentulose in the leaf-axils, along the margins of the petioles, and in the inflorescence; lower leaves petiolate, undivided and rotund-ovate, somewhat triangular-ovate to [Vor. 3 120 ANNALS OF THE MISSOURI BOTANICAL GARDEN oblong-ovate, 1 to 14 em. long, two-thirds to nearly or quite as broad, erenate to doubly serrate-dentate, usually deeply cordate at the base, green on both surfaces or tinged with purple beneath, glabrous or occasionally slightly tomentulose in the early stages and soon glabrate; petioles 1.5 to 25 em. long; stem-leaves variable, petiolate to sessile and amplexicaul, lyrate to pinnatisect, reduced towards the inflo- rescence sometimes to linear entire bracts; inflorescence a ter- minal several to many-headed corymbose cyme; heads 8 to 10 mm. high, radiate; involucre campanulate, calyculate, gla- brous or oceasionally slightly tomentulose; bracts of the invo- lucre (13-) 21, linear, acute, 6 to 8 mm. long; ray-flowers 8 to 12, rays yellow ; disk-flowers numerous ; achenes glabrous. Distribution: Labrador to Georgia, and west to North Dakota and Arkansas. Specimens examined. Labrador: shore of Seal Lake, 3 Aug., 1896, Spreadborough 14387 (Geol. Surv. Canada Herb.). Newfoundland: north of Placentia Junction, 11 Aug., 1894, Robinson € Schrenk (Gray Herb. and U. S. Nat. Herb.) ; banks of Salmonier River, 21 Aug., 1894, Robinson & Schrenk (Gray Herb, Geol. Surv. Canada Herb., апа U. S. Nat. Herb.) ; Benoist's Cove, Bay of Islands, July, 1895, Waghorne (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; between Port-aux-Basques and Bay of Islands, July, 1902, collector not indicated (Gray Herb.). Prince Edward Island: wet meadows, Tignish, 27 July, 1888, Macoun 14812 (Geol. Surv. Canada Herb.). Nova Scotia: Arcadia, Yarmouth Co., 29 June, 1911, Meth- eny (Phil. Acad. Nat. Sci. Herb.). New Brunswick: swamps, Tobique, July, 1884, Hay 14817 (Geol. Surv. Canada Herb.) ; along the St. John River, above Woodstock, 3 July, 1899, Macoun 22587 (Geol. Surv. Canada Herb.). Quebec: damp thickets, Lake Mistassini, 24 July, 1885, Macoun 14819 (Geol. Surv. Canada Herb.); Fort Chimo, on the Ungava River, 31 Aug., 1896, Spreadborough 14386 1916] GREENMAN—MONOGRAPH OF SENECIO 121 (Geol. Surv. Canada Herb.) ; in swamps and marshes, Anti- сова Island, 20 Aug. 1883, Macoun 14816 (Geol. Surv. Canada Herb.); swamps and sandy woods, River Ste. Anne des Monts, Gaspé, 19 Aug., 1882, Macoun 14821 (Geol. Surv. Canada Herb.) ; banks of Grand River, Gaspé Co., 30 June-3 July, 1904, Fernald (Gray Herb.) ; ealeareous alpine meadow, Table-top Mountain, Gaspé Co., alt. 1000-1125 m., 7 Aug., 1906, Fernald & Collins 260 (U. S. Nat. Herb.) ; coniferous forest, Low’s Trail from the Forks of River Ste. Anne des Monts to Table-top Mountain, Gaspé Co., alt. 550 m., Fernald & Collins 765, 766, and 767 (Gray Herb.); mossy arbor-vitae woods, east of Grande Coupe, Percé, Gaspé Co., 6 Aug., 1907, Fernald £ Collins 1208 (Gray Herb.) ; alluvial woods, mouth of Bon- aventure River, Bonaventure Со., 31 July, 1902, Williams Ф Fernald (Gray Herb.) ; wet alluvial shores, gravelly beaches and flats, between Baldé and the Baie des Chaleurs, Bonaven- ture River, 5-8 Aug., 1904, Collins, Fernald & Pease (Gray Herb.) ; alluvial thickets, between the Forks and Brile Brook, Little Caseapedia River, 29 and 30 July, 1904, Collins, Fer- nald Ф Pease (Gray Herb.). Ontario: in a bog, Rainy Lake, Algonquin Park, 12 June, 1900, Macoun 21869 (Geol. Surv. Canada Herb.) ; river banks, Inglewood, 4 July, 1890, White 14823 (Geol. Surv. Canada Herb.); swamps and sandy woods, Kammistiquia River, 13 July, 1869, Macoun 14814 (Geol. Surv. Canada Herb.) ; Glen Elgin, Lincoln Co., 10 June, 1897, McCalla 662, 22779 (Geol. Surv. Canada Herb.); swamps above Leamington, 30 May, 1901, Macoun 26671 (Geol. Surv. Canada Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; London, June, 1884, Burg- ess (U. Б, Nat. Herb.) ; Amherstburg, 9 June, 1882, Macoun 14813 (Geol. Surv. Canada Herb.). Maine: wet thickets, Limestone, Aroostook Co., 20 June, 1898, Fernald 2403 (Gray Herb.) ; arbor-vitae swamp, Presque Isle, 12 July, 1902, Williams, Collins € Fernald (Gray Herb.) ; cedar swamp and clearings, Blaine, 23 June, 1898, Fernald 2404 (Gray Herb.). [Vor. 3 122 ANNALS OF THE MISSOURI BOTANICAL GARDEN New Hampshire: Greenville, 6 June, 1897, Fernald (Gray Herb.); sphagnum bog, Jaffrey, 10 July, 1897, Robinson 590 (Gray Herb.). Vermont: Peacham, 1 July, 1888, F. Blanchard (Mo. Bot. Gard. Herb.) ; Brandon, 8 June, 1883, Knowlton (U. S. Nat. Herb.); Castleton, 3 June, 1898, Eggleston (Mo. Bot. Gard. Herb.); Windham, 25 July, 1901, W. H. Blanchard (Gray Herb.) ; Manchester, 27 June, 1898, Day 102 (Gray Herb. and U. S. Nat. Herb.). Massachusetts: Hamilton, Essex Co., Oakes (Gray Herb.) ; near Boston, 13 June, 1854, ex Herb. Wm. Boot (Gray Herb.) ; without definite locality, Pickering (Phil Acad. Nat. Sci. Herb.) ; Boston, ex Herb. Short (Phil. Acad. Nat. Sci. Herb.) ; Arlington Heights, 5 June, 1904, Greenman 3005 (Mo. Bot. Gard. Herb.); Coneord, 30 May, 1896, Greenman 305 (Mo. Bot. Gard. Herb.) ; Purgatory Swamp, Dedham, 30 May, 1897, Greenman 280 (Mo. Bot. Gard. Herb.); Holbrook, 18 June, 1899, Greenman 609 (Mo. Bot. Gard. Herb.) ; South Framing- ham, 18 May, 1890, Sturtevant (Mo. Bot. Gard. Herb.) ; Non- quitt, 21-30 May, 1889, Sturtevant (Mo. Bot. Gard. Herb.) ; Southampton, ex Herb. Chapman (Mo. Bot. Gard. Herb.). Rhode Island: Monns Swamp, Providence, 24 May, 1891, Collins (Gray Herb.); Providence, coll. of 1846, Thurber (Gray Herb.), 10 June, 1900, Chamberlain 154 (U. 5. Nat. Herb.), and without date, Olney (Kew Herb.). Connecticut: Southington, 30 May, 1896, Andrews 4 (Gray Herb.), 25 May, 1897, Bissell 112, 1480 (Mo. Bot. Gard. Herb.) ; Waterbury, З June, 1888, Du Bois (U. S. Nat. Herb.) ; New Haven, without date, Eaton (Gray Herb.) ; near Maltby Park, New Haven, 28 May, 1884, Safford 85 (U. S. Nat. Herb.) ; Bridgeport, 25 May, 1896, Eames (U. S. Nat. Herb.). New York: Lebanon Springs, 31 May, 1898, Harrison 8 (U. S. Nat. Herb.) ; vieinity of North Harpersfield, Delaware Co., June, 1906, Topping 69, 103, 106 (0. 8. Nat. Herb.); vieinity of Oneida, June-July, 1914, Maxon (U. S. Nat. Herb.) ; near Syracuse, Straub (U. S. Nat. Herb.) ; Ithaca, 3 and 21 June, 1885, Coville, 21 June, 1889, Pearce, and 31 1916] GREENMAN—MONOGRAPH OF SENECIO 192 Мау, 1890, Rowlee (U. S. Nat. Herb.) ; near Ithaca, coll. of 1889, Norris (Mo. Bot. Gard. Herb.); Cascadilla Creek, 6 June, 1877, Trelease (Mo. Bot. Gard. Herb.) ; Fall Creek, 14 June, 1893, Schrenk (Mo. Bot. Gard. Herb.); western New York, collector and date not indicated (Gray Herb.); Clove Lake, Staten Island, 28 May, 1905, Dowell 3764 (Mo. Bot. Gard. Herb.). New Jersey: New Durham, Brownne (Kew Herb.) ; Tena- fly, 26 May, 1894, Pollard (С. S. Nat. Herb.) ; Fairview, 23 May, 1895, Van Sickle (U. S. Nat. Herb.) ; Charlotteburg, 24 May, 1908, Mackenzie 3085 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Newfoundland, 14 June, 1908, Mackenzie 3127 (U. S. Nat. Herb.) ; Mt. Tabor, 16 June, 1907, Mackenzie 2640 (Mo. Bot. Gard. Herb.) ; Budd's Lake, 28 May, 1895, Heritage (Phil. Aead. Nat. Sci. Herb.); Bound Brook, 31 May, 1904, House (U. S. Nat. Herb.); river bank above Crosswick's Creek, 29 May, 1904, Williamson (C. S. Williamson Herb.) ; Batsto, 22 Мау, 1912, Bassett 17 (Phil Acad. Nat. Sci. Herb.) ; shore of Delaware River, Camden Co., 10 May, 1910, Long 3259 (Phil. Acad. Nat. Sci. Herb.); Washington Park, 9 May, 1897, Jahn (Phil. Acad. Nat. Sci. Herb.) ; below Wash- ington Park, 15 May, 1895, Jahn (Phil. Acad. Nat. Sei. Herb.) ; Glassboro, 14 May, 1910, Long 3346 (Phil. Acad. Nat. Sci. Herb.); Bennett, 29 Oct., 1912, Long 7954 (Phil. Acad. Nat. Sei. Herb.) ; without locality or date, Read (Phil. Acad. Nat. Sci. Herb.). Pennsylvania: Milford, May, 1905, Mell (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Tobyhanna Mills, 3 July, 1893, Crawford (Phil. Acad. Nat. Sci. Herb.) ; in deep shaded bog, Tobyhanna, 27 July, 1907, Bartram (Phil. Acad. Nat. Sei. Herb.); bog near Easton, 18 May, 1894, Tyler (Phil. Acad. Nat. Sci. Herb.) ; Hellertown, ex Herb. Detwillert (Phil. Acad. Nat. Sei. Herb.); streamlet east of Slatington reservoir, 2 June, 1912, Pretz 4512 (Phil. Acad. Nat. Sci. Herb.) ; Three- mile Run, 21 May, 1886, Fretz (Phil. Acad. Nat. Sci. Herb.) ; Rock Hill, 2 June, 1889, Pollard (U. S. Nat. Herb.) ; Sellers- ville, June, 1887, F'retz (Phil. Acad. Nat. Sci. Herb.) ; Doyles- [Vor. 3 124 ANNALS OF THE MISSOURI BOTANICAL GARDEN town, 18 May, 1883, Pond (С. 5. Nat. Herb.) ; Woodbourne, 30 May, 1904, Brown (Phil. Acad. Nat. Sci. Herb.) ; Tullytown, 30 Мау, 1899, Williamson (C. S. Williamson Herb.) ; Tully- town, 3 May, 1899, Crawford (Phil. Acad. Nat. Sci. Herb.) ; Tullytown, 20 May, 1899, Fretz (Phil. Acad. Nat. Sci. Herb.) ; Bucks Co., coll. of 1867, Allen (Phil. Acad. Nat. Sci. Herb.) ; Shannonville, 12 June, 1891, Crawford (Phil. Acad. Nat. Sci. Herb.) ; Arcola, 6 May, 1892, Crawford (Phil. Acad. Nat. Sci. Herb.); Merion, 25 April, 1871, Redfield (Mo. Bot. Gard. Herb.) ; French Creek Falls, MacElwee (Phil. Acad. Nat. Sci. Herb.) ; Darby Creek, near Paoli, 21 May, 1905, Bartram (Phil. Acad. Nat. Sei. Herb.) ; Westtown Meadow, 7 May, 1890, N. H. C. (Phil. Acad. Nat. Sei. Herb.); near Pocopson, 27 May, 1904, Painter 628 (Mo. Bot. Gard. Herb.) ; near Brandy- wine, Townsend (Phil. Acad. Nat. Sci. Herb.) ; New Garden, 29 May, 1904, Vanatta (Phil. Acad. Nat. Sci. Herb.) ; Haver- ford, D. B. Smith (Kew Herb.) ; west side of Wissahickon River, 18 June, 1834, collector not indicated (Phil. Acad. Nat. Sei. Herb.); West Philadelphia, 4 May, 1890, Mac- Elwee 475 (Phil. Acad. Nat. Sci. Herb.); Crum Creek, near Philadelphia, 6 June, 1867, Redfield (Mo. Bot. Gard. Herb.) ; Crum Creek, 30 May, 1898, Githens (Phil. Acad. Nat. Sci. Herb.) ; Byberry, coll. of 1862, Martindale (Phil. Acad. Nat. Sci. Herb.); near Mickelton, 14 May, 1892, Heritage (Phil. Acad. Nat. Sci. Herb.) ; Delaware Co., 10 May, 1891, MacElwee (Phil. Acad. Nat. Sei. Herb.); Clifton, 7 May, 1893, Lloyd (Phil. Acad. Nat. Sei. Herb.) ; Dillerville swamp, Lancaster Co., 30 May, 1901, Heller (Gray Herb., U. 8. Nat. Herb., and Mo. Bot. Gard. Herb.); on Little Conestoga near Stoneroad’s Mill, 23 May, 1901, Heller (Gray Herb. and U.S. Nat. Herb.); between York Furnace and Tuequam, 11 May, 1901, Heller (Gray Herb. and U. S. Nat. Herb.); Fishing Creek, 16 May, 1906, Carter (Phil. Acad. Nat. Sci. Herb.) ; west branch of Octoraro Creek, 6 May, 1891, Small & Heller (U. S. Nat. Herb.) ; York Furnace, York Co., 13 May, 1899, MacElwee 201 (Phil. Acad. Nat. Sei. Herb.) ; Meadow-ground Mountain, Fulton Co., 4 June, 1905, Stone 226 (Phil. Acad. 1916] GREENMAN—MONOGRAPH OF SENECIO 125 Nat. Sei. Herb.) ; Mt. Alto, coll. of 1900, Писк (Мо. Bot. Gard. Herb.). Delaware: Granogue, 27 May, 1896, Commons (Phil. Acad. Nat. Sei. Herb.); near Centerville, 26 May, 1875, Commons (Phil. Acad. Nat. Sei. Herb.) ; Green Bank, 12 May, 1884, Com- mons (Phil. Acad. Nat. Sci. Herb.) ; Elsmere, 14 May, 1894, Commons (Phil. Acad. Nat. Sci. Herb.); west branch of Naaman's Creek, 15 May, 1909, Pennell 2040 (Phil. Acad. Nat. Sei. Herb.). Maryland: along the Susquehanna River, Cecil Co., 18 April, 1913, St. John & Long 8076 (Phil. Acad. Nat. Soi. Herb.) ; Calreston, 5 May, 1889, Thurston (U.S. Nat. Herb.) ; near Great Falls, Montgomery Co., 18 May, 1900, M. F. Miller (U. S. Nat. Herb.) ; Plummer's Island, near Cabin John, 30 April, 1902, Kearney & Maxon (U.S. Nat. Herb.); Mont- gomery Co., near Washington, Batchelder (Phil. Acad. Nat. Sci. Herb.) ; Marlboro, 6 May, 1900, Morris 914 (U. S. Nat. Herb.) ; Savage River, Garrett Co., 25 April, 1897, Knowlton (U. S. Nat. Herb.). Distriet of Columbia: Washington, coll. of 1873, Vasey (U. S. Nat. Herb.) ; High Island, 2 April, 1876, Ward (U. S. Nat. Herb.); vieinity of Washington, 10 June, 1877, Ward (U. S. Nat. Herb.); without definite locality, 29 April, 1888, Burgess (U. S. Nat. Herb.); National Park, 6 May, 1892, F. Blanchard (Mo. Bot. Gard. Herb.) ; Glen Echo, 24 April, 1895, Pollard 89 (U. S. Nat. Herb.) ; Rock Creek Park, 4 May, 1896, Steele (Gray Herb. and U. S. Nat. Herb.) ; High Island, April, 1898, Williamson (C. S. Williamson Herb.); pine woods, Daleeartia Reservoir, 28 May, 1905, Painter 1305 (Mo. Bot. Gard. Herb.); Chevy Chase, 16 May, 1905, House 728 (Mo. Bot. Gard. Herb.). Virginia: Arlington, April, 1891, F. Blanchard (Mo. Bot. Gard. Herb.) ; Stony Man Mountain, 3 July, 1903, G. S. Miller (U. S. Nat. Herb.) ; on Bear Creek, east of Hungry Hollow, Smyth Co., alt. 880 m., 7 June, 1892, Small (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; about Chatham Hill Gap, Walker Mountain, alt. 915 m., Small (Gray Herb.) ; Walker Mountain, [VoL. 3 120 ANNALS OF THE MISSOURI BOTANICAL GARDEN alt. 640 m., E. G. Britton & A. M. Vail (Phil. Acad. Nat. Sci. Herb.). North Carolina: Cloudland, Roan Mountain, 25 June, 1902, Cannon 10 (U.S. Nat. Herb.) ; vicinity of Asheville, May, 1888, McCarthy (U.S. Nat. Herb.) ; Biltmore, 24 April, 1896, and 13 April, 1897, Biltmore Herb. 889, 889a (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; mountains of North Carolina, Ashe (U. S. Nat. Herb.). Georgia: without locality, ex Herb. Chapman 2340 (Mo. Bot. Gard. Herb.). West Virginia: Upshur Co., 4 May, 1895, and 27 May, 1897, Pollock (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.). Ohio: Berea, May, 1895, Ashcroft (Mo. Bot. Gard. Herb.) ; Canton, May, 1835, Riehl (Mo. Bot. Gard. Herb.) ; Columbus, Sullivant (Kew Herb.) ; Champaign Co., 27 May, 1893, Werner 188 (Gray Herb.). Michigan: Port Huron, 18 Мау, 1896, Dodge 296 (0. 8. Nat. Herb.); moist rich woods and river banks, Rochester, May, 1914, Farwell 3384, 3628V5, and at Parkdale Farms, colls. of May and June, 1912, 1913, Farwell 2552, 3408, 3425 (Mo. Bot. Gard. Herb.) ; Owosso, 21 May, 1889, Hicks (U. S. Nat. Herb.). Indiana: Chain Lakes, 6 May, 1913, Niewwland 11005 (Mo. Bot. Gard. Herb.) ; Mineral Springs, 20 May, 1912, Niewwland 10019 (Mo. Bot. Gard. Herb.) ; bank of Wabash River, west of Shively Bridge, 19 May, 1901, Mackenzie (U. S. Nat. Herb.) ; West Lafayette, 10 May, 1912, Overholts (Mo. Bot. Gard. Herb.) ; Jefferson Co., Hubbard (Gray Herb.) ; banks of Lick Creek near Abby Dell, 25 May, 1901, Mackenzie (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.). Kentucky: Blue-licks and Mud-lick, Short (Phil. Acad. Nat. Sci. Herb.) ; Bowling Green, May, 1892, Price (Mo. Bot. Gard. Herb.). Illinois: in moist rich thickets and wet meadows near Beach, 16 June, 1907, Greenman 2007, 2022, 2023 (Mo. Bot. Gard. Herb.) ; near Woodlawn, Jefferson Co., 16 Мау, 1898, Eggert (Mo. Bot. Gard. Herb.). Minnesota: Bear Creek, May, 1890, Holzinger (U. S. Nat. Herb.). 1916] GREENMAN—MONOGRAPH OF SENECIO IET South Dakota: Brookings, coll. of 1892, Williams (U.S. Nat. Herb.); along creeks, Brookings, 26 May, 1894, T'hornber (Mo. Bot. Gard. Herb.) ; low ground, Oakwood, 23 May, 1902, A. G. J. (Mo. Bot. Gard. Herb.) ; Custer Co., alt. 1675 m., 16 July, 1892, Rydberg 827 (Gray Herb.), in part. Nebraska: Emerson, 12 June, 1893, Clements 2513 (U. S. Nat. Herb.). Iowa: Iowa City, Hitchcock (Mo. Bot. Gard. Herb.); prairies, near Council Bluffs, Nicollet’s North-Western Ex- pedition, 16 May, 1839, Geyer 97 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.). Missouri: low ground, Adair Co., 5 May, 1884, Sheldon (Mo. Bot. Gard. Herb.) ; Courtney, coll. of 1880, Bush (U. 8. Nat. Herb.) ; banks of River des Peres, near St. Louis, Мау, 1883, Engelmann (Mo. Bot. Gard. Herb.) ; “Valley Forge", 11 May, 1888, Pammel (Mo. Bot. Gard. Herb.) ; Glencoe, 22 Мау, 1879, Eggert (Mo. Bot. Gard. Herb.) ; Allenton, April, 1890, Letterman (Mo. Bot. Gard. Herb.) ; Allenton, 23 Мау, 1892, Glatfelter 293 (Mo. Bot. Gard. Herb.) ; Cliff Cave, 26 Aug., 1898, Norton (Mo. Bot. Gard. Herb.) ; Pacific, 4 July, 1879, Eggert (Mo. Bot. Gard. Herb.) ; Pacifie, 15 May, 1900, Norton (Mo. Bot. Gard. Herb.) ; Victoria, 10 May, 1890, Hitch- cock (Mo. Bot. Gard. Herb.) ; near Sunnyside, 22 May, 1879, Eggert (Mo. Bot. Gard. Herb.) ; De Soto, 22 May, 1892, Eggert (Mo. Bot. Gard. Herb.) ; Valles Mines, May, 1835, (?) Engel- mann (Mo. Bot. Gard. Herb.) ; Shannon Co., coll. of 1890, Bush (0. S. Nat. Herb.) ; Monteer, 24 May, 1900, Bush 370 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; along streams, Monteer, 14 May, 1901, and 29 April, 1907, Bush 465, 4395 (Mo. Bot. Gard. Herb.) ; Greene Co., 30 May, 1879, Shepard (Gray Herb.) ; Swan, Taney Co., 21 April, 1907, Bush 4225 (Mo. Bot. Gard. Herb.) ; Galena, Stone Co., 3 May, 1914, E. J. Palmer 5743 (Mo. Bot. Gard. Herb.); Reding's Mill, 9 April, 1909, E. J. Palmer 1655 (Mo. Bot. Gard. Herb.). Arkansas: Eureka Springs, Carroll Co., 15 May, 1914, E. J. Palmer 5623 (Mo. Bot. Gard. Herb.) ; northwestern Arkan- sas, April, 1880, Harvey 45 (Gray Herb.). [Vor. 3 128 ANNALS OF THE MISSOURI BOTANICAL GARDEN Var. gracilis (Pursh) Britt. in Britton & Brown, Ill. КІ. 3:481. 1898; Britton, Manual 1028. 1901, and ed. 2, 1905; Greenm. Monogr. Senecio, I. Teil, 23. 1901, in Emgl. Dot. Jahrb. 32:19. 1902, and in Gray, Manual, ed. 7, 854. 1908; Porter, Fl. Penn. 339. 1903. S. gracilis Pursh, Fl. Am. Sept. 2:529. 1814, and ed. 2, 1816; DC. Prodr. 6:432. 1837; Small, Fl. Southeastern U. S. 1303. 1903, and ed. 2, 1913. Stems slender; basal leaves relatively small, those of the offshoots, as well as the lower stem-leaves, rotund-ovate to oblong-ovate, 1 to 2 em. long and nearly to quite as broad. In all other essential eharaeters like the species into whieh it direetly passes. Distribution: occurring with the species, but especially in moist open places. Specimens examined : Ontario: Squirrel Island, 10 June, 1904, Dodge 14, 2974 (U. S. Nat. Herb.). New Jersey: Point Pleasant, Williamson (C. S. William- son Herb.) ; meadows, ete., New Egypt, 14 May, 1906, Grove 315 (Phil. Acad. Nat. Sci. Herb.) ; Spray, de Chalmot (U. 8. Nat. Herb.); Swedesboro, 15 Мау, 1892, Lippincott (Phil. Acad. Nat. Sei, Herb.). Pennsylvania: Wayne, 2 May, 1908, Bartram (Phil. Acad. Nat. Sei. Herb.); Lehigh Co., Pretz 345, 4398, 3272, 6483, 6457 (Phil. Acad. Nat. Sei. Herb.); Bucks Co., coll. of 1860, Krout (Phil. Acad. Nat. Sci. Herb.); Nockamixon, coll. of 1893, Crawford (Phil. Acad. Nat. Sei. Herb.) ; near Quaker- town, 9 May, 1899, Fretz (Phil. Aead. Nat. Sci. Herb.) ; Rock Hill, 31 May, 1903, MacElwee (Phil. Acad. Nat. Sci. Herb.) ; Tohickon Creek near Doylestown, 30 May, 1902, Brown (Phil. Acad. Nat. Sci. Herb.); meadows along Wissahickon Creek, Fort Washington, 7 May, 1909, Long (Phil. Acad. Nat. Sci. Herb.) ; near Ardmore, 15 Мау, 1909, Eckfeldt (Phil. Acad. Nat. Sci. Herb.); near Tredyffrin, 6 May, 1906, Bartram (Phil. Acad. Nat. Sci. Herb.) ; Chester Heights, 9 May, 1909, Pennell 2030 (Phil. Acad. Nat. Sci. Herb.) ; Dillerville Swamp, 1916] GREENMAN—MONOGRAPH OF SENECIO 129 Lancaster Co., 25 May, 1889, Heller (Gray Herb.); New Providence, 18 May, 1900, Heller (Mo. Bot. Gard. Herb.) ; about Penryn, Lebanon Co., 27 May, 1893, Heller & Halbeck 876 (Gray Herb., U.S. Nat. Herb., and Mo. Bot. Gard. Herb.). Delaware: west branch of Naaman’s Creek, 15 May, 1909, Pennell 2044 (Phil. Acad. Nat. Sci. Herb.) ; near Cooch’s Mill, 29 May, 1896, Commons (Phil. Acad. Nat. Sci. Herb.). Maryland: near Great Falls, 8 May, 1895, Mearns (С. 8. Nat. Herb.). Distriet of Columbia: Carberry Meadows, 7 May, 1903, Steele (Mo. Bot. Gard. Herb.). Michigan: marl beds on Parkdale Farm, 25 May, 1913, Far- well 3415 (Mo. Bot. Gard. Herb.); in wet meadows, Grand Rapids, 17 May, 1894, and 26 June, 1897, Cole (Gray Herb.). Illinois: Palos Park, 22 May, 1913, Millspaugh 3744 (Field Mus. Herb.). Minnesota: Fort Snelling, coll. of 1888, Forwood, also 1 and 14 June, 1891, Mearns (U. S. Nat. Herb.) ; Nicollet, June, 1892, Ballard (U. S. Nat. Herb.). North Dakota: in bogs, Butte, 3 June, 1906, Lunell (U. S. Nat. Herb.). Var. semicordatus (Mack. & Bush) Greenm. comb. nov. S. semicordatus Mack. & Bush, Mo. Bot. Gard. Ann. Rept. 16:107. 1905. S. aureus Mack. & Bush, Manual Fl. Jackson County, Mo. 207. 1902, in part, not L. S. aureus? > Balsamitae Greenm. Rhodora 10:69, 1908. Lower leaves rotund-ovate to oblong-ovate, 1 to 8 em. long, 1 to 4 em. broad, usually rounded at the apex, shallowly eor- date. Distribution: beaches, shores, and prairies. Eastern Quebec, Illinois, and Missouri. Specimens examined: Quebee: wet alluvial shores, gravelly beaches and flats, be- tween Baldé and the Baie des Chaleurs, Bonaventure River, 5, 6, and 8 Aug., 1904, Collins, Fernald & Pease (Gray Herb., photograph in Field Mus. Herb. and Mo. Bot. Gard. Herb.). 9 [Vor. 3 130 ANNALS OF THE MISSOURI BOTANICAL GARDEN Illinois: in wet meadows near Beach, Lake Co., 16 June, 1907, Greenman 2022 (Field Mus. Herb., photograph in Mo. Bot. Gard Herb.), in part; low ground, Evanston, Earl, also Price (U. S. Nat. Herb.). Missouri: on prairies, Levasy, Jackson Co., 18 May, 1902, Bush 1678 (Gray Herb. and Mo. Bot. Gard. Herb.), түрЕ; swales on prairies, Levasy, 11 May, 1904, Bush 1940 (Mo. Bot. Gard. Herb.). Although some of the specimens cited show indieations of a possible origin by hybridization, as was indicated in ‘Rhodora’ in 1908, yet the examination of additional material points rather towards an origin by variation. 54. 8. Robbinsii Oakes ex Rusby, in Bull. Torr. Bot. Club 20:19, pl. 139. 1893; Heller, Cat. N. Am. РІ, 147. 1898, and ed. 2, 230. 1900; Britton & Brown, Ill. КІ. 3:480, fig. 4046. 1898, and ed. 2, 544, fig. 4624. 1913; Greenm. in Rhodora 3:4. 1901; Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Small, Fl. Southeastern U. S. 1303. 1903, and ed. 2, 1913; Kennedy in Rhodora 6:133. 1904; Britton, Manual 1028. 1901, and ed. 2, 1905; Greenm. in Gray, Manual, ed. 7, 854. 1908. S. aureus var. lanceolatus Oakes ex Torr. & Gray, Fl. N. Am. 2:442. 1843; Macoun, Cat. Canadian Pl. 265. 1884. S. aureus var. Balsamitae Gray, Syn. Fl. N. Am. 1:391. 1884, and ed. 2, 1886, in part, as to S. aureus var. lanceolatus Oakes in synonymy. An herbaceous perennial, glabrous or slightly tomentose along the margins of the petioles of the leaves, especially near their sheathing base; lower leaves long-petiolate, ovate-rotund to oblong-lanceolate, 1 to 10 em. long, 1 to 3 em. broad, erenate to sharply and more or less serrate-dentate, cordate to ab- ruptly narrowed at the base, green and glabrous on both sur- faces; upper stem-leaves petiolate and sublyrate or sessile and more or less ріппайвесі; inflorescence a terminal several to many-headed corymbose есуше; heads 8 to 10 mm. high, radi- ate; involucre campanulate, ealyeulate; bracts of the in- voluere about 21, linear, acute, glabrous; ray-flowers 10 to 12, rays yellow; disk-flowers numerous; achenes glabrous. 1916] GREENMAN—MONOGRAPH OF SENECIO 131 Distribution: Nova Scotia and Quebec, south to northern New England and New York; also on Roan Mt., North Caro: lina. Specimens examined : Nova Seotia: Margaree, Big Intervale, Cape Breton Island, 19 July, 1898, Macoun 19717 (Geol. Surv. Canada Herb.); Baddeck, Cape Breton Island, 10 July, 1898, Macoun 19718 (Geol. Surv. Canada Herb.); Boylston, July, 1890, Hamilton 22333 (Geol. Surv. Canada Herb.); in swamps and ditches, Truemanville, 30 July, 1883, T'rueman 14773 (Geol. Surv. Can- ada Herb.); railroad diteh, Truro, 27 July, 1911, Bartram (Phil. Aead. Nat. Sci. Herb.). Prince Edward Island: rocky places, Winslow Road, 16 July, 1888, Macoun 14794 (Geol. Surv. Canada Herb.). пеһес: swamp near Georgeville, Lake Memphremagog, 12 July, 1902, Churchill (Gray Herb.). Maine: dry thicket, Van Buren, 24 July, 1893, Fernald (Gray Herb.) ; open swampy woods, Aroostook Co., Mackenzie 3633 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); damp thickets, Cutler, 5 June, 1902, Kennedy, Williams, Collins & Fernald (Gray Herb.) ; wet meadow, Fitzgerald Pond, near Moosehead Lake, 6 July, 1895, Fernald 272 (Gray Herb., Г. 8. Nat. Herb., Greene Herb., and Mo. Bot. Gard. Herb.) ; meadow near High Head, Mt. Desert, 1 July, 1891, Redfield (Phil. Acad. Nat. Sci. Herb. and Mo. Bot. Gard. Herb.) ; Winthrop, coll. of 1862, Sturtevant (Mo. Bot. Gard. Herb.) ; Hartford, June, 1885, and July, 1892, Parlin (Gray Herb.) ; Westbrook, July, 1900, Ricker 672 (U. S. Nat. Herb.). New Hampshire: new gate of Notch, White Mountains, 7 July, 1878, E. € C. E. Faxon (Gray Herb.) ; Crawford Notch, 1 July, 1898, Greemnan 1105 (Gray Herb. and Mo. Bot. Gard. Herb.); wet meadow, Glen House, 10 July, 1910, Williamson 1421 (C. S. Williamson Herb.); Jackson, 26 July, 1890, Churchill (U. S. Nat. Herb.) ; Jackson, 10 Sept., 1896, Purdie (Gray Herb.) ; Llandoff Valley Meadows, Franconia, 18 June, 1895, E. Ф C. E. Faxon (Gray Herb. and U. 5. Nat. Herb.) ; cedar swamp, Franconia, 19 June, 1895, E. & C. E. Faxon (Gray Herb.). [Vor. 3 132 ANNALS OF THE MISSOURI BOTANICAL GARDEN Vermont: cedar swamp, Willoughby Lake, 26 July, 1885, Deane (Gray Herb.); Willoughby Mountain, 15 July, 1906, Williamson (C. S. Williamson Herb.); Stowe, July, 1899, Churchill € Greenman 294 (Mo. Bot. Gard. Herb.) ; Peacham, July, 1892, F. Blanchard (Mo. Bot. Gard. Herb.) ; Starksboro, 10 June, 1898, Eggleston (Mo. Bot. Gard. Herb.); Middle- bury, 23 and 25 June, 1883, Brainerd (Gray Herb) ; Pittsford, 14 June, 1902, Eggleston 2783 (Phil. Acad. Nat. Sci. Herb.) ; on cold bog, Rutland, 21 June, 1899, Eggleston 1383 (U.S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Mendon, 2 July, 1900, Eggleston 2040 (Mo. Bot. Gard. Herb.); Windham, coll. of 1901, W. H. Blanchard (Gray Herb.). New York: Stony Creek Ponds, Adirondack Mountains, 29 June, 1899, Rowlee, Wiegand & Hastings (Gray Herb.) ; north woods, Herkimer Co., coll. of 1864, Paine (Gray Herb.) ; meadow, South Branch, Herkimer Co., Aug., 1879, Tweedy (U.S. Nat. Herb.), form with somewhat less elongated leaves. North Carolina: summit of Roan Mountain, Mitchell Co., ай. 1920 m., Small € Heller 234 (Mo. Bot. Gard. Herb.). 55. S. pseudaureus Rydb. Bull. Torr. Bot. Club 24:298. 1897 ; ibid. 27: 180, pl. 5, fig. 10. 1900; Mem. N. Y. Bot. Gard. 1:446. 1900; ЕІ. Colo. 397. 1906; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Piper, Contr. U. S. Nat. Herb. 11:598. 1906; Nels. in Coulter & Nel- son, Manual Cent. Rocky Mountains 582. 1909, in part; Daniels, Univ. Mo. Studies, Sci. Ser. 2:252. 1911; Wooton & Standley, Contr. U. S. Nat. Herb. 19:747. 1915. An herbaceous perennial; stems erect, 3 to 7 dm. high, white- tomentulose in the axils of the leaves, along the margins of the petioles towards their base, and in the inflorescence, other- wise glabrous or nearly so; lower leaves long-petiolate, ovate- rotund to oblong-ovate, 1 to 10 em. long, 1 to 6 em. broad, rounded to acute at the apex, shallowly cordate, erenate to doubly serrate with somewhat ineurved teeth; petioles 1 to 23 em. long; stem-leaves petiolate or sessile, undivided or sublyrate to pinnatisect, and usually with rather sharply and doubly serrate-dentate and frequently revolute margins, more 1916] GREENMAN—MONOGRAPH OF SENECIO 133 or less attenuate; inflorescence a terminal few to many-headed corymbose eyme; heads 8 to 10 mm. high, radiate; involucre eampanulate, ealyeulate; bracts of the involucre (13-) 21, linear, acute, 6 to 8 mm. long, glabrous except at the penicillate tip; ray-flowers 10 to 13, rays pale yellow; disk-flowers numer- ous; achenes glabrous. Distribution: Saskatehewan, Alberta and British Columbia, south to New Mexico and California. Specimens examined: Saskatchewan: District of Assiniboia, 1 Aug., 1901, Wil- liamson (C. S. Williamson Herb.). Alberta: Laggan, Rocky Mountain Park, 13 July, 1904, Macoun 65016 (Gray Herb. and Geol. Surv. Canada Herb.); trail to Burgess Pass, Yoho Valley, 28 Aug., 1904, Macoun 65017 (Gray Herb. and Geol. Surv. Canada Herb.); Lake Louise, alt. 1830-2135 m., 17 July, 1906, Brown 559 (Phil. Acad. Nat. Sci. Herb.); near Banff, 6 July, 1891, Macoun (U. S. Nat. Herb. 232000); Banff, alt. 1340 m., 7 July, 1907, Butters Ё Holway 66 (U.S. Nat. Herb.) ; in swampy places, summit of South Kootenai Pass, 9 Aug., 1881, Dawson 14768 (Geol. Surv. Canada Herb.); in wet places, North Kootenai Pass, 28 July, 1883, Dawson 14815 (Geol. Surv. Canada Herb.). British Columbia: Burgess Trail, vicinity of Field, alt. 1545-1830 m., 29 June and 16 July, 1906, Brown 518 (Phil. Acad. Nat. Sci. Herb.) ; in woods, Emerald Lake, alt. 1310 m., 4 Aug., 1904, Petersen 148 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.) ; Bonaparte River, 18 June, 1889, Macoun (U. S. Nat. Herb. 219791), in part; Mount St. Thomas, between Kettle and Co- lumbia Rivers, 8 Aug., 1902, Macoun 64994 (Gray Herb. and Geol. Surv. Canada Herb.); grassy thickets, Guichon Creek, 8 July, 1888, Dawson 14769 (Geol. Surv. Canada Herb.) ; damp places, between North Thompson and Bonaparte Rivers, alt. 1220 m., 18 June, 1889, Macoun 14770 (Geol. Surv. Canada Herb.), in part; Trail, 9 June, 1902, Macoun 64993 (Geol. Surv. Canada Herb.). [Vor. 3 134 ANNALS OF THE MISSOURI BOTANICAL GARDEN Montana: Midvale, 1 July, 1903, Umbach 238 (U. S. Nat. Herb.) ; Big Fork, Flathead Co., 14 June, 1904, Jones (U.S. Nat. Herb.) ; Big Fork, 15 July, 1908, and Swan Lake, 25 Aug., 1908, Clemens (Field Mus. Herb. 376698 and 384958); Mac- Dougal Peak, 31 July, 1908, Clemens (Mo. Bot. Gard. Herb.) ; Little Belt Pass, 10 Aug., 1896, Flodman 918 (Mo. Bot. Gard. Herb.), со-түрє; Spanish Basin, Gallatin Co., alt. 1980 m., 28 June, 1897, Rydberg & Bessey 5263 (Gray Herb., Berlin Herb., and N. Y. Bot. Gard. Herb.) ; Gallatin Basin, alt. 2130 m., 5 Aug., 1905, Blankenship 291 (U. S. Nat. Herb., Mo. Bot. Gard. Herb., and Phil. Aead. Nat. Sei. Herb.). Yellowstone National Park: Lone Star Geyser Basin, 7 Aug., 1897, Rydberg & Bessey 5262 (U. S. Nat. Herb. and N. Y. Bot. Gard. Herb.) ; Gibbon Cañon, 28 Aug., 1899, A. £ E. Nelson 6748 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Yellowstone Falls, 5 Aug., 1885, Letterman 77 (Mo. Bot. Gard. Herb.); without definite locality, 5 Aug., 1902, Mearns 2860, 2978 (U. S. Nat. Herb.); Amethyst Creek, 14 Aug., 1887, Knowlton (U. S. Nat. Herb. 201412). Colorado: open wet meadow, below Estes Park, 5 July, 1912, Churchill (J. R. Churchill Herb.) ; Wagon Wheel Gap, Mineral Co., July, 1882, B. H. Smith (Phil. Acad. Nat. Sci. Herb.) ; Mancos, 21 June, 1898, Baker, Earle & Tracy 45 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.). New Mexico: vicinity of Chama, Rio Arriba Co., alt. 715— 870 m., 9 July, 1911, Standley 6635 (U. S. Nat. Herb.) ; along the Pecos River, east of Glorieta, San Miguel Co., alt. 1980 m., 4. & E. Heller 3682 (U.S. Nat. Herb. and Mo. Bot. Gard. Herb.). Idaho: West Fork of Priest River, alt. 900 m., 4 Aug., 1897, Leiberg 2825 (U.S. Nat. Herb.) ; near Santianne Divide, west side, alt. 850 m., 23 June, 1895, Leiberg 1020 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Granite Station, Kootenai Co., 30 July, 1892, Sandberg, MacDougal & Heller 803 (U. 5. Nat. Herb.) ; near Lewiston, 17 June, 1894, Henderson (U. S. Nat. Herb.) ; Fork of Wood River, alt. 1830 m., 25 July, 1895, Hen- derson 3235 (U. S. Nat. Herb.) ; Twilight Guleh, Owyhee Co., 1916] ` GREENMAN—MONOGRAPH OF SENECIO 135 alt. 1675 m., 23 June, 1911, Macbride 973 (U.S. Nat. Herb. and Mo. Bot. Gard. Herb.); House Creek, Owyhee Co., 29 June, 1912, Nelson Ф Macbride 1808 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.). Utah: Ogden, June, 1871, Coulter (U. S. Nat. Herb.). Nevada: grassy lowlands, Bieroth’s Ranch, McDonald Creek, 2 Aug., 1912, Nelson £ Macbride 2159 (U.S. Nat. Herb.) ; East Humboldt Mountains, alt. 1985 m., U. S. Geol. Exploration of the 40th Parallel, Watson 667 (Gray Herb. and U. S. Nat. Herb.). Washington: eastern Washington, 26 July, 1892, Hender- son (Mo. Bot. Gard. Herb.) ; Ellensburg, 2 June, 1897, Whited 442 (U. 5. Nat. Herb.) ; along streams, Mount Paddo (Adams), 30 June, 1885, Suksdorf (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Faleon Valley, July, 1885, Suksdorf 571 (Gray Herb.); without definite locality, coll. of 1883, Brandegee (Gray Herb.), and coll. of 1889, Vasey 537 (U.S. Nat. Herb.). Oregon: Crow Creek, Wallowa Co., alt. 1295 m., 3 July, 1897, Sheldon 8512 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); on Miller Trail, near Sled Springs, Imnaha National Forest, alt. 1265 m., 12 July, 1907, Jardine 89 (U.S. Nat. Herb.) ; Union Co., coll. of 1879, Cusick 755 (Gray Herb.) ; Minum River, 16 Aug., 1897, Sheldon 8710 (U.S. Nat. Herb.) ; Big Meadows, Des Chutes River, Crook Co., alt. 1370 m., 23 July, 1894, Leiberg 515 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Crater Lake, 10 Aug., 1897, Austin 1610 (U. S. Nat. Herb.); Cascade Mountains, Oregon Boundary Commission, coll. of 1860, Lyall (Gray Herb.) ; Brown's Meadow on Rogue River, alt. 1300 m., 8 July, 1889, Leiberg 4287 (U. S. Nat. Herb.) ; without definite locality, U. S. Exploring Expedition, Wilkes 494 (U. S. Nat. Herb. 48748). California: Goose Lake Valley, July, 1895, Austin 558 (U. S. Nat. Herb. 666887) ; Davis Creek, Modoe Co., Aug., 1894, Austin (Phil. Acad. Nat. Sci. Herb.); Upper Funston Meadows, basin of the Upper Kern River, Tulare Co., alt. 1980 m., July, 1904, Hall Ф Babcock 5569 (Gray Herb.). [Vor. 3 136 ANNALS OF THE MISSOURI BOTANICAL GARDEN Var. flavulus (Greene) Greenm. comb. S. flavulus Greene, Pittonia 4:108. 1900; Rydb. Bull. Torr. Bot. Club 27:185. 1900; Fl. Colo. 397. 1906, in part. S. Balsamitae Nels. in Coulter & A Manual Cent. Rocky Mountains 583. 1909, in part, not Muhl. Stems slender, 1.5 to 4 dm. high; leaves relatively small, the radical and lower stem-leaves ovate-rotund to ovate- oblong, .8 to 4 cm. long, 8 to 20 mm. broad, rounded to acute at the apex, crenate to serrate-dentate, more or less cordate at the base; petioles 1 to 8.5 cm. long; upper stem-leaves petio- late and sublyrate or sessile and laciniate to entire. Distribution: southern Wyoming and Colorado. Specimens examined : Wyoming: river bottoms, Encampment, Carbon Co., alt. 2175 m., 15 June, 1901, Tweedy 4132 (U. S. Nat. Herb.). Colorado: Walden, Larimer Co., 8 July, 1908, Goodding 1494 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Sapinero, alt. 2285 m., 19 June, 1901, Baker 176 (Gray Herb., U. S. Nat. Herb., Greene Herb., and Mo. Pot. Gard. Herb.) ; Black Cañon, alt. 2135 m., 12 June, 1901, Baker 114 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Veta Pass, 15 July, 1896, Shear 3592 (U. S. Nat. Herb.); Arboles, Archuleta Co., 15 June, 1899, Baker 707 (Greene Herb., Berlin Herb., and Mo. Bot. Gard. Herb.), TYPE. 56. S. Burkei Greenm. Ottawa Nat. 25:114. 1911; Ann. Mo. Bot. Gard. 2:626, pl. 20, fig. 1. 1915. An herbaceous perennial, glabrous or nearly so; stems erect, 3 to 9 dm. high, simple or rarely branched, striate; lower leaves petiolate, ovate-oblong, 1 to 7 cm. long, 1 to 3.5 cm. broad, obtuse or rounded at the apex, crenate to serrate-den- tate, cuneate to subtruncate at the base; petioles 2 to 12 cm. long; stem-leaves petiolate and sublyrate to sessile and pin- natisect; inflorescence a terminal few to many-headed corym- bose cyme; heads 10 to 12 mm. high, radiate; involucre cam- panulate, calyculate; bracts of the involucre usually 21, linear- lanceolate, 6 to 8 mm. long, glabrous or floccose-tomentulose, more or less tinged with purple; ray-flowers about 12, rays yellow; disk-flowers numerous; achenes glabrous. 1916] GREENMAN—MONOGRAPH OF SENECIO 137 Distribution: Minnesota to British Columbia. Specimens examined: Minnesota: Itaska Lake, 25 June, 1891, Sandberg 1036 (U. S. Nat. Herb.). Montana: Glacier National Park, 9 July, 1914, Hitchcock 11831 (U.S. Nat. Herb.) ; open ground, shore of Lake MeDon- ald, alt. 950 m., 25 July, 1901, Vreeland (U. S. Nat. Herb. and Geol. Surv. Canada Herb.); Columbia Falls, 1 July, 1894, Williams 68 (U. S. Nat. Herb.). Idaho: wet soil, Kootenai Co., alt. 650 m., Leiberg (Mo. Bot. Gard. Herb.). Rocky Mountains: Grand Saline, ** R. M. E. side", Burke (Gray Herb.), түрЕ; river margins, Silver City, 7 Aug., 1885, Macoun (Gray Herb. and Geol. Surv. Canada Herb. 14772) ; swamps, Kicking Horse Lake, alt. 1540 m., 10 Aug., 1890, Macoun (Geol. Surv. Canada Herb. 14810). Alberta: bank of Bow River, vicinity of Banff, alt. 1370 m., 20 July, 1899, McCalla 2045 (U. S. Nat. Herb.); by the reservoir, Banff, 30 Oct., 1899, Sanson (Geol. Surv. Canada Herb. 22288) ; in vicinity of Banff, July, 1906, Sanson (Geol. Surv. Canada Herb. and Field Mus. Herb.) ; Banff, 28 July, 1904, Farr (Field Mus. Herb.); northern slopes of Crows Nest Pass, 31 July, 1887, Macoun (Geol. Surv. Canada Herb. 22185), in part. British Columbia; open thicket, Spence's Bridge, 31 May, 1889, Macoun (Geol. Surv. Canada Herb. 14811), in part; cleared land at Homer Lake, 19 June, 1905, Shaw 722 (Phil. Acad. Nat. Sci. Herb. and Mo. Bot. Gard. Herb.); Sophie Mountain, between Kettle and Columbia Rivers, 17 July, 1902, Macoun 64990, 64991 (Gray Herb. and Geol. Surv. Caneda Herb.) ; Skagit Valley, alt. 760—925 m., 21 Aug., and 10 July, 1905, Macoun 69358, 69359 (Gray Herb. and Geol. Surv. Can- ada Herb.); shaded banks, mouth of Siliea Creek, Chilliwaek River, 29 June, 1901, Macoun 26685 (Geol. Surv. Canada Herb., Greene Herb., and Gray Herb.), in part; on a bog, Chilliwack Lake, 19 July, 1901, Macoun 26682, 26682a (Geol. Surv. Canada Herb.) ; in a marsh east of Chilliwaek Lake, 25 [VoL. A 138 ANNALS OF THE MISSOURI BOTANICAL GARDEN July, 1901, Macown (Geol. Surv. Canada Herb., Gray Herb., and Mo. Bot. Gard. Herb.) ; Middle Creek, Chilliwack River, 2 Aug., 1901, Macoun 26681 (Geol. Surv. Canada Herb. and Greene Herb.); in thicket by stream at 150-mile house, Cari- boo, 15 July, 1900, Wilson 700 (Geol. Surv. Canada Herb.). 57. S. gaspensis Greenm.' An herbaceous perennial, commonly lightly floccose-tomen- tulose in the axils of the leaves; stems one to several from a common base, erect, 2.5 to 5 dm. high; lower leaves petiolate, broadly ovate to elliptic-lanceolate, .5 to 8 em. long, 1 to 3.5 em. broad, thin, glabrous on both surfaces, or sometimes in the early stages sparingly hairy, especially on the under sur- face, glabrate, rounded to obtuse at the apex, crenate-dentate, abruptly narrowed at the base into a slender petiole 1 to 12 em. in length; inflorescence a terminal several-headed corym- bose cyme; involucre campanulate, calyculate, glabrous or slightly pubescent; bracts of the involucre 13 to 21, linear- lanceolate, 4 to 6 mm. long; ray-flowers 8 to 12, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: Newfoundland, eastern Quebec, and northern Maine. Specimens examined: Newfoundland: meadow and damp talus on hill north of Tilt Cove, Notre Dame Bay, 22 Aug., 1911, Fernald & Wie- gand 6405 (Gray Herb.). Quebec: cold walls of Percé Mountain, Percé, Gaspé Co., 25 July, 1905, Williams, Collins € Fernald (Gray Herb.), iSenecio gaspensis Greenm. sp. nov., herbaceus perennis ad basin et in axillis dm foliorum plerumque albo-floeculoso-tomentulosus ; caulibus erectis 2,5-5 altis; foliis inferioribus petiolatis late ovatis vel elliptico абата 5-8 gis ситни utrinque glabris vel juventa subtu ` sparse pubescentibus — glabratis, ad apicem rotundatis vel obtusis, Ke сен dentatis, ad basin abrupte contractis; pe etioli is gracilibus 1-12 em. longis; inflo- res DH Serer bens omnes term nalibus eapitulis 7-10 mm. altis, radiatis ; involueris eampanulatis едім Ge? parce pubescenti us; bracteis involueri 13-21 lineari-lanceolatis 4-6 mm. longis; floribus femineis 8-12 Ze gen ligulis еи floribus di in numerosis тее quam squamae — achaeniis glabr On eold walls of Pereé Mounta 29 4. А ig , Quebec, 25 July, Ни. та С. Coline & рази пин b.), T А in rand Coupe, "Percé, and B ture Island, Gaspé Co., Aug 1007. Fernald & Collins 1204, 1205 (Gray Herb. >; d Bra 29, 30 July, 1904, Collins, Fernald & Pease (Gray Herb. and U. 8. Nat. Herb)? shaded alluvium, Fort Ke nt, Maine, 6 July, 1904, Fernald (Gray Herb.). 1916] GREENMAN— MONOGRAPH OF SENECIO 139 TYPE; cold northerly calcareous walls of the Grand Coupe, Percé, Gaspé Co., 6 Aug., 1907, Fernald & Collins 1204 (Gray Herb.) ; limestone cliffs, Bonaventure Island, Gaspé Co., 7 Aug., 1907, Fernald & Collins 1205 (Gray Herb.); alluvial thickets, between the Forks and Brilé Brook, Little Casca- pedia, Collins, Fernald £ Pease (Gray Herb. and U. S. Nat. Herb.). Maine: shaded alluvium, Fort Kent, 6 July, 1904, Fernald (Gray Herb.); rocky river flat, Fort Kent, 10 July, 1908, Mackenzie 3418 (U. S. Nat. Herb. 648722). 58. 8. Crawfordii Britt. Torreya 1:21. 1901; Manual 1027. 1901, and ed. 2, 1905; Britton & Brown, Ill. ЕІ. 3:545, fig. 4628. 1913. S. Balsamitae var. Crawfordii (Britt.) Greenm. in Rhodora 10:69. 1908, and in Gray, Manual, ed. 7, 854. 1908. An herbaceous perennial, glabrous throughout or slightly tomentose on the base of the petioles and in the leaf-axils; stems erect, 3 to 6 dm. high; lower leaves long-petiolate, ovate to elliptic-lanceolate, the blades 1 to 8 em. long, 1 to 3.5 em. broad, rounded or obtuse at the apex, erenate to serrate-den- tate, usually abruptly narrowed at the base, glabrous on both surfaces; petioles slender, 2 to 18 em. long; stem-leaves petio- late and more or less lyrate to sessile and incised-serrate; inflorescence a few-headed corymbose cyme; heads about 1 em. high, radiate; involucre campanulate, calyculate; bracts of the involucre 13 to 21, narrowly lanceolate, 6 to 8 mm. long, acute, glabrous, often purplish-tipped; ray-flowers 8 to 12, rays yellow, conspicuous ; disk-flowers numerous; achenes glabrous. Distribution: western New Jersey and southeastern Pennsylvania. Specimens examined: New Jersey: Assinipink Creek, near Trenton and New Brunswick trolley bridge, 28 May, 1904, Brown (Phil. Acad. Nat. Sci. Herb.) ; Abbott’s meadow, below Trenton, 29 May, 1904, Brown (Phil. Acad. Nat. Sci. Herb.) ; Crosswick’s Creek, 29 May, 1904, Williamson (C. S. Williamson Herb.); wet meadow between Springdale and Orchard, Camden Co., 20 May, 1905, Stone (Phil. Acad. Nat. Sci. Herb.). [Vor. 3 140 ANNALS OF THE MISSOURI BOTANICAL GARDEN Pennsylvania: in low wet ground, Tullytown, Bucks Co., 12 Мау, 1894, MacElwee (Phil. Acad. Nat. Sci. Herb.) ; swamp, near Tullytown, 20 May, 1899, Fretz (Phil. Acad. Nat. Sei. Herb.) ; Tullytown, 30 May, 1899, and 22 May, 1900, Craw- ford (Phil. Acad. Nat. Sci. Herb.); Tullytown, May, 1902, Crawford & Brown (Phil. Acad. Nat. Sci. Herb.) ; wet places in bog near Willow Grove, Montgomery Co., 25 May, 1899, MacElwee 326 (Phil. Acad. Nat. Sci. Herb.); Fraser’s bog, east of Willow Grove, 20 May, 1906, Williamson (Phil. Acad. Nat. Sci. Herb.) ; Fraser’s bog, one mile southeast of Willow Grove, 17 and 23 June, 1902, Van Pelt (Phil. Acad. Nat. Sci. Herb.), form; Fraser’s bog, near Bayer’s Corner, 12 May, 1910, Long 3307 (Phil. Acad. Nat. Sci. Herb.); near Phila- delphia, 29 May, 1901, Crawford (Gray Herb.). 59. S.quaerens Greene, Leafl. Bot. Obs. & Crit. 1:214. 1906; Wooton & Standley, Contr. U. S. Nat. Herb. 19:747. 1915. S. prionophyllus Greene, Leafl. Bot. Obs. & Crit. 1:212. 1906, not S. prionophyllus Greene, Ottawa Nat. 15:250. 1902. An herbaceous perennial, glabrous or slightly white-floccose- tomentulose, especially towards the base of the stem, on the petioles, and in the leaf-axils; stems erect, 3 to 6 dm. high, rather leafy at the base, nearly naked above; lower leaves petiolate, subobovate to ovate-oblong, 1 to 8 em. long, 1 to 3 cm. broad, rounded to obtuse at the apex, erenate to subser- rate-dentate ; petioles 1.5 to 14 cm. long; stem-leaves petiolate and sublyrate to sessile and more or less pinnatisect; inflo- rescence a one to many-headed corymbose cyme; heads about 1 em. high, radiate; involucre campanulate, sparingly calyeu- late, slightly tomentulose at the base; bracts of the involucre linear-lanceolate, 5 to 7 mm. long, acute; ray-flowers 8 to 12, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: mountains of New Mexico. Specimens examined : New Mexico: moist places on the west fork of the Gila River, Mogollon Mountains, Socorro Co., alt. about 2285 m., 7 Aug., 1903, Metcalfe 409 (П. S. Nat. Herb. and Mo. Bot. Gard. Herb.), түре; Willow Creek, Mogollon Mountains, 1916] GREENMAN—MONOGRAPH OF SENECIO 141 Socorro Co., 8 Aug., 1900, Wooton (U. S. Nat. Herb.) ; Santa Magdalena Mountains, June, 1881, Vasey (U. S. Nat. Herb.). 60. 8. platylobus Rydb. Bull. Torr. Bot. Club 27:181, pl. 6, fig. 8. 1900. An herbaceous perennial, glabrous or essentially so; stems erect, 2.5 to 4 dm. high, striate; lower leaves petiolate, obovate or broadly oval, including the petiole 3.5 to 14 em. long, 1 to 3 cm. broad, irregularly dentate; stem-leaves petiolate and sublyrate to sessile and pinnately divided into oblong to cuneate, rather conspicuous, rounded or acute lateral divi- sions; inflorescence a many-headed corymbose cyme; heads 8 to 10 mm. high, radiate; involuere campanulate, sparingly calyculate; bracts of the involucre 13 to 21, lanceolate, acute, 5 to 6 mm. long, glabrous, somewhat stramineous; ray-flowers 10 to 12, rays yellow; disk-flowers numerous; achenes gla- brous. Distribution: mountains of Utah. Specimens examined: Utah: Wasatch Mountains, coll. of 1869, Watson 671 (Torrey Herb.), түре; cation bottoms, Provo, Wasatch Moun- tains, 16 June, 1902, Goodding 1115 (Mo. Bot. Gard. Herb.) ; Red Rock Cañon, near Salt Lake City, 11 June, 1905, Rydberg 6064 (U. S. Nat. Herb.) ; rich woods in mountain cañon, Arm- strong 336 (Margaret Armstrong Herb.); Mendon, 17 June, 1898, Mulford 124 (Mo. Bot. Gard. Herb.). There appears to have been some confusion of the material which was distributed by Dr. Watson under his number 671. The specimen in the United States National Herbarium agrees very well partieularly in habit and foliar characters, with typical forms of S. crocatus, but it differs markedly from Wat- son's 671 in the Gray Herbarium (which is apparently S. rubricaulis Greene) and likewise from the specimen bearing the same number in the Torrey Herbarium, namely the type of S. platylobus Rydb. 61. 8. crocatus Rydb. Bull. Torr. Bot. Club 24:299. 1897; ibid. 27:177. 1900, in part; Mem. N. Y. Bot. Gard. 1:446. 1900, at least as to synonymy; Fl. Colo. 396. 1906, in part; Greene, [VoL. 3 142 ANNALS OF THE MISSOURI BOTANICAL GARDEN Pittonia 4:114. 1900; Nelson in Coulter & Nelson, Manual Cent. Roeky Mountains 582. 1909, in part; Daniels, Univ. Mo. Studies, Sei. Ser. 2:252. 1911, in part. S. aureus L.? var. croceus Gray, Proc. Acad. Nat. Sei. Phil. 15:68. 1863, i. e., Hall and Harbour No. 332, in part, and Parry No. 405, not 5. peo DC. S. aureus var. croceus Gray, Syn. Fl. N. Am. 1? :391. 1884, and ed. 2, 1886, in part. S. НИЕТА ив Rydb. Bull. Torr. Bot. Club 27: 176. 1900; Fl. Colo. 396. 1906; Coulter & Nelson, Manual Cent. Rocky Mountains 582. 1909. S. Tracyi Rydb. Bull. Torr. Bot. Club 33:159. 1906; КІ. Colo. 397. 1906. S. pyrrhochrous Greene, Pl. Baker. 3:24. 1901. An herbaceous perennial, glabrous throughout or slightly tomentulose in the axils of the braets of the inflorescence; stems erect, 1 to 7.5 dm. high from a rather stout rootstock, striate; lower leaves petiolate, oblong-ovate, subcordate to abruptly contracted at the base, rounded, obtuse or submucro- nate at the apex, entire to somewhat crenate-dentate; petioles 1 to 12 em. long; stem-leaves petiolate and sublyrate to ses- sile and semiamplexicaul; inflorescence a few to many-headed corymbose cyme; heads in anthesis 8 to 10 mm. high, radiate; involuere eampanulate, sparingly calyculate; bracts of the in- voluere (13-) 21, linear-lanceolate, 5.5 to 8 mm. long, acute, glabrous, more or less tinged with purple; ray-flowers 10 to 12, rays orange-red or saffron-colored varying to yellow; disk- flowers numerous; achenes glabrous. Distribution: mountains of Colorado and Utah. Specimens examined: Colorado: Medicine Bow Mountains, 3 Aug., 1891, Cran- дай (U. S. Nat. Herb.) ; Rocky Mountain Flora, Lat. 39-41°, coll. of 1862, Hall € Harbour 332 (Gray Herb.), in part, TYPE; Middle Park, coll. of 1862, Parry 405 (Gray Herb., Mo. Bot. Gard. Herb., and U. S. Nat. Herb. 349244) ; without definite locality, Wolf 4 Rothrock 581 (Gray Herb., U. S. Nat. Herb., and Phil. Aead. Nat. Sei. Herb.); Gray's Peak, July, 1888, 1916] GREENMAN—MONOGRAPH OF SENECIO 143 Eastwood (U. S. Nat. Herb.); Park Co., Williamson (C. 8. Williamson Herb.) ; Dickey, below Breckenridge, 2 Sept., 1885, ex Herb. Fritchey (Mo. Bot. Gard. Herb.) ; near Breckenridge, coll. of 1892, Wislizenus 1086, and Aug., 1901, Mackenzie 211 (Mo. Bot. Gard. Herb.) ; Mt. Baldy, 15 July, 1906, Anderson (Mo. Bot. Gard. Herb.) ; vicinity of Twin Lakes, 2 Aug., 1873, Coulter (Phil. Acad. Nat. Sci. Herb.) ; Mt. Lincoln, alt. 3650 m., 9 July, 1873, Coulter (U. S. Nat. Herb.) ; Weston's Pass, 18 July, 1873, Coulter (U. S. Nat. Herb.) ; damp meadows, Elk Mountains, coll. of 1881, Brandegee (Mo. Bot. Gard. Herb.) ; Jack's Cabin, Gunnison Co., alt. 2520 m., 26 Aug., 1901, Baker 612 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; vicinity of Mount Carbon, Gunnison Co., alt. 2750 m., 6 July, 1910, Eggleston 5866 (U. S. Nat. Herb.); Marshall Pass, 27 July, 1896, Shear 5162 (U. S. Nat. Herb.); Sargents, Sagu- ache Co., alt. 2580 m., 5 July, 1901, Baker 348 (Gray Herb., U. S. Nat. Herb., Greene Herb., and Mo. Bot. Gard. Herb.) ; Silverton, 5 Aug., 1897, Shear 4900 (U. S. Nat. Herb.) ; Ha- mor's Lake, north of Durango, alt. 2740 m., 24 July, 1898, Baker, Earle & Tracy 625 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.). Utah: Wasatch Mountains, alt. 1525 m., May, 1869, Watson 671 (U. S. Nat. Herb.) ; Fish Lake, National Forest, alt. 2995 m., 19 July, 1913, Arriveé (U. S. Nat. Herb.). Var. Wolfii Greenm. comb. nov. S. Wolfii Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. Stems slender, 2 to 3 dm. high; lower leaves ovate-rotund to ovate-oblong, 7 to 20 mm. long, 5 to 13 mm. broad, rounded or obtuse at the apex, entire, subeordate to cuneate at the base. Similar to the species but somewhat more slender and with smaller leaves. Distribution: mountains of Colorado. Specimens examined: Colorado: South Park, Lieut. Wheeler's Expedition, 1873, Wolf € Rothrock 582, 586 (Gray Herb. and U. S. Nat. Herb.), TYPE; meadows, South Cottonwood Gulch, Chaffee Co., alt. 3050 m., 9 July, 1892, Sheldon 481 (U. S. Nat. Herb.). [Vor. 3 144 ANNALS OF THE MISSOURI BOTANICAL GARDEN 62. 6. aquariensis Greenm.' An herbaceous perennial, glabrous or slightly tomentulose in the inflorescence; stems erect, solitary or closely cespitose, 2 to 3.5 dm. high; lower leaves petiolate, ovate to oblong- lanceolate, 1 to 7 em. long, D to 2 em. broad, obtuse to acute, entire, glabrous on both surfaces; petioles 1 to 6 cm. long; stem-leaves petiolate and sublyrate to sessile, semiamplex- ieaul and more or less pinnatisect; inflorescence a terminal rather close corymbose cyme; heads numerous, 8 to 10 mm. high, sparingly calyculate, radiate; involucre campanulate ; bracts of the involuere 13 (—21), jawas lat, 5 to 6 mm. long, acute, glabrous; ray-flowers 10 to 12, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: high plateaus of Utah. Specimens examined: Utah: Aquarius Plateau, alt. 3050 m., 5 Aug., 1875, Ward 505 (Mo. Bot. Gard. Herb., Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sei. Herb., and Field Mus. Herb.), TYPE; Bear River Valley, coll. of 1877, Palmer 267V5 (Gray Herb., U. S. Nat. Herb. 782529 in part, and Mo. Bot. Gard Herb.). 63. S. dimorphophyllus Greene, Pittonia 4:109. 1900; Rydb. Bull. Torr. Bot. Club 27:178. 1900; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. S. heterodoxus Greene ex Rydb. Bull Torr. Bot. Club 27:178. 1900; КІ. Colo. 396. 1906, in synonymy. S. aureus var. croceus Gray, Droe, Acad. Nat. Sci. Phil. 15:68. 1863, i. e., Hall & Harbour No. 332, in part; Gray, Syn. Fl. N. Am. 12:391. 1884, and ed. 2, 1886, іп part; Porter & 'Senecio aquariensis Greenm. ov. herbaceus perennis glabrus vel in inflorescentiis leviter tomentosus; e алы ereeti is 2-3 dm. altis solitariis vel dense eaespitosis; foliis inferioribus petiolatis "€ Na oblongo-lanceolatis 1-7 em 1 . latis obtu o ad [pee abrupte cune atis vel raro sub E. peti olis 1-6 ¢ m. longis; foliis su- perioribus petiolatis et sublyratis vel — plus minusve e pinnatiseetis semi- amplexicaulibusque; infloreseentiis terminalibus conferte corymboso-cymosis ; capitulis numerosis 8-10 n altis e ea alyeulatis radiatis; involucris cam- panulatis erg pasce 3 (-21) laneeo olatis 5-6 mm. ongis aeutis glabris; floribus femineis 10- 12, gulis flavibus; floribus disci numerosis; achaeniis Pome —Colleeted on the Aquarius Plateau, m 3050 m an, а 1875, m 505 (Mo. Bot. Gard. ч, St hw rb., U. 8. Nat. Herb., Phil. б Ge "Field Mus. b); B r River Valley, SCH of 1877, “Palmer 267% (Gray Herb., U. 8. Nat. Herb. 782529 in part, and Mo. Bot. Gard. bh. 1916] GREENMAN—MONOGRAPH OF SENECIO 145 Coulter, Syn. Fl. Colo. 82. 1874, in part; Coulter, Manual Rocky Mountain Region 211. 1885, in part. S. crocatus Rydb. Mem. N. Y. Bot. Gard. 1:446. 1900, in part; Bull. Torr. Bot. Club 27:177, pl. 5, fig. 13. 1900, as to deseription, illustration, and most of specimens cited, not as to synonymy ; Fl. Colo. 396. 1906, mainly ; Daniels, Fl. Boulder 202. 1911, mainly; Nelson in Coulter & Nelson, Manual Cent. Rocky Mountains 582. 1909, in part. An herbaceous perennial, glabrous or essentially so throughout; stems one to several from a common base, erect or ascending, 1 to З dm. high; lower leaves ovate, subobovate, broadly spatulate or somewhat oblong-lanceolate, 1 to 4 em. long, .5 to 2.5 em. broad, rounded to obtuse at the apex, entire to crenate, narrowed at the base into a winged petiole equal- ling or exceeding the blade; stem-leaves mostly sessile, oblong- lanceolate to triangular-ovate, frequently conspicuously di- lated at the base and amplexicaul; inflorescence a few to sev- eral-headed terminal corymbose cyme; heads about 1 cm. nigh, radiate ; involucre campanulate, calyculate; bracts of the involuere about 21, linear-lanceolate, 6 to 7 mm. long, acumi- nate, acute, glabrous or slightly tomentulose, frequently red- dish-tipped; ray-flowers 10 to 12, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: mountains of Wyoming and Colorado. Specimens examined: Wyoming: La Plata Mines, 21 Aug., 1895, A. Nelson 1769 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; wet, subalpine woods, Nash’s Fork, Albany Co., 15 Aug., 1908, A. Nelson 9148 (Mo. Bot. Gard. Herb.) ; grassy ground, below snow, Bridges Peak, Carbon Co., 24 Aug., 1903, Goodding 1980 (U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.). Colorado: alpine meadows, summit of North Park Range, Larimer Co., 10 Aug., 1908, Goodding 1820 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); about timberline, above Berth- oud's Pass, 14 Sept., 1874, G. Engelmann (Mo. Bot. Gard. Herb.); timberline, Long's Peak, 5 Aug., 1886, Letterman 10 [VoL. 3 146 ANNALS OF THE MISSOURI BOTANICAL GARDEN (Mo. Bot. Gard. Herb.); foot of Kelso Mountain, near Tor- rey’s Peak, 14 Aug., 1885, Letterman (Mo. Bot. Gard. Herb.) ; Front Range, alt. 3500 m., 6 July, 1896, Crandall (Mo. Bot. Gard. Herb.); Grays Peak, above timberline, 31 Aug., 1884, B. H. Smith (Phil. Acad. Nat. Sci. Herb.) ; Grays Peak, alt. 3650-3800 m., 15 Aug., 1885, Letterman (Mo. Bot. Gard. Herb. and Field Mus. Herb.) ; Powell's Colorado Expl. Exp., 1868, Lat. 40-41°, Vasey 340B (Gray Herb. and Mo. Bot. Gard. Herb.) ; Lat. 89—41°, coll. of 1862, Hall £ Harbour 332 (U.S. Nat. Herb. 48721, Phil. Acad. Nat. Sei. Herb., and Field Mus. Herb. 314668); Lat. 39—41°, coll. of 1862, Hall £ Harbour 331 (U. S. Nat. Herb. 48766), in part, and Hall £ Harbour 115 (Gray Herb.), in part; without definite locality, coll. of 1871, Brandegee 132 (Mo. Bot. Gard. Herb.) ; Breckenridge, coll. of 1892, Wislizenus 1067 (Мо. Bot. Gard. Herb.) ; Mt. Parry, coll. of 1872, Gray (Gray Herb.) ; Golden City, 12 July, 1871, Greene 528 (Gray Herb.) ; Golden, 13 July, 1885, Letterman (Mo. Bot. Gard. Herb.) ; Peak Valley, 21 Aug., 1901, F. E. «& E. S. Clements 485 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Cameron Pass, alt. 3500 m., 14 July, 1896, Baker (Mo. Bot. Gard. Herb.) ; meadows, South Cottonwood Guleh, Chaffee Co., alt. 3050 m., 9 July, 1892, Sheldon 100 (U. S. Nat. Herb.) ; Gunnison Co., July, 1889, Eastwood (U. S. Nat. Herb.) ; Mineral Point, July, 1887, Hempton (Mo. Bot. Gard. Herb.); near Pagosa Peak, Mineral Co., alt. 3200 m., 6 Aug., 1899, Baker 705 (Gray Herb., Berlin Herb, Greene Herb., and Mo. Bot. Gard. Herb.), түре; head of Vallecito, alt. 3000-3800 m., 2 Sept., 1903, Knowlton 3 (U. S. Nat. Herb.) ; La Plata Mountains, alt. 3650 m., 15 July, 1896, Tweedy 537 (U. S. Nat. Herb.) ; Little Kate Mine, La Plata Mountains, alt. 3500 m., 14 July, 1898, Baker, Earle £ Tracy 569 (Gray Herb., U. S. Nat. Herb., Greene Herb., and Mo. Bot. Gard. Herb.), түре of S. heterodoxus Greene; Farnham, 10 July, 1891, E. C. Smith (Mo. Bot. Gard. Herb., 64. S. Farriae Greenm. Bot. Gaz. 42:147. 1906; Contr. Bot. Lab. Univ. Penn. 3:74. 1907 ; Ottawa Nat. 25:115. 1911. An herbaceous perennial, glabrous except for a persistent white tomentum in the axils of the leaves; stems erect or as- 1916] GREENMAN—MONOGRAPH OF SENECIO 147 cending, 1 to 3 dm. high, simple or branched from near the base; lower leaves ovate to slightly obovate, the blade 1 to 4 em. long, 1 to 2.5 em. broad, rounded at the apex, erenate to subentire, contracted at the base into a narrowly winged petiole equalling or exceeding the blade; stem-leaves petiolate and sublyrate to sessile and somewhat irregularly pinnatifid, the uppermost leaves reduced to mere bracts; heads about 1 em. high, radiate; involucre campanulate, slightly calyculate, sparingly tomentulose at the base; bracts of the involucre about 21, linear-lanceolate, 7 to 8 mm. long, frequently red- dish-tipped; ray-flowers 10 to 14, rays yellow; disk-flowers numerous, 50 to 60; achenes glabrous. Distribution: mountains of Alberta to Washington. Specimens examined: Alberta: near Banff, alt. 1500 m., 8 June, 1904, Farr (Univ. Penn. Herb. and Field Mus. Herb.), түре; vicinity of Banff, July, 1906, Sanson 81260 (Geol. Surv. Canada Herb. and Mo. Bot. Gard. Herb.) ; vicinity of Basin, near Banff, alt. 1400 m., 8 and 18 June, 1906, Brown 20 (Phil. Acad. Nat. Sci. Herb.) ; in deep moss in stream-bed below warm sulphur spring, vi- einity of Banff, alt. 1370 m., 15 June, 1899, McCalla 2049 (U. S. Nat. Herb.) ; Sulphur Springs, Banff, alt. 1415 m., 11 June, 1906, Butters £ Rosendahl 1324 (Field Mus. Herb.) ; Crows Nest Lake, alt. 1385 m., 9 July, 1883, Dawson (Geol. Surv. Canada Herb. 14800), in part; Devil's Head Lake, alt. 1385 m., 13 July, 1899, Sanson (Geol. Surv. Canada Herb. 22125) ; erossing of MeLeod's River, 19 June, 1898, Spreadborough (Geol. Surv. Canada Herb. 19725); in grass along Bragg's Creek, Elbow River, 26 June, 1897, Macoun (Geol. Surv. Can- ada Herb. 22784) ; damp places, Red Deer, coll. of 1895, Gaetz (Geol. Surv. Canada Herb. 11622). Washington: on rocky bar of Columbia River at Wenatchee, 25 Мау, 1899, Whited 1096 (U. S. Nat. Herb.). 65. 8. Hartianus Heller, Bull. Torr. Bot. Club 26:622. 1899. S. flavulus Wooton & Standley, Contr. U. S. Nat. Herb. 19:747. 1915, in part, not Greene. An herbaceous perennial, at first somewhat white-tomentu- lose, later more or less glabrate; stems erect, 2 to 5 dm. high, [Vor. 8 148 ANNALS OF THE MISSOURI BOTANICAL GARDEN lower leaves petiolate, ovate-rotund, ovate-oblong to subobo- vate, 1 to 3.5 em. long, .5 to 2 em. broad, rounded to obtuse at the apex, minutely erenate to serrulate, subeordate to abruptly contracted to a cuneate base; petioles 1 to 6.5 em. long; inflo- rescence a few to many-headed corymbose всуше; heads 6 to 10 mm. high, radiate; involuere campanulate, sparingly calyeu- late, tomentulose; bracts of the involucre 13 to 21, linear, 4 to 6 mm. long; ray-flowers about 12, rays pale yellow; disk- flowers numerous; achenes glabrous. Distribution: mountains of New Mexico and Arizona. Specimens examined: New Mexico: Winsor’s Ranch, Pecos River National Forest, alt. 2500 m., 30 June, and 3 July, 1908, Standley 4058, 4061, 4165 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; marsh, Kingston, Sierra Co., alt. 2000 m., 25 May, 1904, Met- calfe 931 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.). Arizona: vicinity of Flagstaff, alt. 2135 m., 5 July, 1898, MacDougal 230 (Gray Herb. and Phil. Acad. Nat. Sci. Herb.), TYPE; San Francisco Mountains, 7 July, 1892, Тоитеу 663 (Gray Herb. and U. S. Nat. Herb.) ; San Francisco Moun- tains, 21 Aug., 1889, Knowlton 64 (U. S. Nat. Herb.) ; Mt. Agassiz, coll. of 1884, Lemmon 38283 (Gray Herb.); near Kendrick Mountains, alt. 2000 m., 7 July, 1901, Leiberg 5663 (U. S. Nat. Herb.) ; Cooley's Ranch, Navajo Co., 1 July, 1912, Goodding 1108 (U. S. Nat. Herb.). 66. S. plattensis Nutt. Trans. Am. Phil. Soc. N. S. 7:413. 1841; Britton & Brown, Ill. Fl. 3:478, fig. 4039. 1898, and ed. 2, 043, fig. 4623. 1913; Heller, Cat. N. Am. Pl. 146. 1898, and ed. 2, 230. 1900; Rydb. Mem. N. Y. Bot. Gard. 1:445. 1900, and Bull. Torr. Bot. Club 27:185, pl. 6, fig. 14. 1900; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Britton, Manual 1026. 1901, and ed. 2, 1905; Rydb. Fl. Colo. 396. 1906; Coulter & Nelson, Manual Cent. Rocky Mountains 581. 1909; Daniels, Univ. Mo. Studies, Sci. Ser. 2:251. 1911. S. balsamitae Torr. in Nicollet’s Report, App. B, 153 [237]. 1843, not Muhl. 1916] GREENMAN— MONOGRAPH OF SENECIO 149 S. aureus var. Balsamitae Torr. & Gray, Fl. N. Am. 2:442. 1843, in part; Gray in Boston Jour. Nat. Hist. 6:231. 1857; Gray, Syn. Fl. N. Am. 1? :391. 1884, and ed. 2, 1886, in part. S. camporum Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. S. pseudotomentosus Mack. & Bush, Trans. Acad. Sci. St. Louis 12:88, pl. 17. 1902. An herbaceous perennial, usually more or less persistently white-floecose-tomentulose, rarely glabrous throughout ; stems erect, one to several from a common base, 1 to 4 dm. high; leaves variable, the lower ovate-oblong to lanceolate or some- what oblanceolate, 1 to 8 em. long, .5 to 4 em. broad, rounded to obtuse at the apex, crenate to serrate-dentate, subcordate to gradually narrowed at the base; petioles 1 to 15 em. long; stem-leaves petiolate and sublyrate to sessile and very ir- regularly pinnatisect; inflorescence a terminal corymbose cyme; heads usually numerous, 8 to 10 mm. high, radiate; involuere eampanulate, calyculate; bracts of the involuere 13 (21), linear-lanceolate, 5 to 6 mm. long, glabrous or slightly tomentulose, penicillate; ray-flowers 10 to 12, rays yellow; disk-flowers numerous; achenes usually hispidulous along the angles, sometimes glabrous. Distribution: southwestern Ontario to Saskatchewan and eastern Montana, south to Louisiana and Texas. Specimens examined: Ontario: on sand dunes, Port Franks, Lambton Co., 24 May, 1906, and 9 Aug. 1907, Dodge 298, 108 (U. S. Nat. Herb.) ; Camlachie, 18 June, 1901, Macoun (Gray Herb. and U. S. Nat. Herb.) ; northeast of Sarnia, 5 June, 1897, Dodge 297 (U. S. Nat. Herb.). Manitoba: Stony Mountain, 14 June, 1887, Fowler (Mo. Bot. Gard. Herb.) ; open places south of Sewell, 12 June, 1876, Macoun 12232 (Geol. Surv. Canada Herb.) ; gravelly or rocky places, Fort Ellice, 20 June, 1879, Macoun 14799 (Geol. Surv. Canada Herb.). Saskatchewan: Long Lake (Last Mountain Lake), 6 July, 1879, Macoun 48 (Gray Herb.). [Vor. 3 150 ANNALS OF THE MISSOURI BOTANICAL GARDEN Michigan: dry fields and dryish woods, near Rochester, 2 June, 1912, and 25 May, 1913, Farwell 2616, 3383, 3394 (Mo. Bot. Gard. Herb.); sterile hills, and in marl beds, Parkdale Farm, 2 June, 1912, 25 May, and 8 June, 1913, and 30 May, 1914, Farwell 2608, 3414, 9443, 3655 (Mo. Bot. Gard. Herb.) ; dry soil in oak openings, Grand Rapids, 12 June, 1893, Cole (Gray Herb.); without definite locality, coll. of 1902, H. L. Clark (U. S. Nat. Herb. 413298). Indiana: Notre Dame, 2 June, 1908, Niewwland, and 14 May, 1913, Nieuwland 11079 (Mo. Bot. Gard. Herb.). Wiseonsin: Fort Howard, on beaver dam, 15 June, 1878, Schuette (U. S. Nat. Herb. and Field Mus. Herb.). Ilinois: gravelly bluffs, Ringwood, coll. of 1860, Vasey (Gray Herb.) ; moist prairie, near Wady Petra, 20 May, 1896, Chase (Mo. Bot. Gard. Herb.) ; dry gravelly soil, Peoria, May and June, 1904, McDonald (Gray Herb.) ; vicinity of Oquawka, Patterson (Gray Herb. and Field Mus. Herb.); Mississippi River valley opposite St. Louis, 13 May, 1874, Eggert (Gray Herb., U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.) ; dry hills, French Village, 14 May, 1875, 14 May, 1878, and 8 May, 1892, Eggert (Mo. Bot. Gard. Herb.) ; French Vil- lage, 23 April, 1878, Eggert (U. S. Nat. Herb. and Mo. Pot. Gard. Herb.) ; French Village, 17 May, 1894, Glatfelter (Mo. Bot. Gard. Herb.); bluffs, near Belleville, April, 1834, (?) Engelmann 592 (Mo. Bot. Gard. Herb.) ; without definite lo- cality, Brendel (Berlin Herb. and Gray Herb.). Missouri: Windsor Springs, 26 April, 1890, Hitchcock (Mo. Bot. Gard. Herb.) ; Cliff Cave, 3 May, 1901, J. H. Kel- logg (Mo. Bot. Gard. Herb.); Kimmswiek, 10 May, 1885, Wislizenus 226 (Mo. Bot. Gard. Herb.), in part; Kimmswick, 9 May, 1915, Drushel 1262 (Mo. Bot. Gard. Herb. and J. A. Drushel Herb.) ; sandy rocks, near Crystal City, 20 May, 1887, Eggert (Mo. Bot. Gard. Herb.) ; Potosi, 3 June, 1892, Dewart 96 (Mo. Bot. Gard. Herb.) ; dry ground, near Bismarck, 30 April, 1893, Eggert (Mo. Bot. Gard. Herb.) ; Monteer, 24 May, 1900, 13 May, 1901, 2 May, 1902, and 27 April, 1907, Bush 711, 455, 1487 and 4338 (Mo. Bot. Gard. Herb.) ; Grandin, 5 May, 1901, Bush 344 (Mo. Bot. Gard. Herb.) ; Grandin, 6 May, 1905, 1916] GREENMAN— MONOGRAPH OF SENECIO 151 Bush 2706, 27064 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Butler Co., 1 May, 1905, Bush 2558 (Mo. Bot. Gard. Herb.) ; Pleasant Grove, 20 Мау, 1900, Bush 336 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Pleasant Grove, 21 May, 1900, Bush 363 (Mo. Bot. Gard. Herb.); Jerome, 28 April, 1914, J. H. Kellogg 453 (Mo. Bot. Gard. Herb.) ; Springfield, 28 April, 1888, Weller 683 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Watson, 1 June, 1894, Bush 180 (Gray Herb. and Mo. Bot. Gard. Herb.); prairies, Lee’s Summit, 9 May, 1897, Bush 349 (Mo. Bot. Gard. Herb.); in woods, Courtney, 24 May, 1903, Bush 1830 (Mo. Bot. Gard. Herb.); wooded hills, Courtney, 12 May, 1913, Bush 6995 (Mo. Bot. Gard. Herb.); rocky woods, Dodson, 5 May, 1914, Bush 7105 (Mo. Bot. Gard. Herb.); sandy hills and low prairies, Cass Co., May-June, 1885, and April, 1871, Broadhead (Mo. Bot. Gard. Herb.); prairies, Webb City, 4 May, 1902, E. J. Palmer 314 (Mo. Bot. Gard. Herb.); high prairies, Carthage, 14 May, 1911, and 4 May, 1913, E. J. Pal- mer 3373, 3383, 3942 (Mo. Bot. Gard. Herb.); high sandy prairies, Alba, 7 May, 1914, Е. 7. Palmer 5518 (Mo. Bot. Gard. Herb.) ; Eagle Rock, 14 June, 1897, Bush 182 (Mo. Bot. Gard. Herb.) ; McDonald Co., 22 April, 1891, Bush (Мо. Bot. Gard. Herb.). Arkansas: Corning, May, 1884, Letterman (Berlin Herb., fragment in Gray Herb.) ; ‘‘Monark,’’ 2 Мау, 1905, Bush 2592 (Mo. Bot. Gard. Herb.). Louisiana: west of Meriden, 15 April, 1901, Canby, Sar- gent, Trelease & Bush 144 (Phil. Acad. Nat. Sei. Herb.); without definite locality, Bradbury (Phil. Acad. Nat. Sci. Herb.) ; ‘‘Upper Louisiana,” Nuttall (Phil. Acad. Nat. Sci. Herb.), (?) түре; without definite locality, Hale, ex Short Herb. (Phil. Acad. Nat. Sci. Herb.). Minnesota: Houston Co., May, 1912, Freiberg (Mo. Bot. Gard. Herb.) ; railroad banks, Perham, Ottertail Co., 20 July, 1912, Chandonnet (Mo. Bot. Gard. Herb.). Iowa: Fayette Co. З June, 1894, Fink 10 (U.S. Nat. Herb.); Iowa City, Hitchcock (Mo. Bot. Gard. Herb.) ; dry [Vor. 3 152 ANNALS OF THE MISSOURI BOTANICAL GARDEN hillsides, Lee Co., 19 May, 1914, Rev. John Davis 2371, 3822 (Mo. Bot. Gard. Herb.) ; low ground on prairies, Armstrong, 14 June, 1883, and 5 June, 1898, Cratty (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Burnside, 1 June, 1899, Somes C. 3014 (U. S. Nat. Herb.) ; wet meadows, Decatur Co., 28 May, 1898, Anderson (Mo. Bot. Gard. Herb.); dry hills, near Council Bluffs, Nicollet’s North-Western Expedition, 16 May, 1839, Geyer 98 (Phil. Acad. Nat. Sci. Herb., Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; without definite locality, 9 June, 1875, Arthur (Mo. Bot. Gard. Herb.). North Dakota: Conway, 2 July, 1896, Brannon 193 (Mo. Bot. Gard. Herb.) ; dry prairie, Grand Forks, 12 June, 1896, Brannon 232 (Mo. Bot. Gard. Herb.) ; on level prairie, Rogers, 10 June, 1912, Bergman 1696 (Mo. Bot. Gard. Herb.) ; along bottom of open ravine, Adrian, 27 June, 1912, Bergman 1808 (Mo. Bot. Gard. Herb.); wet prairie, Leeds, 11 June, 1899, Lunell (Gray Herb.). South Dakota: prairies, ‘‘Oakwood,’’ 23 May, 1902, А. G. J. (Mo. Bot. Gard. Herb.); Brookings, coll. of 1892, Walliams (U. S. Nat. Herb.) ; Brookings, 20 June, 1893, Thornber (Mo. Bot. Gard. Herb.) ; Brookings, June, 1905, White (Mo. Bot. Gard. Herb.) ; Sioux Falls, 18 May, 1894, Thornber (Mo. Bot. Gard. Herb.); Fort Pierre to Yellowstone River, Raynolds’ Expedition to the Headwaters of the Missouri and Yellow- stone Rivers, July, 1859, Hayden (Mo. Bot. Gard. Herb.) ; grassy draws on plains, Washabaugh Co., 22 May, 1914, Over 3088 (U. S. Nat. Herb.); Black Hills, near Fort Meade, 2] June, 1887, Forwood 231 (U. S. Nat. Herb.) ; Custer, 16 July, 1892, Rydberg 827 (Gray Herb.), in part; Hot Springs, 18 June, 1892, Rydberg 828 (Gray Herb. and U. S. Nat. Herb.) ; Mayo, 18 June, 1914, Over 1895 (U. S. Nat. Herb.). Montana: Miles City, 26 May, 1902, Blankenship (Gray Herb.). Nebraska: one hundred miles above Council Bluffs, 4 June, 1853, Hayden (Mo. Bot. Gard. Herb.) ; Mauvaises Terres, 12 July, 1853, Hayden (Mo. Bot. Gard. Herb.) ; Fremont, 3 June, 1893, Schenck 6 (Mo. Bot. Gard. Herb.) ; prairies near Lincoln, 1916] GREENMAN—MONOGRAPH OF SENECIO 153 June, 1887, Webber, also 10 and 12 May, 1900, Hedgcock (Mo. Bot. Gard. Herb.) ; Minden, 11 June, 1907, Hapeman (Mo. Bot. Gard. Herb.) ; Big Blue and Little Blue Rivers to Big Platte River, June, 1849, Fendler 71 (Gray Herb.); Republican Valley, Franklin Co., 15 May, 1893, Laybourn (Mo. Bot. Gard. Herb.); Hershey, 20 May, 1903, Mell 46 (U. S. Nat. Herb.) ; along streams, Halsey, 27 May, 1903, Mell & Knopff (Mo. Bot. Gard. Herb.); Platte bottom, Kearney Со., 15 June, 1891, also Cheyenne Co., 3 Aug., 1891, Rydberg 211 (U. 5. Nat. Herb.); Nebraska (?), Stevens' Pacifie Railway Expedition (U. S. Nat. Herb. 48724, 48726). Colorado: near Evans and Greeley, colls. of June, 1907, 1908, and 1909, E. L. Johnston 439, 441, 443, 445, 460, 546 (Mo. Bot. Gard. Herb.); New Windsor, Weld Co., 4 June, 1901, Osterhout (U. S. Nat. Herb. and Phil. Acad. Nat. Sci. Herb.) ; Fort Lupton, in Platte River bed, 19 May, 1 and 15 June, 1913, Е. L. Johnston 884, 872, 868 (U. S. Nat. Herb.) ; river flats, Fort Collins, alt. 1525 m., 17 May, 1895, Crandall 281 (0. 5. Nat. Herb. and Mo. Bot. Gard. Herb.) ; cañon of the Cache-la- Poudre, 2 June, 1891, Crandall (U. S. Nat. Herb.) ; Denver, Lieut. Wheeler's Expedition, June, 1873, Wolf 556 (U. 5. Nat. Herb.) ; low meadows on Clear Creek, 23 May, 1870, Greene 226 (Gray Herb.) ; near Breckenridge, 22 July, 1906, Anderson (Mo. Bot. Gard. Herb.); Horsetooth Guleh, 28 May, 1898, Crandall 3070 (U. S. Nat. Herb.) ; wet meadow valley, 20 June, 1873, Brandegee (Mo. Bot. Gard. Herb.). Kansas: Manhattan, coll. of 1884, Kellerman 8 (Gray Herb.); Manhattan, 15 May, 1887, Kellerman (Mo. Bot. Gard. Herb.); Manhattan, 6 May, 1891, Fritz (U. S. Nat. Herb.) ; stony hills, Riley Co., 9 May, 1895, and coll. of 1896, Norton 303 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Emporia, 29 April, 1890, Tyler (Mo. Bot. Gard. Herb.) ; Cowley, White (Mo. Bot. Gard. Herb.) ; dry prairies, near Osborne City, 14 May, 1894, Shear 28 (Gray Herb., U. 8. Nat. Herb., and Mo. Bot. Gard. Herb.). Oklahoma: Huntsville, Kingfisher Co., 29 April, 1896, Blankenship (Gray Herb., U. S. Nat. Herb., and Mo. Bot. [Vor. 3 154 ANNALS OF THE MISSOURI BOTANICAL GARDEN Gard. Herb.) ; Sapulpa, 29 and 30 April, 1895, Bush 1249, 975 (Mo. Bot. Gard. Herb.); between Fort Cobb and Fort Arbuckle, Palmer 461 (U. S. Nat. Herb.) ; without definite locality, 16 April, 1898, Waugh 275 (Mo. Bot. Gard. Herb.) ; without locality, Stevens 34, 50.24, 120, 134, 146, 163, 179.6 А, 210, 233, 286V5, 301V5, 420H, 469 (Geo. W. Stevens Herb.). Texas: rich woods, Dallas, March, 1882, Reverchon 556 (Mo. Bot. Gard. Herb.) ; on prairie, Dallas, 15 April, 1900, Bush 606 (U.S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; rocky prairies, Fort Worth, 21 April, 1902, Reverchon (Mo. Bot. Gard. Herb.) ; hillsides, Polytechnic, 5 March, 1913, Ruth 54 (Mo. Bot. Gard. Herb.) ; Limestone Co., March, 1878, Joor (U. S. Nat. Herb.) ; College Station, Brazos Co., Ness 2083 (Mo. Bot. Gard. Herb.) ; sandy prairies, Terrell, 6 April, 1903, Reverchon 3964 (Gray Herb. and Mo. Bot. Gard. Herb.); Round-top Mountain, Comanche Co., 9 May, 1900, Eggert (Mo. Bot. Gard. Herb.) ; Georgetown, March, 1890, Bodin 53 (U. S. Nat. Herb); Gillespie Co., ex Herb. Jermy (Mo. Bot. Gard. Herb.); New Braunfels, April, 1851, Lindheimer (Mo. Bot. Gard. Herb.) ; in patches on rocky high plains, Upper Guade- loupe, coll. of March, 1846, Lindheimer 445 (Gray Herb., U. 8. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.). 67. S. Willingii Greenm. Ottawa Nat. 25:117. 1911. An herbaceous perennial, somewhat glaucous; stems erect, 2.5 to 3 dm. high, glabrous, striate, leafy; leaves oblong- lanceolate, 3 to 12 em. long, .7 to 2 em. broad, erenate-serrate to pinnately divided into oblong, entire or obtusely dentate lobes obtuse to rounded at the apex, in the early stages Йоссове- tomentulose along the midrib and lateral nerves beneath and on the margins of the petioles, later glabrate; inflorescence a terminal, rather dense, corymbose eyme; heads 8 to 10 mm. high, radiate; involuere eampanulate, calyculate, sparingly floccose-tomentulose, glabrate; bracts of the involucre about 21, linear-lanceolate, 6 to 7 mm. long, acute; ray-flowers about 12, rays yellow; disk-flowers numerous, 60 to 70; achenes glabrous. 1916] GREENMAN—MONOGRAPH OF SENECIO 155 Distribution: Manitoba to Alberta. Specimens examined: Manitoba: gravelly soil, Ninga, 1 June, 1908, Hales 24 (Geol. Surv. Canada Herb.). Alberta: near Olds, Aug., 1894, Walling (Geol. Surv. Can- ada Herb. 14843, 6063, fragment and photograph in Field Mus. Herb.), TYPE. 68. S. Smallii Britt. Mem. Torr. Bot. Club 4:132. 1894; Britton & Brown, Ill. Fl. 3:479, fig. 4044. 1898, and ed. 2, 546, fig. 4630. 1913; Heller, Cat. N. Am. Pl. 147. 1898, and ed. 2, 231. 1900; Britton, Manual 1028. 1901, and ed. 2, 1905; Small, Fl. Southeastern U. S. 1304. 1903, and ed. 2, 1913; Greenm. in Gray, Manual, ed. 7, 854. 1908. S. Balsamitae Ell. Sketch 2:330. 1824, not Muhl. S. aureus var. Balsamitae Gray, Syn. Fl. N. Am. 12:391. 1884, and ed. 2. 1886, in part; Chapman, Fl. Southern U. 8. 245. 1860, ed. 2, 1889, and ed. 3, 266. 1897. S. aureus var. angustifolius Britt. Mem. Torr. Bot. Club 2:39. 1890, not S. angustifolius Willd. S. Earlei Small, Bull. Torr. Bot. Club 25:147. 1898. (?) S. Memmingeri Britt. Bull. Torr. Bot. Club 25:147. 1898. An herbaceous perennial; stems one to several, erect from a соттоп base, 2 to 6 dm. high, conspicuously and perma- nently woolly tomentose at the base and in the leaf-axils ; lower leaves long-petiolate, narrowly oblong-oblanceolate, 1.5 to 13 em. long, 1 to 3 em. broad, obtuse to rounded at the apex, erenate to serrate-dentate, narrowed at the base into the petiole; petioles 1.5 to 18 em. long; stem-leaves petiolate and sublyrate to sessile and more or less pinnatisect; inflo- rescence a terminal many-headed corymbose суше; heads usually very numerous, relatively small, 7 to 10 mm. high, radiate; involuere eampanulate, sparingly calyculate; bracts of the involucre 13 to 21, linear-lanceolate, 5 to 6 mm. long, glabrous; ray-flowers 8 to 13, rays yellow; disk-flowers num- erous; achenes usually but not always hispidulous along the angles. Distribution: southern Pennsylvania to Florida. [VoL. 3 156 ANNALS OF THE MISSOURI BOTANICAL GARDEN Specimens examined: Pennsylvania: Tullytown, Bucks Co., 15 May, 1896, Craw- ford (Phil. Acad. Nat. Sei. Herb.) ; Westtown, Chester Co., 22 June, 1895, Crawford (Phil. Acad. Nat. Sei. Herb.) ; near Mt. Hope, іп red sandstone, Lancaster Co., 24 June, 1901, Heller (Mo. Bot. Gard. Herb.). Maryland: Baldfriar, Cecil Co., 4 July, 1907, Bartram (Phil. Acad. Nat. Sci. Herb.) ; Norbeck, 25 June, 1895, Mearns (U. S. Nat. Herb.); between Garrett Park and Kensington, 8 June, 1907, Steele (U. S. Nat. Herb.) ; field above Cabin John, 7 June, 1908, Steele (U. S. Nat. Herb.) ; dry slopes, Chevy Chase Lake, 30 May, 1911, Standley 5983 (U. S. Nat. Herb.). Distriet of Columbia: vieinity of Washington, 10 June, 1877, and 27 May, 1878, Ward (U. S. Nat. Herb. 131112 in part, and 131111) ; Queen's Chapel Road, 12 June, 1888, Burgess (U. 8. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Garrett Park, June, 1894, and in open thickets, Brookland, 6 June, 1895, Holm (Mo. Bot. Gard. Herb.); Rock Creek Park, 26 May, 1895, Pollard 285 (U. S. Nat. Herb.); vicinity of Washington, 25 May, 1896, Steele (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; wooded side-hill of Piney Branch, Rock Creek Park, 27 Мау, 1899, Maxon 140 (U. 5. Nat. Herb.); Chevy Chase, 16 May, 1905, House 721 (Mo. Bot. Gard. Herb.); Tenallytown, 14 June, 1871, F. Blanchard (Mo. Bot. Gard. Herb.). Virginia: Fairfax Co., 7 June, 1902, G. S. Miller (U. S. Nat. Herb.) ; West End, Fairfax Co., 30 June, 1907, Steele (U. S. Nat. Herb.) ; dry woods, Falls Church, 30 May, 1912, Ruth 164 (U. S. Nat. Herb.) ; open woods, Bluemont, Loudon Co., 31 May, 1915, Standley 11621 (U. S. Nat. Herb.); dry fields, Hampton, 19 May, 1903, G. S. Miller (U. S. Nat. Herb.) ; Rich- mond, de Chalmont (U. S. Nat. Herb.) ; near Suffolk, Nanse- mond Co., 19 May, 1898, Kearney 1271 (U. 5. Nat. Herb.) ; near Branchville, Southampton Co., 12 June, 1893, Heller 958 (Phil. Aead. Nat. Sei. Herb. and Mo. Bot. Gard. Herb.) ; in old fields, vicinity of Belfield, 21 May, 1904, Meyncke (U. S. Nat. Herb.); Appomattox, Rumer (U. S. Nat. Herb.) ; pine 1916] GREENMAN—MONOGRAPH OF SENECIO 157 woods, Bedford Co., 4. H. Curtiss (Gray Herb. and Mo. Bot. Gard. Herb.); Peaks of Otter, 6 June, 1890, Brown, Hogg, Vail, Timmerman & Britton (U. S. Nat. Herb. and Greene Herb.) ; between Fall Creek and Danville, 3 June, 1891, Small £ Heller 234 (U. S. Nat. Herb.) ; vicinity of Marion, Smyth Co., alt. 640 m., 6 June, 1892, N. L. and E. Britton Ф Vail (Phil. Acad. Nat. Sci. Herb.); Hutton's Branch, East Marion, alt. 760 m., 6 June, 1892, Small (Mo. Bot. Gard. Herb.) ; along Comer Creek, 15 June, 1892, alt. 915 m., Small (U. S. Nat. Herb.). North Carolina: Weldon, Williamson (C. S. Williamson Herb.) ; vicinity of Heilig's Mill, Rowan Co., 4-9 June, 1891, Small £ Heller 490 (Mo. Bot. Gard. Herb.) ; in woods, near Faith, 27 May, 1891, Heller 10263 (Mo. Bot. Gard. Herb.) ; Statesville, Hyams (Mo. Bot. Gard. Herb.); near Hickory, alt. 550 m., 23 June, 1893, Heller (Phil. Acad. Nat. Sci. Herb.) ; east of Blowing Rock, Caldwell Co., 24 June, 1893, alt. 1065 m., Heller (Phil. Acad. Nat. Sci. Herb.) ; dry open woods and clearings, Biltmore, 14 June, 1900, Mohr (U. S. Nat. Herb.) ; vieinity of Asheville, May, 1888, McCarthy (U. S. Nat. Herb.); dry woods and waste places, Biltmore, 20 May, 1896, Biltmore Herb. 1233 (Mo. Bot. Gard. Herb.) ; old fields, Bilt- more, 11 June, 1897, Biltmore Herb. 1233b (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; “Half Way," Black Mountain, 27 June, 1902, Harshberger 79 (U. S. Nat. Herb. and Phil. Aead. Nat. Sci. Herb.); Tryon, Polk Co., 22 May, 1899, Churchill (Gray Herb.); foot of mountain, Tryon, 22 May, 1897, Townsend (U. S. Nat. Herb.); Sunburst, Hay- wood Co., alt. 975 m., 19 June, 1910, House 4339 (U. S. Nat. Herb.); without definite locality, June, 1879, Gray, Sargent, Redfield & Canby (Gray Herb.) ; on mountain bluff, Gray Ф Carey (Gray Herb.), small form. South Carolina: Table Rock, Pickens Co., Buckley (Gray Herb.) ; dry woods and fields, Fort Hill, Oconee Co., 30 April, 1906, House 1994 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; without locality, May, 1867, Ravanel (Gray Herb.). Georgia: Gainesville, 10 May, 1879, T'rrelease (Mo. Bot. Gard. Herb.) ; dry woods on highest summit of Pigeon Moun- [Vor. 3 158 ANNALS OF THE MISSOURI BOTANICAL GARDEN tains, Walker Co., alt. 710 m., 1 Aug., 1900, Harper 338 (U.S. Nat. Herb.) ; Lookout Mountain, July, 1898, Ruth 673 (U.S. Nat. Herb.) and 704 (Mo. Bot. Gard. Herb.) ; Lookout Moun- tain, 25 May, 1901, Trelease (Mo. Bot. Gard. Herb.) ; Thomp- son's Mills and vicinity, Gwinnett Co., 26 April, 1908, Allard 267, 268 (U. S. Nat. Herb.) ; Stone Mountain, 13 May, 1901, A. H. Curtiss 6780 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sei. Herb., and Mo. Bot. Gard. Herb.) ; Stone Mountain, 23 May, 1897, and 17 May, 1899, Eggert (Mo. Bot. Gard. Herb.) ; rocky places, Covesprings, 15 May, 1881, Mohr (U.S. Nat. Herb.) ; Tallapoosa, April and May, 1900, Way 30 (U.S. Nat. Herb.) ; Lagrange, 16 May, 1905, Tracy 8944 (U. 5. Nat. Herb. and Mo. Bot. Gard. Herb.) ; without definite locality, Boykin (Phil. Acad. Nat. Sci. Herb.). Alabama: Wedowee, Randolph Co., 28 May, 1874, Mohr (U. S. Nat. Herb.) ; Auburn, Lee Co., colls. of 1897 and 1898, Earle € Baker (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; dry, open places, Auburn, 10 June, 1901, Earle (Gray Herb.) ; Cullman, May, 1886, and June, 1895, Mohr (U. S. Nat. Herb.) ; without definite locality, June, 1889, Miss Emily Mohr (U.S. Nat. Herb. 782485). Florida: without definite locality, Chapman (Gray Herb. and Mo. Bot. Gard. Herb.). Tennessee: dry ground, Knoxville, May, 1898, Ruth 672 (U. S. Nat. Herb.); dry sterile grounds, Knoxville, June, 1898, Ruth 703 (Mo. Bot. Gard. Herb.). This species, S. Small, and the following, S. pauperculus, are, as a rule, easily distinguished by the taller stems, longer leaves, more numerous and somewhat smaller heads of the former; but in southeastern Pennsylvania and in Maryland oceasional forms occur, for example, Crawford's specimen from Tullytown, 15 May, 1896, and Small’s specimen from Mt. Hope, 24 June, 1901, which are somewhat intermediate. These two specimens seem to the writer to possess rather more the characters of S. Small than of S. pauperculus. Moreover, Crawford's specimen from Westtown, Pa., 22 June, 1895, is a perfect match for Heller's specimen, No. 10263, 1916] GREENMAN—MONOGRAPH OF SENECIO 159 from North Carolina, as well as eertain other colleetions from the southern states. The range of S. Smallii may һе said therefore to extend to southern Pennsylvania. 69. S. pauperculus Miehx.' Fl. Bor. Am. 2:120. 1803; Pursh, Fl. Am. Sept. 2:529. 1814, and ed. 2, 1816; DC. Prodr. 6:432. 1837; Britton & Brown, Ill. Fl. 3:545, fig. 4629. 1913. S. Balsamitae Muhl. ex. Willd. Sp. Pl. 3:1998. 1804; Pursh, Fl. Am. Sept. 2:530. 1814, and ed. 2, 1816, excl. synonymy; Beck, Bot. Northern and Middle States 200. 1833, excl. syno- nym; Darlington, Fl. Cest. 497. 1857 ; Heller, Cat. N. Am. PI. 146. 1898, and ed. 2, 229. 1900; Britton & Brown, Ill. Fl. 3:479, fig. 4043. 1898; Britton, Manual 1027. 1901, and ed. 2, 1905, in major part; Greenm. in Rhodora 3:5. 1901; Monogr. Senecio, I. Teil, 23. 1901, and in Engl. Bot. Jahrb. 32:19. 1902; Porter, Fl. Penn. 339. 1903; Keller & Brown, Handb. Fl. Phil. and Vicinity 343. 1905; Greenm. in Gray, Manual, ed. 7, 854. 1908; Small, Fl. Lancaster County 310. 1913. S. aureus var. Balsamitae Torr. & Gray, Fl. N. Am. 2:442. 1843, in part; Torr. Nat. Hist. N. Y., pt. 2, Botany 1:402. 1843; Britton, Cat. Pl. N. J. 54. 1881; ibid. 150. 1890; Gray, Syn. КІ. N. Am. 1?:391. 1884, and ed. 2, 1886, in part; Macoun, Cat. Canadian Pl. 265. 1884, in major part. S. Balsamitae var. praelongus Greenm. Rhodora 3:6. 1901; Graves et al. Conn. Geol. and Nat. Hist. Surv. Bull. No. 14, p. 404. 1910. An herbaceous perennial, somewhat tomentulose, partic- ularly at the base of the stem and in the leaf-axils, to glabrous; stems erect or nearly so, one to several from a common base, 1 to 4 dm. high; lower leaves petiolate, narrowly oblong ob- lanceolate, including the petiole 2 to 18 em. long, .5 to 2 em. broad, rounded to obtuse at the apex, erenate to serrate- dentate, gradually narrowed at the base; stem-leaves petiolate and sublyrate to sessile and pinnatisect; inflorescence a ter- minal few to several-headed corymbose cyme, occasionally re- duced to a single head; heads 5 to 10 mm. high, radiate; in- "The name pauperculus is here maintained for this species instead of Bal- samitae largely on the statement of Dr. Otto Kuntze, Rev. Gen. Pl. 1: CXXXIV, CXXXV. 