(2 ANNALS OF THE MISSOURI BOTANICAL GARDEN — " РА већ) » ч и зї жээ — P еді бе ЭР © Bots » 1 tae 5 Aat me. ме. оди: бж "9 ~ ». Wb ` ч Je ag жж», + ХАТ * Л Ка md pA RR ERA, pe " =i E E 4 4 » У > РУМИ РЕ d NEU Annals of the Missouri Botanical Garden Volume XV 1928 With Fifty-three Plates and Thirty-seven Figures Published pey. at 8 West King Street, Lancaster, Pa., by the Board of Trustees he Missouri Botanical Garden, St. Louis, Mo. Entered as Ея — at the Post-Office " ae Pennsylvania, r the act of March 3, Annals of the Missouri Botanical Garden A Quarterly Journal containing Scientific Contributions from the Missouri Botanical Garden and the Graduate Labora- tory of the Henry Shaw School of Botany of Washington Uni- versity in affiliation with the Missouri Botanical Garden. Information The Annals of the Missouri Botanical Garden appears four times during 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 accept foreign subscriptions: Wheldon & Wesley, 2, 3 and 4 Arthur St., New Oxford St., London, W. C. 2, England. —— (T STAFF OF THE MISSOURI BOTANICAL GARDEN Director, Сбковавк T. Moonx. Assistant to the Director, KATHERINE Н. Lerau. HERMANN VON SCHRENK, Ernest 8. REYNOLDS, Pathologist. Physiologist. Jesse М. GREENMAN, Davip H. LINDER, Curator of the Herbarium. Mycologist. Когакр У. L. La GARDE, EDGAR ANDERSON, Geneticist. Research Assistant NELL С. HORNER Librarian and Editor of Publications. BOARD OF TRUSTEES OF THE MISSOURI BOTANICAL GARDEN President, Grorae C. Нитснсоск. Vice-President, SAMUEL C. Davis. Second Vice-President, DANIEL К. CATLIN. L. RAY CARTER. Рнпль C. SCANLAN. Tuomas S. MAFFITT. Jonn Е. SHEPLEY. ALBERT T. PERKINS. Евер С. ZEIBIG. EX-OFFICIO MEMBERS: , OR J. Ми, Chancellor of Washington University. pyrex of eet ro of St. Louis. FREDERICK Е. JOHNS GerorGE T. Moor Bishop of the 5” of Missouri. President of ~ Academy of Sci- ence of St. Louis. Автнов А. BLUMEY President of the Board of sica of St. Louis. DANIEL Breck, Secretary. TABLE OF CONTENTS A New Genus of the Acanthaceae.... . И С» е es ek АЛҒЫ Clarence E. Kobuski A Monograph of the American Species of the Genus Dyschoriste..... Clarence E. Kobuski Studies іп Ше Umbelliferae. I... . Mildred E. Mathias Concerning the Status of the Genus 15:01068............- а АИИ David Н. Linder Sources of Energy for Azotobacter, with Special Reference to Fatty Acids....P. L. Gainey The Effect of Ultra-Violet Radiation upon Higher Plants............. Ethel T. Eltinge The Problem of Species in the Northern Blue Flags, Iris versicolor L. and Iris virginica №............ oS REM Edgar Anderson А New Variety of Vernonia Lindheim- ГТ EE Esther L. Larsen Dysosma: A New Genus of Berberida- СВРСИ E Robert E. Woodson, Jr. Studies in the Apocynaceae. Й. | Revision of the Genus Stemmadenia и Robert Е. Woodson, Jr. Studies іп Ше Apocynaceae. III. A Monograph of the Genus Amsonia. . Sr. + oe ИРЕР Жака Robert E. Woodson, Jr. PAGE 9-90 91-108 109-112 113-168 169-240 241—882 333-334 335-340 341-378 Annals of the Missouri Botanical Garden Vol. 15 FEBRUARY, 1928 No. 1 A NEW GENUS OF THE ACANTHACEAE! CLARENCE EMMEREN KOBUSKI Assistant, Arnold Arboretum of Harvard University Formerly Rufus J. Lackland Research Fellow in the Henry Shaw School of Botany of Washington University A critical study of herbarium material of the genus Dyschoriste has revealed a small group of plants which possess sufficient morphological characters differing from Dyschoriste to merit generic recognition. Apassalus? nov. gen. of the Acanthaceae. Calyx profunde 5-fidus. Corolla infundibuliforma; limbus subbilabiatus vel subaequalis; lobi rotundi, convoluti. Stamina 4, didynama, per paria lateraliter contigua vel connata decurrentia; antherae bi- loculares, basi obtusae, non acutae. Stigmatis lobus anticus obliquus vel dilatus, posticus subnullus. Capsula oblongo- linearis. Semina 2-4, plane compressa, suborbicularia.—Herbae perennes. КоПае ovatae, parvae. Flores parvae, solitarii vel in axillis fasciculati. Type species: Apassalus diffusus (Nees) Kobuski. KEY то SPECIES A. Capsule 2-seeded; plants covered with short, hirsute, spreading hairs; QUE PE io о kg A A. diffusus AA. Capsule 4-seeded; plants glabrous vi mm. long, ovate-aubpotund: flowers 8-9 mm. long; КОЛОНИЈА са а, ои ни а 0007 0000 04 А ВВ. Leaves 25-45 mm. long, ovate-elliptic; flowers 11-12 mm. long; (Ah. THE a aeran EN US UI o Vos LOS . humistratus A. diffusus (Nees) Kobuski, n. comb. ELA. 2. Dyschoriste diffusa (Nees) Urb. Symb. Ant. 7: 380. 1912. ! Issued April 30, 1928. * Name dervied from the Greek a, without and хёссаХос, peg, on account of the absence of anther appendages. Ann. Мо. Вот. Garp., Vor. 15, 1928 (1) [ Уот. 15 2 ANNALS OF THE MISSOURI BOTANICAL GARDEN Dipteracanthus diffusus Nees in DC. Prodr. 11: 124. 1847. Dyschoriste humistrata Lindau in Urb. Symb. Ant. 2: 188. 1900, not O. Ktze., namely, as to plants of Santo Domingo. Stems somewhat tetragonal, slender, shortly hirsute, ascending from a perennial base, nodes closely placed, 1-2.5 cm. distant; leaves suborbicular-obovate, broadly obtuse at the apex, nar- rowing into a petiolate cuneate base, shortly hirsute on both surfaces, entire, 10-13 mm. long, 5-9 mm. wide; inflorescence bracteate, axillary; calyx 6-7 mm. long, lobes linear-acuminate, ciliate, 24 total length; corolla white (ex Buch) or pale lilac (ex Tuerckheim), puberulent оп the external surface, 7-8 mm. long, tube extending into a slightly ampliated throat, lobes rounded; anthers didynamous, filaments slightly pilose at the base, anther cells parallel or nearly so, truncate or rounded at the base; ovary 2-celled, glabrous, style linear, pubescent a little above the base, stigma dilated, oblique; capsule 6-7 mm. long, 2-celled, each cell containing a single seed attached by the retinaculum, both of which (retinacula) are situated on the central ridge of the com- missural surfaces; seeds flat, orbicular, becoming mucilaginous when wetted. Distribution: Islands of Haiti and Santo Domingo. Specimens examined: Haiti: on rocky outcrop, dry wooded mountain slope, vicinity of St. Marc, 25-28 Feb. 1920, Е. С. Leonard 2913 (US, G); dry bank along road near Ennery, Dept. of Artibonite, 325-900 m. alt., 13 Jan. 1926, Е. С. Leonard 8823 (US); arid thickets, north- east of the N. West Indies Company, vicinity of St. Michel de l'Atalaye, Dept. du Nord, 300 m. alt., 17 Хоу. 1925, Е. С. Leonard 7093 (US); common in dry thickets, vicinity of 56. Michel de l'Atalaye, Dept. du Nord, 350 m. alt., 26 Nov. 1925, E. C. Leonard 7472 (US); Barahona, 1200 m. alt., Sept. 1911, Fuertes 1407b (FM, G, US). Santo Domingo: Azua, March, 1913, Rose, Fitch & Russel 4072 (05). A. cubensis (Urb.) Kobuski, п. comb. PLE Dyschoriste cubensis Urb. Symb. Ant. 7: 381. 191. Dyschoriste humistrata Lindau in Urb. Symb. Ant. 2:188. 1900, not O. Ktze., namely, as to plants of Cuba. 1928] KOBUSKI—-A NEW GENUS ОЕ ACANTHACEAE 3 Ruellia diffusa Grisebach, Cat. Pl. Cub. 195. 1866 (excl. syn.); балу. Fl. Cub. 97 (no. 1500). 1873. Low-growing perennial, decumbent, occasionally rising erect, glabrous or minutely scabrous, young stems densely covered with cystoliths; leaves shortly petiolate, ovate to suborbicular, 9-12 mm. long, 5-7 mm. wide, rotund at the apex, tapering to a cuneate base, entire, densely covered with cystoliths on both surfaces, glabrous; flowers solitary, rarely in twos, bracts narrowly obovate; calyx 5-cleft, 6-8 mm. long, lobes linear-acuminate, nearly 24 total length, entire external surface covered with cystoliths, glabrous, lobes ciliated; corolla 8-9 mm. long, tube cylindrical, enlarging until ampliated throat is reached, lobes shortly obovate; stamens didynamous, adnate to the middle of the tube, anthers narrowly ovate, obtuse at the base; ovary 2-celled, style linear, nearly glabrous; capsule oblong-linear, 7-8 mm. long, glabrous, 4-seeded; seeds suborbicular, mucilaginous when wetted. Distribution: near Cojimar, Prov. of Havana, Cuba. Specimens examined: Cuba: near Cojimar, Prov. of Havana, 14 March, 1906, Baker 2894 (FM); shady places in coastal sand between Rio Cojimar and Playa de Bacuranao, Prov. of Havana, 26 Dec. 1910, Wilson 9533 (G, US). A. humistratus (Michx.) Kobuski, n. comb. EL Dyschoriste humistrata (Michx.) O. Ktze. Rev. Gen. Pl. 2: 486. 1891. Ruellia humistrata Michx. Fl. Bor.-Am. 2: 23. 1803; Pursh, Fl. Am. Sept. 2: 421. 1814. Calophanes humistrata Shuttleworth ex. Nees in DC. Prodr. 11: 108. 1847; Gray, Syn. Fl. N. Am. ed. 1, 2: 324. 1878, and ed. 2. 1886; Chapman, Fl. Southeastern U.S. ed. 1, 1083. 1860, and ed. 2. 1889. Dipteracanthus humistratus о. Fl. Southeastern U.S. ed. 2, 303. 1889. Dipteracanthia riparius Chapman, Fl. Southeastern 0.5. ed. 2, 303. 1889. Stems several, ascending or rising erect from a ligneous peren- nial base, 4 dm. or less high, glabrous or slightly pubescent; [Vor. 15 4 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN leaves ovate-elliptic to oblong-sublanceolate, 2.5-4.5 cm. long, 1-2 em. broad, obtuse to acute at the apex, abruptly attenuated at the base into a petiole which may be so short as to give the leaf a subsessile appearance or as much as 4 mm. long, glabrous or nearly so, entire or slightly crenulate margins; bracts oblong- oblanceolate, about equalling the length of the flower; flowers axillary; calyx deeply 5-parted, 9-10 mm. long, glabrous or slightly pubescent, lobes subulate-setaceous; corolla small, white, 10-11 mm. long, tube 2.5-4 mm. long; stamens didynamous (very seldom 5), filaments pubescent at point of adnation to corolla throat, anther cells obtuse or slightly mucronulate at the base; mature capsule 9-10 mm. long, glabrous, linear, 4-seeded. Distribution: low grounds, southeastern United States. Specimens examined: Georgia: Lumber City on the Ocmulgee River, Telfair Co., July, 1900, C. Mohr (US, 721392); shaded places in Ogeechee River swamp, Burke Co., 5 June, 1901, R. M. Harper 769 (M, US). Florida: fertile ground under oaks, upper St. John’s River, 1 June, A. H. Curtiss 28 (а); Hot Springs, 7 April, 1925, H. O’Neill 601 (US); Pine Island, St. John’s River, 11 April, 1911, S. C. Hood (G); swampy shore of St. John’s River, June, 1878, A. H. Curtiss 1939 (M, FM, G, US); wooded banks of the Suwan- nee River at Branford, Suwannee Co., 9 June, 1900, А. Н. Curtiss 6654, (С, M); Suwannee Co., June-July, 1898, А. S. Hitchcock 1457, 1468 (FM); damp shady places, banks of Rice Creek, Putnam Co., 26 March, 1882, C. Mohr (US 721391); Dunnellon, Marion Co., 25 Feb. 1891, L. F. & R. Ward (US, 147428); Port Orange, Volusia Co., 20 Мау, 1895, F. C. Straub 164 (а); Lake Alfred, Polk Co., 11 June, 1922, С. М. & J. К. Armstrong (М 911680); swamp, Hernando Co., June-July, 1898, А. S. Hitchcock (M 120820). озю; огл фет ГУ оз. 15, 1928] ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 1 Apassalus diffusus (Nees) Kobuski Open calyx. Open corolla showing stamens. Dehiscing capsule showing seeds and retinacula. Apassalus cubensis (Urban) Kobuski Open calyx. Open corolla showing stamens. istil. Dehiscing capsule showing seeds and retinacula. Apassalus humistratus (Michx.) Kobuski Open calyx. Open corolla showing stamens. Dehiscing capsule showing seeds and retinacula. ANN. Мо. Вот. Garp., Vor. 15, 1928 PLATE 1 KOBUSKI—A NEW GENUS ОЕ ACANTHACEAE [Vor. 15, 1928] 8 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 2 Fig. 1. Apassalus diffusus (Nees) Kobuski From the specimen, Fuertes 1407b, in the United States National Herbarium, Fig. 2. Apassalus cubensis (Urban) Kobuski From the specimen Baker 2894, in the Herbarium of the Field Museum. AVAOVHINVOV ЯО SONHD MAN У—ТУИЗПЧОМ Ann. Mo. Bor. GARD., Мог. 15, 1928 PLATE 2 A MONOGRAPH OF THE AMERICAN SPECIES OF THE GENUS DYSCHORISTE! CLARENCE EMMEREN KOBUSKI Assistant, Arnold Arboretum of Harvard Universit Formerly Rufus J. Lackland Research Fellow in the Henry Shaw School of Botany of Washington University INTRODUCTION Any attempt to determine specifically herbarium specimens of the genus under present consideration formerly proved very unsatisfactory because of the inadequacy of many of the original descriptions and also because of the small representation of type or authentic material in American herbaria. These facts, along with an especial interest in the genus and its allies, led to the present study. At first it was hoped that a monographic treat- ment of the whole genus might be made. A general survey of the material deposited in American herbaria, however, showed the Old World species to be so poorly represented that it was deemed advisable to exclude them from the present discussion and to include only the American species. Later the writer plans to visit some of the larger European herbaria and to supplement this monograph by a critical study of the far-eastern species. This investigation was made possible only through the собрег- ation of the botanists connected with the various herbaria from which material was borrowed. Sincere appreciation is due Dr. B. L. Robinson of the Gray Herbarium, W. R. Maxon of the United States National Herbarium, and D. C. Davies, Director of the Field Museum, who so willingly loaned their entire col- lections of Dyschoriste for this study. It was found necessary also to borrow types, and to obtain fragments and photographs of type collections from several European herbaria. Dr. Santiago Ramón y Cajal, Instituto Cajal, Madrid, and Professor Eduardo Balguerias y Quesada, Jardin Botanico, Universidad de Madrid, 1 Ап investigation carried out at the Missouri Botanical Garden in the Graduate Laboratory of the Henry Shaw School of Botany of Washington University, and submitted as a thesis in partial fulfillment of the requirements for the degree of doctor of philosophy in the Henry Shaw School of Botany of Washington University. Issued April 30, 1928. ANN. Mo. Вот. Слво., Vor. 15, 1928 3 (9) [Vor. 15 10 ANNALS OF THE MISSOURI BOTANICAL GARDEN very kindly furnished an excellent photograph of a little-known species, the type of which is preserved in the Madrid Herbarium. Professor Boris Fedtschenko, Jardin Botanique Principal, Lenin- grad, U.S.S.R., obligingly supplied two types essential for the completion of thismonograph. Dr. A. W. Hilland T. A. Sprague, Royal Botanic Gardens, Kew, Dr. L. Diels, Botanischer Garten und Museum zu Berlin-Dahlem, Dr. C. H. Ostenfeld and Dr. Carl Christensen, Botanisk Garten, Kgbenhavns Universitet, as well as Dr. Adele Lewis Grant, Huguenot College, South Africa, who so willingly made critical comparisons with types at the Kew Her- barium on her journey to Africa, have all contributed either directly or indirectly, in material loaned or in verification of specimens submitted for comparison. The writer takes this op- portunity to express his gratitude for their generosity and kindly assistance. This study was made at the Missouri Botanical Garden, and thanks are due to the Director, Dr. George T. Moore, for the use of the excellent library and herbarium facilities which this in- stitution affords. Also, especial thanks are extended to the Curator of the Herbarium, Dr. Jesse M. Greenman, under whose constant guidance and supervision this work has been made possible. HISTORY The genus Dyschoriste was proposed by Nees in the third volume of Wallich's ‘Plantae Asiaticae Rariores’ 1 published in 1832. The genus was segregated from Ruellia on account of stamen, corolla, and fruit characters, and was based on Ruellia depressa Wallich, namely, Wallich’s No. 2379 from East India, which, however, is not conspecific with Ruellia depressa L. Two other East Indian species, D. cernua and D. litoralis, were referred by Nees in the same work to his new genus. In 1833, only a year later, David Don in Sweet’s ‘British Flower Garden’? described the genus Calophanes. Don’s new genus was also a segregate from Ruellia, and was founded on Ruellia oblongifolia Michaux, which in turn was based on speci- mens collected in the state of хан ! Wallich, N. РІ. As. Rar. 3: 81. ?Sweet, В. Brit. Fl. Gard. II, 2: x 181. 1833. 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 11 These two generic names were current in botanical literature for many years, as representing two supposedly distinct genera indigenous to remote regions—Dyschoriste of the eastern hemi- sphere and Calophanes of the western hemisphere. Nees, the foremost student in his time of the Acanthaceae, in his treatment of this family for Martius’ ‘Flora Brasiliensis,’ 1 in 1847, accepted Calophanes and described seven Brazilian species of this genus. The same author and in the same year elaborated the Acanthaceae for DeCandolle's ‘Prodromus’ ? and maintained both names as representing separate and distinct genera. In this work, which was the first to present а comprehensive treatment of the group, five species of Dyschoriste and twenty-seven species of Calo- phanes, as well as several varieties, were recognized. Bentham and Hooker in the ‘Genera Plantarum, 2 1876, treated these two previously supposed distinct generic elements as congenerie, but unfortunately they took up the later name Calophanes and relegated Dyschoriste to synonymy. Mr. C. B. Clarke, who contributed the treatment of the Acanthaceae for Hooker's ‘Flora of British India,’ * 1885, fol- lowed Bentham and Hooker’s generic interpretation of Ше group and recognized four East Indian species of Calophanes, namely, C. Nagchana Nees, C. littoralis 'T. Anders., C. vagans Wight, and C. Dalzellii T. Anders. Under C. Nagchana Nees the following species are cited as synonyms: C. depressa T. Anders., Ruellia Nagchana Ham., В. erecta Burm., R. depressa and R. cernua Nees, Dipteracanthus Nagchana Nees, Dyschoriste depressa, and D. cernua Nees. In 1891, Dr. O. Кип ге" revived the name Dyschoriste Nees and transferred thereto several species, including Ruellia erecta Burm. which was described and illustrated in 1768, being the oldest known species of this group. Lindau, in 1895, in reviewing the Acanthaceae for Engler and Prantl’s ‘Die Natürlichen Pflanzenfamilien, * followed Kuntze in recognizing the genus Dyschoriste. 1 Martius, C. F. P. de. Fl. Bras. 9: 26. 1847. 2 DeCandolle, A. P. Prodr. 11: 106, 107. 1847. 3 Bentham, G. & Hooker, J. D. Gen. Pl. 2: 1077. 1876. ‘Hooker, J. D. ЕІ. Brit. India 4: 410. 85. 5 Kuntze, O. Rev. Gen. Pl. 2: 486. 1891. 6 Engler, А. & Prantl, K. Nat. Pflanzenfam. 4%; 302. 1895. (Мог. 15 12 ANNALS OF THE MISSOURI BOTANICAL GARDEN Clarke, in working up the Acanthaceae for the ‘Flora Capen- sis, 1901, took up the name Dyschoriste, thus reversing the position taken by him in Hooker’s ‘Flora of British India,’ mentioned above. He recognized D. depressa Nees as a valid species along with four other African species, one of which is D. erecta Clarke, thus apparently disregarding the D. erecta (Burm.) O. Ktze. Several new species have been described from time to time and referred to either Dyschoriste or Calophanes, but no compre- hensive treatment of the group as a whole has been published since that of Nees in DeCandolle’s ‘Prodromus.’ Assuming that there is absolute identity and thus complete synonymy of the several elements which Clarke referred to Calophanes Nagchana in the ‘Flora of British India,’ then, as pointed out by Dr. Kuntze, the name erecta, as the oldest specific name involved, must be retained and the binomial Dyschoriste erecta (Burm.) O. Ktze. becomes the valid combination for the plant concerned and D. depressa (Wall.) Nees must be regarded as a synonym of it. Since this study has been confined to the American species we must admit the observations of Clarke and Kuntze and accept Dyschoriste erecta (Burm.) O. Ktze. as the type species of the genus until the eastern species in question can be examined. GENERAL MORPHOLOGY Roots.—The root system in the genus Dyschoriste is not very extensive. All the species are perennial and the roots, in turn, are of the simple fibrous form. By the casual observer, however, some of the slender underground stems of the previous year’s growth are sometimes mistaken for roots. Stems.—There is considerable variation in the stem and its habit of growth. In all cases, the stem or stems arise from a ligneous perennial base. However, the mode of ascent varies. Many species are prostrate and the stems spread over the ground in several directions. In these cases the leaves assume a secund position. Often when the stems are short a rosette appearance is attained for the whole plant. The species D. oazacensis illus- ! Flora Capensis 51: 15. 1901. 