1891. [Vor. 8 160 ANNALS OF THE MISSOURI BOTANICAL GARDEN voluere campanulate, calyculate; bracts of the involucre (13-) 21, linear-lanceolate, 3 to 6 mm. long, usually glabrous; ray-flowers 8 to 13, rays yellow; disk-flowers numerous; achenes glabrous or hirtellous along the angles. Distribution: Labrador to Minnesota, south to Virginia and Missouri. Specimens examined: Labrador: hills near lighthouse, Forteau, 23 Aug., 1894, Waghorne 29 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; boggy spots, and in wet moss by spring, Blane Sablon, Strait of Belle Isle, 1 Aug., 1910, Fernald £ Wiegand 4185, 4186 (Gray Herb.); wet limestone and ealeareous sandstone ter- races, Blane Sablon, 6 Aug., 1910, Fernald € Wiegand 4178 (Gray Herb.). Newfoundland: damp limestone barrens, near sea-level, Pointe Riche, 4 Aug., 1910, Fernald £ Wiegand 4176 (Gray Herb.) ; dry rocky limestone barrens, near sea-level, Ingorna- choix Bay, 4 Aug., 1910, Fernald € Wiegand 4177 (Gray Herb.) ; barrens at the base of serpentine tablelands, Bonne Bay, 27 Aug., 1910, Fernald & Wiegand 4179 (Gray Herb.) ; crests of sea cliffs, Western Head, New Worlds Island, Notre Dame Bay, 20 July, 1911, Fernald € Wiegand 6407 (Gray Herb.) ; boggy places on hill, southwest of Tilt Cove, Notre Dame Bay, 25 Aug., 1911, Fernald & Wiegand 6412 (Gray Herb.) ; Fogo Island, 27 July, 1903, Sornborger (Gray Herb.) ; ledges and talus, north bank of river below Grand Falls, 3 July, 1911, Fernald € Wiegand 6402 (Gray Herb.) ; open bogs among the hills, Grand Falls, 23 July, 1911, Fernald & Wie- gand 6411 (Gray Herb.) ; bog, Grand Falls, 5 July, 1911, Wil- liamson (C. S. Williamson Herb.) ; shingly beach, north bank of river, below Grand Falls, 22 July, 1911, Fernald € Wie- gand 6406 (Gray Herb.); gravelly river bank, Glenwood, 12 and 13 July, 1911, Fernald & Wiegand 6403, 6404 (Gray Herb.) ; Tilton Harbor to Barred Island, 31 July, 1903, Sorn- borger (Gray Herb.); Barred Island, 13 Aug., 1903, Sorn- borger (Gray Herb.) ; swamp, foot of Helmet, Holyrood, 22 Aug., 1894, Robinson & Schrenk (Gray Herb.) ; near confluence 1916] GREENMAN—MONOGRAPH OF SENECIO 161 of Exploits River and Badger Brook, 13 Aug., 1894, Robinson dé: Schrenk (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; sandy clearing, Mary Anne Brook, 14 July, 1911, Fernald & Wiegand 6410 (Gray Herb.) ; dry bog, Millertown Junction, 7 July, 1911, Fernald & Wiegand 6408 (Gray Herb.) ; damp talus slopes of the marble region between Mt. Musgrave and Humber Mouth (Bay of Islands Station), 18 July, 1910, Fernald & Wiegand 4183 (Gray Herb.); gravelly beach, Middle Birchy Pond, 11 July, 1910, Fernald & Wiegand 4182 (Gray Herb.) ; open tundra along Junction Brook, 12 and 13 July, 1911, Fernald € Wiegand 6409 (Gray Herb.); boggy places on the hill back of Summerside, 11 July, 1910, Fernald & Wiegand 4181 (Gray Herb.) ; open peat bog among Silurian hills, back of Birehy Cove (Curling), 8 July, 1910, Fernald & Wiegand 4180 (Gray Herb.) ; serpentine and magnesian lime- stone barrens, northeastern bases and slopes of Blomidon (Blow-me-down) Mountains, 24 July, 1910, Fernald « Wie- gand 4184 (Gray Herb.); barrens at the base of serpentine table-lands, Bonne Bay, 27 Aug., 1910, Fernald & Wiegand 4187 (Gray Herb.); fields, Coal River, 14 July, 1896, Wag- horne 15 (Gray Herb.), and 16 July, 1896, Waghorne 12 (Mo. Bot. Gard. Herb.) ; sandy shore, Grand Lake, Bay of Islands, 11 Aug., 1896, Waghorne 45 (Gray Herb.); sea-bank, near Chimney Cove, Вау of Islands, Waghorne 8 (Gray Herb.) ; Bay of Islands, 24 June, 1895, Waghorne 23 (Mo. Bot. Gard. Herb.), and 18 July, 1895, Waghorne (U. S. Nat. Herb.) ; open bog, Bay of Islands, 23 July, 1908, Hames & Godfrey 8131 (Gray Herb.). Quebee: rocky shores, River de Brig, Anticosti, 10 July, 1883, Macoun (U. S. Nat. Herb.) ; ravine, Mt. Albert, Gaspé Co., 26 July, 1881, ez Herb. J. A. Allen (U. S. Nat. Herb.) ; crevices and talus of serpentine, gulch north of Lac au Diable, Mt. Albert, Gaspé Co., alt. 750-950 m., 25 July, 1906, Collins & Fernald 752 (Gray Herb.) ; cold wet rocks, head of au Diable, alt. 950 m., 8-15 Aug., 1905, Collins € Fernald (Gray Herb.) ; ealeareous marl, Trout Pond, mouth of Grand River, Gaspé Co. 11-15 Aug., 1904, Collins, Fernald € Pease (Gray 11 [Vor. 3 162 ANNALS OF THE MISSOURI BOTANICAL GARDEN Herb.); banks of Grand River, 20 June-10 July, 1903, Rich- ards (Gray Herb.); banks of Grand River, Gaspé Co., 30 June-3 July, 1904, Fernald (Gray Herb.); gravelly beaches and flats, also wet alluvial shores, between Baldé and Baie des Chaleurs, Bonaventure River, 5, 6 and 7 Aug., 1904, Collins, Fernald € Pease (Gray Herb.) ; Gatineau River, 6 Sept., 1894, Macoun (Mo. Bot. Gard. Herb.); gravel beaches near the mouth of Dartmouth River, 26 and 27 Aug., 1904, Collins, Fernald & Pease (Gray Herb.), form with proliferous heads, also with both radiate and diseoid heads. Ontario: Plevna, 20 June, 1902, Fowler (Gray Herb.) ; sand ridges north of Sarnia, Lambton Co., 13 June, 1895, Dodge 109 (U. S. Nat. Herb.) ; region of Lake Superior, Масош 52 (Mo. Bot. Gard. Herb.). Maine: sunny alluvium, Fort Kent, 6 July, 1904, Fernald (Gray Herb.) ; rocky ledges, Fort Kent, 19 July, 1908, Mac- kenzie 3600 (Mo. Bot. Gard. Herb.); rocky river-flat, Fort Kent, 10 July, 1908, Mackenzie 3418 (Mo. Bot. Gard. Herb.) ; gravelly shores, Fort Fairfield, 5 July, 1893, Fernald 71 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; Pame- dumeook Lake, 10 Aug., 1881, Chickering (Gray Herb. and U. S. Nat. Herb.) ; rocks in river, Orono, 1 July, 1890, Fernald (Gray Herb.). New Hampshire: Sumner’s Falls, 27 June, 1898, Eggles- ton (Mo. Bot. Gard. Herb.), and 25 June, 1902, Eggleston 2804 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.). Vermont: Colchester, coll. of 1842, Oakes (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sei. Herb., and Mo. Bot. Gard. Herb.) ; Winooski High Bridge, 14 June, 1881, Е. Ф C. E. Faxon (Gray Herb.) ; Barnet, 20 June, 1884, F. Blanch- ard (Mo. Bot. Gard. Herb.); Manchester, 27 June, 1898, Day 210 (Gray Herb. and U. S. Nat. Herb.) ; Bellows Falls, coll. of 1901, W. H. Blanchard (Gray Herb.) ; Putney, 8 June, 1902, W. H. Blanchard (Gray Herb.); without definite lo- eality, Robbins (Phil. Acad. Nat. Sei. Herb.). Massachusetts: Danvers, without date, Oakes (Phil. Acad. Nat. Sci. Herb.); without definite locality, Oakes (Gray 1916] GREENMAN—MONOGRAPH OF SENECIO 168 Herb.) ; rocky woods near summit of Blue Hill, Milton, Rich (Gray Herb.). New York: Watertown, Jefferson Co., coll. of 1834, Torr. £ Gray, Fl. N. Am. (Gray Herb.) ; rocky banks of Blue River, Watertown, 3 July, 1857, ex Herb. Wm. Boott (Gray Herb.) ; Dexter, ex Herb. Geo. Vasey (Gray Herb.) ; Oneida Co., coll. of 1864, Paine (Gray Herb.); sandy bog, near Syracuse, 22 June, 1882, Sheldon (Phil. Acad. Nat. Sei. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.); tamaraek swamp, near Syraeuse, 28 May, 1902, House (U. S. Nat. Herb.). New Jersey: low meadows, Newfoundland, Morris Co., 14 June, 1908, Mackenzie 3119 (U. S. Nat. Herb. and Mo. Dot. Gard. Herb.); dry woods, Chatham, 30 May, 1903, Macken- eie 182 (Mo. Bot. Gard. Herb.) ; dry fields, Murray Hill, Union Co., 30 May, 1906, Mackenzie 2044 (Mo. Bot. Gard. Herb.) ; Somerset Co., Perry (Mo. Bot. Gard. Herb.). Pennsylvania: Laanna, Pike Co., 9 June, 1906, Long (Phil. Acad. Nat. Sei. Herb.) ; Allentown, 18 June, 1899, Dowell (U. S. Nat. Herb.); Manganese Park, southeast of Allentown, 8 June, 1908, near Mountainville, 13 June, 1908, and 13 June, 1914, also near Lanark, 24 May, 1908, Pretz 1232, 1253, 6636, 1197 (Phil. Acad. Nat. Sei. Herb.) ; Nockamixon, Bucks Co., June, 1892, Crawford (Phil. Acad. Nat. Sci. Herb.) ; Rockhill, 19 June, 1892, MacElwee (Phil. Acad. Nat. Sci. Herb.) ; Perk- asie, June, 1881, Fretz (Phil. Acad. Nat. Sci. Herb.) ; Tunnel Hill near Perkasie, 31 May, 1903, MacElwee (Phil. Acad. Nat. Sei. Herb.) ; Perkasie, 30 May, 1906, Brown (Phil. Acad. Nat. Sci. Herb.) ; near Sellersville, 7 June, 1893, Porter (Phil. Acad. Nat. Sei. Herb.) ; along Perkiomen Creek, near Sellersville, 2 June, 1899, MacElwee 408 (Phil. Acad. Nat. Sei. Herb.) ; Ar- gus, 6 June, 1898, Fretz (Phil. Acad. Nat. Sci. Herb.) ; Aque- tong, 30 May, 1910, Keller (Phil. Acad. Nat. Sei Herb.): Sumneytown, 30 May, 1903, Jahn, and 30 May, 1905, K elle: (Phil. Aead. Nat. Sci. Herb.); Sumneytown, 50 May, 1903, Williamson (C. S. Williamson Herb.) ; dry fields along Wissa- hickon Creek, Penllyn, 12 June, 1909, Long (Phil. Acad. Nat. Sci. Herb.) ; between Hillside and Ardsley, 12 Oct., 1907, Long [Vor. 3 164 ANNALS OF THE MISSOURI BOTANICAL GARDEN (Phil. Acad. Nat. Sci. Herb.); near Noble, 13 June, 1912, Long 7085 (Phil. Acad. Nat. Sci. Herb.) ; Layfayette, 2 June, 1895, Keller, Jahn, also Uselma C. Smith 816 (Phil. Acad. Nat. Sci. Herb.) ; Layfayette, 5 June, 1897, Jahn (Phil. Acad. Nat. Sei. Herb.) ; serpentine barrens, Newtown, Delaware Co., 25 June, 1901, Benj. H. Smith (Phil. Acad. Nat. Sei. Herb.) ; ser- pentine barrens, near Newtown Square, 12 June, 1899, Mac- Elwee 491, 505 (Phil. Acad. Nat. Sei. Herb.) ; Williamson, 4 June, 1891, Crawford, 28 July, 1899, MacElwee 999, and 11 June, 1911, Pennell 2764, 3641 (Phil. Acad. Nat. Sci. Herb.) ; Media, 30 May, 1896, Githens (Phil. Acad. Nat. Sci. Herb.) ; Elwyn, 8 June, 1890, Brinton, and 9 June, 1890, ex Herb. Ber- inger (Phil. Acad. Nat. Sci. Herb.) ; Wawa, 5 July, 1908, Pen- nell 15, and 9 June, 1896, Bartram (Phil. Acad. Nat. Sei. Herb.) ; Chester Heights, 23 June, 1907, Pennell (Phil. Acad. Nat. Sei. Herb.) ; Mineral Hill, 9 June, 1911, and 6 Sept., 1908, Pennell 2719, 610 (Phil. Acad. Nat. Sci. Herb.) ; Fawkes Run, 30 May, 1909, and 10 June, 1911, Pennell 1365, 2742 (Phil. Acad. Nat. Sci. Herb.) ; Phoenixville, Chester Co., coll. of 1865, Martindale (Phil. Acad. Nat. Sci. Herb.) ; Sugartown Barrens, 22 July, 1908, Pennell 293, and serpentine ridge, northwest of Sugartown, 30 May, 1909, Pennell 1379 (Phil. Acad. Nat. Sci. Herb.); serpentine ridge, Willistown, 28 May, 1904, Painter 673 (U.S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; Wil- listown barrens, 24 May, 1908, Williamson (Phil. Acad. Nat. Sci. Herb.) ; serpentine ridge, Chester Co., June, 1882, Windle (U.S. Nat. Herb.) ; Westtown, 22 June, 1895, Crawford (Phil. Acad. Nat. Sci. Herb.) ; Fern Hill, 7 June, 1909, Long (Phil. Acad. Nat. Sci. Herb.) ; West Chester, coll. of 1828, Wm. Darlington (Phil. Acad. Nat. Sci. Herb.); West Chester, without date, David Townsend, and 29 May, 1909, Pennell 1344 (Phil. Acad. Nat. Sci. Herb.) ; Seonnelltown, 29 July, 1908, Pennell 343 (Phil. Acad. Nat. Sci. Herb.) ; Nottingham Bar- rens, 22 May, 1912, Pennell & Long 8715, 7538 (Phil. Acad. Nat. Sei. Herb.) ; Cedar Barrens, Chester Co., 30 May, 1909, and 10 June, 1911, Pennell 1395, 2760 (Phil. Acad. Nat. Sci. Herb.) ; high above Muddy Run, 5 July, 1904, Crawford (Phil. 1916] GREENMAN—MONOGRAPH OF SENECIO 165 Acad. Nat. Sci. Herb.) ; Pequea, June, 1893, Eby (Mo. Bot. Gard. Herb.); near Mt. Hope, 5 June, 1900, Heller (Gray Herb. and U. S. Nat. Herb.) ; Conewago, 28 May, 1889, Heller (Gray Herb.) ; in wet meadow, Rock Hill, 31 May, 1903, Mac- Elwee (Phil. Acad. Nat. Sci. Herb.) ; Rawlinsville, coll. of 1884, ex Herb. Galen (Gray Herb.) ; Fulton, June, 1906, Car- ter (Phil. Acad. Nat. Sci. Herb.); near Pleasant Grove, in serpentine barrens, 5 June, 1901, Heller (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); dry ground, near Round Top, Gettysburg, Adams Co., 24 June, 1894, MacElwee (Phil. Acad. Nat. Sci. Herb.); York Co., 3 June, 1895, Glat- felter (Mo. Bot. Gard. Herb.); Ohiopyle, Fayette Co., 3-8 June, 1905, Brown, Crawford & Van Pelt (Phil. Acad. Nat. Sci. Herb.). Delaware: dry serpentine barrens, near Centerville, 15 June, 1866, and 10 July, 1868, Commons (Phil. Acad. Nat. Sci. Herb.) ; on serpentine, south of Mount Cuba, 18 June, 1914, Pennell 1510 (Phil. Acad. Nat. Sci. Herb.) ; near Cooch’s Mill, 29 May, 1896, Commons (Phil. Acad. Nat. Sci. Herb. 543180), in part. Maryland: Rockville, 20 May, 1905, Painter 1366 (Mo. Bot. Gard. Herb.) ; Brooklyn, 30 May, 1899, Thurston (U. S. Nat. Herb.) ; Laurel, 23 Мау, 1897, Knowlton (U. S. Nat. Herb.) ; College Park, 28 May, 1900, Stewart (Mo. Bot. Gard. Herb.) ; Mountain Lake Park, Garrett Co., Shreve 560 (U. S. Nat. Herb.). Virginia: on rocks in moss, Great Falls, June, 1903, Painter 357 (U. S. Nat. Herb.). Michigan: “L. Superior, Pie to Sault", Loring (Gray Herb.) ; Macinaw City, 12 Aug., 1890, Wheeler (Gray Herb.) ; Thunder Bay Island, Alpena Co., 18 July, 1895, Wheeler (Gray Herb.) ; North Point, Alpena Co., 3 July, 1895, Wheeler (U. S. Nat. Herb.); dry woods near Hillman, Montmorency Co., 7 July, 1895, Wheeler (U. S. Nat. Herb.) ; Clarkston, 30 June, 1888, Hicks (U. S. Nat. Herb.) ; marl beds on Parkdale Farm, colls. of 1912, 1913, and 1914, Farwell 2652, 2653, 3491, 3659, 3660 (Mo. Bot. Gard. Herb.). [VoL. 3 166 ANNALS OF THE MISSOURI BOTANICAL GARDEN Ohio: Marblehead Peninsula, 20 May, 1895, Moseley (Г. S. Nat. Herb.). Indiana: Mineral Springs, 14 June, 1911, Nieuwland 2657 (Mo. Bot. Gard. Herb.); moist sands and swales, Pine, 12 June, 1897, Umbach (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; sandy soil, Edgmoor, Lake Co., 13 June, 1891, Mof- fatt 451 (Field Mus. Herb.) ; prairie, Roby, 18 June, 1910, Lan- sing 2784 (U.S. Nat. Herb.). Minnesota: Center City, June, 1892, Taylor (U. S. Nat. Herb.); damp places near Minneapolis, June, 1892, Burgle- haus (Mo. Bot. Gard. Herb.) ; without definite locality, July, 1899, Woods (U. S. Nat. Herb.) ; without locality, June, 1849, Dr. Sykes (Mo. Bot. Gard. Herb.). Wisconsin: “N. W. America", Long (Mo. Bot. Gard. Herb.). Illinois: in moist sandy soil near Lake Michigan, Beach, 16 June, 1907, Greenman 1987, 2015 (Mo. Bot. Gard. Herb.); rich meadow, Beverly Hills, 6 June, 1903, Chase 2066 (U. S. Nat. Herb.) ; Morgan Park, 27 May, 1907, Dixon & Gage 710 (U. S. Nat. Herb.). Missouri: near Mark Twain's Cave, Ralls Co., 5 May, 1914, Rev. John Davis 2333 (Mo. Bot. Gard. Herb.). Var. firmifolius (Greenm.) comb. nov. S. Balsamitae var. firmifolius Greenm. Rhodora 7:244. 1905. Stems .5 to 2.5 dm. high, simple or branched, more or less tufted; lower leaves mostly short-petiolate, subrotund, ob- long-elliptie to oblong-oblanceolate, .5 to 4 em. long, 5 to 20 mm. broad, crenate-dentate to sublyrately pinnatifid, at first as well as the stem somewhat tomentulose, later more or less glabrate and thickish or firm in texture; upper stem-leaves sessile and pinnatisect to linear and bracteiform. Distribution: in limestone detritus, crest of Les Murailles, Percé, Gaspé Co., 17 Aug., 1904, Collins, Fernald & Pease (Gray Herb.), түре; limestone detritus, Mont Rouge, Percé, Gaspé Co., 23 July, 1905, Collins & Fernald (Gray Herb.) ; limestone detritus, Cap Barré, Pereé, Gaspé Co., 23 July, 1916] GREENMAN—MONOGRAPH OF SENECIO 167 1905, Collins & Fernald 147 (Gray Herb. and U. S. Nat. Herb.) ; limestone shingle near summit of Baldé, Bonaventure Co., 5, 6, and 8 Aug., 1904, Collins, Fernald & Pease (Gray Herb.) ; wet red sandstone bluffs and steep slopes, between Baldé and the Baie des Chaleurs, Bonaventure River, 5, 6, and 8 Aug., 1904, Collins, Fernald & Pease (Gray Herb.) ; ledgy banks of the Restigouche River, Metapedia, 19 July, 1904, Collins & Fernald (Gray Herb.). The last three speci- mens cited are transitional forms between the variety and the species. 70. S.flavovirens Rydb. Bull. Torr. Bot. Club 27:181, pl. 5, fig. 4. 1900, mainly ; Greenm. Monogr. Senecio, I. Teil, 24, 1901, and in Engl. Bot. Jahrb. 32:20. 1909. S. fulgens Rydb. Bull. Torr. Bot. Club 27:177, pl. 6, fig. 13. 1900, not. Nichols. S. Rydbergu Nels. in Coulter & Nelson, Manual Cent. Rocky Mountains 582. 1909. S. Balsamitae Rydb. Mem. N. Y. Bot. Gard. 1:446. 1900, mainly, not Muhl.; Nelson in Coulter & Nelson, Manual Cent. Rocky Mountains 583. 1909, not Muhl. S. flavulus Rydb. Fl. Colo. 397. 1906, in part, not Greene. An herbaceous perennial, pale or yellowish green in the dried state, glabrous or slightly white-tomentulose along the margins of the petioles near their base and in the axils of the leaves; stems erect, 2 to 5 dm. high; lower leaves petiolate, oblanceolate to oval, 1 to 6 em. long, .5 to 2 em. broad, grad- ually narrowed into the petiole to abruptly constricted at the base, crenate to coarsely and unequally dentate, obtuse or rounded at the apex, glabrous on both surfaces; stem-leaves petiolate and more or less lyrate to sessile and pinnatisect; inflorescence a terminal few-headed corymbose cyme; heads Т to 9 mm. high, radiate; involucre campanulate, calyculate; bracts of the involucre 13 to 21, linear-lanceolate, 5 to 7 mm. long, pale or yellowish green and glabrous except at the brownish penicillate tip, becoming thickish in texture; ray- flowers 10 to 12, rays bright yellow; disk-flowers numerous; achenes glabrous or slightly hirtellous along the angles. [Vor. 3 168 ANNALS OF THE MISSOURI BOTANICAL GARDEN Distribution: British Columbia, south to Colorado and Idaho. Specimens examined: British Columbia: Field, 12 July, 1904, Farr (Univ. Penn. Herb.) ; Ottertail Drive, near Field, 15 July, 1905, Farr 817, 818 (Univ. Penn. Herb.) ; border of an alkali marsh, Similka- meen River, 10 June, 1905, Macoun 69356 (Gray Herb.); Lower Fraser River, N. Lat. 49°, Oregon Boundary Commis- sion, coll. of 1859, Dr. Lyall (Gray Herb.). Montana: Big Fork, Aug., 1908, Mrs. Joseph Clemens (Field Mus. Herb. and Mo. Bot. Gard. Herb.). Yellowstone National Park: Mammoth Hot Springs, 5 July, 1899, Blankenship (Gray Herb.) ; about willow elumps on river bottom, Snake River, 12 Aug., 1899, 4. «€ E. Nelson 6402 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); Fitz- gerald's Ranch, near Gardiner Mountain, 3 July, 1902, Mearns 1493 (U. S. Nat. Herb. and Field Mus. Herb.). Wyoming: low ground, Adam's Ranch, Jackson's Hole, 15 July, 1901, Merrill & Wilcox 967 (U. S. Nat. Herb.) ; sandy soil, Blue Lakes on Wind River, 6 July, 1881-82, Forwood (U. S. Nat. Herb.); Wind River, Aug., 1894, A. Nelson 760 (Gray Herb. and U. S. Nat. Herb.); Horse Creek, coll. of 1893, A. Nelson 100 (U. S. Nat. Herb.) ; Green River, 26 July, 1894, A. Nelson 1036 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; Dale Creek, near Sherman, 29 July, 1884, Letterman (Mo. Bot. Gard. Herb.) ; Evanston, 10 July, 1897, Williams (U. S. Nat. Herb.) ; La Barge, Uinta Co., 18 July, 1894, Stevenson 201 (U. S. Nat. Herb.) ; Myer's ranch, Bear River, south of Evanston, alt. 2285—2440 m., 26 July, 1902, Pammel 6 Blackwood 4044 (Mo. Bot. Gard. Herb.). Colorado: N. Lat. 39-412, coll. of 1862, Hall £ Harbour 332, in part (Phil. Aead. Nat. Sei. Herb. and Mo. Bot. Gard. Herb.), and 115, in part (Gray Herb.); Colorado Springs, Мау, 1878, Jones (U. б. Nat. Herb. 223024, in part). Idaho: open grassy stream lands, Mackay, Custer Co., 1 Aug., 1911, Nelson & Macbride 1500 (Mo. Bot. Gard. Herb.) ; Areo, 19 June, 1893, E. Palmer 193 (U. S. Nat. Herb.). 1916] GREENMAN—MONOGRAPH OF SENECIO 169 Var. thomsoniensis (Greenm.) comb. nov. S. Balsamitae var. thomsoniensis Greenm. Ottawa Nat. 25 :116. 1911. S. Balsamitae Greenm. Ottawa Nat. 25:116. 1911, not Muhl. Stems 1.2 to 4.5 dm. high, at first floccose-tomentulose later more or less glabrate; lower leaves oblong-oblanceolate, the blade 1 to 7 em. long, 5 to 12 mm. broad, rounded or obtuse at the apex, crenate to serrate, gradually narrowed at the base into the petiole, at first tomentulose, particularly on the under surface, later more or less glabrate; inflorescence and base of the involucre often slightly tomentulose. Distribution: Alaska to British Columbia and Montana. Specimens examined: Alaska: on gravel flood-plain of the Kuskokwim River, 19 July, 1902, Brooks Ф Prindle (U. S. Nat. Herb.). British Columbia: Bonaparte River, 18 June, 1889, Macoun (U. S. Nat. Herb. 219791, in part) ; South Thompson River at Kamloops, 10 July, 1906, E. Wilson 686, 672 (Mo. Bot. Gard. Herb. and Geol. Surv. Canada Herb. 81261) ; Lake Osoyoos, 31 May, 1905, Macoun 69357 (Gray Herb.). Montana: Big Fork, Flathead Co., 14 June, 1904, W. W. Jones (Mo. Bot. Gard. Herb.). Washington: Fort Okanogan, U. S. Exploring Expedition, Wilkes 971 (С. S. Nat. Herb. 48747). 71. S. multnomensis Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Ottawa Nat. 25:115. 1911. An herbaceous perennial, glabrous or slightly floecose- tomentulose in the axils of the leaves, on the margins of the petioles, and in the inflorescence; stems solitary or cespitose, erect or somewhat flexuous, 3 to 7 dm. high; lower leaves ob- long-oblaneeolate including the petiole 4 to 15 em. long, .8 to 2 em. broad, obtuse or rounded at the apex, erenate-dentate to more or less lyrately lobed with remote lobes and deep rounded sinuses, narrowed at the base into a slender petiole usually exceeding the blade; upper stem-leaves sessile and pinnatisect to much reduced entire bracts; inflorescence a ter- [Vor. 3 170 ANNALS OF THE MISSOURI BOTANICAL GARDEN minal few to many-headed corymbose cyme; heads 10 to 13 mm. high, radiate; involucre campanulate, sparingly calyecu- late; bracts of the involuere about 21, linear-lanceolate, 7 to 10 mm. long, usually pale green, glabrous or slightly floceose- tomentulose; ray-flowers 10 to 13, rays yellow; disk-flowers numerous, about 60; achenes glabrous. Distribution: Mackenzie south to Saskatchewan, west to eastern Washington. Specimens examined: Mackenzie: Fort Smith, 4 Aug., 1901, Е. А, & A. E. Preble 172 (U. B. Nat. Herb.). Saskatchewan: without definite locality, Palliser’s British N. Am. Expl. Expedition, 1858, E. Bourgeau (Gray Herb.) ; Cypress Hills, province of Assiniboia, 24 June, 1894, Macoun 5070 (Gray Herb. and U. 8. Nat. Herb.). Alberta: dry or rocky soil, Bow Valley, five miles west of Calgary, 14 June, 1913, Moodie (U.S. Nat. Herb.). British Columbia: Kicking Horse Valley, vicinity of Field, alt. 1220 m., 20 June-25 July, 1906, Brown 486, 487 (Phil. Acad. Nat. Sci. Herb.); Glacier, 30 July, 1901, Williamson (С. S. Williamson Herb.) ; cleared land at Howser Lake, alt. 610 m., 15 June, 1905, Shaw 722 (U. S. Nat. Herb.) ; Trail, 10 June, 1902, Macoun 64989 (Gray Herb.) ; Trail, 18 June, 1902, Macoun 64992 (Gray Herb. and Mo. Bot. Gard. Herb.) ; rocky valley of Fraser River, above Yale, 22 July, 1880, G. Engel- mann (Mo. Bot. Gard. Herb.). Idaho: wet soil, Kootenai Co., alt. 900 m., July, 1900, Lei- berg (Mo. Bot. Gard. Herb.). Washington: Coleville Reservation, June, 1902, Griffiths Ф Cotton 366 (U. S. Nat. Herb.). Oregon: sandy flats, Cascades, 25 Мау, 1869, Kellogg d Harford 537 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; Multnomah Co., June, 1877, T. J. Howell 221 (Gray Herb.), түрЕ. 72. 8. laetiflorus Greene, Pittonia 3:88. 1896. S. aureus var. borealis Eaton, in Bot. King's Exp. 190. 1871, not Torr. & Gray. 1916] GREENMAN—MONOGRAPH OF SENECIO 171 S. cymbalarioides var. diversilobus Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. An ровя Ee glabrous, or in the early stages slightly white-fl ] and soon glabrate; stems 2 to 5 dm. high, simple or branched from the base; lower leaves petiolate, broadly ovate, elliptic-oblong to obovate, 1 to 6.5 em. long, .5 to 4.5 em. broad, entire to crenate-dentate, cuneate at the base and more or less decurrent on the petiole, thick and firm in texture; petioles 1.5 to 14 em. long; stem- leaves sublyrate or irregularly pinnate-lobed with remote lobes, the uppermost sessile, semi-amplexicaul, often much reduced; inflorescence a few to many-headed corymbose cyme; heads 8 to 10 mm. high, radiate; involucre campanulate, spar- ingly ealyeulate; bracts of the involucre (13-)21, linear- lanceolate, 5 to 8 mm. long, thickish, glabrous or at first slight- ly tomentulose and glabrate; ray-flowers about 13, rays pale yellow; disk-flowers numerous; achenes glabrous. Distribution: Oregon, California, Idaho, and Nevada. Specimens examined. Oregon: moist meadows, Powder River Valley, alt. 1065 m., June-July, 1897, Cusick 1617 (Gray Herb, U. S. Nat. Herb., Greene Herb., and Mo. Bot. Gard. Herb.) ; Otis Creek, alt. 1100 m., 19 June, 1896, Leiberg 2324 (U. S. Nat. Herb.) ; near Devine ranch, alt. 1290 m., 27 June, 1896, Leiberg 2411 (U. S. Nat. Herb.) ; Harney Valley, 29 May, 1885, Th. Howell (Gray Herb. and U. S. Nat. Herb.) ; near Silver Lake, alt. 1470 m., 20 Aug., 1894, Leiberg 764 (Mo. Bot. Gard. Herb.), and 764a, 7645 (U. S. Nat. Herb.) ; Annie Creek, 8 Aug., 1897, Mrs. R. M. Austin 1618 (U. S. Nat. Herb.). California: near Boca, 26 July, 1895, E. L. Greene (Greene Herb.), түре; Gazelle, Siskiyou Co., 20 June, 1905, Heller 8076 (U. S. Nat. Herb., Phil. Acad. Nat. Sei. Herb., and Mo. Bot. Gard. Herb.) ; Purdy, Sierra Co., 1 July, 1907, Heller ё Kennedy 8665 (U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.); Summit, Bolander € Kellogg (Gray Herb.) ; Sierraville, alt. 1525 m., Най & Babcock 4473 (Gray Herb.) ; Little Truckee River, alt. 2740 m., July, 1903, (eo, 3 172 ANNALS OF THE MISSOURI BOTANICAL GARDEN Hall £ Babcock 4526 (Gray Herb.); Sierra Nevada Moun- tains, coll. of 1875, Lemmon (U.S. Nat. Herb. 48715). Nevada: Hunter Creek Canon, Washoe Co., alt. 1830 m., 18 June, 1912, Heller 10479 (U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.) ; Lemmon Valley, Washoe Co., alt. 1830 m., 8 July, 1913, Kennedy 2066 (Mo. Bot. Gard. Herb.) ; Carson City, alt. 1525 m., 4 June, 1897, M. Е. Jones (U. S. Nat. Herb. 359610) ; near Carson City, coll. of 1865, Anderson (Gray Herb.) ; Ruby Valley, alt. 1830 m., U. S. Geol. Expl. of the 40th Parallel, July, 1868, Watson 670 (Gray Herb. and U. S. Nat. Herb.) ; without definite locality, coll. of 1872, Lieut. Wheeler (U. S. Nat. Herb. 48772). Idaho: in patches on open slopes, Deer Creek, Owyhee Co., 1 July, 1912, Nelson € Macbride 1850 (С. S. Nat. Herb. and Mo. Bot. Gard. Herb.); moist sunny slopes, Three Creek, Owyhee Co., 1 July, 1912, Nelson & Macbride 2247 (0. S. Nat. Herb.). 73. S. Suksdorfii Greenm. Bot. Gaz. 53:511. 1912; Piper & Beattie, Fl. Northwest Coast 389, 1915. S. Adamsi Howell, Fl. Northwest Am. 1:379. 1900, not S. Adamsii Cheesem. Trans. N. Z. Inst. 28:536. 1896; Piper, Contr. U. S. Nat. Herb. 11:598. 1906. An herbaceous perennial, slightly white-floccose-tomentulose in the leaf-axils and on the margins of the petioles near their base, otherwise glabrous; stems one to several from a rather stout rootstock, 1 to 2.5 dm. high; lower leaves petiolate, broadly ovate, 1 to 4 em. long, 1 to 2.5 em. broad, subtruncate to abruptly contracted to a cuneate base, erenate-dentate from base to apex, glabrous on both surfaces; petioles 1 to 5 em. long; stem-leaves petiolate and sublyrate to sessile and more or less incised-dentate; inflorescence a few to several-headed corymbose cyme; heads 8 to 10 mm. high, radiate; ray-flowers 8 to 12, rays yellow ; disk-flowers numerous ; achenes glabrous. Distribution: Washington to Nevada and California. Specimens examined: Washington: Mt. Adams (Paddo), alt. about 2155 m., 9 Aug., 1882, Suksdorf 73 (Gray Herb., Geol Surv. Canada 1916] GREENMAN—MONOGRAPH OF SENECIO ЕГЭ Herb., and Field Mus. Негр.) ; east of Mt. Adams (Paddo), Aug., 1892, Henderson 2309 (Gray Herb.); Mt. Adams, alt. 1980 m., 25 July, 1899, Flett (U. S. Nat. Herb. 415519 in part); Indian Henry Park, Sept., 1909, Tarleton 62 (Field Mus. Herb.). Oregon: bases of granitic cliffs, source of the Imnaha, Wal- lowa Mountains, alt. 2690 m., Aug., 1906, Cusick 3131 (U. S. Nat. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.). Nevada: Mt. Rose, alt. 2940 m., 29 July, 1909, Heller 9896 (U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., Field Mus. Herb., and Mo. Bot. Gard. Herb.). California: wet mountain slopes at Lady Bug Peak, alt. 2440 m., 12 Aug., 1900, Leiberg 5318 (U. S. Nat. Herb.). 74. S. rubricaulis Greene, Pittonia 3:89. 1896. S. aureus var. croceus Eaton in Bot. King's Exp. 190. 1871, not Gray, Proc. Acad. Phil. 15:68. 1863. S. Јопеѕи Rydb. Bull. Torr. Bot. Club 27:179, pl. 5, fig. 5. 1900. S. cymbalarioides Nelson in Coulter E Nelson, Manual Cent. Rocky Mountains 582. 1909, as to S. Jonesii in synonymy. An herbaceous perennial, glabrous or nearly so; stems erect, 3 to 4 dm. high; lower leaves petiolate, broadly ovate to obovate, 1 to 5 em. long, 1 to 3 em. broad, rather coarsely den- tate, abruptly euneate at the base, glabrous on both surfaces; petioles 1 to 7 em. long, often purplish; stem-leaves petiolate and sublyrate to sessile and pinnatifid; inflorescence a ter- minal few to several-headed corymbose eyme; heads 8 to 10 mm. high, radiate; involucre campanulate, sparingly calyeu- late; bracts of the involuere 13 to 21, linear-lanceolate, 5 to 7 mm. long, acute, glabrous, thickish, and in the later stages drying dark brown or blackish; ray-flowers 8 to 10, rays yel- low; disk-flowers numerous; achenes glabrous. Distribution: mountains of Utah, Wyoming, and Nevada. Specimens examined: Nevada: foothills of Clover Mountains, coll. of 1893, E. L. Greene (Greene Herb.), түре; Clover Mountains near Deeth, Elko Co., alt. 2000 m., 22 July, 1908, Heller 9091 (Phil. Acad. Nat. Sci. Herb.). [Vor. 3 174 ANNALS OF THE MISSOURI BOTANICAL GARDEN Utah: Alta Wasatch Mountains, alt. 3350 m., 31 July, 1879, M. E. Jones 1125 (Torrey Herb. and Field Mus. Herb.), TYPE of S. Лопези; Wasatch Mountains, alt. 1525 m., May, 1869, Watson 671 (Gray Herb.) ; Salt Lake City, alt. 1300 m., 15 July, 1880, M. E. Jones 1996 (U.S. Nat. Herb.) ; Red Butte Cañon, Salt Lake Co., 12 July, 1906, Garrett 1854, 1854a (Field Mus. Herb.). Yellowstone National Park: Electric Peak, alt. 2590 m., 26 July, 1902, Sheldon 179 (U. S. Nat. Herb.) ; without definite locality, Aug., 1902, Mearns 2671, 2719 (U. S. Nat. Herb.). Var. aphanactis Greenm. var. nov. Stem somewhat flexuous, at the base as well as the pet- ioles more or less purplish; lower leaves rather coarsely and unequally dentate; heads about 1 em. high, slightly nod- ding, discoid. Distribution: Utah. Specimen examined: Utah: dry сайоп, Logan, Cache Co., alt. 1525 m., 23 June, 1910, C. P. Smith 2208 (Field Mus. Herb., photograph in Mo. Bot. Gard. Herb.), TYPE. 75. S. cymbalarioides Nutt. Trans. Am. Phil. Soc. N. 8. 7:412. 1841; Rydb. Mem. N. Y. Bot. Gard. 1:446. 1900, in ` part; Bull. Torr. Bot. Club 27:178. 1900, mainly; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Piper, Contr. U. S. Nat. Herb. 11:598. 1906. S. aureus var. borealis Torr. & Gray, Fl. N. Am. 2:442. 1843, in part, 1. e., S. cymbalarioides in synonymy; Gray, Syn. Fl. N. Am. 1?:391. 1884, and ed. 2, 1886, in part. S. aureus var. obovatus Eaton, in Bot. King's Exp. 190. 1871, not Torr. & Gray. S. subcuneatus Rydb. Bull. Torr. Bot. Club 27:179, pl. 5. fig. 6. 1900; Fl. Colo. 397. 1906. An herbaceous perennial, glabrous except for a white- Яоссове tomentum in the axils of the leaves and on the base of the petioles; stems solitary or several from a com- mon base, 1 to 3 dm. high; lower leaves petiolate, the blade broadly ovate, obovate to somewhat spatulate, 1 to 6 cm. 1916] GREENMAN—MONOGRAPH OF SENECIO 179 long, .5 to 3 em. broad, entire or dentate towards the apex, glabrous on both surfaces, thiek and firm in texture; pet- ioles 1 to 8 em. long; stem-leaves petiolate or sessile, in- cised-serrate to entire, the uppermost much reduced; in- florescence a few to several-headed corymbose суше; heads 8 to 10 mm. high, radiate; involuere eampanulate, sparingly ealyeulate, glabrous or rarely slightly tomentulose at the base; bracts of the involucre linear-lanceolate, 5 to 8 mm. long, thickish, in age drying dark brown or blackish and appearing somewhat glutinous; ray-flowers 8 to 12, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: Alberta and British Columbia, south to New Mexico and Nevada. Specimens examined. British America: Explorations in subaretie America, coll. of 1862, Onion, Kennicott € Hardisty (U. S. Nat. Herb.). Montana: Duck Lake, 23 June, 1901, Weller (U. S. Nat. Herb.); hills and plains, Midvale, 17 and 24 June, 1903, Umbach 78, 154 (U. S. Nat. Herb.); Little Belt Pass, alt. 2130 m., 10 Aug., 1896, Flodman 910 (U. S. Nat. Herb.) ; near Red Lodge, 28 July, 1893, Rose 79 (U. S. Nat. Herb.) ; Jack Creek Cañon, 15 July, 1897, alt. 2135 m., Rydberg Ф Bessey 5265 (U. S. Nat. Herb.). Yellowstone National Park: Swan Lake Flat, 30 July, 1902, Sheldon 266 (U. S. Nat. Herb.) ; in open aspen groves, Yellowstone River, near Junction Butte, 10 July, 1899, A. Ф E. Nelson 5823 (U. S. Nat. Herb., Gray Herb., and Mo. Bot. Gard. Herb.). Wyoming: mountain sides, head of Middle Fork on Pow- der River, Big Horn Co., 19 July, 1901, Goodding 302 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; grassy hillside, Ten Sleep Lakes, Big Horn Co., 30 July, 1901, Goodding 414 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.); northwestern Wyoming, 4 Sept, 1893, Rose 683 (U. S. Nat. Herb.); flats near Holm Lodge, 28 June, 1911, Reynolds 5 (Field Mus. Herb.); river bottoms, En- [Vor. 3 176 ANNALS OF THE MISSOURI BOTANICAL GARDEN campment, alt. 2195 m., 20 June, 1901, Tweedy 4130 (U. 5. Nat. Herb.). Colorado: Grizzly Creek, 19 July, 1896, alt. 2590 m., Baker (Mo. Bot. Gard. Herb.), со-түре of S. subcuneatus. New Mexico: Baldy, 14 Aug., 1910, Wooton (U. S. Nat. Herb.). Idaho: moranie ridge, south of Petit Lake, alt. 2195- 9985 m., 14 Aug., 1895, Evermann 330 (U. S. Nat. Herb.) ; Beaver Cañon, 28 June, 1895, Shear 3028 (U. S. Nat. Herb.) ; timbered slopes, Mackay (Bear Cafion), Custer Co., alt. 2435 m., 31 July, 1911, Nelson Ф Macbride 1510 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); four miles south of Ketehum, 23 July, 1895, Henderson 3234 (U. S. Nat. Herb.) ; Henry's Lake and Mt. Chauvet, alt. 3050 m., 29 July, 1897, Rydberg & Bessey 5267 (U. S. Nat. Herb. and Kew Herb.). Utah: Wasatch Mountains, alt. 2135 m., June, 1869, Wat- son 669 (Gray Herb.); Fish Lake, alt. 3050 m., 2 Aug., 1894, Jones 57170 (0. S. Nat. Herb. 235767); Thousand Lake Mountain, alt. 3140 m., U. S. Geol. and Geog. Survey of the Territories, 13 July, 1875, Ward 366 (U. S. Nat. Herb. 143111 in part). Nevada: sagebrush flats, Mountain City, alt. 1830 m., 15 Aug., 1912, Nelson Ф Macbride 2201 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); meadow, vicinity of Gold Creek, 6 Aug., 1913, Hitchcock 1054 (U. S. Nat. Herb.). Washington: south slope of Mt. Chapa, alt. 1220 m., Aug., 1897, Elmer 592 (Mo. Bot. Gard. Herb. and Berlin Herb.) ; moist slopes, head of Prince Creek, alt. 1585 m., 2 Sept., 1897, Gorman 809 (U. S. Nat. Herb.); coniferous woods, head of Twenty-five-mile Creek, alt. 1490 m., 10 Aug., 1897, Gorman 810 (U. S. Nat. Herb.) ; slopes of Mt. Stuart, alt. 1065-1830 m., 24 July, 1893, Sandberg & Leiberg 553 (Field Mus. Herb. Greene Herb., and Mo. Bot. Gard. Herb.), co- түре of S. fraternus; Mt. Stuart, Kittitas Co., July, 1898, Elmer 1384 (Mo. Bot. Gard. Herb.) ; Yakima Region, North- ern Transcontinental Survey, coll. of 1883, Brandegee 916 (Gray Herb.). 1916] GREENMAN—MONOGRAPH OF SENECIO 177 Oregon: ‘‘Columbia woods,” Nuttall (Phil. Acad. Nat. Sci. Herb.) ; “В. Mts.,’’ Nuttall (Gray Herb.) ; river bottoms, Union Co., coll. of 1881, Cusick 928 (Gray Herb.) ; Cascade Mountains, alt. 1525 m., Oregon Boundary Commission, coll. of 1860, Lyall (Gray Herb.). Var. borealis (T. & G.) Greenm. comb. nov. S. aureus var. borealis Torr. & Gray, Fl. N. Am. 2:442. 1843, excluding S. cymbalarioides Nutt.; Gray, Syn. Fl. N. Am. 1:391. 1884, and ed. 2, 1886, in small part; Macoun, Cat. Canadian Pl. 265. 1884, in part; Porter & Coulter, Syn. Fl. Colo. 81. 1874, in part; Coulter, Manual Roeky Mountain Region 211. 1885, in part. S. aureus Hook. Fl. Bor. Am. 1:333. 1834, in part, not L. S. elongatus Howell, Fl. Northwest Am. 1:379. 1900, as to S. aureus var. borealis in synonymy, not Pursh. Stems slender, nearly naked above; lower leaves sub- spatulate to oblanceolate, including the petiole 1.5 to 6 em. long, less than 1 em. broad; in all other characters like the species into which it passes through several intermediate forms. Distribution: Arctic America to Wyoming and Utah. Specimens examined: ** Arctic America”: ex Herb. Hooker (Gray Herb.). Alberta: Fort Chipewyan, Athabasca, 5 June, 1903, Preble & Cary 3 (U. S. Nat. Herb.); Peace River Landing, 13 June, 1903, Macoun 61239 (Gray Herb.). British Columbia: Skagit Valley, alt. 1220 m., 10 July, 1905, Масоит 69362 (Gray Herb.). Montana: Monida, 26 June, 1900, Blankenship (Gray Herb.). Wyoming: head of Big Goose Creek, Big Horn Moun- tains, 15-24 July, 1898, Tweedy 65 (U. S. Nat. Herb.). Utah: Thousand Lake Mountain, alt. 3170 m., U. S. Geol. and Geog. Survey of the Territories, 19 July, 1875, Ward 366 (Gray Herb. and U. S. Nat. Herb. 145111 in part). Var. streptanthifolius (Greene) Greenm. comb. nov. 12 [Vor. 3 178 ANNALS OF THE MISSOURI BOTANICAL GARDEN S. streptanthifolius Greene, Erythea 3:23. 1895; Howell, Fl. Northwest Am. 1:375. 1900. Lower leaves suborbicular, obovate or oblong-obovate, somewhat glaucous. Distribution: southeastern Idaho and northwestern Wy- oming. Specimens examined: Idaho: Beaver Cañon, 1 Aug., 1889, E. L. Greene (Greene Herb.), түре; Beaver Cañon, 29 July, 1889, E. L. Greene (U. S. Nat. Herb.) ; northwestern Wyoming, 22 Aug., 1893, Rose 243 (U. S. Nat. Herb.). 76. S. acutidens Rydb. Bull. Torr. Bot. Club 27:180, pl. 5, fig. 2. 1900; Greenm. Monogr. Senecio, I. Teil, 23. 1901, and in Engl. Bot. Jahrb. 32:19. 1902. S. cymbalarioides Coulter & Nelson, Manual Cent. Rocky Mountains 582. 1909, in part, as to S. acutidens in synonymy; Wooton & Standley, Contr. U. S. Nat. Herb. 19:747. 1915, as to specimen cited. An herbaceous perennial, glabrous or slightly floccose- tomentulose in the early stages and soon glabrate except in the axils of the leaves; stems 1.5 to 2.5 dm. high, erect, more or less tufted, simple or branched; lower leaves obovate to oblong-oblanceolate, including the petiole 3 to 12 em. long, .9 to 1.5 em. broad, dentate towards the apex, entire towards the base and gradually narrowed into the petiole, thick and firm in texture, often bluish green and somewhat glaucous; stem-leaves oblanceolate and rather sparingly dentate to lance-attenuate and entire; inflorescence a terminal few to several-headed corymbose суше; heads 8 to 10 mm. high, radiate; involuere sparingly calyculate; ray-flowers 8 to 10, rays yellow; disk-flowers numerous; achenes glabrous or rarely slightly hirtellous along the angles. Distribution: Wyoming to New Mexico. Specimens examined: Wyoming: Union Pass, Wind River Range, 10 Aug., 1894, 4. Nelson 858 (N. Y. Bot. Gard. Herb., Gray Herb., U. 8. 1916] GREENMAN—MONOGRAPH OF SENECIO 179 Nat. Herb., Greene Herb., and Mo. Bot. Gard Herb.), түрЕ; near Fort Bridger, coll. of 1873, Pruddon (Gray Herb.). Colorado: Denver, Lieut. Wheeler’s Expedition, 1873, Wolf £ Rothrock 558 (Gray Herb. and С. S. Nat. Herb.) ; Union Creek Pass, Lieut. Wheeler’s Expedition, 1873, Wolf & Rothrock 586 (U. S. Nat. Herb. and Gray Herb.). New Mexico: Costilla Valley, alt. 3000 m., 5 Sept., 1913, Wooton (U. S. Nat. Herb.). 77. S.tridenticulatus Rydb. Bull. Torr. Bot. Club 27:175. 1900; Fl. Colo. 396. 1906. S. compactus (Gray) Rydb. Mem. Torr. Bot. Club 5:342. 1894, not Kirk in Trans. N. Z. Inst. 12:395. 1880; Heller, Cat. N. Am. Pl. 146. 1898, and ed. 2, 229. 1900; Britton & Brown, Il. Fl. 3:480, fig. 4045. 1898; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Britton, Manual 1028. 1901, and ed. 2, 1905. S. aureus var. compactus Gray, Syn. Fl. N. Am. 1?:391. 1884, and ed. 2, 1886. S. aureus var. borealis Gray, Pl. Wright., pt. 1, p.125. 1852, not Torr. & Gray. S. oblanceolatus Rydb. Bull. Torr. Bot. Club 27:175, pl. 5, fig. 9. 1900; Fl. Colo. 396. 1906; Wooton & Standley, Contr. U. S. Nat. Herb. 19:747. 1915. S. densus Greene, Pittonia 4:226. 1900; Britton & Brown, Ш. Fl. 3:546, fig. 4631. 1913. S. condensatus Rydb. КІ. Colo. 396. 1906, not Greene. S. mutabilis Nelson in Coulter & Nelson, Manual Cent. Rocky Mountains 583. 1909, mainly, not Greene. S. manitobensis Greenm. Ottawa Nat. 25:117. 1911, mainly. S. suavis Lunell, Am. Mid. Nat. 2:125. 1911. S. Metcalfei Greene, Contr. U. S. Nat. Herb. 16:193. 1913; Wooton & Standley, Contr. U. S. Nat. Herb. 19:748. 1915. S. remifolius Wooton & Standley, Contr. U. S. Nat. Herb. 16:194. 1913. An herbaeeous perennial, glabrous or somewhat white- floeceose-tomentulose and later glabrate except at the base and in the leaf-axils; stems one to several, frequently rather [Vor. 8 180 ANNALS OF THE MISSOURI BOTANICAL GARDEN densely tufted, 1 to 3 dm. high; lower leaves oblanceolate, entire, dentate towards the apex only or even pinnatisect, gradually narrowed at the base into a slender petiole, thick and firm in texture, including the petiole 2.5 to 10 em. long, .5 to 1 em. broad; upper stem-leaves becoming sessile and much reduced ; inflorescence a terminal few to several-headed corymbose eyme; heads 8 to 10 mm. high, radiate; involucre eampanulate, sparingly calyculate; bracts of Ше involucre 13 to 21, linear-lanceolate, 6 to 8 mm. long, glabrous; ray- flowers 10 to 12, rays yellow; disk-flowers numerous; achenes usually hirtellous on the ribs. Distribution: Manitoba to Texas, and west to Nebraska, Colorado, and New Mexico. Specimens examined: Manitoba: sand hills at Brandon and Old Wives Lakes, 22 June, 1887, Macoun 22 (Gray Herb.); on open prairie, south of Sewell, 12 June, 1876, Macoun (Geol. Surv. Canada Herb. 12232); gravelly or rocky places, Flat Creek, “N. W. T," 20 June, 1880, Macoun 103 (Geol. Surv. Canada Herb. 14796) ; Stewarts Lake Mountain, 21 June, 1875, Ma- coun 14777 in part (Geol. Surv. Canada Herb.) ; north of Car- berry, 14 June, 1906, Macoun £ Herriot (Geol. Surv. Canada Herb. and Field Mus. Herb.); without definite locality, coll. of 1898, E. S. Thompson (Mo. Bot. Gard. Herb.). North Dakota: on sand hills, MeHenry Co. 13 July, 1899, Lunell 24 (Gray Herb.). South Dakota: Hot Springs, Fall River Co., 6 June, 1893, Schenck (Mo. Bot. Gard. Herb.). Nebraska: Long Pine, 14 May and 4 June, 1893, Rutter (U. S. Nat. Herb.) ; sand hills on Middle Fork, Loup River, near Thedford, Thomas Co., 15 June, 1893, Rydberg 1311 (Gray Herb. and U. S. Nat. Herb.) ; in dry sandy soil, Hal- sey, 1 and 2 June, 1903, Мей Ф Knopf (Mo. Bot. Gard. Herb.); Hershey, 20 May, 1903, Mell 59 (U. S. Nat. Herb.) ; Franklin, coll. of 1893, Labourne (Mo. Bot. Gard. Herb.); Loup Fork, Hayden (Mo. Bot. Gard. Herb.). 1916] GREENMAN—MONOGRAPH OF SENECIO 181 Oklahoma: low places on prairie, near Camp, 12 May, 1913, Stevens 420 (Mo. Bot. Gard. Herb.) ; without definite locality, Stevens 489, 511 (G. W. Stevens Herb.). Texas: on prairies, Amarillo, 19 May, 1902, Reverchon 2523, 3330 (Mo. Bot. Gard. Herb.); prairies near Cañon City, June, 1901, Eggert (Mo. Bot. Gard. Herb.); Davis Mountains, 29 April, 1902, Tracy & Earle 336 (Gray Herb., U. S. Nat. Herb., Greene Herb., and Mo. Bot. Gard. Herb.) ; “mountains beyond Ше Limpia," Expedition from western Texas to El Paso, Мау-Осіоһет, 1849, Wright 403 (Gray Herb.), TYPE. Wyoming: open woods, Laramie Hills, Albany Co., 25 June, 1903, 4. Nelson 8962 (Mo. Bot. Gard. Herb.). Colorado: near Greeley, June, 1908, E. L. Johnston 461, 463, 466 (Mo. Bot. Gard. Herb.); dry plains, near Evans, eolls. of 1907—1909, E. L. Johnston 423, 423a, 424, 442, 443, 537, 538, 551, 552 (Mo. Bot. Gard. Herb.); Evans, 8 June, 1912, E. L. Johnston 804 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); in river bottoms, Fort Lupton, 19 May, 1913, E. L. Johnston 882, 883, 875 (U. S. Nat. Herb.) ; Clear Creek, Hay- den's U. S. Geol. Survey, 20 May, 1873, Coulter (Phil. Acad. Nat. Sei. Herb.), іп part; Lat. 39-419, coll. of 1864, Parry (U. S. Nat. Herb. 349250) ; Lat. 39-41°, coll. of 1862, Hall & Harbour 333 in part (Gray Herb. and Field Mus. Herb.); Denver, May, 1881, B. H. Smith (Phil Acad. Nat. Sci. Herb.); Denver, May, 1889, Eastwood (U. 5. Nat. Herb.) ; Denver, 8 June, 1891, E. C. Smith (Mo. Bot. Gard. Herb.) ; Denver, Williamson (C. S. Williamson Herb.); dry ground, Middle Creek, July, 1892, Beardslee 60 (U. S. Nat. Herb.) ; near Breckenridge, Summit Co., alt. 2950 m., Mackenzie 112 (Mo. Bot. Gard. Herb.); pasture land, Como, alt. 3050 m., 1 Aug. 1895, Crandall & Cowen 280 (0. S. Nat. Herb.) ; Leadville, H. A. Keller (Phil. Acad. Nat. Sei. Herb.) ; Lead- ville, 8 July, 1886, Trelease (Mo. Bot. Gard. Herb.) ; wet sandy places, along Cottonwood Creek, Buena Vista, Chaf- fee Co., alt. 2425-2440 m., 4 July, 1892, Sheldon 161, 479 (U. S. Nat. Herb.); open gravel bank, near railway station, [Vor. 3 182 ANNALS OF THE MISSOURI BOTANICAL GARDEN Limon, Lincoln Co., 13 June, 1912, Churchill (J. R. Churchill Herb.) ; Colorado Springs, May, 1879, M. E. Jones (U. 8. Nat. Herb. 223024 in part); Colorado Springs, alt. 1830 m., 8 Мау, 1897, А. А. 4 Е. G. Heller 3508 (Gray Herb., U. 8. Nat. Herb., and Mo. Bot. Gard. Herb.); Colorado Springs, May, 1892, Mulford (Mo. Bot. Gard. Herb.); Colorado Springs, 2 Aug., 1891, collector not indicated (Mo. Bot. Gard. Herb.); Pike's Peak, 10 July, 1901, Williamson (C. S. Wil- liamson Herb.); plains near Westcliffe, Custer Co., 25 July, 1887, Demetrio (Gray Herb.); Colorado Springs, 10 May, 1882, Allen & Brewster (U. 5. Nat. Herb. and Gray Herb.) ; saline soil on the plains, 12 Мау, 1870, Greene (Gray Herb.) ; Veta Pass, Sangre de Cristo Range, alt. 2740-3350 m., 9-16 June, 1890, Mr. Mrs G. H. Hicks 9 (Gray Herb.) ; Veta Pass, 15 July, 1896, Sheldon 3622 (U. S. Nat. Herb.) ; Pagosa Springs, 3 June, 1883, B. H. Smith (Phil. Acad. Nat. за. Herb.); southwestern Colorado, Hayden's U. 5. Geol. Survey, 1875, Brandegee (Mo. Bot. Gard. Herb.) ; wet moun- tain valley, July, 1885, Kettler (Gray Herb.) ; in dry fields, Manoos, alt. 2135 m., 8 July, 1898, Baker, Earle € Tracy 446 (Mo. Bot. Gard. Herb.); Rocky Mountains, coll. of 1888, Tracy (U. S. Nat. Herb. 48706). Nevada: eastern Nevada, coll. of 1883, Meehan (Phil. Acad. Nat. Sel. Herb.). New Mexico: dry hills, vicinity of Raton, Colfax Co., alt. 2100-2380 m., 21-22 June, 1911, Standley 6278 (U. S. Nat. Herb.); upland slopes, Catskill, June-July, 1895, Mrs. О. St. John 139 (Gray Herb.); plains on and near the Sierra Grande, Union Co., alt. 2100-2935 m., 19 June, 1911, Standley 6123 (U. S. Nat. Herb.); along the river and in damp meadow, vicinity of Chama, Rio Arriba Co., alt. 2380-2850 m., 8 and 9 July, 1911, Standley 6532, 6734 (U. S. Nat. Herb.); pass southeast of Tierra Amarilla, Rio Arriba Co., alt. 2300 m., 18 April-25 May, 1911, Eggleston 6531, 6595 (U. S. Nat. Herb.) ; Hillsboro Peak, Grant Co., alt. 3100 m., 27 May, 1904, Metcalfe 938 (U. S. Nat. Herb., Gray Herb., and Mo. Bot. Gard. Herb.), түре of S. Metcalfei; Willow 1916] GREENMAN—MONOGRAPH OF SENECIO 183 Creek, 8 Aug. 1908, Wooton (U. S. Nat. Herb.), түре of S. remifolius. 78. S. Wardii Greene, Pittonia 4:116. 1900; Heller, Cat. N. Am. Р]. 231. 1900. РІ. 3, fig. 3. A low herbaceous perennial, glabrous or slightly tomentu- lose in the leaf-axils; stems tufted, erect, less than 1 dm. high; lower leaves oblong-oblanceolate including the petiole l5 to 4 em. long, 3 to 10 mm. broad, entire or dentate towards the obtuse or rounded apex; stem-leaves few, more or less braeteiform; inflorescence a terminal round-topped rather dense есуше; heads 6 to 8 mm. high, radiate; involucre campanulate, sparingly calyculate, glabrous or nearly so; bracts of the involucre about 13, linear-lanceolate, 4 to 5 mm. high; ray-flowers 8 to 10, rays yellow; disk-flowers numerous; achenes glabrous. Distribution: high mountains of Utah. Specimens examined: Utah: Fish Lake Mountain, and Thousand Lake Mountain, alt. about 3500 m., U. S. Geol. and Geog. Survey of the Ter- ritories, 8 July, 1875, Ward 332 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.), түре. This species may be looked for in herbaria under S. aureus var. alpinus un- der whieh name it was distributed. 79. S. anacletus Greene, Pittonia 4:307. 1901; Rydb. FI. Colo. 395. 1906; Coulter & Nelson, Manual Cent. Rocky Moun- tains 579. 1909. Pl. 4. S. microdontus (Gray) Heller, Bull Torr. Bot. Club 24:479. 1897, not Baker; Cat. N. Am. Pl. 146. 1898, and ed. 2, 230. 1900; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902; Wooton & Standley, Contr. U. S. Nat. Herb. 19:746. 1915. S. Toluccanus var. microdontus Gray, Syn. Fl. N. Am. 1? :388. 1884, and ed. 2, 1886. S. Wootonii Greene, Bull. Torr. Bot. Club 25 :122, pl. 331, figs. 1, 2. 1898. An herbaceous perennial, glabrous throughout and usually glaucous; stems one to several from an ascending fibrous- [Vor. 8 184 ANNALS OF THE MISSOURI BOTANICAL GARDEN rooted rather stout rootstoek, simple or branched, ereet, 2 to 5 dm. high; lower leaves obovate to oblong-oblanceolate, including the petiole D to 2.5 dm. long, .5 to 4.5 em. broad, thick and firm in texture, rounded to submucronate-acute at the apex, entire to sinuate-denticulate, gradually narrowed at the base into a slightly winged petiole; stem-leaves few, oblanceolate to lance-attenuate, sessile and entire; inflo- rescence terminating the stem and branches in a simple or compound согутђоѕе cyme; heads 10 to 12 mm. high, radiate; involucre campanulate, calyculate; bracts of the involucre about 13, linear-lanceolate, 7 to 9 mm. long, acute, glabrous except at the brownish penicillate tips; ray-flowers 8 to 10, rays yellow; disk-flowers numerous; achenes striate, glabrous. Distribution: Colorado to northern Mexico. Specimens examined: Colorado: pine woods below Berthoud’s Pass, alt. 3200 m., 10 Aug., 1874, С. Engelmann (Мо. Bot. Gard. Herb.) ; sum- mit of steep mountains, Estes Park, alt. 3050 m., 7 July, 1912, Churchill (J. R. Churchill Herb.) ; Tolland, alt. 2740 m., 23 July, 1913, Overholts (Mo. Bot. Gard. Herb.) ; damp places in the valley near Empire, alt. about 3050 m., 21 July, 1892, Patterson 199 (Gray Herb.); valley near Empire, alt. 2590 m., July, 1892, Patterson 202 (Mo. Bot. Gard. Herb.) ; Breckenridge, coll. of 1887, Bereman (Mo. Bot. Gard. Herb.); Redeliff, Eagle Co., 17 July, 1902, Osterhout 2704 (Phil. Acad. Nat. Sci. Herb.) ; Leadville, 6 July, 1886, Tre- lease (Mo. Bot. Gard. Herb.) ; ‘‘Sawatch’’ Range, alt. 3350 m., Aug., 1880, Brandegee (Gray Herb. and Mo. Bot. Gard. Herb.); head waters of Clear Creek, ete., coll. of 1861-62, Parry 21 (Gray Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.); Lat. 39-419, Hall & Harbour 326 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.); Taylor River, alt. 2895 m., Hayden's U. S. Geol. Survey, 3 Aug., 1873, Porter (U. S. Nat. Herb.) ; meadows, South Cottonwood Gulch, Chaffee Co., alt. 3200 m., 9 July, 1892, Sheldon 169, 488 (U. S. Nat. Herb.) ; Silverton, 1916] GREENMAN—MONOGRAPH OF SENECIO 185 July, 1889, Eastwood (U. S. Nat. Herb.); Silverton, alt. about 2895 m., 3 and 16 July, 1898, ex Herb. Colorado State Agr. Coll. 3097, 3117 (U. S. Nat. Herb.); Pagosa Springs, 4, June, 1894, B. H. Smith (Phil. Acad. Nat. Sei. Herb.) ; Little Kate Mine, La Plata Mountains, alt. 3352 m., 16 July, 1898, Baker, Earle £ Tracy 552 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; La Plata River, 16 July, 1898, Baker, Earle & Tracy 994 (Mo. Bot. Gard. Herb.) ; Rocky Mountains, coll. of 1868, Vasey (U. S. Nat. Herb.). New Mexico: pass south of Tierra Amarilla, Rio Arriba Co., alt. 2320 m., 18 April-25 May, 1911, Eggleston 6530, 6617 (U. S. Nat. Herb.); pine woods, vicinity of Brazos Cañon, Rio Arriba Co., 21 Aug., 1914, Standley € Bollman 10676 (U. S. Nat. Herb.); Rio de la Casa, above Mora, 30 May-1 June, 1902, Sturgis (Gray Herb.) ; valley of Santa Fé Creek, at the foot of mountains, ten miles above Santa Fé, coll. of 1847, Fendler 477 (437) (Gray Herb. U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb, and Mo. Bot. Gard. Herb.); Sante Fé Cafion, nine miles east of Santa Fé, alt. 2440 m., 2 June, 1897, 4. A. Ф E. G. Heller 3648 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.) ; Hermit's Peak, San Miguel Co., Aug., 1884, F. H. Snow (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Rio Pecos, three miles above Winsor's, alt. about 2650 m., 30 June, 1908, Standley 4083 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.) ; White Moun- tains, Lincoln Co., alt. 2130 m., 15 Aug., 1897, Wooton (U. S. Nat. Herb., Greene Herb., fragment in Mo. Bot. Gard. Herb.), TYPE of S. Wootonii; White Mountains, Lincoln Co., alt. 2255 m., 25 Aug., 1907, Wooton & Standley 3510 (U. S. Nat. Herb.) ; Cloudcroft, Otero Co., alt. 2590 m., 29 May-5 June, 1902, Viereck (Phil. Acad. Nat. Sci. Herb.) ; James Cañon, Sacra- mento Mountains, Otero Co., 26 June, 1899, Wooton (U. S. Nat. Herb.); Pinos Altos Mountains, 21 May, 1880, E. L. Greene (Gray Herb.); Pinos Altos, 15 Aug., 1895, Mulford 881 (Mo. Bot. Gard. Herb.). Arizona: San Francisco Mountains, coll. of 1884, Lemmon 3264 (Gray Herb.); San Francisco Mountains, alt. 2135- [Уо 3 186 ANNALS OF THE MISSOURI BOTANICAL GARDEN 2440 m., 22 June, 1891, McDougal 253 (U. S. Nat. Herb.) ; Kendrick Peak, near Flagstaff, May—Oct., 1900, Purpus 8001 (Mo. Bot. Gard. Herb.); near Flagstaff, alt. 1850 m., 16 Aug., 1901, Leiberg 5859 (U. S. Nat. Herb.) ; Apache-Verde, east of Baker Butte, Black Mesa Forest Reserve, 1 June, 1900, Coville 1040 (U. S. Nat. Herb.) ; Santa Catalina Moun- tains, alt. 2740 m., April, 1881, Lemmon 190 (Gray Herb.) ; Rincon Mountains, alt. 2285 m., coll. of 1891, Neally 221 (U. S. Nat. Herb.); Santa Rita Mountains, alt. 2135-2440 m., 3 May, 1881, Pringle (Gray Herb., U. S. Nat. Herb., and Phil. Acad. Nat. Sci. Herb.); in pine forests, Santa Rita Mountains, alt. 2285 m., 8 June, 1884, Pringle (Phil. Acad. Nat. Sei. Herb.); Tanner's Cañon, near Fort Huachuca, coll. of 1882, Lemmon 2786 (Gray Herb.); Willow Spring, 10-20 June, 1890, Dr. E. Palmer 479 (Gray Herb. and U. S. Nat. Herb.) ; Ramsey Canon, Huachuca Mountains, 10 April, 1915, Blumer 5929 (Mo. Bot. Gard. Herb.); rolling andesitie pine land, Barfoot Park, Chiricahua Mountains, alt. 2440-2500 m., 17 Sept., 1906, Blumer 151, 1388 (U. 8. Nat. Herb. and Mo. Bot. Gard. Herb.). Chihuahua: Colonia Gareia in the Sierra Madres, alt. 2285 m., 6 June, 1899, Townsend Ф Barber 14 (Gray Herb. and Mo. Bot. Gard. Herb.). 80. S. toluccanus DC. Prodr. 6:428. 1837; Schz. Bip. in Seemann, Bot. Voy. Herald 311. 1852-57; Hemsl. Biol. Cent.- Am. Bot. 2:248. 1881; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. РІ. 5. An herbaceous perennial; stems 2 to 10 dm. high, erect or ascending from a thick stout rootstock; lower leaves ovate to oblong-oblanceolate, including the petiole 5 to 30 cm. long, 1 to 10 em. broad, acute or obtuse, slightly cre- nate to conspicuously dentate with spreading subear- tilaginous teeth, narrowed at the base into a winged petiole, thiek and firm in texture, glabrous on both surfaces or slightly tomentulose in the early stages and soon glabrate; stem-leaves few, sessile and semi-amplexieaul, lanceolate, dentate to entire; inflorescence terminating the stem in a 1916] GREENMAN—MONOGRAPH OF SENECIO 187 few to many-headed glabrous or somewhat pubescent corym- bose eyme; heads 10 to 15 mm. high, radiate; involucre cam- panulate, calyculate; bracts of the involucre 13 to 21, linear- lanceolate, 7 to 9 mm. long, glabrous or nearly so; ray- flowers 10 to 12, rays yellow; disk-flowers numerous; achenes striate, glabrous. Distribution: Mexico. Specimens examined: Coahuila: south of Saltillo, Feb.-Oct., 1880, Dr. E. Palmer (Gray Herb.). San Luis Potosi: Valley of San Luis Potosi, Aug., 1876, Schaffner 277 (Gray Herb. and U. S. Nat. Herb.) ; Valley of San Luis Potosi, alt. 1830-2440 m., Parry & Palmer 537 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.). Jalisco: Nevada de Colima, alt. 3050-3650 m., 16 May, 1893, Pringle 4374 (Gray Herb., U. S. Nat. Herb., and Mo. Bot. Gard. Herb.). Michoacan: north slope of Mt. Patambau, alt. 2895-3350 m., 1-4 Feb., 1903, E. W. Nelson 6598 (U. S. Nat. Herb.). Mexico: Sierra de las Cruces, 11 Sept. 1892, Pringle 5261 (Gray Herb.); Nevada de Toluca, 15 Oct., 1903, Rose Ф Painter 7901 (Gray Herb.). Federal District: Serrania de Ajusco, alt. 3050 m., 16 April, 1898, Pringle 6812 (Gray Herb., U. S. Nat. Herb., Phil. Acad. Nat. Sci. Herb., and Mo. Bot. Gard. Herb.). Southern Mexico: “cuesta de las papao pr. Angangueo”’, Feb., 1830, Schiede (Berlin Herb., Gray Herb., and U. S. Nat. Herb.); wet places on Mt. Ixtaecihuatl, alt. 2740-3050 m., Nov., 1905, Purpus 1509 (U. S. Nat. Herb. and Mo. Bot. Gard. Herb.); Mt. Orizaba, 21 April, 1893, E. W. Nelson 5 (U. S. Nat. Herb.) ; Mt. Orizaba, alt. 4025-4265 m., 18 March, 1894, Е. W. Nelson 281 (U. S. Nat. Herb.) ; wet grassy soil near timberline, Mt. Orizaba, March, 1908, Purpus 2976 (U. S. Nat. Herb.); ‘‘Vera Cruz,’’ Galeotti 2183 (Kew Herb.) ; at the base of Jacal (Joerl), June, 1839, Ehrenberg 1294 (Berlin Herb. and Gray Herb.) ; in woods near Guapi- [Vor. 3 188 ANNALS OF THE MISSOURI BOTANICAL GARDEN malpam, coll. of 1854, Schaffner (Gray Herb.) ; without defi- nite locality, coll. of 1848-1849, Gregg 690 in part (Mo. Bot. Gard. Herb.). Var. modestus Schz. Bip. in Seemann, Bot. Voy. Herald 311. 1852-57; Hemsl. Biol. Cent.-Am. Bot. 2:248. 1881; Greenm. Monogr. Senecio, I. Teil, 24. 1901, and in Engl. Bot. Jahrb. 32:20. 1902. Stems slender, 3 to 4 dm. high, nearly naked above; lower leaves oblanceolate, 3 to 14 em. long, .5 to 1.5 em. broad. Distribution: northwestern Mexico. Specimens examined: Northwest Mexico: Sierra Madre, Seemann (Kew Herb. and Gray Herb.). Chihuahua: Mound Valley, south of Pacheco, 12 June, 1891, Hartman 690 (Gray Herb. and Mo. Bot. Gard. Herb.). (To be continued.) [VoL. 3, 19161 190 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 3 Fig. 1. Senecio Rosei Greenm. exico From the type D Rose No. 2157, in the Gray Herbarium of Harvard University. Fig. 2. Senecio Wardii Greene h From a co-type MN Ward No. 332, in the Gray Herbarium of Harvard University. Fig. 3. Senecio hesperius Greene Oregon From Howell's No. 160 in the Gray Herbarium of Harvard Univer- sity. SE S ^ ANN. Мо. Вот. GARD., VOL. 8, 1916 PLATE 3 GREENMAN—MONOGRAPH OF SENECIO COCKAYNE, BOSTON. [Vor. 3, 1916] 192 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 4 Senecio anacletus Greene United States From Heller’s No. 3648 in the Herbarium of the Missouri Botanical Garden. ANN. Mo. BOT, GARD., VOL, 3, 1916 PLATE 4 GREENMAN—MONOGRAPH OF SENECIO COCKAYNE, BOSTON, 194 [Vor. 3, 1916] ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 5 Senecio toluccanus DC. Mexico From Pringle’s No. 6812 in the Gray Herbarium of Harvard Uni- versity. e E ж." 7 СКЕЕММАН--МОМОСКАРН OF SENECIO COCKAYNE, BOSTON. NEW OR INTERESTING SPECIES OF GILL FUNGI FROM MISSOURI L. 0. OVERHOLTS Formerly Rufus J. Lackland Fellow in the SE Shaw School of Botany of Washington Universit The following collections of gill ne made by the writer in the vicinity of St. Louis, Missouri, within the past two years are believed to be undescribed. Claudopus subnidulans Overholts, n. sp. Pileus sessile, reniform or dimidiate in outline, convex, 0.5-2 em. broad, bright tawny orange, fibrillose-tomentose, dry; margin inrolled, even or very slightly striate; flesh thin, white; odor and taste none; gills radiating outward from the point of attachment of the pileus, medium distant, rather broad (3-5 mm.), salmon-colored or dull orange; stem none, the pileus attached by a white tomentose base; spores salmon- colored, globose, smooth, 5-7» broad; cystidia none. On rotten logs in damp woods. Jefferson Barracks, near St. Louis, Missouri, October 25, 1913. Type collection in Herb. Overholts No. 1460, and врес!- mens from this collection are deposited in the herbarium of the Missouri Botanical Garden. The species resembles C. nidulans, but differs in the con- stantly smaller size and the exactly globose spores. Panaeolus reticulatus Overholts, n. sp. Plate 6, fig. 1. Pileus hemispheric then expanded, sometimes somewhat umbonate, 3.5-6 em. broad, between drab and light brownish umber when young, usually smoky brown or blackish with age, dry, everywhere pitted or when older appearing fibrous- recticulate; the margin even, at first incurved then wavy; flesh thin, watery, pallid; taste and odor not characteristic; gills broadly attached but sometimes sinuate, often separat- ing with age, spotted, or in age uniformly black, rather close, ANN. Mo. Вот. GARD., Vor. 3, 1916 (195 [Vor. 3 196 ANNALS OF THE MISSOURI BOTANICAL GARDEN unequal, 4-7 mm. broad, the edge whitish; stem central, terete, equal or slightly tapering downward, hollow, twisted, pruin- ose-scabrous at the apex, somewhat shining, pale brown, car- tilaginous and brittle, 4-8 em. long, 4-8 mm. thick; veil not apparent; spores elliptic or broadly elliptic, nearly black, 8-10Х5.5-6.5и; cystidia none. Cespitose or gregarious on earth in flower beds in the Mis- souri Botanical Garden, May 31, 1915; also at the same place, June 17, 1915. Type collection in Herb. Overholts No. 2795, and specimens from this collection are deposited in the herbaria of the New York Botanical Garden and the Missouri Botanical Gar- en. The species is in every way distinct from P. retirugis Fries and P. alveolatus Peck. The pileus when very young is pitted only in the center, but mature plants are pitted all over and at times appear reticulate. When dried the pileus is smooth or nearly so. Panaeolus rufus Overholts, n. sp. Plate 6, fig. 2. Pileus convex to nearly plane, sometimes broadly umbonate, 2-5 em. broad, varying in color from tan to light brown or chestnut, darkest at the center, dry, glabrous, often becoming cracked and areolate except at the center; margin even, ex- tending slightly beyond the lamellae; flesh thin, white; odor none; taste farinaceous; gills adnate or adnexed, medium close or slightly distant, spotted, becoming blackish brown, whitish on the edge; stem central, terete, pruinose, striate, light-colored above, becoming dark reddish brown below, white tomentose at the base, firm, cartilaginous, hollow, 5-10 em. long, 2.5-6 mm. thick; veil not apparent; spores elliptic or broadly elliptic, almost black in mass, 12-13,5Х7.5-9и; cystidia none. Gregarious on a compost heap in the Missouri Botanical Garden, June 1, 1915. Type collection in Herb. Overholts No. 2796, and вресі- mens from this collection are deposited in the herbaria of the Missouri Botanieal Garden and the New York Botanical Garden. 1916] OVERHOLTS—GILL FUNGI 197 The plants are more highly colored than in any described species of Panaeolus. Panaeolus variabilis Overholts, n. sp. Plate 6, figs. 3, 4. Pileus slightly campanulate to convex or plane, young speci- mens indistinctly umbonate, 2-6 cm. broad, very variable, when young hygrophanous, fleshy brown mingled with gray, somewhat rugose, when mature dry and lighter or creamy white, glabrous; margin even; flesh thin, concolorous; odor none; taste slightly farinaceous; gills adnate to adnexed, at first light brown, then spotted, finally black, rather close, 3-6 mm. broad, whitish on the edge; veil none; stem central, terete, equal or nearly so, pallid to slightly flesh-color or dark brown, floccose-pruinose when young, usually striate at the apex, hollow from the first, 4-9 em. long, 2-5 mm. thick; spores broadly elliptic or ellipsoid, black, 12-13X 7-95; cystidia none. Gregarious or subcespitose on earth in flower beds and among shrubbery in the Missouri Botanical Garden, May 31, 1915; also from the same place, June 17, 1915. Type collection in Herb. Overholts No. 2794, and specimens from this collection are deposited in the herbaria of the Missouri Botanical Garden and the New York Botanical Gar- den. The species is a very variable one. AN INTERESTING VARIETY OF PLUTEUS CERVINUS In November, 1914, while collecting in the region of Pa- cific, Missouri, the writer found a large cluster of a species of Pluteus growing in the sawdust on an old sawmill site. There were about thirty individuals in the cluster, and they ranged from 11 to 16 em. broad. The specimens were much larger than is usual in Pluteus cervinus, and the fibrils on the pileus were much more conspicuous than in that species. These facts, together with the cespitose habit and another character mentioned below, seemed to justify the separation of these specimens into a new species. Further study has led the writer to modify this first conclusion, and the plants are now referred to P. cervinus. The variations are so marked, however, that they deserve notice, the accompanying [Vor. 3 198 ANNALS OF THE MISSOURI BOTANICAL GARDEN figures showing these points. For convenience this collection will be referred to below under my herbarium number, 2316. A microscopic examination of the hymenium of this and other collections of P. cervinus reveals some interesting va- riations in the form of the cystidia. These are more or less flask-shaped structures and hyaline. The accompanying fig- ures show the variations. In all collections examined (ex- cept the one referred to above) some of the eystidia have peeuliar thorn-like projections more or less abundant. Figure A is from my herbarium, No. 2809, and in most eollections it is probably the most typical form present. Figures В, С, and D are from my herbarium, No. 1624, and Ше thorny type ts A в c Fig. 1. Various types of eystidia found in hymenium of Pluteus cervinus: A, from herbarium No. 2809; B, C, and D, from herbarium No. 1624, the thorny type D being most abundant; E, eystidium of P. cervinus var. caespitosus. (This type is also present in other col- leetions.) F, spores of P. cervinus; G, spores of P. cervinus var caespitosus. of eystidium was better developed in that collection than in any other one examined. These sharp projections were most often not present on the apex of the cystidium, but were scat- tered along the sides as thorns on a stem. In both eollections eited and in all others examined there were present also a large number of entirely smooth, sharp-pointed eystidia. When collection No. 2316 was examined no cystidia with thorny projections were seen. It 15 very doubtful whether this is a constant character on which, together with other before- mentioned facts, a new species might be segregated. How- ever, there are certainly no such eystidia present as in the other collections; but the presence in other collections of en- 1916] OVERHOLTS—GILL FUNGI 199 tirely smooth cystidia seems to bridge over the gap in this respect, and it seems best at present to regard this latest collection as only a variety of Pluteus cervinus. P. petasatus Fries, sometimes regarded as a synonym of P. cervinus, approaches this collection in size and is reported as growing on sawdust, but it is described as glabrous and commonly umbonate—characters that do not apply to my plants. For convenience the present collection may be desig- nated as follows: Pluteus cervinus var. caespitosus Overholts, n. var. Plate 6, figs. 5, 6. Pileus 11-16 em. broad, very slightly viscid, decorated with brownish fibrils or appressed, fibrillose scales that are more prominent in the center; gills 1-1.7 em. broad; stem 10-15 em. long, 1.3—2.5 em. thick; spores oblong-ellipsoid, smooth, hyaline under the microscope, salmon-colored in mass, 4-7 Х3- 4»; cystidia abundant, fusiform, sharp-pointed, smooth, 40— 75 X 10-12, | On heap of sawdust. Densely cespitose in a cluster of about thirty plants. Pacific, Missouri, November 9, 1914. Type collection in Herb. Overholts No. 2316, and a single specimen from the collection has been deposited in the her- barium of the Missouri Botanical Garden. The variety is edi- ble and quite delicious. [ Vor. 3, 1916 200 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 6 Fig. Panaeolus reticulatus. From photograph of type specimen in Her- Ее Overholts No. 2795. Panaeolus rufus. From photograph of type specimen in Herbarium пе. 2 No. 2796. Figs.3 and 4. Pamaeolus variabilis. From photographs of type specimen in қ җа бын Overholts No. 2794. Figs.5 and 6. Pluteus cervinus var. caespitosus. From photographs of type specimen in elei Overholts No. 2316. ANN. Мо. Вот. GARD., Vor. 3, 1916 PLATE 6 e — OVERHOLTS— GILL FUNGI Annals of the Missouri Botanical Garden Vor. 3. APRIL, 1916 No. 2 A NEW SENECIO FROM JAMAICA? J. M. GREENMAN Curator of the Herbarium of the Missouri Botanical Garden Associate Professor in the Henry Shaw School of Botany of Washington University In the spring of 1908 Dr. N. L. Britton and Dr. Arthur Hollick discovered in the Parish of St. Ann’s, Jamaica, a vine-like Senecio which appeared to them to be different from any species recorded in this genus. The plant was des- ignated temporarily by Dr. Britton as ‘5. НоШски, n. вр.” and generously submitted to the writer for examination. From the limited material available at the time the author was unable to find sufficient characters to separate it satis- faetorily from S. Swartz DC. Additional specimens have been secured since, and further information acquired as to the habit of the two plants concerned. The writer takes pleasure in confirming Dr. Britton’s view and in placing on record the following description: Senecio Hollickii Britton, sp. nov. Caulis lignescens scandens usque ad 6.5 m. longus cortice griseo tectis; ramis floriferis teretibus striatis brunneis gla- bris vel paree pubescentibus; foliis alternis petiolatis ovatis vel laneeolatis 2.5-12 em. longis 1.5—4 em. latis superne sen- sim angustatis acutis integris vel remote subdenticulatis utrinque glabris subtus pallidioribus basi acutis vel subeor- datis, margine plus minusve revolutis; petiolis ] em. vel minus longis glabris; inflorescentiis terminalibus cymosis 1Issued September 30, 1916. ANN. Mo, Bor. GARD., Vor. 3, 1916. (201) [VoL. 3 202 ANNALS OF THE MISSOURI BOTANICAL GARDEN parce pilosulis multicapitatis; capitulis circiter 1 em. altis heterogamis; involucris subcylindratis ealyeulatis; involucri squamis plerumque 8 lineari-lanceolatis 7-8.5 mm. longis acutis glabris penicillatis; floribus femineis 4 ligulatis, ligulis oblongo-elliptieis 4-5 mm. longis 2.5 mm. latis, aurantiabus; floribus disei plerumque 7; pappi setis albis; achaeniis stri- atis superne pilosis. Specimens examined: Jamaica: rocky hillside, Union Hill, near Moneague, Par- ish of St. Ann's, alt. 450 m., 6-7 April, 1908, Britton Ф Hol- lick 2729 (N. Y. Bot. Gard. Herb., photograph and fragment in Mo. Bot. Gard. Herb.), түре; Pramble, near Claremont, alt. 520 m., 22 Jan., 1898, Fawcett € Harris 7032 (N. Y. Bot. Gard. Herb.); Soho, St. Ann, alt. 425 m., 11 May, 1915, Har- ris 11983 (N. Y. Bot. Gard. Herb.). The species here described, namely S. Hollicku, differs from S. Swartzii to which it is perhaps most closely related in inflorescence, involucre and floral characters, in being a vine instead of a shrub or tree, in having ovate or lanceolate instead of oblong or obovate leaves, in having a more con- spicuous venation with the lateral nerves less divaricately spreading and thus forming a more acute angle with the midrib, in having shorter petioles, and finally in having pubescent instead of glabrous achenes. Mr. William Harris states, with reference to his No. 11983 from Soho, St. Ann, that it is a plant ‘‘climbing over shrubs and trees to a height of twenty feet.” THE THELEPHORACEAE OF NORTH AMERICA УІ Hypocunvus EDWARD ANGUS BURT Mycologist and Librarian to the Missouri Botanical Garden Associate Professor in the Henry Shaw School of Botany of Washington University HYPOCHNUS Hypochnus Fries emend. Karsten, Rev. Мус. 3:23. 1881; Finska Vetenskaps-Soc. Bidrag Natur och Folk 37:162. 1882; Finl. Basidsv. 438. 1889; Fries, Obs. Mye. 2:278. 1818 and 1824, (in part); Syst. Мус. 3:289. 1829, (in part); Gen. Hym. 16. 1836, (in part); Еріст. 569. 1838, (in part); Басс. Syll. Fung. 6:653. 1888, (in part) ; R. Fries, R. Sci. Soc. Goth- oburgens Actis IV. 3:37. 1900. — Hypochnus as a subgenus of Corticium Fries, Hym. Eur. 659. 1874, (in part). — To- mentella Persoon ex Patouillard? Hym. Eur. 154. 1887; Schroeter, Krypt.-Fl. Sehlesien 3:419. 1888; Engl. & Prantl, Nat. Pflanzenfam. (I:1**) :117. 1898. — T'omentellina v. Höh- nel E Litschauer, K. Akad. Wiss. Wien Sitzungsber. 115: 1604. 1906. Fruetifieations resupinate, effused, dry, coriaceous, felt- like or hypochnoid, usually composed of loosely interwoven hyphae which bear basidia sometimes in scattered clusters but more usually in a compact hymenium; hymenium even or papillose; basidia simple, bearing two or more spores, rough-walled to echinulate, distinctly colored in most species, pale-colored in a few, and hyaline in one or possibly more species. 'Issued September 30, 1916. ?Patouillard and аве in the works cited above, — Tomen- tella to Гене. because ће used this word in pu а in the names of two рү in his pu ublished peres] Obs. Mye. 2:18 and 19, 1799, as Elton: *97. Corticium (Tomentella) f errugineum. **98. Corticium ( Tomentella) chalibeum.’ s not generic ғана of Tomentella. Why Persoon used the word is not evident; he A not adopt it as a genus in his following formal works: ` Syno s Fungorum’ published in 1801, and ‘Mycologia ' jn 1822. Erbe y publiestion of Tomentella was not made until 1887 by Patouillard six years after Karsten's emendation of Hypochnus; hence € a fo, Вот. GARD., Vor. 3, 1916 (203) [Vor. 3 204 ANNALS OF THE MISSOURI BOTANICAL GARDEN Hypochnus is separated from Thelephora, as I have limited the latter, by strictly resupinate habit; from Corticium and Pemophora by rough-walled to echinulate spores which are usually, but not always, distinetly colored; from Zygodesmus of the Hyphomycetes by true basidia which bear two or more spores; and from Grandinia and Odontia of the Hydnaceae by loosely interwoven, hypochnoid structure and more or less colored, rough-walled to echinulate spores. As here treated, the species of Hypochnus form a natural, compact group at the foot of Hymenomycetes, with simple basidia, and closely resembling Zygodesmus in general habit and also in form and color of spores. Hypochnus is so closely related to Thelephora and Grandinia that many of its species have been published in those genera, as will be seen by the synonymy of species, or occur in those genera under manuseript names in the large herbaria. The species of Hypochnus are apparently humus formers, for the fructifications are found under very rotten wood and other organie matter rather than on nearly sound wood. Hence they probably follow other fungi in wood destruc- tion. This is the first presentation of the North American species of Hypochnus. It shows the geographical distribution of the genus localized in the northeastern United States and along our Atlantie coast and ranging westward aeross the northern United States. Not an Hypochnus has been found in a series of 175 numbers of Thelephoraceae, mostly re- supinate, collected by Dr. and Mrs. Murrill in Mexico. The sketches of microscopic details of the species in this part were made by the aid of a camera lucida from prepara- tions of such type or authentic specimens as are referred to in the accompanying text. The development of the present conception of Hypochnus is of historical interest. When first published, Hypochnus comprised species which I refer to Hypochnus and Corticium; then tropical lichens predominated; in his last work Fries excluded the lichens, recognized the close relationship to Corticium and placed both Coniophora and Hypochnus as 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 205 subgenera of Corticium. As several species of Corticium were still included in Hypochnus, Fries had good reason for regarding Hypochnus in his sense as closely related to Cor- исшт. Karsten’s emendation of Hypochnus a few years later was logical, and in sympathy with the work of Fries, for it retained this name for the greatest number of co- generic species both originally published in the genus and retained in the final work of Fries. These species are fur- thermore the only species for which the generic name Hy- pochnus can be retained, for the other species of the sub- genus in Fries’ ‘Hymenomycetes Europeae’ revert to Cor- ticium under modern study. Hypochnus, as presented in Saccardo’s ‘Sylloge Fungo- rum,’ is an aggregation of species of several genera and includes also the tropical lichens which Fries excluded from the genus in 1874. Hypochnus as given in Engler & Prantl’s ‘Die Natiirlichen Pflanzenfamilien,’ is the presentation of a purely academic scheme of Schroeter’s as to how the lower Hymenomycetes ought to be classified to have a family Hypochnacei, but the fungi do not fall in with the scheme. They cannot be separated from Corticium and Peniophora. Von Hohnel and P. Sydow have pointed out! that Hypochnus in the sense of Schroeter must be abandoned as a genus and its species take their proper places in other genera. It is to be regretted that Saccardo’s ‘Sylloge Fungorum’ and Engler & Prantl’s ‘Die Natürlichen Pflanzenfamilien’ give a false lead with regard to Hypochnus, for these works are the main reliance of plant pathologists in the matter of genera. Ккү то THE SPECIES Spores distinctly colored as seen with the microscope ................. Spores e? ES yellowish or hyaline as to appear hyaline or nearly inder the microse соре......................аЖ ee 1 Fruetification {етти inous,’’ i. e, Sudan-brown,* Brussels-brown, = hazel of Ridgway spores concolorous with the fructification, but w yellow under E mieros eope er HESS cR Lo 1Ann. Myc. 4:551. 1906. See also von Hóhnel & Litschauer, Ann. Myc. 4:288. 1906. e technical color terms used in this work are those of Ridgway, Color Standards and Nomenclature. Washington, D. C., 1912. [Vor. 3 206 ANNALS OF THE MISSOURI BOTANICAL GARDEN 1. ее not ‘‘ferruginous’’; spores not wax-yellow under the КЕ. as pus reer re rrr Creer rrr ee REC HERE EE 2. "Without СТЕНО олоон ета еен зела еа КУЕ 3 2. n th deg? — of non-inerusted, cylindric organs protrud- ing from the Һутевйһиі......-.-....»»-..:....--.-. 4. H. canadensis 3. oe adnate, all Ge? E colored like the spores; RE echin- OC d EE H. ferrugineus all hyphae colored like the EEN 2. H. пречи Ga “Ж ні е Q 3 5 =. ET о EI et = o B т © SE E - = Dä С: — © + H о м 5 un с = un e = > Gi е Е өжет ; ubhy menial hyphae colored like the spores; spor s echin late 8. H subferruginens . nis ae not 2” 1. e, not having clamp connectio Hyphae nodose-septate, i. e., with clamp connections ............. ç Praci atin ranging a? drab to fuseous and Chastusa- drab, sepa- rable; spores and hyphae coneolorous, dark olive- buff to buffy brown under the microscope; hyphae 4-5и in diameter; ism ec Be ТОША: Lee ege ee RE H. wmbrinus e EE -brown beeoming Rood's brown, adnate; hyphae colored next to substratum, hyaline in subhymenium, 4—5y in diameter; spores Se phis the body 5-64 in diameter or 5- 645 e —————— Án age OOS жақа жасан 21. H. subvinosus я Wee dc _ деер olive-buff to dark olive-buff, adnate; spores and АМ зони color hyphae near the substratum 8- -10д, or more, іп diam géie echinulate the body 7-9а in diameter.............. 12. H. ppi ene 6. Without cystidia 7 6. With eystidi “madera A of non-inerusted cylindric жұн Pe ing B RO: Ze vs. ns e Eat v XC CC ER Ж, ие Margin of the e sime yel as p hymenial surfaee . Margin of differ = from the hymenial ѕитЁасе.................. 8. Tructifieation pou ored — einnamon-drab, DENM sepia, fuscous — and the hyphae са, PTE E E EE 8. Pruetiieation sepia or ising, and the hyphae 2. or hyaline nder the microscope after treatment with KHO solutio 8. Fructiieation rahi ng in brown from Saccardo’s i rig and snuff- bro amon-brown; hyphae and spores “x ig with the po Por s spores echinulate, the body 6-8 х5-7ш.......... BEE "18. ре: pannosus 8. pon between cartridge-buff and olive-buff; hypha and spores snuff-brown under the mieroseope; known from Washi ing- 0 gie Tr ХУ 14. H. avellaneus 8. pam drab or gray, and the hyphae hyaline under the mi- ; Fruetifieation with distinct vinaceous tinge, 250-3504 thick; hyphae suberect, ech? rou beet ее Sr pat ша I than the spores under the spores aeulea r echinulate.......... H. fuscus 7 Froctification Pepe? es Saccardo’ 8 m ш. NEE rarely бота 200-1200u thick; hyphae thick-walled, not rough- walled, extending in sca ioca in the subieulum и loosely interwoven; spores echin- о ee СТІП COM ИН. и гот Ç Rosemblinz H. spongiosus but many hyphae have the wall minutely spin lose or rough; known from New Hampshire and Massachusetts . 4.4... л сл el 58 вор &. Eh л * * e o9 e e во» s q өө ө» эз» ө ө ө э ө ө ө ө е ө э ө ө э э ө 4 n ө on ө ө ө gon ө ө ө n n ..4.4.ш.%...... Ka Ka > POR TOP Y Ku EE ENEE 8. H. spiniferus 10. Kéiere еи separable, 200-4004 thick; hyphae бы walled, loosely , 215-4u in diameter, with some rope-like strands ext 8 go ана по Қау астии color change — in se tions by KHO Boltin: «rere rm EEN d oo KC le H. granulosis 10. Fruetifieation D adnate, the color destroyed and ab d by KHO solution wh h beeomes colored brownis h; hypha да x -walled, 5-би in ae. PPA eer Tee е кна е амеры olivascens 11. EE ра SN Zeie" ya 75u thick; hyphae ch celled, gular and dia n diameter; spores grayish olive wik the а EE purs from New Hampshire ........ 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 207 11. Fruetifieation felty-membranaceous, light mineral-gray, 400и thick, two- кі 59 н e 17. Ts = =з mM -1 p я Without eystidia ; fructification Бо, buff to cinnamon layered; hyphae 4ш in diameter; spores deep olive-buff to hyaline under the microscope, rough-walled or aculeate with very short rom on ground Майда леш Ж онооно H. epigaeus 12. рос рео e eer from substratum when moisten od ERN 12, Fruetifieation adnate, fawn-color, under side and margin ке: apas етене € walled, 244 зи in ere hyaline under t cope; known from Washington................ . Кы Я KHO s solution causes a a color senate when added to sections immersed i drop of water in 1 REGELEN, зо EE 14 . KHO e AN no noteworthy pico chang, so sen aa EU eres 15 14. A change of color to tations blue-green and sage-green is caused in the granules; fructification Chaetura-drab to fuscous, mo. ular, the margin much paler, brownish and floccose; r Еа what colored, 3-4и in йіатебег...................... botryoides 14. A change of color to sage- EM is eaused in ps 'iymenium; ES cation brownish olive, granular, the rgin aceous-taw ma phae Cote. colored, only nds аа iate 215 Min i E diameter, forming occasional rope-like strands next to substratu 18. Н EE ОРОНОТ А енші 14. Original colors are destroyed and the hyphae pete sage-green ; fructification olive-ocher at surface, with under margin Ain ish drab; pd rues aad in diameter, SÉ some pod. -like hyphal ands next to substratum............................ 19. H. bicolor 3 Fruetifieation between icti mns and Vandyke-brown (a ‘‘dark red") and ^ margin Isabella-color or melleus; hyphae colored, 5-би in di- ameter, with rope-like gegen next the substratum ......... . atroruber š Fruetifieation with upper side pres buff E €— color, the under side and m Se, за hyphae, 5-74 in diameter, run along the sub- SES m and give off suberect, Fi ил Greg branches 315 = би in іп diameter — по rope-like strands ........................ Н. fuligineus 5. анна drab-gray, the margin whitish; hyphae hyaline under the EEN Ee EE кен E A . H. cinerascens 16. жеар not Geier Ee i. e. not having clamp connections ..... B 16. Hyphae подове-верігіе........................................... With cystidia ; fructification pinkish buff, adnate ..... 25. H. зын Without cy stidia ; fructification becoming warm buff, thick, and firm like се portentosum ; hyphae ди in diameter, terminating in the hymenium in dichotomously DT antler-shape ed organs; basidio м hyaline or nearly so; even spores, ова like the маға abun dot bebweensthe hyph8še..,...... deer аре rene ¿Sali zm Н. thelephoroides 4 о m ae 3 to substratum; spores with a sli Se tinge of buff in collection on slide but hyaline under the microscope, echinulate, the body 5-6x4-4% et cum SS ARMIN Sr T RIES MOREE Se ca IE ROR S pite PP я — — POR EN yellow to deep “colonial buff ; hypha for e 3-4u in dia not ing rope-like strands; spores con- А. жәні "e? sometimes. Fe as: the microscope, eehinulate, the EELER EE 28. H. ec hinosporus 18. Fruetifeation between olive-buff and — olive-buff ; т" color Wm ei $ or thé mieros6opëa, s ..... 29. oem DH. fibrillosus 18. Fruetifieation Comp? ge w to drab and ‘Fuscous, the ma d whitish or yellowish, flaxy- Поне, radiating; spores white in collection oo on slide, minutely echinulate with short, bien E^ body E TEE, i Vane 1. Hypochnus ferrugineus Pers. ex Fries, Obs. Myc. 2:280. 1818 and 1824; Karsten, Finska Vetenskaps-Soc. Bidrag Natur och Folk 37:162. 1882; Finl. Basidsv. 440. 1889; [VoL. 3 208 ANNALS OF THE MISSOURI BOTANICAL GARDEN Saec. Syll. Fung. 6:660. 1888; Bresadola, (Hym. Hung. Kmet.), I. R. Ассай. Agiati Atti III. 3:114. 1897. Corticium (Tomentella) ferrugineum Persoon, Obs. Мус. 2:18. 1799. — Thelephora ferruginea Persoon, Syn. Fung. 2:578. 1801; Myc. Eur. 1:141. 1822; Fries, Elenchus Fung. 1:198. 1828; Epicr. 543. 1838. — Corticium ferrugineum sub- genus Hypochnus Fries, Hym. Eur. 661. 1874. — Hypochnus ferruginosus (Fr.) Patouillard, Tab. Anal. Fung. 17. f. 26 1883. — Tomentella ferruginea Pers. ex Schroeter, Krypt.-F1. Schlesien 3:419. ; Illustrations: Patouillard, Tab. Anal. Fung. f. 26. Fructification effused, adnate, often suborbicular, thin, dry, tomentose, hypochnoid, drying Sudan-brown; structure in section about 3005 thick, composed of loosely interwoven, even-walled hyphae 414-5y in diameter, nodose-septate, concolorous through the whole fructification with the hymenium; no Н. e eystidia; basidia 4-spored; spores subglobose, Hypha, spore concolorous with the fructification, echinulate, gen body of spore about 7-8а in diameter. Fruetifieations 2-4 em. in diameter or 3-6 em. long, about 2-9 em. broad. Under side decaying limbs and logs of various frondose species. Canada and New Brunswick to Georgia and west- ward to Michigan. July to October. Occasional. This species is well marked by its very constant color, соттоп to both hyphae and spores, and its occurrence in ad- nate, small, and very thin, hypochnoid areas of the form and dimensions given. American collections agree closely in above respects with the European specimens received from Bresadola which he has noted as surely H. ferrugineus. Specimens examined :! Sweden: Femsjo, L. Romell, 225, 227. Austria-Hungary: Trentino, G. Bresadola; Tatra Magma, "With regard to the citation of b engen all except those of ‘‘ Exsiceati’’ are in Burt Herbarium, which are cited without explicit EE to place in other herbaria. For example, the ge imens cited “Sweden: Femsjó, L. Romell, 225, 227,” are in Burt Herbarium. The data given is that iio А with the 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 209 Lóese, V. Greschik, comm, by G. Bresadola. New Brunswick: Campobello, W. G. Farlow. New Hampshire: Chocorua, W. G. Farlow. Massachusetts: Belmont Spring, C. Bullard, comm. by W. G. Farlow; Sharon, A. P. D. Piguet, comm. by W. G. Farlow. New York: Alcove, C. L. Shear, 1316, in part; East Galway, E. A. Burt, two collections, Georgia: Tallulah Falls, A. B. Seymour, comm. by W. G. Farlow (in Mo. Bot. Gard. Herb., 43911). Wisconsin: Blue Mounds, Е. T. € S. A. Harper, 876. 2. H. rubiginosus Bresadola, (Hym. Hung. Kmet.), I. Е. Ассад. Agiati Atti ПІ. 3:114. 1897. Zygodesmus rubiginosus Peck, N. Y. State Mus. Rept. 30:58. 1879. — Tomentella rubiginosa (Bres.) R. Maire, Ann. Mye. 4:335. 1906. Type: in Bresadola Herb.; probably a portion in Burt Fruetifieations effused, membranaceous, somewhat separa- ble from the substratum, dry, tomentose, drying Brussels- brown; hymenium even or granular; structure in section about 200—300, thick, with all the hy- “4 phae bright-colored and giving their color to EE the fructification, about Зи in diameter, nodose- с septate, thin-walled, lax, loosely imterwoven towards the hymenium, longitudinally arranged тр. pubiginosus. next to Ше substratum, and occasionally con- Hyphal strand, solidated there in rope-like, branching strands “9 дама up to 15» in diameter; no eystidia; spores concolorous with the fructification or more intensely colored, subglobose-angu- lar, aculeate, body about 6-7» in diameter, or 7-8 6p. Fructifications about 115-3 em. long, 1-2 em. broad. On decaying leaves and decaying wood. Canada, New York, Louisiana, and British Columbia. October. Rare. specimens and may identify duplicates in another herbarium. The location of all specimens in herbaria other than my own is read E giving in paren- thesis the name of the herbarium Ae ded by * Fo мар» Ge EE cited ‘‘ Georgia: пра Falls, A. В. Sea 22 eomm. by ща w (in Bot. Gard. Herb., 43911),’’ is in Missouri Botanieal Garden А. дала but sc? in Burt ыы dis [VoL. 3 210 ANNALS OF THE MISSOURI BOTANICAL GARDEN H. rubiginosus is very similar in color throughout to H. ferrugineus but differs in being membranaceous, in having spores aculeate rather than spinulose, and in having some hyphae parallel with substratum and occasionally forming rope-like strands. These strands are not mentioned by Bresadola in his description, but they are present in prepara- tions from the specimen received from him and also in those from the few American collections referable to this species. Specimens examined: Hungary: on leaves of Juniperus and Quercus, Oct., 1888, Amet, comm. by G. Bresadola, apparently part of type. Canada: Lower St. Lawrence Valley, J. Macoun, 77. New York: Greenbush, C. H. Peck, type of Zygodesmus rubiginosus (in Coll. N. Y. State); Aleove, C. L. Shear, 1329; Syracuse, L. M. Underwood, 36, 41 (both in Coll, N. Y. State). Louisiana: St. Martinville, 4. B. Langlois, ct. British Columbia: Sidney, J. Macoun, 80, in part (in Mo. Bot. Gard. Herb., 8935). 3. H. subferrugineus Burt, n. sp. Type: in Burt Herb. Fructification effused, dry, membranaceous, separable from the substratum as a thin membrane, tomentose, drying Sudan- brown, with surface often granular in the center; structure in section 300—400, thick, CH composed of (1) a few dark-colored, nodose- EM _ septate hyphae 5-64 in diameter, running parallel with the substratum, loosely inter- Bee woven or sometimes im rope-like strands e и, which give off (2) suberect, bright-colored, . S gineus. ` 4 Hypha, врогехб40. interwoven branches, concolorous with the hymenium, bearing the basidia; basidia 4- spored; spores concolorous with the hymenium, subglobose, echinulate, with spore body 7-9 x 6-8; some color is dissolved from the sections when they are treated with KHO solution. Fructifications 2-5 em. long, about 2-3 em. broad. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 211 Under side of decaying limbs and logs of both coniferous and frondose species. Canada and New England to Michigan, and in British Columbia; also in Sweden. August to Octo- ber. Occasional. This species has the same color externally as H. ferrugi- neus, from which it differs in being more compact, so that it is membranaceous and may be cautiously peeled up from the substratum. Dried specimens often have their central portion cracked and curled away from the substratum, while H. ferrugineus is adnate. Furthermore, H. subferrugineus has hyphae next to the substratum dark-colored and ar- ranged longitudinally along the surface of the substratum, which is not the ease in H. ferrugineus. Specimens examined: Exsiecati: Ellis, N. Am. Fungi, 421, under the name Zygo- desmus rubiginosus. Sweden: Femsjo, L. Romell, 233. Canada: definite locality not stated, J. Macoun, 11; St. Lawrence Valley, J. Macoun, 20. New Hampshire: Chocorua, W. G. Farlow, 1, 3, a collection dated Sept., 1903, and a collection dated 1915 — the last (in Mo. Bot. Gard. Herb.). Vermont: Middlebury, E. A. Burt, two collections. New York: Sylvan Beach, Oneida Co., H. D. House (in N. Y. State Mus. Herb. and in Mo. Bot. Gard. Herb., 5893). New Jersey: Newfield, J. B. Ellis, in Ellis, N. Am. Fungi, 421. Michigan: Ann Arbor, 4. H. W. Povah, 4 (in Mo. Bot. Gard. Herb., 11774). British Columbia: Sidney, J. Macown, 26, in part (in Mo. Bot. Gard. Herb., 8933). 4, H. canadensis Burt, n. sp. Type: in Burt Herb. Fructifications small, effused, membranaceous, easily sep- arable from the substratum, dry, tomentose, drying between Brussels-brown and hazel, the margin very thin, fibrous; [Vor. 3 212 ANNALS OF THE MISSOURI BOTANICAL GARDEN hymenium even or granular; in structure 400-500,» thick, com- posed (1) next to the substratum of a few dark-colored, longitudinally arranged, nodose-septate hyphae 4-414» in diameter, and (2) towards Ше hymen- ium of pale, thin and even-walled hyphae about 22—34 in diameter, suberect, very loosely inter- woven, which arise as lateral branches from the dark basal hyphae and bear База and eystidia; eystidia septate, cylindric, obtuse, even-walled, Saccardo’s umber in color under the microscope, 4175-ӛр in diameter, emerging up to 80-1004; basi- dia 4-spored with the spores on slender sterig- mata about Dn long; spores Saceardo's umber un- der the microscope, globose, tuberculate, spore body 6-7» in diameter. Fructification usually 1-2 em. long, 15-1 em. + Fig. В broad, one specimen 4 cm. long. . eanaden- ° i a Cystid. On wood and bark of conifers decaying on the 5” forest floor. Canada апа New Hampshire to Ida- ho and British Columbia. August to November. Н. canadensis is a little darker in color than H. ferrugineus and is smaller and less conspicuous in the few collections which have been made. It differs from our other rust- colored species of Hypochnus in having eystidia. It is re- lated to the European Hypochnus ferruginosus (v. Hohn. & Litsch.) Burt, n. comb., = Tomentellina ferruginosa v. Hohn. & Litsch, by the colored, eylindrie eystidia, but the cystidia of our species are shorter and its hyphae finer, darker, and nodose-septate next to the substratum. Specimens examined: Canada: locality not stated, J. Macoun, 11. Quebec: Ironsides, J. Macoun, 277b. New Hampshire: Chocorua, W. G. Farlow, 2, and c4 (the latter in Mo. Bot. Gard. Herb., 44039). Vermont: Middlebury, E. A. Burt, type. Michigan: Ann Arbor, C. H. Kaufman, 36. Idaho: Priest River, J. R. Weir, 1. British Columbia: Kootenai Mountains, near Salmo, J. R. Weir, 504 (in Mo. Bot. Gard. Herb.). 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 213 5. H. umbrinus (Fries) Burt, n. comb. Thelephora umbrina Fries, Elenchus Fung. 1:199. 1828, but not T. wnbrina Alb. & Schw. Consp. Fung, 281. 1805. — Corticium umbrinum Fries, Hym. Eur. 658. 1874. — Thele- phora biennis Fries, Hym. Eur. 636. 1874, but not T. biennis Fries, Syst. Мус. 1:449. 1821. — T. arachnoidea Berk. 6 Broome, Linn. Soc. Bot. Jour. 14:64. 1873, but not T. arach- noidea as understood by Bresadola, Ann. Mye. 1:108. 1903. — Hypochnus tristis Karsten, Soc. pro Fauna et Flora Fennica Meddel. 9:71. 1883; Bresadola, Ann. Mye. 1:107. 1903. — Hypochnopsis fuscata Karsten, Finl. Basidsv. 443. 1889. — Hypochnus fuscatus Karsten in Басс. Syll Fung. 9:244. 1891. — Tomentella tristis (Karst.) v. Норте! & Litschauer, K. Akad. Wiss. Wien Sitzungsber, 115:1572. 1906. — Hypoch- nus sitnensis Bresadola, I. R. Ассай. Agiati Atti III. 3:115. 1897. Type: in Herb. Fries, and an authentic specimen from Fries in Kew Herb. Fructification effused, soft, separable, with the hymenial surface compact and membranaceous, varying from drab to fuscous and Chaetura-drab, underneath vil- lose; structure in section 400-600» thick, with [ү у some hyphae running along the substratum | y and ascending so as to form a loosely ar- Wy ranged layer near the substratum and then branching repeatedly to form a compact hy- & menium; hyphae concolorous with the fruc- ` tification, thick-walled, not nodose-septate, not Fig. 5 rough-walled, 4-би in diameter; basidia with H. umbrinus. 4 sterigmata; spores concolorous, globose or Ез тиер subglobose, aculeate or coarsely tuberculate, 6-7» in diameter or 6-8 X 415-7»; KHO solution dissolves some pigment from the sections and becomes dark-colored in their vicinity. Fructifications 6-10 em. long, 3-5 em. broad. On rotting coniferous and frondose wood. New England to British Columbia. September to October. Common and cosmopolitan. [Vor. 3 214 ANNALS OF THE MISSOURI BOTANICAL GARDEN Hypochnus wmbrinus (Fr.) is noteworthy among the dark species by its hyphae not being nodose-septate, i. e., not hav- ing clamp connections. Its tuberculate or aculeate spores and compact hymenium afford additional distinctive characters. Thelephora umbrina Alb. & Schw. is regarded now by European botanists as a Coniophora, of which I have a speci- men from Bresadola; what Fries understood by T. wmbrina is exaetly shown by an authentie specimen in Kew Herba- rium. This specimen is a true Hypochnus in fine condition, dark-colored, with compact hymenium separated from the substratum by a thick layer of loosely arranged, suberect, thick-walled, colored hyphae, which do not have clamp con- nections. Т. biennis, as used by Fries in 1821, is a deserip- tion of the illustration in Bulliard’s ‘Herb. de la France’ 2:986. pl. 436. f. 2, Fries stated that he had seen no speci- mens at that time. In ‘Hymenomycetes Europaei,’ published in 1874, he changed the description of T. biennis materially to adapt it to living specimens which he had seen. The re- supinate specimen of this later period in Herb. Fries is not distinct from Hypochnus umbrinus. Authentic specimens of H. tristis and Hypochnopsis fuscata received from Karsten, and of Hypochnus sitnensis from Bresadola are the same species as already pointed out by Bresadola;' still earlier, Romell stated in letters his belief that H. tristis is a synonym of H. wmbrinus. My studies lead to the same conclusion. The type specimen of Thelephora arachnoidea Berk. & Broome agrees closely with the Friesian specimen of H. wm- brinus. Bresadola? has deseribed hyphae of T. arachnoidea s **punetato-seabrae vel tunica granoso-aculeolata primitus induetae, usque ad ди crassae,’’ but in my preparation of the type of T. arachnoidea the walls of the hyphae are even and not more than 4%» in diameter.’ Ann, Myc. 1:107. 1903. 108. ?[bid., p. "In bg same connection оа е. Thelephora EN Ell. & Ev. а synon of T. arachnoidea, and he n follo SE s by von Hóhnel. My preparations of the t type of T. ло wie n N. that this species is not a basidiomyeete, CH that its ы ате nodose-septate. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 215 Specimens examined: Sweden: Smolandia, from E. Fries (in Kew Herb.); Femsjó, L. Romell, 234, 235, and E. A. Burt; Stockholm, L. Romell, 229—232. Finland: Mustiala, P. Karsten, authentie specimen of H. tristis; Messuby, P. Karsten, authentic specimen of Hy- pochnopsis fuscata. Hungary: A. Kmet, comm. by G. Bresadola, authentic speci- men of Hypochnus sitnensis. Ceylon: Habgalla, No. 539, Feb., 1868, the type of Thele- phora arachnoidea Berk. & Broome (in Kew Herb.). Canada: J. Macoun, 64. Ontario: Harraby, E. T. € S. A. Harper, 593, New Hampshire: Chocorua, W. G. Farlow, 9, 13, 14, 15, 23. Vermont: Middlebury, E. A. Burt. Massachusetts: Sharon, A. P. D. Piguet, comm. by W. G. Farlow. New York: Lake Placid, C. H. Peck; Floodwood, E. A. Burt. Wisconsin: Blue Mounds, E. T. Ф S. A. Harper, 860. British Columbia: Kootenai Mountains, near Salmo, J. R. Weir, 441, 487 (in Mo. Bot. Gard. Herb., 8227, and 20225 respectively). 6. H. fuscus Pers. ex Fries, Obs. Mye. 2:280. 1818 and 1824; Karsten, Finska Vetenskaps-Soc. Bidrag Natur och Folk 37:163. 1882. Corticium fuscum Persoon, Obs. Мус. 1:38. 1796; Fries, Hym. Eur. 651. 1874. — Thelephora fusca Fries, Syst. Myc. 1:451. 1821. — T'helephora vinosa Persoon, Syn. Fung. 2:578. 1801. — T'omentella fusca (Pers.) Schroeter, Krypt.-Fl. Schle- sien 3:419. 1888. Type: existence of an authentie specimen unknown to me. Fructification effused, membranaceous, separa- ble, einnamon-drab, darkening to Benzo-brown and Natal-brown; structure in section 200-350, thick, with a few hyphae running along the substratum ÉD and ascending and branching or giving off suberect, ^ spores | loosely interwoven branches; hyphae concolorous ` X940. with the fructification but rather pale under the microscope, [Vor. 3 216 ANNALS OF THE MISSOURI BOTANICAL GARDEN nodose-septate, 4-6 in diameter, sometimes collapsed; ba- sidia with 4 sterigmata; spores darker than the hyphae, subglobose, sometimes flattened on one side, the spore body 6-7» in diameter and short-aculeate in European and occa- sional American specimens, but more commonly 6-8 X ба and echinulate in American specimens. Fructifications 2-10 em. long, 1-2 em. broad. On rotten coniferous and frondose wood of several species. Canada and New Brunswick to New Jersey and in Montana. July to October. In the color of H. fuscus, there is a perceptible vinaceous component by which the species may be approximately rec- ognized at sight. Confirmatory characters are the separable fruetifieation and microscopical details of sections. The spores of most American specimens have slenderer and longer spines than those of European collections. H. fuscus is presented here as understood by Bresadola. Specimens examined: Sweden: Stockholm, L. Romell, 224. Hungary: A. Kmet, comm. by G. Bresadola. Canada: locality not given, J. Macoun, 14; Ottawa, J. Ma- coun, 28. New Brunswick: Campobello, W. G. Farlow, 4. Massachusetts: Magnolia, W. G. Farlow, two collections. New York: Albany, H. D. House & Jos. Rubinger (in Mo. Bot. Gard. Herb., 8736); East Galway, E. A. Burt; Pots- dam, J. B. Ellis (in Farlow Herb.). New Jersey: Newfield, J. b. Ellis (in N. Y. Bot, Gard. Herb., under the name Thelephora floridana). Montana: Missoula, J. R. Weir, 400 (in Mo. Bot. Gard. Herb., 22161). 7. H. spongiosus (Schw.) Burt, n. comb. Thelephora spongiosa Schweinitz, Naturforsch. Ges. Leip- zig Sehrift, 1:109. 1822; Am. Phil. Soc. Trans. N. S. 4:168. 1834; Fries, Elenehus Fung. 1:193. 1828; Sace. Syll. Fung. 6:545. 1888. — Hypochnus obscuratus Karsten, Hedwigia 35:46. 1896; басе. Syll. Fung. 14:226. 1900. 1916] | BURT—THELEPHORACEAE OF NORTH AMERICA VI 217 Type: in Herb. Schweinitz. Fructification effused, soft, felty-membranaceous, separa- ble, in color varying from Saccardo’s umber to bister, rarely fuscous, the margin thinning out and barely determinate; in structure 200-1200, thick, with hyphae concolorous with the fructifica- tion, thick-walled, even, loosely interwoven, branching at a wide angle, abundantly no- dose-septate, 414-5u in diameter or rarely би; basidia with 4 sterigmata; spores con- Fig. 7 colorous, globose, or subglobose and flattened Eo Sige on one side, echinulate, about 6 in diameter, x 640. or 6-9 X 6-7и. Fructifications 4-10 cm., and more, long, 2-5 em. broad. On rotten wood and bark of both frondose and coniferous species. Canada to North Carolina and westward to Mon- tana, and in Bahama Islands. July to November. Probably common. H. spongiosus belongs in the group with H. fuscus, H. um- brinus, and H. spiniferus. The absence of a vinaceous com- ponent in its color is a useful character for separation at a glance from H. fuscus. If the surface of H. spongiosus is viewed with a lens, the component fibers are seen running in all directions, as in felt or blotting paper. H. wmbrinus has its hyphae lacking clamp connections, i. e., not nodose- septate, and its basidia form a compact hymenium. H. spin- iferus differs by having its hyphae spiny. Specimens examined: Finland: Mustiala, P. A. Karsten, authentic specimen of Hypochnus obscuratus. Canada: Quebec, Ironsides, J. Macoun, 255. New Hampshire: Chocorua, W. G. Farlow, 14. Vermont: Middlebury, E. A. Burt, three collections; Lake Dunmore, E. A. Burt. New York: Albany, H. D. House (in N. Y. State Mus. Herb. and in Mo. Bot. Gard. Herb., 15833). North Carolina: Schweinitz, type (in Herb. Schweinitz). Indiana: Miller, E. T'. & S. A. Harper, 758. [Vor. 3 218 ANNALS OF THE MISSOURI BOTANICAL GARDEN Wisconsin: Lake Geneva, E. T. € S. A. Harper, 950. Montana: Evaro, J. R. Weir, 436, 438 (in Mo. Dot. Gard. Herb. 19515 and 19597 respectively). Bahama Islands: 4. E. Wight (in Farlow Herb.). 8. H. spiniferus Burt, n. sp. Type: in Farlow Herb. and in Burt Herb. Fructification effused, membranaceous, separable, tomen- tose, varying from sepia to fuscous; in structure about 1000. thick, with the hyphae loosely interwoven, nodose-septate, thick-walled, concolorous with the fructifieation but darker near the substratum and spinulose, the paler hy- phae rough-walled or even, body of largest I hyphae 4—5z# in diameter, the spines about piniferus la long, colored like the dark wall; basidia with 4 sterigmata; spores concolorous, globose, sometimes flattened on one side, echinulate, the body 6-8, in diameter, or 6 X 414-6n. Fructifications about 5 em. long, 3 em. broad. On rotten wood. New Hampshire and Massachusetts. Au- gust. Rare. H. spiniferus is so similar to H. spongiosus in habit and coloration that it can be separated from the latter only by the distinctly spiny-walled and rough-walled hyphae of the former species. This character is as marked as in the capil- litium of some Myxomycetes. The New Hampshire collec- tions which I have included under H. spiniferus have rough- walled hyphae and no spines. Specimens examined: New Hampshire: Chocorua, W. G. Farlow, 11, and an un- numbered specimen collected in 1904. Massachusetts: Magnolia, W. G. Farlow, type. . В u Hypha, sporex 640. 9. H. granulosus (Peck) Burt, n. comb. Grandinia tabacina Cooke & Ellis, Grevillea 9:103. March, 1881, but not Hypochnus tabacinus Bresadola. — Zygodesmus granulosus Peck, Bot. Gaz. 6:277. 1881. — Hypochnus elae- odes Bresadola, I. R. Ассай. Agiati III. 3:115. 1897. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 219 Type: in Coll. N. Y. State. Fructification effused, thin, membranaceous, separable from the substratum, granular, sepia, the margin somewhat radiate, concolorous or nearly so; in struc- ture 200—400, thick, composed of very loosely e interwoven, thin-walled, occasionally nodose- E septate, hyphae 214-4» in diameter, yellow- e са ish under the microscope, forming near the substratum some rope-like mycelial strands H. granulosus. up to 15и in diameter; spores concolorous Spore, hyphal ; strand x 640. with the hyphae, angular-subglobose, acule- ate, the body about 64 in diameter; KHO solution produces no noteworthy color change in sections. Fructifications 2-4 em. long, 1-2 em. broad. On rotten bark and wood of frondose species. Massachu- setts to New Jersey and Ohio. September to November. Rare. H. granulosus is very closely related to H. coriarius and is distinguished from it by uniform color of the whole sur- face, while H. coriarius has the margin ochraceous-tawny. The lack of noteworthy color change by KHO solution is the only additional feature of difference for separating H. gran- ulosus from H. coriarius. The specific name tabacina of Cooke and Ellis has priority, but is not now available be- cause Bresadola has already used the name Hypochnus ta- bacinus for a valid species. Specimens examined: Exsiceati: Ellis, N. Am. Fungi, 421, under the name Zygo- desmus chlorochaites. Hungary: A. Kmet, authentic specimen of H. elaeodes from Bresadola, probably a portion of the type. Massachusetts: Newton, W. G. Farlow; Mt. Tom, H. W. Harkness, type (in Coll. N. Y. State). New York: Albany, H. D. House & J. Rubinger (in Mo. Bot. Gard. Herb, 8733); Karner, H. D. House (in Mo. Bot. Gard. Herb., 44731); Alcove, С. L. Shear, 1316, in part. [Vor. 3 220 ANNALS OF THE MISSOURI BOTANICAL GARDEN New Jersey: Newfield, J. B. Ellis, in Ellis, N. Am. Fungi, 421, and also the cotype of Grandinia tabacina (in N. Y. Bot. Gard. Herb.). Ohio: A, P. Morgan, 525 (in N. Y. Bot. Gard. Herb., under the manuscript name Odontia olivacea). 10. H. olivascens (Berk. & Curtis) Burt, n. comb. Zygodesmus olivascens Berk. & Curtis, Grevillea 3:145. 1875. Type: type and cotype in Kew Herb. and in Curtis Herb. Fruetifieation effused, thin, not separable, tomentose, ci- trine, yellowish citrine or buffy citrine, the margin thinning out; КНО solution dissolves some of the color 435 upon coming іп contact with the sections and becomes somewhat brownish in their vicinity; Fig. 10 in structure 150—200, thick, with now and then Н. olivascens. д hypha running along the substratum and Spore x 640. Lë А sending out suberect branches which branch re- peatedly, become loosely interwoven, and are somewhat clus- tered; basal hyphae slightly colored, nodose-septate, thin- walled, 5-64 in diameter; basidia with 4 sterigmata; spores subglobose, concolorous with the basal hyphae, aculeate-echin- ulate, the body about би in diameter or 514-714 X 510-7. Fruetifieations sometimes in little patches 1-2 em. long, 115-1 em. broad, sometimes growing more or less interrupt- edly over areas up to 15 em. long, 3 em. broad. On very rotten wood and on bark of fallen branches of both coniferous and frondose species. New Brunswick to South Carolina. September to November. Probably com- mon. H. olivascens is readily distinguished from other species of Hypochnus by its conspicuous citrine color of some kind (flavovirens of Saecardo's ‘Chromotaxia’) which has been retained well by the original collection for more than sixty years. From the description, Tomentella flavovirens v. Hohn. & Litsch. is but slightly, if at all, different from H. olivascens. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 225 Specimens examined: Exsiceati: Ellis, N. Am. Fungi, 422, under the name Zygo- desmus olivascens. New Brunswick: Campobello, W. G. Farlow, 5. New Hampshire: Chocorua, W. G. Farlow, 5, 6, 18. Vermont: Weybridge, E. A. Burt. Massachusetts: Magnolia, W. G. Farlow; Hyde Park, C. Bullard, comm. by W. G. Farlow; Sharon, A. P. D. Piguet (in Farlow Herb.) ; Stony Brook, G. В. Lyman, 167; Wil- liamstown, W. G. Farlow, 7. New York: North Greenbush, H. D. House, two collections (in N. Y. State Mus. Herb. and in Mo. Bot. Gard. Herb., 14852, 20191); Karner, H. D. House (in N. Y. State Mus. Herb. and in Mo. Bot. Gard. Herb., 44719); Ithaca, C. Thom, Cornell Univ. Herb., 13582. New Jersey: Newfield, J. B. Ellis, in Ellis, N. Am. Fungi, 422. Pennsylvania: Kittanning, D. R. Sumstine. Maryland: Takoma Park, C. L. Shear, 1064, 1082, 1092. South Carolina: Society Hill, M. A. Curtis, eotype (in Curtis Herb., 5204). 11. H. pilosus Burt, n. sp. Type: in Burt Herb. Fructification effused, byssoid, membrana- ceous, separable from substratum, dry, tomen- tose, drying Sayal-brown, the margin slightly 4 paler, thin, narrow; hymenium even in places, somewhat granular and pitted elsewhere; struc- ture in section 200-800, thick, composed of hy- phae about 4-414» in diameter, branching at e right angles, of the same color as the fructifica- tion, nodose-septate, rather rigid, very loosely interwoven, somewhat longitudinally interwov- en next to the substratum; cystidia septate, Fig. 11 sometimes granular incrusted, with the emer- gent portion colorless, thin-walled, cylindric, 514-64 in diameter, emerging 40-90и, tips ob- tuse or clavate; spores 4 to a basidium, slightly darker than H. pilosus. Spore, cystidi- umx 640. [Vor. 3 222 ANNALS OF THE MISSOURI BOTANICAL GARDEN the hyphae, subglobose-angular, aculeate, the spore body 7-9 ж би. Fructification 8 em. long, 2-3 em. broad — broken off at one On bark of decaying Quercus alba, Lake Geneva, Wiscon- sin, July. This fungus suggests Coniophora arida and C. puteana by its umber color and broadly effused fructifications, but it is a true Hypochnus, which is readily distinguished from other species of this genus by its color, hair-like cystidia, and the spores. Specimens examined: Wisconsin: Lake Geneva, E. T. € S. A. Harper, 877. 12. H. isabellinus Fries, Obs. Myc. 2:281. pl. 6. f. 3. 1818 and 1824; басе. Syll. Fung. 6:657. 1888; Bresadola, Ann. Mye. 1:106. 1903. Corticium isabellinum (in section Hypochnus) Fries, Hym. Eur. 660. 1874. — H. argillaceus Karsten, Soc. pro Fauna et Flora Fennica Meddel. 6:13, 1881; Sace. Syll. Fung. 6: 661. 1888. Type: there is a specimen from Е. P. Fries in Curtis Herb. Fruetifieation effused, tomentose, thin, adnate, varying from deep olive-buff to dark olive-buff, the margin thinner, concolorous; in structure 60—200y, rarely 300и, thick, with a few hy- рһае 8-10и, or more, in diameter, ad P d running along the substratum and sending out suberect, loosely inter- woven branches; hyphae concolor- Fig. 12 ous with the fructification, branch- H. isabellinus. ing at right angles, thick-walled, Spore, hypha x640. not nodose-septate; basidia with 4 sterigmata; spores concolorous, globose, echinulate, the spore body 7-9& in diameter. Fruetifieation 5-10 em. long, 144-3 em. broad, and probably larger. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 22d On rotten wood and bark of both coniferous and frondose species. Canada to Florida, in Wisconsin and in Jamaica. May to January. Probably common. H. isabellinus is a little thinner and a little paler than H. pannosus, and not separable from the substratum in the collections which I have studied. It is best distinguished from the latter species by the larger hyphae of H. isabellinus and lack of clamp connections. Specimens examined: Exsiecati: Ravenel, Fungi Am., 57b, under the name Zygo- desmus pannosus; Thiimen, Мус, Univ., 2275, under the name Zygodesmus pannosus. Sweden: Upsala, Halmbyboda, from E. P. Fries (in Curtis Herb.) ; Stockholm, L. Romell, 219—222; Femsjö, L. Romell, 223, and H Fries (in Herb. Fries under the manuscript name Hypochnus leprosus). Canada: Rockcliffe Park, J. Macoun, 144; St. Lawrence Val- ley, J. Macoun, 2. New Hampshire: Chocorua, W. G. Farlow, two collections. New Jersey: Newfield, J. B. Ellis, in Thümen, Mee, Univ., 2275. Florida: Gainesville, H. W. Ravenel, in Ravenel, Fungi Am., 91b. Wisconsin: New London, E. T. € S. A. Harper, 949; Stevens Point, C. J. Humphrey, 1948 (in Mo. Bot. Gard. Herb., 4748). Jamaica: Cinchona, W. A. ё Edna L. Murrill, N. Y. Bot. Gard., Fungi of Jamaica, 630. 13. H. pannosus (Berk. & Curtis) Burt, n. comb. Zygodesmus pannosus Berk. E Curtis, Grevillea 3:112. 1875. Type: cotype in Curtis Herb. Fructification effused, byssoid-membranaceous, separable when well developed, tomentose, varying in brown from Sac- cardo’s umber and snuff-brown to cinnamon-brown, the mar- gin concolorous and thinning out; in structure 120—350, thick, with an occasional hypha running along the substratum [VoL. 3 224 ANNALS OF THE MISSOURI BOTANICAL GARDEN but composed for the most part of suberect, branching, loosely interwoven, nodose-septate, thick- ы walled һурһае concolorous with the fruc- SEH Es tification, 4-64 in diameter; basidia with 4 sterigmata; spores concolorous with the fructification, subglobose, sometimes flat- tened on one side, echinulate, the body Fig. 13 . pannosus, 6-8 X 9-7, | Spore, hypha x640. Fruetifieation 3-6 em, long, 142-3 em. broad. On rotten wood and bark, usually of frondose species, and on the ground in woods. Canada to Louisiana; occurs in Europe also. September to December. Probably common. H. pannosus and H. isabellimus are species of brown color approaching clay-color, and of cottony surface, which cannot be distinguished from each other with certainty except by microscopic characters. Well-developed fructifications of H. pannosus are thicker than those of H. isabellinus but thin fruetifieations of the former are frequently collected. H. pannosus has nodose-septate hyphae 4—64 in diameter, while the hyphae of H. isabellinus are not nodose-septate and next to the substratum are 8-104, or more, in diameter, and occa- sionally 15a in diameter. KHO solution produces no note- worthy color change. The collection from Washington, re- ferred with doubt to this species, has the spores with body 6 X 41%и, aculeate with scattered, very short points. Specimens examined: Sweden: Stockholm, L. Romell, 225; Femsjo, L. Romell, 228. Canada: Quebec, Ironsides, J. Macoun, 277 a. New Hampshire: Chocorua, W. G. Farlow, 7, 8, and an unnumbered specimen; Shelburne, W. G. Farlow, 1. Vermont: Middlebury, E. A. Burt. Massachusetts: Magnolia, W. G. Farlow, c; Williamstown, W. G. Farlow, 5. South Carolina: Santee Canal, Ravenel, 1117, cotype (in Curtis Herb., 3007). Louisiana: St. Martinville, 4. В. Langlois, cs. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA VI 225 "Washington: Bingen, on Pinus ponderosa, W. N. Suksdorf, 860. 14. H. avellaneus Burt, n. sp. Type: in Burt Herb. Fruetifieation effused, soft, membranaceous, separable, upper side between cartridge-buff 25 and olive-buff and under side fuscous, the mar- gin narrow, radiate, colored like the upper sur- Дъ face or whitish; in structure 300-400и thick, with the hyphae snuff-brown under the micro- Fig. 14 scope, thick-walled, nodose-septate, rather com- н. avellaneus. pactly interwoven; basidia 4-spored; spores И три Жл concolorous with the hyphae, angular-subglo- bose, aculeate, the body 6-7% X би. Fructification 5 em. long, 1 em. broad. On wood of red fir in woods. Washington. October. This species is marked by the pale color (nearly avellaneus of Sacecardo’s ‘Chromotaxia’) of the upper surface and mar- gin and the fuscous subiculum. Specimens examined: Washington: Olympia, C. J. Humphrey, 6305, type. 15. H. sparsus Burt, n. sp. Type: in Farlow Herb. and in Burt Herb. Fruetifieation effused, very thin, byssoid, not forming a membrane, adnate, drab, the margin of the same color, indeterminate; in structure 60— 75 thick, with the hyphae hyaline under the mi- croscope, short-celled, irregular in form and di- Fig. 15 ameter, nodose-septate; basidia 4-spored; spores grayish olive under the microscope, echinulate, 6-7 X би; no noteworthy color change by KHO solution. Fructification 2-3 em. long, 1-2 em. broad. On bark of fallen frondose limbs. New Hampshire. x 50и. The terminal cells may often attain a length up to 600». According to Linossier and Roux (799), this organism does not grow in saliva. This peculiarity accounts for the fact that thrush occurs only in infants, more frequently during the first few months of life when the sali- vary secretion has been insufficiently established, and, in general, in all cases of infection accompanying a diminution of the secretion of saliva. On different media the fungus develops either by budding, like a yeast, or by the elongation and division of cells, as in Monilia. On carrot the mycelium is very well developed, whereas in nutrient liquids only yeast cells are present. Ас- cording to Vuillemin, the filamentous form is the normal method of vegetation, the yeast cells appearing in conditions of malnutrition. In sugar solutions at a temperature of 30-35°C., chlamydospores form at the end of the filaments. The asci were discovered by Vuillemin (298) in old cul- tures on beet. They appear as large, ovoid or elliptical asci, 4-5» in diameter, formed by a lateral or terminal bud- [Vor. 3 260 ANNALS OF THE Missourt BOTANICAL GARDEN ding of the mycelium or derived by the germination of chlamydospores. The aset contain four ascospores, formed in a manner similar to those of E. capsularis, slightly reni- form, 2.83.5 x 1.75-2 x 12-14, with a thick membrane. The germination of these ascospores has not been observed. Development takes place at a temperature of 20-39°С. on slightly acid, solid, or liquid media. In sugar solutions and fruit juices growth takes place slowly, with the formation of а flocculent deposit but with no film on the surface. The fungus coagulates milk after 20-30 days and ferments dex- trose slightly. Certain authors believe that there exist many varieties of E. albicans, only some of which have the func- tion of producing spores. Castellani (711) isolated from cases of bronchomycosis in Ceylon, twenty-two strains of Епфотусез which, in micro- scopic appearance and cultural characters, closely resembled E. albicans. Fourteen strains were identical and corre- sponded to E. tropicalis, whereas the other eight strains dif- fered from E. tropicalis, and from each other. The behavior of the different strains, especially toward sugar solutions, indicated that there are nine different species. Castellani believed that six of these species were parasitic, but he was not certain about the other three. It may be possible that a similar condition as to the plurality of species of Endomqyces affecting man also exists in the temperate zone. Before describing the better-known pathogenic yeast-like fungi, it may be well to consider first the development of the parasitic theory of cancer. This disease, more than any other human ailment, has been a fruitful field for the dis- covery of such forms as resemble yeasts. The one common characteristic of cancers is the power of cell proliferation, and the problem that many scientists to-day are undertaking is the causes underlying such proliferation. Many investigators take the view that cancer has some spe- cific and demonstrable cause. By the continued division of the carcinoma cells, masses of tissue are formed which grow out into lymph channels. Mechanically obstructing the normal activities of surround- 1916] Еміс--Үкавт Fuxar 261 ing tissues, or breaking through sueh tissues, they give off small groups of free cells which may be carried by the blood to various parts of the body, there to set up independent growths (metastases). With the local disturbances caused by such abnormal growths, many normal cells are killed, whereas the cancer cells themselves undergo hyperplasia and hypertrophy. The progress of cancer is accompanied by the degenerating of various kinds of cells, and these different structures are the things which have been interpreted by various investigators as x-bodies, amoebae, coccidia, pro- tozoa, or other organisms. Many of the structures thus interpreted as organisms are characterized by capsules which some investigators have in- terpreted as parts of an invading organism. Cell invasions are common in cancer tissue, but the capsules are only con- densations of the invaded protoplasm. Pianeze (296) ob- served similar bodies in the nuclei of cancer cells, and inter- preted both these and the cytoplasmic forms as colloidal degenerations of the chromatin and cytoplasm. Although these cell inclusions in human cancer cannot be considered as organisms, it does not follow that real organ- isms are not present. Later stages of the disease are par- ticularly suitable for secondary infection, and exposed sur- face lesions form a suitable medium for the growth of bac- teria, yeast-like organisms, and other fungi. San Felice ('95) made a series of inoculation experiments on animals with yeast-fungi obtained from various sources, principally from the juice of fruits. During the course of these experiments he found one species, T'orula neoformans, which, if inoculated into animals, produced the formation of neoplasm. The same author (’96) also discovered Cryptococ- cus lithogenes in a lymphatic ganglion of a cow that died as a result of a primary carcinoma of the liver. From this observa- tion he seemingly obtained the confirmation of his ideas on the pathogenic rôle of eryptococci in the formation of malignant tumors. His parasitic theory of cancer was based on the presence of cryptococci in tumors, the isolation of eryptococci [Vor. 3 262 ANNALS OF THE Missourr BOTANICAL GARDEN from diseased tissues, and the results of inoculation experi- ments on animals. The presence of cryptococci in tumors has been mentioned by a large number of authors, but the tumors thus parasitic do not all pertain to the type of malignant tumors. Binaghi (796) investigated fifty-three cases of epithelioma and iso- lated parasitic organisms in forty instances. The failure to obtain parasites in the remaining thirteen, according to this author, may be due to the fact that the part examined was either in an early stage of development or not infected. These organisms, identical with eryptococci in their mor- phological and physiological characters, were not found in other pathological or normal tissue. Consequently, they were considered as the specific cause of epithelioma. Maffucci and Sirleo (798) obtained ten or more cultures of yeasts from the thirty-nine tumors which they examined. Only one was pathogenie for guinea-pigs, and in these animals it produced fibrinous pneumonitis and abscesses under the skin or in the kidneys. The results indieated that a new formation of sarcoma tissue did not take place. КопсаП, Corselli, and Plimmer obtained cultures of fungi from malignant tumors, but only in a few cases were they pathogenic for animals. Leopold (700) reported having isolated pure cultures of сгур- іюсоссі from over 80 рег cent of the non-uleerated tumors investigated, the cultures having been obtained from the center of the diseased tissue. Loeb, Moore, and Fleisher (713) were unable to confirm the results of Leopold. Of the seventeen tumors examined, only one gave a culture of a yeast-like organism. In no case of human cancer has the causative significance of a microor- ganism so far been proved. Moreover, Busse (’03) was never able to obtain a single culture of Cryptococcus from non-ul- cerated tumors. No one has been able to demonstrate the development of tumors histologically comparable to cases of sarcoma, by the inoculation experiments on animals with cultures of Cryptococcus. By inoculation of C. tumefaciens in the white rat, Curtis obtained results in which ‘‘la tumeur etait iden- 1916] Emic—Yerast FUNGI 263 tique a celle de l'homme, c'est-a-dire constituée par une infiltration du parasite dans les mailles du tissue cellu- laires." "This, however, does not imply that Curtis obtained a sarcoma-like tumor. FUNGI IMPERFECTI CRYPTOCOCCUS The species of Cryptococcus here given were reported as having been obtained from tumors, but their róle in the produetion of these malformations has not been determined. C. Corsellii (Corselli & Frisco) Neveu-Lemaire, in Corselli & Frisco, Centralbl. f. Bakt, I. 18 :368-373. 1895. This species was isolated by Corselli and Frisco ('95) from a sarcoma of the mesenterie ganglia of a man. This fungus has dark cells, globose, and of variable dimensions. It is easily eultivated on gelatin, agar, and bouillon, neutral or alkaline. It can give rise to a slight fermentation and is pathogenie for guinea-pigs, dogs, and rabbits, by intraperito- neal inoculations. C. degenerans Vuillemin, in Roncali, Centralbl. f. Bakt. І. 18 :353-368. 1895. Roneali (295) observed this species in the ganglion of the armpit of a woman affected with cancer of the breast. In the cancer the cells are globose, rarely oval or reniform, isolated or in groups. In cultures the cells are globose or elliptical. On sugar nutrient liquids this fungus produces a film. On gelatin the colonies are irregular in outline and of a light yellow color. Gelatin is not liquefied and sac- charose is not fermented. It is pathogenic for the guinea- pig by intraperitoneal injections. The autopsy revealed an abscess of the mesenteric ganglia, which was produced by degeneration products; the normal cells were rarely found in these lesions. Cryptococcus of Gotti & Brazzola, in Guilliermond, Les Levures. 1913. This species was found by Gotti and Brazzola in a myxosarcoma of the nasal fossa of a horse. The cells are [Vor. 3 264 ANNALS оғ THE Міввотуві BOTANICAL GARDEN of variable dimensions, round or oval, with granular con- tents, enveloped by a membrane of double contour and a mucilaginous capsule that is often stratified. On gelatin or agar, the colonies are white with denticulate margins. Acid gelatin is liquefied. It is pathogenic for the guinea-pig but not for other animals. C. farciminosus Rivolta & Micellone, in Fermi & Aruch, Centralbl. f. Bakt. I. 17:593-600. 1895. This species has been considered as the parasite of “(African glanders." Тһе cells are globose or oval, some- times acuminate at the two poles. It grows with difficulty on all media. Fermi and Aruch have described the globules, which they considered ascospores, in the cells of this yeast. C. Gilchristi Vuillemin, in Gedoelst, Les Champ. Par. 1902. This fungus was obtained by Gilchrist in a case of scrofulo- dermatitis. It is only slightly pathogenic for animals. C. granulomatogenes (San Felice) Vuillemin, in San Felice, Zeitschr. f. Hyg. 44:364-396. 1903. This fungus was discovered by San Felice in the lung nodules of a hog. It is only slightly pathogenic for animals. C. hominis (Busse) Vuillemin, in Busse, Die Sprosspilze, in Kolle & Wasserman, Handbuch 1:631-700. 1903. This species was discovered by Busse (’03) in periosteal lesions of the tibia of a woman. Im situ the oval or globular cells are united in variable numbers in a substance of a homo- geneous aspect, constituting a sort of common capsule. This homogeneous substance is not present in cultures. The cells have a double-contoured membrane that becomes thicker with the age of the culture. It is easily cultivated on all media between 15 and 38°C. In liquid media it forms a deposit of yeast cells and a thin film on the surface. It does not liquefy gelatin. On potato the colonies rapidly unite to form a thick white layer. The fungus ferments dextrose and is pathogenic for rabbits, white mice, and dogs. C. linguae-pilosae (Raynaud & Lucet) Vuillemin, Lucet, in Archiv. d. Par. 4:262-287. 1901. 1916] Emic—Yerast Funct 265 This fungus was discovered by Raynaud and Lucet in the disease of ‘‘black-tongue.’’ Lucet (701) in his experimental studies demonstrated that this fungus did not reproduce the disease. According to Guéguen and Thaon, this yeast acts only when associated with Oospora lingualis. "There is, then, in these two fungi a sort of symbiotie relation. This organism has spherieal, ovoid, or elliptical cells, 4—17 X 6». It grows on most media and in liquids forms a white film after ten hours at 37°C. It ferments glucose and levulose, with the produetion of alcohol and earbon dioxide. Development is accompanied by the production of acid, but no liquefaction takes place on gelatin. It has a slight patho- genie action on certain animals and is without effect on guinea-pigs and rabbits by subcutaneous or intraperitoneal inoculations. C. lithogenes (San Felice) Vuillemin, in San Felice, Zeitschr. f. Hyg. 21:32-58, 394-420. 1895-96. San Felice isolated this parasite from the lymphatic gan- glia of a cow that died as a result of primary carcinoma of the liver, with extensions of the infection to the entire lym- phatie system. In the tissues the cells are more or less spherieal, of variable dimensions, enclosed by a refringent membrane. This species grows well on all media. On glu- cose bouillon it forms an abundant deposit of yeast cells and often a film on the surface. It does not liquefy gelatin. It forms white yeast-like colonies on agar, and on potato the colonies become dark brown in color. It is pathogenic for the guinea-pig, mouse, and sheep. C. niger (Maffucci & Sirleo) Vuillemin, іп Maffucci & Sirleo, Zeitschr. f. Hyg. 27:1-30. 1898. This species was discovered by Maffucci and Sirleo (798) in a pulmonary lesion of a guinea-pig inoculated with the liver of an embryo taken from a tubercular mother. It is pathogenic for animals only after a long time. The cultures sterilized by heat are toxic for guinea-pigs. C. Tokishigei Vuillemin, in Tokishige, Centralbl. f. Bakt. 1. 19:105-113. 1896. [VoL. 3 266 ANNALS OF THE Missourt BOTANICAL GARDEN This fungus is considered by Tokishige (’96) as produe- ing in Japan the disease of horses known as *'farcin." It is non-pathogenic for rabbits, guinea-pigs, and dogs, by sub- cutaneous inoculations. The pathological products of this fungus have more effect on animals than inoculations of the cultures. Inoculation of this organism in the horse pro- duces lesions after varying lengths of time, but without caus- ing the death of the animal. OOSPORA The longest-known fungous disease of the skin is favus, which previous to 1839 included a large number of different skin affections. When its contagious nature was discovered by Schonlein, investigators began searching for the relation of fungi to skin diseases. Heusinger in 1826, and Remak in 1837 (both cited by Virchow, 756) observed mould-like fila- ments in the seales of tinea. The latter author in 1847 suc- ceeded in growing cultures on apple, again producing the characteristic lesions, presumably by inoculating his arm with these cultures. Gruby (’43) independently discovered three different fungi associated with as many types of ring- worm. Two of these fungi were Oospora, and the other was Sporotrichum (Microsporon) Audouin. His description of these fungi was very good, yet no one at that time suspected that, under the name of Porrigo decalvans, he was describ- ing fungi which were the cause of ringworn. Very little attention was given to these discoveries until Sabouraud be- gan his investigations in 1892, which differentiated the many varieties of Oospora (Trichophyton) and Sporotrichum ( Mi- crosporon). | Oospora is often associated with mycosis of the lung and with Endomyces albicans in the mouth. Oospora pulmonalis brings about a degeneration on the surface of the trachea and walls of the bronchia, where white granules are found similar to those caused by Actinomyces. The alveolar struc- ture is filled with mycelium, and in certain places real ab- scesses are found constituted by degenerated tissue filled with mycelial debris. In lesions of the mouth, Roger, Bory, win 1916] Emic—Yrast FUNGI 267 and Sartory (’09) discovered Oospora buccalis associated with Endomyces albicans. In other diseases caused by Oos- pora, the fungus may occur as a saprophyte or as a para- site. These fungi are very closely related to each other and cannot always be differentiated sharply. Oospora of Trichophyton. Gruby (743) was the first to give a clear description of the Oospora parasite that causes epidemic and endemic diseases of the skin. The morphology of these fungi has been studied in the lesions which they produce and in cultures on artificial media. This fungus develops into filaments composed of short segments. The dimensions of the cells in the same filament are approximately equal. Their diameter remains invariably the same for the same species, but the length of the cells may vary 1 to 2и. The cells may be spherical or oval, the mycelial filament being moniliform and easily disso- ciated into its different elements. When the mycelial cells are cubical, the filaments are more or less extended and not easily dissociated. The classification of the varieties affecting man, according to the infection, falls into two groups. The endothrix group contains the varieties of human origin which develop in the hair between the cuticle cells and grow exclusively within the hair structure. The ectoendothrix group of parasites, probably of animal origin, develops in the hair and prolifer- ates in the follicle outside. More often the scheme of Sabouraud is used, in which are considered the mode of infection and principally the cultural characters of the isolated fungus when grown on ‘‘proof agar.” There are over thirty varieties of Oospora of Trich- ophyton, and their differences are chiefly cultural. Matruchot and Dassonville (’99), reasoning from analogies, place the parasites of Trichophyton in the Ascomycetes, in the family Gymnoascaceae. The asexual type of development in the Gymnoascaceae, according to these investigators, can serve to characterize these fungi with the same degree of precision as the complete forms. The demonstrated affinity [Vor. 8 268 ANNALS OF THE Missourt BOTANICAL GARDEN of Oospora tonsurans with Ctenomyces rests on the simi- larity of characters in the conidial stage of development. In old cultures of O. tonsurans may be found multicellular chlamydospores, spindle-spores, and serrate septate hyphae. This fungus, according to Matruchot and Dassonville, is an imperfect form of a species of Ctenomyces, still unknown, which has adopted a parasitic mode of development and con- sequently has lost the faculty of producing perithecia and ascospores. The serrate spiral hyphae, present in cultures of O. tonsurans, are considered as traces of asci formation. O. tonsurans (Malmsten) Saee. & Trav. in басс. Syll. Fung. 20:236. 1911. The mycelium of this fungus usually fills the entire hair without having passed through the cuticle. The filaments are more often simple, rarely dichotomously branched. The fungus is made up of squarish cells 4-би long, arranged in chains that follow the direction of the hair. O. porriginis (Mont. & Berk.) басс. Syll. Fung. 4:15. 1886. Oidium porriginis Robin, Veg. Par. 477-488. 1853. Gruby (743) demonstrated that the fungous parasite which he had independently found in Тале lesions, was the cause of the affection, and in the following year made successful inoculations with this fungus in the human skin and the skin of animals. In 1845 Remak separated the fungus from the genus Oidiwm to which it had been assigned, and created the genus Achorion, with the name Achorion Schonleini for this specific fungus. The characteristic lesion is a small yellow disc with a eup-like depression in the center. Both in color and in shape, it resembles a honeycomb, hence the name which comes from the Arabie Sahafts, meaning honeycomb. In the middle ages the disease was called tinea, which name is still retained. Matruchot and Dassonville (’99) also con- sider O. porriginis as belonging to the family Gymnoascaceae, but this classification cannot be regarded seriously. SPOROTRICHUM S. (Microsporon) Audouini Gruby, Compt. rend. Acad. Paris 17:301-303. 1843. 1916] Emic—Yerast FUNGI 269 Sixty to 65 per cent of the fungous affections of the scalp are caused by Sporotrichum of Microsporon, including eleven varieties which are divided by Sabouraud into two groups: those of the Sporotrichum Audowm type which give slow- growing cultures on artificial media, and those of animal origin which yield rapidly growing cultures. The disease, frequently derived from domestic animals, rarely attacks the glabrous skin, and contagion is more frequent from case to ease. At least four varieties of Sporotrichwm are parasites common to man and animals. Upon microscopic examination of the diseased hair, the fungus appears in crowded cells, 2-3» in diameter, irregu- larly arranged so as to form a continuous covering of the hair without penetrating the cuticle. In the interior we find delicate parallel filaments of large cells. In infection with Microsporon the growth of the fungus progresses from the tip of the hair to the lower parts. S. Furfur (Robin) басс. Syll. Fung. 4:100, 1886. An affection of the cuticle, called ‘‘tinea versicolor,” is characterized by the yellowish or brownish discoloration of the lesions, which at one time were classed with the group of pigmentary stains. The color of the lesion is subject to great variation, not merely in different patients, but in dif- ferent regions of the same patient. This affection caused by S. Furfur was first discovered by Hichstedt in 1846. Little is known of the mycological characters of this parasite. S. (Microsporon) minutissimum басс. in Gedoelst, Les Champ. Par. 1902. An epidermomycosis, erythrasma, presenting some points of resemblance to tinea versicolor, is characterized by brown- ish scaly patches which appear usually in the genitocrural region. The elements of this species are very small, and in preparations, appear as spores and threads of mycelia ar- ranged almost in the same manner as the elements of 8. Furfur. The recognition of sporotrichial infection other than those occurring in skin diseases, is of recent date. The infection, [VoL. 3 270 ANNALS OF THE Missourt BOTANICAL GARDEN in a case described by Schenk in 1898, started in the index finger and led to the formation of a series of subcutaneous abscesses connected by a chain of chronic lymphangitis along the arm. This new parasitic fungus was obtained by cul- tures from the lesions. In 1906 Beurmann (cited by Pinoy, '11) again called attention to this parasite occurring in mul- tiple, widely distributed, gummatous lesions. Sporotrichwm affects not only the skin and subcutaneous tissue, but also the mucous membrane. Intramuscular and periosteal gum- mas, and even pulmonary abscesses may be caused by this fungus. GLENOSPORA G. Graphii ТИЕТИН Vuillemin, Compt. rend. Acad. Paris 154:141-14 Verticillium cie Hass & Bezold, in Siebenmann, Die Schimmelmykosen d. Ohres 95. 1889. This fungus has been reported by Hassenstein, Steudener, Bezold, and Siebenmann as occurring in otomycosis. In seven eases of otomycosis Verticillium has been incriminated four times. This botanical classification of the fungus was not certain, and has not been verified in later investigations. Siebenmann considers Steudener's T'richothecitum, Hallier's Stemphylium, and Harz and Bezold's Verticillium, as identi- eal organisms. Vuillemin (212) followed the development of the above fungus in eulture and was able to explain the diver- gent opinions as to its identity. No degree of regularity in form and position is attained by the conidia, which are dark, one-celled, and irregularly inserted on the mycelium like the eonidia of Glenospora Berk. & Curt. "This then eliminates as parasites of man the genera Stemphylium, Cephalothe- cium, Verticillium, and Ше pseudo-genus Graphium. The species of fungi mentioned above may be summarized in the following manner: PHYCOMYCETES MUCOR M. cornealis Saec. is M. corymbifer Cohn. M. corymbifer Cohn has proven toxie for rabbits and 1916] Еміс--Ткавт F'UNGI 271 guinea-pigs. It occurs frequently as a saprophyte. The occurrence of M. Mucedo L. in instances of mycosis is doubtful; it is non-pathogenic for animals. M. Regnieri Lucet & Cost. is M. corymbifer Cohn. M. Truchisi Lucet & Cost. is M. corymbifer Cohn. M. parasiticus Lucet & Cost. (басс. & Syd.) is Rhizopus nigricans. M. pusillus Lindt has not been reported in cases of mycosis. The occurrence of M. racemosus Fres. in cases of mycosis is doubtful. It is non-pathogenie for animals, and those in- stances of mycosis in which M. racemosus Fres. were re- ported are probably due to Aspergillus fumigatus Fres. M. ramosus Lindt is M. corymbifer Cohn. M. septatus Bezold, as reported in cases of mycosis, is M. racemosus Fres. RHIZOPUS В. niger Ciaglinski & Hewelke is R. nigricans. В. Солт Berl. & De Toni has not been reported in cases of mycosis; it is toxic for animals. MORTIERELLA The determination of Costantin that a species of Morti- erella was present in a mycosis of a cat has not been ac- cepted. ASCOMYCETES ASPERGILLUS A. aviarius Peck is A. fumigatus Fres. A. bronchialis Blum. is A. fumigatus Fres. A. candidus Link has been reported as a saprophyte. A. flavus Link is also a saprophyte. A. fumigatus Fres. is frequently found in lung mycosis, more particularly in birds. Most cases of mycosis are due to this species of Aspergillus. A. fontoynonti Guég. is a parasitic fungus that produces epidermal lesions. It is toxic for animals. Fungi reported under the name of A. glaucus Link are A. fumigatus Fres. [Vor. 3 272 ANNALS OF THE Missourt BOTANICAL GARDEN A. malignus Lindt is A. fumigatus Fres. A. nigricans Cooke is A. fumigatus Fres. А. nigricans Wreden is Sterigmatocystis nigra Van Tieg. А. repens (Corda) Sace. is a saprophyte and the same as A. glaucus Link. A. Tokelau Wehmer is a true parasite. STERIGMATOCYSTIS S. nigra Van Tieg. is a saprophyte and non-pathogenic for animals. SACCHAROMYCES S. anginae Achalme E Troisier and S. granulatus Vuille- min & Legrain are the only two species of Saccharomyces re- ported as animal parasites. SACCHAROMYCOPSIS S. guttulatus (Robin) Schiónning is normally a saprophyte. ENDOMYCES E. albicans Vuilemin produces the disease known as thrush. FUNGI IMPERFECTI Слага ОСОСС Си There are а large number of eryptocoeci that have been deseribed, most of them having been obtained from tumors. Their morphology and botanieal classification have not been sufficiently established to permit a satisfactory summary at the present time. OOSPORA AND SPOROTRICHUM Certain species of Oospora and Sporotrichum are produc- ers of skin diseases. GLENOSPORA G. Graphii (Siebenmann) Vuillemin is evidently a sapro- phyte. EXPERIMENTAL DATA In a preliminary set of isolations a number of yeast cul- tures were obtained from certain fruits, the sap of trees, and the seeds of various plants growing in the Missouri 1916] Еміс--Үкавт FUNGI 273 Botanical Garden and vicinity. The finding of yeast-like organisms on the seeds of native plants at once suggested the possibility of obtaining yeasts from foreign sources. Four hundred and ninety-three samples of seeds of in- digenous plants were received from botanical gardens of Europe, Asia, Africa, Australia, West Indies, Central America, South America, and the Oceanic Islands. A quantity of each kind of seed was separately placed in sterilized test-tubes containing sterilized distilled water. These test- tubes were set aside for twelve hours and then gelatin plates were made by inoculating tubes of sterile beer-wort gelatin with a mm. platinum loop of the water in which the seed had been standing. Only two yeast-like fungi were obtained from these seeds, although a large number of different moulds and bacterial colonies were isolated. The presence of so few yeast-like fungi may have been due to the dry condition which is unfavorable for the survival of many such organisms. Because so few yeasts were obtained from foreign sources, the investigation was later restricted to the material available in and near St. Louis. Cultures of yeasts were obtained by plating from beer- wort gelatin inoculated with minute quantities of infected fruit juices. Pieces of various fruits and the basal portions of flowers were placed in test-tubes of sterilized water for about twelve hours. Pieces of fruits were also placed in moist chambers for about three days or more. In either ease gelatin plates were made by direct inoculation from the moist material. Holes were made in the trunks of a limited number of trees by means of a sterilized auger, one-eighth inch in diameter and extending a short distance beyond the cambium. After three days, samples of the tree sap that had collected in the cavity were transferred to test-tubes of sterilized distilled water by means of a sterilized platinum wire. Gelatin plates were then made directly from the water suspension after it had been standing for about twelve hours. Approximately 850 different sources were examined for yeasts. In all, 180 different strains of fungi were finally obtained from herbaceous seeds, fruits, fruit juices, sap of [Vor. 3 274 ANNALS OF THE Missourt BOTANICAL GARDEN trees, and the nectar of flowers. A second and a third series of plates were made from the original colonies, and in most cases this was sufficient for obtaining pure cultures. Beer-wort gelatin had many advantages over other media, such as beer-wort agar, glucose agar, and potato agar, which are often used for the isolation of these organisms, On agar the growth of bacterial colonies is more rapid than that of the yeast colonies during the first period of incuba- tion; on gelatin the reverse is true, especially with an in- crease of acidity and a higher percentage of gelatin, The beer-wort gelatin used in these experiments was made by diluting hopped beer-wort with an equal quantity of distilled water, and then adding 12-15 per cent gelatin; the final reaction of the medium was +5 to +7. The 180 different yeast fungi finally acquired were tested for growth on blood serum medium at blood temperature. It was impracticable to make inoculation experiments on animals with all of these cultures. The blood serum test, therefore, was made with the purpose of eliminating all the cultures that would not grow at 37°C. and presumably under some of the conditions that are to be met with when an organism is introduced into the blood system of higher animals. The blood serum medium was made by adding one part of nutrient bouillon, obtained from veal, with 1 per cent dex- trose, to 3 parts of ox blood serum. The medium was then sterilized at 60°C. for one hour on five successive days and finally coagulated in an inspissator at 75°C. All the tubes were incubated at 37°C. for twenty-four hours in order to eliminate any that were not sterile. The yeast cultures were then transferred to the blood serum medium and kept at 37°C. in a moist chamber for ninety-six hours. The growth appearing after this time was examined micro- scopically to see if there was any contamination by bac- teria. Of the 180 cultures thus tested only twelve strains were obtained which grew under these conditions. These twelve strains included three red, five white, and four black 1916] Еміс--Үкавт Funai 275 yeast-like fungi. The sources from which they were de- rived were as follows: Culture No. 1. Torula sp? from the sap of Halesia caro- lina. Culture No. 2. Torula sp? from the sap of Pinus sylvestris. Culture No. 3. Torula sp? from the nectar of Salvia splen- dens. Culture No. 4. Torula sp? from elderberry wine spon- taneously fermented. Culture No. 5. Torula sp? from the nectar of Oenothera sp? Culture No. 6. Torula sp? from the seeds of Zea Mays. Culture No. 7. Alternaria sp? from the fruit of Ribes Gros- sularia. Culture No. 8. Alternaria sp? from the sap of Pinus aus- triaca. Culture No. 9. Oospora sp? from the sap of Morus alba. Culture No. 10. Alternaria sp? from the sap of Populus tremuloides. Culture No. 11. Alternaria sp? from the seeds of Sorghum vulgare. Culture No. 12. Alternaria sp? from the seeds of Rhus glabra. Culture No. 13. A variety of Saccharomyces cerevisiae ob- tained from “Yeast Foam." This culture was used as a control in all the physiological experiments. Culture 1, Torula sp?—This culture, obtained from the sap of Halesia carolina, when grown on beer-wort gelatin and beer-wort agar, devel- ops into spherieal colonies of a deep red color, 1-2 mm. in diameter. It may be dis- tinguished from cultures 2 and 3 by its growth on yeast- water agar, on which the col- onies attain the size of 2 mm. Fig. 1. Culture 1. Cellsx 2000. in diameter—that is, five to ten times the size of the other two [Vor. 3 276 ANNALS OF THE Missovumi BOTANICAL GARDEN red cultures of T'orula. It liquefies gelatin rapidly, but culture 2 liquefies gelatin very slowly, and culture 3 not at all. Growth takes place by budding, as in the true yeasts. The cells (fig. 1) are oval or spherical, varying in size from 3-4 X 4-би. In sugar nutrient solutions the organism develops rapidly, with the formation of an abundant sediment of yeast cells; but the film, if any, is very thin and made up of colonies loosely connected. In acid yeast-water solution a yellow- brown sediment is formed except in the presence of malic acid in which ease the red pigment has practically disap- peared, the sediment being almost white. No spores are formed on moist porous plates or on Gorodkowa’s test medium, although in many cells there appeared two to four fat globules that are much like endospores in appearance. Culture 2, Torula sp?—The cells of this strain (fig. 2) vary in size from 3-3.5 X 4-6». Growth is by budding from all sides of the mother eell and without the formation of mycelium. Devel- opment at 37°C. is more rapid than at room tempera- ture. Cultures grown in yeast wa- Fig. 9. Culture 9. Cellsx 2000. ter with the addi- tion of saccharose, glucose, levulose, maltose, and lactose, produce a heavy red deposit of yeast cells and a slight ring formation on the surface of the liquid. In yeast water con- taining 1 per cent of acids—citrie, malic, tartaric, and succinic acids—development is as good as in the sugar nutrient solu- tions. On yeast-water agar the colonies are very small, elliptical or oval in outline, and .2 mm. in diameter. Gelatin is slowly liquefied. No spores are formed. Culture 3, Torula sp?—This organism, obtained from the nectar of Salvia splendens, is brownish red in color. The cells 1916] Еміс--Үкавт FUNGI 277 (figure 3) are oval, 2.84 3-6u. In acid yeast-water solution growth is very rapid; and in the presence of lactic and citric acids, only isolated colonies are forme on the bottom of the culture flask. In sug- ar nutrient solutions Fig. 8. Culture 3. Cellsx2000. it produces a sedi- ment of yeast cells and a thin film on the surface of the liquid. No spores are formed. Culture 4, Torula sp?—This white species of Torula, ob- tained from elderberry wine spontaneously fermented, de- velops in the same manner as the cultivated yeasts. The cells (fig. 4) are oval, 3-4 X 4-5» in diameter. Cultures in yeast water containing saccharose, dextrose, levulose, maltose, and lactose, develop rapidly, with a sediment of yeast cells and the formation of a thin film on the surface of the culture liquid. In yeast water containing succinic, citric, or acetic acids, the film on the surface of the culture liquid is white, opaque, and wrinkled. In the presence of tartaric and Fig. 4. Culture 4. Cellsx 2000. [Vor. 3 278 ANNALS OF THE MissoURI BOTANICAL GARDEN malic acids the separate colonies are connected into a net- like film. Gelatin is not liquefied, and no spores are formed. Culture 5, Torula sp?—The cells of this species (fig. 5), ob- tained from the nectar of Oenothera sp?, are oval to elliptical, 2-2.5 >x 45. On beer-wort agar and gelatin the colonies are small, white, and spherical. In sugar nutrient so- lutions and in yeast water containing citric, malic, or lac- Fig. 5. Culture 5. Cells 2000. tic acids, there is a considerable sediment of yeast cells and a thin film on the surface of the culture liquid, In yeast water containing tartaric acid the colonies remain more or less distinctly formed on the bottom of the culture flask. No spores are produced, and gelatin is not liquefied. Culture 6, Torula sp?—The colonies of this species of Torula, obtained from the seeds of Zea Mays, may be dis- tinguished from cultures 4 and 5 by the growth appearance on agar and gelatin. The margin of the colonies is irregular and extends radially from % the center of growth. The Се cells (fig. 6) are smaller, being 1.5-2.5 X 2-3.5y. Multiplieation is by bud- ding, more particularly at 3 the ends of the mother cells. In nutrient solutions Fig. 6. Culture 6. Cellsx 2000. containing galaetose and levulose, development takes place rapidly, with the ^ "mation of a sediment of yeast cells and a very dense wrink.. ` film on the surface of the liquid. In nutrient solutions containing maltose and lactose, the film is 1916] Emic—Yerast FUNGI 279 made up of a network of distinct colonies, but in the presence of saccharose the film is slimy and stringy in appearance. Dextrose and levulose are fermented. Gelatin is not lique- fied, and spores are not formed. Culture 7, Alternaria sp?—Colonies of this organism, ob- tained from the fruit of Ribes Grossularia, on gelatin or agar plates, appear much like the colonies of wild yeasts. At first they are made up of budding cells; later a few radiating strands of mycelium appear, with budding conidia at or near the cross walls. The culture soon becomes black from the formation of chlamydospores. The growth on Fig. 7. Culture 7. Vegetative cells, conidia, chlamydospores, and muriform spores, agar after seven days, results in the appearance of four distinct regions to each colony. The central part is black and shiny with a slightly wrinkled surface. About this area are three zones concentrically arranged and of a very dark green color, the terminal margin being made up of a filamentous growth. In nutrient sugar solutions, the film is at first white then black. Dextrose and saccharose are fermented. In yeast [Vor. 3 280 ANNALS OF THE Missourt BOTANICAL GARDEN water containing organic acids, there is a sediment of yeast cells and a film of anastomosing colonies. Gelatin is liquefied. The cells (fig. 7) vary in size from 6-8 X 8-10». Besides the single-celled chlamydospores, there are occasionally many- celled spores borne on short conidiophores. The septa in the spore occur both at right angles and parallel to the long axis and are muriform. Culture 8, Alternaria sp?—The colonies of this culture are at first made up of a central mass of yeast cells with my- Fig. 8. Culture 8. Vegetative cells, conidia, chlamydospores, and muriform spores. celium extending radially from the center. Budding conidia are rapidly formed at regular intervals on this mycelium. The cells (fig. 8) vary in size from 3-6 X 4-9, After seven days’ growth on agar, the colonies may be separated into a central flat dise and several distinct zones, these zones gradually changing from a dark green to a black color. In sugar nutrient solutions there is a rapid growth and a very thick, slimy, greenish black film. Dextrose, saccharose, and 1916] Emic—Yerast FUNGI 281 maltose are fermented. In yeast water containing organic acids, growth is as rapid as in the sugar nutrient solutions; but instead of forming a film, the growth on the surface is limited to the margin along the side of the culture flask. The cells vary in size from 2 X 10-25& or 3 X 7-8и. Gelatin is liquefied. Chlamydospores and muriform many- celled spores oceur in acid yeast-water solution. Culture 9, Oospora sp?—The colonies of this organism ap- pear much like certain white yeasts. They are circular in outline and have a more or less wrinkled surface. Growth of this organism does not take place by budding; but the mycelium, which is dichotomously branched, predominates V CM Fig. 9. Culture 9. Vegetative cells. in all media. The cells (fig. 9) are elongated, 2-2.5 X 12-15и, or short and rectangular, 3-446, In yeast water containing organie acids, certain of the cells may become quite large and siekle-shaped, 4—5»x(20-30& Мо chlamydospores or conidia are found in any of the cultures. When grown in Raulin's nutrient sugar solutions the liquid becomes bright red in color. This characteristic easily distinguishes this organism from the other cultures with mycelial growth. Gelatin is liquefied. Culture 10, Alternaria, sp?—This organism growing on agar forms a membrane of yeast cells and mycelium of a flesh-pink color. The yeast cells (fig. 10) vary in size from [Vor. 3 289 ANNALS OF THE Missourt BOTANICAL GARDEN 4-8 X 5-8. The culture does not become dark in any media and rarely forms chlamydospores. In yeast water containing organic acids only a few chlamydospores are found among the cells on the surface of the liquid. In nutrient sugar solutions and in yeast water containing organic acids the Fig. 10. Culture 10. Vegetative cells, conidia, chlamydospores, and muriform spores. vegetative growth results in the formation of a white, glisten- ing mass of fungous elements that cannot be separated easily. Only a few muriform many-celled spores are found, which indicates its being a species of Alternaria that does not readily form spores in culture. Culture 11, Alternaria sp?—This organism liquefies gelatin more rapidly and becomes black—due to the formation of chlamydospores—more quickly than any of the fungi pre- viously described. On gelatin or agar it first appears in irregular yeast-like colonies. The budding conidia develop rapidly; and if the colonies on the surface of the media 19161 Emic—Yerast Funct 283 are crowded, only yeast cells are formed. Saccharose and maltose are fermented. In yeast water containing citric, tartaric, or succinic acids, the sediment is made up of yeast cells, whereas the black, leathery film is composed of mycelia and chlamydospores. The cells (fig. 11) vary in size from 4-6 X 10-12» or 2 x 10-504. The many-celled muriform spores are found in cultures grown in acid yeast-water solu- tions. Culture 12, Alternaria sp?—This fungus is differentiated in part from the other black yeast-like fungi by the late ap- Fig. 11. Culture 11. Vegetative cells, chlamydospores, and muriform spores. pearance of chlamydospores and the predominance of yeast cells. In early cultures on agar the colonies are yeast-like in appearance, and the mycelium starts to develop only after the medium has become less moist. In nutrient solu- tions containing sugars the growth is very rapid, and a ring of colonies appears on the surface. In yeast water containing organic acids there is a sediment of yeast cells, or the growth is limited to isolated colonies on the bottom of the flask. The ring appearing on the surface of the liquid is white in color except in the presence of lactic acid, in which case it is black. This fungus, a species of Alternaria, has dark chlamydospores and many-celled muriform spores. The cells (fig. 12) vary in size from 3-4 X 9-60и. Gelatin is [Vor. 3 284 ANNALS OF THE Missourt BOTANICAL GARDEN rapidly liquefied. Saccharose, dextrose, levulose, and mal- tose are fermented. Culture 13, Saccharomyces cerevisiae—This yeast, ob- tained from “Yeast Foam,’’ has spherical cells (fig. 13) vary- ing in size from 5 to Әріп diameter. Growth takes place by bud- ding from all sides of the mother cell. In nutrient sugar solu- tions the organism forms a sediment of yeast cells and a thin film only in the presence of glucose. In yeast water containing organic acids, the sediment of yeast cells has a floceulent appearance. Saccharose, dextrose, levulose, and Fig. 12. Culture 12. Vegetative cells, conidia, chlamydospores, and muri- form many-celled spores. maltose are fermented, with the formation of aleoliol and carbon dioxide. Spores are formed on moist porous plates, on Gorodkowa’s test medium, and on yeast-water agar. On the latter medium, after a growth of ten days, 30-40 per cent of the yeast cells have formed endospores. The number of ascospores in the asci varies from 1 to 4, and they vary in size from 2.5 to 4и in diameter. Upon germination the spores enlarge and are set free from the spore case and then develop separately, or the spores may fuse as they become larger, the spore case becoming thinner at the same time, which results in a very large cell that develops by budding. Sometimes the ascospores start to germinate be- fore they are set free from the ascus. The reaction of the thirteen organisms was tested in nutrient solutions of saecharose, dextrose, levulose, and mal- 1916] Ewic—Yzasr F'UNGI 285 tose. The sugar nutrient solutions contained 1 per cent peptone, .3 per cent monopotassium phosphate, and .02 per cent magnesium sulphate. The concentration of the sugar was 5 per cent saccharose, D per cent dextrose, 1 per cent levulose, and 1 per cent maltose, respectively. For the determination of alcohol formation, 100 ce. of the sugar nutrient solutions were placed іп 125-ce, Erlenmeyer ^ COQ Fig. 13. Culture 13. Vegetative cells, asei with ascospores, and spores dur- ing germination. flasks, and after sterilization each was inoculated with a pure culture of one of the thirteen different strains and incubated at room temperature for thirty days. Immediately after the inoculation of the nutrient solutions the cotton plugs in the culture flasks were replaced by one-holed rubber stoppers, through which passed a piece of small glass tubing with a small cotton plug. This precaution was taken to prevent excessive evaporation. After the incubation at room temperature for thirty days, the nutrient solutions were neutralized with normal caustic soda and made up to the [Vor. 3 286 ANNALS OF THE Missourt BOTANICAL GARDEN original volume with distilled water. The loss by evapora- tion did not exceed .3 ce. in any case. Fifty се. were then distilled off, and the amount of alcohol present determined with a pyenometer. The presence of aleohol was confirmed in each case by means of the iodoform test as used by Will (710) in his investigation on certain species of Mycoderma. TABLE I SHOWING THE PERCENTAGE, BY VOLUME, OF ALCOHOL PRODUCED BY THE THIRTEEN DIFFERENT CULTURES AFTER INCUBATION AT OOM TEMPERATURE FOR THIRTY DAYS Iodo- Iodo- Iodo- Iodo- үе SE form ман form | — | form | Maltose | form | test test test test 1 .10 —* .10 — „10 -- 30 -— 2 05 — ‚05 — .10 — 20 -— 8 .00 — .10 -- 10 -- 00 -- 4 15 -- .00 - 10 — .20 -- 5 00 -- 00 ЕЕ 10 05 -— 6 .00 -— .85 + 80 + .10 -— 7 .95 4 .65 -- 15 -- 25 - 8 1.20 -р 45 + .10 — 45 + 9 00 — 00 - 05 -- .20 — 10 .10 — .10 -- .05 -— .00 -— 11 .95 + ‚85 -- 115 -- ‚55 + 12 80 + 1.00 + .50 + „75 + 13 5.35 + 5.00 + „70 + .85 + Control .00 -- 00 -- 00 -- 00 — (without TE ism) *The sign + indicates that the iodoform test for aleohol was positive, with the presence of iodoform erystals. The results, as indicated by table r, show that very little aleohol, if any, is formed by the various eultures with the exception of culture 13 (Saccharomyces cerevisiae), whieh was used as a control Only one culture of Torula, culture 6, produced alcohol by the fermentation of dextrose and levulose, Four of the black yeast-like fungi-cultures, 7, 8, 11, and 12, produeed only small quantities of alcohol. The reaction of the thirteen different cultures was tested in yeast water containing organic acids. Seventy-five grams of press-yeast were ground up with enough distilled water to form a thin paste. More distilled water was then added 1916] Emic—Yerast FUNGI 287 to make up the volume to one liter. This yeast suspension was then heated in an Arnold sterilizer at 100°C. for one hour. After most of the yeast cells had settled to the bottom of the flask, the supernatant liquid was decanted and filtered through hard filter paper. This filtrate was then run through a Berkfeld cylinder to remove all yeast cells. The acid nutrient solutions were made from this last filtrate by the addition of 1 per cent citric, 1 per cent lactic, 1 per cent malic, 1 per cent succinic, 1 per cent tartaric, and .5 per cent acetic acid. For the acid reaction 50 се. of the acid nutrient solutions in 100-ee. Erlenmeyer flasks were sterilized, and then each was inoculated with a pure culture from one of the thirteen fungi. After incubation for thirty days at room temperature, the solutions were made up to the original volume with dis- tilled water. A 10-сс. portion was then titrated with N/10 caustic soda, phenylphthalein and litmus being separately used as indicators. The results given in table п are in terms of cubice centimeters of N/10 caustic soda required to neu- tralize 10 ce. of the culture solutions. [VoL. 5 288 ANNALS OF THE Missourr BOTANICAL GARDEN THAT Te TAKEN PLACE TABLE II SHOWING THE FINAL ACID REACTION AND THE CHANGE IN REACTION AFTER THE CULTURES WERE INCU TED AT ROOM TEMPERATURE FOR THIRTY D Phenylphtha- Culture no. Acid ein Difference та Difference as indicator as indicator Acetic 6.3* -- 5.6 -- Citrie 6.55 -- 5.7 - я | Lactic 4.5 — 3.8 -- Control Malic 6.15 НЕ 5.45 2 Suceinie 6.5 — 5.9 — Tartaric 6.0 — 5.5 — Acetic 6.5 + 2 5.7 + 1 Citric 5.5 --6.0 —3 --6.0 1 Laetie 3.0 —1.5 2.8 —1.5 Mali 3.5 —2.65 2.8 —2.65 Succinie 2.9 —3.6 2.3 —3.6 Tartarie 51 — 9 4.5 —1.0 Acetic 6.7 + A 6.1 + .5 Сите В —6.25 —.3 —6.0 9 Laetie .25 —4.25 --.5 8 айе 8 —5.85 —3 —5.75 Suecinie A —6.2 „б --6.4 'Tartarie 3.8 --9,9 8.3 —2.2 Acetic 6.3 .0 5.7 + .1 Citrie 4.1 --9,45 8.8 —2.4 3 Lactic 3.4 —1.1 2.7 —1.1 alie T —5.45 2 —5.25 Succinic 5 D —.4 —6.3 Tartarie 5.2 — .8 4.5 —1.0 Acetic A --6.2 --.5 —6.1 Citric 45 —6.1 —.3 --6.0 4 Laetie 4 —4.1 —.3 —4.1 alie 85 —5.8 —.3 —5.75 Succinic 8 —6.2 —.3 —6.2 Tartaric 6.2 + .2 5.5 .0 Acetic 6.65 + .85 5.9 + 3 Сите 9 —5.65 2 —5.5 5 Malic 2.65 —1.85 2.0 —1.8 Lactic 3 5.85 —3 —5.75 Sueeinic 4 Е —.3 —6.2 Tartaric 5.7 6 5.0 — 6 «Тһе results are given in terms of ce. f and the e iralize 10 = of the culture solutions; an increase r is indicated in terms of ce. N/10 caustic soda per 10 ce. of the ioni solution N/10 eaustie soda required to neu- hange in reaction, whether 1916] Emic—Yerast FUNGI 289 TABLE II. (Continued) i Phenylphtha- Litman Culture no. Acid - Нети Innen as indicator Difference Acetic 6.65 + 35 5.8 + .2 Citric 8 —6.25 — 4 —6.1 6 Lactic sf --3,85 ‚8 —8.6 Malic 2 5.95 — 4 —5.85 Succinic E! mal — 2 —6.1 Tartaric 5.4 — .6 4.9 — .6 cetie 6.9 + .6 ® + .5 Citrie E! --6.15 — 3 —6.0 7 Lactic 1.6 —2.9 1.0 —2.8 alie I —6.05 5 —5.95 Succinic .0 —6.5 — .6 --6.5 Tartarie 5 --5.5 — 3 --5.8 Acetic "f +1.4 6.8 +1.2 Citric 8 —6.25 — 4 —6.1 8 Lactic 1.6 —2.9 LS —2.6 Mali 2 —5.95 — 5 —5.95 Succinic D —6.5 — .6 --6.5 Тагфатте 59 —5.7 — 3 —5.8 Acetic 6.9 + .6 6.0 + A Citrie 6.8 + .25 6.1 + A 9 Lactic 4.6 + .1 4.1 + .3 Malie 6.0 + .15 5.3 + .15 Succinie 6.3 — 2 5.4 — 5 Tartaric 7.5 41.5 7.0 + 5 Acetic 7.6 +1.3 7.0 +1.4 Citric 5.3 —1.25 4.5 —1.2 10 Lactic За -1.4 2.4 --1.4 Malic 8.3 --9,85 9,5 —2.95 Suceinie .4 .1 — .3 —6.2 Tartaric 6.0 0.0 5.6 + .1 Acetic 6.9 + .6 6.2 4.6 Citrie 2 —6.35 — 6 —6.3 11 Lactic a —3.2 St —3.1 Mali а —5.95 — 4 --5.85 Succinie 0 —6.5 — 6 —6.5 Tartarie E! --5.6 — 2 --5.7 Acetie 7.6 +1.3 6.8 +1.2 || Citric 3.8 —2.75 3.0 --2.7 12 Lactic 3.1 —1.4 2.5 —1.3 Malie 5 —5.65 — 1 —5.55 Suecinie д 8 — A —6.3 Tartarie 5.3 |! — .7 4.8 — .7 cetic 6.6 + .3 5.8 + 2 Citric 5.5 +1.05 4.7 +1.0 13 Lactic 2.3 --9,9 1.8 —2.0 Malic 4.4 --1.75 3.7 —1.75 Succinic 4.2 —2.3 3.8 —2.1 Татбагіс 6.4 + 4 5.8 + 3 [Vor. 3 290 ANNALS OF THE Missourt BOTANICAL GARDEN TABLE III SHOWING THE CHANGE IN ACID REACTION E ABOUT BY THE THIR- TEEN gie = DURING THE INCUBATION AT ROOM RATURE FOR THIRTY DAYS (Results are the averages of the two titrations given in table IT) Culture no. Acetic || Citrie | Lactic Malie Sueeinie | Tartarie 1 + .15* --6.0 --1.5 —2.65 —3.6 — .95 2 + .45 --6.1 --4.3 —5.8 —6.3 —2.2 3 A —2.4 —1.1 —5.35 —6.15 — 9 4 —6.15 —6.05 1 —5. —6.2 + 1 5 + A --5.6 --1.8 —5.8 --6.15 — 4 6 + .25 —6.2 —3.7 —5.9 —6.1 — 6 T + .55 —6.1 —2.85 D —6.5 —5.65 8 +1.3 --6,15 —2.15 —5.95 —6.5 --5.65 9 + D + .3 + .2 + .15 — 35 1.5 10 +1.85 —1.2 --1.4 --9. --6.15 0.0 al + 6 —6.3 --9.15 --5.9 --6.5 --5.65 12 +1.25 —2.7 --1.35 --5.6 —6.3 — .7 13 + .25 +1.0 | --2.1 -1.75 —2.2 +3.5 *In terms of N/10 caustic soda per 10 ce. culture solution. The results, as indicated by tables rr and пт, show that all the eultures produce a change in the reaction of the acid yeast-water solutions after ineubation at room temperature for thirty days. There is a decrease in the acidity of all the acid nutrient solutions except in the presence of acetic acid, in which ease a marked decrease in acidity was brought about only by culture 4. In the case of culture 9, only a slight change of acidity was found in any of the acid nutrient solutions. ANIMAL EXPERIMENTS The experiments on animals were made in the laboratory of the pathological department of the Washington University Medical School, under the direction of Dr. E. L. Opie and Dr. W. S. Thomas, Rabbits and guinea-pigs were inoculated with suspensions of the different yeast-like fungi, with the exception of cultures 3 and 13, either from cultures grown in Hansen’s solution for 12 hours, or from 48-hour cul- tures grown on agar and suspended in Ringer’s physiologi- cal salt solution. In each case 2 ce. were injected into the 'Ringer's solution contains the followi ing: Sodium chloride...... .7 per cent Calcium chloride...... .028 per cent Potassium diei. ... 035 per cent Sodium carbonate. .... .003 per cent 1916] Emic—Yerast FUNGI 291 marginal ear vein of rabbits, or 1 ce. intraperitoneally or subcutaneously in the guinea-pigs. All injections were made under aseptic conditions with sterilized instruments. The animals that died were carefully examined for lesions or abnormalities, the autopsy being carried out with aseptic precautions with sterilized instruments. At the same time a sample of blood was taken from the right auricle of the heart, with a sterilized platinum loop, and transferred to a tube containing sterilized beer-wort. All the principal organs were removed to sterilized Petri dishes, and then small pieces of the liver, lungs, spleen, kidneys, and intestines were placed in tubes of sterilized beer-wort. These tubes were then incubated at 28°C. for seventy-two hours, but were kept under observation for over ten days, and then gelatin plates made from them. Where the organisms had been inoculated into beer-wort and the same procedure used as for the animal tissues, pure cultures were again easily obtained on gelatin plates with all the eleven strains used. At the same time pieces of the different organs were fixed in Bouin’s or Gilson’s fluid, imbedded in paraffin, sectioned, and stained with saffranin and methyl blue, or with Delafield’s hematoxylin and eosin, or according to the method used by Pianeze (796) for the staining of carcinoma tissue. [Vor. 3 292 ANNALS OF THE Missourt BOTANICAL GARDEN TABLE IV RESULTS OF ANIMAL EXPERIMENTS = ech | na ЖЕТЕК ИГЕ EENES: e | te | of ВВ E| eflnëälaälez 5S of E 92 SE SE ЕЕ ЕЛЕ ЕНЕ ЕЕН Е 59 2 өс 55 LETE Ме Ог момЕ sn 47 x SE GE m A ке ы A 1| 1|G-p*|M*| 1 |R*] р* " Killed |........... None Negative | 2 r* M 2| H*| V* Г 56 Unknown | White spots Intestines 1 ung air spaces on liver condensed 2 3| G-p|M| 2] H |S* 35 Unknown None None Negative 4| G-plM| 2| H | P | 49 Unknown None None Negative 5| Ср М| 2| R|P ,....| Kil None None Negative 6| G-p|F*| 4| H | S ....| Killed None None Negative 7| Ср|М| 4 | Н | Р 7 Unknown None Intestines Lung air spaces | condensed š 8| G-p|M| 4|H | Р |17 Unknown None None Negative 9 | С-р | Е 4| Е |Р |...) Killed None None Negative 10 | С-р | F 4“ В | P Killed None None Negative 11 г |M| 5|H | V | 14 Unknown None None Negative 12 г IMI S|R|V |.... Killed None None Negative 4 | 13 r MI SIR IV Killed None None Negative 14| G-p|M| 5ІН|65|... Kill None None Negative 15 | С-р | F 5 | Н | P | 60 Unknown None None Negative 16 | G-p|M| 6ІН |65... Unknown None None Negative 5 | 17 | Ср | М| 6| H | P | 16 Unknown None None Negative 18 Е ЦЕ 6| H | V |... Killed None None Negative 6 | 19 | Gp|M| 7 | Е | Р | 20 Unknown None None Negative BIS MI Tp ТЕС жаран електен DN na tee eaten 7121 | Gp|M| 8| R|P 20 Unknown | None None Negative 22 г |Mj| 8 |b-w] V | DE, eee ҚЫҚ ENEE, ГЕ ПЕНЕН ЕЕ ap 23 г | мо V | 60 Unknown White spots, None Negative on liver 8 | 24 | Gp|F | 9|H | P |... Unknown one None Negative 25 | Ср М| 9 | К |Р Е .