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 13 trates this character. However, it is not a stable specific char- acteristic. A second habit of growth is the ascending type. It is in this category that the majority of species is placed. The stem not infrequently becomes more or less geniculate; this mode of growth is very characteristic of D. Pringlei. The third habit of growth is the erect type. It is to this type that the sturdy D. hirsutissima, D. oblongifolia, D. ovata, and D. trichanthera belong. Some species may follow consistently a distinct habit of growth, while others may have the stems ascending or erect even on the same plant. One can usually associate stem growth exceeding a length of 4-5.5 dm. with the erect habit, and shorter stems with the ascending or prostrate habit. Along with this, the prostrate type will possess small leaves and the erect type will have leaves with a more extensive surface. Several species, among them JD. oblongifolia, possess slender underground stems which are common in perennials. These stems have the appearance of roots but on close observation buds and modified leaves can be seen. After passing under the ground for some distance they come to the surface and then rise erect. The stem may be terete or quadrangular. The latter is the more common type іп the genus. Іп the species D. quadrangularis the stem is not only angular but the angles are winged. This condition is probably brought about by the decurrent petiole of the leaf extending down the stem. Leaves.—The leaves of the species in the genus Dyschoriste present a great variety of differences. АП species have leaves with entire margins, except D. bilabiata which is not distinctly dentate but has a decided tendency in that direction. Several species, such as D. crenulata and D. hirsutissima, show a tendency toward crenulate margins. Others combine the crenulate with the repand margin. Along with these characteristics the margin is usually ciliate. In shape the leaf varies from that of the narrow, linear D. angusta, D. Greenmanii, and D. Purpusii to that of the oblong-ovate D. quadrangularis which grows to a length of 10 centimeters. Some species have two types of leaves, the lower or cauline leaves often being larger and different in shape from the upper leaves in whose axils branches and flowers are crowded. (Хот, 15 14 ANNALS OF THE MISSOURI BOTANICAL GARDEN The surface of the leaf itself is usually pubescent. When the pubescence is sparse or absent, an abundance of cystoliths is usually seen. Often the cystoliths, since they lack an orderly arrangement, are mistaken for appressed hairs. This cystolithic character is not sufficiently definite for specific delimitation. Some plants have a more copious formation of cystoliths than others. This character exists on the stem and calyx as well as the leaves. The venation which is very pronounced on the lower surface varies very little within the genus. The usual form is the feather-veined type. Inflorescence.—In all cases the flowers are axillary and sub- tended by bracts and sometimes bracteoles. Only occasionally are the flowers solitary in the axils. "There are usually several to many flowers at the node, giving the appearance of a cymose cluster as in D. quadrangularis or а capitulum as in D. capitata and D. pinetorum. D. Greenmanii is an excellent example of a species with a solitary flower at the node. The majority of species have flowers which are pedicellate, often so short, however, that a subsessile effect is presented. Calyx.—In the calyx of Dyschoriste is found one of the most constant characters of the genus. It is usually five-parted and always persistent. The only deviation from the five-merous condition is found in D. maranhonis where the calyx-lobes are occasionally only four. In all cases the lobes are subulate- setaceous and usually ciliate. The ciliation may vary from a long whitish, flaccid pubescence to a very short hirsuteness. When the calyx proper is pubescent, the pubescence is usually confined to the nerves. The tissue connecting the lobes of the calyx is usually very membranaceous and tears apart very easily, making it difficult and quite unsatisfactory to use the ratio between tube-length and lobe-length as a character for specific differentiation. The lobes are usually quite equal in length. However, here again variation is found. Cystoliths, as in the leaves, are very abundant; but in the calyx they are frequently disposed in a more or less regular arrangement. Corolla.—There is very little differentiation to be found in the corolla of the genus. In most cases the bilabiate type occurs. This, however, is not as distinct as in some other genera of the 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 15 Acanthaceae. The corolla is five-lobed, the two posterior lobes being coalescent to a greater extent than the three anterior lobes. The length of the corolla varies from 10 to 17 mm., as found in D. decumbens, D. hygrophiloides, D. saltuensis, D. quadrangularis, and D. angustata, to 25-28 mm., exemplified in D. zylopoda, D. humilis, and D. ovata. One species, D. Pringlei, has а corolla measuring 35-38 millimeters long. None, however, reach the length of 70-80 millimeters as found in some of the Ruellias. The proportion between tube and throat is variable in the genus. The ventricose throat is found quite often. The condition should exist in all species because of the contiguity of the adnate fila- ments in the posterior portion of the throat and tube. This ventricosity, hence, is more pronounced in the larger-flowered species. The narrow tube of the corolla is usually slightly flared at the base to make room for the disc and ovary. The ampliation from the tube to the throat is very variable and may be abrupt or gradual according to the species. In all cases, the external surface is quite pubescent. In the species D. trichanthera the pubescence is found on the interior as well as the exterior surface of the throat. Stamens.—The stamens are didynamous. The long and short filaments on each side are contiguous or united at the base by a membrane which extends from the point of adnation to the base of the corolla tube. A very distinctive feature of the anthers is the mucronate appendages at the base of the anther cells. These mucronate appendages are characteristic of the genus and are very easily seen with the hand-lens. Under the low power of the compound microscope they are found to be composed of several multicellular strands of cells closely compacted together. These strands of cells are easily torn apart, and a dentate or ragged appearance is given to the whole appendage. ‘This is doubtless what Nees saw when he described the appendages of D. quitensis as 2-3-toothed." А similar example was found in D. Schiedeana. On microscopic study, however, the so-called dentations were found to be nothing more than shreds of tissue torn away slightly from the compacted mass. The anther cells are usually parallel and oblong in shape. In the case of D. sagit- tata and D. maranhonis the cells are so disposed as to have a [Vor. 15 16 ANNALS OF THE MISSOURI BOTANICAL GARDEN sagittate appearance. In both the species mentioned the apex as well as the base is appendaged. Аз a rule the anther cells are glabrous but in the species D. trichanthera the anther cells are very pubescent. The mode of dehiscence is by a longitudinal slit on the side of the anther cell. The filaments are commonly pubescent. Pistil.—There is little variation in the parts of the pistil. A disc is present beneath the ovary in all species. The ovary itself is two-celled, glabrous, and oblong. Little or no variation is found in the filiform pubescent style. Only the anterior lobe of the stigma is developed, and this lobe is usually linear and oblique with a flattened stigmatic surface. However, in D. hygrophiloides the stigma is curved, while in D. sagittata it is basally lobed. In D. maranhonis the stigma is reflexed. Сарвше.--Тһе capsule of the genus is quite uniform. The constant linear, glabrous and four-seeded characters, combined with other diagnostic characters, help considerably in generic determination. Retinacula or hooked appendages on the median ridge of the valves hold the flat suborbicular seeds in place. When dry the seeds appear to have many soft, appressed hairs. These same hairs when wetted diverge, elongate, and become mucilaginous. GEOGRAPHICAL DISTRIBUTION The geographical distribution of the American species of the genus Dyschoriste offers very interesting problems. The accom- panying maps demonstrate very clearly that there are three distinct areas of distribution: (1) southeastern United States; (2) southwestern United States and Mexico ; and (3) South America. Two species, namely, Dyschoriste oblongifolia and D. angusta, occur in the southeastern United States area. This area extends the width of the coastal plain from southern Virginia to southern Florida. The regions of distribution of the two species do not overlap. D. angusta is confined to the wet region of Dade and Palm Beach Counties in southern Florida, while D. oblongifolia extends northward through the remainder of the area, seeking the dry, sandy pine woods. 1928] KOBUSKI—MONOGRAPH ОҒ DYSCHORISTE 17 ` M v їе | В % E . ^ “4 af “.. в X OE \ ( 7 з i - 1 ‘ ` тепкен => E a p^)». Ч а K ( | SALA : S MP У (c ау — A \ ! г ! ~ (4 5 р Н : 3 ча 145 4 Ири Хо / М Фа 3 ' : @ > “У ( р ~ ба МУ NN р ~ 3 5 5 СҮҮ 1 / \ 1 i я 1 1 / Aa А ; p Y 4 9” | | 1 i 4 4 MAI 2 7) d ! | : | р — Ф 4 еуі КАЗАК |Т АИК ЛАН 77707777 - 2 i ! > t 8 эе MR | A ОМ if Ae ea 1 | Бе , | H | | " x А of ПИ FT ’ ч 15 M И і } мді “> 2 7 КУ Ч Ни е - і 27 Е J ' | hi 5 s E | ~ | p , р КЕЙИ 8-1 Жл ККЕ М E ШТ E apu Pi +a! і | Ж | : E ғ; 5 FER 3 ! ERE 4 Е — 8 а à 8 в ; ч 24k H 3 aff ; ДЕ 3 Ж s / Ч в y i ® Е; х Я ЕЭ З The second area extends from Texas and southern Arizona southward to the Isthmus of Tehuantepec and contains the greater number of species. Of the forty recognized species in the genus Dyschoriste, twenty-one, or more than half, are confined Map showing the geographical distribution of the North American species of Dyschoriste. Fig. 1. 18 Vor. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN os Т he K = ~ 2) а %%; = Ка 4 9 1) : р ; 1 ww Т9 зора Боэчоненюы КЕКЕКХЕРЕКККІ ЖЕЛ SARL |- № зра. 38 НЕ H cm 7» Б 722444 - СЗО Ира: Fig. 2. Мар showing the geographical distribution of Ше South American species of Dyschoriste. to this area. It is an interesting fact that the distribution of some species is almost coincident with the geological formation of the country. D. decumbens, which occurs on the plateau 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 19 region between the Sierra Madre ranges, is an excellent example of this fact. D. hirsutissima extends from Sonora southward along the western slope of the Sierra Madre range to Oaxaca. Many species have a localized distribution only, D. Greenmanit, D. crenulata, D. saltuensis, and D. angustifolia being examples. Some of these localized areas are characterized by three or more species. Ап instance of this is а small region around Guadalajara in the state of Jalisco where four species are represented. Ап- other illustration of limited areal distribution occurs in the northern part of Oaxaca which harbors D. oaxacensis, D. angusti- folia, D. capitata, and D. hirsutissima. Many species, especially the localized ones, appear to be extremely едар с since they inhabit only regions near volcanoes. Тһе center of distribution in this second area falls within the region represented by the states of Puebla, Michoacan, and Mexico. The third and last area, namely that of South America, com- prises more territory than either of the areas indicated above and includes the seventeen remaining species of the genus. Тће material examined in all cases was not very copious, hence an accurate range of geographical distribution of these species could not definitely be ascertained. Nearly all species appear to occur in isolated and limited areas, but the relationship between some of these areas indicates that a greater overlapping of areas would occur were it not for the paucity of herbarium material. Three species, namely, D. quitensis, D. ciliata, and D. repens, are found in Peru and the Andes of Ecuador. D. Niederleinii and D. humilis are found in Argentina. The other thirteen species inhabit northern Paraguay and southern Brazil, and it is here that the South American center of distribution occurs. An unusual feature of the geographical distribution is the isolation of the areas defined. At present, there is no one species which connects up any two areas. There seems to be no satis- factory explanation to account for the absence of the genus between the Andes of Ecuador and the Isthmus of Tehuantepec in Mexico. Members of the genus may be found between these two remote regions, but until the entire Andean range has been explored more thoroughly from a botanical standpoint one would hardly venture an explanation of the marked discontinuous dis- tribution which the genus now presents. (Vou. 15 20 ANNALS OF THE MISSOURI BOTANICAL GARDEN The non-occurrence of the genus in the Mississippi Valley is equally surprising. Since the flora of this entire area is com- paratively well known, it is hardly possible that the members of the genus would be overlooked if they there existed. The only solution seems to be the possible age of the genus. Dyschoriste is probably a pre-glacial genus which, prior to the Oligocene period of the Cenozoic era, extended continuously across the southern United States. However, the Eocene and Oligocene seas encroached upon the United States in the present Mississippi Valley, thereby splitting the distribution areas of the genus into two parts. PHYLOGENY Because of the large number of closely allied species in the genus Dyschoriste it is quite necessary that the phylogenetic discussion of the group be made from a purely hypothetical standpoint. The fact that the discussion is confined to the American species alone seconds this consideration, since the eastern species of the genus exceed the American species in number. On account of the three distinct geographical distribution areas, which have been discussed before, a tree method of illus- trating probable phylogenetic sequence proved unsatisfactory; hence the method used in the accompanying chart was devised finally to illustrate the apparent relationship of the species of the western hemisphere. This chart if superimposed on a map of the regions inhabited by the genus would coincide with the specific regional distribution. It was felt reasonably certain that all species of the genus have evolved from a common ancestor designated in the chart as x From this ancestor, species and groups of species have evolved. One might ask why, since the species seem to be placed in definite groups, subgenera or sections have not been designated. This question was given much thought and consideration; it was felt finally, however, that on account of the relative uniformity of the essential morphological characters within the genus, except in the case of group II, no adequate basis exists for the designation of subgenera or sections It may be observed that all the designated groups with the 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 21 exception of group II originated from the common ancestor at approximately the same time. Group П, on account of the muticous character of its anther appendages, the extremely small leaves, the very small flowers, and the fruit characters, has been separated from the genus Dyschoriste and raised to the rank of a new genus which is considered as intermediate between the hypothetical type and Dyschoriste proper. In this genus, Apassalus,! containing three species, the species А passalus diffusus Fig. 3. Phylogenetic chart of the species of Dyschoriste. has reached the highest development in the reduction of the number of ovules to two, one being borne in each valve of the capsule. The other two species contain the four seeds which are characteristic of the majority of species in the genus Dys- choriste. Apassalus is confined to the islands of Cuba and Haiti and to, the southeastern United States. 'See Kobuski, C. E. А new genus of the Acanthaceae. Ann. Mo. Bot. Gard. 15: 1-8, pl. 1-2. 1928. [Уоь. 15 22 ANNALS OF THE MISSOURI BOTANICAL GARDEN Group I, involving ten species, is confined to the plateau regions of Mexico. In this instance D. decumbens, on account of its extended range and characteristic relation to all species con- cerned, is considered the base species. D. Lloydii and D. crenu- lata are species having vital characteristics similar to D. decumbens but differing sufficiently in minor characters to be considered direct descendants from the base species. A small group con- taining three species, D. Purpusii, D. Greenmanii, and D. Rosei, stands by itself. 'The highest development is reached in D. Greenmanii and D. Rosei in which cases the inflorescence has been reduced to a solitary flower at each node. АП species of the last group have very slender linear leaves. Another branch from D. decumbens, as the chart illustrates, is the linearis-jaliscensis branch. Although separated somewhat in regional distribution these two species are closely allied through their flower and foliage structures and are undoubtedly derived from D. decumbens. The species D. oblongifolia and D. angusta seem to be closely related to D. linearis; in fact, D. linearis was once considered а variety of D. oblongifolia. However, the two species under dis- cussion are confined to the southeastern United States in their distribution and, as the distribution map of North America shows, are not connected definitely with the Mexican-southwestern United States species. This suggests the probability that there may be a relationship between the two species in Florida and the genus Apassalus also found there and in the West Indies. The migration may have been northward through the Antilles, and a connecting link between the southeastern United States species and the southwestern United States-Mexico group may never have existed. In group III, one finds an entirely different situation. Here we have four large, erect species, each showing a definite charac- teristic development toward advancement. Perhaps the highest development is found in D. hirsutissima which extends the length of the western Sierra Madre range and possesses а well-developed glandular pubescence. This is the only instance of this character in the whole genus. D. bilabiata and D. quadrangularis are close relatives but do not seem to have been derived from the D. hir- 1928] KOBUSKI—MONOGRAPH ОҒ DYSCHORISTE 23 sutissima line. Instead, they undoubtedly arose along parallel lines of development. In D. bilabiata we find a distinct dentation of the leaf. This group is also characterized by large, petiolate leaves, in some cases as long as ten centimeters, an unusual feature in the genus. Group IV for the most part occurs in southwest Mexico, that is, the states of Oaxaca, Jalisco, and Michoacan. These species are the ascending foliose type with rather small, ovate leaves. The inflorescence is usually subcapitate, and it is sometimes dif- ficult to distinguish off-hand the species here included. They all, with the exception of D. pinetorum, seem to have evolved along a parallel line of development. D. pinetorum, on account of its resemblance to D. Риме, undoubtedly evolved from it. It is in D. Pringlei that we find the largest flower of the genus Dyschoriste, and in this species a close resemblance is shown to the genus Ruellia. The highest point of development in the North American species just discussed is found, according to my opinion, in D. hirsutissima, D. Greenmanii, D. Rosei, and D. linearis. In the South American species of the genus, a similar situation is found. Неге the species can be placed in five groups. Group V contains the three species on the western coast inhabiting Peru and Ecuador. Specialization in them is not particularly noticeable. The species D. ciliata possesses rather muticous anther appendages. This would ally the species to the genus Apassalus. It was necessary to accept the word of Nees in this instance, however, as only a photograph of the type of the species could be obtained. The species of Group VI are found in Argentina. Неге D. humilis reaches the highest point of development in the reduced number of seeds. As in Apassalus diffusus, only two seeds are produced, a single seed occurring in each valve of the capsule. Group VII is not unusual in its development. In this group are found six closely related species—perhaps interrelated— but showing no special development. Here again the herbarium material at hand is very sparse, the study in a few instances being confined to photographs. In Group VIII are four species, two of which are described for [Vor. 15 24 ANNALS OF THE MISSOURI BOTANICAL GARDEN the first time. In the species D. trichanthera is found the spicate inflorescence along with pubescent anthers. The former char- acter links it up with D. Schottiana, while the latter character shows the relationship which exists between D. trichanthera and D. lavandulacea. D. paraguariensis is undoubtedly a branch from D. lavandulacea but possesses more highly developed floral characters. The last group, namely, group IX, includes only two species. It is here that the sagittate or divergent anther cells are found. The highest development of the group and probably the highest development in the genus is shown by D. maranhonis, in which both incomplete didynamy and reduction in corolla and calyx- lobes are found. Nees describes D. maranhonis as being glandu- lar-pubescent. А fragment of the type specimen was obtained and failed to demonstrate this character. SUMMARY The conclusions drawn as to the probable phylogeny of the group under consideration are reached after a comparative study of the outstanding morphological characters which may be sum- marized as follows: (1) Muticous appendaged anthers are more primitive than those with apiculate appendages. (2) Divergent anther cells are more advanced than parallel anther cells. (3) Glabrous anthers are more primitive than pubescent anthers. (4) Four-seeded capsules are more primitive than two-seeded capsules. (5) Numerous flowers in an axil is a more primitive condition than the solitary-flowered axil because the presence of bracts in the solitary-flowered species shows reduction to have taken place in the telescoping of the inflorescence. (6) An unlobed stigma is more advanced than the lobed stigma. (7) Entire-margined leaves are primitive. The dentate margin is an advancement, and the crenulate margin type is intermediate. (8) Glandulosity is more advanced than pilosity. 