|Unknown None None Negative 9 | 26 | G-p|M| 10 | R | P 10 Unknown None None Negative WIL MI ДЕ ee V lal es ee aes EE ee e 27 -p|M|11| К | P | 20 Unknown None None Negative 29 r FI2] НУ... Killed None None Negative 10 | 30 | G-p]M| 12| H| S ....| Killed None None Negative 31 | Ср M | 12| H P | 20 Unknown None Intestines Lung air spaces an lungs condensed 32| Ср М|12 | R|P |. Killed None None Negative 11 | 33 | С-р |M | 12 |b-w| P 3 Unknown None None Negative Su ЯҒ! 3а UM ocu ees Seb ЕКОО il vx E Bee S. E "E guinea-pig; r, rabbit; M, male; Е, female; H, de deme in Hansen solutio uspension in Ringer’s solution; P, intraperitoneal; 8, subeutaneous; V ge D gi Ж. sterilized іп beer-wort. то M. mg РА te - 1916] Еміс--Үкавт Funar | 298 TABLE V SHOWING THE WEIGHT IN GRAMS OF THE ANIMALS WEEK BY WEEK* 5 я — š Remarks Е < 1. ..| 970 gms. after 5 months. 2 Died 9 days after 8th wk 3 ied after 4 wks. 5 day 1 Died after 6 wks. 5 days 5 9 gms. after 4 months 6 gms. after 4 months 7 Died after 7 days. 8 Died after 2 wks. 3 days 9 0 gms. after 4 months 10 520 gms. after 4 months 11 Died after 9 day 12 Killed after 2 months 13 Killed after 2 months 14 0 gms. after 3 months 15 Died after 2 months. 16 0 s. after 4 months дро 0 225| 220| ....| ....| Died after s. 5 days. 18 1670| 1575 1690} 1620| 1750| 1700| 1540| 1680| 1820| 1690 gms. after 4 months. IN 9599 $105... ] 408 ааа o. acu cl ep DM RENS 5 days. c baut ЕУ ka МЫ de ndn ER MN de ing. 23128901. 475... 8968 ec P. ES, л... iui Died after 2 wks D days. 23| 1350| 1400| 1340| 1390| 1430| 1430| 1430] 1460| 1420 1965. gms. after 9 wks. 24|....| 382) 380 325| 375| 375| 390] 410| 415| Killed after 2 months. 25) 500 440) ....| 420| 460| ....| 435] ....] 420| Killed after 2 months. ы ЕЛЕС БУК АЗА d vel isl Did aftee 15 0 days. Е Р 300 ҒАН ЫН Ж Бон Died after 20 days. 28| 87 edm rn sooo] sesch sees] sel Living. 29| 1300 1280| 1240 1295) 1250 1250| 1385| 1350| 1415| Killed after 2 months. 80| ....| 345| 325| 310| 338| 342| 359| 325| 345 Killed after 2 months. ӘҢ... 311 300] 280) | ааа ee ln Died after 20 days. 32 470| ....| 385| 420| 415| ....| 455| 450| ....| 535 gms. after 4 months. EEUU. EE usos БР DE s Died after 3 days. 34] 920| ....| . NE e? ҚМ ER es "ing. "Fach animal was weighed once a week from the time of inoculation to the time of death The results of experiment 1 were negative. In experiment 2 animal No. 2, white spots were found on the liver. These lesions were not caused by the organism injected, for inoculations were taken directly from these areas but no fungous culture was obtained. Rabbits are frequently infected by coccidia, and it is probable that they were the cause of the lesions in this animal. Stained [Vor. 3 294 ANNALS OF THE Missovrt BOTANICAL GARDEN preparations revealed a condensation of the air spaces of the lungs. There were no organisms present and this con- dition was probably due to post-mortem changes such as oedema. In experiment 3 animal No. 7 died after seven days, and the autopsy revealed the intestines very much inflamed. The organism was isolated from the intestines, and stained preparations showed abnormalities in the condensation of air spaces in the lungs. А repetition of this experiment gave negative results. In experiment 4 animal No. 11 died in less than two days from causes that could not be determined. The death of this animal may have been due to the fact that phagocytosis was not sufficiently active, since a repetition of the same ex- periment gave negative results. In experiment 5 the results were negative. In experiment 6 animal No. 19 died after twenty days without evidence of lesions. To test out the production of toxin by culture No. 7, 1 ce. of a four-day culture in beer- wort after sterilization was injected intravenously into a 660-gram rabbit, but the results were negative. In experiment 7 animal No. 21 died after twenty days from unknown eauses. The condensation of air spaces found in the lungs was probably due to post-mortem changes. Animal No. 22, а 780-gram rabbit, was subjected to a 1-ce. sterilized injeetion of a four-day eulture of organism No. 8, with negative results. In experiment 8 animal No. 23 died after two months with- out evidence of lesions other than white spots on the liver which were presumably due to coccidia, since no fungous cul- ture was obtained from this lesion. In experiment 9 animal No. 26 died after ten days from unknown causes. Intravenous injection in an 870-gram rabbit of a l-ee. four-day sterilized culture grown in beer- wort, gave negative results. In experiment 10 animal No. 27 died after twenty days without evidence of lesions or the presence of fungi in the various organs. 1916] Emic—Yerast Funct 295 In experiment 10 animal No. 31 died after twenty days. The organism injected was again isolated from the intes- tines and the lungs. The condensation of air spaces in the lungs of this animal was probably due to post-mortem changes. А repetition of this experiment gave negative re- sults, as shown by animal No. 32 which lived four months without evidence of any injurious effects due to an organism. Animal No. 33 received a Jee, injection of a two-day sterilized eulture of organism No. 12, grown in beer-wort. This animal died in three days. However, a 2-сс. intravenous injeetion of a two-day sterilized culture of organism No. 12, grown in beer-wort, gave negative results. All the remaining twenty-four animals inoculated gave negative results, in that no evidence was found of an in- jurious effect due to the cultures injected. Discussion There has been a diversity of opinion in regard to the im- portance of fungi as agents in the production of infectious diseases. More recent investigations indicate that fungi are of secondary importance in the formation of lesions in animal bodies, and usually appear secondarily in infected tissues. On the other hand, certain species of fungi are to be considered of primary importance in those cases in which they prove toxic to animals if consumed in large quantities on infected foods. It is easy to recognize that the parasitic fungus cannot prosper with the same degree of success on all animal species. Mucor and Aspergillus pneumonomycoses, observed frequently in birds, is rarely found in other animals. Certain varieties of Sporotrichum (Microsporon) which occur on infants, seem to grow with difficulty on animals. Certain varieties of Oospora (Trichophyton) are common to man and other animals, whereas still other varieties of Oospora appear only on man. The changes brought about by the disease-producing organisms in the body are quite varied, differing quite as much as the morphological and cultural characters of the organism when grown outside the body. [Vor. 8 296 ANNALS OF THE Missourt BOTANICAL GARDEN In close connection with the anatomical changes produced in the body a study should be made of the physiological relations of host and parasite, more particularly the con- ditions which predispose the body to attack. Sticker (’00) observed that mycosis in man may be of sporadic or of endemic origin. In the former case weakened individuals suffering from other diseases are attacked. Out of thirty- nine cases of mycosis only five occurred in persons sup- posedly in good health. The endemic disease appears in con- sequence of the patient’s vocation: for example, the pigeon caretaker, hair-dressers in Paris, and the sponge purifiers. The diseases produced by fungi, in proportion to the wide distribution of parasitic species, are of rare occurrence in man. Siebenmann, who investigated the distribution of As- pergillus fumigatus from the literature on otomycosis, dis- covered that, while it appeared in all parts of Europe and America, it was most abundant in India, its frequeney de- pending on the time of the year. The experimental results with injeetions of certain species of the Phycomycetes have been mostly negative. All au- thentie instanees of subeutaneous injections have given neg- ative results, whereas only a few species of the Phycomycetes produee death in animals by intraperitoneal or intravenous injeetions. "The resulting lesions in the rabbits and guinea- pigs vary with the manner of injeetion, the kidneys and mesenteric glands being regularly altered. An injection of a 2-ce. spore-suspension of a non-toxic fungus (Sterigmato- cystis mgra) will result in the animal’s losing 10 to 25 per cent in weight during the first three or four days after the injection; after this period the animal rapidly regains its original weight. If the amount of injection is increased, the animal will die from mechanical causes in about eight days. Sections of the kidneys show that the spores have collected in the glomeruli, where some of the spores ger- minate but no multiplication of cells takes place. The heart and blood remain sterile. Ballin (’08) subjected animals to five-day cultures of cer- tain moulds. He then killed the animals after a few hours 1916] Emic—Yurast Funct 297 and investigated the lungs microscopically. According to this author, if guinea-pigs are allowed to breathe the spores of Aspergillus fumigatus grown on agar plates, they die within seven or eight hours by asphyxia. Upon examina- tion of the lungs of the dead animals, he had no difficulty in finding the spores in hemorrhagic colonies which per- meated the lungs. The same result was obtained with eul- tures of Sterigmatocystis nigra. The germination of the spores takes place in the alveolar septa between the alveoli, and then by means of pressure the germinating spores break through the cell walls and enter the alveoli. Fungous infectious agents are not absolutely deprived of the power to secrete toxie substances, although this toxicity does not seem to be as evident in them as in the bacteria. Lucet was among the first to mention the existence of a thermolabile toxic substance in cultures of Aspergillus fumi- gatus. The earlier investigators assert that the intensity of the toxic action of certain fungi is proportional to the quantity of the fungous spores injected, and in this manner the higher fungi differ from pathogenic bacteria, in which the resultant intensity of toxie action is to a large extent independent of the number of bacteria injeeted into the animal. Сеш (707) found that the toxic action of an aleoholie or ether extraction from cultures of Aspergillus fumigatus was of a specific character. He isolated a culture of Aspergillus from the atmosphere in the home of a family affected with chronic pellagra, and obtained a water-soluble toxin from the cultures of this fungus. Не also isolated two toxic varieties of Penicillium from corn, the toxic substance from one variety producing neuromuscular condensations in ani- mals, whereas the toxic substance in the other produced nervous depression. Bodin and Gautier (’06) also found that this toxin was not destroyed by heating to 120°C. for thirty minutes. Otto (’07) obtained an alcoholic extract from five strains of Aspergillus fumigatus which was toxic for animals either by intraperitoneal injection, or as an emulsion if washed into the stomach by means of a probe. Not only [VoL. 3 298 ANNALS OF THE Missourt BOTANICAL GARDEN the spores but also the mycelium contained substances of intensive poison. Sturli (710) extracted a toxin from cultures of Penicillium glaucum, which was neither a phenol, an acid, . or an alkaloid. Blakeslee and Gartner (713) found that the ‘‘presssaft’’ from the aérial filaments of Rhizopus nigricans caused almost instant death when injected intravenously into rabbits. Several other species of Mucorineae were tested but with neg- ative results. A solution containing the water-soluble sub- stance extracted from .045 grams of the dry fungus, when injected intravenously, is sufficient to kill a 1.35-kilogram rabbit in less than two minutes. The poison from Rhizopus appears to be 5.5 times that of the tubercle bacillus, 15 times that obtained from edestin, and 45 times that of penicillie acid. Rhizopus nigricans is widely distributed, and is almost certain to appear as a spontaneous infection on bread and similar substrata rich in carbohydrates when- ever the proper temperature and moisture requirements are observed. Blakeslee and Gartner point the possible relation of this fungus to diseases of unknown origin, such as pellagra, horse disease, and the cornstalk disease of the Middle West, all of which have been attributed to infected food. Mohler (714), in a review of the investigations on cere- brospinal meningitis (forage-poisoning), emphasized the widely aecepted theory that this disease may be due to fungi on the feed. While most investigators have obtained negative results, Mays reported that a colt fed ex- perimentally upon some of the mouldy corn which was held responsible for the serious outbreak in Kansas in 1890, developed the disease on the twenty-sixth day. Again, the Kansas outbreak in 1906 was said by Haslam to have been caused by the consumption of immature ears of corn in- fected by moulds, although the exact mould was not de- termined. By feeding horses upon this corn, typical fatal cases of staggers were produced in four out of seven cases. This theory that toxic fungi cause forage-poisoning is not antagonistic to the facts in many of the more carefully ob- served outbreaks. The great variation in fungous growth 1916] Emic—Yerast Fuxar 299 under different moisture conditions may explain the irregu- larity of the symptoms, as well as the occurrence of the disease under what may appear to be identical conditions. Many horses died as a result of eating mouldy baled hay, and as soon as this hay was eliminated the deaths ceased. Forage-poisoning, therefore, seems to be an auto-intoxica- tion, due to certain chemical poisons or toxins formed by organismal activity. Ruhl (714), in a summary of the new theories concerning the etiology of pellagra, pointed out that investigators ex- plain the casual relation of a corn diet to pellagra by four different physiological processes. However, these theories are inadequate, and serious objections are given to each. In this résumé we find that pellagra epidemics occur among people who have eaten corn that has been previously steamed, whereas persons in the same vicinity who have not eaten corn so treated were apparently not affected. It may be pointed out here that the steaming of corn before grinding will introduce conditions of moisture that favor the de- velopment of fungi, particularly Rhizopus nigricans and cer- tain species of Aspergillus. In a moist condition the corn will become infected with fungi in a few days time; and after the corn has been dried, this fungous growth would no doubt pass unnoticed by the Italian peasants. In the consideration of eryptocoeci, we do not find indica- tions of toxic substances produced by these organisms. Ga- leotti and Pentimalli (’10) investigated the action of yeast toxin on the tissues of higher animals with three cultures of eryptococci obtained from tumors. The injection of the filtrate from liquid cultures or the injection of dead cultures gave negative results. However, the injection of living cells of certain cryptococci have proved fatal to rabbits, guinea-pigs, and dogs; and the organism seemed to show a selective action for the kidneys, the spleen, and the lungs. Loeb, Moore, and Fleisher (’13) were unable to find an extracellular toxin in cultures of the yeast-like fungus ob- tained from a carcinoma tissue. Death of the animals in this latter case was due to the rapid multiplication of yeast [Vor. 3 300 ANNALS OF THE Missourt BOTANICAL GARDEN cells in certain organs, more particularly the kidneys, which resulted in the blocking of the glomeruli and mechanical injury of the tissues. Cases of eryptomycoses in man have been reported in large numbers and appear to be less rare than they were formerly supposed. Diseases of the skin are observed chiefly among persons living under conditions of uncleanliness, or among those who combine such conditions with a tendency to profuse perspira- tion. The disease does not penetrate into the skin itself, but consists, as Plaut has pointed out, of a simple sapro- phytism of the ineiting agent upon the skin. Certain species of Oospora and Sporotrichum which occur as skin parasites on man are non-pathogenie for other ani- mals. Quincke obtained negative results with the spores of Oospora ( Achorion) by subcutaneous, intraperitoneal, or intravenous injections into mice, rabbits, and dogs. Citron (^05) made intraperitoneal injections of 14-day growths of Oospora in beer-wort suspended in salt solution. Pseudo- tuberculosis of the peritoneum resulted from injections of either the living or the heat-sterilized fungus. The question whether or not there are any species among the many known yeasts which are pathogenic for man and animals has been the subject of observation for some time and has been answered mostly in the negative. All the animal experiments made with true yeasts found in nature have given negative results. Raum (91) and Neumayer (291), working on the pathogenicity of yeasts up to the year 1891, came to the conclusion that cultivated yeasts are non-in- jurious to animals. Raum used ten fungi, including Sac- charomyces cerevisiae, 8. Pastorianus, S. ellipsoideus, and S. turbidans. In only one ease of Neumayer's was an injuri- ous effect on animals manifested. Fischer and Brebeck (294) obtained negative results with ten species of Mycoderma, Monilia candida Hansen, and Torula salmonicolor. Rabin- owitsch (795), with fifty cultures of non-spore-forming yeasts, obtained evidence of an injurious action with seven of these fungi, but only in white rats, and in these cases it was neces- sary to use large quantities. San Felice has been able to 1916] Еміс--Үкавт F'UNGI 301 find but one species of Torula that is pathogenic for animals. Cao (’00) obtained positive results with nine of forty-one cultures of what he called Oidium. These results of Сао appear doubtful, for in many instances where the organisms were regained from the inoculated animals they were present in the brain. It is possible for fungi to live saprophytically in the intestines and after death spread to all parts of the body by a rapid growth in the blood vessels. These results of Cao, therefore, must be questioned as to their validity. Cultures of the twelve yeast-like fungi considered in this paper did not produce death in animals or the formation of lesions. Only three of the fungi were isolated from inoculated animals; and in these cases the organism was found in the intestines, and in only one animal was the ioculated organism found in the lungs. The organism found in the lungs of a guinea-pig may have developed in the blood vessels after death. A repetition with inoculations of these same organisms gave negative results. Conor USIONS From a review of the literature on animal pathology we find that fungi—not including bacteria—are of secondary im- portance in the formation of lesions in animal bodies and usually appear secondarily in infeeted tissues. On the other hand, certain fungi are to be considered of primary importance in those instances in which they prove toxie to animals if consumed in large quantities on infected foods. One hundred and eighty cultures of yeast-like fungi were obtained from 850 different sources, but only twelve of these grow on a blood-serum medium at 37°C. These twelve fungi include six cultures of Torula, five of Alternaria, and one of Oospora. One eulture of Torula and four of Alternaria produced small quantities of alcohol in sugar nutrient solutions. In acid yeast-water solutions all twelve of these organisms bring about a change in the acidity reaction, there being a decrease in the acidity of all the acid nutrient solutions [Vor. 8 302 ANNALS OF THE Missourt BOTANICAL GARDEN except in the presence of acetic acid and in culture No. 4. The results of thirty-four animal experiments were neg- ative, in that the death of certain animals was not caused by the formation of lesions or abnormalities due to the organisms injected. No extracellular toxins were obtained from the cultures of these twelve yeast-like organisms. The results of these experiments and a review of literature on animal pathology indicate either that pathogenic yeast- like fungi do not occur in nature, or that if they are present, they are so few as to be met with only under exceptional conditions, In conclusion, the writer wishes to express his thanks to Professor George T. Moore, for the suggestion of the prob- lem and for numerous courtesies extended during the prog- ress of the investigation while at the Missouri Botanical Garden; to Dr. E. L. Opie and Dr. W. S. Thomas, for sug- gestions and aid in the animal experiments which were carried out at the Washington University Medical School; Professor W. C. Allee, for various suggestions and assistance in certain of the animal experiments; to Dr. Gayfree Ellison, for courtesies extended at the Bacteriological Laboratory of the University of Oklahoma; and to Mr. G. H. Smith of the University of Oklahoma, for assistance in reading the proofs. Graduate Laboratory, Missouri Botanical Garden. BIBLIOGRAPHY Axamit, О. (’07). ee ere en d nach Hefeinjektion. Archiv f. Hyg. 62:15-54. pl. 1. 1907. ат (708). Das Schicksal inhalierter Sehimmelpilzporen, ein Beitrag zur Kennt- s des Infektionsweges durch Inhalation. Zeitsehr. f. Hyg. 60:479—489. 1908. Berthelat, C. J. (203 Les Mueorinées pathogénes et les mucormycoses chez 903. Phomme et chez les animaux. These. 197 pp. Paris ie e R. (796). Ueber da orkommen von Susi uin in den Epithe- omen und ihre parasitüre Bae Zeitschr. f. Hyg. 23:283-305. pl. 4. don A. F., and Gartner, R. A. (713). On the occurrence of a uice preme from the bread mould, Rhizopus nigricans (Mucor ас Bi ol. Bul. 2:542-544. 1913. Blumentritt, F. ('01). Ueber einen neuen, im Menschen gefundenen Asper- c" (Aspergillus bronehialis, n, sp.). Ber. d. deut. bot. Ges. 19:442-446. pl. 2 DEET eme aera sss 1916] Еміс--Үкавт FUNGI 303 — ——- (0 Aspergillus bronchialis Blumentritt, und sein nächster Ver- wandter mis en Fres.). Ibid. 23: 419-427, pl. 19. 1905 Bodin, E., et Gautier, L. ('06 Note sur une toxine produite par l'Aspergillus fumigatus. [ Centralbl. E Pakt. I. 38:700-701. 1906.] TOM M. (701). Sur les propriétés du sérum des cancereux au point de e des anticorps des levures. Ibid. 30:945-948. 1901. Nn O. (703). Die Sprosspilze. In Kolle, W., und Wassermann, A. Hand- bueh der pathogenen Mikroorganismens 1: 661—700. f. 1-8. 1903. SI See i Oidien und Oidiomykose. Zeitschr. f. Hyg. 34:282-340. pl. 8-4. Castellani, A. ('11). Observations on fungi of the зае Endomyees affecting the tropics. Centralbl. f. Bakt. I. 58:236-238. 1 f. 1911. See V. (714). Eine neue Form von Keratomykosis (Keratomykosis mu- corina). Ibid. 72:23-37. pl. 1-2. 1914. Сеш, C. (704). Localizzazione delle spore aspergillari nelle glandole mesen- teriche dei с е Ја loro consecutiva attenuazione. [Ibid. 35:563. 1904.] —— — ——, (7042). Sulle T rar patogene d. Penicilium glaucum nella eti- ologia della pellagra. [Jbid. 1904.] ------, (707). Di una nuova specie di Aspergillus varians e delle sue pro- prietà patogene in rapporto coll’eziologia della pellagra. |1014. 39:562-563. 1907.] ------ und Besta, C. (703). Ueber die Toxine von Aspergillus fumigatus und Aspergillus flaveseens und deren Beziehungen zur Pellagra. [Ibid. 33:408. ] Chantemesse, A. (’91). Eine mykotische Pseudotuberkulose. [Ibid. 10:442. 1891.] Citron, J. (705). Ueber EE Verhalten der Favus- und Trichophytonpilze im Organismus, Zeitschr. f. Hyg. 49:120-134. 1905. Clerc, A., et Sartory, А. (708). Étude biologique d'une levure isolée au cours d'une angine chronique. Compt. rend. Зое. Biol. 64:135-137. 1908. Colpe, үй (794). ei rg als Krankheitserreger im weiblichen Genitaleanal. v f. Gynük. 47:635-645. 2 f. 1894. "ein G., und Friseo, B. (795). Pathogene Blastomyceten beim Menschen. Beiträge zur Aetiologie der bösartigen Geschwülste. Centralbl. f. Bakt. I. 18:368-373. 1895. d J. (792). Note sur un eas de pneumomyeose observé sur un ehat M. Neumann. Bul. Soe. Mye. 8:57-59. 1892. Е ЖА Sur les levüres des animaux. Ibid. 17:145-148. 1901. — F (’0 Recherches sur Vere Aspergillus pathogènes. Ann. d. Sei. Nat., oe IX. 2:119-171. pl. 1905. — Lueet, A. (703). Sur le Sterigmatocystis pseudonigra. Bul. бос. Mye. 19: 23 44. 1903. Curtis, Е. (795). Note sur un nouveau parasite humain, Megalococcus aid а, trouvé dans un néoplasme de la région inguino-erurale. Compt. end. Soe. Biol. 47:715-718. P1895. Fermi, und Aruch, E. (’95). Ueber eine neue pathogene Hefeart und tiber die Natur des sogenannten ее eus fareiminosus Rivoltae. Centralbl. f. вац. І. 17:598600. 4 f. 1 теб B., und Brebeek, C. (794). Zur Morphologie, Biologie und Systematik der Kahmpilze, der Monilia candida Hansen und des Soorerregers. pl. 1-2. Jena, 1894 [Vor. 3 304 ANNALS OF THE Missourt BOTANICAL GARDEN gig cim = P. (776). Beobachtungen ro oe beim Menschen. rehow's Archiv 66:330—365. pl. 15. Galeotti, G., und Pentimalli, F. ('10). Ueber die von pathogenen Hefen und ihren Toxinen erzeugten Neubildungen. Centralbl. f. Bakt. I. 56:312-325. pl. 1-2. 1910. Gedoelst, L. (702). Les champignons parasites de l'homme et des animaux domestiques. 199 pp. аа 1902. Grawitz, P. (277). Beiträge zur systematischen Botanik der pflanzlichen Parasiten mit око Untersuchungen über Ea prs sie bedingten Krankheiten. Virchow’s Archiv 70:546-598. pl. 18 ue ^ iE. CI TOR EE ärztlicher Gessellschaften. Qo klin. Wochen- 7:8-9. 0.] к, (3). Recherches sur la nature, le siége et le développement du Por- rigo deealvans ou Phytoalopéeie. Compt. rend. Acad. Paris. 17:301-303. 1843, ------, (744). —: sur les eryptogames d Е la maladie contagieus se du cuir chevelu décrite sous la nom de me tondante (Mahon) Herpes tonsurans Desen ге). Ibid. 18:583-585. iMi Guéguen, Е. (205). Quelques mots sur les Aspergillus pathogènes. Bul. Soe. ye. 21:243-245. 1905. —— n (08). Les champignons parasites de l'espéee humaine, Rev. Sei. V. 9:745-752. 1908. —— ————, (709). Aspergillus ы nova sp. parasite probable des nodosités juxta-articulaires. Compt. rend. бос. Biol. 66:1052-1053. 1909. Hatch, W. K., and sert SÉ (700). Fungus disease of the ear. The Lancet 2:1561-1564. 1 f. Mire L. (700). Ein neuer и Pilz — Sporothrix von Schenk. Сеп- ralbl. f. Bakt. I. 27:689683. 19 Hiekel, R. (706). Beitrüge zur Morphologie und Phy w des Soorerregers (Dematium EEN rh nt=Oidium Га. Robin). K. Akad. Wiss., Wien, math- rw. Kl., Sitzungsber. 115:159-197. pl. 1-2. 1906. Jakowski, M. jm Otomyeosis mueorina. Centralbl. f. Bakt. I. 5:388-389, 1889. Jensen, V. ('03). Ueber die Entwickelung der dureh subeutane Einimpfung von Saccharomyces neoformans Geer ее hervorgerufenen Knotehen. Zeitschr. f. Hyg. 45:298—308. pl. 4. Köllner, eat Sehimmelpilzerkrankung der Sklera. [Centralbl. f. Bakt. I. 40:432. 07.] Klug, A. (04). Der Hausschwamm, ein pathogener Parasit des menschlichen und tierisehen Organismus, speziell seine Eigenschaft als Erreger von Krebs- gesehwülsten. [Jbid. II. 11: 934— 235. 1904.] Leber, Т. (779). Ker а aspergillina als Ursache von Hypopyonker- atitis. Archiv f. Opthalm. 25':235—301. 1879. ——— ÚS (’82). Ueber die Wachsthumsbedingungen der EEN im menschlichen und thierisehen Körper. Berl. klin. Wochenschr. 19:161-164. 1882. Leopold, G. (’00). Untersuchungen zur Aetiologie des Careinoms und über die pathogenen Blastomyeeten. Archiv f. Супак. 61:77-120. pl. 1-6. 1900. Lichtheim, L. (’82). Ueber pathogene Schimmelpilze. Berl. klin. Wochen- sehr. 19: 129-132, 147-152. 1882. 1916] Emic—Yerast FUNGI 305 = W. (86). Mittheilungen über einige neue ui dd Schimmelpilze. Archiv f. exp. Path. u. Pharm. 21:269-298. pl. 2-3. — ——,(789). eber einen neuen pou zen aus dem men- schlichen Gehórgang. Ibid. 25:257-271. f. 1-11. 1889. Linossier, G., et Roux, G. (799). Sur la МЕРЕКЕ et la icq du cham- pigno n du muguet. Compt. rend. Aead. Paris 109:752-755. 18 Loeb, L., Moore, G. T., und DM M. 8. (713). Ueber das Vorkommen von He fen in pud бн Tum mit Versuchen über das Wachstum einer pathogenen Hefe im TlorkÜrpei. SEET f. Bakt. I. 67:450-472. 1913. Lueet, A. ('01). Contribution à 557 SEET et pathogénique de la langue noire pileuse. Archiv. de Par. 4:262-28 1901. ——nyhh k et Costantin, vi dE 0). UE parasiticus espéce pathogéne de ышк Rev. Gén . 12:81-98. pl. SC Contributions à l'étude des Mucorinées pathogénes. —, -----, Arehiv. d. Par. 4:362-408. f. 1-31. 1901. Maffueei, A., und Sirleo, L. (’98). Ueber die Blastomyeeten RS Infeetion- serreger bei bósartigen Tumoren. Zeitsehr. f. Hyg. 27:1-30. pl. 1. 1898. — L. (710). Sur un nouveau groupe de champignons VHC HER ents des Sporotrichoses. Compt. rend. Acad. Pada 150:543-545. 1910. -~ ——V et Dassonville, C. (798). Sur le champignon de l'herpes на) et les formes voisines, et sur la classification des Ascomycétes. Bul Mee, 15:240-253. f. 1. 1898. —————, -----, (799). Sur la position systématique des DI et des formes voisines dans vi classification des champignons. Compt. rend. Acad. Paris 128:1411-1413. 1899. ----, ——, (799). Sur le Ctenomyces serratus Eidam, comparé aux champignons des Teignes. Bul. Soc. Мус. 15:305-310. 1895. — n — —, (700). Sur une forme de reproduction d'ordre élevé chez les up ne Ibid. 16:201-208. 1900. EL et Ramond (705). Un type nouveau de се pathogéne chez l'homme. SCH rend. Soc. Biol. 59:378-380. 1905 Michailow, S. (711). Zwei neue Fälle von Pilzbefunden im Bereiche des Zen- traln nervensystems. Centralbl. f. Bakt. I. 60:500-511. f. 1-5. 1911. Мол. J. R. (714). Cerebrospinal meningitis (‘‘forage poisoning’’). U. Б. t. Agr., Bul. 65:1-14. 1914. t oM duas (LY. ir he c uei über die Wirkungen der verschiedenen Hefearten, welche bei der Bereitung weingeistiger Getränke vorkommen, auf den thierischen und но еу Organismus. Archiv f. Hyg. 12:1-60. 1891. Otto, M. (707). Ueber die еее GE Stämme von Aspergillus fumi- gatus und Penicillium gla nebst einigen Bemerkungen über Pella- gra. [Centralbl. f. Bakt. I. 39: 1563-565. 1907. Paltauf, A. (785). Mycosis mueorina. Virchow’s Archiv 102:543-564. m! 6. 1885. Peck, C. H. (791). Species of plants not before reported. N. Y. State Mus., Ann. Rept. 44:127-187. 1891. Pianeze, С. (796). Beitrag zur Histologie und Aetiologie des Carcinoms. 1 z. path, Anat. (Zeigler). 1-193. 1896. Pinoy, E. (711). Forme du Sporotrichum Beurmanni dans les lésions hu- maines. Compt. rend. Acad. Paris 152:286-288. 1911. h. —=_ A [Vor. 3 306 ANNALS OF THE Missourt BOTANICAL GARDEN Plaut, H. C. (705). Die Hyphenpilze oder Eumyeeten. Іп Kolle, W., und ene rma A., Handbuch der pathogenen Mikroorganismen 1: 526-660. f. 1-5 1903. Pound, R. (701). An cet to n parasites of the human ear. Am. Mier. бос., Trans. 22:81-88. pl. 18. 190 Rabinowitseh, L. Қыл eg über pathogene Hefearten. Zeitsehr. f. Hyg. 21:11-94. 5. Rajat, H., et Péju, G. E Le parasite = muguet et sa ри dans la clas- sification botanique. Compt. rend. Soc. Biol. 61:617-618. 1907. Raum, J. (291). Zur TOM und ней der Sprosspilze. Zeitschr. f. Hyg. 10:1—50. pl. 1-2. 1891 —— M. (2785). Beiträge zur EEN der Infections-krankheiten. . med. Wochensehr. 11:717-719. 1885. Roger H., Sartor (709). Note sur une E Oospora dius ыы oui. ` Compt. rend, Soe. Biol. 150-152. 1909. : ‚ —— — ——, (7094). Ооврога buccalis. Ibid. 301—303. 1909. Roneali, D. B. (7/95). Die age debi in den Adenocarcinomen des Ovariums. Centr а. f. Bakt. I. 18:353-368. pl. 2. 1895. , (7953). Die amm in den Sarkomen. Ibid. 432—434. 1895. Ruhl, K. (714). Die neueren Lehren über die Atiologie der Pellagra. Ibid. 62:321-340. 1914. San Felice, Е. (795-703). ES e можа тама der Blastomyceten. Г. Zeitschr. f. Hyg. 21:32-58. 394—4 pl. 9-10. 1895-96; Ibid. П. 22:171-200. pl. 2-3. P P vid. ^ 29: 463- 504. 6 4-8. 1898; Ibid. VI. 44:364-396. pl. 6-2. 1903. SR A. (’07). Etude SE pe Cryptococcus (Saccharomyces) glu- s Fres. (Kütz.). Bul. Вос. Myc. 23:87-89. 1907. -------, (’11). Contribution a l'étude de quelques Oospora pathogènes. Ibid. 27:160-171. f. 1-11. 1911. Schubert, P. (785). и, Ger der Aspergillusmykosen. Deut. Archiv f. klin. Med. 36:162-18 1885 Schwartz, С. (706). Ein 2. behandelter Fall von Pneumonomykosis aspergillina. [Centralbl. f. Bakt. I. 38:295. 1906.) Siebenmann, F. (’89). Die зегі i aan des menschlichen Ohres. Wies- baden, 1899. [Cited by Pla *03.] Sticker, G. (200). ШІ der Lunge. Spee. path. u. ther. Not othnagel. 14: Wë 192. 1900. Sturli, А. ('10). Uber ein in Se АР Ce weem glaucum) vorkom- mendes ditt. 'LCentralbl f. Bakt. II. 25:334. 10.] iae i H. = Ueber pathogene GE Ibid. I. 19:105-113. 38-5. 1896 Trier E., et Achalme, P. ('93). Sur une angine parasitaire causée par levure et I amas d ти ап muguet. Archiv. Méd. Exp. d'Anat. Path. 9-37. f. 1 1893. Uhthoff, W. ('83). Beitrüge zur г pathologischen Anatomie des Auges. Archiv £. Opthalm. 29°:167-190. pl. 3-5 Virchow, R. (756 ae ci Moris von den beim Mensehen vorkommenden pflanzlichen с ws Archiv 9:557—593. 1856. Vuillemin, P. (298). Les caractères spécifiques du champignon du muguet (E ndomyees albicans). Compt. rend. Acad. Paris 127:630-633. 1898. 1916] Еміс--Үкавт FUNGI 307 — ————, (719). Sur un champignon parasite de l'homme, Glenospora Graphii (Siebenmann). 1014. 154:141-143. 1912. — — ——, et Legrain, E. Ери Sur un cas de Saccharomyces humaine. Archiv. d. Par. 3:237-268. f. 1-3. 1900. Wehmer, C. (298). Die сенй Aspergillus. enëve, Mém. 33:1-157. 1898. — ———, (704). Der Aspergillus des Tokelau. Centralbl. f. Вакф. I. 35:140- 146. f. 1-9. 1904. kamas A. (778). Beobachtung von Pneumomycosis aspergillina. Wien ochenschr. 28:1290-1991. 1878. Soe. Phys. et d'Hist. Nat., Will, H. : '10). Beiträge zur nee der Gattung Myeoderma. Centralbl. 1910 f. Bakt. II. 28:1-37. 15 f. Annals of the Missouri Botanical Garden Vor. 3 SEPTEMBER, 1916 No. 3 THE MISSOURI AGRIMONIES BENJAMIN FRANKLIN BUSH Having collected some interesting specimens of Agrimonia in 1915, that I could not place satisfactorily by the manuals, I was led to make a closer examination of the species of this genus, and through the kindness of Dr. Moore, of the Mis- souri Botanical Garden, I was enabled to study all the Mis- souri specimens of this genus in the Garden herbarium. For more than sixty years the species of Agrimonia have been but little understood, that is, up to the year 1893, when Britton's *Manual" began to be of influence in the way of specific names. The genus seems to have been very much neglected in Missouri, for I have seen only sixteen specimens collected in the state prior to 1893. This seems rather remarkable in a genus of so many species which are so conspieuous in the field, but the fact is due in large part to the prevailing impression that we had only two Amer- lean species, one of which was also found in Europe, and therefore it was of no importanee to make specimens of such widely distributed species. Such an impression also pre- vailed in many other genera at that time, notable examples of which are Ranunculus repens, Scrophularia nodosa, Amar- anthus Blitum, and Portulaca oleracea. Of the sixteen specimens examined, collected before 1893, six were labeled A. parviflora, five correctly, but one is A. iBritton, N. L. Manual northeastern states and Canada. 1891. ANN. Mo. Bot. GARD., Vor. 3, 1916 (309) [Vor. 8 310 ANNALS OF THE MISSOURI BOTANICAL GARDEN rostellata. Only two specimens of A. rostellata are included in this lot, one labeled A. parviflora, as cited above, and the other being called A. striata. Only one specimen of A. platy- carpa was found and that was labeled A. Eupatoria. Тһе remaining eight specimens are all A. pubescens, of which three are indicated as A. Eupatoria, one A. striata, and four are left blank as to specific name. It will thus be seen that these sixteen earlier collections bear but three names, six being called A. parviflora, four A. Eupatoria, and two being given the name of A. striata, while four collectors did not signify any choice of specific names. It is true that one of these sheets bears a label on which the species is indicated as A. microcarpa, but this specimen came to me unnamed, and sometime in 1893 I af- fixed this name to the specimen, being led to do this by Britton's ‘Manual.’ It is hard to conceive how two such unlike species as A. rostellata and A. pubescens should both be named A. striata by the collectors, but in those days A. striata was little understood, and Michaux’s species was not found in any of the manuals or floras, although it was well characterized and is one of our most distinet species. Several authors have studied and described species and varieties of Agrimonia, notably Solander, Michaux, Muh- lenberg? Pursh, Torrey and Gray, Маго, Britton,’ Bicknell, and Kearney.” Our oldest species is 4. parviflora Solander, a distinet species by itself, and one generally understood. The next species to be described is Michaux’s A. striata, here men- tioned because it was formerly confused with A. rostellata 1Ait. Hort. Kew. 2:129-131. 1789. 2р], Вог. Am. 1:287. 1803. 3Cat. Pl. Am. Sept. 47. 1813. 4Fl. Am. Sept. 1:335—336. 1816. er . 1842. "Bull. Torr. Dot, Club 19:221. 1892. id 896. 9Ibid. 24: :565. 1897. 1916] BUSH—THE MISSOURI AGRIMONIES 311 and A. pubescens, and which may ultimately be found in northeastern Missouri. Muhlenberg, in 1813, indicated A. pumila as a good species but unfortunately did not describe it, and also deseribed A. Eupatoria, var. hirsuta, which eighty-three years later was raised to specific rank. Torrey and Gray, in 1840, described A, Eupatoria, var. mollis, which fifty-two years afterward was described as a species. Wallroth, in 1842, monographed the genus, giving six of our species, one, A. microcarpa, hav- ing been described as A. pumila by Muhlenberg. Most of Wallroth’s names are valid and are the ones that will be accepted in American botany. Britton, in 1892, half a century since any work was done on the genus, described A. mollis, based on the A. Eupatoria, var. mollis of Torrey and Gray. Bicknell, in 1896, raised Muhlenberg’s 4. Eupatoria, var. hirsuta to specific rank, giving at the same time a description of this and a new spe- cies, 4. Drittoniana. He was apparently unaware that Wall- roth had already described the former species as A. gryp- osepala, and that the latter was described by Michaux in 1803 as A. striata. Kearney, in 1897, described a variety, A, mollis, var. Bicknellii, which afterwards was found to be the same as A. mollis, described by Wallroth in 1842 as A, pubescens. Prior to 1893, when Britton’s ‘Manual’ began to influence the names in this genus, but three specific names were in use for species of Agrimonia in the eastern states, A. Eu- patoria, A. parviflora, and A. striata, the latter not being mentioned in any manual or flora of recent date, and the name applied also to both A. rostellata and A. pubescens. Muhlenberg, in his ‘Catalogue,’ recognized and indicated several species and varieties in this genus, but these were not accepted until after 1891, and his names are nomina nuda. Torrey and Gray, in 1840, described several species, among which was A. Hupatoria, var. mollis, afterwards raised to specific rank by Britton in 1892. Wallroth’s mono- graph of this genus, in 1842, was not recognized by any of [Vor. 3 312 ANNALS OF THE MISSOURI BOTANICAL GARDEN the manuals or floras until 1891, when Britton in his ‘ Manual’ accepted some of the species. Wood, in the last edition of his ‘Botanist and Florist,’ in 1876, gives only A. Eupatoria and A. parviflora, while Tracy, in his ‘Flora of Missouri,’ in 1885, catalogues the same species, the latter correct, the former including A. pubescens and A. rostellata. Gray, in the sixth edition of his ‘Manual,’ in 1890, gives but two species, A. Eupatoria and А. parviflora, the latter correct, the former including all the other species. Eggert, in his catalogue,! gives these same two species, A. parviflora being correct, but A. Ewpatoria is applied to A. rostellata. Chapman, in his ‘Flora,’ describes A. Eupatoria, A. Eupa- toria, var, mollis, and A. parviflora, the last two correct, the other being A. gryposepala. Having thus reviewed the history of the species found, or likely to be found, I now present for the Missouri species the following key: L рне. hypanthium with several series of Шы В the lower reflexed; cemes and leaves beneath with loose spreading һаїтз................ EAR Or ZO EE 1. Agrimonia gryposepala 1: Wochen? eege reg with 2—4 series of bristles, the latter erect, ascending, or merely Rn racemes and leaves beneath elosely or softly pubes- airg or gla 2, Racemes and leaves glabrous or nearly so, glandular- -granuliferous ; root ілһегопв-іһісКелей........................ 2. Agrimonia rostellata 2. Racemes and lower surface of the leaves decidedly hairy. 3. m not — leaflets not conspieuously p granu ulifer- в beneath. .............................. Agrimonia parviflora 3. Sep tuberous-thiekened; leaflets not po -granuliferous or only slightly so, velvety- pubescent benea 4. ges hypanthium campanulate or lg rer longer than bro rim of fruit not conspicuous; leaflets of the lower елен 7-13; long hairs of the stem сей от wer DT Ae BEE EE EE RTE 5. Agrimonia pubescens 4. Fruiting erer А we сие, EH as broad as long or broader, shorter the sepals, with a prominent rim; lower leaves with 3-5, be 7, lenflets; long hairs of the stem di- varicate. RENT DE eg 1Catalogue of ri phaenogamus and vascular eryptogamous plants in the vicin- ity of St. Louis ?Flora hinc Жин "United States, 1897. 1916] BUSH—THE MISSOURI AGRIMONIES 313 5. Leaves usually erowded оп Ше lower Zeg? of the stem; pel lets 3 or 5, with the lower pair much reduced, rounde the apex; fruiting hypanthium as broad as long ........ Agrimonia microcarpa G oe o 99 €«€9V49939»4€66€€6»29»94à485»6005U8284645 9 oo 9. Leaves seattered on the stem; rere 5-7, acute at the a BE? fruiting hypanthium as broad a long, or usually bro 4. Agrimonia Poe 1. Agrimonia gryposepala Wallr. Beitr. Bot. 11:49. 1842. Agrimoma Eupatoria Pursh, Fl. Am. Sept. 1:335. 1816, not L. 1753. Agrimonia Eupatoria Gray, Manual, ed. 6, 161. 1890, not ed. 2. 1753. Agrimoma Eupatoria Chapman, Fl. Southern U. S. 133. 1897, not L. 1753. Agrimonia Eupatoria, var. hirsuta Muhl. Cat. 47 (hypo- nym). 1813; Barton, Fl. Phila. Prodr. 53. 1815. Agrimonia hirsuta (Muhl) Bickn. Bull. Torr. Bot. Club 23:509. 1896, not Agrimonia hirsuta Boug. 1842. The only specimens seen are the following: Ethel, Bush 7842, Sept. 23, 1915, good complete plants. 2. Agrimonia rostellata Wallr. Beitr. Bot. 11:42. 1842. Agrimonia Eupatoria Gray, Manual, ed. 6, 161. 1890, not L. 1753. Agrimonia Eupatoria, var. glabra Muhl. Cat. 47 (hypo- nym). 1813. Agrimonia Eupatoria, var, parviflora Hook. Fl. Bor. Am. 1:197. 1840. Agrimonia parviflora Seringe, in DC. Prodr. 2:588. 1825, not Agrimonia parviflora Soland. in Ait. Hort. Kew. 2:130. 1789. Agrimonia americana Lucae, in Wallr. Beitr. Bot. 11: :43, as synonym. 1842. Agrimonia striata Bickn. Bull. Torr. Bot. Club 23:512. 1896, not Agrimonia striata Michx. 1803. The following specimens have been examined: Williamsville, Wayne Co., T'release 149, Sept. 9, 1897, a small, weak, poor specimen, ‘tat undoubtedly this species, in flower and young fruit, Herb. No. 52,255; Swan, Taney Co., [Vor. 3 314 ANNALS OF THE MISSOURI BOTANICAL GARDEN Bush 560, Sept. 24, 1899, a small, weak plant in young flower, Herb. No. 52,256; Monteer, Shannon Co., Bush 746, Aug. 22, 1901, a complete specimen in flower and fruit, Herb. No. 52,257; East Bertig, Dunklin Co., T'release, Oct. 28, 1897, a small, weak plant with fruit all fallen off, Herb. No. 52,254; Meramee Highlands, St. Louis Co., A. G. Johnson, July 29, 1905, the upper part of a robust plant in flower and young fruit, Herb. No. 52,252; Spring River, Jasper Co., Trelease 982, Sept. 18, 1898, a fine, complete plant with nearly all the fruit fallen off, Herb. No, 52,253; Thornton, Clay Co., Mackenzie 642, Oct. 18, 1901, a complete plant with all the fruit fallen off, Herb. No. 52,258; Campbell, Dunklin Co., Bush 110, Aug. 16, 1895, a small, weak plant not yet in flower, Herb. No. 52,259; Dodson, Jackson Co., Bush 107, Aug. 26, 1895, a complete plant in full-grown fruit, Herb. No. 52,260; Flat River, St. Francois Co., T'release, Oct. 13, 1897, a poor, weak plant with the fruit all fallen off, Herb. No. 52,261; Webb City, Jasper Co., Bush 6026, July 23, 1910, a good, complete specimen in flower and young fruit, Herb. No, 52,262; Fern Glen, St. Louis Co., A. G. Johnson, July 14, 1906, the upper part of a plant not yet in flower, with narrower leaves than usual, but evidently belonging to this species, Herb. No. 52,263; Meramec Highlands, St. Louis Co., Eggert, July 10, 1897, a fine, large plant scarcely yet in flower, Herb. No. 52,264; Monteer, Shannon Co., Bush 4896, Oct. 10, 1907, a poorly pressed, large plant in fine fruit, Herb. No. 52,265; Sulphur Springs, Jefferson Co., Trelease, Oct. 23, 1899, the plant marked A, the fruit all fallen off, Herb. No, 52,266; Winona Lodge, James River, Greene Co., Trelease, July 22, 1897, a fine, full plant just coming into flower, Herb. No. 52,267; Jefferson City, Cole Co., Dr. O. Krause, June, 1866, a fairly complete plant just coming into flower, labeled A. parviflora, Herb. No. 52,268; Williamsville, Wayne Co., T'release, Sept. 9, 1897, a large, complete plant with some fruit still on the branches, Herb. No. 52,269; Allenton, St. Louis Co., Letterman, July 10, 1888, a fine complete plant in flower and fruit, labeled A. 1916] | BUSH—THE MISSOURI AGRIMONIES 315 striata, Herb. No. 52,270; Swope Park, Jackson Co., Mac- kenzie, July 4, 1896, the plant marked B, a fine complete plant just coming into flower, Herb. No. 52,292; Pilot Knob, Iron Co., Glatfelter, Aug. 20, 1895, the plant marked A, a small plant with all the fruit fallen off, labeled A. micro- carpa, Herb. No. 52,292. 3. Agrimonia microcarpa Wallr. Beitr. Bot. 1:39. 1842. Agrimonia pumila Muhl. Cat, 47 (hyponym). 1813; Bick- nell, Bull. T'orr. Bot. Club 23:514. 1896. Only the following have been examined: Terre Bleue Creek, St. Francois Co., Trelease, Aug. 29, 1898, a small plant, 3 dm. tall, with five or six leaves near the base, in young fruit and flower, Herb. No. 52,271. 4. Agrimonia platycarpa Wallr. Beitr. Bot. 11:38. 1842. Agrimonia Eupatoria Gray, Manual, eds. 1-6, in part, and of many other American authors, not L. 1753. The following specimens have been examined: Dodson, Jackson Co., Bush 851, Sept. 2, 1900, a very good, whole plant in ripe fruit, Herb. No. 52,273; Atchison Co., Bush, Aug. 23, 1893, a complete specimen in full fruit, which I refer to this species, Herb. No. 59,319; Cliff Cave, St. Louis Со., 7. В. S. Norton, Aug. 26, 1899, a very poor speci- men with no fruit, but which may well belong to this spe- cies, Herb. No. 52,894; Courtney, Jackson Со., Bush 253, Aug. 21, 1896, a complete specimen in good fruit, which appears to belong to this species, Herb. No. 52,320; Spring- field, Greene Co., Dewart 115, July 31, 1882, one small and one tall slender plant in poor condition, the plant marked B apparently belonging to this species, labeled А. Eupa- toria, Herb. No. 52,316; Hannibal, Marion Co., Davis 6210, Aug. 24, 1915, a full, complete plant in fruit; Swope Park, Jackson Co., Bush, Oct. 14, 1915, full, complete plants in fine fruit. 5. Agrimonia pubescens Wallr. Beitr. Bot. 11:45. 1842. Agrimonia Eupatoria, var. mollis Torr. & Gray, Fl. N. Am. 1:431. 1840. [Vor. 3 316 ANNALS OF THE MISSOURI BOTANICAL GARDEN Agrimoma Eupatoria Gray, Manual, eds, 1-6, in part, and of many other American authors, not L. 1753. Agrimonia parviflora Kinn, in Wallr. Beitr. Bot. 11:45, as synonym. 1842, not A. parviflora Soland. 1789. Agrimoma mollis (T. & G.) Britton, Bull. Torr, Bot. Club 19:221. 1892. The following specimens were examined: Seligman, Barry Co., Dewart, Aug. 21, 1892, a complete plant in flower and fruit, with no specific name, Herb. No. 02,272; Washington Co., Wislieemus 107, July 23, 1885, sev- eral pieces apparently all belonging to one plant, labeled А. Eupatoria, Herb. No. 52,291; Jefferson Barracks, St. Louis Co. Norton, Nov. 17, 1900, a very poor specimen without leaves or fruit, but evidently this species, Herb. No. 52,290; Roaring River, Barry Co., T'release 983, Sept. 7, 1898, a tall plant eut into two pieces, with the fruit all fallen off, Herb. No. 52,296; Flat River, St. Francois Co., Trelease, Oct. 13, 1897, two plants, one very poor, the other with ripe fruit, Herb. No. 52,297; St. Louis, St. Louis Co., Engelmann, Aug., 1842, labeled A. striata, Herb. No. 52,298; Seligman, Barry Co., Dewart, Aug. 21, 1892, a tall plant in fruit, not named, Herb. No. 52,299; Chain of Rocks, St. Louis Co., Craig, Oet. 4, 1908, a poorly pressed, tall plant with nearly all the fruit fallen off, Herb. No. 52,300; Webb City, Jasper Co., Bush 6038, July 23, 1910, a fine, large, com- plete plant just coming into flower, Herb. No. 52,301; Mon- teer, Shannon Co., Bush 6136, Aug. 8, 1910, a tall, fine plant just coming into flower, Herb. No. 52,302; Monteer, Shannon Co., Bush 6135, Aug. 8, 1910, a slender, complete plant in flower and young fruit, Herb. No. 52,303; St. Louis, St. Louis Co., Glatfelter, 1889, the top of a tall, robust plant just coming into flower, labeled A. Eupatoria, Herb, No. 52,304; west of St. Louis, St. Louis Co., Hitchcock, July 26, 1890, a tall, robust plant with all of the fruit fallen off, not named, Herb. No. 52,305; Cass Co., Broadhead, Aug. 11, 1884, two very poor specimens of tops of plants, without flowers or fruit, but evidently this species, Herb. No. 52,306; 1916] BUSH—THE MISSOURI AGRIMONIES 227 Newton Co., Bush, July 15, 1893, a small plant in fruit, labeled A. Eupatoria, Herb. No, 52,307; Greene Co., Blank- inship, Aug. 13, 1889, a slender plant broken in two pieces, in mature fruit, labeled A. microcarpa by me, Herb. No. 52,308; Creve Coeur Lake, St. Louis Co., Norton, Sept. 17, 1898, a poor specimen as to leaves, but with good fruit, Herb. No. 52,309; Kansas City, Jackson Co., Bush 1747, July 29, 1902, a full, complete specimen in flower and fruit, Herb. No. 52,310; Webb City, Jasper Co., Palmer 306, Aug. 30, 1902, a specimen in poor condition, with fruit all fallen off, Herb. No. 52,311; Independence, Jackson Co., Bush 272, Sept. 12, 1895, a poor specimen as to leaves, but with good fruit, Herb. No. 52,312; Monteer, Shannon Co., Bush 745, Aug. 22, 1901, a fine, large plant in good fruit, Herb. No. 52,313; Swan, Taney Co. Bush 721, Oct. 1, 1899, a poor plant as to leaves, but with fine large fruit, Herb, No. 52,314; North Kansas City, Clay Co., Mackenzie, Sept. 12, 1897, a large, fine specimen in fine fruit, Herb. No. 52,315; Spring- field, Greene Co. Dewart 115, July 31, 1892, the plant marked A apparently belonging to this species, Herb. No. 52,316; Bismarck, St. Francois Co., Bush 37, Sept. 12, 1893, the top of a fine plant in fine fruit, Herb. No. 52,317; Me- Donald Co., Bush, Sept. 1, 1893, the top of a tall plant in good condition, but just coming into flower, Herb. No. 53,318; Sulphur Springs, Jefferson Co., T'release, an upper and lower part of a poor plant without fruit, Herb. No. 52,266. 6. Agrimonia parviflora Soland. in Ait. Hort. Kew. 2:130. 1789. Agrimonia Eupatoria Michx. Fl. Bor. Am. 1:287. 1803, not A. Eupatoria L. 1753. Agrimonia suaveolens Pursh. Fl. Am. Sept. 1:336. 18106. Agrimonia serrifolia Wallr. Beitr. Bot. 11:40. 1842. Agrimonia Eupatoria, var. americana Kinn, in Wallr. Beitr. Bot. 1!:40. 1842, as synonym. Agrimonia polyphylla Urban, Symb. Ant. 7:227. 1912. I [Vor. 3 318 ANNALS OF THE MISSOURI BOTANICAL GARDEN Specimens examined: Marble Cave, Stone Co., T'release, Sept. 11, 1898, a large, fine plant in fine fruit, Herb. No. 52,274; Little Blue, Jackson Co., Bush 258, Aug. 9, 1896, a good specimen just coming into flower, Herb. No. 52,275; Texas Co., Blankinship, Aug. 6, 1888, a good specimen just in young fruit, Herb. No. 52,276; St. Louis Co., Eggert, July 27, 1878, a good, full specimen seareely in flower, Herb. No. 52,277; Pilot Knob, Iron Co., Russell, Sept., 1897, a good middle portion of a plant with- out flowers or fruit, Herb. No. 52,278; McDonald Co., Bush, Sept. 1, 1895, a good, full specimen in flower and fruit, Herb. No. 52,279; Jasper Co., Bush, Aug. 16, 1893, a good terminal portion of a large plant in flower and young fruit, Herb. No. 52,280; Pureell, Jasper Co., Palmer 609, Sept. 25, 1904, a weak, undeveloped plant in flower and fruit, Herb. No. 52,281; Monteer, Shannon Co., Push 6134, Aug. 8, 1910, a good, complete specimen just coming into flower, Herb. No. 52,282; Smithfield, Jasper Co., Palmer, Oct. 11, 1908, a good, full specimen with nearly all the fruit fallen off, Herb. No. 52,285; Kimmswick, Jefferson Co., Wislizenus 108, Aug. 23, 1885, a good middle portion of a plant without flowers or fruit, Herb. No. 52,287; Monteer, Shannon Co., Bush 4897, Oct. 10, 1907, a good, complete plant with nearly all the fruit fallen off, Herb. No. 52,283; St. Louis, St. Louis Co., Glatfelter, 1892, a good, complete plant just com- ing into flower, Herb. No. 52,284; Lake City, Jackson Co., Mackenzie, Aug. 9, 1896, a good, complete plant in flower and young fruit, Herb. No. 52,287; Kimmswick, Jefferson Co., Wishzenus 108, Aug. 23, 1885, evidently the upper part of sheet No. 52,289, in good flower and fruit, Herb. No, 52,288. THE THELEPHORACEAE OF NORTH AMERICA. VII? SEPTOBASIDIUM EDWARD ANGUS BURT Mycologist and Librarian to the Missouri Botanical Garden Associate Professor in the Henry Shaw School of Botany of Washington University SEPTOBASIDIUM Septobasidium Patouillard, Jour. de Bot. 6:61. textf. 1892; Essai Taxon. Hym. 7. 1900; Sace. Syll. Fung. 11:118. 1895; 1614. 14:215. 1900; ibid. 16:184. 1902; ibid. 17:203. 1905; ibid. 21:445. 1913. — Jola Moller, A., Bot. Mitth. a. d. Tropfen 8, Protobasidiomyceten 22-29. pl. 4. f. 4. 1895; Engl. & Prantl, Nat. Pflanzenfam. L1**:84. 1897; Sace. Syll. Fung. 14:245. 1900. The genus was founded upon Septobasidium pedicellatum, Pat. and Septobasidium velutinum Pat. Fructifications resupinate, effused, coriaceous, producing probasidia upon the hyphae at or near the hymenial surface ; the probasidia remain attached to the hyphae and either pro- duce at the apex a few-celled, hyaline, spore-bearing filament, or elongate, become septate, and differentiate into such a filament, usually termed a transversely septate basidium; spores simple, hyaline, even, borne one to each cell by the terminal cell and next lower cells. The spores are apparently produced in succession upon the spore-bearing organ rather than simultaneously, for in only two instances have I observed two spores present at the same time upon the same organ; in these the two spores were very unequal in size. One sees a spore attached to the terminal cell more frequently than to lower cells but perhaps Nore.—Explanation in regard to the citation of specimens studied is given in Part VI, Ann. Mo. Bot. Gard. 3:208, footnote. e technical color terms used in this work are those of Ridgway, Color Standards and Nomenclature. Washington, D. C., 1912. 1[ssued November 4, 1916. ANN. Mo. Вот. GARD., Vor. 3, 1916 (319) (Vor. 3 320 ANNALS OF THE MISSOURI BOTANICAL GARDEN because of the more favorable position of the terminal cell. I have frequently observed a spore attached to some one or other of the upper three cells of the spore-bearing organ but have seen such attachment to the fourth cell only in S. cas- taneum, although often noting on the fourth cell in some spe- cies a lateral protuberance similar to those to which spores were attached in the upper cells. The spore-bearing stage is apparently of very brief duration, judging by the few collec- tions which show this stage well. Specimens are usually col- lected sterile or with probasidia. It is hoped that the record given as to the month when each of our species has been col- lected in spore-bearing condition may aid in securing more valuable specimens for study in the future. Septobasidium is not one of the genera of the Thelephora- ceae, for its spore-producing organs are not simple basidia. The genus is treated here merely for the convenience of students of the Thelephoraceae, as in the ease of Tremel- lodendron, Eichleriella, and Sebacina. The coriaceous struc- ture and resupinate habit of the species of Septobasidium are so similar to those of Corticium and other resupinate genera of the T'helephoraceae that examination by the miero- scope of sections of the fructifieation is necessary to distin- guish an unfamiliar species of Septobasidium from Corti- cium, ete. Many of the known species of Septobasidium were originally published as Corticiums and Thelephoras, and it is probable that careful study of authentie specimens of the earlier species of these genera will lead to the transfer of additional species to Septobasidium. It is possible that some authors may have mistaken the pyriform to globose pro- basidia of species of Septobasidium for conidia and have published such species as Hyphomycetes. "The probasidia may be distinguished from hyphomycetous spores by the former bodies remaining attached to the hyphae; the pro- basidia do not float about loose in preparations. I am indebted to Dr. R. P. Burke for transmitting to me in fresh condition spore-bearing material of three species of Septobasidium. Spore falls were obtained from this ma- terial for germination experiments and some material was 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 321 fixed and preserved for a cytological study of Septobasidium during spore production. Discussion of the systematic rela- tionships of Septobasidium may well await the completion of such study. The species of Septobasidium are tropical or subtropical. Extreme northern stations, based on specimens examined by the writer, are London, Ontario, Canada, and Madison, Wisconsin—both are stations for S. pseudopedicellatum, which is the most frequent species of the United States. With regard to the biology of Septobasidium, several speci- mens of this genus—usually of S. pseudopedicellatum—have been noted by their respective collectors as occurring espe- cially on plants badly affected by scale insects. Other speci- mens show scale insects numerous about the fructification and overrun by it. Petch! in a note on the biology of Sep- tobasidium states that from examination of a long series of specimens, it has been determined that these fungi are par- asitie on colonies of scale insects which they overgrow and destroy completely, and that these fungi live, not on secre- tions of the insects, but upon the insects themselves. In addition to independent observations on the association of Septobasidium with scale insects, other facts tending to show an entomogenous adaptation of Septobasidiwm are the following: (1) All species of Septobasidium known to the writer occur only on living branches or leaves, and in no instance has there been penetration by the fungus through the epi- dermis or bark into the living tissues of the substratum, or any injury or deformation or gall response by the branch or leaf. (2) Spores are produced by S. pseudopedicellatum, in the region from North Carolina and Alabama to Porto Rico, in May when young colonies of the scale insects are forming. Mr. Seagle wrote to me that the old fructifications of S. pseudopedicellatum disappear from his apple trees in North Carolina in late spring and in early summer, and new fruc- 'Ann. Bot. 25:843. 1911. [Vor. 3 322 ANNALS OF THE MISSOURI BOTANICAL GARDEN tifications grow which become large by early winter. The collections which I have studied, made during fall and early winter, have been in vegetative rather than in fruiting stage. On the other hand, some specimens of Septobasidium in herbaria have no seals insects on the portions of twigs bear- ing the fructifications of Septobasidium, but I can not say as to whether these fructifications made their start on clean twigs or on scattered scale insects which they have сош- pletely overgrown and destroyed. -- — — — КЕү To THE SPECIES "EP having the hymenial layer or membrane raised above the substratum and supported on scattered pillars composed of parallel h Nahe close together side by в10ө.................................. Fruetifieation having the Cadet zéi hg Ze supported on pillars but with the pillars less regular in for n the above and compo та of loosely interwoven and curving ре eh from Cuba...... . 8. cirratum — laeking supporting hyphal pillars, with 4. Dechen from substratum to the hymenial region without noteworthy eonsoli- dation. TEE *98a49»960»*»94 eee eee eee ee ee ee 2 КУЕ АКТУ ЛУК афа е . With erect or suberect paraphyses or hyphal branches at the surface of the о 554 cee E ICH COQUE SUR ERA OEC CR PCI RUE 2 . With surface of hymenium ce ле = longitudinally arranged and inter- woven paraphyses or hyphal Втапеһев................................. . Structure p^ goes ce of hymenium а pu blished; probasidia 20x15- € kw P a e base of the spore-bearing organs; spore-bearing organ horseshoe- Zeien 56х10. In CUDA 35255%55%%44%4%:40»4»: 8. асе . Structure of surface of hymenium not published ; 5. He shin- hi lou . 8. ing, wer Me, Ted, ex ege NEE EEN a atratum Fruetifieation 25-65 mm. thick; probasidia 127-94; 5 ng Rhett our све іе бө EN ge 2. S. Schweinitats . Fruetifieation 1-1% mm. thick; p 26х11ш; rk 2 4 organs hook-shaped, T to 50x8u. In Mexico ............ S. tropicale Fruetifieation 1-1% mm, thick; о. FC 25х10-13и; spore-bear- ing organs straight, up H 60х11ш; spores 13x5144u. In Jamaica ҚА Gants iE Gard geb ІТ EE 8. „жын 2. EE not shining, velutinous, aniline-blaek, becoming fuse n the herbarium; probasidia 15-206 in diameter ....... . 8. Patouillardii 4 bo bo : ec ПОШ, EE eebe rt Vr ертек ек E EA ааа a КГУ ng 4. Varying from avellaneous and wood-brown to cinnamon-brown; prob а 12-20 8-154; spores 17-22х4-Би ..... .-4. S. ps eudopedicellatum andyke brown when in vase did npe shes: brown ar рарга, Karin 11-15x9-10u; spores 123-3144 p ............ astaneum 4. Olive-brown darkening to ge 2 gray; —1 Geh spor unknown. In Niearagüms ENEE KENE г, еннай s 4 te rw divided into many narrow, sinuous SE Kate shown ара (ПО EE i ee ee ee eme OX OCC a o A RR ч ed iti not divided but with surface reticulated with vi veins at first drab or Prout’s brown then Chaetura-drab........... . 8. rtiforme . Fruetifieation neither divided nor veined .............................. eem ысы. - S E аа ee a ellen, ts, ақа саса. Zéi 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 929 6. pierna when bright colored, often smoke-gray or We mouse-gray, LKE ЗО sss a . Langloisii 6. Honey- кы to old ES velutinous ee a 8. frustulosum 7. Hymenial crust glabrou etween mouse-gray and hair-brown; middle region spongy, lacunose; воп 115-2 mm. thick. In Cuba...... E E ИИ 14. S. Spongia if pU puri акме, between mouse- 2. and hair-brown; poe sidia 12-15 in dia s=spores 19-16хо-би ас sso cre aS rs 15. 8. fumigatum =з RECH E white at fir: Pa pale olive-buff in the herbarium; probasidia 15-17ш in diameter; spores 15-20x5%4-6u. In ROG nia VELAT A RU EE EE . S. canescens 7. Fructification velutinous, between lilac-gray and pallid B e gray; pro- basidia up to 94 in diameter; spores 11-18х8%-4%д. In Trinidad...... REESEN E de s. ЕК 002050: 17. S. lilacinum 1. Septobasidium pedicellatum Patouillard, Jour. de Bot. :61. testi: 1892 Thelephora EE of C. Wright's Cuban Ехвіссай, but not of Schweinitz. Type: in Museum of Paris. Fruetifieation with pillars or pedicels composed of hyphae which branch towards the upper end and pass into and sup- port the hymenial crust; probasidia subglobose, 20 X 15-20y, arising as lateral outgrowths near the ends of the final branches of the hyphae, producing from the apex a hyaline, cylindric, spore-bearing organ, 35 X 104, 2-3-septate, which becomes horseshoe-shaped, slightly constricted at the septa, and has a small protruberance on the convex side of each cell; no spores seen. The above is a summary of the account by Patouillard,! of the structure of the specimen in the Museum of Paris, eolleeted in Cuba by C. Wright and distributed by him in his Cuban exsiecati under the name Thelephora pedicellata. Wright made two collections in Cuba which were determined by Berkeley and Curtis? as Thelephora pedicellata. Since Patouillard omitted the data on the label of the specimen whieh he studied, I do not know now which of Wright's numbers is the type collection and have to defer a fuller consideration of this species to the supplement to my mono- graph. с Zoe, cit. 3Linn. бос. Bot. Jour. 10:329. 1868. [Vor. 3 324 ANNALS OF THE MISSOURI BOTANICAL GARDEN 2. S. Schweinitzii Burt, n. sp. Thelephora pedicellata Schweinitz, Naturforsch. Ges. Leip- zig Schrift. 1:108. pl. 2. f. 3. 1822; Fries, Elenchus Fung. 1:200. 1828; Epicr. 544. 1838; Sace. Syll. Fung. 6:544. 1888. Not Septobasidium pedicellatum Pat. Illustrations: Schweinitz, loc. cit. Type: in Herb. Schweinitz. Fructification resupinate, coriaceous, dry, not separable from the substratum, varying from drab and cinnamon-drab to wood-brown, the margin undulate, whitish; in structure 3-layered, with (1) a layer next to the substratum of densely interwoven, colored hyphae 3—31⁄ in diameter, which form (2) a layer of erect hyphal pillars or pedi- * eels each about 200-3004 long, 40-754 | in diameter, about 2 to a millimeter, and pass into and support at the outer end (3) the hymenial layer 120-200 thick, composed of densely inter- x а woven, colored hyphae 3-314. in di- a, portion of hymeni. ameter, of erect, flexuous, filiform, DNA š sparingly branched, hyaline paraphy- spore-bearing organ and ses or hyphal branches about 110и in IM мы a X diameter, and, when in fertile stage, of hyaline, thin-walled, erect probasi- dia, pyriform to subglobose, 12 X (-Әшіп the type, borne on the colored hyphae; spores simple, hyaline, even, curved, 5 X 2154 (as seen attached in the type but perhaps immature), borne singly at the apex of the terminal cell of a short filament 20-95 X 41-би, about 4 cells long, eurved to fish-hook-shaped in form, which de- velops from the probasidium. Fructifications 2 — several em. long, 1 — several em. broad, 12-12 mm. thick. On living branches. North Carolina to Louisiana. Fre- quent in winter. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 325 The above description is based on the Schweinitzian type and presents the characters of a rare species which has not been distinguished heretofore from the following S. pseudo- pedicellatum, a thicker, larger, common, and widely distri- buted species. S. Schweinitzii is characterized by its erect filiform paraphyses, curved to hook-shaped, spore-bearing organs, and small spores, although it is not certain that full- sized mature spores have yet been seen. I refer to S. Schweinitzii a collection made by P. L. Ricker on Persea, in Georgia, during August, because this specimen has small probasidia, hook-shaped, few-celled, hyaline, spore- bearing organs, and spores 7 X Alan: but in this specimen only a few paraphyses are present, the probasidia and hook- shaped organs are at the very surface of the hymenium, and small, globose organs би in diameter are occasionally pres- ent, borne laterally on the hyphae in the lower part of the hymenial layer. I have not studied with the microscope the Cuban specimen of S. pedicellatum, collected by C. Wright, one of the species upon the structure of which Patouillard founded the genus Septobasidium. He found this specimen to have probasidia and hook-shaped organs. Both proba- sidia and the hyaline organs are described as larger than they measure in the Sehweinitzian type. In the Cuban speci- men the probasidia are stated to be 20& in diameter or 20 x 15и, and the hook-shaped organs as 35 X 10и, and the former persist full size, with the septate hook-shaped organs connected with them like a promycelium with its teleutospore. These differences indicate that the Cuban specimen belongs to a species distinct from Thelephora pedicellata Schw. It is necessary to substitute a new specific name for '*pedicellata"' in making the transfer of Thelephora pedicellata Schw. to Septobasidium, because there is already a valid Septobasi- dium pedicellatum. Specimens examined: North Carolina: Schweinitz, type (in Herb. Schw.). Georgia: Bugaboo Island, Okeefenokee Swamp, P. L. Ricker, 921. я [Yor, 8 326 ANNALS OF THE MISSOURI BOTANICAL GARDEN Louisiana: Gibson, F. T. McLean, comm. by P. Spaulding. 3. S.tropicale Burt, n. sp. Type: in Mo. Bot. Gard. Herb. and in Farlow Herb. Fruetifieation resupinate, effused, coriaceous, dry, not sep- arable from substratum, glabrous, not shining, avellaneous, the margin concolorous, squamulose-fimbriate, not closely ad- nate; in structure 3-layered, with (1) a layer next to the substratum of dense- ly interwoven, concolorous, thick-walled hyphae 3-314. in diameter, which pass into and form (2) a layer of nu- merous erect, slender pillars about 40и in diameter, 5 or 6 to the millimeter, whose hyphae spread apart at the outer end, branch, and form 8. cale. a, probasidium ; T io spore- пра and support (3) the hyme- EE а и nial crust about 200, thick, densely interwoven through- out, with the even, thick-walled, colored hyphae up to би in diameter on the under side, more erect, paler, and about 2p in diameter at the surface; probasidia terminal on the hy- phae, hyaline or but slightly colored, pyriform, 26 X Пр, at the surface of the hymenium; a spore partially imbedded in the hymenium is hyaline, simple, even, curved, 19 X би, no others seen; fish-hook-shaped organs, such аз probably bear the spores, are present in the surfaee of the hymenium, several-celled, up to 50 X 8», with prominent protuberances from cells on the convex side of the organ. Fructification 4 em. long, about 2 em. broad, 1-115 mm. thick. On bark of living branches of Quercus. Mexico. The distinctive characters of this species are avellaneous color, surface not shining, margin squamulose-fimbriate, not closely adnate as in the preceding species, and thicker hy- 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 327 menial crust not loosely interwoven on its under sides, pro- basidia terminal on the hyphae, and the large hook-shaped, presumably spore-bearing, organs of the upper surface. If these organs grow out from the probasidia, the probasidium must differentiate into the organ, for I have traced the eurved organ back to the colored hyphal cells. Specimens examined: Mexico: locality not stated, C. G. Pringle, comm. by W. G. Farlow, 5 (in Mo. Bot. Gard. Herb., 44590). 4. S. pseudopedicellatum Burt, n. sp. Thelephora pedicellata of most American authors but not of Schweinitz. Type: in Mo. Bot. Gard. Herb. Fructification resupinate, effused, coriaceous, dry, not sep- arable from the substratum, varying from avellaneous and = ЖЖ Quin к | Fig. 3 S. pseudopedicellatum. h, portion of hymenium showing the longitudinally inter- woven hyphal ends or paraphyses and some probasidia; b, three spore-bearing organs; s, spores. x 640. wood-brown to cinnamon-brown, the margin undulate, whit- ish; in structure three-layered, with (1) a layer next to the substratum of densely interwoven, thick-walled, slightly col- ored hyphae 3» in diameter, which form (2) a layer of erect, hyphal pillars, or pedicels, each about 500» long, 20- 40 in diameter, about 3-5 to a millimeter, whose hyphae spread apart at the upper end of the pillars, branch, and form and support (3) the hymenial crust about 300, thick, with hyphae loosely interwoven near the pillars, 3-314 in [VoL. 3 328 ANNALS OF THE MISSOURI BOTANICAL GARDEN diameter, very dense at the outer surface with the hyphal branches or paraphyses 2» in diameter, curved longitudinally along the surface and densely interwoven; erect probasidia nearly hyaline, rich in protoplasm, deeply iE о, pyriform, 12-20 X 8-155, are borne laterally on the hyphae about 15» below the surface of the hymenium; spores white in a spore collection, simple, even, curved, 17-22 X 4-би, are borne singly from each of the upper three cells (so far as observed) of a straight or flexuous, few-celled, hyaline organ up to 60 X 5-5%а, which grows from the probasidium and pro- trudes above the surface of the hymenium. Fructifications 2-15 em. long, 1-8 em. broad, 1-115 mm. thick. On small, living branches of apple, orange, oak, Nyssa, Cornus, acu ere and also on orange leaves in one col- lection; sometimes, perhaps always, associated with scale inel. Canada to Florida and Louisiana and westward to Wisconsin; also in Cuba and Porto Rico. December to Au- gust; spores produced in the last of May. S. pseudopedicellatum is the common Septobasidium of southeastern United States. It may be recognized by its brown, glabrous, shining, foliaceous crust which is raised and supported about a millimeter above the substratum on perpendicular, hyphal pillars which are as conspicuous as the rhizoids of a lichen. Old specimens may crack, break the hyphal pillars, and the hymenial crust curl outward so as to show the broken pillars attached to the under side. Sterile specimens of this species have been heretofore referred to S. pedicellatum, but a collection of fertile specimens received from Dr. R. P. Burke in May of the present year shows that our common species differs from S. pedicellatum by having large spores produeed on a straight or but slightly curved, mueh larger, spore-bearing organ, paraphyses or hyphal branches at the surface of the hymenium curved and densely longitudinally interwoven, larger probasidia, and larger and thieker fruetifieations. Even in sterile condition the longitu- dinally interwoven paraphyses are suffieiently distinetive. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 329 Specimens examined: Exsiceati: Ellis, N. Am. Fungi, 12, under the name Thele- phora pedicellata. Canada: Ontario, London, 7. Dearness, 3396 (in Mo. Bot. Gard. Herb., 43802). New Jersey: Newfield, J. B. Ellis; also from same locality in Ellis, N. Am. Fungi, 12. Pennsylvania: Trexlertown, W. Herbst, comm. by Lloyd Herb., 2232. North Carolina: Reepsville, J. P. Seagle, two collections, one of which was communicated by F. L. Stevens. Florida: W. W. Calkins; Daytona, R. Thaxter, 75a (in Far- low Herb. and in Mo. Bot. Gard. Herb., 43894); Kissim- mee, comm. by F. C. Wolf (in Mo. Bot. Gard. Herb., 44205); same locality, D. E. Evans (in Mo. Bot. Gard. Herb. 44403); Ft. Myers, H. S. Fawcett (in Fawcett Herb.); Gainesville, H. E. Stevens, comm. by E. Bartholo- mew, 40b (in Mo. Bot. Gard. Herb., 44212). Alabama: Peters, 75 (in Curtis Herb.); Е. S. Earle Ф C. F. Baker (in Lloyd Herb., 3454); Auburn, Alabama Bio- logical Survey; Montgomery, R. P. Burke, 49, and the type collection (in Mo. Bot. Gard. Herb., 10979, and 20659, type). Louisiana: Gibson, F. T. McLean, comm. by P. Spaulding; St. Martinville, A. B. Langlois, three collections, two of which are (in Lloyd Herb., 2411, 3533). Kentucky: comm. by A. H. Gilbert (in Mo. Bot. Gard. Herb., 44323); ‘‘in mountains,” P. Garman (in Mo. Bot. Gard. Herb., 44302). Wisconsin: Madison, W. Trelease (in Mo. Bot. Gard. Herb., 5164). Cuba: Ceballos, H. S. Fawcett, 10, 39 (in Mo. Bot. Gard. Herb., 15005, 15018); Isle of Pines, H. S. Fawcett, 15 (in Mo. Bot. Gard. Herb., 15094). Porto Rico: Mayaguez, F. S. Earle, 79, N. Y. Bot. Gard. erb. [Vor. 3 330 ANNALS OF THE MISSOURI BOTANICAL GARDEN 5. S. castaneum Burt, n. sp. Type: in Mo. Bot. Gard. Herb. Fructification resupinate, effused, coriaceous, dry, not sep- arable from the substratum, glabrous, cracking in drying into pieces about 10 X 5 mm., olive-brown when fertile, Van- dyke brown when in vegetative condition, the margin соп- colorous; in structure 3-layered, with (1) a layer next to substratum of opaque, concolorous hyphae 4и in diameter, which form (2) a layer of pillared or spongy structure, in some places with pillars up to 150 in diameter, about 1 mm. apart, and in other places with a spongy mass of obliquely ascending, interwoven hyphae similar to those of the pillars. This layer supports (3) the hymenial crust, sometimes (/ \ stratose, with hyphae loosely As interwoven on the under pose side, 3—44 in diameter, very dense at the outer surface, at + ft ` with the hyphal branches or MJ a paraphyses 2и in diameter, >J curved longitudinally along Fig. 4 the surfaee and densely in- terwoven; erect probasidia base of the spore-bearing organ which slightly colored, rich in pro- жен gest 3 = en ne toplasm, deeply staining, pyriform, 11-15 >x 9-10y, are borne laterally on the hyphae about 15и below the surface of the hymenium; spores hyaline, simple, even, curved, 12 X 3-314, borne singly from each of the upper four cells of a straight, few-celled, even-walled, clavate, hyaline organ 30-40 X би, which grows from the probasidium and protrudes above the surface of the hymenium. Fructification 8-15 em. long, wholly surrounding limbs 215 em. in diameter, 1-114 mm. thick. On living bark in swamp, Montgomery, Alabama. May and August—fertile in May. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 523 This species is closely related to 8. pseudopedicellatum but is more deeply colored, has more opaque hyphae, and smaller spores and spore-bearing organs. No lateral pro- tuberances or papillae have been observed on the latter. Specimens examined: Alabama: Montgomery, R. P. Burke, two collections (in Mo. Bot. Gard. Herb., 20421, type, and 20693). 6. S.sublilacinum (Ellis & Ev.) Burt, n. comb. Thelephora sublilacina Ellis & Ev. State Univ. Iowa, Lab. Nat. Sel. Bul. 13:67. 1896; Sace. Syll. Fung. 14:214. 1900. Type: in N. Y. Bot. Gard. Herb. Fruetifieation resupinate, effused, coriaceous, dry, not sep- arable from the substratum, glabrous, shining, olive-brown, darkening to dark neutral gray; in structure 3-layered, with (1) a layer next to the substratum, 40-60» thick, of closely crowded, longitudinally arranged hyphae concolorous with the fructification, 4-414, in diameter, which form (2) a layer of pillars 40-60» in diameter, about 2-4 to a millimeter, whose hyphae spread apart at the outer end and form and support (3) the hymenial crust about 60. thick, densely interwoven throughout, with even, thick-walled, concolorous hyphae 3- 314» in diameter on the under side, Ән in diameter, nearly hyaline, and densely, longitudinally interwoven at the sur- face; probasidia, spores or other organs not present in the type. Fruetifieation about 14 em. in diameter, ?4 mm. thick. On living branches. Nicaragua. The type specimen of this species, when viewed from above, agrees so closely with Ше cotype of S. Spongia in color and habit that one is strongly disposed to regard the two speci- mens as of the same species. S. sublilacinum has, however, the coarser hyphae, a three-layered structure, and distinct pillars. It seems best to regard it as a distinct species, at least until fertile specimens define the species more definitely. Specimens examined: Nicaragua: С. L. Smith, 108, type (in N. Y. Bot. Gard. Herb.). [Vor. 3 592 ANNALS OF THE MISSOURI BOTANICAL GARDEN 7. S. Patouillardii Burt, n. sp. S. (very near) Leprieuri? (Mont.) Patouillard, бос. Myc. Fr. Bul. 16:55. 1900. Type: in Burt Herb. Fruetifieation resupinate, effused, coriaceous, dry, velu- tinous, aniline-black at first, becoming fuscous in the her- barium, the margin rather thick and determinate; in struc- ture 200-400» thick, with (1) next to the substratum a thin layer of loosely inter- woven hyphae За in diameter, buffy brown | | under the microscope, which form (2) а layer of hyphal pillars each about 30-50 in diameter, 100-200» long, about 3-4 to a + "- millimeter, whose hyphae spread apart 4 |! above and form (3) the interwoven hyme- Fig. 5 nial layer containing some probasidia and lard PUT rer H with the surfaee eomposed of numerous spore-bearing orga erect, nearly straight, fuscous hyphal p, four paraphyses or | l * . ° hyphal ends. x 640. branches or paraphyses 2а in diameter; probasidia hyaline, subglobose, 15-20» in diameter, erect on short branches of the colored hyphae; no spores found; the only possible spore-bearing organ seen is 46 X 714, acuminate at the apex. Fructifications 2-315 em. long, 1-2 em. broad, 200-400 mm. thick. On living branches of ash, Liquidambar, and Nyssa. Flor- ida to Louisiana. November to March; a January collection has a few probasidia. This species may be recognized by its thin fructification resembling a piece of black velvet, slightly raised from the hcm on such short and slender pillars as to be barely visible without the aid of a lens. Patouillard determined this species for Mr. Langlois as very near to S. Leprieurit. Since Corticium Leprieurii was originally described as gla- brous, shining, and chocolate-colored, and since no specimens like ours have yet been collected in the region between Guiana and the United States, our specimens are probably a distinct species which should have a definite name. 1916] BURT—TH CEAE OF NORTH AMERICA. VII 3359 Specimens examined: Florida: Daytona, R. Thaxter, 75b (in Farlow Herb. and Mo. Bot. Gard. Herb., 45895). Alabama: Auburn, F. S. Earle Ф С. Е. Baker, also (in Mo. Bot. Gard. Herb., 5165). Louisiana: St. Martinville, 4. В. Langlois, 3005, determined by Patouillard as S. (very near) Leprieurti; Gibson, Е. T. McLean, comm. by P. Spaulding, type—some fragments near a specimen of another species, but having probasidia, etc., as drawn, taken as the type because more mature than other collections cited. 8. S. jamaicaense Durt, n. sp. Type: in Burt Herb. and N. Y. Bot. Gard. Herb. Fruetifieation resupinate, effused, coriaceous, spongy, dry, thiek, bister, with the subieulum bone-brown; in structure with (1) next to the substratum a thin layer of interwoven hyphae A which form (2) a layer of probably oblique, weak, very slender, crowded, hyphal pillars 12-20» in 2 diameter, up to 20004 long, with hy- phae even, 4-би in diameter, buffy brown under the microscope, di- a % verging above to form (3) a spongy hymenial layer 300-400и thick, with hyphae which rise obliquely, are Fig. 6 4 S. jamaieaense. loosely interwoven, and bear proba- һ, hyphal end bearing two sidia laterally at the outer surface probasidia in the surface of the " 2 enium; а, probasidium; b, of the layer and terminate in hya- sporebearing organ; s, spore. line or subhyaline, curved branches х8 or tips; probasidia hyaline, subglobose or pyriform, 13-25 X 10-134, quickly developing into hyaline, straight, few-celled, spore-bearing organs up to 60 X 11и; spores simple, hyaline, slightly curved, 13 X 51%и. Fructification larger than 6 cm. long, 2 em. broad, 1-114 mm. thiek—fraetured on all sides and not showing natural margin. [VoL. 3 334 ANNALS OF THE MISSOURI BOTANICAL GARDEN On bark. Base of John Crow Peak, altitude 5500 feet. Jamaica. April. The type of this species has so thick and spongy a hyme- nial layer that I have tried to regard this specimen as the fertile stage of S. Spongia, but the well-developed layer of pillars is in the way of such reference and the hyphae are rather coarser than in S. Spongia. Specimens examined: Jamaica: John Crow Peak, L. M. Underwood, 2439. 9. S.atratum Patouillard, Soc. Мус. Fr. Bul. 16:181. 1900. Type: location unknown. Fructification resupinate, greatly extended, glabrous, shin- ing, thin, with the margin fimbriate and inerusting; subicu- lum black, formed of rigid, erect, short bundles composed of hyphae but little branched, 4-5 in diameter, with the wall thiek and brown under the mieroscope; hymenial erust thin, fragile, continuous, glabrous, ombre noir, paler at the per- iphery; probasidia at first globose, 10-124 in diameter, growing on the sides of erect hyphae of the hymenial crust a little below their ends; spores and spore-bearing organs not present. On living trunk of Eugenia Jambos. Morne Gommier, near Galion, Guadeloupe. P. Duss. E In connection with the original description, Patouillard stated that S. Spongia is **epais, roux, spongieux, lacuneux,’’ and that S. atratum is ‘‘tres mince, et noir." I have seen no specimens of S. atratwm and base the above account of this species wholly on the original description. 10. S.cirratum Burt, n. sp. Type: in Mo. Bot. Gard. Herb. and Humphrey Herb. Fruetifieation resupinate, effused, coriaceous, spongy, dry, cracked, velutinous, between Benzo-brown and brownish drab, with fuscous subiculum, the margin divided into nar- row, sinuous divisions; in structure up to 7004 thick, with (1) next to the substratum a layer of interwoven hyphae, whieh form (2) a layer of pillars not uniform in diameter, composed of hyphae loosely interwoven, curled together, 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 335 suggestive of ringlets in sectional preparations, which sup- port (3) the hymenial layer 200-3004 thick, with hyphae 2-214» in diameter, notably curved, branched, and loosely interwoven, olive-brown under the microscope, bearing in the lower part of the layer numerous concolorous, globose bodies Пи in diameter, and toward the outer surface hyaline probasidia Па in diameter also, and termi- nating at the surface in fine, hya- line branches Ту in diameter, with recurved or coiled tips; spores from deeper portion of hymenial simple, hyaline, even, curved, 18 layer; > SE paraphyses; s, ў spore. X би; spore-bearing organs few- celled, straight, cylindric, about 35-40 X Тов, differentiating from the probasidia. Fructifications 5 em. long, 114 em. broad. On trunk of living hardwood tree near the base. Cuba. December. Seen but once by the collector. S. cirratum has so nearly the color and habit of Hypochnus fuscus that it was a surprise to find the specimen a Septo- basidium. The color and sinuously divided margin suggest S. Langlois. The pillars composed of loosely interwoven and curving hyphae are unique and separate this species sharply from all our species of the S. pedicellatum group. The hyphae are too fine and too curving for S. Spongia. Specimens examined: Cuba: Omaja, C. J. Humphrey, 2773 (in Mo. Bot. Gard. Herb., 15836). 