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 25 (9) Procumbent plants are more primitive than erect plants which have evolved through ascending plants. (10) Complete didynamy is more primitive than incomplete didynamy. (11) Winged stems are more advanced than the unwinged quadrangular stems. (12) Reduction of corolla- and calyx-lobes from five to four is a criterion of specialization and advancement. ABBREVIATIONS The abbreviations used to indicate the herbaria in which the specimens cited in the present paper occur are as follows: В = Botanischer Garten und Botanisches Museum, Berlin, Germany. C = Botanisk Garten, Kgbenhavns Universitet, Copen- hagen, Denmark. Ch = University of Chicago (deposited in the Field Museum). FM - Field Museum of Natural History. С = Gray Herbarium of Harvard University. К = Royal Botanic Gardens, Kew, England. L = Jardin Botanique Principal, Leningrad, U.S.S.R. М = Missouri Botanical Garden. Ma - Jardin Botanico, Universidad de Madrid, Madrid, Spain. US - United States National Herbarium. TAXONOMY Dyschoriste Nees in Wallich, Pl. As. Rar. 3: 75, 78. 1832; Nees in DeCandolle, Prodr. 11: 106. 1847; O. Kuntze, Rev. Gen. Р]. 2: 485. 1891; Lindau in Engl. & Prantl, Nat. Pflanzen- fam. 43°; 302. 1895; Gray, Manual, ed. 7, 743. 1908. Calophanes D. Don in Sweet, Brit. Fl. Gard. 2: pl. 181. 1833; Nees in Mart. Fl. Bras. 9: 25. 1847; Nees in DC. Prodr. 11: 107. 1847; Benth. & Hook. Gen. Pl. 2: 1077. 1873-76; C. В. Clarke in Hooker, Fl. Brit. India 4: 410. 1885; Gray, Syn. Fl. N. Am. 21: 324. 1878, and ed. 2, 1886; Chapman, FI. Southeastern U.S., ed. 3, 365. 1897; Small, Fl. Southeastern U.S. ed. 1, 1082. 1903, and ed. 2, 1913. Vor. 15 26 ANNALS OF THE MISSOURI BOTANICAL GARDEN Linostylis Fenzl. in Linnaea 23: 94. 1850. erbaceous, caulescent perennials, prostrate, ascending or erect, glabrous or pubescent. Leaves opposite, sessile or petioled, usually entire. Inflorescence cymose, capitate or spicate, terminal or axillary. Flowers subtended by foliaceous bracts and bracte- oles. Calyx deeply 5-lobed, lobes usually subulate-setaceous, ciliate, lineolate. Corolla-tube usually erect, occasionally slightly ampliated at the base; limb spreading, oblique, obscurely or distinctly bilabiate, 5-lobed. Stamens 4, didynamous; filaments of a long and short stem united at the base and adnate to the base of corolla-tube, pubescent; anther 2-celled, cells oblong, sharply mucronate at the base, parallel or slightly divergent, glabrous or occasionally pubescent. Ovary 2-celled, glabrous, ovules 2 or occasionally 1 in each cell; style filiform, pubescent; posterior lobe of stigma rudimentary, anterior lobe oblique, slightly flattened. Capsule included in the persistent calyx, oblong-linear, glabrous, 2—4-seeded, separating with difficulty at maturity into 2 valves, 1-2 seeds to each valve held in position by retinacula. Seeds flattened, suborbicular, mucilag- inous when wetted. Type species: Dyschoriste erecta (Burm.) O. Ktze. in Rev. Gen. Pl. 2: 485. 1891. KEY то SPECIES 1. Plants ш апдшаг-риһевсепі.............................. 1. D. hirsutissima Plants glabrous or pubescent but not ріап4ашаг........................... 2 SSO күйді, ок cd ees oh сени ven Peck о ЛЕ 3 т. Ра ИО hae хал Ру 4 3. Anther cells pubescent; internal surface of corolla-throat pubescent. Pleven Gee Nae ра C ORI E PESE La MU RU nee 2. D. trichanthera Anther cells glabrous; internal surface of corolla-throat glabrous.3. D. Schottiana Ж t Па 5 о, onmi Ко ША „сати с пат ess > гэл и 14 5. Corolla approximately 10 mm. Іопр........................... 4. D. angusta Curs I5 cus. oE OI Посл а 6 6 Следа ж.д... ЗОВИ ОО орела кода еи 7 Стоби 20-90 ПИЩИ Ns аена веса Citra CER ы CEA 11 АЕ" ~, Я В. СЕВО ТИИС ти, UTI NS 8 Leaves 1 mm. ог less wide.......... ИЯ 5. Р. Ригризи D „о ОМА ОН. суб та РА 6. D. lavandulacea ДЭГ QUE ВЕ. Порой ce a ее . Stem glabrous, except for distinct pubescence at node; flowers solitary Bb ео РУМЕ 7. D. Стептапи 10 © 1928] KOBUSKI—MONOGRAPH ОЕ DYSCHORISTE 27 10. Plants low-growing, —— 1 dm. or less high; style approximately TuS d: D. ANM. ООВ EMEN 8. D. Niederleinii Plants strict, 3-6 dm. high; style approximately 12 mm. T Mex. sp. ЕРЕ КАРАА КОЧ Е зг... . D. Schiedeana 11. Flowers characteristically — atthe node. .......355. сиви 10. D. Rosei Flowers not solitary at the А er er EX CHER 22 12 12. Leaves mostly linear, 2 mm. or Da ИДЕ. ет 11. D. jaliscensis Leaves linear to linear-lanceolate, more than 3 mm. wide.................. IB Pus IQ DERE. И Е 12. z angustifolia Stem hirsute with rigid hairs. ао я inearis 14. ЕАН dentate.. ec.. eges ez а неки њи E D. bilabiata ХӨВ pot distinctly dentate, ......... ог. 15 15. Cinereous-pubescent #ћгоџцрћоцћ............................ CUR ee ME 16 То а see oc dd ds А сх АКИ 17 16. Leaf margins entire, not crenulate. ....................... 16. D. decumbens Leaf margins distinctly сгепшізде.......................... 16. D. crenulata 17. Inflorescence capitate or виһсарИайе.................................... 18 Inflorescence other than сарбаје...................... CO BORN ERR € 21 18. Anthers emarginate, elongate at the арех..................... 17. D. capitata Anthers without an elongation at the арех.............................. 19 19: Corolla Ви. Jong... ес ер cee 18. D. Pringlei Corola SOME uim. long.......... сура ЭСГИЙ И. 21133. an cee 20 20. Leaves glabrous, објапсеојаће............................ 19. D. oaxacensis Leaves pubescent, ођоуаје-е рне......................... 20. D. pinetorum 21. Corolla 16 im. or lee in length... зада СС. erue 22 Corolla 2 ЗАЛ, ог more in length. ......./ ХОМС Gs ро оек 33 а RERO оо с... 23 Leaves avo ао oo 070 а а рано 25 23. Plants with converging anther cells giving a sagittate appearance; stigma ЮБей...................Ж. тко с НП. г 21. D. sagittata Plants with parallel anther cells; stigma ишођеа......................... 24 24, Plants with glabrous leaves; S. Amer. вр................... 22. D. Serpyllum Plants with pubescent leaves; Мех. вр........................ 23. D. ГЛоудњ 36. Тайна Майко. ..........- д 26 Leaves: pullment.......... 4 352922 о, conan 28 20. Tes 8-7 em. сәз... К AU eden окна 27 Losyen 1.5-8 em. long. eneee хот e ib ss 24. D. microphylla 27. Anthers distinctly calearate at base; leaves lanceolate; Mexican species. Per Ter а бозони невин aas M о. 22298 25. D. saltuensis жаза 824 slightly calcarate at base; leaves ovate-elliptic; В. хярс BDecleB. рака ок ЛИН ИЕ . D. ciliata 28. Stems ям leaves approximately 10 cm. long........ 27. D. Сред ари Stems not winged; leaves 2-4 сіп. Іопр.................................. 29 29. Calyx-tube glabrous, lobes ciliate with softly hirsute hairs; leaves lanceo- 816...» рала ee ns Е с. 28. D. quitensis ET мон pubescent, lobes ciliate with stiffish hairs; leaves ovate to : шы cells diverging, giving а sagittate appearance; corolla and calyx lobes frequently reduced to 4; incomplete didynamy co О Аы. 29. D. maranhonis е © (Мог. 15 28 ANNALS OF THE MISSOURI BOTANICAL GARDEN Anther sm parallel; corolla and calyx characters constant; complete didyn PANNI. соки auTO КСЛ rd E ee eee 31. Style шини 5 mm. 2 О у ~ | сы 32 owen ЛО CY Po „не. рт ~ АИК 80. D. hirsuta 32. Lower leaves obovate to d. emarginate at the apex В А 81. D. hygrophiloides Lower leaves 2 obtuse but not emarginate at Ше apex...... 32. D. repens бео lO i Pe РЕКА Tere err est ИРУ 83. р. Pulegium Calyx 15 mm. or мағы Л ПИРИ Т ЛГ (| 34 А __- 7 ·_. о |" И 35 | _ аду in pairs аб М nodes. .....››..... “= = > 36 35. Stem erect; corolla barely exceeding or equalling the calyx in а ааа ғ D. ovata Stem geniculate-escending; corolla 5 mm. or more longer than edi бүл М “МЭТ И Т ОГ ОЛ o. . D. amoena 5" п. ААУ 7" жк иа крача Кел > 7 МИН ЛЭ... гал гу ИЕ 89 37. Villous-pubescent іһтоц һош.............................. 36. D. zylopoda и =. И ERE 38 38. Capsule 4-seeded; М. Amer. вр........................... 87. D. oblongifolia сик кімі; В. Amer. брег с: oec хха аа ажээ 38. D. humilis 39. Leaves distinctly acuminate; the two bracts at each axil реци equalling __- B 1 Ro cor Зөрүү лэ err re . D. paraguariensis Leaves obtuse at apex; bracts foliate but not equalling the и d РА 40 40. Leaves sessile; corolla 25-28 mm. long; Am. bor. sp. re er rer vid 9t asd te 37а. D. oblongifolia f. glabra Leaves petiolate; corolla 20 mm. long; 5. Amer. sp.......... 40. D. Tweediana 2 1. Dyschoriste hirsutissima (Nees) О. Ktze. Rev. Gen. РІ. 486. 1891. Calophanes hirsutissimus Nees in DC. Prodr. 11: 109. 1847; Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. Calophanes bilobatus Rose in Contr. U.S. Nat. Herb. 1: 109. 1891 Stems branching, ascending from a stout perennial base to a height of 10-12 dm., somewhat quadrangular, more or less pubescent with the pubescence, in some cases, restricted to the edges, occasionally glandular at the apex; leaves petioled, ovate to oblong-ovate, 3-8 cm. long, 1.5-3 cm. wide, acute at both ends, margin usually entire or slightly crenulate, sometimes slightly denticulate, pubescent on both surfaces, younger leaves often densely so and glandular; inflorescence axillary, subtended by glandular-pubescent, subulate bracts; calyx 5-lobed, subulate- setaceous, extremely glandular-pubescent, approximately 11 mm. 1928] 5 KOBUSKI—MONOGRAPH OF DYSCHORISTE 29 long, lobes more than one-half the total length of the calyx; corolla subbilabiate, puberulent on the outer surface, occasionally glandular, averaging 14 mm. in length, tube about the same length as the abruptly ampliated throat; stamens adnate to a little below the middle of the corolla-throat; stigma oblique; capsule 4-seeded; seeds облаче. Distribution: slopes of the Sierra Madre of western and southern Mexico. Specimens examined: Southwestern Chihuahua, Aug.-Nov. 1885, Palmer 285 (С, US); Alamos, 180 miles з.е. from Guaymas, Sonora, alt. 418 m., 26 March-8 April, 1890, Palmer 402 (G, US); Sierra de los Alamos Mt., 6 miles due south of town of Alamos, Sonora, 14 March, 1910, Rose, Standley & Russell 12833 (US); dry hillside, Acaponeta, Теріс, 10 Apri., 1910, Rose, Standley & Russell 14298 (US); dry rocky slopes nesr Guadalajara, Jalisco, 11 Dec. 1888, Pringle 2154 (G); hills near Guadalajara, Jalisco, 15 Nov. 1889, Pringle 2939 (FM, G); Cuernavaca, Morelos, 15 Nov. 1865, Bourgeau 1262 (G); Cuicatlan, Oaxaca, alt. 1000 m., 9 Dec. 1895, Gonzalez 43 (G); Monte Alban, Oaxaca, 1933 m., 23 Nov. 1894, Pringle 6053 (G, М, US); Monte Alban, Oaxaca, alt. 2833 m., 24 Nov. 1894, L. С. Smith 328 (С); Monte Alban, near Oaxaca City, Oaxaca, alt. 1900-2000 m., 23 Nov. 1894, L. С. Smith 729 (M, US); Monte Alban, Caxaca, 29 Dec. 1895, Seler 1733 (G); Valle de Oaxaca, Oaxaca, alt. 1650 m., 18 Nov. 1906, Conzatti 1521 (FM); Tehuantepec, June, 1906, Gandoger (М 120892); Hacienda de Guadalupe, date lacking, Ehrenberg 1223 (B TYPE, М fragment and photograph). 2. Dyschoriste trichanthera' n. sp. Pl. 4. Dyschoriste maranhonis Lindau in Bull. Herb. Boiss. П. 3: 628. 1903, as to Fiebrig 4856, Hassler 5908, 7780, not O. Ktze. Stems stout, branched, erect, 5-6 dm. high, glabrate, pubescent near the apex, basal portion densely covered with cystoliths, swollen at the nodes; leaves oblong-ovate to ovate, younger 1 Dyschoriste trichanthera Kob., sp. nov., caulibus suffruticosis, erectis, 5-6 dm. altis, glabrescentibus, apice pubescentibus, inferiore cystolitherissimo, tumidis ad nodos; foliis oblongo-ovatis vel ovatis, 5-7 cm. longis, 2-3 cm. latis, integerrimis vel crenulatis, petiolis 10-12 mm. long s; floribus axillaribus, aliquot ad singularos nodos, fere prope apicem spicatis; bracteis parvis, foliaceis, ciliatis, pubescentibus, bracteolis acuminatis, 4-7 mm. longis; calyce 13-14 mm. longo, lobis subulatis, setaceis, 8-9 Vor. 15 30 ANNALS OF THE MISSOURI BOTANICAL GARDEN leaves pubescent, older leaves glabrate, 5-7 cm. long not including the petiole, 2-3 em. wide, entire to crenulate, petiole 10-12 mm. long; flowers axillary, crowded at the nodes near the apex giving a spicate appearance; bracts small, foliaceous, ciliate and pubes- cent, bracteoles acuminate, 4-7 mm. long; calyx 13-14 mm. long, lobes subulate-setaceous, 8-9 mm. long, often recurved at the tip, ciliate, with 2 kinds of multicellular hairs, both flaccid and delicate; corolla distinctly bilabiate, 10-20 mm. long, rose or violet, lobes obtuse, emarginate, puberulent on external surface, distinctly pubescent on the internal surface; stamens barely included, adnate to about opposite the labiation of the corolla, anthers pubescent, style filiform, 10-20 mm. long, stigma oblique, linear; capsule not seen. Distribution: along rivers, northern Paraguay. Specimens examined: In the region of the river Capivary, Paraguay, date lacking, Hassler 5908 (G); between the rivers Apa and Aquidaban, Paraguay, Jan. 1908-9, Fiebrig 4856 (С); in region along the river Apa, Paraguay, Nov. 1901, Hassler 7780 (С түре, M photograph and fragment). 3. Dyschoriste Schottiana (Nees) Kobuski, n. comb. Hygrophila Schottiana Nees in Mart. Fl. Bras. 9: 22. 1847; Nees in DC. Prodr. 11: 87. 1847. Dyschoriste crinita (Nees) О. Ktze. Rev. Gen. Pl. 2: 485. 1891; Lindau in Bull. Herb. Boiss. 7: 575. 1899. Calophanes crinitus Nees in Mart. Fl. Bras. 9: 26. 1847; Nees in DC. Prodr. 11: 107. 1847. Herbaceous perennial; stem erect, 5-6 dm. high, profusely branched, hirsute; leaves oblong-lanceolate, 4—5 ст. long, 1–1.5 em. broad, tapering below into a short petiole, entire, glabrous, margin and midrib of under surface scabrous; inflorescence axil- lary, cymose, many-flowered, subtended by bracts; calyx deeply mm. longis, apice saepe recurvatis, ciliatis cum duobus generibus multicellularum capillorum, ambis flaccidis subtilibusque; corolla bilabiata, 10-20 mm. longa, rosea vel violacea, lobis obtusis, emarginatis, extus puberulentis, interiore anterioris lobis faucorum pubescente; staminibus parce een antheris pubescentibus; stylo 10-20 mm. longo, stigmata obliqua, lineari; capsula ішпоба.--ТүрЕ collected along the river Apa, Paraguay, Nov. 1901, E. Hassler 7780 (О). 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 31 5-parted, densely hirsute, about 10 mm. long, lobes subulate- setaceous; corolla more ог less bilabiate, approximately 18-20 mm. long, pubescent on the external surface; capsule 8-9 mm. long, 4-seeded, glabrous; seeds flattened, suborbicular. Distribution: southeastern Brazil. Specimens examined: Prov. Goyaz, Brazil, Feb. 1841-42, Gardner 8951 (К түре, М photograph). 4. Dyschoriste angusta (Gray) Small, Fl. Miami, 168. 1913; Small, Fl. Florida Keys, 135. 1913. Calophanes angusta Gray, Syn. Fl. N. Am. ed. 2, 2': 456. 1886; Small, Fl. Southeastern U.S. ed. 1, 1088. 1903, and ed. 2, 1913. Calophanes oblongifolia var. angusta Gray, Syn. Fl. N. Am. ed. 1, 2:: 324. 1878, and ed. 2, 1886. A low-growing perennial, 1-2 dm. high; stem erect or ascending from a creeping base, sligh:;ly puberulent, occasionally branching; leaves many, subsessile, linear to linear-lanceolate, 1.5-2 cm. long, lineolate, entire; bracts foliaceous, about one-half as long as the leaves; flowers axillary; calyx 8-9 mm. long, lobes subulate- setaceous, ciliate, distinct to near the base, hardly surpassing the capsule at maturity; corolla blue, purple, or rarely, white, slightly bilabiate, approxiraately 1 cm. long, tube slightly shorter than the limb and a little ampliated at the base; stamens adnate to the base of the limb о: the corolla; anthers ovate, filaments widening at the base; capsule glabrous, linear, 4-seeded; seeds somewhat oblique. Distribution: southern Florida. Specimens examined: Palm Beach County: Pelm Beach, 26 Dec. 1895-11 Jan. 1896, Hitchcock 1455 (FM). Dade County: pine lands, Grossmanns, 24 May, 1904, Britton 155 (FM); Cocoanut Grove, 26 Dec. 1895, 11 Jan. 1896, Hitch- cock 1456 (FM); Miami, near river, 21 Nov. 1908, Eaton 385 (FM); in pine lands between Cocoanut Grove and Cutler, 13-23 Nov. 1903, Small & Carter 776 (ЕМ); cabbage field in pine woods, Grossmanns, 25 Feb. 1905, Eaton 1247 (С); Biscayne Bay, 1874, Palmer 347 (С, M, US); Miami, May, 1877, Garber (С, M [Vor. 15 32 ANNALS OF THE MISSOURI BOTANICAL GARDEN 120923); Biscayne Bay, June, 1880, Curtiss 1988 (G); Lemon City, 3 March, 1892, Simpson 528 (С, US); rocky, calcareous land, Miami, 6 April, 1897, Curtiss 6868 (M, FM, G, US); Black Point Bridge near Cutler, 27 Feb. 1920, Young 301 (US). 5. Dyschoriste Ригриѕіі! n. sp. PL 5. Perennial, erect or ascending from a suffruticose base, branching quite profusely, more or less pubescent, with short, stout, white hairs; leaves sessile, linear to linear-lanceolate, 18-20 mm. long, 1 mm. or less broad, entire, pubescent; flowers slightly pedicellate, subtended by two foliaceous bracts; calyx 5-parted, 9-11 mm. long, lobes unequal, slightly longer than the tube, pubescent, ciliate; corolla 15-17 mm. long, pubescent on the external sur- face, tube very narrow, not ampliated at the base, approximately 7 mm. long, limb 4 mm. long; anthers of the shorter pair of stamens occasionally smaller; style 11-12 mm. long, stigma ob- lique; capsule linear, glabrous, approximately 10 mm. long; seeds 4, oblique. Distribution: south Mexico. Specimens examined: Puebla: rocky hills, Tehuacan, June, 1905, Purpus 2362 (M TYPE, 05, а, FM); vicinity of San Luis Tultitlanapa, July, 1908, Purpus 3347 in part (M). 6. Dyschoriste lavandulacea (Nees) О. Ktze. Rev. Gen. Pl. 2 486. 1891. Calophanes lavandulaceus Nees in Mart. Fl. Bras. 9: 27. 1847; Nees in DC. Prodr. 11: 112. 1847. Stems erect from а perennial base, 1.5-2 dm. high, sparingly pubescent, quite angular; leaves sessile, linear-lanceolate, 40—50 шигээ Purpusii Kob., вр. поу., caulibus perennis, erectis үе! ascendentibus a suffruticosa basi, ramis profusis, plus minusve pubescentibus cum brevibus albidis capillibus; “foliis шанг, linearibus vel linearo-oblanceolatis, 18-20 mm. longis, mm. minusve latis, integerrimis, pubescentibus; floribus parum pedicellatis, sub- tendentibus bracteis; calyce 5-diviso, 9-11 mm. longo, lobis inaequalibus, paulo tubo longioribus, pubescentibus, ciliatis; corolla extus puberula, 15-17 mm. longa, tubo angustissimo, non ampliato ad basem, plus 7 mm. longo, limbis 4 mm. longis; staminibus postero-lateralibus minoribus; capsula lineari, glabra, 10 mm. longa, 4-врегта.