11. S. Langloisii Patouillard, бос. Мус. Fr. Bul. 16:54. 1900. Type: a portion in Burt Herb. Fructification resupinate, effused, dry, velutinous, plum- beous when bright colored, but often smoke-gray or pallid mouse-gray, repeatedly divided into many narrow, sinuous [Vor. 3 336 ANNALS OF THE MISSOURI BOTANICAL GARDEN divisions which are more distinct towards the margin; in structure 200-250, thick, with hyphae fuscous under the micro- scope, thick-walled, even, loosely interwoven from substratum to hymenium, densely interwoven in the hymenium and bearing hya- line, flexuous, suberect terminal + branches or paraphyses and hya- line probasidia which are ех- Fig. 8 ceeded by the paraphyses; spores Langloisi e . А , probasidium ins a hyaline, simple, even, slightly проте beant oim] py pare. curved, 15-21 X 5-71, appar- physes; s, spores. x 640. ently produced singly at the apex of a nearly straight, 2-3-celled, spore-bearing organ into which the probasidium develops. Fructification up to 5 em. long, 215 em. broad, 1⁄4 mm. thick. On bark of living branches of Crataegus, Carpinus, and water oak. Florida to Louisiana and in Grenada. Novem- ber to May. This species resembles 9. frustuloswm in having the fruc- tifieation divided into narrow sinuous divisions and differs from that species in being blue colored, verging into smoke- gray or paler in some specimens, instead of honey-yellow. The speeimen from Grenada is thinner than those from other localities. Specimens examined: Ехвіссай: Ravenel, Fungi Am., 450, under the name Ste- reum pruinatum. Florida: Gainesville, H. E. Stevens, comm. by E. Bartholo- mew, 40a (in Mo. Bot. Gard. Herb., 44211); same locality, Ravenel, in Ravenel, Fungi Am., 450. Alabama: Montgomery, R. P. Burke, 52 (in Mo. Bot. Gard. Herb., 9558). Louisiana: St. Martinville, 4. B. Langlois, 2995, type. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA, VII Si Grenada: Grand Etang, R. Thazter, comm. by W. G. Far- low, a (in Mo. Bot. Gard. Herb., 43912). 12. 8. frustulosum (Berk. & Curtis) Patouillard, бос. Myc. Fr. Bul. 10:79. pl. 3. f. 4. 1894. Hymenochaete frustulosa Berk. & Curtis, Linn. Soc. Bot. Jour. 10:334. 1868; Басс. Syll. Fung. 6:601. 1888. Illustrations: Patouillard, loc. cit. Type: type and cotype in Kew Herb. and Curtis Herb. Fruetifieation resupinate, effused, coriaceous, dry, velutin- ous, honey-yellow to old gold, repeatedly divided into many narrow, sinuous, reticulate di- visions which are more dis- tinct towards the margin; in structure about 600-700» thick, 3-layered, with next to the substratum a broad layer, up to 200» thick, with hyphae densely longitudinally ar- ranged, 2и in diameter, concol- j orous with the fructification, Fig. 9 which ascend, without forming — , ee Ke pillars, as (2) the loosely ar- ing organs; n, septate colored or- ranged middle layer, whose hy- Saring a spore Чек m oper то ат phae pass into and form (3) the hymenial crust which is finally very dense and compact in fully developed specimens, about 200 thick, with hyphae concolorous, even, 1144-2» in diameter, branching towards the surface into flexuous branches, or paraphyses, about Тр in diameter, once or twice dichotomously branched and with tips curved or spirally coiled; probasidia borne laterally on the hyphae, hyaline, pyriform, 9 X 514, becoming elongated and septate as a few-celled, spore-bearing organ, or producing directly a sterigma bearing one spore; spores hyaline, even, cylindric, nearly straight, 13-17 x 4-би. Fruetifieations up to 10 em. long, 1-2 em. broad, less than 1 mm. thick. 22 [Vor. 3 338 ANNALS OF THE MISSOURI BOTANICAL GARDEN On bark of living limbs of frondose species. Mexico, West Indies, and Venezuela. February, March, November; spore- bearing in November. This species is highly distinguished by honey-yellow color and the division of its fructification into narrow, sinuous, branched divisions, resembling those of the thallus of the lichens, Physcia stellaris and P. obscura. Spore-bear- ing organs are not abundant in the only fertile specimen which I have seen. They appear to become somewhat cork- screw-shaped, with no indication of bearing spores except on the terminal cell, but I was not certain on this point because the occasional attached spores were along the edge of thick sections where only the apex of the organ extended beyond the paraphyses. In two cases probasidia were bearing at the apex, each a body of the form and dimensions of a spore of this species. In the deeper portions of the fructifications brown, pyriform bodies of the same size and form as the probasidia are borne by the hyphae in the same location as the probasidia. These brown organs are often of the same dimensions as the spore-bearing organs, septate, and gorged with brown contents. Specimens examined: Exsiceati: Smith, Cent. Am. Fungi, 100, under the name Thelephora retiformas. Mexico: Sanborn, Oaxaca, C. R. Orcutt, 3334 (in Mo. Bot. Gard. Herb.). Nicaragua: Castillo Viejo, C. L. Smith, in Smith, Cent. Am. Fungi, 100. Cuba: C. Wright, 244, cotype (in Curtis Herb.). Grenada: Grand tang, R. Thaxter, comm. by W. С. Far- low, 11. Venezuela: Fendler, 279 (in Farlow Herb. and in Mo. Bot. Gard. Herb., 20411). 13. S. retiforme (Berk. & Curtis) Patouillard, Soc. Mye. Fr. Bul. 16: 55. 1900. Thelephora retiformis Berk. & Curtis, Linn. Soc. Bot. Jour. 10:330. 1868; Sace. Syll. Fung. 6:544. 1888. 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 339 Type: type and cotype in Kew Herb. and Curtis Herb. Fruetification resupinate, effused, coriaceous, at first drab or Prout’s brown, then Chaetura-drab, the hymenial surface reticulated with obtuse veins, pulverulent; in structure 700, thick, with the hyphae colored, 3-4» in diameter, short-celled, loosely interwoven or rising obliquely from substratum to hymenial sur- face and there densely interwoven longitudinally and bearing laterally brown, globose or pyriform bodies 13-15 X 10-13», and slightly colored proba- Fig. 10 sidia of the same size and form; a single spore in S. retiforme. the hymenial surface is hyaline, even, curved, 15 dium; n, col- X 4н; no spore-bearing organs found. 8, Spore. x Fructification 1-4 em. long, about 700и thick. 649, On living branches of apple, pear, peach, Carya. District of Columbia to Louisiana and Cuba. November to February, producing probasidia in February. S. retiforme resembles a small foliaceous lichen in habit. It may be distinguished from our other species by its drab to brown color and reticulately veined hymenial surface. The spore characters stated are uncertain for only one spore was seen. Specimens examined: Exsiecati: Ellis & Ev., N. Am. Fungi, 2604. Distriet of Columbia: Washington, comm. by Mrs. F. W. Patterson. Georgia: Fort Valley, comm. by Mrs. F. W. Patterson. Alabama: Forestdale, C. C. Woodward, comm. by J. B. Rorer; Abbeville, S. T. Slaton (also in Lloyd Herb., 3460, and in Mo. Bot. Gard. Herb., 5166). Louisiana: St. Martinville, 4. В. Langlois, 2233. Cuba: C. Wright, 288, cotype (in Curtis Herb.). 14. 6. a MON & Curtis) Patouillard, Soc. Myc. Fr. Bul. 16:181. Thelephora Tae Berk. & Curtis, Linn. Soc. Bot. Jour. 10:330. 1868; басе. Syll. Fung. 6:542. 1888. Type: type and eotype in Kew Herb. and Curtis Herb. [Vor, 3 340 ANNALS OF THE MISSOURI BOTANICAL GARDEN Fruetifieation resupinate, effused, not separable from the substratum, dry, glabrous, shining, between mouse-gray and hair-brown, the margin strigose; in structure lacunose, spongy, about 1 mm. thick when dry, distending SMS to 115-2 mm. when moistened, with hyphae ES 556) 247 3-5 м in diameter, fuscous to clove-brown un- Fig. Я der the microscope, densely longitudinally ar- S. Spongia ranged in masses along the substratum and ris- ае Mieri ing obliquely so as to form a spongy structure fication show- With vacant spaces up to 800 X 500, united iag abre sg, above into a continuous hymenial crust 40-804 thick; probasidia, spores, and spore-bearing organs not present. Fructification ‘‘spreading for many inches," 115-2 mm. thick. On bark of cacao trees. Cuba. S. Spongia is distinguished from our other species in the group having a glabrous hymenial erust by the spongy, rather than pillared, strueture of the middle region of the fruetifieation. The surface of the cotype is infested with a colorless hyphomyeete whose hyphae are densely crowded together and agglutinated; hence fertile specimens of this species will probably be browner than the original sterile, infested specimen. Specimens examined: Cuba: C. Wright, 566, сойуре (in Curtis Herb., and a por- tion in Mo. Bot. Gard. Herb., 44592, by kindness of Dr. Farlow). ? Mexieo: Monterey, Sierra Madre, C. G. Pringle, comm. by W. G. Farlow, 6 (in Mo. Bot. Gard. Herb., 44591). 15. S.fumigatum Burt, n. sp. Type: in Mo. Bot. Gard. Herb. and Humphrey Herb. Fructification resupinate, long and broadly effused, not separable from substratum, coriaceous, tomentose, between mouse-gray and hair-brown, rarely with surface pale, the margin thinning out and concolorous; in structure 800-1500, thick, with hyphae buffy brown under the microscope, even, thick-walled, An in diameter, loosely interwoven and ascend- Te ECHT a 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 341 ing from substratum to hymenial region; in the hymenial region the hyphae become more densely interwoven and bear laterally numerous hya- line, subglobose probasidia 12-154 in diameter, and terminate in small, curved (Y or loosely coiled, colored S branches—hardly paraphy- ( Тай „2— Р ^ . E ses—2x in diameter, which < ЗА Íorm Ше surface of Ше hy- N \ menium; spores simple, hya- | line, even, slightly curved, Un, Sr 2 12-15 x 5-би, borne on Ше j x: Fig. 12 upper three cells of a few- . 8. fumigatum. celled, nearly straight, hya- h, portion of hymenium showing hy- line, spore-bearing organ 40- phae bearing probasidia and terminat- Š 2 Б ing in curved and coiled ends; b, three 50 x 6-7 1и, into which Ше spore-bearing organs; s, spores. X 640. probasidium develops. Up to 5 m. long, several em. broad, 45-14 mm. thick. On trunks of living sapling of Acer rubrum and probably other species. South Carolina, Alabama, and Cuba. Novem- ber to May; spores most numerous in May. S. fumigatum has the general habit and color of Hypoch- nus spongiosus and is readily distinguishable among the North American species of Septobasidium by its mouse- gray color, tomentose surface, and felty structure of loosely interwoven hyphae which do not form pillars. It is only rarely that I have seen spores or evidences of spore produc- tion upon other than the terminal cell in this species. Specimens examined: South Carolina: Gourdin, C. J. Humphrey, 2588, type (in Mo. Bot. Gard. Herb., 48822). Alabama: Montgomery, R. P. Burke, 50, and an wnnumbered collection (in Mo. Bot. Gard. Herb., 1138°, 20068). Cuba: C. Wright, Fungi Cubenses Wrightiani, 838, comm. by W. G. Farlow (in Farlow Herb. and Mo. Bot. Gard. Herb., 43907). [Vor. 3 342 ANNALS OF THE MISSOURI BOTANICAL GARDEN 16. S. canescens Burt, n. sp. Type: in Mo, Bot. Gard. Herb. Fruetifieation resupinate, effused, coriaceous, cottony, pubescent, white 5 at first, pale olive-buff in Ше herbar- ¢ Г ium, the margin thinning out; in " | structure 500-900, thick, with the hy- -2 " phae hyaline, even, 414» in diameter, densely interwoven next to the sub- "d stratum, suberect, or ascending p, obliquely and loosely interwoven to f) Ы | the hymenial surface and Шеге bear- E 4 Ñ ing probasidia laterally among slight- Fig. 13 ly curved hyphal branches about 2» in Ae серна ИЕ diameter; probasidia hyaline, subglo- roo banag med bose, 15-17» in diameter, producing а ia; р, two spore-bearing | few-celled organ 7154 in diameter, up organs; s, spores. X I š 640. to 60u long, which bears spores on its upper three cells; spores simple, hyaline, even, curved, 15-20 X ощ-би. Fructifications about 2-4 em. long, 1-114 em. broad, some- times arranged more or less interruptedly for up to 25 em. along the under side of limbs. Associated fairly constantly with scale insects on small living branches of Quercus on a residence street, Pasadena, California. November to March. 5. canescens is characterized by its white to whitish color, cottony structure, and pubescent surface. Spores were ob- served attached to one or more of the upper three cells of the spore-bearing organ but with the terminal cell giving the most indication of spore production. I am indebted to Prof. H. S. Faweett for the collection made in March to show this species in best fruiting condition. Specimens examined: California: Pasadena, H. S. Fawcett, comm. by W. A. Setehell (in Mo. Bot. Gard. Herb., 44037); same locality, А. G. Smith, comm. by H. S. Fawcett (in Mo. Bot. Gard. Herb., 44246). 1916] BURT—THELEPHORACEAE OF NORTH AMERICA. VII 343 17. S. lilacinum Burt, n. sp. Type: in Farlow Herb. and Burt Herb. Fruetifieation resupinate, effused, coriaceous, dry, adnate, velutinous, between lilae-gray and pallid smoke-gray, the margin adnate, fimbriate; in structure 80-200» thick, with hyphae thin-walled, 22 2-214 in diameter, somewhat colored S near the substratum, ascendine and in- terwoven and becoming hyaline towards the hymenium, finer, 114» in diameter, арт and bearing laterally probasidia, and a \ extending beyond the probasidia and Fig. 14 branching, with tips curved to form the velvety surface of the hymenium; proba- hypha in hymenial sur- АР А 4 А face bearing а probasidi- sidia hyaline, even, globose, up to 94 in um; a, mature probasidi- diameter; spores simple, hyaline, even, съ ДЫ dar cr slightly curved, 11-13 X 31⁄—41⁄;u, borne gan; b, spore-bearing or- on a few-celled, spore-bearing organ 8% $ spores. x 640. about 20-30 X 4—би, slightly curved at first. Fruetifieations more than 6 em. long, more than 115 em. wide. On bark, Maravals, Trinidad, West Indies. This species is characterized by its very thin, velvety fruc- tifieation, pallid smoke-gray with a slight lilae tint, fine hyphae, and small probasidia, spore-bearing organs, and spores. The spore-bearing organs were slightly curved in all cases where spores were attached to them; in the sec- tions some of these organs appeared strongly eurved or hook- shaped but my preparations did not demonstrate this point positively, for the numerous curved hyphal branches were confusing. Specimens examined: Trinidad: Maravals, H. Thaxter, comm. by W. G. Farlow, 28, type. (To be continued.) CATALOGUE OF THE PLANTS OF JASPER COUNTY, MISSOURI (FERNwonrS AND F'LOWERING PLANTS) ERNEST J. PALMER Collector for the Missouri Botanical Garden and the Arnold Arboretum of Harvard University INTRODUCTION This catalogue is based upon collections of plants made by the writer during the years from 1901 to 1913. The serial numbers following each species and variety listed re- fer to the plants collected by me, unless otherwise indicated ; and the specimens are now deposited in the herbarium of the Missouri Botanieal Garden. Many duplieates have been distributed to herbaria and collectors in various parts of this country and abroad. The nomenclature used is, with a few exceptions, that of the seventh edition of Gray’s ‘Manual.’ I believe that this is the first attempt at enumerating the native plants and those growing spontaneously in this part of Missouri. Local catalogues and floras have been published covering several other sections of the state—Jackson County and the vicinities of Louisiana, St. Louis, and Columbia. Im 1886 Professor S. M. Tracy published a list of the plants then known to occur in the state, and in 1895 a list of the woody plants, prepared by Mr. B. F. Bush, was published by the State Horticultural Society. In these two works reference is made to plants of Jasper County. Two of the species mentioned by Professor Tracy as occurring in the county have not been found by the writer. These are Castanea pumila, said to have been collected by Professor Broadhead, and Callirhoe involucrata, which is recorded as having been collected by Letterman. The latter is probably a geographical error. In Mr. Bush’s list 53 woody species are mentioned for Jasper County. All of these, with the exception of Castanea pumila and a few which have been relegated by changes in nomenclature and more recent study, have been confirmed. ANN. Mo. Bor. GARD, Vou. 3, 1916 (345) [VoL. 3 346 ANNALS OF THE MISSOURI BOTANICAL GARDEN But little botanical work had been done in the area prior to the beginning of the writer’s collection. In connection with the work of the State Geological Survey some collections were made by Professor Garland C. Broadhead, who visited Jasper County. Mr. B. F. Bush had also done some collecting about Joplin and Carthage, and possibly others may have collected in the county; but it may be said to have been practically an unknown field, and that it has proved an interesting one is attested by the fact that since the beginning of the present century a number of eminent botanists have visited it. Among these is Dr. C. S. Sargent, the dis- tinguished director of Arnold Arboretum and author of the ‘Silva of North America,’ who has made several trips through the county and has deseribed about 40 new species of trees and shrubs, based wholly or in part upon Jasper County material. Most of these are of the genus Crataegus, the red haws, of which many interesting forms are found here. Mr. Alfred Rehder, of Arnold Arboretum, has also described a new variety of crabapple, Malus %0еп818, var. Palmeri, and an elderberry, Sambucus canadensis, var. sub- mollis, found in our area. Алеха latifolia is a new species of the meadow-beauty collected in Jasper County and de- seribed by Mr. B. F. Bush. Mr. K. K. Mackenzie has re- cently published a description of Geocarpon minimum, a new and anomalous genus, at present known only to occur in our county. Several other plants are still subjects of study and may prove worthy of distinction. In addition to the new species brought to light, a number of previously described plants, but not before collected or known in Missouri, and several that are not given as oc- curring in the range of Gray's ‘Manual’ have been found. Among these may be mentioned Amphilophis Torreyanus, Setaria imberbis, Stenophyllus ciliatifolius, Scirpus carima- tus, Carex arkansana, Crataegus spathulata, var. flavantha, Lathyrus pusillus, Vitis Linsecomii, Myriophyllum proserpina- coides, Daucus pusillus, Lamium purpureum, Hedeoma acin- oides, Diapedium brachiatum, Chrysopsis pilosa, Erigeron tenuis, E. nudiflorus and Marshallia caespitosa. Plants of 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 347 peculiar interest have been submitted to specialists on various groups, and local collections are cited in a number of recently published books and pamphlets, so that Jasper County can no longer be regarded as terra incognita in the botanical world. It is not claimed, however, that the present catalogue is by any means complete or, indeed, anything but a foreword on the flora of the region. While every effort has been made to secure accuracy and to exclude doubtful and invalid species, it is scarcely to be hoped that no errors have crept in. In the present state of botanical science it is highly prob- able that changes in nomenclature and in the taxonomic inter- pretation of certain groups will take place for some years to come; and, no doubt, future investigators will take different views as to the validity of some species here listed. It is very likely, also, that there are a number of plants growing in the county that have so far escaped the attention of the collector. For obvious reasons the immediate vicinity of the writer’s home, in the southwestern part of the county, has been most thoroughly explored. Excursions have been made as opportunity offered into other sections, and considerable collecting has been done in Jasper, Preston, and Sarcoxie Townships, but some portions of the county have received very little attention, especially the northeastern part, which is least accessible at present. The flora of a region, moreover, is not a fixed quantity. Various causes, at present chiefly cultural, are responsible for the introduction of plants from other sections of the country and from foreign lands. On the other hand, as the land comes more and more under cultivation many of the rarer and less hardy species are likely to be exterminated, and some, no doubt, have already disappeared since the settlement of the country by Europeans. The struggle for existence is perhaps as keen in the plant world as in any department of nature. Of the countless number of seeds and spores produced by some species, but a very small per cent find an opportunity to germinate, and a large proportion of the young seedlings are crowded or [Vor. 3 348 ANNALS OF THE MISSOURI BOTANICAL GARDEN starved out, fall a prey to enemies of the animal or plant world, to adverse weather conditions, or to other causes. Those that survive do so, often, by waging a fierce and suecessful battle with other species for the occupation of available ground. In this struggle, which has been going on for countless ages under changing conditions, forms have constantly been modified and those that could not adapt themselves to new environments have been exterminated and replaced by hardier and more fit races. Deep buried in the shales of our coal measures are impres- sions of plants (principally ferns and other cryptogams) that flourished in the tropical marshes of a far-off period. Since then countless races of plants have come and gone, many of them leaving no trace, but under rare, favorable conditions fragments of some of them have been preserved; so that the paleobotanist, digging in the clays and shales of past geological ages, catches here and there a glimpse of those vanished floras, just as the archaeologist delves in the ashes of a ruined city or in the depths of some grass- grown mound in an effort to reconstruct at least an outline of the history of the past. The layman, while recognizing the utility of forestry or the collection and study of plants of cultural, medicinal, or other economie value, is often puzzled to understand the object of purely scientific botany. To the scientist, however, regarding each plant as a wonderful living organism, marvel- ously adapted to its environment, with specific functions to perform, definite relationships with other species, and a history extending back into the remote past, even the com- monest weed is an object of interest and worthy of study. And it is only by their systematic and critical study that a comprehensive knowledge of Nature's methods and the laws of life may be gained. It is important, therefore, that the plants of all parts of the world should be collected, studied, and preserved; and I trust that the following catalogue and the collections upon which it 1s based, although dealing only with the plants 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 349 of a limited area, will be of some value and interest to future investigators. I wish to express my sincere thanks to all of the botanists and friends who have aided and encouraged me in the col- lection and study of the plants of southwest Missouri. To Mr. B. F. Bush, whose unflagging interest in the work from its beginning has been one of the chief stimulants to its prosecution, I am under many obligations, both for the de- termination of plants during successive years when my time and botanical knowledge were both very limited, and for the contribution of literature and specimens for comparison. As the pleasant companion, too, of many botanical excursions I have had the benefit of his unequaled knowledge of plants in the field. I am also greatly indebted to Dr. C. 8. Sargent for the interest he has taken in our woody flora and its in- vestigation. Recognized as the highest authority on North American trees, his publication of several new species from Jasper County has made the region of considerable interest to the botanical world. Thanks are due to Dr. Ezra Brainerd for examining and revising the violets; to Mr. F. W. Pennell for checking the species of Agalinis and allied genera and for his key to this interesting group; to Mr. W. H. Blanchard for valuable notes on the Rubi, and to others. I also wish to make acknowledgments to Dr. George T. Moore, Director of the Missouri Botanical Garden, for affording the oppor- tunity to publish this paper, and to Dr. J. M. Greenman, Curator of the Herbarium, for valuable suggestions and in- terest shown in the work. DESCRIPTION OF JASPER COUNTY Jasper County, Missouri, is situated near the southwestern corner of the state, being in the western tier of counties bor- dering on Kansas, and but the third north from the Arkansas line. Its northern boundary is formed by Barton and Dade Counties, the eastern by Dade and Lawrence, and the south- ern by Newton County. In outline it is nearly rectangular. The width from north to south is about 21 miles, and length [Vor. 8 350 ANNALS OF THE MISSOURI BOTANICAL GARDEN from east to west about 32 miles, the area being 632 square miles. The center of the county is approximately in latitude 37°10’ north, and longitude 94°20’ west of Greenwich. The elevation ranges from about 825 feet in the valley of Spring River near the western boundary to about 1175 feet on the highest hills in the southeastern part. Topographieally, the area is a dissected plain, with hills of low elevation, situated on the western slope of the Ozark dome. Most of the country may be described as an upland prairie with considerable broken, hilly ground bordering the streams, especially along Center Creek in the southern por- tion. Through this plateau the larger streams have carved valleys of varying widths, in the alluvial plains of which they meander from bluff to bluff. These valleys, originally heavily wooded with deciduous forests of oak, maple, ash, walnut and many other trees, are now nearly all cleared and under agriculture. Remnants of the low forests remain only here and there, but considerable areas of the rocky, broken uplands in the southern part are still covered with a virgin or second growth of somewhat stunted timber, in which oaks and hickories predominate. The drainage system is simple, consisting of Spring River and its tributaries, flowing in the main from east to west. The most important of these are North Fork and Dry Fork on the north, and, to the south, Center Creek with its afflu- ents—Jenkins Creek, Jones Creek, and Grove Creek—and Turkey Creek, which flows directly into Spring River a little beyond the Kansas boundary. Spring River and the creeks to the south are perennial streams fed by many springs. North Fork and Dry Fork are intermittent but flow through- out the greater portion of the year. There are no large lakes or other natural bodies of water, but a few bayous or old channels along Spring River and Center Creek afford a habi- tat for some aquatic plants. With the exception of the alluvial valleys the soil is largely residual, resulting from the decomposition of the underlying rocks. In consequence, there is a rather close correlation be- tween the geology and local plant distribution. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 351 The geological formations represented аге the Mississip- pian series, or Subcarboniferous, and the Pennsylvanian se- ries, or Coal Measures. The Mississippian occupies much the greater portion of the area. The rocks of this series con- sist mainly of heavily bedded, semi-crystalline limestones interbedded with lenses of chert, which in places predomi- nates over the limestone. As the silicious rocks are much less soluble than the ealeareous beds, large deposits of angu- lar chert fragments, more or less imbedded in red iron- stained clay, occur locally where the higher beds of lime- stone have been removed by erosion. The horizontal strata of limestone form bold escarpments and bluffs along the river valleys and sometimes outerop through the mantle of residuum on slopes and high hills. The Pennsylvanian formations, consisting of shales and sandstones of the Cherokee group, occupy the northwestern corner of the county, covering portions of Jasper and Twin Groves and the greater part of Preston and Duval Town- ships. Small isolated areas to the southward are too lim- ited in extent to have much influence upon the flora. The rocks of this series, being soft and friable, give rise to a dis- tinct topography with gentle slopes and low hills, through which flow sluggish intermittent streams. The influence upon the flora of these differences in soil and physical features is quite marked. An interesting geological feature is the occurrence of water-worn river gravels covering some of the higher eleva- tions through the south-central portion of the county. These deposits, which have been referred to the Lafayette Gravel of Tertiary age, are of very limited extent, but will be men- tioned in connection with the flora. In so limited an area climatic influences are, of course, essentially uniform, and the variations in altitude are not sufficiently great to affect plant life except indirectly. Over the bulk of the area, underlaid by Mississippian rocks, the factors which determine plant associations and restrict the range of certain species are moisture, shade, and the local : [Vor. 3 352 ANNALS OF THE MISSOURI BOTANICAL GARDEN character of soil and surface rock. The timbered portion may be divided into low woods, bluffs, upland woods, and copses or thickets. The low woods are confined to the alluvial valleys of the larger streams. The lower parts are subject to overflow and farther back from the streams a second terrace or bottom is often found. Characteristic species of the low woods are Carya illinoensis, Salix longifolia, S. nigra, Betula nigra, Quercus macrocarpa, Benzoin melissaefolium, Platanus occi- dentalis, Acer saccharinum, Adelia acuminata, Aristolochia tomentosa, Onoclea sensibilis, Cinna arundinacea, Carex trib- uloides, C. crus-corvi, C. Grayii, var. hispidula, Commelina hirtella, Saururus cernuus, Laportea canadensis, Pilea pu- mila, Polygonum virginianum, Iresine paniculata, Iodanthus pinnatifidus, Arabis dentata, Ranunculus septentrionalis, Ly- copus rubellus, Diapedium brachiatun, Galium V aillantii, Her- acleum lanatum, Sicyos angulatus, Lobelia cardinalis, Eupato- rium coelestinum, and Aster Tradescanti. Some of these spe- cies extend their range through the second bottoms to the bases of the bluffs or into low prairies where moisture is abundant, and mingled with them are many plants more common in the latter situations. The alluvial valleys are usually bounded by cliffs, rarely more than 50 or 60 feet in height, along the slopes and bases of which is found a very characteristic and varied flora, espe- cially where the face of the cliff has a north or east expo- sure. These bluffs are usually well wooded, affording shade and protection for herbaceous species; they are supplied with abundant seepage water, have accumulations of soil washed from the hills above and are enriched by the leaf mould and vegetable humus of ages. On account of their inaccessi- bility and little value for utilitarian purposes they have re- mained more nearly in a primitive state than any other portion of the area. Here are found many plants common to the northeastern states, most of the ferns and orchids, and some of our most beautiful and delicate wild flowers. The list of plants peculiar to this zone is a long one but the fol- 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 353 lowing may be mentioned as typical: Ulmus fulva, Acer sac- charum, Ostrya virginiana, Asimina triloba, Staphylea tri- folia, Adiantum pedatum, Camptosorus rhizophyllus, Poly- stichum acrostichoides, Cystopteris bulbifera, C. fragilis, Botrychium virginianum, Brachyelytrum erectum, Sphen- opholis pallens, Poa Wolfii, Carex sparganioides, C. laxiflora, var. blanda, C. oligocarpa, Arisaema triphyllum, Tradescan- tia virginiana, Erythronium americanum, E. albidum, Smi- lacina racemosa, S. stellata, Polygonatum commutatum, Tril- lium sessile, Cypripedium parviflorum, Corallorrhiza odon- torhiza, Liparis liliifolia, Asarum canadense, Ranunculus re- curvatus, Aquilegia coccinea, Cimicifuga racemosa, Caulo- phyllum thalictroides, Sanguinea Dilleniana, Dicentra Cu- cullaria, Corydalis flavula, Arabis laevigata, Sedum Aer, Euphorbia heterophylla, Erigenia bulbosa, Osmorhiza Clay- toni, O. longistylis, Campanula americana, Solidago arguta, Polymnia uvedalia, and Senecio obovatus, var. rotundus. Where the bluffs have a western or southern exposure and are less shaded a somewhat different type of vegetation ap- pears. Here are found Sapindus Drummondi, Cheilanthes Feei, Notholaena dealbata, Asplenium parvulum, Festuca nutans, Bromus purgans, Uniola latifolia, Camassia escu- lenta, Commelina crispa, Arabis hirsuta, Heuchera hirsuti- caulis, Hypericum cistifolium, Scutellaria cordifolia, Onos- modium hispidissimum, and Lonicera dioica. At the extreme edge of the bluff above may be found Ame- lanchier canadensis, Celtis georgiana, Crataegus obscura, Rhus trilobata, and many of the herbaceous species common to the limestone barrens and dry woods. Above the escarpment a belt is usually occupied by the dry, rocky woods, more or less broken by ravines, up which the rich wood species extend. Characteristic species here are Carya alba, C. megacarpa, C. ovalis and its variety ob- cordata, Quercus stellata, Q. velutina and variety missouri- ensis, Q. marilandica, Cornus florida, Fraxinus americana, Ceanothus americanus, Panicum linearifolium, P. huachucae and the variety silvicola, Danthonia spicata, Gymnopogon 23 [Vor. 8 354 ANNALS OF THE MISSOURI BOTANICAL GARDEN ambiguus, Cyperus ovularis, Carex retroflema, C. varia with variety colorata, Luzula campestris, var. bulbosa, Spiranthes gracilis, Апусма polygonoides, Lespedeza virginica, L. Stu- vei, L. hirta, Desmodium rotundifolium, D. bracteosum, Ascy- rum hypericoides, Lechea villosa, L. tenuifolia, Viola pedata, var. lineariloba, Asclepias quadrifolia, Tephrosia virgimana, Monarda Bradburiana, Scutellaria incana, Aureolaria grandi- flora, Agalinis tenuifolia, Hieracium Gronovii, H. scabrum, Solidago nemoralis, S. radula, S. petiolaris, S. Матай, Aster patens, A. anomalus, A. azureus, A. turbinellus, A. linarii- folius, Erigeron pulchellus, and Parthenium integrifolium. The most extensive and notable floral division is perhaps that of the upland prairies, and here the plants of the plains are the most conspicuous feature. Among prairie plants may be mentioned Andropogon scoparius, A. furcatus, Sor- ghastrum avenaceum, Sporobolus asper, Cyperus acuminatus, Erythromum mesochoreum, Silene regia, Ranunculus fascicu- laris, Draba brachycarpa, Baptisia bracteata, B. australis, I TP purpureum, Astragalus distortus, A. mexi- canus, Desmodium illinoense, D. canadense, Lespedeza cap- itata and variety sericea, Strophostyles pauciflora, Linum me- dium, Euphorbia corollata, Oenothera muricata, O. biennis, Gaura Pitcheri, Polytaenia Nuttallii, Dodecatheon Meadia, Apocynum cannabinum, A. pubescens, Sabatia campestris, As- clepias tuberosa, A. verticillata, A. stenophylla, Acerates flori- dana, A. viridiflora, Phlox pilosa, Lithospermum angustifoli- um, Salvia Pitcheri, Agalinis fasciculata, A. tenuifolia, Hous- tonia minima, Specularia biflora, Prenanthes aspera, Hiera- cium longipilum, Vernonia crinita, Eupatorium altissimum, Liatris scariosa, L. pycnostachya, Aster salicifolius, Silphium lacimatum, S. integrifolium, Parthenium repens, Brauneria pallida, Helianthus scaberrimus, H. grosseserratus, H. mollis, Coreopsis grandiflora, C. palmata, Helenium autumnale, Ca- calia tuberosa, and Cirsium altissimum. Copses and thickets occupy swales or slopes bordering the dry woods, on the one hand, and the upland prairie, on the other. While containing many of the plants of the surround- 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 399 ing regions Ше thickets have quite a distinctive flora, for here are found many shrubs and vines and some herbaceous plants that seldom or never grow in other situations. Among the small trees are many species of Crataegus and Prunus, and a long list of sedges and grasses occur. Ulmus ameri- cana is frequently the only large tree along upland branches, and elsewhere large specimens of Populus deltoides occur far out on flat prairies. Along the branches and in wet de- pressions usually grow Salix Wardi, Amorpha fruticosa, Cornus Amomum, Penthorum sedoides, Hibiscus incanus, H. militaris, Cicuta maculata, Asclepias incarnata, Lobelia siphil- itica, and Lycopus americanus. Occupying somewhat drier situations are Celtis occidentalis, Corylus americana, C. ros- trata, Ribes missouriensis, Opulaster intermedius, Malus toensis, var. Palmeri, Rubus occidentalis, В. Andrewsianus, R. canadensis, Rosa setigera, Prunus hortulana, P. Munsoni- ana, Cercis canadensis, Zanthoxylum americanum, Ptelea trifoliata, Rhus glabra, R. copallina, Evonymus atropurpur- eus, Rhamnus lanceolata, Cornus asperifolia, C. Baileyi, Vi- burnum prunifolium, V. rufidulum, Symphoricarpos orbicu- latus, Smilax rotundifolia, S. hispida, Clematis Pitcheri, Menispermum. canadense, Cocculus carolinus, Celastrus scan- dens, Vitis cinerea, Ampelopsis cordata, Geum vernum, G. canadense, Cassia Medsgeri, Phaseolus polystachyus, Stro- phostyles umbellata, Galactia volubilis, Vincetoxicum caro- linense, V. Baldwinianum, Scrophularia marilandica, Dasis- toma macrophylla, Galium pilosum, G. circaezans, Triosteum perfoliatum, Rudbeckia triloba, Verbesina virginica, and Ca- calia atriplicifolia. In addition to the area, of which the main floral zones are outlined above, there are several rather distinct regions in which a close association between the underlying geological features and the present plant life is clearly traceable. The most extensive of these is the sand hill region or Barton upland, underlaid by Pennsylvanian strata, in the northwestern part of the county. This section is largely high prairie which slopes down to, and is limited by, the valleys [Vor. 3 356 ANNALS OF THE MISSOURI BOTANICAL GARDEN of North Fork on the east and of Spring River on the south. Seepage springs and small, sluggish streams give a wet and somewhat swampy character to some portions. The soil is, for the most part, a sandy loam, with large blocks of sand- stone on the surface of some of the higher hills and sand- stone and shale outeropping along streams. Peculiar to this area, so far as has been observed within the county, are Andropogon ternarius, Panicum scoparium, Setaria imberbis, Festuca Shortti, Eleocharis ovata, Rynchospora cymosa, Scleria ciliata, Scirpus carinatus, Carex arkansana, Juncus monostichus, J. effusus, J. polycephalus, J. robustus, Poly- допит sagittatum, Froelichia gracilis, Geocarpon minimum, Ranunculus oblongifolius, Rhexia latifolia, Proserpinaca palustris, Centunculus minimus, Chrysopsis pilosa, and Cir- sium discolor. Other species, more or less characteristic of this region but which have also been found elsewhere in the county, are Quercus palustris, Cyperus esculentus, Fimbris- tylis castanea, var. puberula, Rynchospora glomerata, Scleria triglomerata, Carex umbellata, C. stipata, Luzula campestris, var. bulbosa, Polygonum tenue, Anemone caroliniana, Poly- gala sanguinea, P. incarnata, Crotonopsis linearis, Viola pedata, var. lineariloba, V. sagittata, Monarda mollis, Linaria canadensis, Castilleja coccinea, and Marshallia caespitosa. Of the above list, Froelichia gracilis and Chrysopsis pilosa have only been found on an isolated sandstone hill at the state line, near Smithfield, and Scirpus carinatus in lower ground near the same place. A region of much more limited extent, but with a distinet flora, is situated in the valley of Turkey Creek, two miles B EN of Joplin, where the Grand Falls chert of the Mississippian series outcrops. The surface of the chert is irregular, with hummocks and basin-like depressions of va- rious sizes. Many of the latter contain thin layers of soil washed from the higher ground and in wet times are filled with rain-water, which, since the rock where unfractured is impermeable, is retained until evaporated by the sun. The superabundanee of moisture in rainy seasons and extreme Ф 19161 PALMER—PLANTS ОЕ JASPER COUNTY, MISSOURI 857 dryness at other times, together with an almost total absence oÍ shade, the sparsity of soil and other ecological factors, produces peculiar conditions clearly reflected in the flora, which is partly hydrophytie but mainly xerophytie and suc- culent. Some of the characteristic species are Cheilanthes lanosa, Selaginella rupestris, Leptochloa fascicularis, Cy- perus aristatus, Digitaria filiformis, Aristida basiramea, Al- lium mutabile, Polygonum tenue, Talinum parviflorum, T. calycinum, Portulaca pilosa, Selenia aurea, Sedum Nuttal- lianum, S. pulchellum, Lathyrus pusillus, Crotonopsis line- aris, Opuntia macrorhiza, Spermolepis echinata, Linaria canadensis, Specularia leptocarpa, Coreopsis tinctoria, and Krigia occidentalis. The chert formation is much more ex- tensively exposed a few miles to the south along Shoal Creek in Newton County, where all of the above and several other peculiar species are found. Somewhat similar, though quite distinct, are the limestone barrens found at several places in the county, where horizon- tal strata of the Mississippian series appear on the surface. All of these are of quite limited extent but support a charac- teristic flora. Some of the best-marked localities are the fol- lowing: one mile north of Jasper, on south side of Coon Creek; two miles southwest of Neck City, along a small branch; one mile north of Carterville, on south side of Center Creek and along a branch south and east of Carterville and Webb City. Typical plants of these limestone barrens are Ophioglossum Engelmann, Sporobolus pilosus, Sphenopholis obtusata, Bouteloua curtipendula, Cyperus aristatus, Camas- sia esculenta, Oxybaphus albidus, Arenaria patula, Talinum calycimum, Portulaca pilosa, Draba cuneifolia, Corydalis montana, Sedum pulchellum, Acalypha gracilens, Tragia ramosa, Euphorbia missouriensis, Malvastrum angustum, Mentzelia oligosperma, Opuntia humifusa, Chaerophyllum texanum, Heliotropium tenellum, Isanthus brachiatus, Galium virgatum, Aster oblongifolius, var. rigidulus, and Artemisia mewicana. e Palmer, E. J. Flora of the Grand Falls chert barrens. Acad. Sci. St. Es Trans. 19:97-112. 1910. [Vor. 3 358 ANNALS OF THE MISSOURI BOTANICAL GARDEN Some high hills near Prosperity and Duenweg, partly cov- ered with deposits of Lafayette gravel, have a somewhat charaeteristie flora, although few, if any, of the species are peculiar to them. Antennarias are a conspicuous feature of the spring vegetation, and it is probable that one or more undescribed species occur here. Other plants that may be mentioned are Panicum depauperatum, P. Werneri, Carex umbellata, C. Meadii, Fimbristylis castanea, var. puberula, Stipa spartea, Viola pedata, var. lineariloba of which a white form is frequent, V. sagittata, Lithospermum canescens, Sene- cio plattensis, and Marshallia caespitosa. Introduced plants, while quite numerous, do not as yet form a large percentage of the flora, but their invasion is steadily increasing, especially in the western part of the county where many railroads enter. Grasses and common weeds form the bulk of these emigrants. These flourish largely in waste places and in cultivated ground, and their distribution has little definite relationship to the native plants. Among species of recent introduction a few show a tendency to become wide-spread. Lespedeza striata is be- coming common in dry rocky woods, and the white-flowered sweet clover, Melilotus alba, is frequently found along road- sides and railroads. The sand burr, Solanum rostratum, has in recent years become quite common in ballast or waste ground, but it shows little tendency to spread beyond such situations. Helenium tenuifolium is beginning to appear along railroads and bids fair to become a nuisance. Perilla frutescens is established at several stations and, judging by the rapidity with which it has spread in the bottoms of many Ozark streams, is likely to become common. Perhaps most pernicious of all is the Johnson grass, Sorghum halepense, which is established in a number of low fields. From a study of the above and the following list it will be seen that the flora of Jasper County is a diverse and com- posite one. The plants of the near plains are perhaps the dominant element and the most striking feature, but mingled with them are a number of types of the northeastern states, 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 309 others characteristic of the Ozark or southeastern mountain region, quite an element of southwestern species and some that are distinctly southern. That the central Mississippi Valley is the meeting ground of these various floras is well known, and Jasper County, through its diversity of soil and topography, fairly epitomizes, and is typical of, the region. CATALOGUE OF SPECIES POLYPODIACEAE NOTHOLAENA DEALBATA (Pursh) Kunze. 640, 641, 642, 2992. ADIANTUM PEDATUM L. 634, 635, 686, 1319. PTERIS AQUILINA, var. PSEUDOCAUDATA Clute. 643, 644, 645. CHEILANTHES LANOSA (Michx.) Watt. 1321, 2376. CHEILANTHES rer Moore. 649, 650, 651, 1827, 2991. PELLAEA ATROPURPUREA (L.) Link. 637, 638, 639, 2985, 2989, 3085. ASPLENIUM PARVULUM Mart. & Gal. 628, 629, 2993, 3168. ASPLENIUM PLATYNEURON (L.) Oakes. 631, 632, 758, 2481, 2019, 8152. ASPLENIUM PLATYNEURON, var. SERRATUM (Miller) BSP. 2085. CAMPTOSORUS RHIZOPHYLLUS (L.) Link. 625, 626, 1543. POLYSTICHUM ACROSTICHOIDES ( Michx.) Schott. 622, 623, 2961. CvsroPTERIS BULBIFERA (L.) Bernh. 647, 2964, 2977, 3169. This species is sometimes difficult to distinguish from Cys- topteris fragilis, as it seldom produces the characteristic fleshy bulblets in our area, and the fronds seldom exceed 20 em. in length, but it differs from the next species in being broader at the base, thinner, and with segments less decur- rent along the rachis. It is found along moist shaded ledges of limestone bluffs. C. fragilis seems to prefer rich banks and shaded hillsides. Сувтортентв FRAGILIS (L.) Bernh. 646, 2327, 2978, 3018, 3255, Woopsia oBTUSA (Spreng.) Torr. 619, 620, 2216, 2328, 2480, 3059. ONOCLEA sENSIBILIS L. 7320, 2111, 2976, 3156, 3697. [Vor. 3 360 ANNALS OF THE MISSOURI BOTANICAL GARDEN OPHIOGLOSSACEAE OPHIOGLOSSUM ENGELMANNI Prantl. 2044, 2942, 3358. Engelmann's adder’s-tongue has been found in thin soil of limestone barrens near Carterville and in similar situ- ations along North Fork of Spring River near Neck City. At the latter station several large colonies were growing. Вотвуснитм VIRGINIANUM (L.) Sw. 616, 617, 618, 2030, 2947, 3154, 3698, 3359. BornvcuruM oBLIQUUM Muhl. 3153. Rare and local. A few plants were found in rich hillside woods at a station along Center Creek, about four miles southeast of Carthage, September 11, 1910. EQUISETACEAE EQUISETUM ARVENSE L., 890, 2544. FQUISETUM HYEMALE L. 958. SELAGINELLACEAE SELAGINELLA RUPESTRIS (L.) Spring. 1322. PINACEAE JUNIPERUS VIRGINIANA L. 887, 1919. The red cedar is rare in our county. A number of small trees were noted on rocky uplands north of Spring River between Carthage and Alba; some large stumps were found along a bluff of North Fork between Jasper and Preston, but no living plants were left at this station. Young plants are occasionally found in woods throughout, the seeds prob- ably having been carried by birds from planted trees. TYPHACEAE 'TYPHA LATIFOLIA L. 2306. SPARGANIACEAE SPARGANUM AMERICANUM Nutt. 780, 3158, 3159. POTAMOGETON OBTUSIFOLIUS Mertens & Koch. 3184. PorAMoGETON FoLIosus Raf. 3251. POTAMOGETON DIMORPHUS Raf. 1257, 2514, 3879. POTAMOGETON PECTINATUS L. 2204, 3251. ALISMACEAE SAGITTARIA LATIFOLIA Willd. 460, 809, 1080. 1916] PALMER—PLANTS 0F JASPER COUNTY, MISSOURI 361 SAGITTARIA AMBIGUA J. G. Sm. 2130, 3061, 9727. SAGITTARIA GRAMINEA Michx. 3677. LoPHOoTOCARBPUS CALYCINUS (Engelm.) J. С. Sm. 471, 3748. ALISMA PLANTAGO-AQUATICA L. 343, 2183, 2783, 3786. HYDROCHARITACEAE EropEA CANADENSIS Michx. 3765. GRAMINEAE TRIPSACUM DACTYLOIDES L. 1372, 3411, 3762. ANDROPOGON SCOPARIUS Michx. 220. ANDROPOGON VIRGINICUS L. 1007. ANDROPOGON Ешллоттп Chapman. 2808. ANDROPOGON TERNARIUS Michx. 3268. ANDROPOGON FURCATUS Muhl. 678, 2591, 2838, 3080, 3143, 3183. ANDROPOGON CHRYSOCOMUS Nash. 217, 678. AMPHILOPHIS TorrEyanus (Steud.) Nash. SoRGHASTRUM NUTANS (L.) Nash. 279. SORGHUM HALEPENSE (L.) Pers. 997. SORGHUM VULGARE L. 637. DIGITARIA FILIFORMIS (L.) Koeler. 2810, 3116. DIGITARIA HUMIFUSA Pers. 2815. DIGITARIA SANGUINALIS (L.) Scop. 607, 3823. Leprotoma COGNATUM (Schultes) Chase. 978, 1403, 3112. PASPALUM MUCRONATUM Muhl. 1389, 1490. PASPALUM DISSECTUM L. 968, 969, 1393, 3227, 3878. PASPALUM MUHLENBERGII Nash. 1117, 1390, 2341, 3066, 3161, 2289. PASPALUM LAEVE Michx. 1118. PASPALUM ANGUSTIFOLIUM LeConte. 777, 3067. PASPALUM PRAELONGUM Nash. 966, 1439, 3140, 3228. PASPALUM FLORIDANUM Michx. 1391, 1392, 3170. PASPALUM LAEVIGLUME Scribn, 967, 2334. PASPALUM GLABRATUM (Engelm.) Mohr. 965, 970. PANICUM CAPILLARE L. 227, 1402, 2550, 3090, 3096. PANICUM GarrTINGERI Nash. 3204. PANICUM FLEXILE (Gattinger) Scribn. 786, 838, 843, 1119, 1400, 3089. Ë [Vor. 3 362 ANNALS OF THE MISSOURI BOTANICAL GARDEN PANICUM PHILADELPHICUM Bernh. 815, 3115, 2447. PANICUM DICHOTOMIFLORUM Michx. 2647, 3119, 3225. PANICUM viRGATUM L. 216, 975, 977, 992, 2439, 2533. PANICUM AGROSTOIDES Spreng. 972, 1367, 3025, 3174. PANICUM ANCEPS Miehx. 606,764, 1406, 3048. PANICUM DEPAUPERATUM Muhl. 847, 1881, 1953. PANICUM PERLONGUM Nash. 594, 2156, 3372. PANICUM LINEARIFOLIUM Seribn. 2236, 3382, 3354 (1555 of В. Е. Bush). PANICUM WERNERI Scribn. 2426. Panicum HUACHUCAE Ashe. 749, 846, 2047, 2101, 3753. PANICUM HUACHUCAE, Var. siLVICOLA Hitehe. & Chase. 1394, 1399, 2017, 3732, 3746, 3756. PANICUM SUBVILLOSUM Ashe. 3398, 3402. PANICUM TENNESSEENSE Ashe. 596, 974, 3409, 3761, 3714. PANICUM PnAECOCIUS lHitehe, & Chase. 2144. PANICUM SPHAEROCARPON ЕП. 247, 850, 1398, 2154, 2507, 3808. PANICUM SCRIBNERIANUM Nash. 223, 590, 2046, 2014, 2160, 2157, 9397, 3713. Panicum scoPARIUM Lam. 611, 1401, 2293. PANICUM CLANDESTINUM П. 971, 1404, 1395. Panicum PuBIFOLIUM Nash. 1562. PANICUM LATIFOLIUM L. 597, 849, 1405, 740, 2106, 2966. Panicum Непьевг Nash. 973, 1857. Panicum LINDHEIMERI Nash. 748, 1396, 2179, 2276, 3037. ECHINOCHLOA CRUSGALLI (L.) Beauv. 246, 976, 2570, 3486, 3477. SETARIA IMBERBIS R. & S. 2438, 2471, 3050, 3422, 3034. SETARIA GLAUCA (L.) Beauv. 2347, 3071. ÑETARIA viRIDIS (L.) Beauv. 606, 3076. SETARIA ITALICA (L.) Beauv. 2551. SETARIA ITALICA, var. GERMANICA (Mill.) Richter. 991. CENCHRUS TRIBULOIDES L. 763. LxrnstA VIRGINICA Willd. 1000, 1561, 1381, 3207. LEERSIA овугошев (L.) Sw. 1001, 1370, 1557, 2682. PHALARIS CAROLINIANA Walt. 221. PHALARIS CANARIENSIS L. 3754. БТТРА SPARTEA Trin. 2159. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 363 ARISTIDA DICHOTOMA Michx. 1387, 1494, 3488. ARISTIDA BASIRAMEA Engelm. 2854. ARISTIDA GRACILIS Ell. 3217. ARISTIDA INTERMEDIA Scribn. & Ball. 1121. ARISTIDA OLIGANTHA Michx. 814, 1385, 2527. ARISTIDA PURPURASCENS Poir. 1002, 1386. ARISTIDA FASCICULATA Torr. 4361. MUHLENBERGIA SOBOLIFERA (Muhl.) Trin. 856, 1375, 2529, 2546. MUHLENBERGIA TENUIFLORA ( Willd.) BSP. 218, 2734. MUHLENBERGIA SYLVATICA Torr. 3187. MUHLENBERGIA MEXICANA (L.) Trin. 1388, 2686, 2797, 3463, 3467, 3475. MvHLENBERGIA ScHREBERI J. F. Gmel. 1376, 2646, 2984. MUHLENBERGIA DIFFUSA Schreb. 988, 990, 3488. MUHLENBERGIA CAPILLARIS ( Lam.) Trin. 2828. BRACHYELYTRUM ERECTUM (Schreb.) Beauv. 2459, 2545, 3196. PHLEUM PRATENSE L. 995; 2127. ALOPECURUS GENICULATUS L. 225, 1826. SPOROBOLUS CANOVIRENS Nash. 1005. Sporopouus AsPER ( Michx.) Kunth. 2811, 3117, 3134, 3215, 3252, 3216. SpoRoBOLUS VAGINIFLORUS (Torr.) Wood. 2796, 3118. SporoBoLus NEGLECTUS Nash. 2843. SPOROBOLUS PILOSUS Vasey. 987, 3133. SponoBoLUs Овоммохри ( Trin.) Vasey. 3479, 3485. AGROSTIS ALBA L. 860, 2223. Acrostis ErrroTTIANA Schultes. 1377, 2941. Асвовтів HYEMALIS ( Walt.) BSP. 2288, 2517. AGROSTIS PERENNANS (Walt.) Tuckerm. 835, 854, 979, 993, 2410, 3008, 3416. AGROSTIS INTERMEDIA Scribn. 3101. CINNA ARUNDINACEA L. 857, 998, 1368, 2492. SpHENOPHOLIS oBTUSATA (Michx.) Seribn. 2444, 2987. SpHENOPHOLIS PALLENS (Spreng.) Scribn. 2948, 2962, 3363. KoELERIA CRISTATA (L.) Pers. 222, 2970. DaxTHONIA SPICATA (L.) Beauv. 1364, 2224. Spartina MICHAUXIANA Hitche. 1369, 1495, 2573. [Vor. 3 364 ANNALS OF THE MISSOURI BOTANICAL GARDEN Cynopon Пастугок (L.) Pers, 2450, 3501. SCHEDONNARDUS PANICULATUS (Nutt.) Trel. 2497. GyMNoPocox AMBIGUUS ( Michx.) BSP. 1371, 1373, 2829. BOUTELOUA cuRTIPENDULA (Michx.) Torr. 215, 2340, 2359, 2612. EuLEUSINE INDICA Gaertn. 605. LEPTOCHLOA ATTENUATA Nutt. 2577, 2733, 3094, 3911. LEPTOCHLOA MUCRONATA (Michx.) Kunth. 2569. LEPTOCHLOA FASCICULARIS (Lam.) Gray. 2509. Tripens strictus (Nutt.) Nash. 1374, 2627, 2812, 3065. TRIDENS FLAVUS (L.) Hitehe, 2621, 3195. Енлавовтів nyPNorpEs (Lam.) BSP. 855, 1378, 1379. ERAGROSTIS CAPILLARIS (L.) Nees. 231, 994, 1380, 2267, 2448, 2853, 2836, 3028, 3230. Enacnosris Frangi (Fisch, Mey. & Lall.) Steud. 2818. EnacnosrIS PrLosA (L.) Beauv. 980, 2456, 2489, 3008. Eracrostis Ровѕни Schrad. 980, 3138, 3279. ERacRosTIS MEGASTACHYA (Koeler) Link. 229, 600. ERAGROSTIS PECTINACEA (Michx.) Steud. 842, 996, 1146, 2449, 2852. MELICA mutica Walt. 861, 1809, 3769. Me ica NITENS Nutt. 859. DIARRHENA DIANDRA ( Michx.) Wood. 3803. UNIOLA LATIFOLIA Michx. 226, 603, 1373. ПАСТУ118 GLOMERATA L. 598. Poa annua L. 576, 986. Poa CHapMANIANA Seribn. 554, 2911, 3369. Poa CoMPRESSA L. 2451. Poa PRATENSIS L. 985, 1383. Poa SYLVESTRIS Gray. 987, 1382, 1384, 1836. Poa Wotrn Scribn. 853, 3364. Guycerta NERVATA ( Willd.) Trin. 2068, 2273, 3744, 3780. Frstuca ocTOFLORA Walt. 224, 613, 1882, 3664, 2687. FESTUCA ELATIOR L. 2369, 2408, 2495, 3077. Festuca NUTANS Spreng. 858, 999, 2240, 2322, 2413, 3656, 3693, 3364. Festuca Suorti Kunth. 2246, 2429, 2431, 2432. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 365 Bromus sEcALINus L. 589. Bromus pureans L. 1142, 1365, 1366, 2239. Bromus commutatus Schrad. 3413, 3721. Bromus ARVENSIS L. 2979. AGRoPYRON SMITHII Rydb. 2590. AGROPYRON REPENS (L.) Beauv. 3418, TRITICUM VULGARE L. 2090. Ноновом JUBATUM L. 2222. Нововом PUSILLUM Nutt. 984. Ешумту vircinicus L. 228, 2477, 3024. ELYMUS GLABRIFLORUs (Vasey) Seribn, & Ball. 983, 1143. ELYMUS CANADENSIS L. 588, 2333. ErnvMvs eLAUvCUS Buckley. 3045. ELYMUs sRACHYSTACH YS Scribn. & Ball. 982, 2453, 2534. Hysrrix PATULA Moench. 612, 981, 3781. CYPERACEAE CYPERUS RIVULARIS Kunth. 1047, 2754. CYPERUS ARISTATUS Rottb. 884, 2520, 3088, 3280. CYPERUS ACUMINATUS Torr. & Hook. 242, 2264, 3229. CYPERUS PSEUDOVEGATUS Steud. 234, 1048. CYPERUS ESCULENTUS L. 2482, 3141, 3447, 3476. CYPERUS ESCULENTUS, Var. ANGUSTISPICATUS Boeckl. 2601. CYPERUS sPECIOsUS Vahl. 3224. CYPERUS STRIGOSUS L. 245, 3213, 3796, 3462. CYPERUS STRIGOSUS, Var. ROBUSTIOR Kunth. 241, 3142. CYPERUS STRIGOSUS, var. coMPosrrUS Britton. 2712, 3245. CYPERUS STRIGOSUS, Var. CAPITATUS Boeckl. 1049. CYPERUS Lancastrriensis Porter. 1493, 2476, 2531. CYPERUS OVULARIS ( Michx.) Torr. 232, 2366, 3794. CYPERUS FILICULMIS Vahl. 233, 2265, 3238. Cyperus Воѕни Britton. 2946. KYLLINGA PUMILA Michx. 788, 3834. Exeocuaris ovata (Roth) В, & S. 1362. Ешкоснавів oprusA ( Willd.) Schultes. 690, 1363, 3090, 3678, 3750. ELEOCHARIS ENGELMANNI Steud. 189, 1563, 3631, 3770. [Vor. 3 366 ANNALS OF THE MISSOURI BOTANICAL GARDEN Errocnanis PALUSTRIS (L.) R. & S. 3379, 2168, 2169, 3718. ELEOCHARIS PALUSTRIS, Var. GLAUCESCENS ( Willd.) Gray. 1817. Еткоснавнів rENUIS ( Willd.) Schultes. 765, 2013, 3444. ErrocHanis ACUMINATA ( Muhl.) Nees. 3630. ELEOCHARIS LANCEOLATA Fernald. 2292. ELEOCHARIS МАСКОЅТАСНҮА Britton. 2149, 2564. ErEocHARIS ACICULARIS (L.) В, & S. 2587, 3421, 3426, 3719. STENOPHYLLUS CAPILLARIS (L.) Britton. 3029, 3265, 3278. STENOPHYLLUS CILIATIFOLIUS (L.) Mohr. 776, 1361, 3212. FrMBRISTYLIS CASTANEA, Var. PUBERULA (Michx.) Britton. 595, 742, 1963, 2012, 2123, 2296, 3624. FrwBRISTYLIS LAXA Vahl. 1046, 3120, 3264. FIMBRISTYLIS AUTUMNALIS (L.) В, & Ө. 2360. ScIRPUS AMERICANUS Pers. 274, 677, 2496. This species of rush frequently grows about the mines, in wet ‘‘mineral sand,’’ and in water pumped from mines which is strongly impregnated with iron sulphide and is fatal to many forms of plant life. SCIRPUS vALIDUS Vahl. 608, 2167. SCIRPUS ATROVIRENS Muhl. 2128, 2202. SCIRPUS LINEATUS Michx. 593, 2091. SCIRPUS CARINATUS Gray. 3642. RvNcHosPoRA cymosa Ell. 2440, 2473, 3038, 3051. RYNCHOSPORA GLOMERATA (L.) Vahl. 1044, 1045, 2434, 3038a, 3056. SCLERIA TRIGLOMERATA Michx. 900, 2155, 2474. SCLERIA CILIATA Miehx. 2472, 2471, 3033. CAREX scoPARIA Schkuhr. 2291. CAREX TRIBULOIDES Wahlenb. 864, 2187, 3740, 3772. CAREX MIRABILIS Dewey. 3771. CAREX HORMATHODES Fernald. 3374. CAREX HORMATHODES, var. тхуівА (W. Boott) Fernald. 3662. Carex Віскхклді Britton. 1958, 2284, 3674, 3716, 3724. Carex FESTUCACEA Schkuhr. 555, 869, 874, 2150, 3370, 3582, 3653, 3760. Carex ROSEA Schkuhr. 871. CAREX ROSEA, var. RADIATA Dewey. 3385, 3635, 3692. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 367 CAREX RETROFLEXA Muhl. 876, 1354, 1861, 3353. CAREX AUSTRINA (Small) Mack. 1353, 2225, 3652, 3684, 3751. CAREX ARKANSANA Bailey. 3723. CAREX CEPHALOPHORA Muhl. 880, 3633. Carex Leavenwortui Dewey. 867, 872, 1358, 1778, 1851. CAREX SPARGANIOIDES Muhl. 1356, 3691. CAREX vULPINOIDEA Michx. 1052, 2067, 2126, 3417. САВЕХ CONJUNCTA Boott. 877, 3696, 8787. CAREX ANNECTENS Bicknell. 3759. Carex STIPATA Muhl. 873, 1962, 2174, 2442, 3679. CAREX cRU$S-CORVI Shuttlw. 863, 2197. Carex Емокут Dewey. 1910, 3706. Carex HIRSUTA Willd. 1593, 1865, 2078, 2206, 3720, 3675, 3807. Carex Вовни Mack. 3389, 878, 1865, 3676. CAREX CAROLINIANA Schwein. 2912, 3743. CAREX AGGREGATA Mack. 2207. Carex Davisn Schwein. & Torr. 1352, 1852, 3403, 3699. Carex SHorTIANA Dewey. 870, 2193, 3375, 3670. CAREX UMBELLATA Schkuhr. 238, 3589, 3625. Carex VARIA Muhl. 1357, 1666. CAREX VARIA, var. COLORATA Bailey. 578, 3520, 3602. Carex Мварп Dewey. 239, 553, 1763, 3372, 3627, 3638. САВЕХ PTYCHOCARPA Steud. 2266. CAREX LAxIFLORA Lam. 599, 866, 868. CAREX LAXIFLORA, Var. PATULIFOLIA (Dewey) Carey. 1880. CAREX LAXIFLORA, var, VARIANS Bailey. 3366. CAREX LAXIFLORA, Var. BLANDA (Dewey) Воо. 3694. CAREX OLIGOCARPA Schkuhr. 1953, 3364. Carex GRISEA Wahlenb. 882, 1050, 3700, 3773. CanEX GRISEA, Var. RIGIDA Bailey. 1818. Carex LANUGINOSA Michx. 1874, 3741. Carex RIPARIA W. Curtis. 1820, 1909, 2293. Carex SQUARROSA L. 875, 3403, 3735. Carex Ғвахкп Kunth. 602, 1051, 2192. Carex LURIDA Wahlenb. 2135, 3054. CAREX LUPULINA Muhl. 1050, 3017. CAREX LUPULINA, Var, PEDUNCULATA Dewey. 864, 2191, 2305, 3626, 3768. | [Vor. 3 368 ANNALS OF THE MISSOURI BOTANICAL GARDEN CAREX GRAYII, var. HISPIDULA Gray. 604, 1359, 2023, 2196, 3739. CAREX OKLAHOMENSIS Mack. 3405. This species was deseribed by Kenneth K. Mackenzie! giv- ing reference to No. 3405 of this collection. ARACEAE ARISAEMA TRIPHYLLUM (L.) Schott. 548, 687. AnrsAEMA Dracontium (L.) Schott. 519. Acorus CALAMUS L. 1716, 2505. LEMNACEAE SPIRODELA POLYRHIZA (L.) Schleid. 1247, 2200. COMMELINACEAE TRADESCANTIA BREVICAULIS Raf. 298. I have grown hybrids of this and the following species in my garden. TRADESCANTIA REFLEXA Raf. 299,657, 1954, 3365. A form of this species with sheaths and leaves copiously hir- sute has been found in rocky woods. TRADESCANTIA VIRGINIANA L. 1289. COMMELINA VIRGINICA L. 300, 2380. COMMELINA HIRTELLA Vahl. 1092, 1288, 1485. COMMELINA NUDIFLORA L. 1287, 1486 COMMELINA CRISPA Wooton. 2242. CoMMELINA COMMUNIS L. 2311. PONTEDERIACEAE PowTEDERIA CORDATA L. 1087. HETERANTHERA LIMOSA (Sw.) Willd. 829, 2623. JUNCACEAE Juncus rENvIS Willd. 610, 750. Juncus INTERIOR Wiegand. 591, 2118, 2446. Juncus MowosTICHUS Bartlett. 2285. JUNCUS EFFUSUS L. 2283. JUNCUS POLYCEPHALUS Michx. 2300, 3430. Juncus Noposus L. 609. Juncus Torreyi Coville. 601, 2504. 1Тоггеуа 14:125-127. 1914. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 369 Juncus BRACHYCARPUS Engelm. 237, 2103, 2119, 2151, 2212, 2286, 2299, 2484. Juncus DIFFUsISsIMUS Buckley. 236, 2211, 3757. Juxcvs вововтов (Engelm.) Coville. 2301. Juncus MARGINATUS Rostk. 240, 751, 2275. JUNOUS MARGINATUS, Var. SETOSUS Coville. 243, 2122. JUNCUS ARISTULATUS Michx. 1408, 2117, 2294. LUZULA CAMPESTRIS, var. BULBOSA A. Wood. 577, 1407, 1604, 3644. LILIACEAE MELANTHIUM VIRGINICUM L. 2182. ALLIUM CANADENSE L. 521, 2099. ALLIUM MUTABILE Michx. 2114, 2274. ALLIUM sativum L. 2129, 2503. NorHOSCORDUM BIVALVE (L.) Britton. 294. LILIUM CANADENSE L. 2354, 2891. ERYTHRONIUM AMERICANUM Ker. 292, 1623. ERYTHRONIUM ALBIDUM Nutt. 1613, 1617. EnvTHRONIUM MESOCHOREUM Knerr. 293, 1596, 1605. C'AMASSIA ESCULENTA ( Ker.) Robinson. 291. CONVALLARIACEAE SMILACINA RACEMOSA (L.) Desf. 737. SMILACINA STELLATA (L.) Desf. 556, 736. Рогхаохаттум coMMUTATUM (R. & S.) Dietr. 735. TRILLIUM SESSILE L. 295. ASPARAGUS OFFICINALIS L. 2312. SMILAX HERBACEA L. 473. SMILAX ECIRRHATA (Engelm.) Wats. 487, 736. SMILAX ROTUNDIFOLIA L. 55, 735, 892. Бмпах Bona-nox L. 340, 718. Plants apparently belonging to this species are not uncom- mon, but I have not collected it in fruit in the county. SMILAX HISPIDA Muhl. 1074, 3850. DIOSCOREACEAE DIOSCOREA PANICULATA Michx. 527. 24 [VoL. 3 370 ANNALS OF THE MISSOURI BOTANICAL GARDEN Names and determinations of H. H. Bartlett! are followed for this species and succeeding variety. DIOSCOREA PANICULATA, Var. GLABRIFOLIA Bartlett. 832. AMARYLLIDACEAE Hypoxis uirsuta (L.) Coville. 694, 695. IRIDACEAE Tris vERSICOLOR L. 883, 2112. BELAMCANDA CHINENSIS (L.) DC. 1095. SISYRINCHIUM CAMPESTRE Bicknell. 119, 560, 686. SISYRINCHIUM GRAMINEUM Curtis. 120, 121, 122. ORCHIDACEAE CYPRIPEDIUM PARVIFLORUM Salish. 895, 1526, 1860. PoGoNIA TRIANTHOPHORA (Sw.) BSP. 3102. SPIRANTHES GRACILIS (Bigel.) Beck. 797, 31.22. SPIRANTHES VERNALIS Engelm. & Gray. 408, 2566. SPIRANTHES CERNUA (L.) Richard. 337, 826, 1484, 3225, 3263. CORALLORRHIZA IZA Nutt. 1267, 1914. LIPARIS LILIIFOLIA (L.) Richard. 2171. PIPERACEAE SaAvRURUS CERNUUS L. 793, 1131. SALICACEAE SALIX NIGRA Marsh. 921, 2002. SALIX LONGIPIPES Anders. 917, 919. SALIX ALBA, Var. VITELLINA (L.) Koch. 1246, 2606. SALIX LONGIFOLIA Muhl. 920. SALIX HUMILIS Marsh. 386,918, 1611. SALIX CORDATA, var. MYRICOIDES ( Muhl.) Carey. 1875, 2843. PoPULUS ALBA L. 390. POPULUS DELTOIDES, var. MISSOURIENSIS Henry. 22, 2100, 3321, 3361, 3380, 3504, 3598. PoPutLus NIGRA L. 1878. 1U, 8. Dept. Agr., Bul. 189. 1910. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 371 JUGLANDACEAE JUGLANS NIGRA L. 960, 2071. Carya Pecan C. K. Schneider. 1313. The pecan is not abundant in this section. Trees are found occasionally along Spring River and the lower part of Cen- ter Creek, near Carl Junction and Smithfield. Carya ovata (Mill. K. Koch. 456, 1314, 1904. CARYA ALBA (L.) K. Koch. 65, 1072. CARYA ALBA, Var. FICOIDES Sarg. 3493. This variety of the mocker-nut, with large pear-shaped fruit, was described by Professor Sargent! from a tree growing in Mt. Hope Cemetery, Webb City. It has been noted in other parts of southwest Missouri. Carya ovais ( Wang.) Sarg. 14, 25, 818, 1073, 1127, 2809, 3493. CARYA OVALIS, Var. OBCORDATA Sarg. | CARYA CORDIFORMIS ( Wang.) К. Koch. 26, 1126. CARYA CORDIFORMIS, Var. LATIFOLIA Sarg. CARYA LACINIOSA (Мех. f.) Loud. 4031. The large-fruited, shell-bark hickory is rare in our area, a few trees only having been noted along Spring River, a few miles west of Carthage. CARYA ARKANSANA Sargent. BETULACEAE COoRYLUS AMERICANA Май. 891, 926. CORYLUS ROSTRATA Ait. 3455. OSTRYA VIRGINIANA (Mill.) K. Koch. 32, 380, 3522. BETULA NIGRA L. 925, 1544. FAGACEAE QUERCUS ALBA L. 81, 705. QUERCUS STELLATA Wang. 25,964, 2145. QUERCUS MACROCARPA Michx. 74, 1537. QUERCUS MUHLENBERGII Engelm, 26, 963, 1071, 1139, 3854. QUERCUS RUBRA L. 79. QUERCUS PALUSTRIS Muench. 901, 1275, 1514, 1901, 902. Quercus Scuneck Britton. 5, 78, 903, 962. QUERCUS VELUTINA Lam. 4, 27, 53, 1976. 1Ттееѕ and shrubs 2:206. 1913. [ Vor. 8 372 ANNALS OF THE MISSOURI BOTANICAL GARDEN QUERCUS VELUTINA, Var. MISSOURIENSIS Sarg. 1125. (JUERCUS MARILANDICA Muench. 19, 24, 32, 704. ULMACEAE ULMUS FULVA Miehx. 90, 376, 3424. ULMUS americana L. 83, 379, 385, 1636, 3536. CELTIS OCCIDENTALIS L. 1130, 1538, 2675. CELTIS MISSISSIPPIENSIS Bose. 47, 1279, 1280, 1277, 1551, 2082, 1278, 3659. CELTIS GEORGIANA Small. 1182, 1494, 875, 3489. MORACEAE CANNABIS SATIVA L. 902. Нтгмтлл/в LuruLus L. 410, 1258. Нгмтл/0в JAPoNICUs Sieb. & лсе. 3799. MACLURA POMIFERA (Raf.) Sehneider. 1129, 3303. Morus RUBRA L. 54, 3355. Morus ALBA L. 901. Morus NIGRA L. 3729. URTICACEAE LAPORTEA CANADENSIS (L.) Gaud. 468. PILEA PUMILA (L.) Gray. 339, 3256. БВОЕНМЕНГА CYLINDRICA ( L.) Sw. 421, 830, 876, 3446. PARIETARIA PENNSYLVANICA Muhl. 346. SANTALACEAE CoMANDRA UMBELLATA (L.) Nutt. 582, 3607, 3626. CoMANDRA PALLIDA À. DC, 2460, 3774. ARISTOLOCHIACEAE ASARUM CANADENSE L. 693, 922. ARISTOLOCHIA TOMENTOSA Sims. 29, 948. ARISTOLOCHIA SERPENTARIA L. 1552, 1915, 1986, 2227, 2422, 2259. POLYGONACEAE Rumex cnisPUS L. 151, 152. Комех aLtisstmus Wood. 148, 149. RuMEX VERTICILLATUS L. 1282, 2314. Комех oBTUSIFOLIUS L. 153, 537, 2689. Rumex ACETOSELLA L. 150, 1079. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI POLYGONUM AVICULARE L. 252, 447, 482. POLYGONUM ERECTUM L. 894. PoLyGoNUM RAMOSISSIMUM Michx. 441, 1540. PorvcoNuM TENUE Michx. 2775, 2847. PoLYGONUM LAPATHIFOLIUM L. 250. Potyconum MvunrzxsBznor (Meisn.) Wats. 2335. POLYGONUM PENNSYLVANICUM L. 249, 442, 443, 444, 3882. POLYGONUM LONGISTYLUM Small. 1078. Potyconum HYDROPIPER L., 251. Ротусоктм acre НВК. 445. POLYGONUM ACRE, Var. LEPTOSTACHYUM Meisn. 154, 2720. POLYGONUM ORIENTALE L. 2593. Potyconum PERSICARIA L. 155, 446. 373 PoLYGONUM HYDROPIPEROIDES Michx. 464, 2287, 2332, 2438, 9779. POLYGONUM VIRGINIANUM L. 156, 3210. POLYGONUM SAGITTATUM L. 790, 3053, 3261. POLYGONUM CONVOLVULUS L. 248. POLYGONUM SCANDENS L. 611. CHENOPODIACEAE CYCLOLOMA ATRIPLICIFOLIUM (Spreng.) Coult. 3841. CHENOPODIUM AMBROSIOIDES L, 812, 1043, 1549. CHENOPODIUM ANTHELMINTICUM L. 2923, 3442, 3846. CHENOPODIUM HYBRIDUM L. 811, 3804, 3837. CHENOPODIUM ALBUM L. 1040, 2041, 2357, 3838. CHENOPODIUM BERLANDIERI Moq. 1042. CHENOPODIUM MURALE L. 3106, 3419. CHENOPODIUM Вовстахтум Мод. 787. CHENOPODIUM LEPTOPHYLLUM Nutt. 1041. CHENOPODIUM LEPTOPHYLLUM, Var. OBLONGIFOLIUM 5. Wats. 349. AMARANTHACEAE AMARANTHUS RETROFLEXUS L. 427, 2624, 3114. AMARANTHUS HYBRIDUS L. 2827. AMARANTHUS GRAECIZANS L, 1038. AMARANTHUS BLITOIDES Š. Wats. 615. AMARANTHUS SPINOSUS L. 345. ACNIDA TAMARISCINA (Nutt.) Wood. 425, 1302, 1560. IRESINE PANICULATA (L.) Ktze. 783, 1145, 1483. [Vor. 3 374 ANNALS OF THE MISSOURI BOTANICAL GARDEN FROELICHIA GRACILIS Moq. 3227. Sarsora Kaur L PHYTOLACCACEAE PHYTOLACCA DECANDRA L. 357. NYCTAGINACEAE OXYBAPHUS NYCTAGINEUS (Michx.) Sweet. 436, 484, 3415. OXYBAPHUS FLORIBUNDUS Chois. 871, 1487, 3079. OXYBAPHUS ALBIDUS ( Walt.) Sweet. 1096, 2761, 3135, 3651, 3480. ILLECEBRACEAE ANYCHIA POLYGONOIDES Raf. 248, 1316, 1559. ANYCHIA CANADENSIS (L.) BSP. 423, 938. AIZOACEAE MOLLUGO vERTICILLATA L. 341, 3886. СЕОСАВРОХ MINIMUM Mack. 3921, 5517. This interesting plant has so far been found only in one spot, in sandstone barrens on the high prairies, four miles north of Alba. In connection with his description of it? Mr. Kenneth K. Mackenzie says: “This plant is probably to be referred to the family Aizoaceae, or as treated in the Synoptical Flora I: 256 the Ficoideae, and to the tribe Aizoideae of that family. In many respects it seems to come closer to the genus Cypselea than to any other North American genus. It differs markedly in the absence of stipules and style and in the eapsule not being circumscissile, The other genera of the tribe in question, found in this country, are succulent plants with circumscissile capsules and cornute calyx-lobes. «The tribe Mollugineae of the same family characterized by a calyx divided nearly or quite to the base, and represented in the United States by two genera having 3-celled ovaries, is less closely related to our plant. Nor can our plant be considered an apetalous representative of the Alsinaceae, as the sepals in that family are distinct or very nearly so. It seems in fact to represent a well-characterized genus."' 1Torreya 14:67. 1914. 1916] PALMER—PLANTS 0F JASPER COUNTY, MISSOURI 375 CARYOPHYLLACEAE SPERGULA ARVENSIS L. 3711. SAGINA DECUMBENS (Ell.) T. & G. 912. ARENARIA SERPYLLIFOLIA L. 1105. ARENARIA PATULA Michx. 506, 1245. STELLARIA MEDIA (L.) Cyrill. 504, 1752. CERASTIUM VULGATUM L. 562, 3634. CERASTIUM VISCOSUM L. 3272. CERASTIUM BRACHYPODUM (Engelm.) Robinson. 302, 568. CERASTIUM nutans Raf. 570, 1680, 3356. AGROSTEMMA QITHAGO L. 517, 2093. SILENE ANTIRRHINA L. 413, 749, 889. SILENE REGIA Sims. 278, 656. SILENE STELLATA (L.) Ait. f. 342. SILENE NocTIFLORA L. 3974. SAPONARIA OFFICINALIS L. 435. PORTULACACEAE CLAYTONIA VIRGINICA L. 365. A form of this plant growing in low rich woods has leaves much broader than in the prairie plants, sometimes 1.5 cm. broad. TALINUM PARVIFLORUM Nutt. 2268. TALINUM CALYCINUM Engelm. 909, 2269. PORTULACA OLERACEA L. 820. Ровттл,Аса NEGLECTA Mack. & Bush. 3108. PonruLaca PILoSA L. 908, 1412. CERATOPHYLLACEAE CERATOPHYLLUM DEMERSUM L. 1438, 1815. NYMPHAEACEAE NYMPHAEA ADVENA Ait. 888, 1917. RANUNCULACEAE RANUNCULUS AQUATALIS, VAT. CAPILLACEUS DC. 2028, 2313, 2320, RANUNGULUS OBLONGIFOLIUS Ell. 1961, 1965, 2037, 3055. RANUNCULUS MICRANTHUS Nutt. 571, 1661, 1665. RANUNCULUS ABORTIVUS L. 543, 752, 3650. 376 ANNALS OF THE MISSOURI BOTANICAL GARDEN RANUNCULUS RECURVATUS Poir. 911, 1893, 3777. RANUNCULUS FASCICULARIS Muhl. 145, 1607. RANUNCULUS SEPTENTRIONALIS Poir. 913, 1634. RANUNCULUS HISPIDUS Michx. 912, 1894. MYOSURUS MINIMUS L. 1630, 1660, 3390. THALICTRUM DASYCARPUM Fisch, & Lall. 2030. THALICTRUM REVOLUTUM DC. 496, 2110. ANEMONELLA THALICTROIDES (L.) Spach. 143, 144. ANEMONE CAROLINIANA Walt. 141, 142, 1765. ANEMONE VIRGINIANA L. 491, 915. CLEMATIS Prronert Т. & G. 697, 698, 1101, 2326. CLEMATIS MISSOURIENSIS Rydb. 3690. IsoevnvM BITERNATUM (Raf.) T. & G. 1618, 1761. AQUILEGIA COCCINEA Small. 587. DELPHINIUM TRICORNE Michx. 147. Пкімрніхітум Penarpr Huth. 146, 1864. CIMICIFUGA RACEMOSA (L.) Nutt. 869, 1102, 2325. ANONACEAE ASIMINA TRILOBA Dunal. 31, 37, 813. MENISPERMACEAE Coccurvs caRoLINUS (L.) DC. 614, 894, 2377, 3429. MENISPERMUM CANADENSE L. 143, 2054. Catycocarpum Lyoni (Pursh) Nutt. 735, 3236. BERBERIDACEAE PopoPHYLLUM PELTATUM L. 692, 3601. CAULOPHYLLUM THALICTROIDES (L.) Michx. 3944. LAURACEAE SASSAFRAS VARIIFOLIUM (Salisb.) Ktze. 8, 34, 709, 710. BENZOIN AESTIVALE (L.) Nees. 20, 35, 383, 344, 708. PAPAVERACEAE SANGUINARIA DILLENIANA Greene. 1107, 1137, 1610. FUMARIACEAE Dicentra CUCULLARIA (L.) Bernh. 290, 1635. Совурліля FLAVULA (Raf.) DC. 289, 1616, 1676. CORYDALIS CRYSTALLINA Engelm. 288, 888, 3412. [Vor. 3 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI CoryDALIS AUREA Willd. 1625, 1678, 1702. CoryDALIS MONTANA Engelm. 1645, 1674. CRUCIFERAE DRABA CAROLINIANA Walt. 282, 1600, 1608. DRABA CUNEIFOLIA Nutt. 1615, 1673. DRABA BRACHYCARPA Nutt. 285, 1609. LEPIDIUM VIRGINICUM L. 943. LEPIDIUM APETALUM Willd. 287, 2161. CAPSELLA Bursa-pastoris (L.) Medic. 283, 286, 1286, 3645. CAMELINA MICROCARPA Andrz, 2042. Brassica ALBA (L.) Boiss. 2553. Brassica ARVENSIS (L.) Ktze. 540, 2458. Brassica JAPONICA Siebold. 2346. Brassica NIGRA (L.) Koch. 2131. Brassica CAMPESTRIS L. (6049 of B. F. Bush). SISYMBRIUM OFFICINALE (L.) Scop. 459. 377 RADICULA NASTURTIUM-AQUATICUM (L.) Britten & Rendle. 940, 3282. RADICULA SESSILIFLORA ( Nutt.) Greene. 1911, 1980, 3250, 3301. RADICULA PALUSTRIS (L.) Moench. 525, 3009. Rapicuna aquatica (bat Robinson. 1285, 2184, 2486. BaRBAREA VULGARIS R. Br. 1863. SELENIA AUREA Nutt. 942, 1603, 1799. JopDANTHUS PINNATIFIDUS (Michx.) Steud. 405. DENTARIA LACINIATA Muhl. 284, 682, 1284, 1662, 3337. CARDAMINE BULBOSA (Schreb.) BSP. 1705, 1895. CARDAMINE ROTUNDIFOLIA Michx. 844. What appears to be this species has been found in a spring, near Spring River, four miles east of Carthage. CARDAMINE PARVIFLORA L. 755, 3629. CARDAMINE ARENICOLA Britton. 508, 1709, 1824, 3672. CARDAMINE PENNSYLVANICA Muhl. 3615, 3657. ARABIS DENTATA Т, & G. 573, 1749. ARABIS VIRGINICA (L.) Trel. 336, 757, 1283, 1599. ARABIS HIRSUTA (L.) Scop. 941, 2535, 2960, 3701. ARABIS LAEVIGATA (Muhl.) Poir. 486, 761. ARABIS CANADENSIS L. 497, 842, 3689. [Vor. 3 318 ANNALS OF THE MISSOURI BOTANICAL GARDEN SOPHIA INTERMEDIA Rydb. 1149, 1629, 1640, 1732, 1804, 843, 841. SOPHIA PINNATA ( Walt.) Britton. 1147, 1703, 1739, 3660. CAPPARIDACEAE POLANISIA GRAVEOLENS Raf. 2773. CLEOME SPINOSA L. 870. CRASSULACEAE PENTHORUM SEDOIDES L. 610, 784, 2721. Sepum NurTALLIANUM Raf. 872. SEDUM PULCHELLUM Michx. 350. SEDUM Nevi Gray. 3199, 3205. SAXIFRAGACEAE HEUCHERA nrnsUTICAULIS (Wheelock) Rydb. 530, 1108, 2536, 3654 Rings MISSOURIENSIS Nutt, 373, 575, 2009, 3596, 2373. PLATANACEAE PLATANUS OCCIDENTALIS L. 23, 1841. ROSACEAE OPULASTER INTERMEDIUS Rydb. 6, 10, 1455. GILLENIA STIPULATA ( Muhl.) Trel. 351, 1091, 1132, 3754. Pyrus roeNsis (Wood) Bailey. 105, 391, 1554, 3348, 3476, 3595. Pyrus roexsis, var. Рагменг Rehder. 2605, 3347, 3473. Pyrus Marus L. 467, 1722 AMELANCHIER CANADENSIS (L.) Medic. 2, 27, 955, 1603, 3658, 1269, 1558. CRATAEGUS STRONGYLOPHYLLA Sarg. 1968, 1973, 1983, 2752, 2759, 2786. CRATAEGUS TANTULA Sarg, CRATAEGUS FEROX Sarg. 905, 916, 1579, 1580, 1581, 1582, 1583, 1584, 1585, 1586, 1587, 1588, 1870, 1984. CRATAEGUS PALMERI Sarg. 1564, 1565, 1566, 1567, 1568, 1569, 1570, 1571, 1572, 1573, 1574, 1575, 1576, 1577, 3617. CRATAEGUS ROTUNDA Sarg. 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1472, 1979. CRATAEGUS PARCIFLORA Sarg. 1292, 1293. CRATAEGUS JASPERENSIS Sarg. 1290, 1291, 1578, 1579, 2758, 2869, 2881, 1886. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 379 CRATAEGUS RUBRISEPALA Sarg. 1284, 1285, 1301, 2700. CRATAEGUS MUNITA Sarg. 1468, 2006, 2728. CRATAEGUS PARADOXA Sarg. 1899, 1977, 1930, 1933, 1978, 2762, 2763, 2770, 2771, 2772. CRATAEGUS RUBRIFOLIA Sarg. 2286, 1287, 1288, 1289, 1301, 1464, 1887, 1971, 2868. CRATAEGUS VICINA Sarg. 1232, 1263, 1265, 1266, 1230, 1231, 1264, 1463, 1786, 2886, 2887, 2888, 2889, 2890. CRATAEGUS MACROPODA Sarg. 1227, 1228, 1229, 1257, 1258, 1259, 1260, 1261, 1262, 1263, 2736. CRATAEGUS SECTA Sarg, 1225, 1226, 1255, 1256, 1467, 1728, 1731, 2656, 2658, 2838. CRATAEGUS FURCATA Sarg. 1233, 1234, 1267, 1268, 1270, 1271, 1272, 1832, 1869, 2884. CRATAEGUS LUTENSIS Sarg. 2883. CRATAEGUS OVATA Sarg. 1959. CRATAEGUS BRACTEATA Sarg. 1253, 1254, 1274, 1828, 1842, 1885, 1876, 1469, 1474, 2603, 2755. CRATAEGUS ASPERA Sarg. 1245, 1246, 1839, 2755, 2757. CRATAEGUS DISJUNCTA Sarg. CRATAEGUS MAGNIFOLIA Sarg. 1247, 1248, 1249, 1250, 1251, 1252 1469, 1831, 1648, 1867, 1872, 1890, 2799, 2800. CRATAEGUS LASIANTHA Sarg. 1242, 1273, 1283, 1473, 1744, 2671, 2772, 2794, 2707, 1757. CRATAEGUS DUMETOSA Sarg, 1462, 1470, 1692. CRATAEGUS MOLLIS (Т. & G.) Scheele. 1282, 1700, 1746, 1748, 2715. CRATAEGUS LANUGINOSA Sarg. 1235, 1237, 1238, 1239, 1240, 1241, 1243, 1273, 1811, 1814, 1843, 1782, 3618. CRATAEGUS DASYPHYLLA Sarg. 1712, 1768. CRATAEGUS SPECIOSA Sarg, 1219, 1220, 1221, 1222, 1223, 1224, 1243, 1244, 1777, 1780, 1801, 1807, 2650, 2738, 2740, 2858, 2862, 2873, 2874, 2875, 2876, 2973. CRATAEGUS HISPIDULA Sarg. 1276, 1277, 1465, 1471, 2032, 2084, 2744, 2073, 2709. CRATAEGUS OBSCURA Sarg. 1271, 1278, 1279, 2031, 2052, 2069, 2072, 2619, 2632, 2711, 3858. ` [Vor. 3 380 ANNALS OF THE MISSOURI BOTANICAL GARDEN CRATAEGUS SPINULOSA Sarg. 1280, 1281, 2020, 2035, 2691, 2705, 2748, 3496. CRATAEGUS INSPERATA Sarg. 1274, 1275, 1466, 2039, 2063, 2064, 2777, 2863. CRATAEGUS MOLLICULA Sarg. 2036, 2037, 2055, 2749. CRATAEGUS SIMULATA Sarg. 1999, 2000, 2001, 2019, 2034, 2690, 2743, 2745. CRATAEGUS SPATHULATA, Var, FLAVANTHA Sarg. 1994, 2856, 2870, 2 Only one tree of this species has been found in a thicket near Joplin. This is the most northerly station recorded for this southern red haw, which is very common in some of the gulf states. FRAGARIA VIRGINIANA Duch. 308. FRAGARIA VIRGINIANA, Var. ILLINOENSIS (Prince) Gray. 1272. POTENTILLA MONSPELIENSIS L. 736, 803, 1985, 2113, 2411, 3758. PorENTILLA CANADENSIS L. 307, 907, 2479. A slender form found in the northwestern part of the county, with long filiform runners and under surface of leaves somewhat silvery, may be distinet, GEUM CANADENSE Jacq. 497, 2190, 2414. GEUM VERNUM (Raf.) T. & G. 352, 544, 3368. Ковоѕ occipENTAUIS L. 740, 3454, 3497. RUBUS ALLEGHENIENSIS Porter. 1270. Ковов Anprewsianus Blanchard. 99, 2025, 3432, 3453. RUBUS procuMBENS Muhl. 305, 475, 1271, 2229, 2234, 3422, 3435. RUBUS RUBRISETUS Rydb. 1594. AGRIMONIA STRIATA Michx. 3126. AGRIMONIA MOLLIS (Т. & G.) Britton. 306. AGRIMONIA PARVIFLORA Ait. 609, 1323. Rosa sETIGERA Michx. 33, 819, 2221, 2290. Rosa HELIOPHILA Greene. 50, 961, 2057, 2066, 2077, 2098, 2105, 2137, 2228. Rosa Моопви Lindl. 2051, 2132, 2840, 3406, 3438, 3440. Rosa RUBIGINOSA L, 3187. Rosa номплв Marsh. 98, 1274, 3775. RosA RUDIUSCULA Greene. 3715, 3749. Prunus вЕкоттхА Ehrh. 12, 954, 3377. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 381 Prunus ANGUSTIFOLIA Marsh. 2824, 3309. Prunus MusNsoNIANA Wight. & Hedrick. 3100, 3310, 3318, 3334, 3023, 3098. Prunus Manarzs L. 3149, 3378. Prunus PALMERI Sarg. 1677, 2233, 3525, 3904. Prunus HoRTULANA Bailey. 44, 821, 822, 823, 958, 825, 3328, 3340, 3586. PRUNUS HORTULANA, var. PUBENS Sarg. 2164, 2893, 2897, 3045, 3073, 2901. Prunus Persica (L.) Stokes. 2419. PRUNUS LANATA (Sudw.) Mack. & Bush. 45. To this species are doubtfully referred plums of the Ameri- cana group, common on prairies and in thickets and open woods. It is usually a shrub suckering freely and forming small thickets, producing small red acid fruit with a glaucous bloom, Occasionally it becomes a tree of considerable size. The true position of the species is still unsettled. LEGUMINOSAE DESMANTHUS ILLINOENSIS (Michx.) MacM. 431. ScHRANKIA UNCINATA Willd. 531, 3457. GYMNOCLADUS DIOICA (L.) Koch. 3, 3755. GLEDITSIA TRIACANTHOS L. 59, 729, 1541, 2186. Cassta Mepsceri Shafer. 126, 3441. Cassia CHAMAECRISTA L. 124, 2643. Cassia NICTITANS L. 123, 3164, 3471. CERCIS CANADENSIS L. 49, 387. BAPTISIA BRACTEATA (Muhl.) Ell. 728. BAPTISIA LEUCANTHA Т. & С. 568, 2464. BAPTISIA AUSTRALIS (L.) R. Br. 127. CROTALARIA SAGITTALIS L. 440, 840. TRIFOLIUM ARVENSE L. 3785. TRIFOLIUM PRATENSE L. 686. TRIFOLIUM REFLEXUM L. 500, 551, 1967, 3968. TRIFOLIUM STOLONIFERUM Muhl. 3967. TRIFOLIUM REPENS L. 1064. TRIFOLIUM HYBRIDUM L. 1067, 2058. TRIFOLIUM CAROLINIANUM Michx. 1789, 2058. [Vor. 3 382 ANNALS OF THE MISSOURI BOTANICAL GARDEN TRIFOLIUM PROCUMBENS L. 1065, 3040. MELILOTUS OFFICINALIS (L.) Lam. MELILOTUS ALBA Desr. 492, 2230. МЕПТСАСО SATIVA L. 2304. Hosackra AMERICANA (Nutt.) Piper. 595, 2597, 2968, 3877. PsoRALEA PEDUNCULATA (МШ.) Vail. 138. PsoRALEA TENUIFLORA Pursh. 139, 3367. AMORPHA CANESCENS Pursh. 20, 27. AMORPHA FRUTICOSA L. 15, 830. PETALOSTEMUM PURPUREUM ( Vent.) Rydb. 2983, 3487. PETALOSTEMUM PURPUREUM, Var. PUBESCENS Gray. 202. PETALOSTEMUM CANDIDUM Michx. 201, 1066, 3466. TEPHROSIA VIRGINIANA (L.) Pers. 147, 2308, 3125. RoBINIA Pseupo-Acacta L. 886. ASTRAGALUS CARYOCARPUS Ker. 399, 1290. ASTRAGALUS MEXICANUS A. DC. 3332. ASTRAGALUS CANADENSIS L. 885. ASTRAGALUS DISTORTUS 'T. & G. 588, 1704, 1798, 1829, 1932, 1956. DESMODIUM NUDIFLORUM (L.) DC, 2350, 2542, 2594. DESMODIUM GRANDIFLORUM ( Walt.) DC. 129, 367, 2898. DESMODIUM PAUCIFLORUM (Nutt.) DC. 2585, 2595. DESMODIUM ROTUNDIFOLIUM (Michx.) DC. 3059, 3121, 3160. DesmMopium CANESCENS (L.) DC. 419, 1067, 1068, 2539, 2589, 2615, 2697, 2687, 2696. DzswoprvM BRACTEOSUM (Місһх.) DC. 2695, 3204. DESMODIUM ILLINOENSE Gray. 463, 2280. Desmopium Оплехп Darl. 765, 2676, 2678, 2694, 2751. DESMODIUM PANICULATUM, var. PUBENS Т. & G. 2641, 2680, 2704. DESMODIUM CANADENSE (L.) DC. 2572. Певмортом SESSILIFOLIUM (Torr.) T. & G. 2582, 2681, 2760. DESMODIUM MARILANDICUM (L.) DC. 891, 2679, 2774. LESPEDEZA REPENS (L.) Bart. 887. LESPEDEZA VIOLACEA (L.) Pers. 2, 135, 3205, 3469. LESPEDEZA PRATREA (Mack. & Bush) Britton. 890, 3848. LespepEza Stuvet Nutt. 1062, 1063, 2826, 3468. LrsPEDEZA VIRGINICA (L.) Britton. 133, 134, 771, 889, 3867. LrsPEDEZA HIRTA (L.) Hornem. 888. LzsPEDEZA CAPITATA Michx. 3464, 3847. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 883 LESPEDEZA CAPITATA, Var, SERICEA H. & A. 140, 3458. LzsPEDEZA STRIATA (Thunb.) H. 6 А. 600. STYLOSANTHES BIFLORA (L.) BSP. 131, 132, 461, 2253. VICIA VILLOSA Roth. 2281. LATHYRUS PUSILLUS Ell. 2004. Aptos TUBEROSA Moench. 3022. PHASEOLUS PoLvsTACHYUS (L.) BSP. 5218. STROPHOSTYLES HELVOLA (L.) Britton. 778, 3876. STROPHOSTYLES UMBELLATA (Muhl.) Britton. 892. SrRoPHOSTYLES PAUCIFLORA (Benth.) Wats. 817, 2522, 3070. AMPHICARPA MONOICA (L.) Ell. 773. Ampuicarpa Рітснен Т. & G. 3051, 3194. GArACTIA VOLUBILIS (L.) Britton, 436. GALACTIA VOLUBILIS, Var. MISSISSIPPIENSIS Vail, 2833. LINACEAE LINUM vsrTATISSIMUM L. 899, 2543. LINUM svLCATUM Riddell. 264 LINUM MEDIUM (Planch.) Britton. 898, 1534, 2220, 2261. OXALIDACEAE OXALIS VIOLACEA L. 259. OXALIS stricta L. 1106, 2049. OXALIS CORNICULATA L. 260, 2965. OxALIS REPENS Thunb, 681, 1558. OXALIS INTERIOR Small, 433, 792, 877, 2556. GERANIACEAE GERANIUM MACULATUM L. 680, 756. GERANIUM CAROLINIANUM L. 268, 2088. RUTACEAE ZANTHOXYLUM AMERICANUM Mill. 64, 392, 718, 3529. PTELEA TRIFOLIATA L. 24, 389, 707, 1516, 2263, 3801. SIMARUBACEAE AILANTHUS GLANDULOSA Desf. POLYGALACEAE PoLYGALA INCARNATA L. 187, 2294, 2435. PoLYGALA SANGUINEA L. 188, 358, 2138, 2218, 2277. POLYGALA VERTICILLATA L. 823, 2367, 3763. [Vor. 3 384 ANNALS OF THE MISSOURI BOTANICAL GARDEN EUPHORBIACEAE Свотох GLANDULOSUS L. 451, 1539. Croton cAPITATUS Michx. 169, 170. Свотох MONANTHOGYNUS Michx. 166, 167. CROTONOPSIS LINEARIS Michx. 612, 927, 1317, 3240. ACALYPHA VIRGINICA L. 791. ACALYPHA GRACILENS Gray. 364, 438. TRAGIA NEPETAEFOLIA Cav. 1111, 2986, 453. TRAGIA RAMOSA Torr. 160. This species of the stinging spurge appears to be quite distinet from T. nepetaefolia in the floral characters and in the stiff erect habit of growth. The latter is usually a more or less twining vine, sometimes 5 m. high or more. PHYLLANTHUS CAROLINIENSIS Walt. 449, 1281, 3283. EUPHORBIA Presi Guss. 763, 397, 448, 813, 2719. EUPHORBIA MACULATA L. 450, 452, 926, 2718. EUPHORBIA MARGINATA Pursh. 3072, 3127. EUPHORBIA COROLLATA L. 156, 164, 165. EUPHORBIA DENTATA Michx. 168, 834, 2374. EUPHORBIA HETEROPHYLLA L. 161, 162, 3123, 3219. EUPHORBIA MISSOURIENSIS (Norton) Small. 1922, 2139. CALLITRICHACEAE CALLITRICHE HETEROPHYLLA Pursh. 886, 1614, 1658, 1816, 3731. CALLITRICHE DEFLEXA, var. Austint (Engelm.) Hegelm. 1908, 3015, 3728, 3681. ANACARDIACEAE Ruvs GLABRA L. 75, 1128. Кнов coPALLINA L. 15, 959. Raus Toxicopenpron L. 52, 1517, 1903, 2135. Ruvs canavensis Marsh. 7, 1866. Кнев rRILOoBATA Nutt. 206, 873, 1109, 1900, 1805. This shrub, commonly known as polecat bush, is quite dis- tinet from Rhus canadensis. It is usually found on dry lime- stone ledges or rocky bluffs and is a stout shrub 1 to 1.5 m. high, with stems sometimes 4 or 5 em. in diameter. It flowers much later than R. canadensis, the leaves being more than half grown at the time of blooming. R. canadensis, a slender shrub, is one of the earliest flowering plants of spring. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 385 AQUIFOLIACEAE ILEX pecipua Walt. 1309, 1310, 1326, 1822. CELASTRACEAE EVONYMUS ATROPURPUREUS Jacq. 46, 1518. CELASTRUS SCANDENS L. 77, 712 STAPHYLEACEAE STAPHYLEA TRIFOLIA L. 66, 817, 818, 3531. ACERACEAE ACER SACCHARUM Marsh. 31, 384, 3671, 3532. ACER SACCHARINUM L. 21, 377, 381. Хкатхро ACEROIDES Moench. 11, 382, 923, 1686, 3519. The box elders differ much in foliage and to some extent in the fruit. There are two rather distinct forms in our region: one with glabrous twigs and leaves is the true Acer Negundo or Negundo Negundo, to adopt the more recent name; the form with broad rugose pubescent leaflets is N. texanum of Rydberg. The fruit in both these species is somewhat con- stricted into a stipe-like base. A third form with pubescent leaves and fruit not so constricted has been distinguished as N. interius Rydberg. SAPINDACEAE SAPINDUS DRUMMosNpi H. & А. 3984, 4020. The most northerly station recorded for this tree, popularly called soapberry or wild chinaberry, is along a small creek near Careytown, in the northern part of our county. It has been found at two other stations in the county, both of them on Center Creek. It grows along dry limestone bluffs having a southern exposure. "There are not many plants at either of these colonies and it is not unlikely that it may become extinct in our area, AESCULUS GLABRA Willd. 1681, 1685, 1758, 3530. BALSAMINACEAE IMPATIENS PALLIDA Nutt. 361. IMPATIENS BIFLORA Walt. 467. RHAMNACEAE RHAMNUS LANCEOLATA Pursh. 56, 70. [Vor. 3 386 ANNALS OF THE MISSOURI BOTANICAL GARDEN CEANOTHUS AMERICANUS L. 40, 813. CEANOTHUS OVATUS, Var. PUBESCENS Т, & G. 395, 2258. VITACEAE PSEDERA QUINQUEFOLIA (L.) Greene. 1265, 3842. PSEDERA QUINQUEFOLIA, var. HIRSUTA (Donn) Rehder. 1264. PsEDERA QUINQUEFOLIA, var. Батхт-Ратллт (Koehne & Grabner) Rehder. 80, 1496. Cissus AMPELOPsIS Pers, 62, 1589, 4002. VITIS CINEREA Engelm. 458, 2045, 2185, 2249, 2499. Vitis BIcoLoR Le Conte. 2822. VITIS CORDIFOLIA Miehx. 53, 67, 897, 3966. Vitis Ілхвесомп Buckley. 58, 1318, 2104, 2461, 2462, 2661, 3992. Vitis LINSECOMII, var. GLAUCA Munson. 2209, 2244, 2245, 2475, 4028. MALVACEAE Авотпох ТнкорнвАвтт Medic. 915, 2561. MALVASTRUM ANGUSTUM Gray. 437, 1547. SIDA sprnosa L. 596, 2470. MALVA ROTUNDIFOLIA L, 882. CALLIRHOE DIGITATA Nutt. 348, 3420. Hisiscus syriacus L. 3460. Нтвтвсев incanus Wendland. 