—ТуРЕ collected on rocky hills, Tehuacan, June, 1905, C. A. Purpus 2362 (M). 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 33 mm. long, 4-5 mm. wide, tapering to an acute apex, entire, glabrous; inflorescence somewhat glomerulate, several-flowered; bracts minute, 3-5 mm. long, resembling the calyx in texture; calyx 5-lobed, 13-14 mm. long, tube about 5 mm. long, glabrous and covered with cystoliths except for the ciliate, subulate- setaceous lobes; corolla 5-lobed, slightly bilabiate, 20 mm. long, pubescent on the external surface, tube one-half the total length of the corolla, lobes quite truncate; anther cells slightly puberu- lent; mature capsule not seen. Distribution: south-central Brazil. Specimens examined: In dry fields, near Rio Pardo, Brazil, Sept. 1826, Riedel 501 (L түре, М photograph). 7. Dyschoriste Greenmanii! п. sp. РІ. 6. Plants about 2 dm. high, ascending from a perennial base; stems slender, branched, pubescent at the nodes, otherwise quite glabrous; leaves sessile, linear, 20-25 mm. long, approximately 2 mm. broad, entire, ciliate, sparingly pubescent; flowers few, solitary at the nodes, subtended by 2-foliaceous bracts; calyx deeply 5-parted, 15 mm. long, flaccid-pubescent on the main nerves; lobes subulate-setaceous, approximately 10 mm. long; corolla pubescent on the external surface, 17 mm. long, scarcely exceeding the calyx in length, tube 6.5-7 mm. long, throat more or less equalling the tube in length; style 12 mm. long, pubescent, stigma oblique; capsule linear, glabrous, 7-8 mm. long, 4-seeded. Distribution: northeastern Mexico. Specimens examined: Vicinity of Victoria, Tamaulipas, alt. 320 ш., 1 May-13 June, 1907, Palmer 492 (05 түре, М photograph and fragment). 1 Dyschoriste Greenmanii Kob., sp. nov., planta prope 2 dm. alta, ascendens a perenne basi; caulibus gracilibus, ramosis, pubescentibus ad nodos, aliter glabris; foliis linearibus, sessilibus, 20-25 mm. longis, 2 mm. latis, integerrimis, ciliatis, parce pubescentibus; floribus paucis, solitariis ad nodos, subtendentibus 2-foliaceis brac- teis; calyce profunde 5-diviso, 15 min. longo, flaccido-pubescente nervis, lobis subu- lato-setaceis, prope 10 mm. longis; corolla 17 mm. longa, paulo calyce longiore, tubo 6.5-7 mm. longo, fauce plus minusve aequante tubum; stylo 12 mm. longo, pubes- cente, stigmata obliqua; capsula lineari, glabra, 7-8 mm. longa, 4-sperma.—T yPE collected in the vicinity of Victoris, Tamaulipas, Mexico, 1 May-13 June, 1907, E. Palmer 492 (05). [ Уот. 15 34 ANNALS OF THE MISSOURI BOTANICAL GARDEN 8. Dyschoriste Niederleinii Lindau in Engl. Bot. Jahrb. 19 (Beibl. 48): 15. 1894. Low-growing perennial; stems ascending, about 1 dm. high, branches tetragonal, minutely puberulent; leaves linear, approx- imately 80 mm. long, 5 mm. broad, somewhat obtuse at the apex, entire, glabrous, sparsely pilose at the base, petiole 2 mm. long; flowers single, axillary, subtended by small bracts; calyx 5-parted, puberulent, 11 mm. long, tube and calyx lobes of equal length; corolla puberulent on the external surface, 15 mm. long, ventri- cose; style 6 mm. long, filiform, pubescent; mature capsule unknown. Distribution: Argentina. Specimens examined: “Аа Primer Misionero de Hernandez," Puck and Fernandez (Niederlein 42), Argentina, Feb. 1884, (В түре, М photograph). 9. Dyschoriste Schiedeana (Nees) O. Ktze. Rev. Gen. Pl. 2: 486. 1891. Calophanes Schiedeanus Nees in DC. Prodr. 11: 111. 1847; Mueller in Walpers, Ann. 5: 647. 1858, including var. multi- florus; Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. Perennial, ascending or erect from a suffruticose base, 3-6 dm. high; stems somewhat angular, hirsute, branched near the base; leaves usually linear-lanceolate, lower cauline leaves occasionally obovate, 20-25 mm. long, 3-5 mm. broad, acute at the apex, narrowed at the base into а very short petiole, entire, hirsute on both surfaces; flowers axillary, usually two in an axil, subtended by bracts which equal or nearly equal the calyx in length; calyx 5-parted, 11-12 mm. long, lobes 7 mm. long, subulate-setaceous, hirsute; corolla 14-15 mm. long, pubescent on external surface, tube 4 mm. long; mature capsule 7-8 mm. long, linear, glabrous, acute at apex, 4-seeded; seeds typical. Distribution: eastern Mexico. Specimens examined: Nuevo Leon: near Monterey, alt. 550 m., Aug. 1911, Arséne 6411 (US). Vera Cruz: in fields near Jalapa, date lacking, Schiede 122 (M photograph of type, В түре); Mirador, date lacking, Sartorius (US 55268). 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 35 10. Dyschoriste Rosei! n. sp. Pl. 7, fig. 1. Low-growing perennial; stem pubescent, branched, ascending or erect, 12-15 cm. high; leaves sessile, linear, entire, glabrous, 18—25 mm. long, 2 mm. broad; flowers few, solitary at the nodes, usually near the apex of the stem, slightly pedicellate, subtended by 2-foliaceous bracts; calyx 5-parted, glabrous except for the ciliate margin of the unequal, subulate-setaceous lobes, shorter posterior lobes 8-9 mm. long, anterior lobes 11-12 mm. long; corolla externally pubescent, 25 mm. long, tube 10 mm. long, diverging abruptly into a broadly ampliated throat which is approximately equal the tube in length; stamens occasionally incompletely didynamous: ovary 2-celled, each cell containing 2 ovules, style 17-18 mm. long, stigma oblique, 2 mm. long; mature capsule not seen. Distribution: western Mexico. Specimens examined: Durango: without definite locality, 13 Aug. 1897, Rose 2259 (US түре, М fragment and photograph) Jalisco: on road between Mesquite and Monte Escolebo, 26 Aug. 1897, Rose 3581 (US). 11. Dyschoriste jaliscensis? n. sp. PLS Stems several, 3-4 dm. high, erect from a ligneous, perennial base, branching, pubescen:; leaves linear to linear-oblanceolate, 2.5-3.5 ст. long, 2 mm. or less broad, narrowed at the base, 1 Dyschoriste Козе! Kob., sp. nov., humilis perennis; caule pubescente, ramis ascendentibus vel erectis, 12-15 cm. altis; foliis linearibus, sessilibus, 18-25 mm longis, 2 mm. latis, integerrimis, glabris; floribus paucis, solitariis ad nodos, fere prope apicem, subpedicellatis, subiendentibus bracteis; calyce 5-diviso, glabro, lobis inaequalibus, subulato-setaceis, posterioribus lobis 8-9 mm. longis, EI lobis 11-12 mm. longis; corolla extus puberula, 25 mm. longa, tubo 10 mm. longo, divere gente subito in late ampliato favce; staminibus didynamis, 4 чонй ovario biloculo, stylo 17-18 mm. longo; stigma obliqua, 2 mm. longa; capsula ignota. --ГүрЕ collected in the state of T 13 Aug. 1897, Rose 2259 (US). з Dyschoriste jaliscensis Kob., вр. nov., planta 3-4 dm. alta; caulibus pluribus, erectis а lignoso Pod ramis pubescent foliis Каранов, linearo-oblanceolatis, 2.5-3.5 cm. longis, 2 e latis, basi attenuatis, integerrimis, pubescentibus; floribus maioribus, Заме гај foliaceis, 10 mm. longis; calyce 17-18 mm. longo, pu- bescente, lobis 11-12 mm. longis, оао ciliatis; corolla prope 80 mm. longa, tubo 12-13 mm. longo, pauce fauce longiore; antheris 2 mm. longis, basi bicalcaratis; stylo 20-21 mm. longo, stigmata obliqua, capsula ignota.—T PE col- lected on rocky hills near Guadalajara, Jalisco, 27 June, 1893, Pringle 5481 (С). [Vor. 15 36 ANNALS OF THE MISSOURI BOTANICAL GARDEN entire, pubescent; flowers comparatively large, subtended by 2-foliaceous bracts which are about 10 mm. in length; calyx 17- 18 mm. long, pubescent, lobes 11-12 mm. long, subulate, seta- ceous, ciliate; corolla approximately 30 mm. long, tube 12-13 mm. long, slightly longer than the throat; anthers about 2 mm. long; style 20-21 mm. long, stigma oblique; mature capsule not seen. Distribution: western Mexico. Specimens examined: Jalisco: rocky hills near Guadalajara, 27 June, 1893, Pringle 6481 (US, С түре, М photograph and fragment). 12. Dyschoriste angustifolia (Hemsl.) О. Ktze. Rev. Gen. РІ. 2: 485. 1891. Calophanes angustifolius Hemsl. in Biol. Cent.-Am. Bot. 2: 502. Stem erect, strict, 4-5 dm. tall, more or less villous-hirsute; leaves subsessile, linear-lanceolate, 1.5-2.5 cm. long, 3-5 mm. broad, acute at the apex, attenuate at the base, entire, scabrous; flowers axillary, disposed in dense shortly pedunculate cymes in the axils of the upper leaves, subtended by narrow bracts which almost equal the calyx in length; calyx deeply 5-parted, 15 mm. long, scabrous, lobes subulate-setaceous, nearly equalling the tube of the corolla, ciliate; corolla bilabiate, puberulent on the external surface, approximately 25 mm. long; anther cells shortly mucronate at the base; ovary 2-celled, cells 2-ovulate, glabrous, stigma linear, oblique; mature capsule not seen. Distribution: southern Mexico. Specimens examined: Oaxaca: without definite locality, coll. of 1842, Ghiesbreght (K, M photograph). 13. Dyschoriste linearis (Torr. & Gray) О. Ktze. Rev. Gen. РІ. 2: 486. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 45>; 302. 1895; Lindau in Bull. Herb. Boiss. II. 6: 844. 1906. Dipteracanthus linearis Torr. & Gray, Bost. Jour. Nat. Hist. 5: 50. 1845 (РІ. Lindh. 1: 50). Calophanes linearis Gray, Syn. Fl. N. Am. 2': 324. 1878, and 1928] KOBUSKI—-MONOGRAPH OF DYSCHORISTE 37 ed. 2, 1886; Hemsl. in Bio.. Cent.-Am. Bot. 2: 503. 1882; Small, Fl. Southeastern U. 5. 1083. 1903, and ed. 2, 1913. Calophanes oblongifolius var. texensis Nees in DC. Prodr. 11: 108. 1847; Torr. in Emory’s Rept. U.S. & Mex. Bound. Surv. 2 (Bot.): 122. 1859. Calophanes ovatus Nees in DC. Prodr. 11: 108. 1847, not Ruellia ovata Cav. Ruellia ovata Benth. Fl. Hartweg. 89. 1842, not Cav. i.e., as to plant of Drummond from Texas. Stem 18-42 cm. high, erect and strict, branched and diffuse, hirsute with both rigid and short hairs, sometimes sparsely pubescent or nearly glabrous, not cinereous; leaves linear-oblan- ceolate to oblong-spathulate, 1.8-6.5 cm. long, entire, lineolate, rather rigid, pubescent on midrib and veins, margin ciliate; bracts foliaceous, frequently in short-leafed specimens equalling the length of the leaf; calyx 5-cleft, densely lineolate, giving the appearance of appressed hairs, lobes 9-13 mm. long, subulate- setaceous, more or less hispid, ciliate, calyx tube 4.5-6 mm. long, in most cases one-half the length of the lobes; corolla somewhat bilabiate, 26-27 mm. long, pubescent on external surface, tube 5—7 mm. long and slightly shorter than the abruptly ampliated limb; anther cells oblong; capsule 4-seeded; seeds flat. Distribution: Texas to New Mexico and northern Mexico. Specimens examined: Texas: rocky prairies, 12 July, 1908, Reverchon (M. 120836); western Texas, 1890, Nealley (Ch 254803); 1846, Lindheimer 504 (US); Drummond 2 no. 178 (С түре); Drummond 856 (С); dry prairies, Bay City, Mategorda Co., 6 May, 1916, Palmer 9667 (M); prairies, Ganado, Jackson Co., 20 March, 1916, Curtiss 9216 (M); dry open ground, Vanderbilt, Jackson Co., 10 May, 1916, Palmer 9708 (М); Calhoun Co., 10 Aug. 1920, Drushel 4136 (M); dry rocky prairies near Dallas, Dallas Co., June-July, Curtiss 1941 (FM, M); dry rocky prairies, Dallas, Dallas Co., May-June, 1879, Reverchon (FM. 88468); dry rocky prairies near Dallas, Dallas Co., date lacking, Reverchon 1941 (С, M, US); rocky limestone prairies, Dallas Co., 15 Мау, Reverchon 722 (М, US); Dallas Co., 23 May, 1908, Bebb 1343 (FM); rocky prairies, Dallas Co., 18 May, 1900, Reverchon 2114 (M); field and gardens, Fort Worth, [Vor. 15 88 ANNALS OF THE MISSOURI BOTANICAL GARDEN Tarrant Co., 8 June, 1909, Ruth 104 (US); along roadsides near Fort Worth, Tarrant Co., 5 July, 1909, Ruth 30 (ЕМ); dry grounds near Fort Worth, Tarrant Co., 1 June, 1910, Ruth 81 (FM); Austin, Travis Co., 25 June, 1920, Tharp 733 (US); Austin, Travis Co., 1897, Buckley (М 120803); dry hills, Austin, Travis Co., 13 May, 1872, Hall 431 (G); dry prairies, Austin, Travis Co., 16 May, 1872, Hall 428 (US); along Corpus Christi Bay, Nueces Co., alt. sea level, 9-12 April, 1894, Heller 1529 (G, M US, FM); dry open ground, Strawn, Palo Pinto Co., 27 June, 1918, Palmer 14262a (М); Dublin, Erath Co., 1893, Maxwell 49 (Ch); Round Top Mt., Comanche Co., 9 May, 1900, Eggert (M, 120809); Gillespie Co., date lacking, Jermy 472 (M); rich hillside, Boerne, Kendall Co., 19 May, 1916, Palmer 9811 (M); in dried river beds of mountain rivers north of Braunfels, Comal Co., 1846, Lind- heimer 325 (M); in pastures, Bracken, Comal Co., 3 Aug. 1903, Groth 230 (G); humid prairie and along margin of shrubs near New Braunfels, Comal Co., May, 1848, Lindheimer 677 (G, M, FM, US); in grass and on black prairie loam, New Braunfels, Comal Co., May, 1846, Lindheimer 111 (G, M); Comanche Springs, New Braunfels, Comal Co., May, 1851, Lindheimer 1063 (M, G, FM, US); New Braunfels, Comal Co., May, 1851, Lind- heimer 552 (M); in open pastures, 5 miles south of San Antonio, Bexar Co., 14 May, 1920, Schultz 146 (US); San Antonio, Bexar Co., 1918, Slater (US 891769); San Antonio, Bexar Co., 1884, Havard (Ch 252081); Bexar Co., date lacking, Jermy 62 (M, US); San Antonio, date lacking, Jermy 249 (G); San Antonio, Bexar Co., 27 April, 1911, Mr. & Mrs. Clemens 1069 (M); Bexar Co., date lacking, Jermy 31 (US); San Diego, Duval Co., 1885, Croft 6465 (M); San Diego, Duval Co., July, 1885, Croft 6660 (М); dry open ground, Baird, Callahan Co., 26 May, 1918, Palmer 13698 (M); Abilene, Taylor Co., 22 May, 1902, Tracy 8079 (G, M, FM, US); prairie north of Abilene, Taylor Co., 7 June, 1900, Eggert (M 120800); calcareous banks, Menard Co., 11 May, 1917, Palmer 11871 (M); dry alluvial soil along creek, Lacey’s ranch, Kerr Co., 10 June, 1917, Palmer 12229 (М); rocky ground, Sweet- water, Nolan Co., 27 May, 1918, Palmer 13758 (M); Knicker- bocker ranch, Dove Creek, Tom Green Со., May, 1880, Тиееду 180 (US); Fort Clark, Kinney Co., 10 May, 1893, Mearns 1432 1928] KOBUSKI—MONOGRAPH ОЕ DYSCHORISTE 39 (US); Devils River, Valverde Co., May, 1913, Orcutt 6230 (M); prairie north of Stanton, Martin Co., 13 June, 1900, Eggert (M 120810); western Texas to El Paso, New Mexico, El Paso Co., May-Oct. 1849, Wright 432 (а, FM, US). New Mexico: Slaughter Canyon, Guadalupe Mts., 12-20 Aug. 1924, Standley 40624 (US). Mexico: Sierra Madre, 45 miles south of Saltilla on border of states of Coahuila and Nuevo Leon, July, 1880, Palmer 2033 (G); roadside, Piedras Nigras, Coahuila, May, 1883, Havard (Ch 267840, US 147426); near Huasemote, Durango, 15 Aug. 1897, Rose 3495 (US). 14. Dyschoriste bilabiata (Seemann) О. Ktze. Rev. Gen. РІ. 2: 486. 1891; Lindau in Bull. Herb. Boiss. 7: 575. 1899. РІ. 9. Calophanes bilabiatus Seem. Voy. Н. М. В. Herald, 324. pl. 65. 1852-57; Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. Stems 6-7 dm. high, erect from a perennial base, branching, pubescent; leaves ovate-oblong, 4—5 cm. long, 1.5-2 cm. broad, acute at the apex, narrowed at the base into a petiole, repand- denticulate, densely pubescent on both surfaces; flowers axillary, cymose, cymes pedunculate, 3-5-flowered, subtended by subulate bracts; calyx 5-lobed, 12 mm. long, tube 5 mm. long, pubescent, lobes subulate-setaceous, ciliate; corolla subbilabiate, pale blue, 14 mm. long, tube 5-6 mm. long, subventricose, pubescent on the external surface; filaments hairy; ovary glabrous, style filiform, stigma linear, oblique; mature capsule linear, glabrous, 4-seeded. Distribution: western Mexico. Specimens examined: Cero de Pinal, Sinaloa, Dec. 1848, Seemann. 1618 (K TYPE, М photograph only). 15. Dyschoriste decumbens (Gray) O. Ktze. Rev. Gen. Pl. 2: 486. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 4: 302. 1895. Calophanes decumbens Стау, Syn. Fl. М. Am. ed. 1, 2': 325. 1878, and ed.*2, 1886; Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. (Мог. 15 40 ANNALS OF THE MISSOURI BOTANICAL GARDEN Calophanes oblongifolius 'Torr. Bot. Mex. Bound. Surv. 122. 1855, not Don. Cinereous-puberulent throughout; stems mostly spreading on the ground from a ligneous base, occasionally erect, branched; leaves spatulate to oblanceolate, 2-3 ста. long, 0.5-1.1 ста. broad, entire, apex usually obtuse, sometimes slightly mucronate, base attenuated, often having the appearance of a petiole; flowers few, in foliose, bracteate clusters; calyx 15-20 mm. long, at maturity exceeding the capsule by as much as 10 mm., 5-cleft, tube 5-7 mm. long, lobes subulate-setaceous, hardly twice the length of the tube; corolla purple, 18-20 mm. long, tube a little longer than the throat, slightly ampliated at the base; anther cells oblong, filaments united at the base of the corolla-throat; seeds 4, sub- orbicular and flattened. Distribution: dry soil, western Texas to Arizona, and the plateau region of northern Mexico. Specimens examined: Texas: Chenates region of western Texas, 1889, Nealley 580 (357) (US); infrequent on slopes between Marfa and Alpine, 15 April, 1919, Hanson 638 (US). New Mexico: Valley of the Rio Grande, 1851, Wright 1462, 1463 (М). Arizona: Sonoito Valley, Santa Cruz Со., alt. 1833 m., Aug. 1874, Rothrock 637 (US); Fort Huachuca, 1890, Patzky (US 721394); Fort Huachuca, Cochise Co., Мау, 1892, Wilcox (US 55273, M 120796); roadway, Chiricahua Mts., Cochise Co., alt. 1400 m., 9 Oct. 1907, Blumer 2223 (ЕМ) ; Fort Huachuca, Cochise Co., 1894, Wilcox 150 (US); foothills of Santa Rita Mts., near Greaterville, Pima Co., alt. 1666 m., 16 Sept. 1916, Shreve 4978 (US); plains about Huachuca Mts., Aug. 1882, Lemmon (US 55278); locality lacking, 1875, Rothrock (US 55277); Fort Hua- chuca, Cochise Co., 26 April-21 May, 1890, Palmer 472 (US). Mexico: Sierra Mojado Mts., Coahuila, 19 April, 1892, Jones 374 (US, M); near the border of Coahuila and Nuevo Leon, Feb.—Oct. 1880, Palmer 1009 (US); Saltillo, Coahuila, alt. 1600 m., 1911, Arséne 6472 (US); Saltillo, Coahuila, July, 1880, Palmer 2082 (С); Saltillo, Coahuila, May, 1898, Palmer 125 (US, М); Lerios, 15 leagues east of Saltillo near the border of Coahuila and 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 41 Nuevo Leon, alt. 3000 m., 10-13 July, 1880, Palmer 1010 (15458) (US, M); on road near Colatlan, Zacatecas, 31 Aug. 1897, Rose 8615 (US); exact locality lacking, San Luis Potosi, 1897, Schaffner 854 (647) (US); San Luis Potosi, alt. 2000-2500 m., 1878, Parry & Palmer 699 (FM, M, G, US); near Queretaro, 20-23 Aug. 1909, Rose & Rose 11148 (US); San Andres Mts., Chihuahua, 22 Aug. 1900, Trelease 352 (M); hills near Chihuahua, Chihuahua, 30 Sept. 1886, Palmer 1075 (М); vicinity of Chihuahua, Chihuahua, alt. 1300 m., 1-21 May, 1908, Palmer 208 (US); rocky hills near Chihuahua, Chihuahua, May, 1885, Pringle 66 (US, С, ЕМ); Cosihuiriachie, west of the city of Chihuahua, Chihuahua, 20 Sept. 1846, Wislizenus 185 (M); City of Durango, Durango, 1 Aug. 1898, Nelson 4597 (US); in the vicinity of Durango, Duran- go, April-Nov. 1896, Paliner 309 (FM, US, M, G); Palmer 930 (US, M); Palmer 276 (US, G); Sonora, 8 Sept. 1851, T'hurber 974 (G). 16. Dyschoriste crenulata! n. sp. ГІ, 7, Пр. 2: Stems several, 1-2 dm. high, erect or ascending from a peren- nial, ligneous base, pubescent; leaves more or less spathulate to obovate, 2-3 ст. long, 0.5-1 ст. broad, acute to obtuse at the apex, attenuate at the base, densely cinereous pubescent, margin crenulate; calyx 5-parted, 17-18 mm. long, nearly equalling Ше length of the corolla, tube and lobes of nearly equal length, lobes subulate-setaceous, cinereous, ciliate; corolla 18-19 mm. long, pubescent on the external surface, throat slightly longer than the tube; anthers occasionally unequally didynamous, style 11-12 mm. long, stigma oblique; mature capsule not seen. Distribution: south Texas, south into Tamaulipas. Specimens examined: ! Dyschoriste crenulata Kob., sp. nov., planta 1-2 dm. alta; caulibus pluribus, erectis vel ascendentibus a perenne lignoso basi, pubescentibus; foliis subsessilibus, plus minusve spathulatis vel plerumque obovatis, 2-3 em. longis, 0.6-1 cm. latis, apice acutis vel obtusis, crenulatis, basi attenuatis, cinereo-pubescentibus; calyce 5-diviso, 17-18 mm. longo, prope aequante corollam, tubo lobes aequante, lobis subulato-setaceis, cinereis, ciliatis; corolla 18-19 mm. longa, extus puberula, fauce tubo paulo longiore; staminibus clidynamis, subinde imperfectis; stylo pubescente, 11-12 mm. longo, stigmata obliqua; capsula ignota.—T PE collected on road from "San Fernando to Лшепеу,” state of Tamaulipas, Mexico, 26-27 Feb., 1902, E. W. Nelson 6604 (G). [Vor. 15 42 ANNALS OF THE MISSOURI BOTANICAL GARDEN Texas: Brazos Santiago, 1899, Nealley 124 (357) (US). Mexico: “Зап Fernando to Jimeney," Tamaulipas, 26-27 Feb. 1902, Nelson 6604 (С түре, US isotype, М photograph and fragment). 