1089, 1306, 1548. HIBISCUS MILITARIS Cav. 802, 1305. Hibiscus Trronum L. 839, 2600, 3450. HYPERICACEAE ASCYRUM HYPERICOIDES L. 538, 1670. HYPERICUM PUNCTATUM Lam. 2383, 2412. HYPERICUM PSEUDOMACULATUM Bush. 2921. HYPERICUM PROLIFICUM L. 7263, 1902, 3012. Hypericum CISTIFOLIUM Lam, 124, 175, 896. HYPERICUM MUTILUM L. 177, 1097. Hypericum Овсммохоп (Grev. & Hook.) T. & G. 262, 880, 1103, 3226, 3241. CISTACEAE HELIANTHEMUM Magus BSP. 523. 1916] ) PALMER—PLANTS ОҒ JASPER COUNTY, MISSOURI 387 LECHEA VILLOSA Ell. 796, 900, 1315. LECHEA TENUIFOLIA Michx. 266, 836. VIOLACEAE HYBANTHUS сохсогов (Forster) Spreng. 884, 1683, 1835, 3172. VIOLA PEDATA, Var. LINEARILOBA DC. VIOLA MISSOURIENSIS Greene. 1663, 1675, 1707, 1905, 2022, 3404, 3738, VIOLA PAPILIONACEA Pursh. 1639, 1686, 1957, 3267. VIOLA PALMATA, var. DILATATA ЕП, 1951. VIOLA TRILOBA Schwein. 1665, 1715, 3041, 3333, 3351, 3666, 3688, 8552, 3703. VIOLA SORORIA Willd, 845, 1110, 272, 1626, 1633, 1716, 846, 847, 1687, 2089, 3335. VIOLA FIMBRIATULA Sm. 3343, 3376. VIOLA БАСІТТАТА Ait. 1638, 1897, 3391, 3399, 3588, 3605, 3717. VIOLA EMARGINATA Le Conte. 760, 1766, 3066, 3587, 8717. VIOLA PEDATIFIDA С. Don. 561, 1644, 1764, 3346. VIOLA SCABRIUSCULA Sehwein, 271, 574, 1706, 1713, 1714, 737. VIOLA хтавтум Pollard. 1687. ViornA RAFINESQUII Greene, 673, 674, 1929. VIOLA PAPILIONACEA Ж TRILOBA. 2256, 3357, 3360, 3603, 3665. VIOLA PAPILIONACEA X PEDATIFIDA. 3331, 3345, 3393, 3425, 3593 3912. VIOLA SORORIA X TRILOBA. 565. VIOLA EMARGINATA X SORORIA. 1260, 3395. РА PASSIFLORACEAE PASSIFLORA LUTEA L. 141, 879, 2427. PASSIFLORA INCARNATA L. 6371. LOASACEAE MENTZELIA OLIGOSPERMA Nutt, 1099, 2356, 3004. CACTACEAE OPUNTIA HUMIFUSA Raf. 2142. OpuNTIA MACRORHIZA ? Engelm. 2278. LYTHRACEAE DriprPLIS DIANDRA (Nutt.) Wood. 1254, 3844. Котат,А RAMOSIOR (L.) Koehne. 800, 1476, 2419, 2626, 3246, 3821. [Vor. 3 388 ANNALS OF THE MISSOURI BOTANICAL GARDEN AMMANNIA COCCINEA Rottb. 2575, 4007. AMMANNIA AURICULATA Willd. 1104, 3248, 3822. LvrHRUM ALATUM Pursh. 267, 2337. CUPHEA PETIOLATA (L.) Koehne. 279, 2337. MELASTOMACEAE Кнехта LATIFOLIA Bush. 923, 949, 1256, 1907, 2433, 3049. ONAGRACEAE JUSSIAEA DIFFUSA Forsk. 910, 914, 2639. LUDVIGIA ALTERNIFOLIA L. 176, 801, 1243. LUDVIGIA PALUSTRIS ( L.) Ell. 1297, 2429, 3273, 3281, 3806. OENOTHERA MURICATA L. 416, 780. OENOTHERA MURICATA, var. CANESCENS (Т. & G.) Robinson. 2571. OENOTHERA BIENNIS L. 807. OENOTHERA LACINIATA Hill. 301, 3649, 3746. OENOTHERA LACINIATA, Var. GRANDIFLORA ( Wats.) Robinson. 550. OENOTHERA SPECIOSA Nutt. 480, 2083. OENOTHERA LINIFOLIA Nutt. 263, 2121, 3682. GAURA BIENNIs L, 112. Gaura Рітсневі (Т, & G.) Small. 102, 2549, 2640, 2660, 3113, 3147. GAURA PARVIFLORA Doug. 2494, 2591. CIRCAEA LUTETIANA L. 469, 881. HALORAGIDACEAE MYRIOPHYLLUM SCABRATUM Michx. 3242. MynIoPHYLLUM PROSERPINACOIDES Gill. 743, 3275. Established in a pond four miles northwest of Joplin. PnosERPINACA PALUSTRIS L. 3257. UMBELLIFERAE ERYNGIUM YUCOIFOLIUM Michx. 359, 1080. SANICULA GREGARIA Bicknell. 368. SANICULA CANADENSIS L. 2133, 2343. ERIGENIA BULBOSA ( Michx.) Nutt. 684, 759, 1598. CHABROPHYLLUM PROCUMBENS (L.) Crantz. 335, 572, 1619, 1812, (1559 of B. F. Bush). (HAEROPHYLLUM TEXANUM Соп. & Rose. 494, 1081, 2331. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 389 Osmoruiza CLAYTONI (Michx.) Clarke. 1892. OSMORHIZA LONGISTYLIS (Torr.) DC. 683, 1262. SPERMOLEPIS PATENS ( Nutt.) Robinson. 476, 530, 2310, 2345. SPERMOLEPIS ECHINATA (Nutt.) Heller. 2361. PTILIMNIUM NurraLum (DC.) Britton. 137, 2490. CICUTA MACULATA L. 172, 2491, 2557. CRYPTOTAENIA CANADENSIS (L.) DC. 490, 2134. ZIZIA AUREA (L.) Koch. 511, 1719. Zizia CORDATA (Walt.) DC. 1988. TAENIDIA INTEGERRIMA (L.) Drude. 121, 2087. EvLoPnus AMERICANUS Nutt. 489, 2210. CoNIUM MACULATUM Michx. 3707, 3997. T'HASPIUM BARBINODE ( Michx.) Nutt. 533, 1808, 2417. PorvrAENIA NurTrTALLII DC, 583, 685. PasTINACA SATIVA L. 2092. HERACLEUM LANATUM Michx. 1825, 2029. Torms Ахтнвівств (L.) Bernh. 2221, 2384, 3977. Daucus Canora L. 2371, 3191. Daucus PUsILLUs Michx. 539, 2270. CORNACEAE Cornus FLORIDA L. 63, 89, 814, 815, 816, 3632. Cornus Амомту/м Mill. 60, 375. CORNUS ASPERIFOLIA Michx. 57, 817, 1307, 2975, 2201. Cornus Вап,вүт Coult. & Evans. 1308, 2418. ERICACEAE МохотворА UNIFLORA L. 3155, 3254. The eurious and delieate Indian pipe or ghost plant has been found growing up through the humus in deep oak woods at several places in the county. MowornoPA Hyrorrrvs L. 2352. This species, known as pine sap, appears to be a great rarity, only a few plants having been found, June 27, 1909, on gravelly banks, growing with “reindeer moss," near Scotland Spring. VACOINIUM ARBOREUM Marsh. The tree huckleberry and the next species, the low bush huckleberry, barely get into our county in the southwestern [VoL. 3 390 ANNALS OF THE MISSOURI BOTANICAL GARDEN part. They are both abundant in the rocky woods bordering Shoal Creek, a little farther south in Newton County. VACCINIUM TENELLUM Ait. 2501. PRIMULACEAE ANDROSACE OCCIDENTALIS Pursh. 545, 696, 3584. SAMOLUS FLORIBUNDUS НВК. 1251, 2243, 2537, 2781. SrEIRONEMA CILIATUM (L.) Raf. 30, 738, 1090. STEIRONEMA LANCEOLATUM ( Walt.) Gray. 3887. ПорЕСАТНЕОХ MEADIA L. 585, 919, 920, 1726, 2702. CENTUNCULUS MINIMUS L. 3733. SAPOTACEAE BuMELIA LANUGINOSA ( Michx.) Pers. 28, 927, 2655. EBENACEAE DrosPvnos VIRGINIANA L. 62, 3974. There is mueh variation in the fruit and foliage of the persimmon. The common variety sometimes becomes a tree 10 or 12 m. high, forming small groves or thickets on upland prairies. The fruit is scarcely edible until after the first frosts. A not uncommon form, growing in similar situations, has large pubescent leaves with cordate bases which turn a bright yellow in autumn. Small trees producing very large soft pulpy fruit ripening early in September may belong to a distinet species. OLEACEAE FRAXINUS AMERICANA L. 924, 1542, 3533, 3636, 3667. FRAXINUS LANCEOLATA Borkh. 12, 117, 1513, 1776. FRAXINUS QUADRANGULATA Michx. 1124, 1515, 2080. ADELIA ACUMINATA Michx. 1249, 1627, 2074, 2188. APOCYNACEAE AMSONIA SALICIFOLIA Pursh. 296. 2488 This species grows usually along gr avelly branches, in large frutescent clumps 1 m. or more high. The leaves are narrower and the flowers smaller than in the next species, from which it appears quite distinct. А мвомтад TABERNAEMONTANA Walt. 921, 1819. APOCYNUM CANNABINUM L. 208, 1093, 1094, 3188. 1916] PALMER— PLANTS OF JASPER COUNTY, MISSOURI APOCYNUM PUBESCENS R. Br. 2336, 3426. APOCYNUM LEUCONEURON Greene. 3979, 4006. APOCYNUM ANDROSAEMIFOLIUM L. 2172. GENTIANACEAE SABATIA ANGULARIS (L.) Pursh. 405, 2506. SABATIA CAMPESTRIS Nutt. 363, 569, 2512. GENTIANA PUBERULA Michx. 344, 691, 3269. GENTIANA ANDREWSII Griseb. 1410, 1519, 3235, 3884. GENTIANA FLAVIDA Gray. 1592, 3128. ASCLEPIADACEAE ASCLEPIODORA VIRIDIS ( Walt.) Gray. 207. ASCLEPIAS TUBEROSA L. 211, 2820. ASCLEPIAS PURPURASCENS L. 213, 2235. ASCLEPIAS INCARNATA L. 2, 1536. ASCLEPIAS KANSANA Vail. 929. ASCLEPIAS AMPLEXICAULIS J. Е. Smith. 492, 946, 3035, 3234. ASCLEPIAS QUADRIFOLIA Jacq. 214, 3695. ASCLEPIAS VERTICILLATA L. 210, 819. ASCLEPIAS STENOPHYLLA Gray. 2309. ACERATES FLORIDANA ( Lam.) Hitehe. 209, 911. ACERATES VIRIDIFLORA ЕП, 1113. AGERATES VIRIDIFLORA, var. Ivrsi Britton. 212. GonoLosus LAEVIS Michx. 824, 1794, 3810. VINCETOXICUM CAROLINENSE (Jacq.) Britton. 206, 3956. VixcETOXICUM BALDWINIANUM (Sweet) Britton. 2416. CONVOLVULACEAE ТРОМОЕА COCOINEA L. 402, 1293. IPoMOEA HEDERACEA Jacq. 822, 3855. IPoMOEA PURPUREA (L.) Roth. 401, 821. IPOMOEA PANDURATA (L.) Meyer. 415, 1295. IPOMOEA LACUNOSA L. 768, 828, 2819. CONVOLVULUS sEPIUM L. 3075, 3136. CONVOLVULUS REPENS L. 3428. CoNVOLVULUS FRATERNIFLORUS Mack. & Bush. 3451. CONVOLVULUS ARVENSIS L. 523, 529. CUSCUTA oBTUsIFLORA НВК, 808, 1291, 1392, 3274. Cuscuta ARVENSIS Beyrich. 432, 3063. 391 [Vor. 3 392 ANNALS OF THE MISSOURI BOTANICAL GARDEN CUSCUTA CEPHALANTHI Engelm. 3197, 3835. Cuscuta Gronovir Willd. 2737, 3821. Cuscuta PARADOXA Raf. 818, 3069, 3129, 3861. POLEMONIACEAE Pnurox PILosA L. 503, 1546, 1742, 3957. PHLOX DIVARICATA L. 398, 557. PoLEMONIUM BEPTANS L. 581, 953, 1545. HYDROPHYLLACEAE HYDROPHYLLUM VIRGINIANUM L. 319, 2040, 2076. Kuuista NvorELEA L. 238, 1853. PHACELIA DUBIA (L.) Small. 261, 1879, 1888. PHACELIA HIRSUTA Nutt. 739, 2048. BORAGINACEAE HELIOTROPIUM TENELLUM (Nutt.) Torr. 439, 478, 1070, 2525, 3849, LAPPULA VIRGINIANA (L.) Greene. 422, 874, 2604, 3797. Муовоттв viretnica (L.) BSP. 507, 547, 3621, 3637. MYOSOTIS VIRGINICA, Var. MACROSPERMA (Engelm.) Fernald. 1906, 3569, 3663. LITHOSPERMUM CANESCENS ( Michx.) Lehm, 579, 1612, 3622. LrrHOSPERMUM ARVENSE L. 3406. LITHOSPERMUM ANGUSTIFOLIUM Michx. 580, 1590, 2370, 2381. ONOSMODIUM MOLLE Michx. 2050, 2140. ONOSMODIUM OCCIDENTALE Mack. 1069. ONOSMODIUM HISPIDISSIMUM Mack. 347, 472, 2303. VERBENACEAE VERBENA URTICAEFOLIA L. 197. VERBENA ANGUSTIFOLIA Michx. 200, 2981, 3519. VERBENA HASTATA L. 189, 1085, 1086, 2483. VERBENA STRICTA Vent. 199, 675, 2980. VERBENA BRACTEOSA Michx. 454. VERBENA CANADENSIS (L.) Britton. 304, 559. VERBENA ANGUSTIFOLIA X STRICTA 2932. VERBENA BRACTEOSA X STRICTA 601, 4025. LIPPIA LANCEOLATA Michx. 362, 2996, 3111, 3247, 3820. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI LABIATAE TEUCRIUM CANADENSE L. 277, 2541, 2568. IsANTHUS BRACHIATUS (L.) BSP. 934, 950, 3087. SCUTELLARIA LATERIFLORA L. 932, 2651 SCUTELLARIA INCANA Muhl. 8, 652. SCUTELLARIA CORDIFOLIA Muhl. 931, 2351, 2955, 2372. SCUTELLARIA PARVULA Michx. 505, 510, 516, 545, 2526. MARRUBIUM VULGARE L. 474. AGASTACHE NEPETOIDES (L.) Ktze. 772, 1058, 3095. Nepeta CATARIA L. 276. NEPETA HEDERACEA (L.) Trevisan. 1679, 2060, 3990. PRUNELLA VULGARIS L. 280. PHYSOSTEGIA VIRGINIANA (L.) Benth. 196. LAMIUM AMPLEXICAULE L. 1622, 1710. LAMIUM PURPUREUM L, 1753, 1989. Leonurus Carpraca L. 1060, 3782. STACHYS TENUIFOLIA Willd. 429, 598, 790, 3021. STACHYS LATIDENS Small. 2569. SALVIA Рттсневі Torr. 118, 1061. SALVIA LANCEAEFOLIA Poir. 1312, 3285. MONARDA FISTULOSA L. 3060. Moxanpa MOLLIS L. 203, 1059, 3043, (6045 of B. F. Bush.) MoxanpA BRADBURIANA Beck. 518, 2318. MONARDA CITRIODORA Cerv. 3800. BLEPHILIA CILIATA (L.) Raf. 2952. Brerna HIRSUTA (Pursh) Benth. 470, 3078. НЕгрЕома PULEGIOIDES (L.) Pers. 770. HEDEOoMA Hispipa Pursh. 515. НЕрЕоМА AcINOoIDEs Scheele. 2007. MELISSA OFFICINALIS L. 935. PYcNANTHEMUM FLEXUOSUM ( Walt.) BSP. 320, 2782. PYCNANTHEMUM PILOSUM Nutt. 317, 3809. CUNILA oniGANOIDES (L.) Britton. 3200. Lycopus RUBELLUs Moench, 933, 1311, 3883. Lycopus vIRGINICUS L. 3260. LYCOPUS MEMBRANACEUS Bicknell. 3843. Lycopus AMERICANUS Muhl. 424, 3857. MENTHA ROTUNDIFOLIA (L.) Huds. 1987, 3042. 393 394 ANNALS OF THE MISSOURI BOTANICAL GARDEN MENTHA SPICATA L. 1057, 2473, 2581. MENTHA PIPERITA L. 2024, 2331. MENTHA СТТВАТА Ehrh. 3185. MENTHA CANADENSIS, var. GLABRATA Benth. 2780, 3223. PERILLA FRUTESCENS (L.) Britton. 930. SOLANACEAE SoLANUM NIGRUM L. 257, 3109, 3208, 3220, 3227. SOLANUM CAROLINENSE L. 256. SoLANUM ELAEAGNIFOLIUM Cav. 918, 1324. SOLANUM nosrRATUM Dunal. 477. PHYSALIS ANGULATA L. 3093, 3221. PHYSALIS PUBESCENS L. 805. PHYSALIS PRUINOSA L. 2552, 3198. PHYSALIS BARBADENSIS Jacq. 2466. Рнүвлілв MissovuRIENSIS Mack. & Bush. 1252, 2988. Рнуватлв PUMILA Nutt. 253, 779, 1136, 2510, 2530. PHYSALIS HETEROPHYLLA Nees. 526, 917, 2081, 2511. PHYSALIS SUBGLABRATA Mack. & Bush. 255, 414, 904, 3110. PHYSALIS VIRGINIANA Mill. 254, 2008, 2500, 3776. Туситм HALIMIFOLIUM Mill. 1132. Datura Stramontum L. 2487. Datura Тато L. 258. Datura METEL L. 916. SCROPHULARIACEAE VrnBAsCUM THapsus L. 679. VERBASOUM BLATTARIA L. 269, 270, 4021. LINARIA VULGARIS Hill. 3044. LINARIA CANADENSIS (L.) Dumont. 896, 897, 1785. COLLINSIA VIOLACEA Nutt. 281, 947, 1767, 1823, 1856. SoROPHULARIA MARILANDICA L. 434, 1296. PENTSTEMON TUBIFLORUS Nutt. 90, 195. Рехтвтемох Птаттатлв (Sweet) Nutt. 194, 483, 937. MIMULUs RINGENS L. 1481. MIMULUS ALATUS Ait. 420, 951, 1075, 1550. Соховвл MULTIFIDA ( Michx.) Benth. 430. Bacopa ACUMINATA ( Walt.) Robinson. 799, 840, 2816. Bacopa ROTUNDIFOLIA (Michx.) Wettst. 338, 2383. [Vor. 3 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 395 ILYSANTHES DUBIA (L.) Barnhart. 1299, 3249. ILYSANTHES ANAGALLIDEA (Michx.) Robinson, 360, 1053, 1492. GRATIOLA VIRGINIANA L. 512, 549, 1992, 2124, 3685. GRATIOLA SPHAEROCARPA ЕП, 1966, 2176, 2297. Veronica VIRGINICA L. 11. VERONICA PEREGRINA L. 355, 3647, 3680. VERONICA ARVENSIS L. 363, 1806. DASISTOMA MACROPHYLLA (Nutt.) Raf. 418. AUREOLARIA GRANDIFLORA, var. CINEREA Pennell. 1, 10, 1298, 2616. ToMANTHERA AURICULATA ( Michx.) Raf. 613, 825. AGALINIS ASPERA (Doug. ) Britton. 2674. AGALINIS FASCICULATA (Ell) Raf. 604, 938, 1054, 1477, 3816; 3817, 3833, 3863. AGALINIS TENUIFOLIA (Vahl) Raf. 939, 833, 1140, 3132, 3148, 3818, 3862, 3875. AGALINIS BressEvANA Britton. 603, 1055, 2645, 3232, 3871, 3884. AGALINIS GarrTINGERI Small. 936, 1056, 1422, 2302, 3103, 3131, 3202, 3206. AGALINIS VIRIDIS (Small) Pennell. 816. BuCHNERA AMERICANA L. 108, 2441, 3064. CasTILLEJA COCCINEA (L.) Spreng. 502, 3408, 3639, 3640. PEDICULARIS CANADENSIS L. 549. LENTIBULARIACEAE UrricuLaRIA VULGARIS L. 2321, 3259. UTRICULARIA MINOR L. 3258. OROBANCHACEAE OROBANCHE UNIFLORA L. 703. BIGNONIACEAE Tecoma RADICANS (L.) Juss, 18. CATALPA SPECIOSA Warder. 884. CATALPA BIGNONIOIDES Walt. 2307. ACANTHACEAE DIANTHERA AMERICANA L. 297, 458. RUELLIA сплоѕА Pursh. 356, 1309. RvELLIA STREPENS L. 528, 1310. DIAPEDIUM BRACHIATUM (Pursh) Ktze. 599, 1482, 1591. [Vor. 3 396 ANNALS OF THE MISSOURI BOTANICAL GARDEN PHRYMACEAE PHRYMA LEPTOSTACHYA L. 455, 1098. PLANTAGINACEAE PLANTAGO MAJOR L. 2580. Ртахтасо Кост Dene, 157, 2349. PLANTAGO LANCEOLATA L. 159. PrANTAGO MEDIA L. 1854. PLANTAGO ARISTATA Michx. 235. PLANTAGO VrRGINICA L. 158, 1244, 1889, 1975. PLANTAGO ELONGATA Pursh. 546, 1790. RUBIACEAE GALIUM viRGATUM Nutt. 542, 2143. GALIUM APARINE L. 2025. GALIUM VAILLANTII DC. 353. GALIUM PILOSUM Ait. 336, 746, 747, 2248. GALIUM CIRCAEZANS Michx. 354. GALIUM TINCTORIUM L. 2180, 2208, 3745. GALIUM CONCINNUM Т, & С. 522, 1082, 2215, 3725. GALIUM TRIFLORUM Miehx. 1076, 2252, 2375. SPERMACOCE GLABRA Michx. 411, 782. DioptA rerrs Walt. 265, 273, 1255, 1825. CEPHALANTHUS OCCIDENTALIS L. 29, 396. НотвтомтА MINIMA Beck. 653, 654, 1133, 3543, 3619, 3643. HOUSTONIA PURPUREA L. 1220, 3011. CAPRIFOLIACEAE LONICERA SEMPERVIRENS L. 3186. Lonicera DioIcA L. 1912, 2078. Lonicera JAPONICA Thunb. 465, 607, 1077. SYMPHORICARPOS ORBICULATUS Moench. 97, 733, 734, 737, 796. TRIOSTEUM PERFOLIATUM L. 731, 732. VIBURNUM LENTAGO L. 9, 1556. VIBURNUM PRUNIFOLIUM L, 30, 378, 898, 1144, 3610. ViBURNUM RUFIDULUM Raf. 69, 394, 734, 1253. SAMBUCUS CANADENSIS L. 76, 1439, 3798. SAMBUCUS CANADENSIS, var. SUBMOLLIS f. ENGELMANNI Rehder. 3445, 3779. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 397 VALERIANACEAE VALERIANELLA RADIATA (L.) Dufr. 303, 1248, 1993. CUCURBITACEAE Srcvos ANGULATUS L. 597, 781. Ecurnocystis LOBATA (Michx.) T. & G. 878, 1259. CAMPANULACEAE SPECULARIA PERFOLIATA (L.) A. DC. 570, 671, 2148, 2158, 3962. SPECULARIA BIFLORA (R. & P.) Fisch, & Mey. 481. SPECULARIA LEPTOCARPA (Nutt.) Gray. 925, 2262, 2302, 3982. CAMPANULA AMERICANA L. 341, 672. LOBELIACEAE LoBELIA CARDINALIS L. 803, 910, 1134. LOBELIA SIPHILITICA L. 602. LOBELIA LzPTOSTACH YS А. DC. 190, 924. LoBELIA SPICATA, Var. HIRTELLA Gray. 191, 457, 2095. LOBELIA INFLATA L. 417, 466, 3802. COMPOSITAE CICHORIUM INTYBUS L. 3046. SERINIA OPPOSITIFOLIA ( Raf.) Ktze. 366. KRIGIA OCCIDENTALIS Nutt. 3936. ТК вїогА VIRGINICA (L.) Willd. 1883, 3668. Kricta DANDELIoN (L.) Nutt. 332, 1858. KRIGIA AMPLEXICAULIS Nutt. 655, 1891. TARAXACUM OFFICINALE Weber. 566, 3949. SONCHUS ARVENSIS L. 3239. бохснтув ASPER (L.) Hill 316, 2968. LACTUCA SCARIOLA L, 2463. LACTUCA SCARIOLA, var. INTEGRATA Gren. & Godr. 413, 1083, Lactuca CANADENSIS L, 904, 2698, 3130. LACTUCA SAGITTIFOLIA Ell. 412. LacTuca VILLOSA Jacq. 794. LACTUCA FLORIDANA (L.) Gaertn. 802. PRENANTHES ASPERA Michx. 312, 3176. PYRRHOPAPPUS CAROLINIANUS ( Walt.) DC. 329, 541, 2194. HIERAOIUM SCABRUM Michx. 3165. Hieracium Свохоуп L. 498, 2257, 3082, 3124, 3166. 398 ANNALS OF THE MISSOURI BOTANICAL GARDEN HIERACIUM LONGIPILUM Torr. 462, 2998, 4018. Iva сплата Willd. 785. AMBROSIA BIDENTATA Michx. 426. AMBROSIA TRIFIDA L. 224, 1141. AMBROSIA ARTEMISIAEFOLIA L. 428. AMBROSIA PSILOSTACHYA DC. 2559, 2663. XANTHIUM ECHINATUM Murr. 1303. XANTHIUM PENNSYLVANICUM Wallr. 766,767. XANTHIUM COMMUNE Britton. 1039, 1300. XANTHIUM GLABRATUM Britton. 1301, 1304. VERNONIA ORINITA Raf. 103, 2576, 2684, 3243, 3244. VERNONIA ALTISSIMA Nutt. 789. [Vor. 3 Vernonia Вагрутит Torr. 94, 2560, 2586, 2614, 2722, 2724, 2732. VERNONIA BALDWINI X CRINITA. 2654, 2724. ELEPHANTOPUS CAROLINIANUS Willd. 349. Есратовгум MaACULATUM L. 331, 4014. EUPATORIUM sEROTINUM Michx. 2683. EvPATORIUM ALTISSIMUM L. 17, 2642, 2685. EUPATORIUM PERFOLIATUM L. 863. EUPATORIUM URTICAEFOLIUM Reichard. 107, 3887. EUPATORIUM COELESTINUM Г. 326, 913, 3105. KUHNIA EUPATORIOIDES L. 328, 1035, 2731. LIATRIS SQUARROSA Willd. 860, 861, 1351, 3856. Laarris scariosa Willd. 112, 2849. LIATRIS PYCNOSTACHYA Michx. 1033, 1348. GRINDELIA LANCEOLATA Nutt. 181, 2578. AMPHIACHYRIS DRACUNCULOIDES ( DC.) Nutt. 113, 4033. Curysopsis PILOSA Nutt. 1346, 3026. SoLIDAGOo speciosa Nutt, 2823, 3872. SOLIDAGO RIGIDIUSCULA (T. & G.) Porter. 1012, 2677. SOLIDAGO ARGUTA Ait. 2688. SOLIDAGO ULMIFOLIA Muhl. 12, 916. SoLIDAGo RUGOSA Mill. 109, 110. SOLIDAGO GLABERRIMA Martens. 1010, 2565, 3068, 3988. SOLIDAGO NEMORALIS Ait. 2693, 2839, 3853, 3868. SOLIDAGO RADULA Nutt. 2831. SOLIDAGO ALTISSIMA L. 1006. SOLIDAGO ALTISSIMA, Var. PROCERA ( Ait.) Fernald. 1008, 1009. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 399 SOLIDAGO SEROTINA Ait. 1007, 1350. SOLIDAGO RIGIDA L. 111. SOLIDAGO PETIOLARIS Ait. 88, 325. ботлодсо Маври Britton. 1350, 2622, 2694, 3190, 3869. EvrTHAMIA GRAMINIFOLIA (L.) Nutt. 196. BoLToNIA asrEROIDES (L.) L'Her. 209, 220, 3853. BOLTONIA LATISQUAMA Gray. 3873. ASTER PALUDOSUS АП. 605, 1328, 3177. ASTER OBLONGIFOLIUS, Var. RIGIDULUS Gray. 907, 1336, 1337, Aster PATENS Ait. 1475, 3866, 1335. Автев ANOMALUS Engelm. 1338, 1339. Aster AZUREUS Lindl. 101, 914, 1331. ASTER SAGITTIFOLIUS Wedemeyer. 87, 324, 1332. Aster Овоммохри Lindl. 851, 3864. ASTER TURBINELLUS Lindl. 323, 1334, 3865. ASTER ERICOIDES L. 1329. ASTER ERICOIDES, Var. PILOSUS Porter. 1327, 1595, 2778. ASTER MULTIFLORUS Ait. 2814, 51, 1015, 1030. Aster Trapescanti L. 1015, 1326, 2846. ASTER PANICULATUS Lam. 852. ASTER SALICIFOLIUS Ait. 83, 106, 1014, 3885. ASTER LINARIIFOLIUS L. 865, 905. ERIGERON PULCHELLUS Michx. 564, 692, 824, 1341. ERIGERON PHILADELPHICUS L. 698, 1340, 1831, 1972. Ericeron annuus (L.) Pers. 754, 3965, 3954. Ентвввох RAMosus ( Walt.) BSP. 204, 1029, 2109, 3958, 3980. EmiaEROoN TENUIS T. & G. 1898, 2003. Енісевох wUDIFLORUS T. & G. 2097. PLUCHEA PETIOLATA Cass. 1511, 3880. ANTENNARIA OCCIDENTALIS Greene. 313. ANTENNARIA LONGIFOLIA Greene. 3901. There are probably two or three other species of this genus occurring in the county, the classification of which is not clear at present. GNAPHALIUM PURPUREUM L. 524, 1344, 3010. GNAPHALIUM OBTUSIFOLIUM L. 2725, 3201, POLYMNIA UVEDALIA L. 327, 866, 3159. [Vor. 3 400 ANNALS OF THE MISSOURI BOTANICAL GARDEN SILPHIUM LACINIATUM L. 183, 3439. SILPHIUM INTEGRIFOLIUM Michx, 857, 3459, 1024, 1345. SILPHIUM PERFOLIATUM L. 774, 3805. BERLANDIERA TEXANA DC. 2316. PARTHENIUM INTEGRIFOLIUM L. 536, 952. PARTHENIUM REPENS Eggert. 700, 701, 3961, 3978. Kourpta ALBA (L.) Hassk. 404, 3719. RUDBECKIA TRILOBA L. 1021, 3209, 3430. RvupBECKIA SUBTOMENTOSA Pursh. 867, 1022, 1347, 2415. RUDBECKIA HIRTA L. 182, 1020, 2319, 3788. RUDBECKIA speciosa Wenderoth. 3234. RUDBECKIA LACINIATA L. 1036. RUDBECKIA AMPLEXICAULIS Vahl. 2255. BRAUNERIA PALLIDA (Nutt.) Britton. 187. Lepacnys PINNATA ( Vent.) T. & G. 1031, 1032, 2563. HELIANTHUS ANNUUS L. 803, 2592, 2850. HELIANTHUS PETIOLARIS Nutt. 856. HELIANTHUS SCABERRIMUS Ell. 184, 1025. HELIANTHUS MOLLIS Lam. 186. HELIANTHUS GROSSESERRATUS Martens. 114, 1343. HELIANTHUS HIRSUTUS Raf. 407, 1023. HELIANTHUS HIRSUTUS, Var. TRACHYPHYLLUS T. & G. 1026. HELIANTHUS TUBEROSUS L. 2713. HELIANTHUS TUBEROSUS, Var. SUBCANESCENS Gray. 1478, 2659. ACTINOMERIS ALTERNIFOLIA (L.) DC. 322, 810, 3157. VERBESINA VIRGINICA L. 310, 321. VERBESINA HELIANTHOIDES Michx. 864, 3752. COREOPSIS TINCTORIA Nutt. 850, 2387, 2507. CorEOPSIS GRANDIFLORA Hogg. 179. Conzorsis PUBESCENS Ell. 370, 371, 806, 849. Corropsis PALMATA Nutt. 1030. Совкорв1в TRIPTERIS L. 298, 2584. BipENs FRONDOSA L. 853, 854, 855, 2692. BIDENS cERNUA L. 1019, 1479. BIDENS CERNUA, Var, ELLIPTICA Wieg. 1018. BIDENS BIPINNATA L. 309, 1017. BIDENS aristosa (Michx.) Britton. 1480. Bipens rNvoLvcRATA (Nutt.) Britton. 180. 1916] PALMER—PLANTS OF JASPER COUNTY, MISSOURI 401 MaRSHALLIA CAESPITOSA Nutt. 2015, 2153. HELENIUM NUDIFLORUM Nutt. 2315, 3233. HELENIUM AUTUMNALE L. 85, 1037, 3860. HELENIUM TENUIFOLIUM Nutt. 318, 3889. DvssoprA PAPPOSA ( Vent.) Hitche. 827, 3192. ACHILLEA MILLEFOLIUM L. 315. ANTHEMIS Cotuta L. 744. MATRICARIA SUAVEOLENS (Pursh) Buchenau. 2372, 4013. CHRYSANTHEMUM LEUCANTHEMUM L. 330, 3709, 3981. TANACETUM VULGARE, Var. INCISUM DC. 2548. ARTEMISIA LUDOVICIANA Nutt. 1027, 1028, 2747, 3473, 3874. ARTEMISIA MEXICANA Willd. 906. ARTEMISIA ANNUA L. 1034. ERECHTITES HIERACIFOLIA (L.) Raf. 903. CACALIA ATRIPLICIFOLIA L. 868, 2598. САСАТЛА TUBEROSA Nutt. 534. SENECIO OBOVATUS, var, RoTUNDUS Britton. 1834, 3521, 3669. SENECIO PLATTENSIS Nutt. 314, 3383, 3942. Arctium MINUS Bernh. 858. CIRSIUM LANCEOLATUM (L.) Hill. 2502. CIRSIUM DISCOLOR ( Muhl.) Spreng. 3058. CIRSIUM aALTISSIMUM (L.) Spreng. 189. LEPTILON CANADENSE (L.) Britton. 403,769. LzPTILON DIVARICATUM (Michx.) Raf. 1169. 859, 1342, 1667, Annals о Missouri Botanical Garden Vor. 3 NOVEMBER, 1916 No. 4 PISTILLARIA (SUBG. PISTILLINA) THAXTERI, BURT N. ӨР. THE SMALLEST KNown HyMENOMYCETE EDWARD ANGUS BURT Mycologist and Librarian to the Missouri Botanical Garden Associate Professor in the Henry Shaw School of Botany of Гази оп University On a recent visit to Ше Mycological Herbarium of Harvard University, I was given for study by Professor Thaxter a curious fungus, collected at West Haven, Connecticut, in 1888. This fungus is a hymenomycete of very simple structure and exceedingly minute size — so minute that the fructifications are not visible to the naked eye unless rendered so by special illumination and background, as in the case of the dust particles of the air becoming visible in a beam of sunlight thrown across a darkened room. By the aid of a lens the fructifications may be seen scat- tered on the surface of very rotten wood, merely gregarious, not united into clusters. One hundred and fifteen have been counted on an area 2 em. long by 4 em. broad. The fructifica- tions, after being kept twenty-eight years in the herbarium, are whitish to cartridge-buff throughout; each has a subglobose head, the pileus supported on a slender stem, and in its form suggests the sporangium of a minute myxomycete, such as a Physarum. In figs. 1 and 2 are shown two fructifications under magnification of 63 diameters; in fig. 1 these fructifica- tions were sketehed in dry condition, as they were on the ! [ssued January 12, 1917. ANN. Mo. Bor. GARD., Vor. 3, 1916 (403) [VoL. 3 404 ANNALS OF THE MISSOURI BOTANICAL GARDEN wood; in fig. 2 the same fructifications are shown after being removed from the wood and mounted in water. The structure of the fructification is shown by the higher magnification of fig. 3. From a layer of hyphae at or near the surface of the substratum, hyphae start out together at right angles to the substratum and are closely joined in a Pistillaria Zeie? 1, two fructifications in dried condition on wood, X , the same fructifications in an aqueous mount, x63; 3, Ae longitu bur optieal section of a fructification, X380; 4, hyphae, showing absence of clamp connections, X64 5, cluster ‘of young basidia, X640; 6, two База with sterigmata, X040; 7, five basidiospores, x 640. cylindric column about 60 u long and 20-40 шіп diameter in the dried specimens, swelling to 25—50, and rarely 80 y, in diameter when the specimens are wet, treated with potas- sium hydrate or lactic acid, and mounted in microscopical preparations. These hyphae are hyaline, thin-walled, about 12-2 р in diameter, and not incrusted nor nodose-septate (fig. 4). At the outer end of the stem the hyphae pass into the pileus which is distinguishable from the stem by its obversely conical form, as shown under a magnification of 380 diameters in fig. 3. The obconical form of the pileus is due to repeated branching of its hyphae as they extend directly from the stem to the surface of the pileus. The manner of branching and of increase in diameter of the pileus is shown in figs. 3 and 5. At the outer peripheral end the terminal cell of each hyphal branch becomes swollen with pro- 1916] BURT—PISTILLARIA THAXTERI 405 toplasmie contents and differentiates into a simple basidium somewhat clavate in form, 13-17 4-44 и, when fully mature, which bears four spores upon short sterigmata (figs. 5 and 6). The spores are hyaline, even, slightly flattened on one side, pointed at the base, 5-934 и (fig. 7). No cystidia, hairs, or organs other than basidia have been found in the hymenium. This fungus is remarkable not only for its minute size— and it is by far the smallest known species of the toadstool kind—but also for its extreme simplicity of structure. A few hyphae extend out together in a compact bundle from the vegetative mycelium, and at a little distance from the sub- stratum simply branch and terminate in basidia bearing the usual basidiospores. No additional accessory, supporting, or secretory organs of any kind are differentiated, nor is there any perceptible differentiation into cortical and medullary regions in the fructification, nor any curvature of the fertile hyphae so that the basidia will be directed towards special cavities or towards the substratum; on the contrary, the whole fructification is as simple as a sheaf of wheat. A few hyphae stand out together from the substratum — probably for mutual support—and produce as simply and directly as possible their complement of basidia and basidiospores, and form both distinct stem and pileus of the simplest possible structure. The primordium of the pileus in its ontogeny in more highly developed species is not simpler. Quelet1 published under the name Pistillina hyalina Quelet, n. gen. and sp. the description of a fungus closely related to the American species which I am describing. P. hyalina is ten times as large as our fungus, clearly visible to the naked eye, and has elongated, aculeate spores. Quelet’s genus Pis- tillina is regarded as a subgenus under Pistillaria of the Clavariaceae by Saccardo.2 While Pistillina appears to be a needed genus for such species as that for which it was founded and for the present American species, still the few species * Champ. Jura et Vosges, Suppl. 10, Assoc. Fr. Avanc. Sci. 9: 671. pl. 8. f. 12. 1880. ? Syll. Fung. 6: 759. 1888. [Vor. 3, 1916] 406 ANNALS OF THE MISSOURI BOTANICAL GARDEN which would be clearly comprehended by it would be con- nected with the usual elongated forms of Pistillaria by some intermediate species found in Europe, where the species of Pistillaria appear to be more numerous and more frequent than in North America. The present American species may be characterized as follows: Pistillaria (subgen. Pistillina) Thaxteri Burt, n. sp. Fructifications gregarious, pileate, erect, drying whitish to eartridge-buff; pileus hemispherical, puberulent, attenuated at the base into a cylindric stem composed of hyaline, thin- walled, even-walled, parallel hyphae about 13-2 шіп diameter, not nodose-septate, not incrusted; basidia simple, subclavate, 13-17<444 р, with four sterigmata; spores hyaline, even, flattened on one side, pointed at the base, 5-9X 34—44 и; no eystidia nor paraphyses. Fructification 100-110 џ high; pileus 50-110 д in diameter, 40-50 д long; stem about 60 д long, 20-50 y, rarely 80 u, in diameter. On rotten wood, West Haven, Connecticut, November 7, 1888, В. Thaxter, type (in Farlow Herb. and Мо. Bot. Gard. Herb., 5724). The fructifications are but a fraction of the size of those of any other species of the genus and not visible to the naked eye. A NOTE ON THE ADAPTABILITY OF THE FOLIN MICRO-KJELDAHL APPARATUS FOR PLANT WORK AVIS Formerly Research Assistant to the Missouri Botanical Garden There is frequent need in most botanical laboratories for the determination of small amounts of nitrogen. Recourse is usually had to the familiar Kjeldahl method—a method, however, which proves rather cumbersome for certain types of work. Within recent years Folin and Farmer! of the Harvard Medical School have modified the original Kjeldahl method to the end of determining small amounts of urea nitrogen. In their investigations they found the method approached the original in accuracy, while in economy of material to be analyzed, in reagents, and in time for determination, it was superior. The Folin modification has been given an extended trial in this laboratory, and with a few modifications has been found admirably adapted to many phases of plant work. It is espe- cially good for demonstration of proteolytic changes, since the determination of nitrogen in the different protein frac- tions can be readily made. The nitrogen content of minute plant sections or organs can be determined—as well as the effect of light, darkness, nutrition, disease, etc., upon the nitrogen content of various plant parts. The Е 18 also adapted for work with advanced classes in plant physiology, the apparatus being easily set up, and requiring but little desk space and no hood; at the same time it is inexpensive to install. Apparatus.—The essential parts of the apparatus are as follows: 1. Kjeldahl flasks of 100 or 200 ce. capacity; 2. Folin fume absorbers ;? 1 Folin, O., and Farmer, C. J. A new method for the determination of total nitrogen in urine. Jour. Biol. Chem. 11: 493-501. 1912. 2 These can be obtained at most laboratory supply houses. ANN. Mo. Bor. GARD., VOL. 3, 1916 (407) [Vor. 3 408 ANNALS OF THE MISSOURI BOTANICAL GARDEN 3. Micro-burners; 4. Ostwald pipettes of 1 and 2 cc. capacity; 9. Condensers of small size.! The fume absorption apparatus consists of two parts: (1) a piece of straight glass tubing with side arms, (2) the fume absorber proper. This latter is a 25-сс. pipette, one end of which is invaginated into the bulb, the other bent midway at a little more than right angles. The invaginated end sits into the neck of the digestion flask, while the other end fits into a side arm of the glass tubing. The latter, in turn, connects to the suction pump, by which the fumes are drawn off. Both the Kjeldahl flasks and the tubing are supported in the man- ner illustrated in pl. 7. In the Folin modification, Jena test-tubes (200x:25 mm.) are used in digestion. In this laboratory the small Kjeldahl flask has been found to be better adapted to plant material, beenuse of the relatively high percentage of carbohydrates present, and the tendency of these to froth. The material, if in solution, is added to the digestion flask by means of a calibrated Ostwald pipette; if in solid form, as plant organs or sections, it is carefully dried and weighed. The quantity of material taken for digestion must be deter- mined by a preliminary rough analysis, since the method is best adapted for amounts of nitrogen between .5 and 5 mg. One сс. (more if needed) of chemically pure sulphuric acid (сопс.) is added to the material to be digested, the amount depending upon the quantity of carbon-containing compounds present, then 1 gram of potassium sulphate and a drop of 5 per cent copper sulphate. The contents are heated slowly until frothing is over, after which a hotter flame may be employed. Sometimes it is necessary to add some solid frag- ment to prevent bumping, a bit of unglazed porcelain being especially good. Small mica chimneys can be obtained to protect the flames from air currents, but lacking these bot- tomless beakers may be used. Upon completion of digestion the contents of the flask are permitted to cool somewhat (the liquid must not become 1 The condensers can be made in the laboratory from glass tubing. 1916] DAVIS—FOLIN MICRO-KJELDAHL APPARATUS 409 solid), then 50 ce. of ammonia-free water carefully added. Folin removed the nitrogen by adding saturated NaOH to alkalinity, then forcing the ammonia over into standard acid with a vigorous air current. While the method is excel- lent with the small amount of water used in the Jena tube, it does not give good results with the Kjeldahl flask and the larger amount of water used there. Distillation is more efficient. Small condenser tubes were made in the laboratory from glass tubing, the outer jacket measuring 402 сш., and Ше inner 5 mm. in diameter. The lower end of this latter, where it dipped into the collection acid, was fitted with a larger tube 14 mm. in diameter—this to prevent back-flow of the acid; to the upper end of the inner tube was attached a safety trap made from a 10-сс. pipette, which, in turn, fitted into the Kjeldahl flask by means of a two-hole rubber stopper. Through the second hole of this was inserted a small piece of glass tubing closed at the upper end with a bit of rubber tub- ing and a pinch clamp, thus making it possible to add the alkali after the apparatus had been connected up for distilla- tion. The distillation is carried on in the usual way. It is com- monly necessary to add a pinch of zine dust to the distilling mixture to prevent bumping, while a few drops of liquid par- affin will keep down a tendency to froth. The ammonia is col- lected in N/20 acid and titrated against alkali of the same strength. Alizarin red (Alizarin sulfonsáure Natrium, Merck) in .1 per cent aqueous solution gave best satisfaction as an indicator. Folin has chiefly employed the colorimeter for the actual determination of nitrogen. The method has its distinct ad- vantages, especially if the precautions indieated by Folin are observed in Nesslerizing. In the absence of a colorimeter, however, and because excellent results were always obtained in our work by titration, the latter method has been retained. The following tables show how the results obtained with the ‘‘micro’’-method approximate very closely those gotten with larger amounts of material in the original Kjeldahl. The ilustration is that of an ordinary laboratory experiment [Vor. 3, 1916] 410 ANNALS OF THE MISSOURI BOTANICAL GARDEN showing some of the steps involved in the enzymie hydrolysis of albumin. Papain (.1 gm.) was added to 200 ee 2.5 per cent albu- min solution, alkalinity reduced to N/250, then incubated at 40°C. for two hours. Portions were removed and tested both with the ‘‘micro’’- and the ‘‘macro’’-Kjeldahl for proteolytic change. Опе се. was digested with the former, 15 се. were used with the latter. NITROGEN DETERMINATIONS IN THE HYDROLYSIS OF ALBUMIN Micro-Kjeldahl | Macro-Kjeldahl Nitrogen Albumin | Albumin |Albumin |Albumin |Albumin |Albumin fractions +епғуте | +water |--епгуте| -+water |-+enzyme| +water in 1 cc. calc. to 15 cc. in 15 cc. mg. mg. mg. mg. mg. mg. Total nitrogen.... 3.80 3.795 57.00 56.925 57.615 57.500 oagulable nitrogen........ .913 3.145 13.695 | 47.175 14.125 48.225 Phosphotungstate РЧ р 1.397 .472 20.955 7.080 21.275 6.925 Ee acids and МА ree ree 1.490 177 22.35 2.655 22.5025 2.785 Further comparison is made in the recovery of nitrogen from a carefully prepared solution of (NH4)»SO,. As be- fore, 1 ec. of solution was used with the ‘‘micro’’- and 15 се. with the ‘‘macro’’-Kjeldahl. TABLE II THE RECOVERY OF MEE ter A SOLUTION OF (NH4:S50. CONTAINING 217 MGS. PER CC. Micro-Kjeldahl, Macro-Kjeldahl Exp. 1 cc. solution used 15 cc. solution used no. Found Calculated for 15 cc. Found mg. mg. mg. 1 3.13 46.95 47.595 2 3.12 46.80 48.125 3 3.17 47.55 46.925 4 3.185 47.775 45 .400 5 3.192 47.880 47.3925 Theoretical 3.217 48.255 48.255 Graduate Laboratory, Missouri Botanical Garden. [Vor. 3, 1916] ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 7 Folin’s micro-Kjeldahl apparatus for the determination of nitrogen. 4 PLATE 1916 w Yw 2% .— H. DAVIS—FOLIN MICRO-KJELDAHL APPARATUS STUDIES IN THE PHYSIOLOGY OF THE FUNGI?! I. NITROGEN FIXATION B. M. DUGGAR Physiologist to the Missouri Botanical Garden, in Charge of Graduate Laboratory Professor of Plant Physiology in the Henry Shaw School of Botany of Washington University AND A. R. DAVIS Formerly Research Assistant to the Missouri Botanical Garden INTRODUCTION AND CRITICAL REVIEW oF LITERATURE The problem of the fixation of free (atmospheric) or molecu- lar nitrogen by the fungi has received attention at the hands of no small number of investigators, yet a careful study of the literature is sufficient to indicate that much further work —with the strictest regards for accurate methods—will be re- quired before the problem is satisfactorily solved. For rea- sons developed later in this paper, we have felt the desir- ability of continuing, under different conditions, the investi- gations begun by one of us some years ago. At the present time there can be no doubt entertained, of course, as to the capacity of the legume tubercle bacteria (Bacillus radicicola vars.) and certain soil forms (notably Azotobacter spp. and Clostridium Pasteurianum) to fix nitro- gen. Here the amounts of nitrogen-increase in relatively small cultures under favorable conditions are so far above any regu- lar experimental errors, and so consistently reported by care- ful workers, that the simple question of whether or not there is fixation is eliminated. On the other hand, there is much con- tradictory evidence as to the fact of nitrogen fixation by other bacteria and by the fungi, especially by the moulds and 1 NorE.—About half a dozen investigations are already in progress dealing with the physiology of the fungi, and it is proposed to give considerable attention to this phase of physiology during the next few years. The investigations would include certain aspects of nutrition and enzyme action, growth relations—espe- cially the effects of environmental factors—and various phases of the general “Studies in the Physiology of the Fungi," of which the present article is No. 1. --В. M. DvGGAR. ANN. Мо. Bor. GARD., VoL. 3, 1916 (413) [Vor. 3 414 ANNALS OF THE MISSOURI BOTANICAL GARDEN other saprophytie species. Even those who report fixation for the last-mentioned fungi base their conclusions, in the majority of cases, upon amounts which are only questionably beyond the possibility of experimental error. The literature has been frequently reviewed, but for purposes of discussion in this and in forthcoming papers, it has been found well to give this detailed consideration. No account of N-fixation by bacteria is included. The experiments of Jodin (’62) are now interesting merely in an historical way. He observed fungi, in an impure culture, to grow upon media, ‘‘trés sensiblement exemptes de com- posés azotés organique ou mineraux." Employing the methods of gas analysis, he found that in a sealed vessel the amount of molecular nitrogen used was from 6 to 7 per cent of the oxygen consumed. Hallier (’67) simply reported that he had often observed yeasts growing in a nitrogen-free medium, but he realized the necessity of quantitative data in order definitely to deter- mine the fact of nitrogen fixation. From observations on the growth of mould fungi in nitrogen-containing and nitrogen- free media Nägeli (780) concluded that mould fungi are unable to assimilate free nitrogen. Frank ('92) states that he grew certain forms of Peni- cillium upon nutrient media lacking nitrogen and then tested the media for the presence of nitrogen, securing a positive indieation. No quantitative work appears to have been done, nor are the quantities of medium employed in either case men- tioned. Later he (793) reported growing Penicillium clado- sporioides! (—Hormodendron cladosporioides) during ten months in nitrogen-free media containing sugar. A culture on 65 ce. of solution is reported to have given a fixation of 3.5 mg. nitrogen. Insufficient details are furnished regard- ing methods employed and the use of controls. Using Aspergillus niger and Alternaria tenuis as the basis of a test, supplementary to a more complete study of bacteria, Berthelot (793) reported a fixation of 5.8 to 10.0 mg. in the 1 In the discussion of literature the writers have written mg names of the various organisms just as they are given in the original article 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 415 first-mentioned fungus, and 4.6 to 11.1 mg. in the last-named species. This appears to be the total amount fixed after growing for a period of months in 600 се. of Cohn’s solution with various sources of carbon. For 100 се. the nitrogen quan- tities would therefore be .97-1.67 and .77-1.85 mg., respec- tively. Fixation by Gymnoascus is also mentioned. With respect to analytical methods the exact procedure is not given, and one might, perhaps, without being too critical, wish to have had assurances regarding the purity of cultures, the nature of the vessels (‘‘ballons’’) used, and how the sample for analysis was taken, especially in view of the following remark made in regard to the amount of fixation in one of the series with bacteria: ‘‘Ils auraient été sans doute plus accusés si la dessiecation des matériaux n'avait pas fini par amener la mort des bactéries." Puriewitsch (795) used Aspergillus niger and Penicillium glaucum in a nutrient salt solution containing also 3 per cent tartarie acid, variable amounts of cane sugar, also small amounts of ammonium nitrate. He obtained a mean nitrogen fixation of 4.51 mg. for Aspergillus, and 3.26 mg. for Penicil- lium. It is not stated whether these amounts are calculated on the basis of 100 ce. of solution, or whether they were for 25-50 ce., the quantities which appear to have been used in the different cultures. At the same time he reported that the amount of fixation inereased with the concentration of sugar, but did not increase in direct proportion to the increase in weight of the mycelium. It is not clear, though quite possible, that these results were obtained with pure cultures. More- over, since in some of the cultures, at least, dry weight de- terminations of the fungous felt were made, the conclusion is unavoidable that herein might be a possibility of error. Like- wise, the division of some of the experiments into ‘‘(a)’’ and “(b)” suggests that the whole of the solution was not em- ployed in the analysis. The results were subsequently eriti- cized by Czapek (701) and Heinze (706) on other grounds. The fact that fungi may make an appreciable growth on media eontaining a very low minimum of nitrogen, without nitrogen gain, has been pointed out by some investigators, [Vor. 8 416 ANNALS OF THE MISSOURI BOTANICAL GARDEN and may adequately explain Fermi’s (’96) statement to the effect that he was able to grow certain moulds and yeasts on nitrogen-free media without nitrogen fixation. Brefeld (’00) determined that cereals and grasses infected with species of Ustilago were unable to assimilate free nitro- gen, but this type of negative evidence is valueless in the present discussion. More extensive than any of the earlier work is that reported by Saida (’01) who investigated seven species, three of which (Phoma Betae, Mucor stolonifer, and Aspergillus niger) give nitrogen fixation both with and without the presence of com- bined nitrogen in the culture medium, one species (Endococ- cus purpurascens) requires the presence of combined nitro- gen, and three species give negative results. In most cases the fungi were grown on 50 ce. of nutrient salt solutions con- taining dextrose or cane sugar, the source of nitrogen being a small quantity of (NH4)2SO4 or of (КН.)„СОз. In none of the four species except Phoma Betae is the amount of fixa- tion more than about 2 mg. (.8871-2.0699 mg.). Fixation (varying from 1.1828 to 10.536 mg.) in P. Betae rises some- what in relation to sugar content of the medium, although maximum fixation occurs in sugar beet decoction plus sugar. The exact method of handling the cultures is not described, but with the exception of possibilities mentioned later, the work seems to be above criticism. Czapek (201) states that Aspergillus niger does not fix free nitrogen. Later, as a result of numerous experiments on nitrogen-containing media Czapek (’02) again reports no fixation for this species. He declares that the work of Purie- witsch and Saida requires confirmation. Studying the effects of a yeast and a mould on the nitro- gen fixation of Azotobacter, Gerlach and Vogel ('03) con- clude from analyses of the control cultures, in which each of these organisms was grown alone, that neither of the former fungi are capable of utilizing atmospheric nitrogen. Koch (’03) was unable to demonstrate any nitrogen fixa- tion for Aspergillus niger in a few preliminary experiments. He draws attention, however, to an experiment made by 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 417 Hiltner from which it would appear that Lolium temulentum (inhabited by an associated fungus) thrives equally well in quartz sand with or without nitrogen as fertilizer, while the fungus-free Loliwm italicum develops much better when the quartz sand is fertilized with nitrogen than when the sub- stratum is without such fertilization. In the first experiments reported by Ternetz (’04) with the fungus isolated from the roots of certain Ericaceae, and later designated Phoma radicis vars., very slight nitrogen fixation was found. In 100-150 ce. of nutrient solutions containing dextrose .6-3.85 mg. represent the range of fixation. Stimulated by the work of Saida and others, Heinze (’06) reports a detailed repetition of the work of Saida (’01) and Puriewitsch (795), employing to a considerable extent the same organisms and the same solutions. The work seems to have been unusually extensive, but since it was in every case negative, no details are published. Heinze was apparently inclined in 1903 to consider the possibility that yeasts in cer- tain stages may fix nitrogen, since he states (see review of Schulze’s work by Heinze, Centralbl. f. Bakt. IL. 10: p. 675): ‘‘Schliesslich deuten mancherlei Beobachtungen der Ref. darauf hin, dass man auch event. bei den Hefen—und zwar in statu sporulandi—moglicherweise gerade bei den Vorgängen, bei denen die Spore nach Hansen wiederum zum Sporangium wird, mit gewissen mehr oder weniger stark ausgeprügten N-Assimilationsvorgángen zu rechnen hat." This earlier statement is apparently the basis of Lipman's (’11—’12, see p. 173) reference to Heinze's work. Through pot experiments with seedlings of Pinus montana with and without mycorhiza, Moller (206) concluded that the fungus associated with the roots of this species is unable to supply the host with nitrogen accruing as a result of fixation. In continuation of her earlier work Ternetz (’07) has secured data of special interest, with reference to fixation, for Phoma radicis vars., likewise in a comparative way for a few other fungi and bacteria. The utmost care seems to have been observed with respect to the purity of materials, the use of necessary blanks in the analyses, and of controls in the [VoL. 3 418 ANNALS OF THE MISSOURI BOTANICAL GARDEN experiments. Cultivated during a period of 28 days on 50 ce. of nutrient solution the 5 races of Phoma radicis gave a nitro- gen fixation ranging from 2.3 mg. in the lowest to 15.7 mg. in the highest. For Aspergillus niger and Penicillium glaucum the fixation was 1.9 and 2.8 mg., respectively. The usual high fixation was secured with Azotobacter chroococcum and Clos- tridium pastorianum. In spite of the fact that the methods employed are those generally recognized as unimpeachable, still attention should be drawn to the fact that in such cases as those of Aspergillus and Penicillium, where the fixation is only about 2 mg., the technique employed must be subjected to the closest scrutiny. It is noted that the felt was separated from the culture solution in the usual way, and further that the solution was then made up to the original volume (a pro- cedure of vital importance when aliquot parts of the solution serve for analysis, and one seldom mentioned). Then from one-sixteenth to one-fourth, depending upon the amount of sugar present, of the total solution was taken for the analysis. In this way any small experimental error involved would have been multiplied 4-16 times. Equally satisfactory in respect to method is the work of Froehlich (’08). Here again the methods are described in sufficient detail so that one is not left in doubt as to import- ant parts of the technique. The organisms used lend a par- ticular interest to the work, inasmuch as they were isolated from dead and decaying plant material and are fungi gener- ally considered important in the decay of vegetation. Those selected consist of one species from each of four common genera. All were found to fix nitrogen to a slight degree, averaging as follows: Alternaria 3.34, Macrosporium 3.70, Cladosporium 2.26, and Hormodendron 1.93 mg. Many sub- sidiary experiments of interest are included. Zikes (209) conducted extensive experiments to determine the free nitrogen relations of a yeast-like organism isolated from the leaves of laurel and called by him Torula Wiesneri, which he cultivated on flasks containing 300 ce. of culture fluid. He employed the Dumas method of analysis, filtered off the fluid from the yeast cells, and made separate determina- 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 419 tions. He reports a fixation of 5.1-6.5 mg. per liter, .51-.61 per 100 ec. of culture solution, which, however, he regards as satisfactory positive evidence. No investigator has obtained figures comparable to those of Latham (709). This work was well conceived and ar- ranged with a view to determining the effect of zinc sulphate on the nitrogen fixation of Sterigmatocystis migra (Asper- gillus niger) abundantly supplied with combined nitrogen.! The results published exhibit a variation ranging from a nitrogen loss of 42.5 mg. to a fixation of 205.1 mg. per culture, on 90 ce. of medium. In view of all the earlier and later studies made on fixation by this fungus, granting at the same time, of course, possible differences in strains, it can only be surmised, perhaps, that miscalculations are accountable for these unusual results. It would appear that in making the analyses she employed aliquot parts of the culture solution, and likewise divided the felt. Such a procedure, however, only suggests possibilities and cannot explain the results. In the case of maximum fixation, 677.3 mg. of nitrogen are re- ported fixed in felt and solution per gram of dry felt pro- duced. This is an amount incomparably greater than any- thing elsewhere obtained. Duggar and Knudson (711) reported only by abstract upon extensive series of experiments in which Aspergillus niger, Trichoderma lignorum (erroneously given as T. lignicola), and several species of Basidiomycetes were employed. Var- ious nutrient media were used, including synthetic nutrient solutions, leaf decoctions, and decayed leaves ground to a fine powder. None of the cultures showed a difference in the N-content over the controls sufficient to indicate fixation, whether with or without combined nitrogen. It may be stated that this work was not published in detail by reason of the uniformly negative results. It was intended to pursue the work further using ground leaf mould and similar materials as nutrient media, but difficulties in obtaining uniform samples The nutrient salt solution employed was the well-known Richards’ solution patton ^ ХН,ХОз 1 gm., КНоРО, 0.5 gm., MgSO, 0.25 gm., FeCls trace, and sugar 5 gm., except that the amount of the nitrate in the different series varied from 115.4 to 160.3 mg. per culture, or 50 cc. of solution. [VoL. 3 420 ANNALS OF THE MISSOURI BOTANICAL GARDEN for analysis on such media were almost insuperable. The de- sirability of growing the Basidiomycetes on solid media was obvious, but under such circumstances it would have been necessary for greatest accuracy either to analyze the entire contents of such bulky cultures or to take account of change in weight of materials due to loss of СО» and H20 produced in respiration. Lohnis (210) seems to report having found N-fixation in Torula, but his work was neither extensive nor reported in such way as to give the details of the methods employed. Pennington ('11) worked with two species of Penicillium, two species of Fusarium, Aspergillus niger, and Alternaria sp. A variety of experiments was arranged in liter flasks containing 100 ec. of solution. Employing aecepted methods he obtained no nitrogen fixation in a first series of experi- ments, although there was some growth on media practically without nitrogen, that is, with scarcely perceptible amounts, due, he believes, to impurities in the dextrose employed. In another extensive series no differences between the flasks con- taining the moulds and the controls were sufficient to indicate fixation. In one ease Penicillium gave an apparent fixation of .88 mg. In considerable part his work was planned to con- firm or disprove that of Latham (209). The results are therefore peculiarly interesting, especially as he has ap- parently observed great care to eliminate all possibilities of error, and in advance thoroughly tested his ability to deter- mine the nitrogen content of the eultures accurately. Medisch (710) observed some growth of the fungus Hypo- crea rufa in solutions to which no nitrogen was added. He reports a gain of 1.05-2.45 mg. nitrogen in 50 ce. distilled water. This preliminary experiment was followed by others in which, under purified air, the organism was grown on var- ious culture solutions. "These included solutions containing no nitrogen, nitrogen as ‘‘potassium humate,’’ and as NH4NO3. The results indicate that, whereas in the first case, with nitro- gen present only as an impurity, the fixation was 1.74—3.23 mg. in 100 cc., in the humate solution the gain was 3.15-4.61 mg., and in the solution containing NH4NOs Ше gain was 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 421 2.45-3.06 mg. He considers these quantities as possibly within the limit о” experimental error. Unfortunately, various de- tails regarding the handling of cultures and the methods of analysis are omitted. Lipman (711-7112) made an extended study of the relation of certain yeasts and fungi to nitrogen fixation, employing 7 species of yeast, 5 pseudo yeasts, and Mycoderma, also Aspergillus niger, Penicillium glaucum, and Botrytis cinerea. In the first extensive series, omitting the moulds, there was a slight gain of nitrogen in practically all cases, but only in a single case, a pseudo yeast, was this more than 1.0 mg. In a second series 9 out of 15 forms gave a gain of 1.05-2.28 mg., while 6 forms yielded less than 1 mg. increase of nitrogen. In the final series, which includes the fungi, the yeasts exhibited gains of .07-3.78 mg., while the fungi ranged from .05 to 2.38 mg. Preliminary to her studies in nitrogen fixation Stahel (211) made an extensive trial of fungi on media low in nitrogen. She isolated 54 species, largely from decaying vegetation, and then grew these on various media, including silica jelly with- out combined nitrogen. Five species were found to grow well on the last-named medium, while 22 made some growth. In the studies regarding N-fixation it is determined that the 5 species which grew on nitrogen-free media are capable of fixa- tion, likewise 4 of the organisms from the group growing in- differently also possess this capacity. The method of hand- ling the flasks is not given in detail, but it appears that the mycelium was filtered off from the solution and separate analyses made. The fungi were grown on 200 ce. of nutrient solution, and, in general, the amount of fixation was related to the initial nitrogen content. While the method suggests some possibilities for error, yet in some cases the amount of fixation is certainly well above the usual experimental errors. It is to be noted, however, that the following amounts repre- sent questionable fixation: Aspergillus .41 mg., Penicillium .50 mg., Botrytis 46 mg., Melanoma Ap mg., Epicoccum .41 mg., Bispora .61-1.44 mg.; while the following give higher re- [Vor. 3 422 ANNALS OF THE MISSOURI BOTANICAL GARDEN sults: Alternaria 1.02-5.55 mg., Hormodendron .36-5.0 mg., Macrosporium .23-5.91 mg. Experiments conducted by Kossowiez ('12) which seemed to suggest N-fixation in the case of certain species of Sac- charomyces, Monilia candida, and Oidium lactis were subse- quently repeated by him (714) under more nearly standard control conditions. The results were interpreted as entirely negative. Besides the organisms previously employed, he used also Aspergillus niger, A. glaucus, Penicillium glaucum, P. brevicaule, and a species of each of the following, Botrytis, Mucor, and Isaria. Will (212), reporting work of Scheckenbach, declared the capacity of certain species of Torula to grow upon nutrient media lacking nitrogen, likewise to fix atmospherie nitrogen when little or no combined nitrogen was supplied. "There is, however, with the experiments reported, little evidence that sufficient precautions were taken in the arrangement of suit- able controls. The capacity of Blastoderma salmimcolor, Torula sp., and **pastorianus"" yeast to fix nitrogen has been mentioned by Lindner (712),! but to what extent this work was quantita- tively executed cannot be determined from the data at hand. Goddard's (713) investigations parallel those of Froehlich, Stahel, and, to a certain extent, those of Ternetz. He isolated 15 species of fungi from the soil, and tested each of these with respect to nitrogen fixation, grown on 50 cc. of a culture solu- tion eomparable to the nutrient media employed by other in- vestigators. Every possible precaution seems to have been taken to insure accuracy. The fungi were grown 48-70 days. With no organism in any series were there indications of con- sistent gains over the initial nitrogen content. Species of Aspergillus and Penicillium were included in these studies. In connection with his investigations of mycorhiza problems, Peklo (713) isolated 3 species of fungi, 2 being species of Penicillium, and one an indetermined form. Each of these was grown on Winogradski's solution plus dextrose for 1-2 1 The reference given appears to be an abstract of a more extensive report which is at present unavailable. 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 423 months. For each species he claims positive results, the fixa- tion ranging from 0.8575 mg. in the lowest to 1.8615 mg. in the highest, per 100 ec. of solution. The inference seems to be that in each case a single inoculated culture or a single con- trol was usually employed. It is of interest to note that aside from the few analyses made, the fungous felt and the solution were separately analyzed. Traaen (714) made no quantitative studies to determine N- fixation, but he observed the growth of 4 fungi on media prac- tieally nitrogen-free, and as a result of the very weak growth he came to the conclusion that under the conditions they could not possibly utilize atmospheric nitrogen. Using strains of Aspergillus niger and Penicillium glaucum, Chambers (716) was unable to demonstrate any N-fixation. He employed Folin’s micro-Kjeldahl method, growing the organisms in long Jena test-tubes and making the determina- tions without transfer of any portion of the culture. METHODS The organisms used in this work were Aspergillus niger, a strain long employed in various physiological experiments in this laboratory; a species of Penicillium, isolated from leaves and corresponding closely to Thom's idea of P. expansum; P. digitatum, isolated from a decaying orange; Macrosporium commune, isolated from dried grass culms; Phoma Betae, a culture obtained through the kindness of Mr. E. C. Rittue, Los Angeles, California; and for comparison three forms of Azoto- bacter, as follows, all three being furnished by Dr. J. G. Lip- man, A. vinelandii, A. chroococcum (from Kansas soil), and A. chroococcum (from Colorado soil). Except as to the source of nitrogen and carbon, there has been no great dissimilarity in the mineral nutrient solutions employed by European investigators. The Cohn solution or a modification of it has been the basis of much of the foreign work. We wished to have some of our experiments follow fairly closely the work of Saida, therefore we have used in 1 The NE and cultural characters of this organism will be described in a subsequent paper [Vor. 3 424 ANNALS OF THE MISSOURI BOTANICAL GARDEN some of the experiments (series 1A-4A, table r) his solution as regards concentration of mineral nutrients. It is as follows: КН.РО, 4 gram MgSO š ESL CaCl à - See This is designated solution A. H30 100 се. To this has been added (NH4)2804 or asparagin as a source of nitrogen, and dextrose or saecharose as a source of carbon. In most of the work, however (series 5B-15B, 17B-22B, table т), it has seemed well to use a modification of the formula known as Richards”? solution, used especially by Miss Latham in securing the extraordinary results to be referred to later. The modification consists merely in varying the sources and amounts of nitrogen and carbon furnished, these last being the same as employed with ‘‘solution A’’ above. The Richards’ solution consisted of: КН.РО. 5 gram Men, 25 gram Ki I | FeCl; E This is designated solution B. H30 100 ee. Stock solutions of each constituent were made up of appro- priate strength, usually such that an equal quantity of each was required for any culture. For Glomerella Gossypii a modification of the Uschinsky solution, as indieated in table r, was employed, since this had been found satisfaetory for this organism through other workers in the laboratory. For the various strains of Azoto- bacter a soil-compost extract containing mannite was em- ployed. Three hundred gm. of potting soil and 100 gm. of well- fermented compost were each extraeted for 2 hours with 1 liter of water, then filtered, and the filtrates combined. To the mixed extract was added for each 100 ee, the following con- stituents: K»HPO, .05 gm., CaCO; 1 gm., and mannite 5.0 gm. Kjeldahl flasks of 500 cc. capacity were used as culture ves- sels in all cases, and into each were placed 50 ce. of the solution required.! The idea of using the Kjeldahl flasks for the cul- 1 In the bacterial cultures 100 cc. of solution were employed. 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 425 tures enabled us to make the nitrogen determinations of both inoculated and uninoculated flasks from the entire contents of the flasks, therefore to dispense entirely with any transfers of culture solution or fungous felt, and to avoid the possibility of errors thus ensuing. All glassware was cleaned by standard methods; nitrogen- free double distilled water was used; and Merck’s reagents. Every experiment was set up in triplicate, also with three controls; that is, for every series in which a different fungus, a different amount or source of nitrogen or of carbon was used, there were 6 cultures, 3 of which were inoculated and incubated, while the remaining 3 were inoculated, autoclaved to kill the spores (since they served as controls), and were then incubated with the others. The inoculations were made from cultures on potato agar, fresh cultures only being employed as a source of spores or mycelium. The inoculation procedure was as follows for those forms producing spores: Numerous spores were transferred to a flask containing 100 ec. sterile НәО. This was agitated until there was an evident spore suspension, and this then pipetted out with a sterile pipette into a second sterile flask. From this last flask Lee, portions were transferred with sterile pipettes to each flask in the series. The controls were then autoclaved for 15 minutes at 15 pounds pressure. That the method was entirely satisfactory is shown by the fact that there was only a single case of contamination in all the series employed and no case of growth in any of the controls. Sim- ilarly, in the inoculation of the series with Azotobacter, loops of the organism were diffused in sterile water, then Le, portions were placed in each flask by means of a sterile grad- uated pipette. All transfers were made with the greatest pre- caution in a steamed transfer room. In the case of Phoma Betae, where no spores were produced, small masses of hyphae of approximately equal size were inserted into each flask. Repeated tests have shown that in the incubator rooms for the length of time which these experiments were permitted to run there is no detectable amount of combined nitrogen ab- sorbed by flasks of the culture solution or by flasks of dis- [Vor. 3 426 ANNALS OF THE MISSOURI BOTANICAL GARDEN tilled water. Both from this fact and further from the nature of the controls it was unnecessary to place the cultures in a chamber arranged to protect against combined nitrogen. The data presented in this paper on the determination of nitrogen were obtained either with the Kjeldahl-Gunning method,—using mercury in addition to potassium sulphate,— or, where nitrates were involved, with the Forster modifica- tion of the method mentioned. In some preliminary work an extended attempt was made to utilize the Folin micro-Kjel- dahl apparatus, but that proved inapplieable to the present work for the following reason: The amount of culture solution which it is possible to use with this method is small, and doubt- less would be too small to yield convincing results in view of the present confusion regarding the question of nitrogen fixa- tion in the fungi. In the light of the results obtained by Purie- witsch, Saida, and others, all of whom used from 50 to 100 сс. of culture solution, it seemed essential to employ the ‘‘macro’’ method and to deal with cultures as large as practicable. In the pioneer work of Jodin ('62) gas analysis methods were employed for the determination of nitrogen fixation, therefore through the indirect method of nitrogen loss in the culture chamber. Since that time all the work which may claim a right to be considered quantitative has been made with the Kjeldahl method, or with some modification of it, usually the Gunning. That this method is sufficiently accurate to detect any amount of fixation worthy of the name is evident, since an experienced analyst can usually secure results which often check to within .2 mg. However, if one does not observe all possible precautions, errors may creep in which will yield widely varying results. Chief among these possi- bilities in the problem of nitrogen fixation are the following: Impure chemicals. . 2. Accuracy of standard acid and alkali. 3. Indicator. 4. Completeness of digestion and distillation. 5. Loss of nitrogen in the transfer of the culture material, or felt, from one flask to another. x 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 427 6. Multiplication of the experimental error through taking an aliquot part of the fungous mat or culture solution and upon the determination from this basing a calculation for the whole. 7. Inadequate controls. Analyzed chemicals may be obtained always, but these should be checked by running blank experiments. Standard acids and alkalis should be checked up by at least two methods. Nevertheless, slight discrepancy in the standard affects the actual rather than the relative analytical results, provided the same solutions are used for the nitrogen determinations whether they grow the fungus or are used as controls. Certain indicators have, in the presence of ammonia, what might be called a ‘‘running’’ end-point; that is, the color change occurs through a fairly wide range of H-ion concentration. After trying several indicators for this work alizarin red (Alizarin sulfonsáure Natrium, Merck) and cochineal were found to give the best satisfaction. The former in .1 per cent aqueous solu- tion was used. The error due to incomplete digestion or distillation, while easily guarded against, may sometimes occur, if care is not observed. It was the practice here to continue the digestion 15 minutes after the mixture had become colorless. А full hour was given to distillation, since this interval proved en- tirely sufficient as shown by tests from time to time. In the critical review of literature it has been emphasized that many of those investigators reporting nitrogen fixation forthe fungi have limited their nitrogen determinations to aliquot portions of the culture solution. The total nitrogen was then calculated. Summarizing some of the points to which attention should be drawn, it is found that Puriewitsch (295), Saida (201), Ternetz (’07), and Froehlich (208) all filtered the solution from the fungous mat, determined the nitrogen from a portion of the solution, and calculated for the whole solution. The mat nitrogen was determined separately. Stahel (211), Peklo (713), and others, after separating solu- tion from fungous felt, evaporated the culture medium to small bulk (following the addition of acid) and determined the [VoL. 3 428 ANNALS OF THE MISSOURI BOTANICAL GARDEN total nitrogen. To this was added the amount of nitrogen found in the felt. Lipman (711-7112) did not separate the mat from the medium, but transferred the whole to a digestion flask, and later to a distilling flask, determining the total nitro- gen in one lot. All the cases cited above involved one or more transfers of material, since the fungi were usually grown in Erlenmeyer flasks or similar receptacles, and the contents filtered or transferred before digestion, thence usually a sec- ond transfer to a distilling flask. It was with the end in view of eliminating the possibility of error in this direction that the method already described was employed, i. e., of growing, digesting, and distilling in the same flask and without transfer. Where the digestion of nitrates was involved in the culture solution, the previous investigators have used, almost with- out exception, the Gunning-Jodlbauer method—phenol or salicylic acid and zine dust being employed for the reduc- tion of nitrates. In our work the Forster modification was employed, since certain workers have found difficulty in ob- taining all the nitrate by the former method. Indeed, it was this difficulty which first led Förster to use sodium thio- sulphate as a reducing agent. If all nitrates are not reduced a serious error is, of course, involved, one which, moreover, makes for a difference between controls and inoculated flasks. The results may be presumably correct for the converted or assimilated nitrogen of the mycelium (or products excreted therefrom), low figures resulting for the nitrate of the cul- ture media. If an error were present, then, it would be related somewhat closely to total growth or to sugar consump- tion, factors determining nitrate consumption. It is equally true that the capacity to fix nitrogen by a fungus, if possible, might also be related to the capacity for growth under the particular conditions. In the use of the Forster method at first certain difficulties were experienced. In preliminary work the recovery of nitrogen from a water solution of KNOs was easily accom- plished within experimental error. When, however, a nitrate was added to a soil, compost, or plant tissue decoction the results were invariably low. It was found necessary to add 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 429 more sodium thiosulphate (3 gm. instead of 2) and to allow 10-15 minutes after its apparent decomposition had taken place before digestion was continued. This illustrates the possibility of error in a method that is not thoroughly tested in connection with the peculiar conditions at hand. No difficulty was experienced in obtaining results with the Gunning-Kjeldahl modification that checked within experi- mental error. Some trouble, through frothing, will be ex- perienced in the actual digestion, however, where the culture media are high in sugars. This may be overcome by boiling (after adding 15 ee. of concentrated sulphuric acid) slowly for an hour or more, then adding more acid together with 15 gm. of potassium sulphate and 1 gm. of mercury. In our work it became necessary at times to add a third lot of 15 се. acid—the same amount being always added to both fungus-containing flasks and controls. Distillation was carried out through block tin tubes which had been in use sufficiently long to obviate the possibility of error through absorption of ammonia—a point observed with new tin by several investigators. The standard acid and alkali were restandardized at short intervals. The same lot of chemicals was always used throughout a single series to insure parallel treatment with both fungus-containing flasks and controls. EXPERIMENTAL RESULTS AND DISCUSSION The results of our experiments are presented in some de- tail in table т. It is necessary to note that while the quantities given in column v were obtained by careful weighings, they represent only approximately the quantities present in the solution as determined by analysis (see columns vi and vir). In any series the controls are as nearly perfect as we were able to arrange, that is, the solutions in all the flasks in any one series were taken from a single vessel of the culture medium, the complete mixing of the different constituents in the culture solution being given special attention. In column 1 the letters A and B given in connection with the series num- bers refer to the two nutrient salt solutions employed as de- 430 [Vor. 3 ANNALS OF THE MISSOURI BOTANICAL GARDEN scribed on page 424. Where no letters are given there are sufficient indications in column v to identify the culture medium employed. Data for all of the flasks analyzed are included in the table in order that the extent of the experi- mental error may appear just as well as the average of the determinations made. TABLE I NITROGEN FIXATION IN CERTAIN FUNGI AND BACTERIA I II III | IV V VI VII VHI- otal N in Total Nin |р. а + Е Geer EN pe 224 там: a flasks, N-fixa- ый Organism °С, gr. and C supplied, gb mg tion : days per cent Comp. | Aver- | Comp. | Aver- | | data age data age 8. 1A |Aspergillus | 30 | 30| .7 asparagin | 62.51 2.510 парег..... 3.6 dextrose | 62.545 |62.732| 62.335 | 62.382] .350 63.14 3 2А |A.niger.....| 30| 30| .7 asparagin | 61.215 1.915 1.8 dextrose | 61.985 | 61,915 | 59.710 | 61.180} .735 2.545 1.915 ЗА |А. niger....| 30 | 30| .014 asparagin | 2.135 2.310 108 dextrose | 1.925 | 1.960] 1.995 | 2.018 | —.058 1.820 1.750 4A |А. niger..... 30 | 30| .7 asparagin | 60.30 60.375 18.2 dextrose | 59.955 | 60.083 | 60.585 | 60.488 | —.405 59.990 SB |А. niger.....| 30| 30| .014 (КН) ХО 1.435 1.470 3.6 extrose 1.575 | 1.470| 1.505 | 1.470 | ———— 1.400 1.435 6B A. niger ...| 30 | 30 i (NH.):SO, 70.38 0.49 18.2 dextrose 70.420 | 70.417 | 70.490 | 70.547 | —.130 70.455 0.560 7B |А. niger.....| 30| 30| .014 (КН) ХО 1.71 .68 18.2 dextrose 1.715 | 1.715| 1.750 | 1.715 | ——— 1.715 8B |Macrospo- | 25 | 30] .014 (NH,):SO.| 2.030 1.925 rium сот- 18.2 dextrose 2.153 | 2.091| 2.065 | 1.995] .097 типе. .... 1.995 9B |М. commune 30| 7| 7 (МН,).50,| 70.38 0.42 18.2 dextrose | 70.420 | 70.373 | 70.560 | 70.548 | —.175 0.315 0.665 10В |M. commune| 30| 7 .014 a 1.755 .065 18. dextr 1.890 | 1.810| 2.205 | 2.065| —.255 1.785 .925 11B |M. commune| 25 | 30 tego bie ‚613 858 18.2 dextrose 753 dia -683 .747 | —.035 110 „100 1916] DUGGAR AND DAVIS—NITROGEN FIXATION TABLE I (Continued) NITROGEN FIXATION IN CERTAIN FUNGI AND BACTERIA 431 I II ІП | IV V VI VII VIII otal N in, Diff. - wv f Е CS Sources of N flasks containing cos p* N-fixa- да Organism °C. gr and C supplied, tion gf ан per cent Comp. | Aver- | Comp. | Aver- days mg data age data age 3 12B | Penicillium | 30| 42|] 7 (NHSO,| 70.455 70.630 digitatum . 18.2 dextrose | 70.630 | 70.513 | 70.560 | 70.618 | —.105 0.45 70.665 13B ІР. digitatum, 30 | 42 .014 (МН,):50,| 2.555 2.660 18.2 dextrose 2.590 | 2.602 | 2.765 | 2.730 | —.128 2.66 2.765 14B | Penicillium | 25 | 35 .7 (МН,),50,| 71.890 71.540 ехрапѕит 18.2 dextrose 72.065 | 71.978 ee 71.633 | .345 15В |Р. expansum| 25 | 35 .014 (NH)&SO,| 2.415 2.415 18.2 dextrose 315 2.403| 2.450 | 2.433 | —.030 16 |Glomerella | 25 | 30 | Uschinsky sol. 7.665 7.63 Gossypit.. cornmeal decoct. | 7.455 | 7.630} 7.875 | 7.758 | —.128 7.770 7.770 17B |Phoma Betae| 25*| 25 | mangel decoct. | 26.63 23.765 10.0 sucrose zi 26.997 | 23.695 | 23.730] 3.267 .98 18B |P. Betae....|25*| 25 ши decoct.| 16.975 14.665 10.0 crose ee 17.652 | 14.595 | 14.630 | 3.022 19.1 19B |P. Betae....|25*| 25 7 (NH>:SO,| 70.840 8.845 18.2 dextrose 70.735 |70.700| 69.020 | 68.927 | 1.773 0.525 8.915 20B |P. Веме.....25“ 25| .014 (NH;&SO.,| 2.275 2.030 8.2 dextrose 2.030 | 2.153 > 2.182 | --.029 21B ІР. Betae....|25*| 89 | mangel decoct. 53.340 45.220 10.0 sucrose за 52.990 | 45.255 | 45.238| 7.752 22B |P. Betae....|25*| 89 | sugar beet decoct.| 31.010 25.585 10.0 sucrose 31.360 | 31.185 | 25.655 | 25.620| 5.565 231 |Asotobacter | 25 | 28 — sol. | 46.51 5.810 in elandii 5.0 nnite eer 46.480| 6.405 | 6.108 | 40.372 244 |А. chroococ- | 25 | 28 аши sol, | 24.57 5.810 жик mannite 22.085 | 23.007} 6.405 | 6.108 | 16.899 (Colorado) 2.365 251 |А. chroococ- | 25 | 28 | soil-compost sol. | 22.586 5.810 cum.. 5.0 mannite 21.735 | 23.675| 6.405 | 6.108 | 17.567 (Kansas) 6.70 РУМИ од T In these cases only were 100 cc. of culture solution employed; in all other cases 50 cc. [Vor. 3 432 ANNALS OF THE MISSOURI BOTANICAL GARDEN From our results it is clear that under the conditions of the experiments no fixation can be claimed for Aspergillus niger, Macrosporium commune, Penicillium digitatum, P. expansum, and Glomerella Gossypu. For the most part, with these fungi, the differences between the various members of any series, including the controls, represent variations which might be expected, and the fact that the averages of the con- trols are slightly above or below those of the flasks contain- ing the fungi is of little significance. With Phoma Betae the case is different. Here the assimila- tion of free nitrogen seems definite. The quantities obtained vary from practically 0.0 to 7.75 mg. рег 50 се. of culture medium. All cultures on sugar beet and mangel decoction exhibit a nitrogen increase which points definitely to free nitrogen assimilation. It should be noted that these cultures represent series maintained for a shorter and a longer period of time; those maintained for the longer interval yielding higher fixation quantities than those cultured for the shorter interval. In one series, 19B, where the source of nitrogen is Л gram (NH4)2804, the nitrogen difference is perhaps suffi- cient to indicate nitrogen fixation. At any rate, if we regard fixation as occurring in this solution, it is fair to explain the absence of fixation in series 20B, in which only .014 (ХН.) 804 was employed, as due to the small amount of growth occurring in the last-mentioned series. As would be expected, fixation is somewhat related to the length of the period of growth and to the extent of growth. The results with Phoma Betae were so unexpected, in view of the long series of negative values obtained with other fungi, that a further check upon the work was introduced in the following way: A known amount of KNOs was added to a series of flasks containing 50 ce. of the sugar beet medium, and analyses were then made to ascertain with what accuracy this nitrogen could be determined. No difficulty was experienced in recov- ering this nitrate nitrogen, as shown by the data in table rr. Furthermore, it seemed well, as a result of the experiments with Phoma Betae, to employ by comparison certain organ- isms known to have nitrogen-fixing power. Accordingly, the 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 433 selected strains of Azotobacter were tested, and all yielded positive results of satisfactory magnitude, as shown in table 1. It will be observed in table 1 that slight discrepancies seem to occur between different series in respect to the amounts of nitrogen recovered—where the different series contained presumably the same amounts of initial nitrogen. This, how- ever, is only an apparent discrepancy, since, as previously TABLE II RECOVERY OF NITROGEN AS KNO# ADDED TO SUGAR BEET CULTURE MEDIUM Controls 13.8 mg. N : Trials (no N added) added as KNO; N d oo mg. mg. 8. в: 1 14.532 28.227 13.695 —.105 2 14.49 28.048 13.350 —.250 * 13.8 mg. nitrogen as KNO, added to 50 cc. sugar beet decoction + 10 per cent cane sugar. mentioned, this work extended over a considerable period of time, and although the same lot of culture solutions was used for the control flasks and for the flasks in which the organisms were grown in any one series, it was nevertheless necessary to make up new solutions from time to time for the different series. The different series are therefore only approximately comparable. Now since N-fixation occurs in organisms otherwise so physiologically different as Azotobacter, Clostridium, and Bacillus radicicola, why may it not occur in all fungi and bac- teria, it has been asked time and again. Final answer can be given only in accordance with the results of properly planned and carefully executed experiments. Moreover, it has been shown abundantly that fixation is relatively uncommon among bacteria, the capacity being possessed largely by those groups mentioned above. As has been indicated, among others, Saida, Ternetz, and Stahel have reported fixation for Aspergillus niger. Inthe cases referred to the results are scarcely greater than might occur as experimental errors. This could not apply, however, to the results with Phoma radicis and appar- ently not to those with Phoma Betae. Confirmatory evidence [VoL. 3 434 ANNALS OF THE MISSOURI BOTANICAL GARDEN from our own results has certainly designated the Phoma group of organisms as worthy of further careful study. With respect to the accumulated data for Penicillium, Macrosporium, Alternaria, and other saprophytic moulds oc- curring in the soil or upon decaying vegetation, it can only be said that the data fall into the same category as that for Aspergillus niger. We do not take issue with those reporting fixation, but we feel that in view of strong negative evidence regarding many of these fungi, further assurance must be given that the results may not be explained on the ground of experimental errors. We are quite well aware that the ad- mission of the data for Phoma Betae has virtually thrown open the whole question for any and all fungi, yet we can find no grounds upon which adequately to criticize either our own results or those of Ternetz with another species of this genus. Accepting the evidence for certain species of Phoma, in what direction shall we seek for organisms similarly endowed? Naturally related genera among the Sphaeropsidales would first be suggested, on purely morphological grounds. Again, for a long time physiologists have seen possibilities in organ- isms which have undergone such adjustment as characterizes the mycorhizal fungi generally. Up to the present time there has existed considerable uncertainty concerning the isolation and determination of the species which produce mycorhiza. Ternetz alone has demonstrated a Phoma as a root organism of this type. Peklo’s studies lead him to believe that Penicil- lium and an undetermined fungus are involved. In this case, as already noted, a very weak nitrogen fixation was reported. It is not intended, however, in this connection to discuss the various indications respecting mycorhizal fungi. Attention may be drawn to the fact that the predominant presence of Basidiomycetes in forests and meadows early suggested the association of these forms with the roots of higher plants. In recent times species of Tricholoma, Lactarius, Cortinarius, and Boletus have been strongly suspected of being important in the development of mycorhiza. The fungi are the primary agencies whereby vegetation is usually disintegrated or brought through the first stages of 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 435 decay. If it should be positively demonstrated, therefore, that the fungi concerned in this disintegration are at the same time capable of fixing an amount of nitrogen sufficient to prove of practical value, then it would be clear that agricul- tural practice might be modified in many ways to make greater use of this possibility of nitrogen enrichment accompanying the decay of herbage. As a matter of fact, however, the amount of fixation, as we have seen, reported for Alternaria, Macrosporium, Cladosporium, Aspergillus, Penicillium, ete., even by those recent investigators who claim fixation, is very slight—indeed, for such organisms it is usually considerably below 5 mg. per 50 сс. of solution. Assuming that there might be as much fungous felt in 1 cubic foot of ordinary soil as in 100 ce. of a culture! and that in both cases the amounts of fixa- tion might be equal, we would have as a maximum 10 mg. nitro- gen fixed per cubic foot or 420,000 mg. per acre, 1 foot deep, that is, 420 grams per acre, or about one pound. When it is recalled that in many cultures of Azotobacter the fixation has been as high as 50-200 mg. per 100 cc., and when it is further remembered that in the soil the conditions favor quantity of bacterial rather than fungous growth, we may perhaps gain some conception of the impracticability of claiming an eco- nomie relation in respect to nitrogen for such fungi. SUMMARY L A review is given of all available literature relating to nitrogen fixation by the fungi. 2. Culture and analytical methods are discussed, and sug- gestions are made with a view to the elimination of certain possible errors involved in this type of work. 9. Nitrogen fixation could not be demonstrated for Asper- gillus niger, Macrosporium commune, Penicillium digitatum, P. expansum, and Glomerella Соззури. 