17. Dyschoriste capitata (Oerst.) O. Ktze. Rev. Gen. Pl. 2: 486. 1891. Calophanes capitatus Oerst. in Vidensk. Meddel. 121. 1854; Mueller in Walpers, Ann. 5: 647. 1858; Hemsl. in Biol. Cent.- Am. Bot. 2: 502. 1882. Stems several, frequently branching and ascending from a ligneous base, 4 dm. high, subtetragonal, often geniculate, pubes- cence becoming more pronounced and flaccid near the apex; leaves obovate, 15-27 mm. long, 8-12 mm. broad, obtuse at the apex, attenuate into a petiole varying from a subsessile condition to 5 mm. in length, entire, ciliate, upper surface hirsute, especially on basal portion of midrib and petiole, sparingly so along midrib and veins of lower surface; flowers congested in heads at the apex of the stem and branches, subtended by oblanceolate bracts, the basal portion invested with long whitish hairs; calyx 9-10 mm. long, 5-lobed, joined for one-third its total length, possessing the same pubescence as the bracts, together giving a distinctly whitish appearance to the inflorescence, lobes subulate-setaceous; corolla 15-16 mm. long, puberulent on the external surface; filaments terminated by an emarginate, muticous prolongation at the apex; staminodium sometimes present; ovary glabrous, stigma linear, oblique; mature capsule glabrous, 8-9 mm. long, 4-seeded. Distribution: онан of southern Mexico. Specimens examine Prov. of San Luis Potosi, 1851, Oersted 808! (C); Sierra de San Felipe, Oaxaca, alt. 2000 m., 15 June, 1897, Pringle 6718 (FM, G, M, US); Valley of Oaxaca, alt. 1550 m., 8 June, 1897, Con- гай ; Gonzales 282 (G). 18. Dyschoriste Pringlei Greenm. in Proc. Ат. Acad. 40: 32. 1905. 1 This citation refers to a OP of the only Oersted specimen of D. capitata found in the Copenhagen Herbarium 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 48 Stems several, 1-2 dm. in length, erect or ascending from a ligneous perennial base, densely hirsute-pubescent or subtomen- tose; leaves lance-elliptie to slightly obovate, 1.5-4 cm. long, 0.5-1.6 em. broad, obtuse ог acute, entire, narrowed below to a subpetiolate base, sparingly hirsute-pubescent on both surfaces; flowers crowded in the axils of the upper leaves, forming а hib capitate, leafy inflorescence; calyx 13-14 mm. long, densely pubescent with white flaccid hirsute hairs, divided to somewhat below the middle, divisions lance-attenuate; corolla tubular- campanulate, 8-4 cm. long, externally pubescent, more or less purplish-maculate, at least in the dried state; stamens adnate to the corolla for about one-half its length, anthers rather conspicu- ously calcarate; ovary glabrous, style pubescent; mature capsule not seen. Distribution: southwestern Mexico. Specimens examined: Barranca of Rio Blanco near Guadalajara, Jalisco, alt. 1500 m., 22 July, 1902, Pringle 11313 (G, FM, US); deep canyons near Guadalajara, Jalisco, 1 July, 1889, Pringle 2907 (G түре, ЕМ, M photograph). 19. Dyschoriste oaxacensis! n. sp. Pl. 10. Stems several, procumbent, ascending from a woody base, 1-2 dm. high, pubescent with lineolations showing through pubes- cence; leaves sessile, oblanceolate, occasionally somewhat spathu- late, 15-20 mm. long, 3-5 mm. broad, obtuse at the apex, ciliate, sparsely pubescent or glabrous, appearing scabrous because of the irregular scattering of cystoliths; flowers axillary, congested at the apex of stem and branches, producing a capitate-like in- florescence, subtended by oblanceolate bracts, approximately 10 1 Dyschoriste oaxacensis Kob., sp. nov., caulibus pluribus, procumbentibus, lineolatis, 1-2 dm. altis; foliis ни libus, ohlancesiatie rare subspathulatis, 15-20 mm. longis, 3—5 mm. latis, ciliatis, pauce pubescentibus; floribus axillaribus, in apice caulis ramorumque capitatim congestis; bracteis plus minusve 10 mm. longis; calyce 2 mm. longo, glabro, cystolithero, lobis subulatis, setaceis, ciliatis, 7 mm. longis; corolla subbilabiata, extus puberula, 20 mm. longa, tubo 7 mm. longo; antheris basi bicalcaratis; stylo lineari, pubescente, 13-14 mm. longo, stigmata lineari, obliqua; capsula 10-11 mm. longa, glabra, 4-sperma; seminibus subrotundatibus, planis, humectatis mucilaginosis.— TYPE collected оп calcareous hills, Las Sedas, Oaxaca, Mexico, 9 July, 1891, Pringle 6712 (С). (Мог. 15 44 ANNALS OF THE MISSOURI BOTANICAL GARDEN mm. long, calyx about 12 mm. long, except for the lobes glabrous and covered with cystoliths, lobes subulate-setaceous, ciliate, 7 mm. long; corolla externally puberulent, 20 mm. long, tube 7 mm. long, somewhat bilabiate; stamens adnate below the middle of the corolla limb; ovary glabrous, style 13-14 mm. long, stigma linear, oblique; mature capsule 10-11 mm. long, glabrous, 4- seeded ; seeds oblique, somewhat rounded, flattened. Distribution: southern Mexico. Specimens examined: Oaxaca: calcareous hills, Las Sedas, alt. 2000 m., 19 July, 1891, Pringle 6712 (М түре, С, ЕМ, US); Las Sedas, alt. 2000 m., 2 June, 1907, L. C. Smith 419 (G); Nochixtlan, alt. 2000 m., 19 June, 1907, Сопгаш 1858 (FM). 20. Dyschoriste pinetorum! п. sp. ELAL Stems erect or ascending from a woody, perennial base, branches often arising from nodes of prostrate or erect growth of previous year, subquadrangular, 20-30 cm. high, nodes frequently 5—6 cm. distant, pubescent especially at the apex; leaves obovate- elliptic, 25-35 mm. long, 10-18 mm. broad, acute to subrotund at the apex, subsessile, attenuate at the base into a very short petiole, entire, ciliate, hirsute on both surfaces, the pubescence confined to midrib and veins on the under surface, veins con- spicuous; flowers disposed in heads at the tips of the stems and branches and subtended by oblanceolate bracts; calyx 11-13 mm. long, divided two-thirds the distance to the base into five subu- late-setaceous, ciliate lobes, pubescence similar to that of the bracts, together giving a canescent appearance to the leafy capitate inflorescence; corolla puberulent on the external surface, 1 Dyschoriste pinetorum Kob., вр. nov., caulibus erectis vel ascendentibus а lignoso perenne basi, subquadrangularis, 20-30 сш. altis, nodis saepe ст. diversis, pubescentibus praesertim apice; ramis saepe бардай ех nodis prostratorum vel erectorum caulorum antecedentis anni; foliis subsessilibus, obovato-ellipticis, 25-35 mm. longis, 10-18 mm. latis, apice acutis vel subrotundis, basi in petiolum brevissimum attenuatis, integris, ciliatis, utrinque hirsutis, praesertim ad costos 11-13 mm. longo, diviso ad 38 a basi in quinque subulatis, setaceis, ciliatis lobis, pubescentibus, canescentibus; corolla extus puberula, 20-21 mm. longa, tubo imbum aequante; stylo hirsuta, stigmata lineari, obliqua; capsula ignota.—T PE collected in sandy fields under pines, near Patzcuaro, Michoacan, 31 July, 1892, C. G. Pringle 4134 (G). 1928] KOBUSKI—MONOGRAPH ОҒ DYSCHORISTE 45 20-21 mm. long, tube and :hroat of approximately equal length; stamens adnate to about the middle of the corolla-tube; style hirsute, stigma linear, oblique; mature capsule not seen. Distribution: southern Мехіко. Specimens examined: Michoacan: sandy fields under pines near Patzcuaro, 31 July, 1892, Pringle 4134 (С түрЕ, isotypes in М, Ch, FM, 05). 21. Dyschoriste sagittata! n. sp. Pl. 12. Low-growing perennial; stems ascending 1-2 dm. high from a ligneous base, glabrous or nearly so, densely covered with cystoliths, quadrangular, somewhat winged, branched; leaves sessile, elliptic-obovate, 15-25 mm. long, 9-12 mm. broad, usually obtuse at the apex and base, glabrous, entire; bracts slender, lanceolate, glabrous, 8 mm. long, bracteoles minute, acuminate, 2-3 mm. long; calyx 8-10 mm. long, minutely pubescent on the nerves, lobes about 5 mm. long, subulate-setaceous, sparsely and minutely ciliate; corolla pubescent on the external surface, barely exceeding the calyx in length, ventricose, slightly bilabiate, lobes rounded, margins crenate; stamens small, filaments adnate to a little below the middle of the corolla-throat, anther cells converg- ing towards the acute apex, slightly diverging at the calcarate base, giving a sagittate appearance, approximately 0.5 mm. long; style 4-5 mm. long, minutely pubescent or glabrous, stigma lobed; mature capsule not seen. Distribution: Paraguay. Specimens examined: Paraguay: in region along the Alta Parana River, 1909-10, Fiebrig 6383 (С түре, М fragment and photograph). 1 Dyschoriste sagittata Kob. sp. nov., humilis perennis; caulibus ascendentibus, 1-2 dm. altis a lignoso basi, glabris, cystolitheris, quadrangularis, alatis, ramosis; foliis sessilibus, elliptico-obovatis, 15-25 mm. longis, 9-12 mm. latis, glabris, apice basique obtusis; bracteis angustis, lanceolatis, glabris, 8 mm. longis, bracteolis minutis, acuminatis, 2-3 mm. lonzis; calyce 8-10 mm. longo, puberulente in nervis loborum calycum, lobis prope 5 mm. longis, subulatis-setaceis, parce et minute ciliatis; corolla extus puberula, minor calyce paululo longiore, ventricosa, subbilabi- ata, lobis rotundis, асық crenatis; antheris sagittatis, basi divergentibus et bicalcaratis, apice acutis; stylo 4-5 mm. longo, parce pubescente vel glabro, stigmata trilobata, medio lobo longissimo, circiter 0.5 mm. longo; capsula ірпоба.--ТүрЕ collected in the region along the Alta Parana River, Paraguay, coll. of 1909-10, Fiebrig 6383 (G) [Vor. 15 46 ANNALS OF THE MISSOURI BOTANICAL GARDEN 22. Dyschoriste Serpyllum (Nees) O. Ktze. Rev. Gen. Pl. 2: 486. 1891. Calophanes Serpyllum Nees in Mart. Fl. Bras. 9: 26. 1847 ; Nees in DC. Prodr. 11: 110. 1847. Stems 1 dm. or less high, erect from a suffruticose base, pubes- cent; lower leaves ovate, upper leaves ovate-lanceolate, narrowed at the base, 10-12 mm. long, 3-5 mm. wide, entire, glabrous, subsessile; flowers few, usually toward the apex of the stem, subtended by leafy, glabrous bracts; ealyx unequally 5-parted, submembranaceous, sparsely pubescent, 12 mm. long, lobes greatly attenuated, approximately twice as long as the tube, ciliated; corolla 12-13 mm. long, pubescent on the external surface, tube very short, not more than 3-4 mm. long, ampliation into throat apparently beginning at the base of the tube ; stamens abruptly yet obtusely appended at the apex, bicalcarate at the base; ovary 2-celled, each cell possessing 2 ovules, style sparsely pubescent, filiform, stigma oblique; mature capsule not seen. Distribution: southeastern Brazil. Specimens examined: Brazil: in dry fields near Rio Pardo, Sept. 1826, Riedel 45 (L TYPE, M fragment and photograph). 23. Dyschoriste Lloydii' n. sp. РІ, 18. Stems branched near the base, erect, pubescent, 1-1.5 dm. high; leaves sessile, oblong-oblanceolate, 18-20 mm. long, 3-4 mm. broad, sparingly hirsute-pubescent on both surfaces, often con- fined to the midrib and veins, entire; bracts foliaceous, nearly equalling the calyx in length; calyx 10-10.5 mm. long, tube 4—5 mm. long, sparingly pubescent, lobes ciliate, subulate-setaceous ^ corolla 14 mm. long, tube 5 mm. long, approximately equalling the throat in length; ovary and stamens typical of the genus; ' Dyschoriste Lloydii Kob. sp. nov., caulibus erectis vel ascendentibus, pubes- centibus, 1-1.5 dm. altis; foliis sessilibus, oblongo-oblanceolatis, 18-20 mm. longis, mm. latis, integris, utrinque parce hirsuto-pubescentibus, praesertim ad costos nervosque; bracteis foliaceis; calyce prope 10-10.5 mm. longo, tubo 4-5 mm. longo, parum pubescente, lobis ciliatis, subulato-setaceis; corolla 14 mm. longa, tubo 5 mm. longo, prope aequante ampliatum faucem; capsula lineari, glabra, 7-8 mm. longa, 4-sperma; seminibus typicibus.—T»PE collected near Hacienda de Cedros, state of Zacatecas, Mexico, 1908, F. E. Lloyd 199 (US). 1928] KOBUSKI—MONOGRAPH ОЕ DYSCHORISTE 47 capsule linear, glabrous, 7-8 mm. long, 4-seeded; seeds flattened, suborbicular, oblique. Distribution: central Mexico. Specimens examined: Zacatecas: flats, Hacienda de Cedros, summer 1908, Lloyd 199 (US түре). 24. Dyschoriste microphylla (Cav.) О. Ktze. in Rev. Gen. РІ. 2: 486. 1891. Pl. 14. Calophanes microphyllus (Cav.) Nees in DC. Prodr. 11: 118. 1847; Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. Ruellia microphylla Cav. Ic. 6: 63, pl. 586, f. 2. 1801; Spreng. Syst. 2: 821. 1825. Dyschoriste Jasminum О. Ktze. in Rev. Gen. Pl. 2: 486. 1891. Calophanes Jasminum-mexicanum Nees in DC. Prodr. 11: 110-111. 1847; Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. Stem rising or ascending from a perennial base, pubescent; leaves distinctly ovate, obtuse-rotund at the apex, attenuate at the base into a petiole, 1.5-3 em. long (including petiole), 0.9-1.2 cm. broad, glabrous except for slight pubescence on midrib and margin, entire; inflorescence terminal or on rather short lateral branches, subtended by foliaceous bracts; calyx 5-parted, 12-13 mm. long, somewhat pubescent, especially on the lobes, lobes subulate-setaceous, one-half the total length of the calyx, ciliate; corolla puberulent on the external surface, 13-14 mm. long, tube 8-9 mm. long, ampliating abruptly into the short throat, lobes rounded; stamens adnate below the center of the corolla throat, filaments broadening toward the base; style filiform, pubescent, stigma ampliated, posterior lobe rudimentary; capsule glabrous, 4-seeded, about 8 mm. long. Distribution: southern Mexico. Specimens examined: Puebla: Chalmo y Sar. Miguel, 1789-1794, D. Luis Née (Ma ТУРЕ, M 928687, photograph); vicinity of Puebla at the Rancho Losado, alt. 2194 m., 29 Aug. 1909, Bro. Nicolas 299 (US); vicinity of Puebla, date and number lacking, Bro. Arséne (US 1004058); Cerro Guadalupe, vicinity of Puebla, alt. 2200 m., June, 1908, Arséne 1933 (M, С, US); entre les haciendas Santa [Vor. 15 48 ANNALS OF THE MISSOURI BOTANICAL GARDEN Barbara et Cristo, sur l'Alseseca, alt. 2150 m., 27 June, 1907, Arséne 1528 (US); Santa Barbara, Puebla, 1 June, 1907, Arséne 1075 (US, M). Mexico: hills in the valley of Mexico, alt. 2500 m., 24 Aug. 1902, Pringle 11322 (G, US). Michoacan: vicinity of Morelia, Punguato, alt. 2100 m., 16 July, 1909, Arséne 3044 (M, US); Arséne 52a (FM, US); Morelia, alt. 2000 m., 4 Aug. 1910, Arséne (US 1134412); vicinity of Morelia, north of Zapote, alt. 1950 m., 4 Aug. 1910, Arséne 5728 (M, G, US); vicinity of Morelia, Cuincho, alt. 1900 m., 1 July, 1909, Arséne 7303 (М, US). 25. Dyschoriste saltuensis Fernald, Proc. Am. Acad. 33: 92. 1898. A slender, erect, suffrutescent plant; stems branching, subtet- ragonal, densely covered with short appressed hairs, ciliate at the nodes; leaves lanceolate, obtuse at the tips, tapering below into a short petiole, the lower cauline, 6 cm. long, 1.5 ста. broad, the upper scarcely half as large, above covered with cystoliths, beneath strigilose-pubescent on the midrib; flowers axillary, solitary or in glomerules of 2 to 5, peduncles 3 or 4 mm. long; bracts minute; calyx hirsute, 8-10 mm. long, divided half way to the base into 5 lance-subulate lobes; corolla light purple, pubescent without, 15 mm. or less in length, the slender tube equalling the calyx and spreading into a campanulate throat; lobes oblong, truncate, 4 mm. long; filaments hirsute, style hirsute; mature capsule approximately 10 mm. long, glabrous : seeds 4, flat, oblique. Distribution: mountains of southwestern Mexico. Specimens examined: Guerrero: vicinity of Acapulco, Oct. 1894—March 1895, Palmer 606 (С түре, М, FM, Ch, US). 26. Dyschoriste ciliata (Nees) O. Ktze. Rev. Gen. Pl. 2: 486. 1891; Lindau in Engl. Bot. Jahrb. 19 (Beibl. 48): 15. 1894. Calophanes ciliatus Nees in DC. Prodr. 11: 110. 1847. Ruellia ciliata Ruiz in DC. Prodr. 11: 110. 1847. Stem procumbent, glabrescent, with the apex and ascending 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 49 branches puberulent; lower leaves more or less spathulate, 3 ст. long, 1 cm. broad, upper leaves ovate to elliptic, 6-7 cm. long, 2-3 cm. broad, obtuse to acute at the apex, entire, nearly glabrous, the base cuneate-decurrent into a petiole about 1.5 cm. long; flowers axillary, in glomerules, subsessile, subtended by oblong, ciliate bracts; calyx 5-parted, 11 mm. long, joined for more than one-half the total calyx-length, lobes subulate-setaceous, ciliate; corolla infundibuliform, a little longer than the calyx; anthers slightly bicalcarate at the base. Distribution: Peru. Specimens examined: Peru: near Huanuco, 1757, Ruiz (В түре, M fragment and photograph, 927773). 27. Dyschoriste quadrangvlaris (Oerst.) О. Ktze. Rev. Gen. РІ. 2: 486. 1891; Greenm. in Proc. Am. Acad. 33: 487. 1898. Calophanes quadrangularis Oerst. Vidensk. Meddel. 120. 1854; Mueller in Walpers, Ann. 5: 647. 1858; Hemsl. in Biol. Cent.- Am. Bot. 2: 503. 1882. Stem erect, 8-10 dm. high, distinctly quadrangular, with cili- ated wings; cystoliths especially at the swollen nodes which are quite distant; leaves ovate, oblong, 7-10 em. long, 2-3 em. broad, acute at the apex, attenuate at the base into a petiole, repand, crenulate; flowers verticillate, in cymose clusters at the nodes; calyx 5-parted, 11 mm. long, subtended by short, subulate bracts, tube glabrous, lineolate, equalling or a little shorter than the lobes which are subulate-setaceous, canescent-pubescent along the nerves, ciliate; corolla subbilabiate, 11 mm. long, tube slightly shorter than the limb; stamens adnate to the middle of the corolla- tube; anthers oblong with basal appendages about 0.5 mm. long, filaments accrescent at point of attachment; capsule lanceolate, 8 mm. long, 4-seeded. Distribution: eastern Mexico. Specimens examined: San Luis Potosi: Los Canoas, 29 Aug. 1891, Pringle 5020 (G). Vera Cruz: Potrero de Consoquitla, Nov. 1841, Liebmann (G СОТУРЕ); rocky soil, Zacuapan and vicinity, Oct. 1906, Purpus 2263 (M, G, FM). [Vor. 15 50 ANNALS OF THE MISSOURI BOTANICAL GARDEN 28. Dyschoriste quitensis (HBK.) O. Ktze. Rev. Gen. Pl. 2: 486. 1891; Lindau in Bull. Herb. Boiss. 5: 679. 1897. Calophanes quitensis (HBK.) Nees in DC. Prodr. 11:110. 1847. Ruellia quitensis HBK. Nov. Gen. 2: 240. 1817; Kunth, Syn. 2:37. 1837. Stem procumbent or ascending from a woody base, 8-4 dm. high, branched, somewhat quadrangular, puberulent; leaves oblong, elliptic-lanceolate, narrowed acutely at both ends, 25- 32 mm. long, 10-12 mm. broad, entire, puberulent, constricted below into a short petiole; flowers axillary, subtended by lance- olate bracts which about equal the calyx in length; calyx 5- parted, 7-8 mm. long, united for about one-half the total calyx- length, nearly as long as the corolla, lobes subulate-setaceous, quite hirsute; corolla 8-9 mm. long, pubescent on the external surface; stamens and pistils typical of the genus; mature capsule 7-8 mm. long, acute at the apex, 4-seeded; seeds typical. Distribution: mountains of Andes, Ecuador. Specimens examined: Ecuador: near Quito, around Panecilli, alt. 3000 m., Humboldt (B туре, M photograph); in Andes of Ecuador, 1857-59, Spruce 5989 (G). 29. Dyschoriste maranhonis (Nees) O. Ktze. Rev. Gen. PE 2: 486. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 499; 302. 1895. Zahlbrucknera maranhonis Pohl. in Mart. Fl. Bras. 9:26. 1847; DC. Prodr. 11: 108. 1847. Calophanes maranhonis Nees in Mart. Fl. Bras. 9: 26. 1847; Nees in DC. Prodr. 11: 108. 1847. Ruellia viscosa Pavon in Mart. Fl. Bras. 9: 26. 1847; DC. Prodr. 11: 108. 1847. Stem ascending or erect from a perennial base, 1-1.5 dm. high, pubescent, densely so at the apex, branched; leaves oblong- oblanceolate, 20-25 mm. long, 6-8 mm. broad, obtuse at the apex, tapering to a short-petiolate base, crenulate-subrepand, lower leaves hirsute, densely so on veins of lower surface, upper leaves subtomentose; flowers axillary, subtended by lanceolate bracts; calyx usually 5-parted, 7-8 mm. long, united for about one-half 1928] KOBUSKI—MONCGRAPH OF DYSCHORISTE 51 the total calyx-length, pubescent, lobes subulate-setaceous, ciliate with long flaccid hairs; corolla approximately 14 mm. long, tube gradually ampliated into the throat, pubescent on the internal as well as the external surface, occasionally only 4-lobed; stamens occasionally incompletely didynamous, the anthers sagittate, appendaged at both ends, acutely appendaged at the base; ovary glabrous, style filiform, pubescent, stigma coiled; capsule linear, 9-10 mm. long; seeds 4. Distribution: southeastern Brazil. Specimens examined: Brazil: St. Ignacio, date lacking, Sellow (В түре, М fragment and photograph). 30. Dyschoriste hirsuta (Oerst.) О. Ktze. Rev. Gen. РІ. 2: 486. 1891. Calophanes hirsutus Oerst. in Kjoeb. Vidensk. Meddel. 71. 1877-78. Robust perennial, вшћти сове at the base; stems erect, 4-6 dm. high, branched, at firs; pubescent, glabrate; leaves ovate, 15-20 mm. long, 9-12 mm. broad, subrepand, pubescent on both surfaces, petiolate, with the petiole 2-3 mm. long; flowers axillary, subtended by two oblanceolate bracts; calyx 5-parted, approxi- mately 10 mm. long, lobes united for one-third the total calyx- length, subulate-setaceous, pubescent on the nerves, ciliate; corolla 15-16 mm. long, pale violet, pubescent on the external surface, tube and throat approximately equal in length; stamens and ovary typical of the genus. Distribution: southeastern Brazil. Specimens examined: Brazil: on fields between Serra da Piedade and Lagoa Santa, 2 May, 1864, Warming (С түре, M fragment and photograph). 31. Dyschoriste hygrophiloides (Nees) О. Ktze. Rev. Gen. РІ, 2: 486. 1891; Lindau in Ето. & Prantl, Nat. Pflanzenfam. 435; 302. 1895. Calophanes hygrophiloides Nees in Mart. Fl. Bras. 9: 26. 1847; Nees in DC. Prodr. 11: 109. 1847. Stems geniculate, ascending from a ligneous perennial base, [Vor. 15 52 ANNALS OF THE MISSOURI BOTANICAL GARDEN 3-4 dm. high, pubescent; leaves petiolate, 2-3.5 cm. long, 1-2 cm. wide, lower leaves obovate-subrotund, more or less emarginate at the apex, attenuate at the base into a rather short petiole, upper leaves elliptic-ovate, softly pubescent on both surfaces, margin somewhat sinuous; inflorescence axillary, in glomerules of 2-5 flowers; bracts linear-oblanceolate, setaceous, pubescent, shorter than the calyx, resembling calyx-lobes, bracteoles present; calyx 5-parted, total length approximately 13 mm., lobes 7-8 mm. long, extremely setaceous, villous-ciliate; corolla somewhat bilabiate, puberulent on the external surface, 13-14 mm. long, tube and throat of about equal length; stamens adnate below the middle of the corolla limb, anther cells ovate; style filiform, quite pubescent, 4 mm. or less in length; stigma curved, mature capsule not seen. Distribution: southeastern Brazil. Specimens examined: Brazil: in grassy fields, Parana, 10 Oct. 1914, Dusén 15640 (G, M photograph). 32. Dyschoriste repens (Nees) О. Ktze. Rev. Gen. РІ. 2: 486. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 439: 302. 1895. Calophanes repens Nees in DC. Ргодг. 11: 109. 1847. Ruellia repens Ruiz ace. to Nees in DC. Prodr. 11: 109. 1847, in synonymy. Stem spreading, ascending, geniculate from a perennial base, densely pubescent, branching; branches rather short, ascending, densely foliate; leaves obovate (lower) to ovate (upper), 2.5-3.5 em. long, approximately 1 em. broad, obtuse to acute at the apex, tapering at the base to a distinct petiole, ciliate, entire, hirsute on the upper surface, pubescence of unequal hairs on the midrib of the under surface; flowers axillary, subtended by foliaceous bracts; calyx 5-parted, 10-11 mm. long, nearly equalling the corolla, lobes united for about one-half the total length of the calyx, subulate-setaceous, conspicuously ciliate; corolla 11-12 mm. long, pubescent on the external surface, tube comparatively broad, short; ovary 2-celled, style filiform, pubescent, 6 mm. long, stigma oblique; capsule not seen. Distribution: Peru. Specimens examined: 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 53 Peru: “near Cheuchin," cate lacking, Ruiz (B түре, М frag- ment and photograph). 33. Dyschoriste Pulegium /Nees) О. Ktze. Rev. Gen. РІ. 2: 486. 1891. Calophanes Pulegium Nees in Mart. Fl. Bras. 9: 25. 1847; Nees in DC. Prodr. 11: 109. 1847. Stem erect from a suffruticose base, subvelutinous-pubescent ; leaves sessile, obovate, 2-2.5 ст. long, approximately 1 cm. broad, obtuse at the apex, tapering at the base, crenulate; flowers axillary in sessile glomerules; calyx 5-parted, 9-10 mm. long, hirsute, joined one-quarter the distance from the base, lobes subulate-setaceous, ciliate; corolla about twice the total calyx length; stamens and pistils typical of the genus; mature capsule lanceolate, 4-seeded. Distribution: southeasterr. Brazil. Specimens examined: Brazil: date and exact locality lacking, Sellow 173 (B TYPE, M photograph). 34. Dyschoriste ovata (Cav.) О. Ktze. Rev. Gen. Pl. 2: 486. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 435: 302. 1895; Lindau in Bull. Herb. Boiss. 5: 678. 1897. Calophanes ovatus Benth. in DC. Prodr. 11: 108. 1847, as to Cavanilles plant (not Hartweg plant); Hemsl. in Biol. Cent.-Am. Bot. 2: 502. 1882. Ruellia ovata Cav. Ic. 3: 28, pl. 254. 1794; Willd. Sp. Pl. 3: 363. 1801. Erect perennial, 4-6 dm. high; stems quite quadrangular, pubescent, sometimes winged, occasionally branched; leaves ovate, obovate or occasionally elliptical, 3-4 cm. long, 1-1.7 cm. broad, obtuse at the apex, attenuated at the base into a very short petiole, quite glabrous with the exception of occasional stiff hairs on the midrib and veins, cystoliths abundant, giving the appearance of appressed hairs, margin entire and ciliated, some- times slightly repand-crenulate; flowers crowded among the foliaceous bracts at the nodes, giving a glomerulate appearance; calyx 5-parted, quite glabrous, covered with cystoliths, 14-15 (VoL, 15 54 ANNALS OF THE MISSOURI BOTANICAL GARDEN mm. long, joined for about one-third the total calyx-length, lobes subulate-setaceous, ciliate; corolla 20-25 mm. long, tube less than 10 mm. long, puberulent on the external surface; stamens, pistil, and fruit typical of the genus. Distribution: southern Mexico. Specimens examined: Vera Cruz: Nogales, Mt. Orizaba, alt. 1400 m., 16 Aug. 1891, Seaton 392 (Ch, G); Borrego, near Mt. Orizaba, 26 Aug. 1866, Bourgeau 2903 (US, G). Morelos: near Cuernavaca, 30 July, 1906, Pringle 13838 (US); near Cuernavaca, alt. 1500 m., 28 July, 1896, Pringle 7249 (US). Michoacan: Morelia, alt. 2100 m., 8 Aug. 1912, Arséne 9027 (US). 35. Dyschoriste amoena (Nees) O. Ktze. Rev. Gen. Pl. 2: 485. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 47: 302. 1895. Calophanes amoenus Nees in Mart. Fl. Bras. 9: 27. 1847; Nees in DC. Prodr. 11: 110. 1847. Stem ascending from a perennial base, approximately 3 dm. high, geniculate, branched, puberulent toward the apex, other- wise glabrous; leaves narrowly oblong-ovate to slightly oblong- oblanceolate, 4-5 em. long, 1-1.5 em. broad, obtuse to acute at the apex, entire, glabrous, attenuate at the base into а very short petiole; inflorescence axillary, bracteate, glomerulate toward the apex; calyx deeply 5-parted, 2 cm. long, quite robust, pubescent, lobes less attenuate than in the majority of species, ciliate; corolla 5 mm. or more longer than the calyx, pubescent on the external surface; stamens &nd ovary typical of the genus; mature capsule not seen. Distribution: southeastern Brazil. Specimens examined: Brazil: date and exact locality lacking, Sellow (В түре, M photograph and fragment). 36. Dyschoriste xylopoda' n. sp. РІ. 15. ' Dyschoriste xylopoda Kob., sp. nov., caulibus erectis vel ascendentibus, basi crasse lignose, 2-3 . altis, villoso-pabessentibus; folis sessilibus, lanceolato- oblongis, inferioribus тате ovatis, 2.5-3.5 cm. longis, 1 cm. minusve latis, integerrimis; 1928] KOBUSKI-—MONOGRAPH OF DYSCHORISTE 55 Stems strict, rising from а thick woody base to а height of 2-3 dm., villous-pubescent throughout; leaves sessile or nearly so, lanceolate-oblong, or the lowermost occasionally ovate, 2.5-8.5 ста. long, approximately 1 cm. or less broad, entire; flowers 2-3 on а short peduncle in the axils of the leaves, subtended by linear- lanceolate bracts which are a little shorter than the calyx; calyx deeply 5-cleft, 17-18 mm. long, lobes 11-12 mm. long, subulate- setaceous, villous-ciliate; corolla 25-27 mm. long, pubescent on the external surface, tube 10 mm. long; stamens adnate below the middle of the corolla limb, anthers oblong-ovate, 2-3 mm. long; ovary 2-celled, glabrous, style filiform, 20 mm. long, pubes- cent, stigma linear, oblique, nearly 2 ram. long; mature fruit not seen. Distribution: southern Mexico. Specimens examined: Jalisco: hills near Guadalajara, 19 July, 1893, Pringle 4442 (M түре, С, ЕМ). 37. Dyschoriste oblongifolia (Michx.) O. Ktze. Rev. Gen. Pl. 2: 486. 1891; Lindau in Engl. & Prantl, Nat. Pflanzenfam. 43°; 302. 1895. 1,9, Ruellia oblongifolia Michx. Fl. Bor.-Am. 2: 23. 1805; Pursh, Fl. Am. Sept. 2: 420. 1814. ? Ruellia biflora L. Sp. Pl. 2: 635. 1753 (a doubtful synonym —refer to D. Don in Sweet’s Brit. Fl. Garden). Calophanes oblongifolia (Michx.) D. Don in Sweet, Brit. Fl. Gard. 2: pl. 181. 1833; Gray, Syn. Fl. М. Am. ed 1, 2': 324. 1878, and ed. 2, 1886; Chapman, Fl. Southeastern 0.5. ed. 3, 365. 1897; Britt. & Brown, Ill. Fl. 3: 202. 1898; Small, FI. Southeastern U.S. ed. 1, 1083. 1908, and ed. 2, 1913. Dipteracanthus biflorus Nees in Linnaea 16: 294. 1842. Dipteracanthus oblongifolius Chapman, Fl. Southeastern U.S. ed. 2, 303. 1889. floribus 2-3, pedicellatis, axillaribus; braeteis linearo-lanceolatis, calyce haud paulo breviore; calyce profunde 5-diviso, 17-18 mm. longo, lobis 11-12 mm. longis; subu- lato-setaceis, villoso-ciliatis; corolla 25-27 mm. longa, extus puberulenta, tubo 10 mm. longo; filamentis basi connatis, antheris ovato-oblongis, 2-3 mm. longis; stylo 20 mm. longo, pubescente, stigmata lineari, obliqua, prope 2 em. longa; capsula ignota. --Түре collected on hills, near Guadalajara, Jalisco, Mexico, 19 July 1893, С. С. Pringle 4442 (M). (VoL. 15 56 ANNALS OF THE MISSOURI BOTANICAL GARDEN Herbaceous perennial; stem quadrangular, branched at the base, erect, pubescent or softly hirsute, 4-8 dm. high; leaves sessile or short-petiolate, oblong-ovate, 2.5-4.5 em. long, 0.5-1.5 em. broad, rounded or obtuse at the apex, narrowed at the base, entire or slightly crenulate, softly hirsute; flowers solitary, axillary on very short pedicels, subtended by narrowly oblong, leafy bracts; calyx 15-18 mm. long, deeply 5-parted, subulate-setace- ous, lobes ciliate; corolla blue, usually purple-maculate in the throat, approximately 25-27 mm. long, the tube shorter than the abruptly ampliated throat, puberulent on the external surface, lobes rounded; filaments slightly pubescent at point of adnation; anther cells oblong; mature capsule 10-12 mm. long, lanceolate; seeds 4, flattened, oblique. Distribution: sandy pine barrens, southern Virginia to Florida. Specimens examined: Virginia: date and locality lacking, probably southeastern portion of the state, Thurber (G). South Carolina: sandy ground, north of Graniteville, Aiken Co., 21 Мау, 1899, Eggert (M); locality lacking, Мау, 1867, Ravenel (G, US); barrens near Beaufort, Beaufort Co., 26 April, 1917, Churchill 743 (M). Georgia: sand hills between Grovetown and Forrest, Columbia Co., 10 June, 1902, Harper 1312 (M, G, US); low places north of Belair, Richmond Co., 22 May, 1899, Eggert (M); Ocmulgee River swamp below Macon, Laurens Co., Small (FM); Savannah, Chatham Co., 1842, Curtis (M); Darien Junction, McIntosh Co., 31 Мау, 1909, H. H. Smith 2219 (FM); Darien Junction, McIn- tosh Co., alt. sea level, 25-27 June, 1895, Small (FM); Lexington, Oglethorpe Co., 1836, Short (M). Florida: sandy pine lands, date and exact locality lacking, Mohr (US 721388); eastern Florida, 1895, Curtiss (US); pine barrens, Duval Co., 21 April, 1902, Fredholm 5101 (G, US); pine barrens, Duval Co., 28 April, 1902, Fredholm 5127 (G); near Jacksonville, Duval Co., 2 May, 1893, Curtiss 4428 (M, US); dry pine barrens, near Jacksonville, Duval Co., 8 May, 1894, Curtiss 4667 (ЕМ, US); Jacksonville, Duval Co., April, 1869, Canby (M, G, US); dry pine barrens, near Jacksonville, Duval Co., Curtiss 1938 (М, С, US); south Jacksonville, Duval Co., 7 April, 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 57 1897, Churchill (G); dry sandy pine barrens, St. Augustine, St. Johns Co., May-Aug. 1875, Reynolds (M); sandy pine barrens, DeLand, Volusia Co., date lacking, Harkness (M); dry pine woods, DeLand, Volusia Co., 7 May, 1910, Hood (M); vicinity of Eustis, Lake Co., June-July, 1894, Hitchcock 1454 (FM, M); Eustis, Lake Co., 26 April, 1896, Webber 520 (M); sandy soil, high pine lands, vicinity of Eustis, Lake Co., Nash 184 (M, G, 05); Eustis, Lake Co., 28 May-15 June, 1895, Nash 1774 (US); Winter Park, Orange Co., March, 1900, Huger 19 (M); Clarcona, Orange Co., 18-22 Aug. 1899, Pieters 121 (US); dry sand, Okee- chobee region, Brevard Со., 2 June, 1903, Fredholm 5870 (G); Lake City, Jefferson Co., June-July, 1898, Hitchcock 1452, 1453 (FM); Rosewood, Levy Со., June, 1876, Garber (FM); dry sandy ground, Polk Co., 12 April, 1894, Ohlinger 1415 (FM); dry land, Polk Co., 11 June, 1894, Ohlinger 188, 1437 (FM); Lake Alfred, Polk Co., 11 June, 1922, Armstrong & Armstrong (M); Polk Co., March, 1890, Milligan (U3); pine barrens, Tampa, Hillsborough Co., Aug. 1898, Ferguson М); in pine lands near St. Petersburg, Pinellas Co., 10 Nov. 1907, Deam 2832 (а); pine woods, Manatee Co., 16 March, 1887, Rothrock (FM 160054, 322461); sandy field, Bradentown, Manatee Co., 15 May, 1900, Tracy 6683 (M); Fort Myers, Lee Co., 1904, Westgate 3607 (FM); Fort Myers, Lee Co., July-Aug. 1900, Hitchcock: (FM); Aspalaga, Liberty Co., May, 1898, Chapman (M). Alabama: date and locality lacking, Buckley (M, US). 37a. Dyschoriste oblongifolia (Michx.) О. Ktze. forma glabra n. f. Stem and leaves glabrous; otherwise as the species. Distribution: Florida. Specimens examined: Florida: Tocoi, St. Johns Co., 1874, Palmer 346 (M, С); Lake City, Columbia Co., 4 May, 1893, Rolf 190 (M, FM); Gaines- ville, Alachua Co., 5 June, 1910, Hood (M); Fort Myers, Lee Co., July-Aug. 1900, Hitchcock (M); сургезз swamp and low pine land, vicinity of Fort Myers, Lee Co., 8 May, 1916, J. P. Standley 179 (M, G, FM, US); in pine land, Mullock Creek District, about 8 miles southeast of Fort Myers, Lee Co., May-June, 1917, J. P. Standley 443 (M, С, FM, US); in pine land, vicinity of Fort (Мог. 15 58 ANNALS OF THE MISSOURI BOTANICAL GARDEN Myers, Lee Co., 12 May, 1916, J. P. Standley 13 (M, G, FM, US); pine woods, vicinity of Fort Myers, Lee Co., 28 Feb. 1916, J. P. Standley 12852 (US); vicinity of Fort Myers, Lee Co., 29 Feb. 1916, J. P. Standley 12917 (US); moist pine lands, vicinity of Fort Myers, Lee Со., 14 Dec. 1919, P. С. Standley 18894 (US); high pine land, Jessamine, 17-20 April, 1899, Barnhart 2680 (FM). 38. Dyschoriste humilis (Griseb.) Lindau in Engl. Bot. Jahrb. 19(Beibl. 48): 15. 1894; Lindau in Bull. Herb. Boiss. П. 3: 628. 1903. Ruellia geminiflora Kth. var. humilis Griseb. in Pl. Lor. 176. 1874, and Symb. Argent. 259. 1879. Stem slender, branched below, ascending from a ligneous, perennial base, pubescent; leaves oblong-elliptic, 2-9.5 спа. long, 0.4-0.5 ста. broad, tapering acutely both at the apex and at the short-petiolate base, puberulent, ciliate, entire or sinuous; flowers in twos or threes, axillary, subtended by foliaceous bracts about 8-10 mm. long; calyx in anthesis approximately 14 mm. long, puberulent, lobes setaceous, 8 mm. long, at maturity the calyx sometimes attains a length of 20 mm.; corolla 21-22 mm. long, puberulent on the external surface, tube shorter than the broadly ampliated (8-9 mm. in diameter) throat; stamens adnate below the middle of the corolla throat, anther cells oblong, a little over 2 mm. long, cells of individual anthers often differing, one base distinctly acute and minutely apiculated, the other base blunt or slightly mucronate, apex of cells slightly acute; style sparsely pubescent, filiform, 19 mm. long, stigma about 2 mm. long, posterior lobe rudimentary; mature capsule exceeding the calyx lobes, 10 mm. long; retinaculum in center of each cell; seeds two, flat, oblique. Distribution: Argentina. Specimens examined: Argentina: Chaco Santafichna, Mocovi, 5 Хоу. 1903, Venturi 55 (US); Cordoba, Dec. 1891, Kuntze (US 701502); Cordoba, 21 Dec. 1876, Hieronymus (FM 51116, US 282198); near the city of Cordoba, 1874-75, Hieronymus (FM 51115а, US 282197). 1928 KOBUSKI—MONOGRAPH OF DYSCHORISTE 59 39. Dyschoriste paraguariensis! n. sp. Pl. 16. Stems erect, 2-3 dm. high, strict, somewhat quadrangular, nearly glabrous, sparsely pubescent at the nodes; leaves sessile, lanceolate-elliptic, 2-2.5 cm. long, 0.5-0.8 cm. wide, acute at the apex, glabrous, covered with an irregular scattering of cystoliths, margins entire, not ciliated; flowers in twos, axillary, subtended by two foliaceous bracts) which resemble the leaves in nearly every respect; calyx glabrous, except for the ciliation on the lobes, covered with a regular array of cystoliths, 14 mm. long at maturity, lobes lanceolate, setaceous, 10 mm. long; corolla approximately 18 mm. dng, puberulent on the external surface, tube about 9 mm. long; style filiform, about 12 mm. long, pubes- cent, stigma oblique; mature capsule linear, 11-12 mm. long, glabrous, 4-seeded; seeds flattened. Distribution: Paraguay. Specimens examined: Paraguay: in region of the river “Тарігариау,” Aug., Hassler 4855 (С, TYPE). 40. Dyschoriste Tweediana (Nees) О. Ktze. Rev. Gen. РІ. 2: 486. 1891. Calophanes Tweedianus Nees in DC. Prodr. 11: 108. 1847. Stem ascending from a perennial base, 4-6 dm. high, pubescent; leaves ovate-elliptie, 3-3.5 cm. long, 0.9-1.5 ст. broad, acute to obtuse at the apex, tapering at the base into a very short petiole, repand-subcrenate, glabrous; flowers axillary, 1-3 aggregated on very short peduncles in the axils, subtended by oblong-lanceolate bracts which are shorter than the calyx; calyx deeply 5-parted, pubescent, 14 mm. long, lobes subulate-setaceous, ciliate; corolla infundibuliform, 5-lobed, 20 mm. long, tube 8 mm. long, pu- bescent on the external surface, lobes ovate, obtuse; anthers 1 Dyschoriste paraguariensis Кођ. sp. nov., caulibus erectis, 2-3 dm. altis, strictis plus minusve quadrangularis, subglabris, parum pubescentibus ad nodos; foliis sessilibus, lanceolato-ellipticis, ‘22.5 cm. longis, 0.5-0.8 cm. latis, apice acutis, glabris, lineolatis; floribus duobus in axillaribus, bracteis duobus, foliaceis; calyce glabro, cystolithero, 14 mm. lonyo, lobis lanceolatis, setaceis, 10 mm. longis; corolla violacea, 18 mm. longa, extus puberula, tubo 9 mm. longo; filamentis basi connatis; stylo 12 mm. longo, piloso; capsula lineari, 11-12 mm. longa, glabra, 4-sperma; seminibus planis.— T Px collected in the region of the river **Tapiraguay," Paraguay, Aug., Hassler 4355 (G) “ог. 15 60 ANNALS OF THE MISSOURI BOTANICAL GARDEN appendaged at the base, appendages connivent (according to Nees); seeds suborbicular, flattened. Distribution: southeastern Brazil. Specimens examined: Brazil: in dry mountain forests in Prov. Bonar, at river Jacuhy in Rio Grande do Sul, date lacking, Tweedie 771 (К түре, М photograph). EXCLUDED SPECIES Calophanes californica Rose acc. to Vasey & Rose in Contr. U.S. Nat. Herb. 1:85. 1890 = Ruellia californica (Rose) Johnston in Proc. Cal. Acad. Sei. IV, 12: 1171. 1924. Calophanes cubensis A. Rich. in Sagra, Hist. de Cuba 11: 160. 1850 = Hygrophila brasiliensis (Spreng.) Lindau in Urb. Symb. Ant. 2: 183. 1900. Calophanes Palmeri Gray асс. to Watson in Proc. Am. Acad. 22: 443. 1887 = Spigelia scabrella Benth. Pl. Hartweg. 45. 1840. Calophanes peninsularis Rose ace. to Vasey & Rose in Contr. U. 5. Nat. Herb. 1: 75. 1890 = Ruellia peninsularis (Rose) Johnston in Proc. Cal. Acad. Sci. ТУ, 12: 1172. 1924. Dyschoriste candida Brandegee in Zoe 5: 242. 1908 = Ruellia candida (Brandegee) Kobuski, n. comb. Dyschoriste cubensis Urb. Symb. Ant. 7: 381. 1912 = Apas- salus cubensis (Urb.) Kobuski in Ann. Mo. Bot. Gard. 15:2. 1928. Dyschoriste diffusa Urb. Symb. Ant. 7: 380. 1912 = Apas- salus diffusus (Urb.) Kobuski in Ann. Mo. Bot. Gard. 15:1. 1928. Dyschoriste humistrata (Shuttl.) О. Ktze. Rev. Gen. РІ. 2: 486. 1891 = Apassalus humistratus (Shuttl.) Kobuski in Ann. Mo. Bot. Gard. 15: 3. 1928. List or EXSICCATAE The distribution numbers are printed in italics. Unnumbered collections are indicated by a dash. The number in parentheses is the species number used in this monograph. Armstrong, Dr. & Mrs. G. „~ -- am Blumer, J. C. 2223 (15). Arséne, Bro. 6411 (9); 6472 (15); Bourgeau, M. 1262 (1); 2903 (34). $5, 52a, 1075, 1528, 1933, на 5728, Britton, М. L. 155 (4). 7808 (24), 9027 (34). Buckley, 5. В. — (13); — (37). Barnhart, 7. Н. 2680 (37а). Сапђу, У. М. — (37). Веђђ, В. 1348 (13). Сћартап, С. У. — (37). 