4. In eultures of Phoma Betae on mangel and on sugar 1 This seems highly EE in the light of recent discussion of this point; compare the following: Conn, H. J. Relative importance of fungi and bacteria in soil. Science N. 8. 44: 857-858. 1916. [VoL. 3 436 ANNALS OF THE MISSOURI BOTANICAL GARDEN beet decoction with sugar a nitrogen gain of 3.022-7.752 mg. was established, which seems definitely to indicate fixation. 5. Comparative studies of strains of Azotobacter exhibit the usual relatively large fixation of nitrogen in the culture media. Graduate Laboratory, Missouri Botanical Garden. BIBLIOGRAPHY Berthelot, M. (93). Recherches nouvelles sur les microrganismes fixateurs de P Compt. rend. Acad. Paris 116: 842-849. 93. Brefeld, O. ('00). Versuche über die Stickstoffaufnahme bei den Pflanzen. Schlesischen Ges. f. vaterl. Cultur. Zool.-bot. Sect., Jahresber. 78: 27—38. 1900. Chambers, C. О. (16). The fixation of free nitrogen by certain fungi. Plant World 19; 175-19. f. 1. 1916. Czapek, F. (01). Zur Kenntnis be Stickstoffversorgung und — bei Aspergillus niger. Ber. d. deut. bot. Ges. 19: p. (139). 190 —— ————, (09). Untersuchungen über die Stickstoffgewinnung und Eiweiss- bildung € Schimmelpilze. Beitr. z. chem. Physiol. u. Path. 2; 557-590, 1902. [See p. 559.] Fe B. M., and Knudson, L. ( Mic Relation of certain fungi to nitrogen fixa- tion. Science №. 8. 33: 191. — C. (96). Stickstofffreie aus und Enzyme. Centralbl. f. Bakt. II. 2:505-512. 1896. Frank, B. (792). Die Assimilation freien Stickstoffs bei den Pflanzen in ihrer Abhängigkeit von Species, ved EES und von Bodenarten. Landw. Jahrb. 21: 1—44. 189 — T (93). на T E Wes des freien Stickstoffs durch die Pflanzenwelt. Bot. Zeit. 51:р. 18 — Н. (708). НИ durch einige auf EEN pee üufige е Hyphomyeeten. Jahrb. f. wiss. Bot. 45; 256-302. f. 1-3. Gerlach, M., und Vogel, I. (03). Weitere Versuche mit stickstoffbindenden Bakterien, Сешта 1, f. Bakt. IT. 10: 636—643, 1903. — dE Kä (13). Can fungi living in mcm soil assimilate free nitro- gen t. Gaz. 56: 249-305. f. 1—18. Hallier, E. (67). Gührungserscheinungen. p. 22. Leipzig, 1867. [See also Zeitschr. f. Parasitenkunde 1: p. 129. 1869.] Heinze, B. (06). Sind е imstande, den elementaren Stickstoff der Luft zu verarbeiten und den Boden an Gesamtstickstoff anzureichern? An nn, Мус. 4: 41-63. 1906. ee wë 54—55. Jodin, M. ('62). Du róle physiologique de l'azote, faisant suite a un précédent travail Zonge à l'Académie dans la séance du 28 avril, 1862. Compt. rend. Acad. Paris 55: 612-615. 1862. Koch, А. (703). Bodenbakterien und Stickstofffrage. Ges. deut, naturforsch. Ärzte, Verhandl. (74 Versamml. zu Karlsbad). 1902:182-199. 1903. [See also Koch, in Lafar, Handb. d. tech. Мук. 3: 1-23. pl. 1. 1904-1906.] 1916] DUGGAR AND DAVIS—NITROGEN FIXATION 437 Kossowicz, A. (12). Die Bindung des elementaren Stickstoffs durch Saceharomy- ceten (Hefen), Monilia candida und Oidium lactis. Zeitschr. f. Gürungs- ун. 1: 253-255. 1919, -------, (14). Zur Frage der Assimilation des elementaren Stickstoffs durch Hefen und Schimmelpilze. Biochem. Zeitschr. 64: 82-85. 1914. Latham, M. E. (09). Nitrogen assimilation of Sterigmatocystis nigra and the effect of chemical stimulation. Torr. Bot. Club, Bul. 36; 235-244. Lindner, P. (12). Neue Ergebnisse bei Assimilationsversuchen mit verschiedenen Hefen und Pilzen. Chem. Zeit. 36:638. 1912. [Abs. in Myc. Centralbl. 2:24. 1913.] Lipman, C. B. (711-7112). uU нам by yeasts and other fungi. Jour. Biol. Chem. 10: 169-182, 1911-1 Lóhnis, F. (710). Handbuch der landwirtschaftlichen Bakteriologie. рр. 690-692. Berlin, 1910. Vogt ge ЗЕ (10). Beiträge sar ie ge Век Ri = rufa (Pers.). Jahrb. f. . Bot. 48; 591—631. [See pp. 625-629.1 Möller, A. (06). Mykorhizen und Stickstoffernührung. Ber. d. deut. bot. Ges. 24; 230-234, 1900. Nägeli, С. W. von (780). Ernährung der niederen Pilze durch Kohlenstoff- = А Bur enda K. Akad. d. Wiss., München, Sitzungsber. 10: p. 2 1880. Peklo, J. ('13). Neue Beiträge zur Lösung des Mykorrhizaproblems. Zeitschr. £ Gürungsphysiol, 2:pp. 275-289. 1913. pete L. H. (08). Can Fusaria assimilate free nitrogen? Mich. Acad. , Ann. Rept. 10:50. 1908. —————, (11). Upon assimilation of atmospheric nitrogen by fungi. Torr. Bot. Club, Bul. 38:135-139. 1911. Puriewitsch, K. (95). Ueber die Stickstoffassimilation bei den Schimmelpilzen. Ber. d. "deut. bot. Ges. 13:342—345. 1895. ns K. (01). Ueber die Assimilation freien Stickstoffs durch Schimmelpilze. г. d. deut. bot. Ges. 19:(107)-(115). 1901. Stahel, G. (711). Stickstoffbindung durch Pilze bei gleichzeitiger Pepe brung mit ebu ndenem Stickstoff. Jahrb. f. wiss. Bot. 49:579-615. 1911. Ternetz, C. (’04). Assimilation des atmosphürischen Stickstoffs durch einen torfbewohnenden Pilz. Ber. d. deut. bot. Ges. 22: 267-274. 1904. ——— (07). Über die Assimilation des atmosphärischen Stickstoffes durch Pilze, Jahrb. f. wiss. Bot. 44: 353-408. f. 1-6. 1907. Traaen, A. E. (14). Untersuchungen über Bodenpilze aus Norwegen. Nyt Mag. 52: 19-121. pl. 4. 1914. [See pp. 111-113.] m$ -* (12). Beitrüge zur Kenntnis der Sprosspilze ohne ENS welche Brauereibetrieben und in deren Umgebung vorkommen. Centralbl. f. Bakt. IL. 34; 1-35. 1912. [See pp. 17-22.] me. H. (09). Über eine den Luftstickstoff assimilierende Hefe: Torula Wies- К. Akad. d. Wiss., Wien, math.naturw. K1., Sitzungsber. 118: 1091- 1133. 1909 STUDIES IN THE PHYSIOLOGY OF THE FUNGI II. LENZITES SAEPIARIA FRIES, WITH SPECIAL REFERENCE ТО Enzyme ACTIVITY SANFORD M. ZELLER Research Fellow in the Henry Shaw School of Botany of Washington University* InTRODUCTION This paper reports the results of an experimental study relating to certain physiological activities of the wood- destroying fungus, Lenzites saepiaria. The investigation here reported is concerned primarily with cultural characteristics, some of the factors influencing growth and metabolism, and the enzymic activity in the fungus. Special attention is given to the eyto-hydrolyzing enzymes and the relation of these to the decay produced by Lenzites.? Tur Fuxcvus This fungus is commonly known by lumbermen as the “brown punk" because of the sepia color of the small bracket- like sporophores. In nature it is generally found on railroad ties, telephone and telegraph poles, and less often on standing timber. It attacks coniferous timber, as a rule, but is known to attack frondose wood (Weir, 714). The sporophores ap- pear near cracks in the wood due to drying. The fruiting sur- face is made up of branching gills which may become so much 1 A fellowship established by the Southern Pine Association, New Orleans, La. 2 In the fall of 1914, in coöperation with the Southern Pine Association of New Orleans, it was decided to attempt a determination of the natural factors of wood influencing in any marked degree the growth and destructive properties of wood-destroying fungi, that is, durability with respect to fungous decay. While the results in this limited field of investigation will be reported in a later paper, the present study deals with those additional physiological phases which were a necessary and fundamental part of the general plan e in search for a fungus suitable to employ in such a problem, it was deemed important to make the results as far-reaching in the economie world a possible. Since practically three-fourths of the structural timber used in the United States is furnished by the coniferous species of trees, and since Lenzites saepiaria Fries is considered the fungus most destructive to coniferous wood, this ungus was used in the investigation. ANN. Mo, Bor. GARD., VOL. 3, 1916 (439) [VoL. 3 440 ANNALS OF THE MISSOURI BOTANICAL GARDEN anastomosed that the pores formed are very much daedaloid. The mycelium is best studied in pure cultures. CULTURE RELATIONS Two methods of securing pure cultures were employed. They may be designated as (1) the tissue method, and (2) the spore method. (1) The tissue method was first described by Duggar (’05), who applied this method to the making of spawn in mush- room culture. In connection with this study on Agaricus campestris tests were made with 69 species of basidiomy- cetous fungi on various media. Forty of these grew promptly on the media employed. The method was suggested by the fact that ‘‘during moist weather, or in a moist cellar where mushrooms are being grown, one will frequently find that an injury in a young mushroom is rapidly healed by a growth of hyphae from the edges of the injured area. The same thing had been noticed in the open in the case of puffballs. In many instances, moreover, pure cultures of fungi in other groups have been obtained by small bits of a sclerotial mass of tissue.” Accordingly, young sporophores were obtained, “апа after breaking them open longitudinally a number of pieces of tissue from within were carefully removed with a sterile scalpel to a sterile Petri dish." А number of cultures were then made by transferring these nocules of tissue to various forms of nutrient media, such as bean pods, manure, leaf mould, ete. From this and from numerous other similar tests it was ascertained that when the sporophores from which the nocules of tissue were taken were young and healthy, there was seldom an instance in which growth did not result. It was shown that failure to grow was generally due to the advanced age of the sporophore used, to an un- favorable medium, or to bacterial contamination. In my work dried sporophores were used for the tissue method. These were collected in a freshly growing condition and dried at room temperature. The sporophores of Lenzites saepiaria are xerophytie, and will remain viable in a dried con- dition for some time. Buller (209, p. 111) found them to 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 441 recover after four months of desiccation, while Falck (’09) found that in one case a sporophore was still feebly viable after a year and nine months of desiccation. In my work where the tissue method was employed, the sporophores were broken open, and small pieces of the tissue from the interior were gouged out by means of a sterilized scalpel. These pieces of tissue were quickly flamed and transferred to agar slants in tubes, after dipping in sterile distilled water to moisten. The agar used was Thaxter’s glucose-potato-hard agar, made up as commonly employed in this laboratory in the following way: Two hundred grams of potato were cooked for about one hour in a liter of tap water. The potato water strained off was restored to a liter. To this were added 20 grams of glucose and 30 grams of agar, and the mixture auto- claved for 20 minutes to dissolve the agar, which was then tubed, sterilized, and slanted. This is a very good medium for the growth of Lenzites. The tissue transfers grew well, but practically one-half of them were contaminated, since diffieulty was found in secur- ing a piece of tissue from the interior of such thin sporophores without contamination. (2) The spore method was more frequently used in this work. Buller (’09) discovered the remarkable fact that many xerophytie fungi which have been preserved dry for several months or even years may be revived by moistening, when spore expulsion will be resumed and will continue for several days or weeks (according to the specificity of the organism), even after the plants have been dried and revived several times in succession. In the same year, however, Falck reported his work on the desiccation of sporophores of Len- ees saepiaria, L. abietina, and L. thermophila, in which he used this rejuvenescence due to moisture as an index of viability. The spore method is used considerably in forms with thin tissues, having been employed by Ferguson (’02), Falek (702), Lyman (’07), Münch (709), and others. My procedure differs from those of previous workers in some details, which are given here. To obtain the basidio- [Vor. 3 442 ANNALS OF THE MISSOURI BOTANICAL GARDEN spores the sporophores were rinsed twice in sterile distilled water in large-sized Petri dishes. This removes some of the bacteria and spores of foreign fungi. After this washing process the sporophores were allowed to stand in sterile dis- tilled water for about two hours, so that they were thoroughly saturated. They were removed with sterile forceps and sponged off with Scott’s tissue toweling which had been pre- viously sterilized, and were then placed, hymenium downward, in large, dry, sterile Petri dishes. After 24-48 hours the sporophores had discharged enough spores to make a white spore print. The moisture in the sporophores serves two pur- poses other than reviving the tissues. It keeps the air in the dish sufficiently humid to prevent too rapid desiccation, and it also tends to hold foreign spores to the surface of the sporophores. The latter is beneficial in securing a fairly pure dispersion of spores. The spore dispersion was made in sterile distilled water. Test-tube water-blanks were prepared and sterilized. Sev- eral loopfuls of sterile water were transferred to the spore print by means of a platinum loop. By stirring a little with the loop the spores were so dispersed that when a loopful of the spore suspension was transferred to the water-blank a cloudy streak was produced. Three or four such loopfuls of the spore suspension were transferred to a water-blank, which was well shaken by rolling between the palms of the hands. Two or three loopfuls were transferred to each of several tubes of melted agar, and poured plates were made as in the usual bacteriological method. Plates made in this way with the 3 per cent agar mentioned above were surprisingly free from bacterial contamination but contained a few scattering colonies of foreign moulds. The colonies produced by the germinating spores of Lenzites were so characteristic, how- ever, and so generally scattered over the plates that they soon became easily recognizable. Individual colonies from the plates were transferred to agar slants, a quantity of cultures being obtained. 1916] ZELLER— PHYSIOLOGY OF LENZITES SAEPIARIA 443 CHARACTERISTICS OF THE MYCELIUM IN CULTURE Lenzites saepiaria was kept in cultures on three types of media, ie. Thaxter’s glucose-potato-hard agar described above, yellow pine sawdust in Erlenmeyer flasks, and pine blocks in jars and bottles. The mycelial characters on wood are somewhat different from those on agar. Polymorphism.—Polymorphism in the Basidiomycetes has been reported by many early investigators, and in late years especially by Falck in 1902 and Lyman in 1907. Lyman reviews the literature dealing with the occurrence of oidia, chlamydospores, and conidia in certain Polyporaceae, and in his own work finds that many of the Hymenomycetes possess these several ways of reproducing vegetatively. In 1909 Falck published a monograph on Lenzites in which he de- scribes minutely the morphology and physiology of the myce- lium of L. saepiaria. The polymorphism of this fungus is extremely interesting and characteristic, the pure cultures of the mycelium being almost sufficient to identify the fungus. The mycelium is white at first, but with age the aérial part becomes a reddish brown or sepia color. When the mycelium grows out from an inoculum on agar, there is a submersed mycelium which is a forerunner of the superficial. From the latter there arises a woven mat of aérial hyphae which take on the sepia color with age. The hyphae are very much sep- tate, and clamp connections are quite common. These vary from the ordinary clamp connections through all stages to the medallion mycelium, as Falck calls it, which is found only in the wood and sawdust cultures. Falck divides the oidia into primary, secondary, and ter- tiary. The primary oidia are those formed when the whole superficial mycelium breaks up into chains of spore-like cells. The secondary are those produced at the tips of branches of the superficial mycelium. These may be abnormally swollen tips or very short chains from lateral branches. The ter- tiary oidia are the most common and appear on the aérial hyphae. The hyphae generally break at clamp connections to produce this type. The chlamydospores or gemmae are noth- ing more than swollen vegetative cells of the hyphae, which [VoL. 3 444 ANNALS OF THE MISSOURI BOTANICAL GARDEN act as conidia when isolated. In the superficial mycelium they are larger than where submersed in agar, and are often found in the medallion mycelium in the decayed wood. In the cultures on blocks of pine wood the mycelium spreads well if the blocks are fairly well saturated with water. As the moisture disappears the mycelium usually penetrates, and the superficial mycelium dries and vanishes. In this later stage of growth, however, it is difficult to see whether the blocks are well infected or whether the fungus has failed to enter and thus has dried down. In the woody tissues the hyphae extend lengthwise of the tracheids as a general rule. The medullary rays are usually full of matted hyphae. When passing from one tracheid to another the hyphae penetrate the pits. Very infrequently, however, they penetrate the walls of the spring wood, and in such cases are constricted in the tiny perfora- tion, but are swollen, forming a callus on either side of the wall. When two active hyphae come together laterally they fuse, and the whole often forms an anastomosing network. The medallions are extremely common. CHARACTERISTICS OF THE SPOROPHORES IN CULTURE Well-differentiated sporophores were formed on sawdust and pine blocks, and in a few cases sporophores occurred on agar. The earliest fruiting bodies appeared about seven months after the cultures were inoculated, and were the- lephoroid or staghorn-like in shape. The flattened projections bear the hymenium on both sides. It is composed of clavate paraphyses with the typical four-spored basidia. Still better- differentiated sporophores were produced in from eleven to twelve months, their form depending upon the surface of the substrate. Ona horizontal surface they are almost sessile and hemispherical, with the upper surface composed of tiny dae- daloid pores, while the under surface has the typical lamellate hymenium of Lenzites. When fruit bodies appear on the side of a block they tend toward the bracket form, but have hymenium above and below, the upper poroid, the lower lamel- late or hydnoid. Plate 8 shows sporophores both on sawdust and on pine block cultures. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 445 Increase in sporophore-producing capacity.—The basidio- spores from one of the sporophores produced in pure cultures were caught in a sterile Petri dish and plated out in agar as described above. There was a large per cent of germination. The pure culture tubes made from these germinated spores produced sporophores on agar in eight weeks after the trans- fers were made. This is the only instance where I secured sporophores of Leneites saepiaria on agar. It seems plausible to conclude that by growing the fungus in pure cultures the ability to produce sporophores in cultures is increased in the following generation. FACTORS INFLUENCING THE GROWTH AND METABOLISM OF THE FU In this connection four different factors are discussed. These are (1) the relation to the reaction of media, (2) tem- perature relations, (3) the water and oxygen content of the substrate, and (4) resin on wounds and in the wood. (1) The relation to the reaction of media, with special reference to chemical composition, i. e., source of carbon and other nutrients for best germination and growth, has been amply considered by Rumbold (’08), and by Falck (209) in his monograph on the Lenzites rots. Thus it was not con- sidered necessary to dwell upon these factors further than is reported elsewhere in this paper. However, in making a new supply of cultures of Lenzites saepiaria, Fomes pinicola, Polystictus hirsutus, Polyporus lucidus and others, I found that after transferring to the new medium the fungi would not grow. On testing with litmus this medium proved to be slightly alkaline. A readjustment of the reaction to slight acidity yielded a suitable medium for these forms. Rumbold (208) also found that L. saepiaria is very sensitive to alkaline media, and Spaulding (711, p. 19) found that “а number of experiments uniformly gave the same results with this species. It was found that even with one-fourth of one per cent of sulphuric acid it grew luxuriantly.’’ (2) In Falck’s (209) paper temperature relations are dis- cussed at length. He shows that L. saepiaria has a growth [VoL. 3 446 ANNALS OF THE MISSOURI BOTANICAL GARDEN range from a minimum of 5°C. to a maximum of 44°C., with 35°C. as an optimum temperature. This optimum holds for the strain of this species that I used, but the total range of temperature was not determined. A question still remains whether the optimum temperature for growth is an optimum for the complex of enzyme activities which take place during decay. With this in mind the cultures were maintained at 25-30°C., since these temperatures аге at least conducive to growth, and enzyme action in vitro is rapid within these limits. (3) Given some specific wood as a suitable substrate and a favorable temperature, then the growth of L. saepiaria will be related to such other factors as porosity, water content, oxygen tension, and abundance of stored starches or other food materials in the wood. The oxygen content of wood is necessarily related inversely to water content, provided there is water in excess of that imbibed by the tracheid walls. Vari- ation in the imbibed water must influence the degree of humidity of the air in the lumen of the tracheids, and doubt- less the humidity of this enclosed air plays a róle in the growth of the fungus through the wood. Another factor which influences the oxygen content of wood is the average size of the cell lumen. This decreases in size from the spring to the late summer wood, for in the latter the lignification in- creases the thickness of the walls at the expense of lumen capacity. High specific weight is directly related to the amount of summer growth (Johnson, '93). This necessarily means that as the specific weight of the wood increases the oxygen content would be decreased. Miinch (209) has shown by numerous experiments with various forms of wood-destroying fungi that air content is an important factor influencing the entrance of fungi. The greater number of the forms which he used have a high air requirement. The quantity of wood fibre is also important. He mentions the fact that narrow annual rings are more resistant than broad ones, because there is less capacity for air in the narrower. In specimens of wood where only some rings are decayed the decayed rings prove to be the more 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 447 porous ones. This shows that a certain undetermined mini- mum of oxygen will prevent the growth of certain fungi. Ex- ceptions to this, however, may be numerous. For instance, Münch found that Armillaria mellea is not dependent on air in the substrate. The rhizomorphs of this organism are of such a structure that they conduct air into water-saturated tissues. Doubtless there are other forms which have the power to thus conduct air to parts where there is a paucity of air. Undoubtedly, high water content will inhibit the entrance of certain fungi, but as soon as there is a paucity of water the tissues are as susceptible as ever, for there is no change in the properties of the host tissues themselves. Thus, an in- creased water content as a factor in the immunization of a host plant against disease (as Miinch would lead one to think) is not compatible with present-day ideas of immunity. Appel (715) concurs in Münch's idea that the paucity of air due to high water content may be made an effective method in the control of certain plant diseases. He applies this to die-back diseases of trees due to species of Valsa and other fungi. He says: “When such diseases occur, you will find the cause in defective irrigation methods, which may be remedied by changing the irrigation system. It is of the greatest importance that the land be irrigated at the time the trees contain less water and plenty of air, and that the next irrigations be made in time to prevent an excessive decrease of the water in the tissues.” Further, he states that the same principle may be found to be applicable to bacterial diseases of trees, especially Bacillus amylovorus, and finally remarks: “It may be possible that not only trees, but also herbaceous plants, show relations between fungous growth and air con- tent. I think it must be so for the organisms which cause the wilt diseases and the rhizoctonia disease of the potato, both of which have a high air requirement. . . . .. hough caused by a fungus, the production of conditions favorable to the progress of the disease [Rhizoctonia] is attributable to irriga- * H The writer believes that the close application of the work of Minch given by Appel to such diseases as those caused by [VoL. 3 448 ANNALS OF THE MISSOURI BOTANICAL GARDEN Bacillus amylovorus and Rhizoctonia is hardly acceptable without adequate specific data for each of the organisms concerned. Speaking of the fungous diseases in the tropics, Westerdijk (215) says: “The heavy rainfalls, combined with the abundant transpira- tion—owing to the intense heat, must cause a high water-content and a small air-content, of the wood-vessels of the trees, thereby making a substratum poor in air. This fact, combined with the high temperature, would explain the rare occurrence of LOTION and other wood-destroying fungi in the tropics A certain balance between water and oxygen is necessary, and this varies according to the specificity of the organism. Just what percentage of water in the cell walls and oxygen in the lumen of the wood fibres are necessary for the entrance of fungi are undetermined factors, but we know that both are necessary. Well-seasoned wood is very durable as long as it is kept dry. On the other hand, upon damp wood spores of fungi germinate and penetrate readily. In speaking of Len- gites spp. Falck (209, р. 223) says that the spores germinate with every rain, then there forms a small colony from which hyphae enter the wood substance. In such colonies are found the typical medallion mycelium which endures dry periods, and which after a thorough saturation with water is again able to continue its life activities undisturbed. Wehmer (714) found that bits of mycelium of Merulius lacrymans transferred to air-dried blocks would not grow at room or cellar humidity. When the blocks were well saturated with water better results were obtained for mycelial growth, but decay was not evident in all cases. As decay by Merulius spreads the moisture content increases, decayed wood showing 25 per cent hygroscopic water in damp air, where sound pine holds but 15 per cent. During the course of my work with Lenzites grown on pine blocks some interesting facts were noted concerning the growth and decay as they were influenced by the water and oxygen content of the wood. Blocks which were sterilized 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 449 without excess of water were inoculated and enough water added from time to time to keep the humidity high, but the mycelium did not spread far from the place of inoculation. Where the blocks were kept saturated until after inoculation the mycelium grew rapidly over the surface, no matter what the nature of the block. As the water evaporates the mycelium penetrates, and the superficial growth dries down and disappears. The interior of the blocks, however, holds moisture for a longer time than the surface, and in such a proportion to the wood fibre that the nearest to optimal water content is reserved in the interior. This is shown by examples of Lenzites rot wherever it is found in nature, as well as in pure cultures. Blocks that show a reduction in weight (due to decay) of from 40-60 per cent show internal decay, with a crust of fairly sound wood over the surface. If two blocks fit rather closely together during incubation the surfaces in contact may be decayed. A large series of blocks inoculated with L. saepiaria were kept saturated for a year, and they were reduced very little by the fungus. The reduction was all superficial and appeared as a ‘‘scorching’’ of the surface. Microscopic examination showed that the hyphae penetrated to a depth of but three or four tracheids. Plate 8, figs. 13-17, shows a series of blocks having this superficial ‘‘scorching,’’ and figs. 8-12 show the internal decay mentioned above. From these observations it is apparent that the oxygen requirement of L. saepiaria is low. A certain percentage of water is a necessary factor, but total saturation is injurious to the fungus because of the paucity of oxygen. The optimum, maximum, and minimum percentages of air and water have not been determined. From the above observations it will be seen that any factors influeneing the proportion of water and air are of great im- portance. Seasonal cracks, due to drying, of ties and other structural timbers afford an entrance place for the fungus and the necessary air, and usually the decay is found in radial blotches when the end of an infected timber is observed. (4) When coniferous trees are wounded in one way or another the majority of them exude pure resin from the [Vor. 3 450 ANNALS OF THE MISSOURI BOTANICAL GARDEN bark, and this seals the wounds to the exclusion of the fungus. We have found that the mycelium of L. saepiaria will not grow on 100 per cent resin plates because of the lack of nutrition, nor will the spores germinate on such a medium. In living trees, then, the pure resin covering the wounds serves a twofold purpose. It mechanically prevents the en- trance of the fungus, and excludes the air which might other- wise gain access to the interior of the tree more rapidly than under natural conditions. Resin does not exist in this high percentage, however, in the interior of wood, but is infiltrated into the lignified walls. Hence, there is still air in the lumen and food accessible to the fungus. Laboratory experiments reported below have shown that under these conditions the presence of resin has very little influence upon the growth of the fungus, at least up to 50 per cent resin by weight, which is considerably more than is found in any coniferous wood. A quantity of resin was extracted by means of benzol from longleaf pine wood (Pinus palustris). The resin was hard- ened at 65°С. until it was of a constant weight. This was used in making a resin agar, the basis of which was a 4 per cent Thaxter’s glucose-potato-hard agar. The powdered resin was added to the agar while warm, emulsified by vigor- ous agitation, and then sterilized. Sterilization may be done in the Arnold sterilizer on three successive days, or if it is necessary to sterilize but once, autoclaving is satisfactory; but when autoclaved more than once the resin seems to be acid enough to hydrolyze the agar sufficiently to keep it from hardening. If the agar is removed from the autoclave while it is still quite super-heated and vigorously agitated while it is cooled by placing under the water tap from time to time, a very satisfactory emulsion of any percentage may be obtained. The agar was made up so as to contain 5, 10, 15, 20, ete., up to 100 per cent, of resin, which was plated out and after cooling was inoculated with squares of mycelium of L. saepiaria cut as suggested by Humphrey and Fleming (715, pl. 1). The inocula were 0.8 em. on a side. After 14 days of growth at 32° C. measurements of the diameters of the grow- 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 451 ing mycelial colonies were taken. Plate 9 shows photographs of the comparative growths. The accompanying table (table I) shows that on the 5 per cent resin agar the growth was reduced somewhat, as compared with the control containing no resin; but the growth was just about as rapid on 40, 45, and RESULTS OF GROWING LENZITES SAEPIARIA ON PLATES OF RESIN AGAR Diameter of growth on resin agar plates Per e: after 14 days at 32?C. o resin Plate Plate Plate Plate no. 1 no. 2 no. no. 4 Averages cm. cm. cm. cm. cm. ) (control) 9.0 9.0 9.0 9.0 9.0* 7.6 7.4 6.8 [9 7.42 ) 6.1 6.2 6.7 7.4 6.6 6.8 6.5 4,2 5.5 6.5 20 6.2 6.1 5.9 6.2 6.1 : 6.4 6.8 6.9 6.4 0.63 30 6.2 5.7 6.2 5.9 6.0 1 6.8 7.0 6.6 6.8 6.8 40 6.5 6.2 6.6 6.4 6.4 ¿ .4 7.0 6.6 6.4 6.85 50 $.9 6.2 5.7 6.0 5.95 3.4 3.5 3.8 3.7 3.6 60 9 1.5 1.8 4 2 ( .8 1.4 1.7 T. .6 ) .8 1.6 1.7 9 .6 7 .6 1.7 1.8 7 80 4 1.5 1.1 52 iJ 8 5 1.6 1.4 1.4 5 90 0.8 0.8 0.9 1.1 0.9 9 0.8 0.8 0.8 122 0.9 100 0.8 0.8 0.8 0.8 0.8 * The diameter of the inoculum іп each case was 0.8 cm., which should be subtracted from the above values in order to obtain the actual growth. 50 per cent resin as on 5, 10, and 15 per eent, the average growth of the former three being 6.4 em., and of the latter 6.84 em. From 50 to 60 per cent there was an abrupt change in the growth, which showed marked inhibition from 60 to 85 per cent, while above this there was practically no growth. The fact that there was some growth up to 85 per cent resin is con- clusive evidence that resin is not toxic. Of course, it could not be toxic unless soluble in water or the enzymic fluids excreted by the fungus. Its greatest inhibitive power lies in the fact that it excludes water from the substrate. The results given [Vor. 3 452 ANNALS OF THE MISSOURI BOTANICAL GARDEN here are merely indicative of the true conditions as they exist in nature, since the nutrition in the two cases is different. We know from the work done by Le Renard (712) that the composition of the nutrient medium used in such Petri dish plates has a marked influence on the effect of any toxic sub- stance present; that is, if the nutritive elements present are varied in quantity there is a change in the effect of the toxic substance on the growth of the organism. Resin determinations have been made on approximately 450 series of pine blocks, the whole comprising about 3000 samples which were placed in cultures of L. saepiaria. The reduction in weight of these blocks, after incubation for one year, varies considerably, but the results so far taken tend to show that the influence of resin on the decay by this fungus is exceedingly erratic. This will be reported in a second paper. ENZYME Activiry IN LENZITES SAEPIARIA Work on the enzyme activity of the wood-destroying fungi is comparatively meagre. This is especially the status of the cytolytic investigations. There are very few papers wholly devoted to enzymes of higher fungi. In 1895 Bourquelot and Hérissey investigated the enzymes from the juice of the sporo- phores of Polyporus sulphureus. The enzymes were precipi- tated with alcohol. Czapek, in 1899, found in natural infec- tions of Merulius lacrymans an active principle capable of liberating from lignin the substance which gives the lignin reactions in alcoholic extracts. This substance, which is dis- cussed more fully later in this paper, he called ‘‘hadromal,’’ and the enzyme capable of liberating it was called ‘‘had- гошаве.””. Two years later Kohnstamm (201) applied Buchner’s ‘‘Dauerhefe’’ method to Ше sporophores and mycelium of Merulius lacrymans and Armillaria mellea. He obtained evi- dence of the presence in these fungi of diastatic, proteolytic, glucoside-splitting, and cellulose-hydrolyzing enzymes. Buller (206) tested out the juice expressed from the sporophores of Polyporus squamosus and obtained positive evidence of the 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 453 presence of eight enzymes. Reed (?13) grew Glomerella rufomaculans in cultures of nutrient solutions, and from the dried fungous mat was prepared a fine, enzyme-containing meal which was tested on various substrates. The few papers mentioned are the main ones dealing en- tirely with enzyme activity in fungi which attack wood. Many other investigators have inferred a priori that many enzymes, especially cytolytic, are active agents in the metabolism of this group of fungi, and scattering references of this sort are numerous. It is of interest to observe that although we have every indication to direct us to believe that cytases are present in wood-destroying fungi, yet their presence has been demon- strated only indirectly, i.e., by histological methods. This may be due to the fact, as will be pointed out later, that a majority of the investigators who found no wood-destroying ferment used the fruiting bodies in their experiments instead of the active, vegetative mycelium. More detailed discussions of the results of these various investigators will be given below in connection with the various groups of enzymes considered in this paper. In view of the status of our knowledge of the enzymes con- cerned in the destruction of wood, I undertook an investiga- tion to compare in a qualitative way the enzymes of the mycelium and sporophores of Lenzites saepiaria. In certain cases only have quantitative results been obtained. METHOD OF GROWING MYCELIUM FOR EXTRACTION Sawdust of Pinus palustris, P. echinata, and P. Taeda was placed in flasks of 1000 and 500 ce. capacity. The sawdust was moistened with distilled water, after which the flasks were plugged and sterilized in the autoclave at 20 pounds pressure for 45 minutes. After cooling, the sawdust was inoculated with the mycelium from agar slants. As the mycelium grew into the sawdust there was a darkening of the wood similar to that noticed in the ‘‘rot’’ produced in nature by this fungus. In the course of time the sawdust became a dark brown color. After about 7 months some of the flasks were emptied, and the sawdust with the mycelium was dried by means of an [VoL. 3 454 ANNALS OF THE MISSOURI BOTANICAL GARDEN electric fan blowing a draft of warm air (about 30°C.) over it. After being dried the sawdust could be crushed to powder between the fingers. This dry sawdust was ground to a very fine powder in an ordinary mill. A tared amount of the powder was transferred to a clean liter flask, and about 4 parts of water by volume were added with about 1 per cent chloro- form. This was allowed to stand for 16 hours for the extrac- tion of enzymes. Then the solution was filtered off through a Buchner funnel, and the enzymes precipitated from the filtrate by the addition of 3 volumes of 95 per cent alcohol. The pre- cipitate was collected on a filter paper in a Buchner funnel, and the paper allowed to dry at room temperature. These filter papers were kept in glass-stoppered bottles for future use. When the enzyme preparation was to be used the filter paper was soaked in such a quantity of water that each cubic centimeter of the resulting enzyme ‘‘dispersion’’ was equiva- lent to 1.5 grams of the original sawdust powder. In the fol- lowing work 2 ce. of this dispersion were used in 10 се. of the respective substrates. PREPARATION OF SPOROPHORES FOR EXTRACTION Sporophores of Lenzites saepiaria were collected by the writer at Leeper, Missouri. Only the viable, light sepia- colored specimens were used in the enzyme work. The tissue of the sporophores, after grinding, was treated in the same way as described above for the mycelium in sawdust. The enzyme preparation was secured in the same way, and 1 се. of the enzyme dispersion in water had the value of 1.5 grams of the original sporophoral meal. Unless otherwise specified in the following pages ‘‘mycelial meal" will refer to the original powdered sawdust including mycelium before extrae- tion, ‘‘sporophoral meal," the original ground sporophores before extraction, ‘‘mycelial dispersion," the dispersion of enzymes extracted from the mycelial meal, and ‘‘sporophoral dispersion,’’ the dispersion of enzymes extracted from the sporophoral meal. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 455 ENZYMES OF LENZITES SAEPIARIA E RASES By the aetion of esterases the esters of the fatty acids are saponified and are thus resolved into their constituents, aleohols and fatty acids. It is thus possible to recognize and measure the activity of the enzymes in decomposing esters by determining the acidity of the substrate quantitatively by titration against alkali. The presence of fatty globules in the hyphae of fungi has led to the investigation of the enzymes capable of accom- plishing their hydrolysis. Biffen (799) found а fat-destroying fungus belonging to the Hypocreales which grew luxuriantly on the endosperm and milk of the cocoanut. All cultures of the fungus showed that the fats became emulsified, and the substrate became increasingly acid with continued growth and had a pleasant ethereal odor something like that of amyl butyrate. On triturating the mycelium with kieselguhr, and filtering under pressure, he obtained an extraet which decom- posed both cocoanut oil and monobutyrin. Buller (’06) found that 10 ce. of the juice from the sporophores of Polyporus squamosus hydrolyzed 43 per cent of a 1.84 per cent ethyl ace- tate solution in 330 hours. He mentions “that when spores of Polyporus squamosus are allowed to dry for several days, many of them develop large fat drops. On germination of the spores in malt-wort extract these drops disappear. Per- haps this is due to the action of lipase." Bayliss (208) was not able to demonstrate the presence of lipase in Polystictus versicolor. Dox (710) has reviewed the literature concerning the filamentous fungi which split the fats. Reed (713) found that the enzyme powder prepared from Glomerella rufomacu- lans split ethyl acetate and ethyl butyrate, but that the in- erease in acidity was much greater where ethyl acetate was used as the substrate. The presence of oil globules in the spores and mycelium of L. saepiaria led to the investigation of the power of the fungus to utilize such substances. Thus, experiments were conducted using as substrates the following: olive oil emul- [VoL. 3 456 ANNALS OF THE MISSOURI BOTANICAL GARDEN sion, triacetin, methyl acetate, ethyl acetate, and ethyl butyrate. Olive oil emulsion was made according to Bloor’s (714) method, which was previously reported from this laboratory by Davis (215). Ten cubic centimeters of olive oil were dis- solved in hot, absolute alcohol. This was run through a hot funnel to which was attached a piece of glass tubing drawn out to a very fine jet. The fine stream of oil in alcohol was run into 100 ce. of cold distilled water, which was constantly stirred. The milk-white emulsion was finally boiled to drive off the alcohol, then was diluted to 500 ec. with distilled water. The other substrates were made up in 1 per cent solutions, and if kept any length of time toluol was added as an anti- septic. All substrates were used according to the following example and were always set up in duplicate: (1) 25 се. ethyl acetate + 5 cc. enzyme dispersion + toluol. (2) 25 се. ethyl acetate +- 5 ee. enzyme dispersion (auto- claved) + toluol. (3) 25 се. ethyl acetate +- 5 се. water + toluol. (4) 25 се. ethyl acetate + equivalent weight meal + toluol. (5) 25 се. ethyl acetate + equivalent weight meal (auto- claved) + toluol. The results show that the lipolytic action on neutral fats is very slight, if any, but that on the esters of the lower fatty acids it is more marked. As Reed (713) observed for Glom- erella rufomaculans, here the acetates are better substrates than the butyrates. Methyl acetate is more strongly hydro- lyzed than ethyl acetate. Table п shows the results of the action of the esterases in the mycelial meal and the enzyme dispersion from the myce- lium. These cultures were incubated for 24 days at a tem- perature of 25-30°C. Table m shows the lipolytic action in the sporophores under the same conditions. Since the mycelial meal includes the sawdust upon which the mycelium grew, the results obtained for the mycelium and sporophores are not comparable, but from the results on methyl acetate it can be readily seen that the esterase activity in the mycelium is stronger than in the sporophores. 1916] ZELLER— PHYSIOLOGY OF LENZITES SAEPIARIA TABLE II 457 ACTION OF ESTERASES OF THE MYCELIUM UPON VARIOUS SUBSTRATES Number cc. N/20 NaOH to Form of enzyme neutralize 10 cc. substrate Substrate material Gross Net Methyl acetate. ..| Enzyme dispersion............. 2.95 2.30 Meth З Enzyme dispers E (autoclaved).. DE, E Ethyl acetate..... Enzyme dispersion. ............ 1.60 0.70 Ethyl acetate..... En e disper (autoclaved) ШЕСІ. Ethyl butyrate ...| Enzyme dispersion............. 0.15 0.05 Ethyl butyrate ...| Enzyme di (autoclaved) ОТО DENT. 522. Olive oilemulsion..| Enz dispersion............. 0.35 0.10 Olive oil emulsion..| Enzyme dispers s c E ТЕРІСІНЕ еее o Is Olive oil emulsion. . ent weight of meal....... GH 0.07 Olive oil emulsion.. Equivalent weight of bed (auto- T ps) ne "Seege? weight of meal....... 9.08 0.48 THEN. ара ъс Equivalent weight of est (auto- EE OMA EE EM DX EEN TABLE III ACTION OF ESTERASES OF THE SPOROPHORES UPON VARIOUS SUBSTRATES Number cc. N/20 NaOH to neutralize 10 cc. substrate Form of enzyme Substrate materna Gross Net Methyl acetate. . .| Enzyme dispersion............. 7.10 2.30 Methyl acetate. . .| Enzyme dispersion n моба», "MEM OT CS S. Methyl acetate. . .| Equivalent wt. meal............ 12.30 5.90 ethyl acetate... cud ыз mei (autoclaved)| - Ж Рече: Ethyl acetate..... Enzyme dispersion............. 4.55 1.25 Ethyl acetate..... К dispersion (autoclaved).. c Me Rae Ethyl acetate..... Equivalent wt. meal............ 9.40 4.70 Ethyl acetate..... Equivalent wt. meal (autoclaved) Se: ee Ethyl butyrate... .| Enzyme dispersion............. 0.42 0.07 Ethyl butyrate....| Enzyme debeo (autoclaved).. PEE Bol... Ethyl butyrate... .| Equivalent wt. meal............ 2.65 0.95 Ethyl butyrate... .| Equivalent wt. mg (autoclaved) — Ж фал. 7T MP Olive oil emulsion. .| Enzyme dispersion. ............ 1.10 0.25 Olive oil lsi nzyme aa (autoclaved) EE. EE MALTASE Of the disaccharases, I tested for maltase, lactase, and in- vertase. The occurrence of maltase in some of the lower fungi is of special interest and importance, but the literature has been adequately discussed by Dox (210). fungi it has been demonstrated in the sporophores of Polyp- In the higher [VoL. 3 458 ANNALS OF THE MISSOURI BOTANICAL GARDEN orus sulphureus by Bourquelot and Hérissey (’95), and by the writer it has been found both in the mycelium and sporo- phores of L. saepiaria. In our experimental work a 1 per cent solution of maltose was used as a substrate. Ten cubic centimeters of this were placed in each of 6 test-tubes. To each of 2 of these were added 2 ec. of the mycelial dispersion, to each of 2 others, TABLE IV SHOWING THE ACTION OF MALTASE IN LENZITES SAEPIARIA AFTER TWO WEEKS AT 25-30*C. Reducing sugars as Net due to Amount of 5:2 877 glucose іп 5 cc. hydrolysis in 10 cc. of 1 per of substrate by maltase Mycelium | Sporophore | Mycelium | Sporophore g. mg. DE bc IUe eerie 40.40 39.24 6.28 18.61 2 es (agtociaved); oorr 34.12 20.63 | у... у СООЗ WOR ual rA XS 26.16 10-72- Ñ+ 5e bMS 2 ec. of the mycelial dispersion which had been autoclaved to kill the enzymes, and to each of the remaining were added 2 сс. of distilled water. To all were added a few drops of toluol as an antiseptic. A comparable series of experiments was set up using sporophoral dispersion. All were ineubated at 25-30°C. for 2 weeks, after which time they were tested for reducing sugars by Shaffer's method ('14). Maltase reduces Fehling's solution, but when it is hydrolyzed each molecule yields 2 of glucose which would reduce almost twice as much Fehling's solution. Table 1v shows that there was consider- ably more reduction in the ‘‘regular’’ tubes than in the auto- claved and the water controls. The fact that there is more net reduction due to maltase in the sporophores than in the mycelium is not of any significance when the amount of saw- dust in the mycelial powder is considered. LACTASE Laetase has never been reported from the higher fungi. In dealing with laetase the same procedure was carried out 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 459 as for maltase, except that a 1 per cent solution of lactose was used for a substrate. There was no indication of the ` presence of lactase either in the mycelium or the sporophores. INVERTASE Although invertase has been repeatedly demonstrated in lower fungi, especially yeasts, Aspergillus and Penicillium, its presence has seldom been noted in the higher forms. Bayliss (708) found it in the sporophores of Polystictus versicolor, and it is undoubtedly present both in the mycelium and sporo- phores of L. saepiaria. To demonstrate the presence of invertase a 1 per cent solu- tion of sucrose was used as a substrate. To 10-сс. portions of this were added 2 сс. of the enzyme dispersions, enzyme dispersions autoclaved, and 2 cc. of distilled water, as in the maltase experiments. After 4 hours there was distinct reduc- tion of copper oxide from Fehling’s solution in the regular tubes, but the autoclaved controls and the water controls showed none. This was perhaps more evident in the sporo- phoral dispersion. Having demonstrated the presence of invertase both in the mycelium and sporophores, some quantitative studies were made. The fungus was successfully grown upon a substrate of carrot juice. The carrot juice cultures were made in Erlen- meyer flasks and inoculated with oidia from cultures grown on pine sawdust. The oidia were dispersed in sterile, distilled water-blanks from which the inoculations were made. After 2 weeks of growth the mats of mycelium were removed from the flasks and dried on filter paper at a temperature of about 35°C. This dried mycelium was ground to a fine powder in a mill and kept dry in a glass-stoppered bottle. 'This same powder was used for quantitative determinations of diastatie aetion reported later in this paper. The experiments were conducted as follows: To 50 cc. of a 1 per cent sucrose solution 2 grams of fungous powder were added, with about 1 per cent toluol as an antiseptic. As а control on this, other experiments were prepared in the same way after the fungous powder had been autoclaved to kill the [VoL. 3 460 ANNALS OF THE MISSOURI BOTANICAL GARDEN enzyme. The sucrose solution was also used alone. These were all set up in duplicate, and at the same time parallel experiments were set up with sporophoral meal in the same way. They were allowed to remain at room temperature (about 27°C.). After a period of 3 hours one set of cultures was killed in the autoclave by allowing the pressure to come to 10 pounds. The duplicate set remained in incubation for TABLE V QUANTITATIVE STUDY ei THE INVERTASE ACTIVITY IN MYCELIUM D SPOROPHORES Reducing sugars as glucose Net invert sugar due to in 10 cc. of substrate invertase activity Amount of fungous | After 3 hrs. | After 6 hrs. | After 3 hrs. | After 6 hrs. powder in 50 сс. of 1 | incubation | incubation incubation | incubation per cent sucrose solu- n Ф Ф Ф Ф Е 8 Е E Е 8 Е E 3181318/318] ЕК See ee HERR ai = A = 27 = A = N mg. | mg. | mg. | mg mg. | mg. | mg. | mg. A WOU. SS ca ee IT 7.52 | 2.88 |17.19| 5.16 | 5.44 | 2.25 115.05] 4.53 2 grams (autoclaved)... .| 2.08 | 0.63 | 2.14] 0.63 |......]......]......]...... Sucrose solution alone. . .| Negli- Negli- gible |...... Са А TET METEO КЕСИК с Mrs 6 hours and then was killed in the same way. After killing the substrates were filtered, and the invert sugars were de- termined by the Shaffer method. Table v gives these quanti- tative results. "These results show definitely that the greater invertase activity is in the vegetative part of the fungus, the inversion taking place over twice as rapidly in the mycelium as in the sporophores. In collecting such sessile sporophores as those of L. saepiaria it is diffieult at times to remove the fruiting body without taking with it some superficial myce- lium. It may be that if only the marginal portions of the sporophores were used in this work there would be a greater difference in the activity between fruiting body and mycelium. RAFFINASE A earbohydrase which transforms the trisaccharide raf- finose into fructose and melibiose was demonstrated. To 10 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 461 ec. of a 1 per cent solution of raffinose were added 2 ce. of enzyme dispersion and toluol as an antiseptic. Controls were prepared as in previous experiments. After 48 hours Feh- ling’s solution was strongly reduced in all but the controls. This was true both for the enzyme preparations from the mycelium and the sporophores. This gives evidence of the presence of raffinase in L. saepiaria. Dox (710) has reviewed the literature as to the occurrence of this ferment in Aspergillus and Penicillium. He found that mould powder of Penicillium Camemberti hydrolyzed raf- finose, and that varying the source of carbon in the substrate exerted an influence on the amount of raffinase produced. A significant fact brought out is that lactose and sucrose yielded a larger quantity of raffinase than did other carbohydrates, and these two disaccharides, it is to be noted, contain two of the hexoses found in raffinose; that is, lactose on hydroly- sis yields galactose and dextrose, and sucrose yields dextrose and levulose. | The presence of raffinase in higher fungi has not been demonstrated before, as far as the author is aware. EMULSIN The presence in plants of an enzyme capable of decom- posing glucosides has been known since 1837, and emulsin was discovered in fungi in 1893 by Bourquelot, who found it in Aspergillus niger, and by Gerard, who found it in Pen- icillium glaucum. Bourquelot (94) was able to detect emulsin in many of the higher fungi found on wood. Among those tested, 34 species (mostly Basidiomycetes) showed the pres- ence of emulsin, and 9 did not. None of the 9 were found on wood. It is probable that in the destruction of wood, whether frondose or coniferous, glucosides are set free. Among these are salicin, populin, arbutin, and amygdalin from fron- dose woods, and principally coniferin from the conifers. Be- fore these are available as nutrients for the attacking fungus they must be acted on by emulsin, which splits the glucoside, yielding glucose which is directly assimilable. [VoL. 8 462 ANNALS OF THE MISSOURI BOTANICAL GARDEN In 1895 Bourquelot and Hérissey found that the juice ex- tracted from the sporophores of Polyporus sulphureus actively digested the glucosides, arbutin, amygdalin, aesculin, con- iferin, and salicin. Working on Armillaria mellea, Merulius lacrymans, and Polyporus squamosus, Kohnstamm (201) showed that emulsin is present in the sporophores as well as in an extract of the wood decayed by these organisms. Buller (206) found the expressed juice of the sporophores of Poly- porus squamosus to act similarly toward amygdalin, while Bayliss (’08) reported that negative results were obtained using an extract of Polystictus versicolor. Reed ('13) re- ported it from the mycelium of Glomerella rufomaculans. In my experiments a 1 per cent solution of amygdalin was used as a substrate. Ten cubic-centimeter portions of this were placed in test-tubes, and 2 cc. of the enzyme dispersion were added to 3 of these and 1 was boiled. In another, 10 ce. of the amygdalin solution were diluted with 2 со. of distilled water as a control. To all was added toluol as an antiseptic. All were incubated at 25-30°C. for 3 days. After incubation the two regulars reduced Fehling’s solution, gave a strong odor of benzaldehyde and the Prussian blue test for hydro- cyanic acid. The boiled control and water control gave none of these tests. The Prussian blue test was not quite as pro- nounced in the sporophoral as in the mycelial material. The biological importance of the presence of emulsin in L. saepiaria is interesting, since we know that the pine, upon which the fungus grows most readily, contains coniferin. When coniferin is hydrolyzed by emulsin it yields glucose and coniferyl alcohol. The latter, through the action of oxidases, yields vanillin. Glucose is thus made available by the action of emulsin. Blocks of wood which had been in cultures for one year, during which period they were always saturated with water, were decayed only over the surface. When these were dried in an oven at 65°C. erystals of vanillin were collected over the surface of the blocks and the interior of, as well as about, the apertures of the oven. When the blocks were decayed under moderate moisture conditions no such sublimation of 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 463 vanillin was observed. Water then may be a factor to increase this oxidation process in the production of vanillin from woody tissues; at least very moist conditions seem to aid this action or the action of emulsin in the liberation of the coni- feryl alcohol. TANNASE Knudson (713, *13*), in his two papers on the tannic acid fermentation, gives a review of the literature on this subject up to that time. All of this literature deals with the tannase found in Aspergillus and Penicillium, but no work has been TABLE VI STUDY OF THE HYDROLYSIS ES TANNIN BY THE ENZYME DISPERSION FROM NZITES SAEPIARIA Gallic acid after 4 Net over det of enzyme dispersion in eeks’ hydrolysis Чака 2 of 1 per cent tannin solu- tion Mycelium | Sporophore | Mycelium | Sporophore mg. mg. mg. mg. 5 сс. dispersion. ............... 7.39 5.88 Om cL. LO URP 5 сс. mmer Suey E 3.28 ees 5 cc. distilled eater. 1721 EE ЖОБАНТ done to determine tannase іп the higher fungi. Knudson, however, did determine the toxicity of tannic acid for fungi, and included in his list such higher forms as Polyporus sul- phureus, P. resinosus, and Fomes applanatus. On 0.25 per cent tannic acid in bean decoction he found Polyporus sul- phureus and P. resinosus to grow well, but no growth was made by any of these forms on 2 per cent tannin. To determine the hydrolysis of tannic acid to gallic acid, Jean’s (200) iodine titration method was used. The tannin is precipitated with albumin and salted out with excess of sodium chloride, and the gallic acid remaining is titrated against a standard iodine solution. The value of the albumin solution in terms of the iodine must be subtracted from the gross value of the titration. The experiments follow in table vr. The increase of gallic acid here is very strong and must be regarded as evidence of tannase in the mycelium of the fungus. [VoL. 3 464 ANNALS OF THE MISSOURI BOTANICAL GARDEN DIASTASE Hartig (’78) was the first to mention the digestion of starch by a wood-destroying fungus, Fomes annosus, but not until the work of Bourquelot (’93-’96) did we know that diastase is widely distributed in these higher fungi. Bourquelot’s work was especially with the sporophores of Polyporus sul- phureus. Kohnstamm (’00) demonstrated the presence of diastase in Merulius lacrymans, Polyporus squamosus, and Armillaria mellea, and in 1906 Buller verified this work with the juice from the sporophores of Polyporus squamosus. Diastase was found present in both the mycelium and sporo- phores of L. saepiaria. Potato starch was made up into a 1 per cent paste in the manner which is commonly used in this laboratory in the advanced plant physiology course conducted by Professor Duggar. Two and one-half grams of potato starch in 150 ce. of distilled water were brought to boiling, while constantly stirred. This was transferred to a flask containing about 600 ec. of hot distilled water. The whole was boiled in a reflux condenser for about 2 hours. After cooling the paste was made up to a liter by adding distilled water. To the soluble starch thus prepared about 1 per cent toluol was added as an antiseptic. A series of experiments was set up as for maltase but using the starch paste as a substrate. After 4 hours the mycelial dispersion showed decided indication of the presence of reduc- ing sugars, while the sporophoral dispersion after 8 hours of incubation showed comparatively less. This work was not quantitative, however, and later experiments were conducted so that quantitative results could be taken. For this work I used the fungous powder prepared from mycelium grown on carrot juice, described under invertase. The fungous powder was used directly without extracting the enzymes. А 1 рег cent starch paste was made as above described and this used for the substrate. In each experiment 50 ce. of the paste were used. To this were added 2 grams of the fungous powder and enough toluol to act as an antiseptic. Control experiments were set up in which the fungous powder had been autoclaved. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 465 Parallel experiments were prepared using the ground sporo- phores. The results of these experiments are incorporated in table уп. As was the case with the invertase, the diastatic activity shows up much more strongly in the mycelium than in the sporophores. TABLE VII QUANTITATIVE STUDY OF THE DIASTATIC ACTIVITY IN MYCELIUM AND SPOROPHORES Reducing sugars as glucose Net due to hydrolysis in 10 cc. of substrate y diastase After 3 hrs. | After 6 hrs. | After 3 hrs. | After 6 hrs. Amount of fungous incubation | incubation | incubation | incubation powder in 50 cc. of 1 per cent starch paste Ф Ф Ф Ф ELS [28 | 8 |a е s= Е & | Е a | Е & | 5 & Š 2 5 - Š S 8 р E E & ^ & T & = a = 27 = Ф = г mg. | mg mg mg. | mg mg. | mg. | mg. ams powder........ 10.67 | 3.18 | 24.68] 7.96 | 8.50 | 2.39 | 22.54] 7.20 2 о» lebt зы (auto- e 17 79 | 2.14 (LEE EE, EE BEA Starch paste alone...... Negli- Negli- е Gs ео. E ER CYTO-HYDROLYZING ENZYMES Under the general term cyto-hydrolyzing enzymes, I shall consider all enzymes which attack such higher carbohydrates as lignin, cellulose, hemicellulose, and pectic bodies. In the succeeding pages the following classification of this group of enzymes will be used: 1. Ligninase, called ‘‘hadromase’’ by Czapek (799), to designate the enzyme capable of splitting lignin. 2. Cellulase, the true cellulose-hydrolyzing ferment. 3. Hemicellulase, the ferment hydrolyzing the hemicellu- loses. 4. Pectase, the enzyme capable of clotting the pectins. 9. Pectinase, an enzyme which hydrolyzes into reducing sugars the pectinous substances, especially the middle lamellae of plant tissues. [VoL. 3 466 ANNALS OF THE MISSOURI BOTANICAL GARDEN Ligninase.—It has often been sought to determine compre- hensively the chemical composition of the non-cellulose com- ponent of woody membranes. Tiemann and Haarmann, in 1874 (cited from Grafe, '04), believed this component to be coniferin, while Singer (’82) considered lignin as a mixture of coniferin, vanillin, and wood gum, which gave a test for the aromatic aldehydes. The prompt action of Schiff’s aldehyde reagent with rose aniline and sulphurous acid speaks for the occurrence of aldehyde-like substances in lignin. In fact, in 1898, Czapek succeeded in splitting off a substance from lignin by cooking it in stannous chloride solution. This sub- stance gave the typical wood reaction when treated with phloroglucin and hydrochloric acid, and was described by him as an aromatic aldehyde which he called hadromal. Accord- ing to Grafe (704), Czapek’s hadromal does not act like a homogeneous body but like a mixture of vanillin, methyl fur- furol, catechol, and coniferin, which substances exist in the form of an ether-like compound with the cellulose of the cell wall, or are taken up by resin, or may be found free in slight amounts in the wood fibre. According to Czapek (713), how- ever, catechol and vanillin may be regarded as decomposition products of hadromal. Other authors go only so far as to state that the substances making up lignin are intimately related to colloidal sub- stances, and can exist neither as a chemical compound with cellulose nor as its transformed product. On the other hand, Cross and Bevan (’01) hold that lignocellulose (lignin) is a complex of normal cellulose with two bodies, the one a fur- furol-yielding group, the other an aromatic or benzenoid group. Thus the chemistry of the lignocelluloses is such an open question that the decomposition products produced by enzymes from fungi are still worthy of attention. Our knowledge of the decay of wood induced by fungi prac- tically began with the fundamental researches of Hartig (’78) who has furnished extensive data concerning the parasitic and saprophytic fungi destroying the most important species of wood. He has shown that a radical change in the lignified walls is wrought by the fungus, and that in the first stages of 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 467 decay the wood gives a blue color with zinc chloriodid, after which there is a maceration or loosening of the affected walls. That the penetration of the walls by the hyphae is the result of the excretion of active fluids by the fungus was also brought out in this work. Not only do basidiomycetous fungi attack the lignified walls, but certain filamentous fungi cultivated on wood will pene- trate. Miyoshi (’95) found that Penicillium and Botrytis penetrated the tracheids of the coniferous wood by boring through the bordered pits, while Marshall Ward (’98) showed that by growing Penicillium in pure culture on blocks of spruce, the fungus could bore deep into the wood by follow- ing the medullary rays in which there was reserve starch. After these more easily assimilable foods are used up the membranes themselves are attacked. Czapek (799%) made similar observations simultaneously with Ward. Hartig ob- served that the starches disappeared very soon in the presence of the mycelium, as compared with the dissolution of the lignin, which becomes the predominant activity of the fungus. Czapek (’99*) observed that, with alcohol or benzol, a great mass of hadromal ean be extraeted directly from wood which is destroyed by the inroads of the mycelium of Merulius lacrymans, as well as from the wood penetrated by the mycelia of Polyporus adustus, Pleurotus pulmonarius, P. ornatus, and Armillaria mellea. From sound wood he obtained relatively little hadromal. The alcoholic extract from the decayed wood gives an exceedingly intense red color with phloroglucin acidi- fied with hydrochloric acid. This hadromal test is a perma- nent thing in all stages of the decay. The test for cellulose by the zine chloriodid begins to appear before the dissolu- tion of the membrane. Czapek concludes from this that through the action of the fungus the cellulose-hadromal ether is broken, and the cellulose and the hadromal are free to give their individual reactions. To demonstrate that this activity is enzymic, Czapek prepared an extract of the mycelium of Merulius lacrymans and Pleurotus pulmonarius from natural cultures. Shavings in this extract were incubated at 28°C. There was a gradual action, and after 14 days an alcoholic [VoL. 3 468 ANNALS OF THE MISSOURI BOTANICAL GARDEN extract of these shavings gave a strong hadromal test with phloroglucin and hydrochloric acid. The extracted wood gave the purple reaction with chloriodid of zinc. The fungous extract lost its activity when boiled. It could be precipitated with alcohol and thus yielded a white powder. He called the active principle ‘‘hadromase.’’ Von Schrenk (200, p. 12) isolated the same enzyme from the mycelium of Polyporus subacidus growing in spruce wood. After taking into consideration the different types of wood decay, it would seem that ‘‘hadromase’’ is a misnomer, since it does not act upon hadromal but upon lignin. In some forms of decay, such as the action of Trametes Pini upon pine (see Hartig, '78, p. 36, and von Schrenk, '00*), the white rot of the red cedar produced by Polyporus juniperinus (see von Schrenk, ’00, p. 9), and the action of Thelephora perdix on the oak, as reported by Helbig (’11) the hadromal and other bodies are split up and used by the causal organism, leaving pure white cellulose. Should an enzyme which acts on hadromal, or the soluble substances giving this red phloro- glucin test ever be isolated in these cases, it would lead to a confusion in nomenclature. It is proposed that ‘‘ligninase’’ be used to designate the enzyme capable of splitting lignin. A lengthy list of papers may be cited which deal with timber-destroying fungi and which refer in a direct or indirect way to the lignin-splitting enzyme. Among these publications which have not been cited above may be mentioned the work of Biffen (’01) on the biology of Bulgaria polymorpha, of Marshall Ward (’97) on cultures of Stereum hirsutum, of Buller (705) on the destruction of paving blocks by Lentinus lepideus Fr., and (’06) Polyporus squamosus as a wood de- stroyer, of Falck (709, 719) on the dry rots of Lenzites and Merulius, of Wehmer (712, 714), and various papers by von Schrenk (200, 700%, 200, 201, '03, 14, 14). Whether the authors mentioned here have isolated the enzyme or not, it is probable a priori that the lignin-splitting enzyme is present in the fungi with which these papers deal. Since Lenzites saepiaria produces a typical brown rot sim- ilar to that produced by other dry rot fungi, such as Merulius 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 469 lacrymans, I was interested in the stages of decay and the enzymes involved in the destruction of the wood. In addition to the study of the enzymes, observations were made on the microchemical reactions of the sound, and various stages of the decayed, wood. These last are reported in this paper im- mediately after the discussion of the cytolytic enzymes, for the decay is more directly a result of these enzymes. The following are the experiments carried out in the labora- tory to determine the production of ligninase by Lenzites, and incidentally showing the action of cellulase. One gram of fine shavings of the sap-wood of Pinus echinata was placed in each of 3 test-tubes. The shavings had pre- viously been soaked in distilled water for 48 hours to remove as much of the soluble substances as possible, and subse- quently dried. To 1 tube were added 15 ce. of the enzyme dis- persion from the mycelial meal and 15 cc. of distilled water, to the second tube 30 cc. of distilled water, and to the third 15 се. of distilled water and 15 со. of the enzyme dispersion which had been autoclaved up to 10 pounds pressure. To all a few drops of toluol were added. Fifteen days later the liquid was decanted from the shav- ings and filtered. The shavings were boiled in absolute alcohol for 10 minutes, after which the alcohol was decanted off and tested for Czapek's hadromal. With the addition of phlorogluein and hydrochloric acid, the first gave a pink color, while the second and third gave clear, colorless tests. Some of the sections (shavings) were subsequently stained with phloroglucin and hydrochloric acid and others with chloriodid of zinc. Shavings from the first tube were stained a deep red with phloroglucin and hydrochloric acid, but with chloriodid of zinc there was a yellowish color given to all the layers of the walls. Shavings from the second and third tubes gave the same color reaction with the phloroglucin as those from the first tube, but with the zine chloriodid the lamella next to the lumen was stained a light purple, while the outer lamellae of the wall took a yellowish color. These reactions show conclusively that hadromal is split off in the presence of an enzyme preparation from the mycelium of L. saepiaria; [VoL. 3 470 ANNALS OF THE MISSOURI BOTANICAL GARDEN that the same enzyme preparation is capable of hydrolyzing the free cellulose of the inner lamellae and leaving only such substances as will give a yellow reaction with chloriodid of zinc; and that the active substance in the preparation is thermolabile. The aqueous solution, just as it was filtered from the shav- ings, reduced Fehling’s solution; that from the second and third tubes reduced Fehling’s somewhat, but in comparison not so strongly as that from the first. Thus, quantitative de- terminations were made to decide whether the reducing sub- stances were due to enzyme action or possibly to the presence of other reducing substances already in the wood, as, for in- stance, tannins. То this end 10 сс. of the aqueous solution from each were placed in Erlenmeyer flasks of 125 ee. capacity. A fourth determination was made as a control on the Fehling’s solu- tion. The determinations of reducing sugars were made as glucose by the Shaffer method (74). Table уш gives Ше results obtained: TABLE VIII REDUCING SUGARS DUE TO ENZYME ACTION OF LENZITES SAEPIARIA ON PINE SHAVINGS IN 15 DAYS Tube . Reducing Net number Experiment substances bove as glucose | controls mg. mg. 1 1 gm. pine + Va cc. enzyme dispersion + 15 cc. distilled water.............................. 10.606 8.973 2 1 gm. pine + 30 cc. distilled water.............. 1.492 | raps 3 1 gm.pine 4- 15 cc. раста chu (autoclaved) am E. IE MM ROMPE D 1.698. | рай 4 Fehling’s aps сесин e sins x TENERE T Negligible | ........ The results obtained here show that these reducing sub- stances are due to enzyme action, and that they must be sugars, probably glucose and other monosaccharides. The sources of this glucose may be various. There is probably some tannin, although slight, in sap-wood. Besides this there is the hydrolysis of cellulose to reducing sugars, as demon- strated by the experiments reported later on cellulase. Then 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 471 there are probably some starches of the medullary rays hydro- lyzed by diastase, and coniferin hydrolyzed by emulsin, yield- ing glucose and coniferyl alcohol. To amplify the results obtained above, a few pieces of a pine block which had been in pure culture of L. saepiaria for a period of 6 months were extracted with absolute alcohol for 10 minutes, and the amber-colored extract yielded a deep red, with the addition of phloroglucin and hydrochloric acid. A quantity of wood from a railroad tie decayed by L. saepiaria was extracted with alcohol for 10 hours in a reflux condenser. The filtered alcoholic extract obtained in this way was of a deep amber color. When a small portion was diluted to 2 volumes with alcohol and tested with phloro- glucin and hydrochloric acid, it gave a deep sherry-red. When the alcoholic extract was evaporated to dryness a hard amber- like residue remained. This breaks with a conchoidal frac- ture, and seems to be identical with Czapek’s hadromal. These results on a typical brown rot are the same as those found by Czapek (799*) for the brown rot produced by Meru- lius lacrymans; that is, there is a substance, which gives Ше lignin reaction, set free by enzyme action. In the case of Lenzites decay, however, the cellulose disappears as rapidly as it is set free, and in this respect the action is more rapid than in Merulius. The same type of experiment was repeated, using the enzyme dispersion prepared from the sporophoral meal, but there were no results worthy of mention, other than the fact that the shavings in all cultures gave the same tests as were obtained in the water control in the above series. Quantita- tive determinations of reducing sugars were not considered worth while, as the reduction of Fehling’s solution was so slight that no visible copper oxide was thrown down. Cellulase.—True or normal cellulose forms the groundwork of the plant cell wall in most instances. It is a complex carbo- hydrate of the formula (CeéH1i005)n. It is distinguished by its great resistance to hydrolysis and its insolubility in most chemical solutions. To the researches of Cross and Bevan (7014, ’06, 712), we are indebted for a great deal of our knowl- [VoL. 3 472 ANNALS OF THE MISSOURI BOTANICAL GARDEN edge of the celluloses. A full review of the chemistry of cellu- lose is given by Schwalbe (’11) in a very comprehensive way. It suffices to say here that the decomposition products of cellu- lose are mono- and disaccharides, and the decomposition may be brought about more or less readily, according to the com- plexity of the cellulose molecules, by the action of acids or alkalis and by the hydrolysis due to enzyme action. An adequate review of the literature concerning the róle of microorganisms and filamentous fungi in the fermentation of cellulose, especially in the soil, may be had by recourse to the papers by Kellerman and McBeth (’12) and McBeth and Seales (713). Much of the literature on early experiments with parasitic fungi and their ability to pierce the cell mem- brane is reviewed in the above-mentioned papers, as well as by Cooley ('14). Space will permit only a brief review of some of the more important later papers dealing more closely with the destruction of the true cellulose of wood fibre after it is set free from the lignocellulose. Czapek (799*) found in the decay produced by Merulius lacrymans that the cellulose disappeared from the cell walls, and he concluded a priori that a cellulose-hydrolyzing enzyme was excreted by the fungus, although he was unable to demon- strate it experimentally. Ward ('97) observed that in the progress of wood destruction due to Stereum hirsutum the action proceeds from the lumen outward. The sound wood (Aesculus) gives no cellulose test, but the first signs of dissolution are the swelling of the layers next to the lumen and the separation of the lumen from the layers next to the middle lamellae. These swollen layers give the test for cellu- lose, and as they disappear the next layer becomes delignified, gives the cellulose test, and finally disappears. The middle lamellae remain untouched. Ward did not attempt to isolate the enzyme which hydrolyzed the cellulose. According to Buller (’05), wood is rotted by Lentinus lepideus in much the same manner as by Merulius and Stereum. It shrinks and cracks on drying, and is then very brittle and friable. The free cellulose is removed by the fungus. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 473 It seems from experiments carried out by Kohnstamm (201) that the juice of Merulius lacrymans, expressed according to Buchner’s process, gave evidence of the existence of a true cellulase. He found, that after 50 hours of action of the ex- pressed juice on leaves of Elodea, there was a corrosion in the form of fine lines extending out from definite spots in the walls. These streaks soon became thin, and the walls appeared to be obliquely marked with alternate light and darker lines. He mentions that this corrosion in these definite lines, which are always in the same direction, is influenced apparently by the micellar structure of the cell walls. Buller ('06) attempted to prove the presence of cellulase in the sporophores of Polyporus squamosus in the following way: Thin sections of barley grains, which had been cleared of starch by means of the action of saliva, were placed in the fungous extract. No indication of cellulase was obtained, but owing to the disappearance of the cellulose from diseased wood, he assumed that the vegetative part of the fungus pro- duced abundant cellulase. In Falck’s (209, p. 156) discussion of the destruction of fir, pine, and spruce wood by means of Lenzites, he says that in the beginning of the destructive stages the lignin reactions are decreased, and in the last stages they have almost com- pletely disappeared, but that the reactions for cellulose are negative in all stages of decay. On the other hand, in dealing with the same fungus Spaulding (711) says that **phloro- glucin and hydrochlorie acid give a bright red in the rotted балақ ..... Chloriodid of zine gives a blue color only in part of the tissues in early stages of the disease, but in later ones it gives blue throughout.” This would seem to indicate that the fungus had disorganized the lignocellulose, but had left the free cellulose and most of the hadromal. Reed (713) grew Glomerella rufomaculans upon a nutrient solution containing strips of filter paper. There was con- siderably more growth in the flasks containing cellulose than in the controls. At the end of 2 months there was somewhat more than 3 times as much dry fungous matter in the regular as in the controls. The solution gave no tests for reducing [Vor. 3 474 ANNALS OF THE MISSOURI BOTANICAL GARDEN sugars, which were probably utilized by the fungus as fast as they were split off. I Wolf ('16) made poured plates of cellulose agar which he inoculated with species of Pseudomonas, Phoma, Gloeo- sporium, and Fusarium. He says: “There was no evidence of the production of cellulase except by Phoma.’’ Samples of normal tissues and of tissues diseased by Phoma socia Wolf were tested for cellulose by employing Schweitzer’s reagent, and Wolf observes that ‘‘there is a slight but significant de- crease in the amount of cellulose found in diseased tissues."' A considerable number of determinations consistently showed that “the lesser amount of cellulose was invariably found in the diseased tissue.’’ Further experiments were instituted, since it was clear from microchemical tests and from the above experiments on lig- ninase that cellulose of attacked wood disappears. In order to test out the action of the enzyme preparations on normal cellulose, pure cellulose from two sources was prepared. One was prepared from filter paper in the way described by McBeth and Scales (713) and later by Cooley (714). Fifteen grams of filter paper were dissolved in Schweitzer’s reagent and precipitated with dilute hydrochloric acid. After wash- ing thoroughly with dilute acid to get rid of all of the copper and then with distilled water to get rid of all of the chlorine, a very flocculent cellulose precipitate was obtained. The water was filtered off with a Buchner funnel until the cellulose suspension was concentrated to about 500 cc. This was trans- ferred to a liter flask which was plugged and sterilized. Another type of pure cellulose was made from pine wood. A quantity of fine pine shavings were treated with a cold solu- tion composed of 30 grams of potassium chlorate dissolved in 520 се. of nitric acid (sp. gr. 1.1). The container was kept cold for 4 weeks, after which time the cellulose was washed and then precipitated from Schweitzer’s reagent, as in the above case. It was sterilized and kept for future use, as was the filter paper cellulose. Several types of cellulose agar were prepared, using both the filter paper cellulose and the pine wood cellulose. Some 1916] ZELLER—TPHYSIOLOGY OF LENZITES SAEPIARIA 475 standard nutrient solutions for the artificial culture of fungi were used as a basis for these, such as: (1) Richards’ (797) solution, substituting 100 ec. of the 2 cellulose suspensions for cane sugar and adding 2 grams of agar. БЕС ec MO E 10 gram ОООО г... 05 gram E EE 0.25 gram Ss PR EE 0.002 gram Cellulose suspension..... 100 ec. E E EE 2 grams (2) Cooley’s solution “A,” just as given by Cooley (714, p. 306). (3) Reed's solution, as given by Reed (713, p. 69), with the exception of using one-half as much distilled water to- gether with 500 ec. of cellulose suspension and 2 per cent agar. On any of these agars L. saepiaria grows very slowly and without producing much of a mat. Wherever cellulose hydrolysis could be seen it was very, very slight, and this was only in cases where the pine wood cellulose was used. Sinee these experiments gave such meagre results 2 per cent agars were made, using dilute extracts of carrot, turnip, and potato as bases and using cellulose suspensions in ap- proximately the same concentrations as in the above. In this series of experiments the cloudiness of the agar due to cellu- lose suspension was cleared up noticeably in one case only. This was where carrot-pine-wood-cellulose agar was used. The agar was tubed and sterilized in test-tubes of 13 mm. diameter. The agar was not slanted, and after inoculation the tubes were kept in a damp chamber so that the water con- tent of the agar would remain the same throughout incuba- tion. These tubes were kept at a temperature of 32°C. for 4 weeks. After this length of incubation the agar had cleared to an average depth of 9 mm. in the inoculated tubes where carrot-pine-wood-cellulose agar was used. The uninoculated tubes were still uniformly cloudy. There was no hydrolysis in the tubes where carrot juice was not used. The most sig- nificant fact brought out in this series of experiments is that [VoL. 3 476 ANNALS OF THE MISSOURI BOTANICAL GARDEN the pine wood cellulose is hydrolyzed by the cellulase excreted by L. saepiaria, while the filter paper cellulose remained untouched. Experiments with the enzyme dispersion were set up accord- ing to the scheme as outlined in table 1x. All of the experi- ments were set up in duplicate with both sporophoral and mycelial dispersions. Toluol was used as an antiseptic, and qualitative results were taken after 4 weeks. TABLE IX d EXPERIMENTS SHOWING THE ACTION OF MYCELIAL AND SPO- ORAL ENZYME DISPERSIONS ON F pilo е CELLULOSE D PINE WOOD CELLULO Reduction of Fehling's solution Experiment Mycelium Sporophore 10 cc. paper cellulose + 2 cc. dispersion. ......... + + + 10 cc. pine lose + 2 dispersion............ + + + + + + 10 cc. pine cellulose + 2 cc. dispersion (autoclaved) + — 10 cc. paper cellulose +2 cc. dispersion EE d)| + -- 10 сс. paper cellulose + 2 сс. distilled water........ = Sp 10 cc. pine cellulose +- 2 cc. distilled cn? Kee, — - Since such definite results were obtained with the mycelial enzyme dispersion in the foregoing experiments, it was de- termined to make quantitative comparisons of the activity of cellulase in the sporophoral and mycelial tissues. To this end a carrot extract was made as described above under ‘‘inver- tase,’’ and to this extract was added pine wood cellulose. This nutrient solution was placed in Erlenmeyer flasks, which were plugged, sterilized, and inoculated as described for invertase and diastase. The mats of mycelium formed after 2 weeks were removed from the flasks, dried, and ground to a fine powder which was used in the following way: To 50 cc. of the pine-cellulose suspension were added 2 grams of the mycelial powder with toluol as an antiseptic. Some of the mycelial powder was autoclaved, and controls on the above were pre- pared with this autoclaved powder as well as with distilled water alone. These experiments were set up in duplicate. A parallel series was prepared using the sporophoral tissue powder. After incubating these enzyme cultures at about 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 477 28°C. for 4 weeks the sugar content of the substrates was de- termined as glucose by the Shaffer method already mentioned. The results of these determinations are reported in table x. In the light of these results, there is no doubt that cellulase is present in the mycelium of Lenzites and that its activity can be measured quantitatively. The little activity shown in the fruiting bodies is probably due to the small amount of superficial mycelium which was removed from the substrate TABLE X QUANTITATIVE DETERMINATIONS OF SUGAR PRODUCED BY THE CELLULASE OF LENZITES SAEPIARIA Reducing sugar as glu- N Amount of tissue powder in cose in 50 cc. of sub- 50 cc. of pine wood-cellulose strate after 28 days t glucose above controls Mycelium | Sporophore | Mycelium | Sporophore mg. mg. mg. mg. 3 Ием pow WEE 6.479 0.99 3.271 0.28 n be ER, tri Oe AXES... расата е alone....... Negliibie] ....- ЕЕЕ ке: when the sporophores were collected. It is almost impossible to get purely fruiting tissue without some such closely con- nected vegetative tissue, and it is also impossible to say whether or not the enzymes diffuse from the adjacent myce- lium to the base of the fruiting bodies. A series of quantitative cellulose determinations has been made to establish further the cellulase activity in L. saepiaria. It was thought that possibly there might be established some relation between the percentage in reduction in weight due to deeay by the fungus and in the loss in cellulose due to the cellulase, or some relation between either of these and the specifie weight of the substrate. But three de- terminations with their eontrols have been made. Blocks of yellow pine, approximately 1 12 inches, were dried to constant weight and weighed, and the volumes were taken by immersion in mereury. From these figures the specific weight was computed. These blocks were sterilized in jars plugged with cotton, inoculated with L. saepiaria, and incubated under [Vor. 3 478 ANNALS OF THE MISSOURI BOTANICAL GARDEN favorable moisture and temperature conditions for one year. After this period they were dried, and the percentage of re- duction in weight due to fungous decay was determined. Con- trol blocks of the same specific weight and from the same samples were kept in a sound condition. To make the cellulose determinations the blocks from the cultures and the controls were planed into fine shavings. All of the shavings from each sample were placed in a 250-сс. TABLE XI "n DETERMINATIONS OF CELLULOSE FROM BLOCKS OF PINE WOOD, OTH SOUND, AND DECAYED BY LENZITES SAEPIARIA g: ча ч 9 әс» So 2 o e 25 8 es Bs*:|zsE ОП 28 | 8.8 Se? FE; 3 $3 | bes | Boo] зе] BS | 252) 25 He} 8 ne) og 355 RES 28 Ié glans gm. gm. gm. OS | Decayed | .424 | 13.373 | 6.973 | 47.86 | 0.9563 | 13.714 | 43.057 05 Sound See FE ek Lu asl 2.0592 | 24.0848 Kee F5 Decayed | .419 | 12.580 | 8.494 | 32.48 | 1.2952 | 15.248 | 38.187 F5 Sound .419 Ito We a ere IAN D 2.3066 | 24.668 |........ A 24 Decayed | .547 | 12.418 | 5.84 52.97 | 0.1943 | 3.327 | 75.654 A 24 Sound .547 LUE E ceu 0.9703 | 13.666 |........ Erlenmeyer flask containing about 125 ee. of the solution of potassium chlorate in nitrie acid mentioned above. These were placed in an ice chest for 3 weeks, after which time the contents of each flask was diluted to about 3 liters. The di- luted liquid was thus weak enough not to attack a filter paper while filtering. The liquid was filtered off through a Buchner funnel containing a tared filter paper. The cellulose thus ob- tained was repeatedly washed with hot distilled water until it was of a pure white color and gave a deep blue reaction with zinc chloriodid. In all cases the yield of cellulose was so clear of foreign material that it was deemed unnecessary to precipi- tate from Sehweitzer's reagent. After drying and weighing, the percentage loss in cellulose due to the action of the fungus was determined. The different factors in this experiment are tabulated in table хі. In these 3 determinations there seems to be no definite re- lation between the percentage loss of cellulose and the percent- 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 479 age reduction in weight due to fungous decay. This may be due to different proportions of lignifieation in the different samples, or more likely, since other substances like coniferin, hadromal, and possibly vanillin are utilized by the fungus, the total reduetion would not necessarily bear any defmite rela- tion to the reduction of any one of the complex. Helbig (711) has made similar cellulose determinations on wood which had been altered by Thelephora perdix. This fungus produces a white rot, and Helbig found that with the advance of decay the percentage of cellulose increases perceptibly. Hemicellulase.—The hemicelluloses differ from the true celluloses in that they are more easily hydrolyzed, are readily dissolved in hot dilute acids, and sometimes give a blue color with iodine. Their chemical compositions are determined by the products of their hydrolysis. They may yield dextrose, mannose, galactose, or mixtures of these, and at times xylose or arabinose. According to these decomposition products, they are differentiated into dextrans, mannans, galactans, mannogalactans, ete. Very frequently hemicelluloses are de- posited upon, or as a part of, cell walls, and here play the róle of reserve foods, especially in seeds. Newcombe (799) determined that the cellulose-hydrolyzing enzyme is distinct from other earbohydrases. Schellenberg (208), through experiments on numerous fungi, also proved that pure eultures grown on substrates containing hemicellu- loses and true celluloses would hydrolyze hemicelluloses but not the true celluloses. He also shows that the moulds act selectively toward the hemicelluloses from various sources. He thus differentiates between the different hemicellulases which act on various hemicelluloses, and the cellulase which hydrolyzes the true cellulose. To ascertain whether the enzyme dispersions from mycelium and sporophores are active as hemicellulose-hydrolyzers, the endosperm of the date seed (Phoenix dactylifera) was used as a substrate. Date seeds were scraped to remove the outer coats, and then were thoroughly washed with sand and soap to remove all reducing sugars possible. The seeds were then rinsed in dis- [Vor. 3 480 ANNALS OF THE MISSOURI BOTANICAL GARDEN tilled water, cracked, and the embryos removed. The hemi- cellulose thus prepared was autoclaved in distilled water at 12 pounds pressure for 20 minutes to kill all enzymes present. The water was again decanted off, and the endosperms rinsed and allowed to remain in distilled water with toluol to pre- serve for future use. Van Tieghem cells were prepared, and very thin slices of hemicellulose were suspended in hanging drops of enzyme dis- persion, as follows: (1) Eight cells with hanging drops of mycelial dispersion. (2) Eight cells with hanging drops of sporophoral disper- sion. (3) Four cells with hanging drops of autoclaved mycelial dispersion. (4) Four cells with hanging drops of autoclaved sporo- phoral dispersion. (5) Four cells with hanging drops of distilled water. In the bottom of each cell was placed enough of the solu- tion of the same vapor tension as the hanging drop, so that evaporation of the drops was prevented. A drop of chloro- form was added to each cell as an antiseptic. These cells were examined from time to time, but no sign of the erosion of the hemicellulose was noticed until after 25 days. There was a slight indication of erosion in five of the drops of mycelial dispersion. The other three were contaminated with bacteria and showed slight erosion. There was no erosion in the con- trols except in one contaminated with bacteria. After 40 days three of the five drops of mycelial dispersion observed 2 weeks before were still perfectly aseptie, and two had dried down. The pieces of hemicellulose in the three cells were strongly eroded. In places only a granular substance was left. When the cover glasses were removed from these three cells there was a strong odor of chloroform still remaining in each. That they were perfectly aseptie was proven by removing from the drop what remained of the slice of hemicellulose, drying down the hanging drops on the cover slips, and flaming and staining the smear with gentian violet. There were no bac- teria or fungi present. There was no erosion in any of the 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 481 cells eontaining the enzyme preparation from the sporophores nor in the cells containing the mycelial dispersion which had been autoclaved. These results go to show that there is hemi- cellulase in the mycelium of L. saepiaria but not in the tissues of the fructifications. Another experiment was conducted as follows: Four test- tubes each were prepared with mycelial and sporophoral dis- persions in the following manner: (1) 0.5 gm. hemicellulose+10 се. enzyme preparation--to- luol. (2) 0.5 gm. hemicellulose--10 се. enzyme preparation --to- ol. lu (3) 0.5 gm. hemicellulose+10 сс. enzyme preparation (autoclaved) +toluol. (4) 0.5 gm. hemicellulose--10 ce. distilled water--toluol. These were incubated at 25-30°C. for 25 days, after which the liquid was filtered off and 5 се. from each were tested with Fehling's solution for reducing sugars. Numbers 1 and 2 showed slight reduction of copper produced by the mycelial dispersion but not by that from the sporophores. In the con- trols no copper oxide could be detected. From these results it is certain that the mycelium contains the enzyme, hemicellulase, capable of hydrolyzing the hemi- cellulose of the endosperm of Phoenix dactylifera. This hemi- cellulose is a paragalactan, which, on hydrolysis, yields a mix- ture of galactose and arabinose, both of which reduce F'ehling's solution. Pectase and pectinase.—Closely allied with cellulose is a group of substances called pectic bodies. Pectose is the name given to the parent substance of bodies, such as pectin, pectic acid, ete. Many fruits, such as apples, gooseberries, currants, cranberries, and fleshy roots—such as carrots—contain a sub- stance soluble in water but gelatinizing in aleohol. This sub- stance which causes the juice of fruits to ‘‘jell’’ is known as pectin. A solution of pectin gelatinizes on standing, probably due to the action of the enzyme, pectase, contained in the fruit juice. [Vor. 3 482 ANNALS OF THE MISSOURI BOTANICAL GARDEN Mangin (792, '93) investigated the pectose group of sub- stances and divided them into two groups: first, neutral bodies varying in their solubility in water from pectose, which is insoluble and closely resembles cellulose, to pectin, which is soluble but readily forms a jelly; second, acid bodies, chiefly pectic acid, the latter occurring as calcium pectate, forming the middle lamellae of plant tissues. The enzyme which is capable of hydrolyzing the pectic bodies is generally termed pectinase, while the one causing coagulation is pectase. It is a well-known phenomenon in certain types of decay that the middle lamellae disappear. As early as 1886, de Bary (286) observed that the mycelium of Peziza sclerotiorum was capable of penetrating cell walls and gelatinizing them. The juice of the sclerotia of this fungus had the power to dissolve the middle lamellae and gelatinize the inner layers of the cell walls of turnips and carrots. The enzyme preparation precipitated from the juice by means of alcohol affected the cell walls in the same way. Ward (’88) observed the macerating action of the Botrytis causing the lily disease. These observations were made on sections of the leaves, petioles, and ovary of the lily. The middle lamellae underwent dissolution in a few hours when placed in aqueous extracts of the fungus. Since Jones (205) and Cooley (714) have so amply reviewed the earlier work in this field, it will not be diseussed further in this paper. Cooley (714) shows that in tubes containing pectin a coagulum was produced by Sclerotinia cinerea, thus showing the exere- tion of pectase by this fungus, which, nevertheless, shows no particular affinity for the middle lamellae. On the other hand, Brown's (715) work with Botrytis cinerea shows this fungus to possess the power of dissolving the middle lamellae. The enzyme extract prepared from very young mycelia brought about a very rapid disintegration in the tissues of potato, turnip, beet, apple, etc. Discs of these tissues were dis- organized in from 15 to 90 minutes. The death of the cells did not take place until some time after they had been separated by the solution of the middle lamellae. The activity of the extract was destroyed by heat. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 483 No work on wood-destroying forms has demonstrated the presence of an enzyme capable of disorganizing the middle lamellae, although many of the decays show maceration. Spaulding (711) says that in the last stages of decay рго- duced by Lenzites saepiaria the middle lamellae have disap- peared. Since it was found in microchemical observations that the middle lamellae of the wood decayed by L. saepiaria were dis- solved out, it was attempted to demonstrate pectase and pecti- hase experimentally. Pectin was prepared in the usual way by the action of alcohol on the juice expressed from cran- berries. Pectase from the carrot coagulated this pectin, but only negative results were obtained with the enzyme disper- sions from L. saepiaria. Further experiments were con- ducted to determine the macerating power of the dispersions on various tissues. Slices of carrot, potato, and beet were cut to a uniform thickness. From these slices dises were cut by means of a cork-borer, and similar discs were also prepared from very young tobacco leaves. As a source of enzymes, the mycelial powder prepared from mycelium grown on carrot juice was used. Two grams of this powder were soaked in 50 сс. of distilled water for 5 hours, after which the liquid was deeanted off. 'The four kinds of dises mentioned above were placed in portions of this liquid in closed stenders and a few drops of toluol added to each. Dises were also kept in dis- tilled water as controls. Observations were made after 18 hours. Carrot discs in the mycelial extract had lost in coher- ence in comparison with those in the distilled water. When pulled apart the latter were torn as much across the cells as following the cell walls, while in the former the separation fol- lowed the line of the cell walls. Beet dises showed no macera- tion whatever after 18 hours. Potato dises showed a more marked maceration than the carrot. The potato had become very flaccid in the extract. The discs of tobacco leaves showed no loss of coherence. After 42 hours the carrot discs and potato discs had lost all coherence, and the cells were easily pressed apart under a cover glass. In the controls in [Vor. 3 484 ANNALS OF THE MISSOURI BOTANICAL GARDEN distilled water there was no diminution of coherence of the tissues. Thus, the presence of pectinase may be demonstrated in the mycelium of L. saepiaria, while there are no indications of the presence of pectase. The effect of the cyto-hydrolyzing enzymes as demonstrated by microchemical observations on the sound and decayed wood.—Pine wood is composed of tracheids with one row of bordered pits in the radial walls. The annual rings are usually well differentiated into spring and summer wood, especially in the heart. Resin ducts occur among the tracheids, extending longitudinally, as well as radially, in the medullary rays. The resin ducts are surrounded with wood parenchyma. The sap-wood is lighter in color than the heart, probably due to oxidation, since the lumen of the heart tracheids are well aérated, as well as to the presence of tan- noid bodies which become darker with continued exposure to air. The sap-wood, of course, is not so thoroughly lignified as the heart-wood, and often the inner lamella gives the cellu- lose test with zine chloriodid. The wood that has been attacked by L. saepiaria is darker in color than the sound wood; it is also very brittle and when crushed between the fingers breaks into a fine powder. It is evident that marked changes take place in the wood due to the action of enzymes produced by the fungus. To determine what some of these changes are and something about their sequence, I resorted to microchemical tests. To this end sec- tions of sound wood and wood in various stages of decay were examined. Free-hand sections were made longitudinally, but in the later stages of decay it is impossible to cut transverse sections because of the brittle character of the tissues. Small pieces, carefully cut down to 0.5-се. cubes, were imbedded in celloidin, and from these then the free-hand, transverse sec- tions were cut. The sound sap-wood gave the following tests: (1) An alcoholic solution of phloroglucin with an addi- tion of hydrochloric acid gave a deep red in the middle lamella, dark red in the secondary, and pink in the tertiary lamella. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 485 (2) After soaking sections in iodine and then treating with 65 per cent sulphuric acid, the secondary lamella was a yellow to brown color, with a purple lining in the early spring wood. (3) Chloriodid of zine gave a brown coloration both in the spring and summer wood, with a slight blue tint lining the tracheids in the sap-wood. (4) Aniline sulphate produced a bright yellow which in- creased in intensity from spring to late summer wood. (5) After treating with potassium hydroxide for some time and then applying the cellulose tests the following results were obtained: (a) After continued action of iodine followed by the addition of sulphuric acid, the inner or tertiary lamella was swollen and shrunken away from the secondary, the former assuming a purple color. (b) Chloriodid of zine colored the swollen tertiary lamella blue, while the main part of the wall was brown. (6) Resorcin and sulphuric acid gave a violet to blue re- action in the lignified walls. When these same tests were applied to the decayed wood some difficulty was found, especially where the tests yield yellow or brown, because the tissues were decidedly brown in the last stages of decay. If a transverse section is made through a decayed portion of wood so as to include a part of the sound, normal wood, the progressive stages of decay may be followed by applying the above stains. When phloroglucin and hydrochlorie acid are applied to such a section, it is noticed that chemical changes have pre- ceded any visible or microscopical changes in structure. In the sound wood the secondary and tertiary lamellae are stained a dark red, while the middle lamella is a still darker red. A little nearer the edge of the decayed region the red has changed to a maroon or brownish red in the tertiary lamella. As we proceed nearer to the decayed portion this maroon increases until the secondary lamella is all brownish red, and the middle lamella only remains the brighter red with [VoL. 3 486 ANNALS OF THE MISSOURI BOTANICAL GARDEN this stain. In radial sections it may be noticed that this maroon discoloration starts from the bordered pits, especially if they are perforated with fungous hyphae. The hyphae seek the bordered pits, these apparently serving as the only places where the hyphae pass from one lumen to another. In the tangential section the discoloration in the earliest stages occurs only in the neighborhood of the medullary rays. While this discoloration is taking place the tertiary lamella first contracts and then practically disappears. The second- ary lamella likewise shrinks as it takes on this brownish red color. This must be due to the gradual hydrolysis of cellu- lose and probably the simultaneous hydrolysis of coniferin through the action of emulsin. The secondary lamella shrinks, but in the last stages of decay there are still brown, fragile remains of this layer, together, undoubtedly, with infiltrated by-products from the decayed middle and tertiary lamellae. The middle lamella seems to disappear almost simultaneously with the decay of the secondary. With chloriodid of zine the decayed wood gives the same test as in the sound wood, a brown color. This is true in all stages of decay. There is no indication of free cellulose at any time during decay. Partially delignified sections, cut from the same surface as the sections on which the above lignin tests were made, were treated with a 5 per cent potas- sium hydroxide for some time. On the addition of chloriodid of zinc the lamellae yielded a purplish blue reaction, where only partially delignified by the fungus; but where decay was complete a brown color was obtained. These results show that the first step is the splitting of lignin, and simultaneously with this there is a complete hydrolysis of the cellulose as fast as it is set free. Undoubtedly some of the substances giving the lignin reaction are also used up. Other lignin tests gave similar results. The action of ani- line sulphate was marked. The sound wood was colored yel- low, and as the diseased region was approached the color be- came browner, although the yellow element did not seem to be lost entirely. Indications are that in the decayed wood some of the substance that gave the lignin reaction still — 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 487 remained. This was easily extracted and is what Czapek has called **hadromal." The iodine-sulphurie acid test for cellu- lose corresponds well with what we found with zine chloriodid — a light brown color. Our microchemical tests applied to the decayed wood sub- stantiate in the main the results obtained with the enzyme dispersions and other enzyme preparations, i.e., that cellulase and ligninase are secreted by L. saepiaria. Pectinase is both demonstrable in vitro and in nature, the pectinase of the middle lamella disappearing with the action of the fungus. A point of further interest is the composition of the brown substance left after the complete decay of the tracheids. This is a brittle substance which is easily crushed into a fine brown powder. A quantity of this brown material secured from the deeayed eavities of an old railroad tie was ground as finely as possible in a mill, and to this powder was added a dilute alkali. After soaking for 2 days the alkali was filtered off, and by adding acid to the filtrate a flocculent precipitate was thrown down. When dried down this precipitate shrinks and cracks. It is insoluble in chloroform, alcohol, ether, acetone, and petroleum ether, but is readily redissolved in alkali and may be reprecipitated with acid. This substance partakes of the nature of ‘‘humus’’ compounds. The remainder of the brown powder is much like ‘‘peat.’’ In the disease of Taxodium distichum known as “реску” cypress, von Schrenk (700”) found a similar substance which he called a humus compound. In the case of Taxodiwm the humus compounds are in a liquid form, and thus are deposited in the tracheids where the mass dries and cracks, ‘‘looking much like mud which has dried in the sun." The humus liquid infiltrates into the sound wood immediately surround- ing the decayed spots and darkens the wood in the decayed regions. Undoubtedly, the humus compounds found in wood decayed by Lenzites and by the fungus causing the peckiness of eypress are a direct result of the activity of the enzymes concerned in the decay. It was mentioned above that in the process of delignification, etc., the shell of the tracheids remaining after the last stages [VoL. 3 488 ANNALS OF THE MISSOURI BOTANICAL GARDEN of decay shrinks and cracks. The cracks in the walls always follow the same general direction and seem to begin with the bordered pits. The slits are spiral in form and pass obliquely across the pits from left to right upwards. By changing the focus these lines are from right to left upwards on the farther wall of the tracheid. Von Schrenk ('00") also observed this in the tracheids of Taxodium distichum infected with **pecki- ness.’’ In the erosion of the cell walls of Elodea leaves, due to the action of an enzyme preparation from Merulius lacry- mans, Kohnstamm (’01) noticed that the action took place in definite lines. He is inclined to believe that this is due to the micellar structure of the cell walls. In general, the Lenzites decay is of the same type as that produced by Merulius lacrymans and reported on by Czapek (’99), but the hydrolysis of the cellulose is much more rapid in the former. In contrast to this type we have the other ex- treme represented by the pin rot of pine due to Trametes Pini, the white rot of cedar due to Polyporus juniperinus, and the rot of oak produced by Thelephora perdix. In this case, as mentioned before, all substances are used by the fungus with the exception of cellulose which is left as a pure white by-product. INULINASE Inulinase was discovered by Green (’88) in the Jerusalem artichoke (Helianthus tuberosus). It was first demonstrated in fungi by Bourquelot (’93) who found it in Aspergillus niger, but later with Hérissey (’95) did not find it in the sporo- phores of Polyporus sulphureus. Dean (’03) verified Bour- quelot’s work with Aspergillus niger and Penicillium glaucum and found inulinase to be an intracellular enzyme. Dox (710) reported that Penicillium Camemberti had slight action on inulin unless cultivated on a substrate containing inulin as the source of carbon. In this case inulinase was produced more abundantly. In our experiments with Lenzites the enzyme preparations from the mycelium and the sporophores were used. To 10 cc. of a 1 per cent solution of inulin 2 cc. of the enzyme dispersion were added and toluol used as an antiseptic. There was a 1916] ZELLER— PHYSIOLOGY OF LENZITES SAEPIARIA 489 marked reduetion of copper oxide from Fehling's solution after 2 days of incubation at 25-30°C., while the boiled con- trols and water controls showed no reduction. The inulinase seemed to be quite active in both the mycelium and sporo- phores, but no quantitative comparison was made. AMIDASE AND UREASE Since we found positive proof of the presence of tryptic and ereptic ferments in the fungus, the next logical step in sequence was to ascertain whether amidases were present, for protein digestion normally proceeds further than to the pep- tone stage and results in the amino acids, which, digested by amidases, yield ammonia and hydroxy-acids. In the lower fungi these desamidizing enzymes have been found by many workers. Butkewitsch (’03) found by growing cultures of Aspergillus, Penicillium, and certain species of Mucor on liquid media, containing proteins that ammonia is liberated; and in the following year Shibata (204) found in a ‘‘Dauerpraparat’’ of the mycelium of Aspergillus niger an enzyme which split ammonia from different nitrogen-contain- ing substances, while Pringsheim (’08) found the same enzyme present in ‘‘Acetonedauerhefe.’’ Dox (70) found that the enzyme preparation from Aspergillus niger and Penicillium Camemberti showed the power to split ammonia from aspar- agin and urea. Reed (713) found very similar results by the action of the enzyme powder from Glomerella rufomaculans on asparagin and alanin. The only record of urease from one of the higher fungi was that reported by Kikkoji (’07) in Cor- tinellus edodes. In the experiments reported in the following tables the enzyme dispersions from both the mycelium and sporophores and also the fungous powder from both tissues were used. As substrates 50 cc. of 1 per cent solutions of asparagin, acet- amid, and urea were used. Ten cubic-centimeter portions of the enzyme dispersions were used in some cases, while an equivalent weight of the fungous meal was used where the enzymes were not extracted and precipitated. Autoclaved controls, as well as the substrates alone, were set up for each 490 ANNALS OF THE MISSOURI BOTANICAL GARDEN [VoL. 3 series, and toluol was used as an antiseptic in all cases. Each experiment was set up in a gas-washing bottle, which was fitted with rubber tubes securely stoppered. These washing bottles were incubated at 25-30°C. for 20 days, when the am- monia was determined by the Folin method. Friedrich’s im- proved gas-washing bottles containing 250 ee. of N/50 HCl were used for the collection of the ammonia. Air was drawn through for 14 hours by means of a Richards’ suction pump, TABLE XII AMIDASE ACTION IN THE MYCELIUM OF LENZITES SAEPIARIA Nitrogen as Substrate Form of enzyme material ammonia set} nitrogen free set free mg. mg. ИТИИ Enzyme «іврегвіоп................. 2.10 0.28 Deae Enzyme dispersion (autoclaved)..... 1,82 |12 ық Ше МЕДИЯ ЕРИ Uy trovi rrr tS 1.13 Leite Lil ET TEE Maale asee a e E 23.55 22.57 Lui E EE Meal Mo Gen E 0.98 ^ | ees Asparagin..... Enzyme «іврегвіоп................. 1.40 0.00 Asparagin..... Enzyme Seege (autoclaved)..... ЫМ ` РРР Ре Asparagin аіопе|............................... DM Lic Asparagin.....| Мег!............................. 0.21 0.07 Asparagin..... Мен ege T: E Ee 0.00-— |.J. БЕ ree ee etam ‚| Enzyme dispersion................. 1.12 0.00 icetamid : me dn (autoclaved)..... 134 аркасы PACOTAIMIC.. 2. 00) МЕД. „а. вовозевввосововововевосе 0.14 0.14 tamid Е Meal (autoclaved). с. 5. ees 0:98 СТОР H tx S o E PETER TERRE TERT E tU EE 0.00 леса TABLE XIII AMIDASE ACTION IN THE SPOROPHORAL TISSUE OF LENZITES SAEPIARIA Nitrogen as Substrate Form of enzyme material ammonia se nitrogen ee set free mg. mg. Dl. РРР Enzyme Фөрегвіоп................. 5.46 — eee Enzyme dispersion (autoclaved)... .. 6.16 | oS Исок S WES ЖӘСЕРІ MMC 646 теве 2. тен Yer Y Ma Sine SCR EE 18.34 12.18 Wray cele. ven weal саг deg dir € dE Dis 588. — Без киста Asparagin..... Enzyme дізрегвіоп..........:....... 1.89 0.91 Asparagin..... Enzyme ираса (autoclaved). . 0938 — 15: e Asparagin..... JI E 2.24 1.12 Asparagin..... Meal breng ре Ee зул Лет Acetamid...... Enzyme Ерера. ¿71 pap ee 2.03 1.75 Acetamid...... am dëses (autoclaved)..... 028 WER Dum... TL EST 0.28 -- cetamid Veal С a a AME EEEE 0.8%. - (aa 1916] 3 ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 491 then duplicate portions of the collection acid were titrated against N/50 NaOH. Alizarin red was used as an indicator. Tables хп and хит show the results obtained. It is interesting to notice that urease is the only active enzyme in the fungus which produces desamidation, and that this enzyme was not extracted with water. Pringsheim (’08) has shown that the amidases of yeast do not pass out with the water extract but are tenaciously held by the protoplasm. The same may be the case with urease of Lenzites, and since we assume that the enzymes are protein-like bodies it is possible that urease in this case is not like the albumins, which are water-soluble. It may be a globulin, soluble in neutral saline solutions, or glutelin, soluble in weak alkali. It is again of interest here to notice that the enzyme is much more active in the mycelial than in the sporophoral tissue, even without considering the amount of sawdust present in the mycelial meal. Of course, here, as in other plants, it is not possible to connect the presence of urease with any use- ful function in the metabolism of the fungus. With our present knowledge of desamidation in plants the biological importance of urease remains unknown. HIPPURICASE The ability of the lower fungi to bring about the splitting of hippuric acid by enzyme action was first noticed by Shibata (204) in the case of Aspergillus niger. Some years later Dox (709) demonstrated the presence of hippuricase in other species of fungi, namely, Penicillium Camemberti, P. chryso- genum, and P. brevicaule, and he also confirmed Shibata’s results on Aspergillus niger. The enzyme preparation was the ground fungous mycelium after it had been dried by the usual “ Acetondauerhefe’’ method. His method of determin- ing the amount of hippuric acid hydrolyzed is given in a later paper (Dox, "101. After hydrolysis by the enzyme the hip- purie acid solution, which was made up in weak sodium hydroxide, was mixed with the calculated amount of sulphuric acid to combine with the sodium. This was shaken with petroleum ether which dissolves out the benzoic acid. The [VoL. 3 492 ANNALS OF THE MISSOURI BOTANICAL GARDEN latter erystallizes when the petroleum ether is evaporated, and the crystals, after recrystallization from water, melted at 121°C. and had the appearance of benzoic acid. In the con- trols no residue was obtained. The hydrolysis in the case of Penicillium Camemberti was 76 per cent. In 1912 Kossowicz found that enzyme preparations of As- pergillus niger, Mucor Boidin, Phytophthora infestans, Isaria farinosa, Botrytis Bassiana, and Cladosporium herbarum in every case brought about the destruction of hippuric acid. As one of the reaction by-products he identified ammonia, but in no case did he state in how great quantities the ammonia was formed. Reed (713) found that an enzyme powder prepared from Glomerella rufomaculans showed the presence of hip- puricase after incubation for one week. In 1913 Dox and Neidig applied Sórensen's formaldehyde- titration method for the determination of the acidity of amino acids to the hippuric acid hydrolysis. The method is founded on the reaction between formaldehyde and the primary amino group. Hippuric acid has no primary amino group, but after hydrolysis the primary amino group of the glycocoll split off may be neutralized with formaldehyde, leaving the carboxyl unchanged to be titrated against an alkali. In this case Dox and Neidig used N/10 Ba(OH)s. They grew cultures of As- pergillus niger, A. clavatus, A. fumigatus, Penicillium er. pansum, P. Roqueforti, and P. Camemberti on a nutrient solu- tion. The cultures were grown for 1, 2, 3, and 4 weeks, the juice pressed out after grinding the mycelium and used as an enzyme preparation. In all of these fungi hippuricase was found, as well as in taka-diastase (Aspergillus Oryzae). The age of the mycelium had little influence on the production of hippuricase. Titrations for free ammonia showed that in the cultures 3 and 4 weeks old there were slight amounts, if any, of ammonia, and a secondary reaction, or the splitting of glycocoll, is improbable. Our experiments with the meal from the mycelial and sporo- phoral tissues of Lenzites were carried out in the same way as those described by Dox (710). Fifty ecubie-centimeter quan- tities of the substrate were used, and the flasks were ineubated 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 493 at 25°C. for 2 weeks. After this period the benzoic acid was dissolved out with petroleum ether, dried, and then recrystal- lized from water. From 50 ee. of a 1 per cent solution of hip- puric acid were obtained 0.24 grams of benzoic acid, or there was 65 per cent hydrolysis. The melting point was found to be 126°C. Since the melting point of hippuric acid is 187°C. and that of benzoic acid is 121°C., this was regarded as evi- dence that the crystals were benzoic acid. NUCLEASE The presence of nuclease has been demonstrated in various divisions of the plant kingdom. It performs an important function in decomposing the nucleic acids of plant cells, espe- cially in germinating seeds or wherever dissimilation is car- ried on. In the fungi nuclease has been found both in the lower and higher forms. Iwanoff (’03) studied the effect of cultures and enzyme extracts of Aspergillus niger and Peni- сит glaucum on nucleic acid, and observed that both species produeed the purin bases and phosphorie acid in the sub- strate. He claims that the nuclease is distinct from the pro- teolytic enzymes, for his enzyme extract would not liquefy gelatin. Dox (710) found that the nuclease of Penicillium Сатетеги is formed irrespective of the presence of nucleic acid in the culture medium. Kikkoji (’07) expressed the juice from an agaric, Cortinellus edodes, and 25 ee. of this in 150 ee, of a 2% per cent solution of the sodium salt of nucleic acid produced 28.7 mg. of phosphorus pentoxide in 5 days’ diges- tion. He considered this action due to nuclease, the juice being thermolabile. In my work with Lenzites заетата а 1 per cent solution of phyto-nuclein from yeast was prepared by dissolving the phyto-nuclein in N/20 sodium hydroxide and then neutraliz- ing. To 25 ce. of this solution 5 cc. of the enzyme dispersion were added with toluol as an antiseptic. Controls were set up by adding 5 ce. of autoclaved dispersion to the nuclein solution, and water controls made by adding 5 cc. of distilled water to the nuclein. This plan was followed both for mycelial and sporophoral dispersions. The flasks were incubated at [Vor. 3 494 ANNALS OF THE MISSOURI BOTANICAL GARDEN 25-30°С. for 21 days, after which they were titrated for phos- phorie acid which was calculated as phosphorus pentoxide according to the uranium acetate method described by Hawk (12, p. 413). Опе cubic centimeter of the solution of uranium acetate used was calculated to be equivalent to 4.65 mg. of phosphorus pentoxide. Two cubic centimeters of a solution of sodium acetate were added to 10 се. of the filtered sub- strate to be titrated. "This was brought to a boil and titrated TABLE XIV NUCLEASE ACTIVITY IN LENZITES SAEPIARIA Free Н,РО, as P,O, Net Р,О, in 25 cc. Amount of enzyme dispersion after 21 days ot ecd used with 25 cc. of 1 per cent Mycelium | Sporophore | Mycelium | Sporophore mg. mg. mg mg. 5 cc. enzyme dispersion......... 40.11 59.31 28.49 37.83 5 cc. enzyme dispersion (auto- UOS RE EE 11.62 2208 7 1 X... T ss 5 cc. distilled water............. А Foe loo ee while hot. Potassium ferrocyanide was used as an indicator. Table хгу shows the results of these experiments, the figures given being the averages of duplieate experiments which were run in all cases. These results demonstrate the presence of nuclease both in the mycelial and sporophoral tissues of Lenzites saepiaria. However, no quantitative comparison can be drawn, for the mycelial dispersion was made from the mycelial mat in saw- dust. Compared with the results obtained by Kikkoji, the sporophoral dispersion is not so active as that in Cortinellus. This may be due to the fact that he used a more concentrated substrate and the juice of the fresh fungus direct. The fact that phyto-nuclein, an alpha-nucleoprotein, was used as a substrate, and that this was broken down to phos- phorie acid is a definite control on our results illustrating tryptie enzymes. The nucleoprotein must first be broken down by a nucleinase, which, according to the consensus of opinion, is a tryptie ferment. It is only after this enzyme has acted that the nucleic acid is free to be acted upon by nuclease. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 495 PROTEASE Proteolytie enzymes have been commonly demonstrated in the filamentous fungi, especially Aspergillus and Penicillium, but in the higher forms they are not so well known. The first to discover protein-digesting enzymes in the Basidiomycetes were Bourquelot and Hérissey ('95) who found that pieces of the white of an egg, which had been eooked for 10 minutes on a water bath, were changed when placed in the juice ex- pressed from sporophores of Polyporus sulphureus. The solu- tion gave a slight biuret reaction after incubating at 29-30°С. for 21 hours. Hjort (’96) found in the sap from the sporo- phores of Pleurotus ostreatus a tryptic ferment capable of digesting fibrin. It worked best in acid solutions and pro- duced leucin, tyrosin, and tryptophan. The naturally acid watery extract of the sporophores of P. sulphureus readily digested fibrin, but if neutralized or made alkaline it did not act at all. The expressed juice, weakly acidified with hydro- chloric acid or oxalic acid, digested fibrin as well as the original extract alone. After 12 hours of digestion the action was carried to peptones only. There were no amino acids present. In 1898 Bourquelot and Hérissey investigated the same action of the expressed juice of Ше sporophores of Amanita muscaria, and found it to digest nearly all of the caseinogen of skimmed milk in 4 days, after which tyrosin was present in the solution. Kohnstamm (701) investigated the proteases of Armillaria mellea, Merulius lacrymans, and Polyporus squamosus. The expressed juice from the sporophores of Armillaria mellea liquefied neutral thymol-gelatin to the ex- tent of 1 mm. in depth in 10 days. The gelatin tubes were 8 mm. in diameter. Both the mycelium and sporophores of Merulius lacrymans were used. The juice from both was equally active in liquefying gelatin, about 8 mm. in 10 days or equivalent to 0.6 ee. The active principle is thermolabile. In a 0.2 per cent solution of hydrochloric acid the extract digested fibrin to peptone but not to amino acids, while in 0.2 per cent sodium carbonate solution there was no digestion of fibrin. The juice of sporophores of P. squamosus, collected [VoL. 3 496 ANNALS OF THE MISSOURI BOTANICAL GARDEN in January and March, liquefied gelatin at the rate of 1 mm. per day for 30 days. There was the same action on fibrin as related above for Merulius lacrymans. Vines (’03) found by allowing crushed sporophoral tissue of Agaricus campestris to act on fibrin for 22 hours that there was a complete digestion to amino acids, i.e., the tissue is able to peptonize fibrin and digest the peptones. Delezenne and Mouton (708, '03*), a little later in the same year, secured widely different results. From the dried fruiting bodies of Agaricus campestris, Amanita muscaria, A. citrina, and Hypholoma fasciculare they made extracts with 0.8 per cent sodium chloride, using chloroform or toluol as antisepties. The extracts thus prepared from all of these species converted peptone to amino acids, digested gelatin and casein, but would not peptonize fibrin. These results of Delezenne and Mouton seemed so contradictory to the observations of previous workers that Vines (204) made further experiments to test their accuracy. For this work the ground pulp of the sporophores of Agaricus campestris, with the lamellae removed, was used. Providing the sporophores were mature the digestion of fibrin was evident. However, when a watery extract of the pilei was used the results were less certain, but in other experi- ments Vines found that extraets made with 2 per cent sodium chloride from fresh and dried sporophores actively digested fibrin in 1 per cent toluol or 0.2 per cent hydrocyanie acid. Sinee he found that boiled fibrin was not digested by these extraets, he suggests that the negative results of Delezenne and Mouton must be due to this error. Thus the extracts of A. campestris, prepared by Vines, contained an enzyme cap- able of peptonizing fibrin and converting the peptones and albumoses to amino acids. From this he concludes that there are two distinct proteases present, the one trypsin, the other erepsin. Buller, in 1906, confirmed the results obtained by Kohn- stamm in 1901 on the presence of proteases in the sporophores of P. squamosus. Kikkoji ('07) demonstrated the presence 1916] ZELLER—-PHYSIOLOGY OF LENZITES SAEPIARIA 497 of a protease in the sporophores of Cortinellus edodes. It acted in neutral or alkaline solutions. Rumbold (’08), in cultural studies on various wood-destroy- ing fungi, investigated the action of 13 of them on gelatin of the following constitution: 24 per cent Liebig’s beef extract; 24 per cent malt extract; 10 per cent gelatin. The reaction was adjusted in one lot by use of sodium carbonate and in an- other by the use of sodium hydroxide. She found that L. saepiaria was the only one to liquefy gelatin, and this only when sodium carbonate was used to readjust the reaction of the medium. Experiments were conducted using various substrates such as gelatin, albumin, casein, legumin, peptone, and fibrin. The enzyme preparations and fungous meal were used in some cases, while in others the growing fungus was utilized. Action on gelatin.—A 10 per cent gelatin was prepared in the same way as that used by Rumbold (708), cited above. The reaetion was adjusted with sodium earbonate and sodium hydroxide, and poured plates prepared and inoculated with mycelium of Lenzites. After a growth of 4 days a circle of gelatin 1.2 em. in diameter was liquefied in the region of the inoeulum, but this only in those plates neutralized with sodium carbonate, while there was no liquefaction in those neutralized with sodium hydroxide. In the latter the mycelium had pen- etrated the gelatin to some extent. Gelatin in the form of Mett’s tubes was digested by the enzyme dispersion of the mycelium, but sodium hydroxide did not show the same in- hibiting effect as mentioned above. Negative results were obtained with the sporophoral dispersion. These results are quite in accord with experiments of other workers who have investigated the influence of the reaction of the medium on the growth of timber-destroying fungi. In this case the results suggest that there is an effect of alkalis on the metab- olism of the fungus, i. e., that the secretion of proteolytic enzymes by the living organism is checked by the toxie effect of such active alkalis as sodium hydroxide, but when once exereted they have no inhibitory effect on the enzyme action. [Vor. 3 498 ANNALS OF THE MISSOURI BOTANICAL GARDEN Action on albumin, casein, legumin, and peptone.—The pro- teolytie action of the enzymes of the sporophoral and mycelial dispersions was tested by means of various substrates, under acid, alkaline, and neutral conditions. One per cent solutions of albumin, casein as such, as well as in the form of com- mercial *nutrose," legumin, and peptone were prepared. Casein and legumin were dissolved in N/10 NaOH. Albumin was also used in the form of Mett’s tubes. To determine the tryptic action the biuret test was em- ployed. In testing for peptone by the biuret test it is neces- sary to get rid of the native proteins, since these give a purplish blue coloration with alkaline copper sulphate, the purple blinding the fainter pink test for peptone. In order to do this ammonium sulphate is usually added in crystalline form to precipitate the higher proteins. The precipitate, however, is generally in such finely divided particles that it will not filter out with the filter papers commonly used. To overcome this difficulty the solution was filtered through bone black to remove the precipitate. The question arose whether the bone black might not absorb the peptone, and tests were made as follows: To a few cubic centimeters of 1 per cent casein solution was added a small bit of Witte peptone. Crystals of ammonium sulphate were added to precipitate the casein. After filtering through bone black the clear filtrate gave a pink color with sodium hydroxide and dilute copper sulphate. The tryptophan test was employed in testing for the action of erepsin. Wherever a test for tryptophan was given, higher proteins being used as a substrate, there was a demonstration of tryptic action as well. The tryptophan test is the produc- tion of a pink color after the addition of a few drops of glacial acetie acid and then a few drops of strong chlorine water. Experiments were set up using 10-сс. portions of Ше above- mentioned substrates in tubes. Two cubic centimeters of the enzyme dispersions were added to each tube, except the water eontrols. 'Toluol was used as an antiseptic. Each substrate was set up in series of neutral, acid, and alkaline cultures, the acidity and alkalinity being approximately N/200. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 499 Where casein, legumin, and albumin were used as substrates there was no indication of digestion in either the mycelial or sporophoral dispersion. In no case was there positive proof of peptonization. Peptone, however, was peptolyzed in neutral, alkaline, and acid solutions. If any distinction can be made there was stronger indication of tryptophan in the acid and alkaline solutions than in neutral Quantitative de- terminations were not made. Action on fibrin.—The digestion of fibrin was determined by using the method described by Reed (713). Fibrin was stained in 1 per cent Congo red, and the color fixed by im- mersion in hot water. When such fibrin is acted on by trypsin the red color is liberated into the solution. With this colored fibrin in water as a substrate, negative results were obtained with the mycelial and sporophoral dispersions. Toluol was used as an antiseptic. Another series of tubes was set up, using 6 grams of mycelial meal in 10 cc. of water and the colored fibrin as a substrate. In this case potassium cyanide was used as an antiseptic, and the results were positive. The color value in this case was difficult to judge, however, on account of the brown color imparted to the solution by the sawdust meal, but the tryptophan test confirmed the color liberation. The question arose whether potassium cyanide is a better antiseptic than toluol, use of the first-mentioned having been recommended by Vines in certain cases. Later experiments were conducted, using the pure mycelium grown on carrot juice as a source of enzymes, while potassium cyanide, toluol, and thymol-chloroform were used as antiseptics. In this series pure fibrin in water was the substrate. In all of these except two, after digestion of two weeks, the tryptophan test was obtained, the exceptions being those of the autoclaved controls and water controls. The test was slightly more dis- tinct where potassium cyanide was used as an antiseptic. The results are sufficient to demonstrate the presence of both erepsin and trypsin in the mycelium, and at least erepsin in the sporophores of L. saepiaria. [VoL. 3 500 ANNALS OF THE MISSOURI BOTANICAL GARDEN RENNET The property possessed by the juice of certain plants of causing milk to coagulate was known as early as the sixteenth century, notably in the case of Galium verum. According to Green (793), this plant is still in use at the present day for the coagulation of milk in cheese-making. Green, Oppen- heimer (710), and Euler (712) have comprehensively reviewed the literature on the occurrence of rennet in the higher plants. Oppenheimer (210, p. 317) also lists certain bacteria, such as Bacillus amylobacter, B. mesentericus var. vulgatus, and B. prodigiosus, which cause the coagulation of milk. It has been observed by a few workers that rennet is present in certain fungi, especially Aspergillus and Penicillium (Oppen- heimer, p. 317). Buller ('06) found this enzyme to be very active in the juice from the sporophores of Polyporus squa- mosus, and Bayliss ('08) reports it for Polystictus versicolor. In fact, the existence of rennet in the Basidiomycetes is con- sidered general. Gerber (709) has demonstrated its presence in 86 species. Of the wood-destroying forms, he examined Stereum purpureum Ег., Polyporus adustus Willd., P. betu- linus Bull, P. hispidus Bull, P. giganteus Pers., Trametes Bulliardi Fr., T. gibbosa Pers., T. suaveolens L., Daedalea borealis Wahlb., Hydnum repandum L., Armillaria mellea Vahl, and Lycoperdon piriforme Schaeff. Our experiments show that rennet is present in both the mycelium and sporophores of L. saepiaria. After incubating at 25-30°C. for 3 hours the coagulum was formed by the mycelial dispersion, but it took 7 hours to form a coagulum in the sporophoral dispersion. There was no coagulation in the boiled or water control after 20 hours, the toluol keeping it antiseptic. The results show that the rennet is more active in the mycelium than in the sporophores. The biological importance of this enzyme in plants is en- tirely a mystery. There arises the supposition a priori that one has to do here with some peculiar phase of protease activity, since in animal life the activity of rennin seems to be closely linked with the proteases of the gastric and pan- creatic juices. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 501 OXIDASE AND CATALASE Interest was first aroused concerning oxidases in higher fungi because of the discoloration of certain fungous tissues when exposed to the air. During the years 1895, '96, and '97 many papers appeared by Bourquelot, Bertrand, Hérissey, et al., dealing with the oxidases in the higher fungi. They found that laccase was widely distributed in the Basidiomycetes, and in the case of Boletus cyanescens, the bluing of injured spots was due to the laccase acting with the oxygen of the air on the boletol present in the tissues. Bertrand (’96) showed that the crystalline chromogen in Russula, especially Russula nigricans, was tyrosin, and in the latter the tyrosin on exposure to the air was oxidized to melanin, a black substance. Tyrosinase in the tissues oxidized the tyrosin, causing the tissues to blacken. Lutz (712), investigating the oxidases in the stipes and pilei of Gyromitra gigas and Disciotis perlata, found tyrosinase present in both species, but in both a more marked action in the eaps than in the stipes. Euler ('08) carefully investigated the catalase of Boletus scaber. There seemed to be a rela- tion between the oil content of the fungus and the amount of catalase present, and the presence of a metal, like magnesium hydroxide, in the solution increased the catalytic action. The literature concerning the function of oxidases in plants has been amply considered by Clark (711), who makes special mention of the relation of oxidases to chromogens in the higher fungi and their possible aid in the respiration process. In my experimental work on oxidases and catalase no quan- titative determinations were made. In some instances the enzyme dispersions were used, in others the fresh tissues. Clark's (710) methods were used, and guaiacum, alpha- naphthol, and paraphenylenediamine were used as indicators of oxidation. The mycelial extract was made from pure cul- tures of L. saepiaria grown on Thaxter's potato-hard agar. Ten grams of fresh fungous mat were ground with sand and treated with 100 cc. of distilled water. This extract (5 drops in 5 се. of H202) caused a rapid evolution of gas, showing the presence of catalase. The sporophoral meal showed much [VoL. 3 502 ANNALS OF THE MISSOURI BOTANICAL GARDEN greater activity than that of the mycelium. Some sporopho- ral enzyme dispersion, added to hydrogen peroxide in an evaporating dish, showed active evolution of oxygen, but the mycelial dispersion treated in the same way gave none. Oxidase action was shown by use of guaiacum and par- aphenylenediamine but not with alpha-naphthol. The ex- periments were set up in the following way, and each series in duplicate: То 5 се. of the fresh fungous extract were added 5 drops of hydrogen peroxide and 10 drops of the indi- cator. With the guaiacum a faint blue tinge was produced in 2 hours when the mycelial extract was used. When an extract from the dried sporophores was used in the same way the blue color was more distinct in 2 hours. The paraphenylene- diamine gave a brownish color in about 6 hours both in the sporophoral and mycelial extracts. Tyrosinase was demonstrated both in the sporophores and mycelium. The substrate used was a suspension of tyrosin in distilled water. To 10 ce. of this suspension 2 ce. of enzyme dispersion were added. There was no oxidation in the auto- claved controls, but in 16 hours the suspension containing mycelial dispersion had become a light gray, while that con- taining the sporophoral dispersion was a dark gray. Tyrosin when oxidized becomes black, but here where only partially oxidized it shows a gray color. The tyrosinase is more active in the fruiting bodies, as in the case of catalase and other oxidases. SUMMARY In this paper there are considered some of the more im- portant aspects of the physiology of a wood-destroying organism, Lenzites saepiaria. 1. The fungus was grown in pure cultures, and the char- acteristics of the mycelium and sporophores produced under cultural conditions are described. 2. Some factors influencing the growth and metabolism of the organism are discussed, and experimental results are given on the relations of the fungus to reaction of media, to water, and to oxygen. Special interest is attached to the in- 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 503 fluence of the resin content of the substratum on the growth of the fungus. A resin agar emulsion was prepared, and experimental data show that L. saepiaria will grow well on 50 per cent resin by weight, which is considerably more than is found in any coniferous wood. Growth is not entirely in- hibited by 85 per cent resin. 3. The metabolism of the fungus was studied through the agency of enzyme action. a. A standard method of extracting and isolating the enzymes was used, and enzyme preparations were made from vegetative and fruiting tissues. The methods com- monly used of identifying the enzymes were employed. b. Among the esterases, those acting on the esters of the lower fatty acids showed more active hydrolysis both in sporophoral and mycelial tissues than those acting on the neutral fats. с. In Ше carbohydrases, positive evidence was ob- tained of the presence of maltase, invertase, raffinase, emulsin, tannase, diastase, inulinase, ligninase, cellulase, hemicellulase, and pectinase, while negative results were obtained for the presence of pectase and lactase. d. The cyto-hydrolyzing carbohydrases were made a special study, together with their effects as demonstrated by microchemical observations on sound and decayed wood. e. The action of amidase on asparagin and acetamid was practically negligible, while urease action was very decided when the fungous tissue was used instead of the enzyme preparations. The presence of hippuricase was also demonstrated. f. The following additional enzymes have also been found: nuclease, proteinases—both tryptic and ereptic— rennetase, oxidase, and catalase. 4. A comparative study of the enzymes occurring in the sporophoral and mycelial tissues was made. As we would anticipate, this comparison shows that the important metab- olic processes are carried on in the vegetative organs. In- [VoL. 3 504 ANNALS OR THE MISSOURI BOTANICAL GARDEN deed, wherever quantitative results were obtained, or where a comparison can be accurately made, as in the case of diastase, invertase, tannase, and cellulase, the greater activity is shown in the mycelium. An exception to this, however, is the oxidases, where the greater activity is in the aporophores, ACKNOWLEDGMENTS The writer acknowledges his indebtedness to the Southern Pine Association, who through their appropriation of funds made this investigation possible, to the Missouri Botanical Garden for library and laboratory facilities, and to Professor B. M. Duggar and Dr. Hermann von Schrenk for their advice and helpful criticisms. BIBLIOGRAPHY Appel, O. (15). The relations between scientific botany and phytopathology. Ann, Mo, Bot. Gard. 2; 275-285. 1915. de Bary, A. (86). Ueber einige Sclerotinien und Sclerotien-krankheiten. Bot, Zeit. 44: ST дет, 393-404, 409-426, 433-441, 449-461, 465-474. 1886. Bayliss, Jessie S. (08). The MES of Polystictus versicolor (Fries). Jour. Econ. Biol. 3: 1-24. pl. 1-2. 1908. Bernthsen, A. (12). А textbook of ` ¿rm chemistry. pp. 1-720. 1912, [Edited and revised by J. J. Sudborough.] Bertrand, С. (96). Sur une nouvelle oxydase, ou ferment m —— d'origine végétale. Compt. rend. Acad. Paris 122: 1215-1217. (962). Sur la présence simultanée de la laccase et de la tyrosinase dans la suc de quelques champignons. Ibid. 123: 463-465. 1896. Biffen, R. H. (99). А fat-destroying fungus. Ann. Bot. 13: 363-376. pl. 1. 1899. ------ (01). On the biology of Bulgaria polymorpha Wett. Ibid. 15:119- 134. pl. 7. 1901. Bloor, W. R. (14). A method for the determination of fat in small amounts in blood. Jour. Biol. Chem. 17:377—384. 1914. Bourquelot, E. (93). Les ferments solubles de l'Aspergillus niger. Soc. Мус. Fr. Bul. 9: 230-238. 1893 résence d'u un ferment analogue à l'emulsine dans les cham- (9 pignons et en particulier dans ceux qui sont parasites des arbres ou vivent sur le bois. Ibid. 10:49—54. 1894. ———, (96). Des composes oxydables sous l'influence du ferment oxydant des champignons, Compt. rend. Acad. Paris 123: 315—317. 1896. ------, (Оба). Action du ferment soluble oxydant des champignons sur les phénols insolubles dans l'eau. Ibid. 123:423—425. 1896. ------, (97). Sur la présence ee m les champignons, d'un ferment oxydant gring: sur la tyros ur mécanisme de la coloration du chapeau de ces végétaux, Soc, Муо. Tr, "Bul 13: 65-72. 1897. 1916] ZELLER—TPHYSIOLOGY OF LENZITES SAEPIARIA 505 ------- et Bertrand, G. (796). Les ferments oxydants dans les champignons. Ibid. 12; 18-26. 1896. —— ——, —s (96a). Sur la coloration des tissus et du sue de certains champignons au contact de l'air. Ibid. 12: 27-32, 1896. — et Harley, V. (96). Sur la recherche et la présence de la tyrosine dans quelques champignons. Ibid. 12: 153-156. 1890. — et Hérissey, H. (95). Тез ferments solubles du Polyporus sulfureus (Bull.). Ibid. 11: 235-239, 1895. -------) ————, (98). Recherche et présence d'un eg soluble protéo- hydrolytique dans les champignons. Compt. rend. Soc. Biol. X. 5; 972-974. , et Marchadier, L. (04). Étude de la réaction provoquée par un ferment oxydant indirect (anaéroxydase). Compt. rend. Acad. Paris 138: 1432-1434. 1904. Brown, W. (15). Studies m the physiology of parasitism. I. The action of Botrytis cinerea, Ann. Bot. 29:313-348. 1915. Buller, A. H. R. (05). The destruction of сомен paving blocks by the fungus, Lentinus lepideus Fr. Jour. Econ. Biol. 1:1-12. 1905. —— ———, (06). The enzymes of Polyporus squamosus Huds. Ann. Bot. 20: 49-59. 1906. —— ———, (7062). The biolo ogy of Polyporus squamosus Huds., a timber-destroying fungus, 52 Econ. Biol. 1; 101-138. pl. 5-9. 1906. ; (09). Researches on fungi. pp. 1-287. pl. 1-5. f. 1-83. 1909. Butkewitsch, W. ('03). Umwandlung der Eiweissstoffe durch die niederen Pilze im Zusammenhan ange mit einigen Bedingungen ihrer Entwickelung. Jahrb. f. wiss, Bot. 38: 147-240. 1903. Clark, E. D. (10). The plant oxidases. pp. 1-111. 1910. -------, (11). The nature and the "Tv of the plant oxidases, Torreya 11: 23-31, 55-61, 84-92, 101-110. 191 Cooley, J. S. (14). A study of the E n Sa RW "Egeter cinerea, (Bon.) Schröter. Ann. Mo. Bot. Gard. 1: 291-326. Cousin, H., et Hérissey, H. ('08). Oxydation de l'eugénol par le ferment oxydant es champignons et par le perchlorure de fer; obtention de dehydrodieugénol. Compt. rend. Acad. Paris 146: 1413-1415. 1908. Cross, C. F., and Bevan, E. J, (01). Researches on cellulose, I, 1895-1900. pp. 1-180. 1901. ,—— (206). Ibid. II, 1900-1905. pp. 1-184. 1906. ————, — (12). Ibid. ІП, 1905-1910. pp. 1-173. 1912. Czapek, Е. (99). Ueber die sogenannten Ligninreaction des Holzes. Zeitschr. f. physiol, Chem. 27; 141-166. 1899. —————, (99a). Zur Biologie der holzbewohnenden Pilze. Ber. d. deut. bot. Ges. V: 166-170. 1899. ------- (13). Biochemie der Pflanzen. 1:1-828. Jena, 1913. [See рр. 682- 694.] Davis, A. R. (15). Enzyme action in the marine algae. Ann. Mo. Bot. Gard. 2:771-886. 1915. [VoL. 3 506 ANNALS OF THE MISSOURI BOTANICAL GARDEN Dean, A. L. ('03). Experimental studies on inulase. Bot. Gaz. 35: 24-35. 1903. Delezenne, C., et Mouton, H. (08). Sur la présence d'une kinase dans quelques ampignons Basidiomycetes, Compt. rend. Acad. Paris 136: 167-169. 1903. -----, (7034). Sur la présence d'une erepsine dans les champignons Basidio- mycetes. Ibid. 136: 633-635. 1903. Dox, A. W. (09). The intracellular EC of lower fungi, especially those of Penicillium Camemberti. Jour. Biol. Chem. 6:461-467. 1909. — — ——, (710). The intracellular enzymes of Penicillium and Aspergillus. U. 8. Dept. Agr., Bur. An. Ind. Bul. 120: 1-70. 1910. — ——, und Neidig, E. (13). Enzymatische Spaltung Wer LR durch белшде. Zeitsehr. f. physiol. Chem. 85:68—71. 1913. Duggar, x E (05). The principles of mushroom growing and кас spawn U. S. Dept. Agr., Bur. Pl. Ind. Bul. 85: 1-60. pl. 1-7. Euler, H. (00), Т” Kenntnis der Katalase, Beitr. z. chem. Physiol. u. Path. 7:1-15. 190 ------, (19). General eR of the enzymes. pp. 1-323. 1912. [Transla- tion from German by Thos. H. Pope.] Falek, R. (02). Die Cultur der Oidien und ihre Ruckfuhrung in die hohere Fruchtform bei den Basidiomyceten. Beitr. z. Biol. d. Pflanzen 8; 307-346. pl. 12-17. 02. — (709 Die Lenzites- — ә SCT Moller's Haussehwamm- forschungen. Heft 3: 1-234. p 7. f. 24. Jena, 1909. , (719). Die Merulius-füule des Bauholzes. Ibid. Heft 6; 1-405. pl. 1-17. f. 73. Jena, 1912. Ferguson, Margaret C. (702). A preliminary study of the EC of the spores of Agaricus campestris and other basidiomycetous fungi. U. 8. Dept. Agr., Bur. Pl. Ind. Bul. 16: 1-43. pl. 1-3. Gerber, M. C. (709). La présure des Basidiomycetes. Compt. rend. Acad. Paris 149: 944-947, 1909. Grafe, V. (704). Miss doe erf über die Holzsubstanz vom m ios ngs- logischen Standpunkte Wiss., Wien, math.-naturw. Kl ber. 113:253-295. 1904 Green, J. В. ( On the germination of the tuber “ aa Jerusalem artichoke я 788). (Helianthus tuberosus). Ann. Bot. 1:223-236. — ————, (93). On vegetable ferments. Ibid. 7:83-137. 1893. Hartig, R. (78). Die Zersetzungscheinungen Di Holzes der Nadelholzbiiume und der Eiche. pp. 1-151, pl. 1-21. Berlin, 187 Hawk, P. B. (12). Practical physiological deis. pp. 1-475. 1912. — Dr. (11). Notiz ueber den Cellulosegehalt von Eichenholz, ханы са helephora Е veründert war. Naturwiss. Zeitschr. f. Forst- u. Landw Š: 246-250. 191 Hjort, 7. (96). Neue eiweissverdauende Enzyme. Centralbl. f. Physiol. 10: 192-194. 1896. Humphrey, C. J., and Fleming, Ruth M. (15). The toxicity to fungi of various oils and ай», particularly those used in wood preservation. U. 8, Dept. Agr., Bul. 227: 1-38. pl. 1-4. 1915. 1916] ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 507 Iwanoff, L. ('03). Ueber die fermentative Zersetzung der a durch Schimmelpilze. Zeitschr. f. physiol. Chem. 39; 31-43. 1903. Jean, Е. (00). Die Bestimmung des Tannins und der Gallussüure. Ann. Chim anal, appl. 5: 134-140. 1900. [Chem. Centralbl. 1900': 1107-1108. 19001 m J. B. (93). Timber physics. Investigations on long-leaf pine. 4. Re- on mechanical tests. U. S. Dept. Agr., For. Div. Bul. 8: 22-31. f. 11-16. Jones, L. R. ('05). The eytolytie enzymes produced by Bacillus carotovorus and certain other soft rot bacteria. Centralbl. f. Bakt. II. 14: 257-272. 1905. Kellerman, K. F., and McBeth, I. С. (12). The fermentation of cellulose. Ibid. 34: 485—494. pl. 1-2. 1912 Kikkoji, T. C07). Ueber das Vorkommen von einem Nucleinsiiurespaltenden Fermente in Cortinellus edodes. Zeitschr. f. physiol. Chem. 51:201-206. 1907. Knudson, L. (13). Tannie acid fermentation. I. Jour. Biol. Chem. 14; 159-184. f. 1-2. 1913. --------, (13a). Ibid. II. = y nutrition on the production of the enzyme, tannase. Ibid. 14:185-202. Kohnstamm, P. (701). Amylolytische, — proteolytische und Cellu- lose lósende Fermente in holzbewohnenden Pilzen. Bot. Centralbl., Beih. 10: 90-121. 1901. Kossowiez, A. (12). Die enzymatische Natur der Натпвйлге- und Hippursüure- Garung. Zeitschr. f. Garungsphysiol. 1; 121-123, 317-319. 1912. Le — А. (719). Influence du milieu sur la résistance du Pénicille crustacé ux substances toxiques. Ann. Sei. Nat., Bot. IX. 16: 277-336, 1912. Lutz, M. L. (12). Sur la гбвепсе dans le Gyromitra gigas et le Disciotis per- lata de tyrosinase et d'un chromogene. Soc. Myc. Fr. Bul. 28: 136-199. 1912. Lyman, G. В. (07). Culture studies on poly He of Hymenomycetes. Boston бос. Nat. Hist., Proc. 33: 125-209. pl. 18-26. 1907. McBeth, I. G., and Scales, F. M. кы The destruction of cellulose by bacteria and filamentous fungi. U. S. Dept. Agr., Bur. Pl. Ind. Bul. 266; 1-52. pl. 1- Mangin, L. (92). Propriétés et um des composés peetiques. Jour. de Bot. 6:206-212, 235-244, 363-368. -----, (93). Ibid. SC 121-131, 325-343. pl. 1-2. 1893. Miyoshi, M. (95). Die Durc EE von Membranen durch Pilzefiiden. Jahrb. f. wiss. Bot. 28: 269-289. e? 1-3. 1895 Münch, E. (709). Untersuchungen über Immunität und Krankheitsempfüng- liehtkeit der Holzpflanzen. Naturwiss. Zeitschr. f. Forst- u. Landw. 7: 54-75, 87-114, 129-160. f. 1-5. 1909. Newcombe, Е. C. (99). Cellulose-enzymes. Ann. Bot. 13: 49-81. 1899. Oppenheimer, C. (^10). Die Fermente und ihre Wirkungen. pp. 1-606. 1910. Pringsheim, Н. (08). Ueber Pilzdesamidase. Biochem. Zeitschr. 12: 15-25. 1908. Reed, H. S. (13). The enzyme activities involved in certain fruit diseases. Va. Polytec. Inst., Agr. Exp. Sta. Rept. 1911-1912:51-77. 1913. [VoL. 3 508 ANNALS OF THE MISSOURI BOTANICAL GARDEN Richards, H. M. (’97). Die NE des Wachsthums einiger Pilze durch chemische Reize. Jahrb. f. wiss. Bot. 30: 665-688. 1897. Rumbold, C. (708). Beiträge zur Kenntnis der Biologie holzzerstórender Pilze. Na turwiss, Zeitschr. f. Forst- u. Landw. 6: 81-140. pl. 1. f. 1-22. 1908. Schellenberg, H. С. (708). Kee meet das Verhalten einige Pilze gegen Hemicellulosen. Flora 98: 257-308. von Schrenk, Н. ('00). Two diseases a red cedar caused by Polyporus juniperinus n. 8p. and Polyporus carneus Nees. U. Б, Dept. Agr., Veg. Physiol. and Path. Bul. 21: 1-22. pl. 1-7. f. 1-3. 900. -----» (7004). Some diseases of New England conifers. Ibid. 25: 1-56. pl. 1-15. f. 1-3. 1900. (700 A disease of Taxodium distichum known as peckiness, also a similar disease of Libocedrus decurrens known as pin-rot. Mo. Bot. Gard Rept. 11:23-77. pl. 1-6. 1900. —— (701). A disease of the black locust (Robinia Pseudacacia L.). Ibid. 12: 21- 31. pl. 1-3. 1901 » (03). A disease of the white ash caused Kë PI fraxinophilus. U. S. Dept. Agr., Bur. Pl. Ind. Bul. 32: 1-20. pl. 1 190 — (14). Two OM diseases of the mesquite. Ann. Mo. Bot. Gard. 1: 243-252. pl. 6-7. ------- (143). A trunk disease of the lilac. Ibid. 253-202. pl. 8-9. 1914. Schwalbe, C. G. (11). Die Chemie der Cellulos а даа, besonderer Beriicksichti- ung der Textil- und Zellstoffindustrien. Berlin, 1911. Shaffer, P. A. (14). On the determination of sugar in blood, Jour. Biol. Chem. 19:285-295. 1914. Shibata, K. (04). Ueber das Vorkommen von Amide ола Enzymen bei Pilzen. Beitr. г. chem. Physiol, u. Path. 5; 384-394, 1904. Singer, M. (82). Beiträge zur naheren Kenntnis der Holzsubstanz und der verholzten Gewebe. K. Akad. Wiss., Wien, math.-naturw. K1., — 86: 345- 1882. (Cited by Grafe, '04, p. 357.) GR Sg E (11). The timber rot caused by Lenzites saepiaria. U. S. Dept. Agr., . Pl. Ind. Bul. 214: 1-46. pl. 1-4. f. 1-3. 1911. Vines, S. Н. (03). Proteolytic enzymes in plants. II. Ann. Bot. 17: 297-316, 1903. Ward, H. M. (88). А lily-disease. Ibid. 2:319—382. pl, 20-2). 1888. — — ——, (97). On the biology of Stereum hirsutum Fr. Roy. Soc. London, Phil. Trans. 189°: 123-134. pl. 17-21. 1897. — Дай Penicillium as a wood-destroying fungus. Ann. Bot. 12:505- 566. Weer C. (12). Hausschwammstudien. I. Zur Biologie von "eye та cere- a А. & Sch. Мус, Centralbl. 1: 2-10; Ibid. ПТ. 2: 331-340. 1912 —— (14). Ibid. IV. Versuche ueber die Ары err der Holzansteckung und -Zersetzung durch Merulius. 1014. 3; 321-332. f. 1; Ibid. V. 4: 241-252. f. 1; 281-299. pl. 1-2. 1914. — — —, (Ma). Holzansteckungsversuche mit we Trametes, und Polyp- orus, We d. deut. bot. Ges. 32: 566-570, 1914 ZELLER—PHYSIOLOGY OF LENZITES SAEPIARIA 509 ------ (14). Die chemische Wirkung des Hausschwamm auf die Holzsub- stanz. Ibid. 32: 601—608. 1914. Weir, J. R. ('14). Notes on wood-destroying fungi which grow on both coniferous and deciduous trees. І. Phytopath. 4; 271-276, 1914. Westerdijk, Johanna (15). Phytopathology in the tropics. Ann. Mo. Bot. Gard. 2: 307-313. 1915. Wolf, F. A. (16). Citrus canker. Jour. Agr. Res. 6: 69-99. pl. 8-11. f. 1-8. Wolff, J. (209). Sur quelques propriétés = des oxydases de Russula Delica. Compt. rend. Acad. Paris 148; 500—502. 510 [Vor. 3, 1916] ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 8 Figs. 1, 3, 4, 6, and 7. Sporophores of Lenzites saepiaria grow- ing on sawdust in pure culture, and showing the thelephoroid, daed- aloid, and irpiciform characters produced under cultural conditions. Fig. 2. Sporophores in situ on blocks of Pinus echinata in pure culture. Fig. 5. The lower surface of a sporophore borne on a horizontal surface of a cultural block. The hymenium is borne on typica Lenzites gills. igs. 8-12. Samples of pine after one year in culture under favorable moisture conditions, showing the typical internal decay. Figs. 13-17. Samples of pine showing the superficial “scorching” after one year in culture, during which time the blocks were sat- urated with water. PLATE 8 14 -LENZITES SAEPIARIA SDLÉER 4 Zl [Vor. 3, 1916] 512 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 9 To illustrate the growth of Lenzites saepiaria on resin agar plates in Petri dishes of uniform size in 14 days. Fig. 1. Control plate showing the growth on a Thaxter’s glucose- potato-hard agar containing no resin. Figs. 2-21. Showing the growth on 5-100 per cent resin agar, Ең goad (each plate increased by 5 per cent resin AUR T e Thaxter's glucose-potato-hard agar was used as ANN. Mo. Bor. GARD., VOL. 3, 1916 PLATE 9 ZELLER—LENZITES SAEPIARIA GENERAL INDEX TO VOLUME III New scientifie names of plants e£: the ph i of new combinations e? d mbers having reference to figure yms and page or plates, in italic; and previously о ибей e names and all other ien? type. a i in bold face type; synon in ordinary A Дренов Schénleinii, eg Acrostalagmus albus Agrimonia, 309; Ee 313; Eupa- ‚ 313, 315, 316, 317, var. ameri- me 3 r. glabra. , 313, var. suta, 313, ar. mollis, 315, var. parvi- flora, 313; gryposepala, 313; hirsuta, 13; microcarpa, 315; 48, 316; parviflora, 317; parviflora, 313, 316 KEE 315; polyphylla, 317; pubescens, 315; pum ros- 1 A tellata, 313; semrifotia, ee striata, 13; suaveolen Agrimonies, The , 309 Alternaria sp., ian ob 279, 281 Amidases in Lenzites saepiaria, 489; in other fungi, Apparatus, à no eie the adaptability of the Folin ae Kjeldahl, for tk, 407 (Thelephora), d? argillaceus (Hypochnus) , icc S re- Aspergillus, 2 271; aviarius, 253, 271; д Байы 254, 271; candidus, 254, 271; flavescen 8, 254; flavus, ee: fontoynonti, 255, 271; fum obaeter chroococcum, nitrogen fixa- i 3, 431; vinelandii, nitro- gen fixation in, 423, B Basidiomycetes, temperature relations of the, Ai bicolor (Hypochnus), 229 ANN. Мо. Вот. GARD., VoL. 3, 1916 bicolor Ad biennis (Thelephora), 213 botryoides met" Gë botryoides iur ес tryosporium, 22; pyr i eren Botryosp Mondiale, 22 Botrytis cinerea Ke E. Disch (subg. Pistil- a) Burt n. sp., 403; The Tilephoraseae of North der Bus 5 SS The Missouri agrimonies, 309 С Cabbage yellows, 25; and the relation of temperature to its occurrence, 25; causal organism of, 28; symptoms of, 27 canadensis ( Hypochnu в), 211 33 in rg а es ER 501; in ther fugi, $ Catalogu e of the pM of Jasper ounty, Missouri, Cellulass i in Lenzites poro 474; in other fungi, 471 Baesch, (Hypochnus), 232 alate ie (Zygodesmus), 219 Cineraria lyrata, 99 унав, " yp 233 cirratum (Septobasidium ) , 994 Claudopus subnidulans, 195 Conioph ra, 204 $ БЕ coriarius M cree orticium, sehinosporun, 237; EUR итови 239; sul- 208, f fuscum 215; isabellinum, 292; lk ege 239; thelephoroides, 235; недрата 213 Cotton root rot, disease of, 11 Cryptococeus 203, 2125 Corsellii, 263; degeneran ; farciminosus, 4; Gilehristi, "ota: of Gotti & Brazzola, atogenes ; hom саке иа, 264; litho- 5; niger, 265; Tokishi- gei gh - 262 Cyto- hydrayzing enzymes, 465; effect of, und and de ecayed wood, 484 (513) 514 D Davis, A. R.: A note on the еее Cé ity o ; Dug- , Studies in the physi- gar, ә ап ology of the fungi. I, Nitrogen Fixa- tion, 413 Diastase in Lenzites saepiaria, 464; in other fungi, 464 канот р Solani іп апа the A as root and its соса! stage, 11; апа Devis A. R., Studies in the phys- iology of the fungi. I, Nitrogen Fixa- tion, 413 E echinosporum (Corticium), 237 —— соттотом), 237 elaeodes (Hypochnus), mig, W. H. eimi in natur = certain re -like fungi with las ce to thei d tegt Ia in e higher animals, Emulsin in Lenzites Gen 462; in other fun albicans, 259, 272; tropicalis, Enzyme activity in Lenzites saepiaria, 452 = En (Hypochnus), 2 226 Esterases in Lenzites saepiaria, 455; in other fungi, 455 F р (Thelephora), 208 ferru nea (Tomentella), 208 Folin and Farmer, а of E dahl method of nitrogen determ tion by, 408; apparatus used in, 407, Fórster method of determining nitro- gen frustulosa (Hymenochaete) frustulosum ypochnus), Gem atum "Septobusidium) wä fumosum (Corticium), свечан (Hypochnus), es [Vor. 3 ANNALS OF THE MISSOURI BOTANICAL GARDEN Fungi: gill, New or interesting species of, from Missouri, 195; Studies in certain, with reference to thei possible pathogenicity i in the haber animals Fungi е 7 263, e tempera- ture relations of the Fusarium, 25; жа. 28, 1 ep shame ne? of, 56, host relations of, 43, ulation experiments with, 35, e relations viru- lence of cultures of, 39; orthoceras, 30 изса (Odontia), 239 fusca (Thelephora), 215 — (dr — i. 215 fuscata (Hypochnopsis) , 213 us), 213 fuscum (Co m), 215 mer Шол. 215 G Gill fungi, Е = interesting species of, from Miss 195 Gilman, J. C. MUN тант ала € — ы temperature to its oe fuscatus (Нур ме Бет геп дон 270, 272; Graphii, 270, er Glomerella Gossypii, nitrogen fixati in, 424, 431 Grandinia, 204; coriaria, 228; tabacina, 218 granosa (Thelephora), 227 sus (Hypochnus), 227 granulosus (Hypochnus), 218 granulosus (Zygodesmus), 21 Greenman, J. M. new Senecio f Jamaica, 201; Monograph of the North E Central American Species he us етекте П, 85 e web: 67 H ee dien A in Lenzites saepiaria, 479 Hippurica e in ES saepiaria, 492; in “qanraq права (Pistillina), 4 Hymenochaete e Zoe 337 Hymenomycete, the smallest known, 403 Hypochnus, 203; argillaceus, 222; aurantiacus, ae avellaneus, 225; bicolor, 229; tryoides, 226; canadensis, 211; eg erascens, ; cori- arius, 228 echinosporus, 237; elaeodes, 218; epigaeus, 226; ferru- gine ; ferruginosus, 212; brillosus, 238; fuligineus, 232; fulvo-cinctus, 228; fumosus, 239; 1916] INDEX fuscatus, 213; пара de granosus, 227; granulos 218; sabellinus, e ie? 8, d ensis, 13; thelephoroides, 235; tristis, ; umbrinus, 213; zygodesmoides, 236 Hypochnopsis fuscata, 213 I Inulinase in wt saepiaria, 488; in other plants, 488 zites GEN 459 isabellinum (Corticium т), 2 isabellinus (Hypochnus), 222 J jamaicaense (Septobasidium), 333 poar ounty, Missouri, description of, 349; plants of, 359 Jola, 319 K pum method of determining nitro- the Folin modification of the Seifert, 407 L Lactase in Lenzites saepiaria, 458 reg (Septobasidium), 335 s saepiaria Fries, 439; growth of, on resin k plates, 612; pine blocks decayed by, 510; ’ sporophores of, 510; with special reference to enzyme Дена на 439 Тертеиги (Septobasidium), 332 ton, leprosus (Hypochnus), 223 Ligninase in Lenzites saepiaria, 469; ngi, lilacinum (Septobasidium), 343 M gyros -Kjeldahl method of determining itrogen, 410, 426 Конаш commune, nitrogen fixa- tion in, 423, 430 Maltase in Lenzites saepiaria, 457 Moniliopsis Aderholdii, “Mopopilz” and the “vermehrungspilz,” Rhizoctonia Solani in relation to the, 1 515 Mortierella, 253, 271 Mucor, 250, 270; cornealis, 250, 270; 70; Mucedo, 251, готи " mosus, 252, ; Regnier ri, 951,” 271; mesi 252, EN Truchisi, 251, 271 N pee fixation, 413; methods of de- ing amount of, 426; organisms el in exper осе іп, 493; results of experiments in, 429 Nuclease in Lenzites saepiaria, 493; in other fungi, 493 O obscuratus (Hypochnus), 216 Odontia, 204; fusca, 239; porriginis, vacea B botryoides (Thelephora), 226 220 “qaqas 7 (Нурос 5 olivascens (Zygodesmus), 221 Oos , 266, 272; buccalis, 267; lin- , 20 porri A Б 1 monalis, 266; sp., cultur 281; а. 268; of 2.84 267 tomyces purpureus, 256 Orie Its, L. O. New or interesting species ‘of gill fungi from Missouri, 9 Oxidases in Lenzites saepiaria, 501; in other fungi, 501 Ozonium, 11; auricomum, 11; omni- vorum, 11 P Palmer, E. J. E и. the plants , Missouri, 345 o Panaeolus Аут. atus, 195, 200; rufus, 196, 200; variabilis, 197, 200 Patouillardii ( WV EE F Pectase га — "m saepiaria, 483; in other fungi, he mea Geh Lenzites saepiaria, 483; in other Steg Яв рога), 323, pos 327 pedicellatum ( — ) А Penicillium digitatum, nitrogen фа іп, 423, 431; сен nitrogen fixa- tion in, be Peniophor peniophorcides (Hypochnus) 234 Phlebia vaga, homa Betae, diio fixation in, 423, 31 Өл... [Vor. 3 516 ANNALS OF THE MISSOURI BOTANICAL GARDEN Phycomycetes, ae E temperature relations of the je жемес EE Ga ; omnivorum, - t plan < = 12, mycelium of, amida pilosus ze uites 22] Pinus авт ata, 453; palustris, 453; Taeda, 453 Pistillaria азығы Е том hyalina, plu RENT So oe ell (Sebacina), 241 Pluteus cervinus var. caespitosus, 199, Protease in Lenzites saepiaria, 497; in other fungi, 495 pseudopedicellatum (Septobasidium ) , punicea (Thelephora), 229 R Raffinase in Lenzites saepiaria, 460; in other fungi, 46 Rennet i n Lenzites saepiaria, 500; in cud geg Soria (Be (Sepiobasidium), ies срне ( така га), Rhinotri , 22 Rhizoctonia "башы іп relation to the mopo pilz” and the “vermehrungs- pilz, Rhizomucor parasiticus, 251; septatus, Rhizopus, 253, 271; Cohnii, 253, 271; niger, 253, 271 ; nigricans, 252, ?11 i 909 rubiginosus (Hypochnus), 209 rubiginosus (Zygodesmus), 209 S — Өр: 257; anginae, 257, 272; $T culture of, 284; granu- Бен Pee Па АА 258, 272; guttulatus, 258, Sehizomycetes, temperature relations o Sc hweinitzi (Septobasidium), 324 Sclerotin Beie geg 241; plumbescens, Senecio, A new, from Jamaica, 201 Monograph of the North and Conta American Species of the genus—Part II, 8 Senecio, 85; acutidens, 178; Adamsi, 172; anacletus, 183, i i leticu = ` m a = m 4 ; Ф 119, var. gracilis, 128, var. semi- cordatus, 129; aureus, 91, 129, var. ате 155, var Balsa itae, 130, bed 155, 159, Aer var. ене 179, "уал. croceus, 142, 144, 173, var. discoidea, 113. compactus, 179; 179; conterminus, zit Orav fordii 139; crocatus, 141 Wolfii 3; cro- catus, 145; Шуй ана и; суш- орат, 174, var. borealis, 177, var. streptanthifolius, 177; cym- ромеи 104, 173, 178, var. diver- 171; debilis , 96; densus, 179; robek ки. 144 ; di scoideus, 90, 91, 97; Earles, 155; Elliottu, 108; elongatus, , 113; Farriae, В: fastigiatus, 119; fedifolius, 90; Fernaldii, 90; flavovir rens, 167, var thomsoniensis, 169; ovirena, 96; rt anus, 147; hesperis, 102; hesperius, 102, 190; dee 144; Hollickii, 201; hyperborealis, а hyperborealis var. columbie ; idahoensis, 96; Saget ai 91; ones E at laeti- flor 170; erti, 91; Lind heim 114; Lies 8, I SI 101; manitove ensis, 179; Me mingeri, 155; Metcalfei, "on, micro- dontus, 7183; ал ак: 169; tabilis, Mt а из, eri e obla combei nceolatus, 19; obovatus, 164 var. divisifolius, 112, : f ; pauperculus var. firmifoliu i», 166; ЕШ "M8; рікті 141; Port 106; prionophyllus, 140; “¿aska asqa 132, var. ese M pseudotomentosus, 149; pyroloi СА m - nella, 107; swavis, 179; subcuneatus, 174; subnudus, 103; Suksdorfii, 172; 1916] INDEX 517 SM E 201; toluecanus, 186, 194, ar. modestus, 188; Toluccanus var. crodontus, 183; Tracyi, 142; tri- dentieulatus, 179; tussilaginoides, 119; Wardii, 183, 190; Willingii, 54; Wootonii, 183 Septobasidium, 319; atratum, 334; can- escens, 342; castaneum, 330; cir- ratum, 334; frustulosum, 337; fumi- 324; Spongia, 339; sublilacinum, 26 ом (Hypochnus), 213 s (Hypochnus), 225 spiniferus (Hypochnus), 218 a (Septobasidium de SE oe ( Thelephora), 3 o. (Thelephora), ze spongios Маси us), 2 Spo tale 268, 272; 5 Audouini, 266, 268; Furfur, 269; minutissim Steri Cen 257, “q nigra, 257, 272; pseudonigra, Studies in the den of the fungi. I, Nitrogen Fixation, 413; n- zites diei with special reference , 931 ublilacinum (Septobasidium), 331 inosus (Hypochnus), 231 sulphureum (Corticium), 239 T tabacinus (Hypochnus), 218 Tannase in Lenzites saepiaria, 463; in other fungi, 463 Temperature, те: relation of: to the urrence of cabbage yellows, 32; to the Ascomycetes, 51; to the Ba- sidiomycetes, 53; to the Fungi Im- perfecti, 52; to the Phycomycetes, 49; to the Se chizomycetes, 49 Texas root +: ғара and its conidial staee, The, Thaxteri шина), 406 Thelephor ; arachnoidea, 213; biennis, "513; botryoides, 226; ferru- , 208; floridana, 214; fusca, 215; granosa, 227; olivacea botryoides, 226; pedicellata, 323, 324, 327; punicea, 229; retiformis, 338; Spongia, 339; — 216; su ub- Шаота, 331; wm 213; vinosa, 215; zygode. биде, 236 Тер аат. of North America, The, VII, 319 14 Tomentella, 203; aurantiaca, 941; cin- erascens, ; ferruginea, 208; Жам 215; rubiginosa, 209; tristis ‚ 213 T'omentellina a, 203; erruginosa, 212 Torula neoformans, 261; sp., cultures of, 275, tristis Ke 2 tristis (Tomentella), tropicale (Septobasidium), 326 U umbrina (Thelephora), 213 umbrinum (Corticium), 213 umbrinus (Hypochnus Urease in a other fungi, 4 s), 213 saepiaria, 489; in V vaga (Phlebia), 239 “Vermehrungspilz” and the “mopopilz,” the relation of Бойон Solani to the, Verticillium Graphii, 270 vinosa (Thelephora), 215 Y жемі = fungi, The pas in e of certain, with refer to their e d | in | higher animals, 243 Z Zeller, S. M. Studies in чи аборта: of the fungi. II, Lenzites saepia Fries, with special S teu © i се еп Zygodesmus, 204 oruber, 280; bi- color, 227, big eg 219; granu- — , 218; olivascens, 221; pannosus, 223; rubiginosus, 11 оди ei (Hypochnus), 236 eygodesmoides (Thelephora), 236 . ,. ба aditu e ED E а SC et Be = Contents | "ënnen. 3 wa Fungus and I С: Conidial gen oi в. M. Pepe “Jt: ürrence:..... Ee 212-0 пан “Monograph PE de "m th nd: Сана American Es . Species of the Genus Senecio—Part П.. 27. м. Greenman тет Interesting : Puce. ‘of LEE. zent ной. {550 ара АРМАНЫ Вт. NOTIS, EE, ES E zt E + ` Entered d matter À ák the Pos t Omce at Columbia, Missouri, under the of, M arch 3, 1879. e EN NY 4 ‘ е x & Ké > vex cei А a E Ар, £ Lat ww ke, 7% x z `x ж е калады ут; + е 3 2 d SCH 9.5 e Y a +: T Ж К = - l à “54 Cas 4 = г. M X ?? < d s 7 к Ay REC 2 ` ` " у: ` "A vi ` " xj 3 St Ke ү ж; E ud > aes UU; А rz т, v к | sour Botanical . 71-:10- 11- 23 =. м + em SE oh, o. Overholts. 195-200. PUBLISHED QUARTERLY BY THE BOARD OF TRUSTEES op THÉ маввоовт x с; E x ЖА OM қ ааа iE T AX tT 1% Ka 2; жара Ғғ У D WE Met дул `a De i ^Y dan ае 3X Tory. 5; УЭ i Мрака