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 61 Churchill, J. R. —, 743 (37). Clemens, Mr. & Mrs. J. 1069 (13). Conzatti, C. 1521 (1); 1858 (19). Conzatti, C. & Gonzalez, V. 282 (17). Croft, Miss M. B. 6465, 6660 (13). Curtis, M. — (37). Curtiss, A. H. 5858 (4); 1941, 9816 (13); —, 1988, 4428, 4667 (37). Den, Mrs. C. C. 2832 (37). Drummond, T. 178, 256 (13). Drushel, J. A. 4136 (13). Dugés, A. — А Dusén, Р. 15640 (31). Eaton, А. А. 385, 1247 (4). Eggert, H. —, — (13); — (37). Ehrenberg, К. 1223 (1). Ferguson, А. М — (37). Fiebrig, К. 4856 2); 6383 (21). Fredholm, A. 6101, 5127, 5870 (37). Gonzalez, V. 43 (1). Groth, H. A. 230 (13). Hall, E. 428, 431 (13). Hanson, H. C. 638 (15). (37). Hassler, E. 5908, 7780 (2); 4356 (39). Малын У. — 13). Heller, A. A. 1529 (13). Hieronymus, G. — (38). Hitchcock, А. S. 1455, 1456 ( 1452, 1458, 1454 (37); — (374). Hood, В. С. — (37); — (37а). Нирег, А. М. 19 (37). Humboldt, A. — (28). Jermy, G. 31, 62, 248, 478 (13). 374 4); en M. — (27). Lindheimer, Е. 111, 325, 504, 652, 677, 1063 (13). Lloyd, F. E. 199 (23). Maxwell, C. F. 49 (13). Mearns, E. A. 1432 (13). Milligan, Mrs. J. M. — (37 Mohr, C. — (37). Nash, G. V. 184, 1774 (37). Nealley, G. C. — (13); 580 (15); 124 (357) (16). Née, D. L. — (24). Nelson, E. W. 4597 (15); 6604 (16). Nicolas, Bro. 299 (24). Niederlein, G. 42 (8). Oersted, A. S. 808 (17). Ohlinger, L. B. 188, 1415, 1437 (37). Orcutt, C. R. 6230 (13). Palmer, Ed. 235, 402 (1); 347 (4); 492 (7); 2083, 9667 (13); 125, 208, 276, 309, 472, 930, 1009, 1010, 1075, 2038 (15); 606 (25); 346 (37a). Palmer, E. J. —, 9708, 9811, 11871, 12229, 13698, 13758, 14252a (18). Parry, С. С. « Palmer, Ed. 699 (15). Patzky — (15). Pieters, А. J. 121 (37). Pringle, С. С. 2154, 2939, 6058 (1); 5481 (11); 66 (15); 6718 (17); 2907, 11318 (18); 6712 (19); 4134 (20); 11322 (24); 5020 (27); 7249, 13838 (34); 4442 (36). Ригриз, С. А. 2362, 3347 (5); 2263 (27). Ravenel, Н. У. — (3 Reverchon, J. —, 722, 1941, 2114 (18). Reynolds, Miss М. С. — (37). Riedel, L. 501 (6); 45 (22). Rolf, P. H. 190 (37a). Rose, J. М. 2259, 3581 (10); 8495 (13); 3615 (15 Rose, J. N. & Rose, J. R. 11148 (15). Rose, J. N., Standley, P. C. & Russell, P. G. 12833, 14298 (1). Rothrock, J. T. —, 637 (15); — (37). Ruiz, L. — (26); — (32). Ruth, A. 30, 81, ps (13). Sartorius, C. — (9). Schaffner, J. G. y Do (15). 2 (9). Seemann, B. 1513 (14). Seler, E. 1733 (1). Sello (Sellow), F. — (29); —, 173 (33); — (380. Short, C. W. — (37). 62 Shreve, Е. 4978 (15). Simpson, J. Н. 528 (4). Small, J. К. & Carter, J. J. 776 (4). Smith, H. H. 2219 (37). Smith, Rev. L. C. p 729 (1); 419 (19). Spruce, R. 5989 (28). Standley, Miss J. P. 13, 179, 443, 12852, 12917 (37а). Standley, P. C. шин (13); 18894 (37a). Тћагр, В. С. 733 (13). Thurber, С. Pos (15); — (37). [Vor. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN Tracy, 5. M. 8079 d 6683 (37). Trelease, W. 352 ( Tweedie, J. 771 даң Tweedy, Е. 180 (13). Venturi, 5. 55 (38). Warming, Е. — (30). Webber, Н. J. 520 (37). Westgate, J. М. 3607 (37). Wilcox, Т. Е. —, 150 (15). Wislizenus, A. 186 (15). Wright, С. 432 (13); 1462, 1463 (15). Young, J. P. 301 (4). INDEX TO SPECIES New genera, species, combinations and forms are printed in bold face type; synonyms in italics, and previously published names in ordinary type. ``. 5,2555 20d. 0 л... ПР 60 ЗАР У 60 нт ........... eere 60 о A УГ 25 И ER TE ІРІ. УУ 54 оо: МОГО 36 а E E 31 Монг, О eo eee 39 о ОНО T 28 ос 22522222. 60 "it 217 io ECT ET E EO E ET а 42 ПРОРОКОТ. 48 ЖЛЕ sie igs bee Pee hoe 30 cubensis A. Rich............... ЦИЛИНОВАИЕ: И ом 39 hirsutissimus . . 28 hi Паје ee Р 51 О ла. ДАРЕ АИ 51 Jasminum-mezicanum ......... 47 д{азапащасвиг................. 32 АЛАШ а си > 86 тататћотав................... 50 ШБИТОРМДИЙН,.........ї 14214 47 oblongifolia D. Шоп........... 55 oblongifolia var. angusta........ 31 oblongifolia var. texensis........ 37 oblongifolius Torr............. 40 ovatus Benth ovatus (Cav.) Nees............ 37 Palmeri АМА. E ен СУ 60 Pulegium 53 quadrangularis ари вани 49 Ир сад у 50 P dM EM мелат 52 AAT Е O 34 Schiedeanus var. multiflorus... .. 34 r EE ес 46 Tweedianus . 59 Піреғасатімы.................. 55 ЮМНЫ... хан ох СІРІ 55 ИКЕ а 86 О": AU. рр 55 ои 25 Р РУО. 54 ‘sical SiN sea er ees мода ин 31 о АЕ 36 о АС 39 DEMNM эз е od И И 60 т . 42 ет 48 шин Е: 41 ПИЗА, кои ИА 30 (OMEN sie ica АГА 60 decumbens................... 39 VOOR ан АЧ cee se 12 НИЛ... Cie a А 60 аа И 222 12, 26 цалин... 2 223: 33 Пр 51 ВИО: Ва аи срива 28 1928] KOBUSKI—MONOGRAPH OF DYSCHORISTE 63 Капи. fox әх. ee 58 беоне... ........... 30 ВИ о. . 60 беруни: ела, 46 hygrophiloides.............. . 51 бидоћаћећета................ 29 заНиселам................. . 35 Tweedbibio, «. XI Е 59 ЈОНИ Е осеке 47 alate ж ДОРИ ан 54 lavandulacea. ................ 32 ОПИШЕ oss а US M DP PEE 60 int. s. 522252222. 36 тепе TUNE. DE TUM 60 PCT. MENTO ca cos E 44 бы... 30 maranhonis Lindau............ 30 ша. Е: аа 26 maranhonis . 50 КҮНГЕ О ere сц <<: а 10 од оса Кеса 47 НОВЕ о... 55 Мийгнин .............-... 94 californica . . 60 €— о ee Ы» 43 n ОЕ 60 оЫорріѓоіа. ................. 55 [с 1 4 ТА ТКТ 48 Kommen glabra.......... 57 о ТОРЕ 10 ИА a рока го ххэ Ч 53 geminiflora var. humilis........ 58 paraguariensis . . 59 ИСА... о rr nnn 47 о eser] e 44 соодо оЙа................. 10, 55 КЕНЕ АНИ И n 42 т f TET ЕЛЕНА 40 УКЕ пасуса Ра 53 МИНИ т А навике 53 e ари ЕЕЕ uS 32 peninsularis. . 60 quadrangularis. . Bow Tm АООТ 50 QUIM. еее 50 БИЕ АВЕ 52 | s © 52 itl. Жар са о арене E 50 T о ра ЕЭ 35 ЕЕ... 60 "rtr. Л es eee 45 Е: 60 М... PERRO TERRE TRUE 1 48 ойр ниа.............-.--.. 50 Schiedeana; 5... о 8: 34 TUPADI Jl RENT RS 50 [У ог. 15, 1928] 64 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 3 Fig. 1. Flower of D. oblongifolia (Michx.) О. Ktze. Fig. 2. Open corolla of e character and position of stam x 8. и (Michx.) 4 Кие. Fig. 3. Open calyx of D. oblongifolta (Michx.) О. Ktze. ХЗ. Fig. 4. Pistil of D. oblongifolia (Michx.) О. Ktze. Fig. 5. Dehiscing capsule of D. oblongifolia (Michx.) О. Ktze. position of retinacula and seeds Х 8. Showing х 8. Showing Fig. 6. Mature capsule (before dehiscence) of D. oblongifolia (Michx.) О. Ktze. Х 3. Showing Ше persistent calyx. PLATE 3 ANN. Мо. Вот. Garb., Vor. 15, 1928 KOBUSKI—MONOGRAPH OF DYSCHORISTE [Vor. 15, 1928] 66 ANNALS OF THE MISSOURI BOTANICAL GARDEN ExPLANATION ОЕ PLATE PLATE 4 Dyschoriste trichanthera Kobuski From the type specimen, Hassler 7780, in the Gray Herbarium of Harvard University. ANN. Mo. Bor. Garp., Vor. 15, 1928 PLATE 4 ча 7755 рони ‹ T р | лла бул hin ^ а. Ant р окт BY ск ковей: КОВОЗКТ—МОХОСВАРН OF DYSCHORISTE [Vor. 15, 1928] 68 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 5 Dyschoriste Purpusii Kobuski From the type specimen, Purpus 2362, in the herbarium of the Missouri Botanical arden. ANN. Mo. Вот. Ganp., Vor. 15, 1928 PLATE 5 DE ] а ы М Ч ! JR Berar: ^ Ad, 4 } 4 % + ot Nag 47 $us KOBUSKI MONOGRAPH OF DYSCHORISTE (У ог. 15, 1928 70 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 6 Dyschorisie Greenmanii Kobuski From the type specimen, Palmer 492, in the United States National Herbarium ANN. Мо. Вот. Garb., Vor. 15, 1928 PLATE 6 UNITED STATES NATIONAL MUSEUM » #44 bd арай ie ФИНО 100 metere та и р НА УИ у 4 4 ! ши: ery ey o d KOBUSKI—MONOGRAPH OF DYSCHORISTE [Уоь. 15, 1928 72 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 7 Fig. 1. Dyschoriste Roset Kobuski From the type specimen, Rose 2259, in the United States National Herbarium Fig. 2. Dyschoriste crenulata Kobuski From the type specimen, Nelson 6604, in the Gray Herbarium of Harvard Univer- PLAT! ANN. Mo. Вот. Garb., Vor. 15, 1928 ф qus «Е. Сп. а а. Le ву с с ко! KOBUSKI—MONOGRAPH ОЕ DYSCHORISTE [У от. 15, 1928] 74 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 8 Dyschoriste jaliscensis Kobuski From the type specimen, Pringle 5481, in the Gray Herbarium of Harvard Univer- sity ANN. Мо. Вот. Garb., Vor. 15, 1928 PLATE 8 О PRINGLE, -Æ МЕХ!САМА. 1803. UE т Юнь нал. аьаа ЕН. 7h 4$ KOBUSKI—MONOGRAPH OF DYSCHORISTE Мог. 15, 1928] 76 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 9 Dyschoriste bilabiata (Seem.) О. Ktze. From the type specimen, Seeman 1513, in the Royal Botanic Gardens at Kew, England. PLATE 9 ANN. Мо. Вот. Garp., Vor. 15, 1928 dm " Ё 929363 Ê) so ISIS ва Ир % - ғ ы кемімен сола (Хам. / 6. er. 1.135 ли менон Bor ARI. өмшин Ao 1513 77 (11-41 ^35, Ї 4. m "y РИД 5 2 : otf, Калалы | Па. /ЕЧР. 4 MONOGRAPH OF DYSCHORISTE KOBUSKI (Мог. 15, 1928] 78 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 10 Dyschoriste oaxacensis Kobuski From the type specimen, Pringle 6712, in the herbarium of the Missouri Botanica! Garden. ANN. Mo. Вот. Garb., Vor. 15, 1928 PLATE 10 + | % A э. Же): \ з ee я ини АҒ. > 2 Ж PRINGLE Р манит а. E ” САМ A | p ен б STATE СР CALAS 6712 Dyschoriste Jasm : 0 "Tm | ЕУ KOBUSKI—MONOGRAPH OF DYSCHORISTE [У он, 15, 1928 80 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 11 Dyschoriste pinetorum Kobuski From the type specimen, Pringle 4134, in the Gray Herbarium of Harvard Uni- versity. ANN. Mo. Вот. Ganp., Vor. 15, 1928 РрАТЕ 11 KOBUSKI—MONOGRAPH OF DYSCHORISTE (Vor. 15, 1928] 82 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 12 Dyschoriste sagittata Kobuski From the type specimen, Fiebrig 6383, in the Gray Herbarium of Harvard Uni- versity. ANN. Мо. Вот. Слво., Vor. 15, 1928 РпАТЕ 12 К. Fiebrig: Plantae paragnayenses JS ИА 7 2 / . 2 0,252 = жесе” 54122) С 6383 Хосеа вад. et. aap pe ae Et: л шээг б à | KOBUSKI—MONOGRAPH OF DYSCHORISTE [Vor. 15, 1928] 84 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 13 Dyschoriste Lloydit Kobuski From the type specimen, Lloyd 199, in the United States National Herbarium. ANN. Mo. Bor. Ganp., Vor. 15, 1928 — wo 499 ЭРЭЛД А Lag ket. hag PLATE 13 KOBUSKI—MONOGRAPH OF DYSCHORISTE [У от. 15, 1928] 86 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 14 Dyschoriste microphylla (Сах.) О. Ktze. From the type specimen, D. Luis Née, in the herbarium of the Botanical Garden, Madrid, Spain. Ann. Мо. Вот. Garp., Vor. 15, 1928 PLATE 14 e М fara] А УУ РТА РРА 1299 0 ay „0. / hady > Son Ў А d А Ecco dé 227 harvi nut upk сын Жалба звал “теу baw.) 444. байс Ила. y ги 237 ds Ка... 7 74: Cag! es KOBUSKI—MONOGRAPH OF DYSCHORISTE (У ог. 15, 1928] 88 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 15 Dyschoriste xylopoda Kobuski From the type specimen, Pringle 4442, in the herbarium of the Missouri Botanical Garden. PLATE 15 ANN. Mo. Bor. Garb., Vor. 15, 1928 Е СТ ТАШ STATE ST АШ НУ Calophanes саран, Erst. KOBUSKI—MONOGRAPH OF DYSCHORISTE [Vor. 15, 1928] 90 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 16. Dyschoriste paraguariensis Kobuski From the type specimen, Hassler 4355, in the Gray Herbarium of Harvard Uni- versity. 1) Ann. Mo. Bor. Garp., Vor. 15, 1928 PLATE 16 Ar 5. НА а i А \ 2 " Мала ААА ҮхААЛУҮХАХ 1 X 1-4 5 >” < ^ ДЕА АЛА. УТУ $. а. KOBUSKI—MONOGRAPH ОК DYSCHORISTE STUDIES IN THE UMBELLIFERAE. Г MILDRED E. MATHIAS Jessie R. Barr Research Fellow in the Henry Shaw School of Botany of Washington University A critical study of the genus Cymopterus has necessitated a detailed investigation of about twenty allied genera including Glehnia Schmidt. Asa Gray in 1859 doubtfully referred a plant from the Cooper collection in the region of Puget Sound, Washington, to the genus Cymopterus, and in 1860 published the species as Cymopterus 7 littoralis. Bentham (’67) in Bentham and Hooker’s ‘Genera Plantarum,’ published in September, 1867, transferred this species to his new genus Phellopterus. Е. Schmidt (67), some time between January and July inclusive of the year 1867, in his ‘Prolusio Florae Japonicae’ published the new genus Glehnia? with one species, G. littoralis, basing it on à Maximowiez plant from Hakodate. In this work Schmidt also included Cymopterus ? littoralis Gray as a distinet species. In 1868 in his 'Flora Sachalinensis’ he recognized that his Glehnia littoralis was con- specific with Cymopterus 7 littoralis Gray and adopted the generic name Phellopterus of Bentham. Upon critical examination of the Cooper and Maximowiez types Schmidt's view as to the congenerie nature of the two is confirmed. Аз the generic name Glehnia of Schmidt was published at least two months prior to the Phellopterus of Bentham, on the basis of priority, it must be retained as the correct name for the genus and the Maximowicz plant must be taken as the generic type. The historical type of the genus is then the plant collected by Maximowiez in Hakodate, Japan, in 1861 and must bear the specific name Glehnia littoralis chmidt. Bentham, С. ('67)) In Bentham, G., and J. D. Hooker, Genera Plantarum 1: 5. September, 1867. Gray, Asa (759). “Botany of Japan." Mem. Am. Acad. N.S. 6°: 391, 428. 1859. ------, 060). Stevens’ Report of U.S. Explorations & Surveys from the Mis- sissippi River to the Pacific Ocean 122: 62. 1860. 1 Issued April 30, 1928. ? The genus Glehnia was so named in honor of Peter von Glehn who collected with Schmidt on the Island of Sachalin. ANN. Mo. Bor. Garp., Vor. 15, 1923 (91) ГУ оз. 15 92 ANNALS OF THE MISSOURI BOTANICAL GARDEN Schmidt, Е. (67). Prolusio Florae Јаропісае іп Ма. Ann. Mus. Bot. Lugd. Bat. 3: 61. January-July, 1867; Prolusio Florae Japonicae, 249. 1867. ‚ (68). Flora Sachalinensis. Mem. Acad. Imp. Sci. St. Petersbourg, УП, 12: 138-140. 1868 Glehnia Schmidt, Prol. Fl. Jap. in Miq. Ann. Mus. Bot. Lugd. Bat. 3: 61. Jan.-July, 1867; Prol. Fl. Jap. 249. 1867; Baillon, Hist. Plant. 7: 215. 1880; Coult. & Rose, Contr. U. 5. Nat. Herb. 7: 165. 1900; Piper, Contr. U. 5. Nat. Herb. 11: 429. 1906; Henry, Fl. S. Brit. Col. 223. 1915; Piper & Beattie, Fl. М. W. Coast, 267. 1915; Carter & Newcombe, Prel. Cat. FI. Vanc. 61. 1921. Phellopterus Benth. in Benth. & Hook. Gen. Pl. 1: 905. Sep- tember, 1867, not Phellopterus Nutt. (section under Cymopterus in Torr. & Gray, Fl. N. Am. 1: 623. 1840) іп Соц. & Rose, Contr. U. S. Nat. Herb. 7:166. 1900; Schmidt, Mem. Acad. Imp. Sci. St. Petersbourg, УП, 122: 138. 1868; Franchet & Savatier, Enum. Plant. Jap. 1: 185. 1875; Wats. Bibl. Ind. 1: 430. 1878; Franchet, Cat. Plantes, in Mem. Soc. Nat. Sci. Cherbourg 24: 221. 1884; Coult. & Rose, Rev. N. Am. Umbell. 21, 81. 1888; Macoun, Check List Can. Plants, 25. 1889; Cat. Can. Plants 5: 329. 1890; Howell, Е. М. W. Am. 1: 259. 1898; Engl. & Prantl, Nat. Pflanzenfam. 3*: 221. 1898; Ito & Matsumura, Tent. Fl. Lutch. in Jour. Coll. Sci. Imp. Univ. Tokyo 12: 529. 1899; Yabe, Rev. Umb. Jap. in Ibid. 162: 92. 1902; Boiss. Omb. Cor. in Bull. Herb. Boiss. II, 3: 955. 1903; Nakai, Fl. Kor. 1 in Jour. Coll. Sci. Imp. Univ. Tokyo 26!: 272. 1909. Herbaceous, subacaulescent, glabrous or pubescent perennials. Leaves coriaceous, petioled, bipinnatisect, broadly ovate in general outline. Inflorescence pedunculate, villous, peduncles shorter than or equalling the leaves; involucre usually absent, sometimes present in the form of a few linear bracts; involucel of conspicuous linear-lanceolate bracts. Calyx teeth inconspicuous. Stylopodium lacking. Fruit ovate-oblong to globose, glabrous or pubescent, flattened dorsally; lateral and dorsal wings present; wings broadened at the base; oil-tubes large, numerous, 2–6 on the commissural side; strengthening cells absent. Type species: Glehnia littoralis Schmidt, Prol. Fl. Jap. in Miq. Ann. Mus. Bot. Lugd. Bat. 3: 61. 1867; Prol. Fl. Jap. 249. 1867. 1928) MATHIAS—STUDIES IN THE UMBELLIFERAE. I 93 ABBREVIATIONS The following abbreviations have been used in citations to indicate the different herbaria from which material has been obtained for study: M = Missouri Botanical Garden Herbarium; С = Gray Her- barium of Harvard University; NY = New York Botanical Garden Herbarium; US = United States National Herbarium; W = Herbarium of the University of Washington deposited in the Washington State Museum; О = Herbarium of the Univer- sity of Oregon; OAC - Herbarium of the Oregon Agricultural College; C = Herbarium of the University of California; P = Herbarium of Pomona College. Ккү TO THE SPECIES Fruit pubescent, species of the eastern hemisphere................ 1. G. 203 Fruit essentially glabrous, species of the western ћепиврћеге........ 2. G. leio 1. Glehnia littoralis! Schmidt, Prol. Fl. Jap. in Miq. Ann. Mus. Bot. Lugd. Bat. 3: 61. 1867; Prol. Fl. Jap. 249. 1867. PL 17, Ge, Е: 18; PL 19, fig. 1. “ Archangelica officinalis, Hoffm.?" in Gray, “ Account of the Botanical Specimens" from Narrative of the Perry Expedition 2: 312. 1856. “Суторегиз (2) littoralis, glaber" Gray, “Botany of Japan," in Mem. Am. Acad. Х. 8. 62: 428. 1859, nomen nudum. Cymopterus ? littoralis Gray, "Botany of Japan" іп Mem. Am. Acad. N. S. 67%: 391, 428. 1859, as to specimens from eastern hemisphere, nomen nudum. “Суторетв glaber (A. Gray)" Black, "Catalogue of Japan 1Glehnia littoralis Schmidt, em.—Planta humila, subacaula; foliis, petiolis excludentis, 5-13 cm. longis latisque, supra hirtellis in rachides nervosque, subtus glabris vel crebre tomentosis, ultimis segmentis foliorum oblongo-obovatis vel segmentis terminalibus cuneatis, 0.5-5 cm. longis, 0.4-4 cm. latis, apice rotundatis vel acutis, plus minusve ipso cie i petiolis 3-12 em. longis, subdilatatis, hirtellis vel glabris; umbellat tis, crebre villosis; pedunculis subcrassis, subinde ramosis, foliis bre ae vel aequantibus; umbellis patulis, 6-30-radiatis, radiis 1-3.5 em. longis; involucro 1-3-bracteato; umbellulis capitatis, bracteis involucellorum pluribus, lanceolato-attenuatis; fructibus ovato-oblongis vel subglobosis, 0.4-1.5 cm. longis, villoso-pubescentibus, pilis Wii i alis lateralibus saepe dorsalibus latioribus; vittis multis, 2-6 in commissurem —Col- lected in Hakodate, Japan, 1861, Mazimowicz (Gray Herb.), С0-ТУРЕ. (Мог. 15 94 ANNALS OF THE MISSOURI BOTANICAL GARDEN Plants” in Hodgson, “А residence at Nagasaki and Hakodate in 1859-1860,” 335. 1861, nomen nudum; Bonplandia 10: 92. 1862, nomen nudum. Phellopterus littoralis (Gray) Benth. in Benth. & Hook. Gen. РІ, 1: 905. 1867, as to plants of eastern hemisphere; Hance, Spic. Е. Sin. in Jour. Bot. 16: 12. 1878; Forbes & Hemsley, Jour. Linn. Soc. Bot. 23; 331. 1888; Engl. & Prantl, Nat. Pflanzen- fam. 38: 221. 1898, as to plants of eastern hemisphere; Ito & Matsumura, Tent. Fl. Lutch. in Jour. Coll. Sci. Imp. Univ. Tokyo 12: 529. 1899; Yabe, Rev. Umb. Jap. in Ibid. 16?: 93. 1902; Boiss. Omb. Cor. in Bull. Herb. Boiss. II, 3: 955. 1908; Nakai, Fl. Kor. 1. in Jour. Coll. Sci. Imp. Univ. Tokyo 26!: 272. 1909; Fl. Kor. 2. in Ibid. 31: 492. 1911. “С. glaber" Gray acc. to Schmidt, Mem. Acad. Imp. Sci. St. Petersbourg, УП, 127: 139. 1868, nomen nudum. “ Phellopterus littoralis” ace. to Schmidt, Mem. Acad. Imp. Sci. St. Petersbourg, УП, 12?: 138. 1868. ** Phellopterus littoralis Schmidt" acc. to Franchet & Savatier, Enum. Plant. Jap. 1: 185. 1875; Franchet, Cat. Plantes, in Mem. Soc. Nat. Sci. Cherbourg 24: 221. 1884. “Glehnia littoralis (Gray) Schmidt" acc. to Соц. & Rose, Contr. U. 5. Nat. Herb. 7: 165. 1900, as to plants of eastern hemisphere. Low subacaulescent plants; leaves, excluding petiole, 5-13 cm. long, about as broad, hirtellous on the rachises and nerves of the upper surface, glabrous to densely tomentose beneath, the ul- timate leaf-segments oblong-obovate or the terminal segments cuneate, 0.5-5 cm. long, 0.4-4 em. broad, rounded to acute at the apex, somewhat unequally cartilaginously dentate; petioles 3-12 cm. long, somewhat inflated, hirtellous to glabrous; in- florescence pedunculate, densely villous; peduncles stoutish, sometimes branched, shorter than or equalling the leaves; umbels spreading, 6-30-гауед, rays 1-3.5 cm. long; involucre 1-3-bracted ; umbellets capitate, bracts of the involucel several, lance-attenu- ate; fruit ovate-oblong to subglobose, 0.4-1.5 em. long, villous- pubescent with multicellular hairs, lateral wings usually broader than the dorsal wings; oil-tubes numerous, 2-6 on the com- missural surface. 1928] MATHIAS—STUDIES IN THE UMBELLIFERAE. I 95 Type specimen: Mazimowicz, ‘‘Glehnia littoralis Е. Schmidt. Fl. Sachalin ined.” Iter secundum. Japonia. Hakodate. 1861. (ТУРЕ probably in Herb. Leningrad; со-түрЕ in the Gray Her- barium of Harvard University). Distribution: eastern hemisphere, along sandy sea-shores, from southern China northward, and in Japan. This plant is commonly known in Japan as '' Hama-bofu" in relation to its maritime habitat, hama meaning sea-coast, and bofu, a medicinal plant. Specimens examined: JAPAN: Insula Sachalin, 1860, Schmidt (С); Kamiiso, Prov. Oshima, Hokkaida, 12 July, 1890, Miyabe & Tokubuchi (С); Hakodate, Iter secundum, 1861, Mazximowicz (G co-TYPE); Insula Jesso, circa Hakodate, 1861, Albrecht (С); Yezo, Ishikari, 10 Sept. 1903, Arimoto (С, М); Nambu, Nippon, 1865, Mazimo- wicz, coll. Tschonoski (NY); Isoya, Shiribeshi, July, 1883, Таке- nobu (G); seashore, Prov. Rikuzen, 9 July, 1913, Yasuda (W); Isl. Futami, 24 June, 1910, Flora Japonica, collector unknown (US 1155343); Loo-Choo Islands, 1853-56, Wright 98 (G, US); Corea, 1859, Wilford (NY). SIBERIA: Vladivostok and vicinity, May-Oct. 1919, Topping 2236 (G). СнтмаА: Tsingtao, 1911, Zimmermann (С, US 795348); ''Putoo Island—Clekiane," Henry (M); “Рообоо Isle, Chekiang," Faber М (US); Delatache and Amoy, Henry (NY). 2. Glehnia leiocarpa! Mathias, nom. nov. Pl. 17, fig. 1, 4; Pl. 19, fig. 2. Cymopterus ? littoralis Gray, Mem. Am. Acad. N. 5. 6?: 391, 428. 1859, as to American specimens, nomen nudum; Stevens' Rept. U. S. Expl. & Surv. from Miss. to Расте Ocean 12: 62. 1860; Jeps. Man. Fl. Plants Calif. 731. 1925. Phellopterus littoralis (Gray) Benth. in Benth. & Hook. Gen. г Glehnia leiocarpa Mathias, nom. nov.—Planta humila, subacaula; foliis, petiolis excludentis, 2.5-15 ст. longis latisque, supra hirtellis in rachides nervosque, subtus crebre tomentosis, ultimis segmentis foliorum oblongo-obovatis vel segmentis ter- minalibus cuneatis, 0.5-5 cm. longis, 0.4-3 сіп. latis, apice rotundatis vel acutis, lus minusve dentatis, marginibus subinde cartilaginibus; petiolis 2.5-14 ст. longis, subdilatatis, hirtellis; inflorescentiis umbellatis pedunculatis, crebre villosis; pedun- culis suberassis, subinde ramosis, saepe foliis brevioribus, rare aequantibus; umbellis [Vor. 15 96 ANNALS OF THE MISSOURI BOTANICAL GARDEN Pl. 1: 905. 1867, as to American plants; Engl. & Prantl, Nat. Pflanzenfam. 38: 221. 1898, as to American plants. ** Glehnia littoralis (Gray) Schmidt" асс. to Coult. & Rose, Contr. U. S. Nat. Herb. 7: 165. 1900; Piper, Contr. U. S. Nat. Herb. 11: 429. 1906; Piper & Beattie, Fl. N. W. Coast, 267. 1915; Carter & Newcombe, Prel. Cat. Fl. Vane. 61. 1921. “ Phellopterus littoralis Schmidt" асс. to Wats. Bibl. Ind. 1: 430. 1878; Coult. & Rose, Rev. N. Am. Umbell. 81. 1888; Macoun, Check List Can. Plants, 25. 1889; Cat. Can. Plants 5:329. 1890; Howell, Fl. N. W. Am. 1: 259. 1898. Glehnia littoralis Schmidt асс. to Henry, Fl. S. Brit. Col. 223. 1915. Low subacaulescent plants; leaves, excluding petiole, 2.5-15 em. long, about as broad, hirtellous on the rachises and nerves of the upper surface, mostly densely tomentose beneath, the ultimate leaf-segments oblong-obovate or the terminal segments cuneate, 0.5-5 ст. long, 0.4-3 ст. broad, rounded to acute at the apex, unequally dentate, margins sometimes cartilaginous; petioles 2.5-14 em. long, somewhat inflated, hirtellous; inflores- cence pedunculate, densely villous; peduncles stoutish, sometimes branched, usually shorter than the leaves, rarely equalling them; umbel globose to spreading, 5-13-rayed, rays 0.5-4.5 ст. long; involucre 1—3-bracted; umbellets capitate, bracts of the involucel several, lance-attenuate; fruit ovate-oblong to subglobose, 0.4–1.2 cm. long, essentially glabrous (sometimes with a few scattered multicellular hairs), lateral wings sometimes broader than dorsal wings; oil-tubes numerous, 2-6 on commissural surface. Type specimen: J. G. Cooper, ‘‘sandy shores, Washington Terr. (Shoal Water Вау).” 1854. (The type is in the Gray Herbarium of Harvard University and is labeled ‘‘Cymopterus ? littoralis, n. sp." in Gray's handwriting; co-types are in the Herbarium of the New York Botanical Garden and in the United States National Herbarium.) globosis vel patulis, 5-13-radiatis, radiis 0.5-4.5 em. longis; involucro 1-3-bracteato; umbellulis capitatis, bracteis involucellorum pluribus, lanceolato-attenuatis; fructibus ovato-oblongis vel subglobosis, 0.4-1.2 cm. longis, fere glabris vel subinde sparse pubescentibus, pilis multicellulatis; alis lateralibus dorsalibus latioribus; vittis mul- tis, 2-6 in commissurem.— Collected on sandy shores, Shoal Water Bay, Washington Territory (State of Washington), 1854, J. С. Cooper (Gray Herb.), TYPE. 1928] MATHIAS—STUDIES IN THE UMBELLIFERAE, I 97 Distribution: North America along sandy sea-coasts from San Francisco, California northward. Specimens examined: ALASKA: along the Ankow River, near Ocean Cape, vicinity of Yakutat Bay, 1 July, 1892, Funston 51 (NY, M, C British CoLuMBIA: vicinity of Ucleulet, Long Beach, Van- couver Island, 25 June, 1909, Macoun 78600 (US); sand, Oak Bay, Vancouver Island, 31 May, 1887, Macoun (G). WasHINGTON: Lopez, San Juan Islands, 25 June-1 Aug. 1917, S. M. & E. B. Zeller 963 (NY, M, G, US); Puget Sound, Wilkes Expedition (NY, US 44092); Port Angeles, 26 June, 1908, Webster (W); sand dunes, Ocean Park, April, 1908, Rigg (УУ): Ilwaco, 21 June, 1904, Piper 5002 (US); Oyhut, Chehalis County, 7 June, 1897, Lamb 1249 (NY, M); drifting sand, common along the ocean beach, Westport, Chehalis Co., 26 June, 1892, Hender- son 885 (US); ocean beach, Westport, Chehalis County, 26 June, 1892, Henderson (W); sand dunes, Westport, June, 1917, Grant (NY); sand spit, Sequim, June, 1915, Grant (NY, М 788926); Seattle, July, 1915, Freiberg (M. 813695); sandy dunes, mouth of “Joe Creek," near Moclips, 28 June, 1908, Foster 824 (US); sandy sea-shores, Port Angeles, 26 June, 1908, Flett 3375 (US); Olympie Mts., Clallam Co., July, 1900, Elmer 2768 (NY, M, US); M. Beach, Westport, 10 July, 1907, Cowles 512 (M); “sandy shores, Washington Terr. (Shoal Water Bay)." 1854, Cooper (а түре, NY, US); beach sand, Copalis, June-July, 1902, Conrad 392 (05), Copalis, 30 May, 1912, Bardell (M. 813656). OnEGON: Clatsop Beach, Clatsop Co., 21 Aug. 1902, Sheldon 11252 (NY, M, G, P, US); Gearhart, 19 June, 1904, Piper 6241 (US); Gearhart, 19 June, 1904, Piper 6131 (US); sandy sea- beach, Newport, 3 July, 1918, J. Nelson 2292 (С); Nestart’s Bay, Tilamook Co., 29 June, 1894, Lloyd (NY); on strand, Nestucca, July-Aug. 1901, Kirkwood 149 (NY); sandy sea-shore, mouth of the Umpqua River, 18 June, 1885, Howell! (OAC, US, M, 1151); on sand dunes, mouth of Tillamook Bay, 16 July, 1882, Т. Howell (NY); on shifting sand, Tillamook Bay, 14 July, ‘Thomas Howell in the earlier period of his botanical career used the signature Thomas J. Howell which accounts for the discrepancy in names appearing on herbarium labels. [Vor. 15 98 ANNALS OF THE MISSOURI BOTANICAL GARDEN 1882, Т. J. Howell (М, US 33339); Clatsop Beach, 26 July, 1891, J. Howell (M 863104); on shifting sands of sea-shore, Coos Bay, 19 Aug. 1911, House 4705 (US, NY); drifting sand, ocean beach, Tillamook Bay, 14 July, 1882, Howell & Henderson (0); drifting sand, ocean beach, Clatsop, 30 July, 1887, Henderson 385 (M, OAC); beach, below Florence, 20 May, 1925, Henderson (O); sand of the ocean above high tide, Rockaway, 16 Sept. 1925, Henderson (O); sea-shore, Fort Stevens, 7 July, 1886, Henderson (O); Bayocean, Garibaldi, 28 Aug. 1914, Hitchcock 12370 (US); sands of the Oregon coast between Umpqua and Coos Вау, 12 Aug. 1880, G. Engelmann (M); on sand dunes of the ocean, Gearhart, Clatsop County, 1 Sept. 1898, Coville 861 (US). CALIFORNIA: in drifting sand, Humboldt County, sand hills of ocean beach at Samoa, opp. Eureka, 7 Aug. 1901, Tracy 1261 (C); Samoa Beach, Humboldt Co., 17 June, 1911, Smith 3854 (NY); Trinidad, Humboldt Co., 7 June, 1911, Smith 3806 (NY); Trini- dad, Humboldt County, 6 July, 1911, Smith 3806 (US); Pebble Beach, Crescent, Del Norte Co., 17-20 June, 1925, Parks 8257 (C 279023); sandy dunes at Humboldt County camp, 7 miles south of Trinidad, 24 July, 1924, A. A. Heller 13882 (NY); Crescent City, Del Norte Co., 30 June, 1899, Davy & Blasdale 5960 (C); Point Arena, Mendocino Co., 24 July, 1900, Davy 6050 (C); peninsula, Eureka, 23 Aug. 1904, Congdon (C 140694); sea-shore peninsula, Eureka, Humboldt Co., 23 July, 1904, Congdon (M); Humboldt Bay, May, 1901, Chandler 1145 (C); Trinidad, Humboldt Co., 18 July, 1916, Abrams 6140 (NY, O). The genus Glehnia is characterized by its maritime habitat, broad leaf divisions, thick coriaceous texture of the leaves, and prominent wing development of the fruit. 'The two species are separated largely on fruit characters. Glehnia littoralis Schmidt, the species of the eastern hemisphere, always has a pubescent fruit. The pubescence is villous with multicellular hairs. The mature fruit may be only slightly pubescent due to the falling off of the hairs but in such cases it has a tuberculate appearance showing the previous attachment of these hairs. In the young fruit the pubescence is densely villous. As a rule the oil-tubes of the fruit are smaller and more numerous than in the other 1928] MATHIAS—STUDIES IN THE UMBELLIFERAE. I 99 species. The characters of inflorescence and foliage are similar in both species. There is quite a range of variation in foliage pubescence of Glehnia littoralis. The type of the species, the Maximowiez plant from Hakodate, represents an intermediate condition, and upon an examination of additional material may prove to be a hybrid between the glabrous and pubescent forms (pl. 18, fig. 2). The leaves are hirtellous on the lower surface and on the veins and rachises of the upper surface. The one extreme of variation in pubescence is typified by the plant col- lected by Wright in the Loo Choo Islands and labeled by Gray "^ Cymopterus littoralis 77 Gray, var. glabra, vel sp. aff." (pl. 18, fig. 1). The leaf is essentially glabrous, the hirtellous condition being limited entirely to the veins and rachises. The margins of the leaves are more frequently cartilaginous than in other forms. The other extreme of variation is typified by the plant collected by Schmidt in Sachalin in 1860 (pl. 19, fig. 1). This plant super- ficially more closely approaches the species of North America. The lower surface of the leaf has the same dense tomentose pubescence that occurs in Glehnia leiocarpa. However, an ex- amination of a large amount of material from the eastern hemi- sphere shows a great number of intergrading forms; a gradual variation exists from the extreme glabrous form to the densely tomentose one. The fruit in all forms is similar, and the pubes- cence characters of the foliage are of no value in separating Glehnia littoralis into varieties or forms. Glehnia leiocarpa, on the other hand, shows a very constant pubescence character. The leaves in every case are densely tomentose beneath. The fruit is glabrous with the exception of occasional multicellular hairs on the margins of the wings. In no case was а tuberculate appearance observed which would point to the previous attachment of hairs in younger conditions. The young fruit in most cases is essentially glabrous. Moreover, a cross-section of the fruit shows the oil-tubes to be larger and generally fewer in number than in С. littoralis. An interesting geographical distribution is shown in connection with this genus. The two closely related species occur along the coast on both sides of the Pacific Ocean (fig. 1). Glehnia leiocarpa extends from Alaska to northern California and G. littoralis from [Уоь. 15 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN 100 “втацеро Jo seroeds jo поцпапдвтр Зшмочв 1214 a XILOHHO i qp pm а n бет m «өл P qe мој «зол 15 ауз] ruya wiog үчээтэ qn 4 511922441] 2144210 222 ФАК сүзе 9144210 = ө4хөөөгө| елеҹә) us 13231 1928] MATHIAS—STUDIES IN THE UMBELLIFERAE. I 101 Siberia to southern China and through Japan. It is also inter- esting to note the distribution of the different pubescence types of G. littoralis. The more pubescent plants and those most nearly approaching G. leiocarpa occur in the northern region of the distribution area of the species, while the more glabrous plants are found in the southern range of distribution. Such a dis- tribution seems to indicate that the ancestors of this species occurred in the intermediate area and in the land bridge con- necting North America and Asia somewhere in the Bering Sea region. A similar distribution for other genera has been pointed out by various workers in this field. One of the earliest important works was Dr. Gray’s! article on the ‘Botany of Japan" in which he showed the similarity of the flora of northwest as well as eastern America to that of Japan. In this work he also mentions the distribution of the genus Glehnia. Butters,? more recently, has pointed out a similar distribution for the genus Athyrium. Berry? has shown this distribution for Castanopsis, Разатла, Corylus, Juglans, and other genera. The writer is indebted to Dr. George T. Moore, Director of the Missouri Botanical Garden, for the use of the library and ћег- barium of that institution. Sincere appreciation is due Dr. N. L. Britton and Dr. J. K. Small of the New York Botanical Garden, Dr. B. L. Robinson and Dr. Ivan M. Johnston of the Gray Herbarium, Dr. Wm. R. Maxon of the United States National Herbarium, Prof. L. F. Henderson of the University of Oregon, Dr. Helen M. Gilkey of Oregon Agricultural College, Prof. T. C. Frye and Miss Martha R. Flahaut of the University of Washing- ton, Dr. N. L. Gardner of the University of California, and Dr. Philip A. Munz of Pomona College for the privilege of examining material in the herbaria of the above-mentioned institutions or for the loan of material necessary for this study. Thanks are also due Dr. John H. Barnhart of the New York Botanical 1 Gray, A. "Botany of Japan." Mem. Am. Acad. N.S. 62: 376-449. 1859. 2 Butters, Е. К. Taxonomic and geographic studies in North American ferns. I. The genus Athyrium and the North American ferns allied to Athyrium Filix- femina. Rhodora 19: 169-207. 1917. 3 Berry, Е. W. Tree ancestors. 270 рр. 1923. Мог, 15 102 ANNALS OF THE MISSOURI BOTANICAL GARDEN Garden, Dr. J. N. Rose of the United States National Herbarium, and Dr. F. A. F. C. Went of the Botanical Laboratory of Utrecht for their assistance in bibliographical details. Especial thanks are due Dr. J. M. Greenman, Curator of the Herbarium of the Missouri Botanical Garden, for his advice and assistance. List oF ExsIccATAE The distribution numbers are printed in italics. The number in parenthesis is the species number used in this revision. Abrams, L. 6140 (2). Howell, J. — (2). Albrecht, T — (1). Howell, T. — (2). Arimoto, S. — (1). Howell, T. J. — (2). Bardell, т М. — (2). Kirkwood, J. Е. 149 (2). Chandler, H. P. 1145 (2). Lamb, F. H. 1249 (2). Congdon, J. W. — (2). Lloyd, F. E. — (2). Conrad, H. В. 392 (2) Macoun, J. —, 78600 (2) Cooper, J. G. — (2). Maximowicz, C. J. — (1). Coville, F. V. 861 (2). Maximowicz, C. J. (coll. Tschonoski) — Cowles, Н. С. 512 (2). 1). Davy, J. B. 6050 (2). Miyabe, K. and Tokubuchi, E. — (1). Davy, J. В. and Blasdale, У. С. 5960 Nelson, J. С. 2292 (2). (2). Parks, Н. Е. 8957 (2). Elmer, A. D. Е. 2768 (2). Piper, С. У. 5008, 6131, 6241 (2). Engelmann, С. — (2). Rigg, С. В. — (2). Faber, E. M® (1). Schmidt, F. — (1). Flett, J. B. 3376 (2). Sheldon, E. P. 11252 (2). Fl. Japonica (collector unknown), — (1). Smith, H. H. 3806, 3854 (2). Foster, A. В. 824 (2). Takenobu, 8. — (1). Freiberg, G. W. — (2). Topping, L. ү (1). Funston, F. 51 (2). Тгасу, 4. Р. 1261 (2). Grant, J. M. — (2). Webster, E. B. — (2). Heller, A. A. 18882 (2). Wilford, C. — (1). Henderson, L. Е. — 385 (2). Wilkes Expedition, — (2). Henry, A. — (1). Wright, C. 98 (1). Hitchcock, A. 5 12370 (2). Yasuda, А. — (1). ouse, Н. 1. 4705 (2). Zeller, 5. М. апа Е. В. 968 (2). Howell Т. 22 Henderson, L. Е. — (2). Zimmermann, В. — (1). 1928 MATHIAS—STUDIES IN THE UMBELLIFERAE. I 103 INDEX OF SPECIES New species and combinations are printed in bold face type; synonyms in tialics; and previously published names in ordinary type. “ Archangelica officinalis Hoffm.?" 93 101 vU gabor СЕКУ о са ТУТ 94 "piri an Ано Арне гара 101 ҚА и ЕУ бы ‚ 92 Cymopterus ofa Phellopterus Nutt. 92 Cymopterus? littoralis Gray.. .91, 93, 95 42. littoralis?? Gray var. 4 « « « оф Glehnia Schmidt. ........ 91, 98, 101 Glehnia leiocarpa Mathias Li ы она ши 93, 95, 99, 101 Glehnia littoralis Schmidt А 91, 92, 98, 95, 96, 98, 99, 101 РЯ 94, 96 Т 55. Ган ои ико 101 Рава ша, су Уа со анале» а 101 Phellopierus 4, „ЖАЛЫМ ЗУУ Ee 91,92 Phellopterus Nutt ............. 92 Phellopterus и (Gray) Benth. MOIS a NI ЕЛЕ таван 94, “ Phellopterus littoralis Schmidt ” ~ 96 * Phellopterus littoralis” acc. тл ОВС corer ey ee 94 [Vor. 15, 1928] 104 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 17 . Mature fruit of Серта leiocarpa Mathias, collected on “sandy shores, Washington Terr. (Shoal Water Bay)," Cooper, 1854 (Gray Herb.), туре. X 6. Fig. 2. Mature fruit of Glehnia littoralis Schmidt, collected on the Island of Sachalin, Schmidt, 1860 (Gray Herb.). Fig. 3. Mature fruit of Glehnia litoralis Schmidt, collected in Yezo, Ishikari, Arimoto, 10 Sept. 1903 (Mo. Bot. Gard. Herb.). X 6. Fig. 4. Cross-section in median plane “of immature fruit of Glehnia leiocarpa Mathias, collected on “зап4у shores, Washington Terr. (Shoal Water Bay)," Cooper, 1854 (Gray a ), TYPE. Fig. 5. ss-section in мэм plane of mature fruit of Glehnia littoralis Schmidt, collected on is Island of Sachalin, Schmidt, 1860 (Gray Herb.). х 10. — "е ANN. Mo. Bor. Garp., Vou. 15, 1928 PLATE 17 MATHIAS—STUDIES IN UMBELLIFERAE [Vor. 15, 1928] 106 ANNALS OF THE MISSOURI BOTANICAL GARDEN ExPLANATION OF PLATE PLATE 18 Glehnia litioralis Schmidt Fig. From specimens in the Gray Herbarium of Harvard University, namely Wright and Albrecht. Fi From co-type specimen, Mazimowicz, in the Gray Herbarium of Harvard Universit PLATE 18 8 F VoL. 15, 19 ARD., А J Амм. Мо. Вот. С Ex herb. borti bet. Petropolitani. Махинончсз. lier secundam ftp LE hs Зарев. latis Heise, circa Неше De Арғы 5а ЖҰ кұм матасы Japonia. Hakodate. MATHIAS—STUDIES IN UMBELLIFERAE ГҮ ог. 15, 1928 108 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 19 Fig. 1. Glehnia littoralis Schmidt, tay a specimen collected by Schmidt, in the Gray Herbarium of Harvard Univers ig. 2. Glehnia leiocarpa Mathias, а Ше буре вресітеп, Соорет, іп Ше Сгау Herbarium of Harvard University. 19 PLATE Вот. GARD., Vor. Mo. ANN. . Ca a лы Долене. po да, , - e ^ Md MATHIAS -STUDIES IN UMBELLIFERAE CONCERNING THE STATUS OF THE GENUS LATERNEA DAVID H. LINDER Mycologist to the Missouri Botanical Garden Instructor in the Henry Shaw School of Botany of Washington University While in Cuba during the summer of 1924, the writer collected a member of the Clathreae which was subsequently determined in Saccardo (’88) as Clathrus triscapus (Turpin) Fr. In going over the literature concerning the simple columnar species of Clathrus, it was observed that there were few statements as to the manner in which the gleba is borne. Examination of the figures accompanying the original descriptions led the writer to the conclusion that the majority of these simple species carry the gleba in the same fashion as do the more complex ones in which the columns anastomose to form a latticed sphere. In these latter forms, exemplified by Clathrus cancellatus and C. crispus, the gleba is closely applied to the inside of the columns or receptacles. However, in the genus Laternea, of which L. triscapa is the original species, the columns, strictly speaking, are stipes united above, and these subtend an angular body, subovate in outline, the “lanterne” of Turpin (1822). A comparison of C. columnatus, C. crispus, and C. cancellatus brings out the fact that except for the gross morphological dif- ferences, the structure of the simple columnar and the more complex latticed species is essentially the same; that is, the columns may be relatively rough or even smooth on the outside, but on the inside they are always rough and pitted (pl. 20, figs. 3-6). It is to this pitted inside surface of the columns that the gleba is applied. Studies of preserved young material of Clathrus columnatus and observations of the other two species in the field at all stages of development amply support this view. There is certainly no evidence that, at the time of rupture of the volva, any definite receptacle other than the column is present. Aside from the fact that the gleba of Laternea triscapa is strictly confined to the angled, subovate, specialized receptacle pendant from the junction of the apices of the columns, it differs from Ann. Мо. Вот. Garp., Vor. 15, 1928 (109) (Von, 15 110 ANNALS OF THE MISSOURI BOTANICAL GARDEN С. columnatus and other similar members of that genus by being proportionately taller and more slender. In addition, the col- umns are less angular and the surfaces are smooth, both on the inside and outside (pl. 20, fig. 1, 2). With the above distinction in mind, it becomes quite evident that Turpin (1822) was thoroughly justified in creating the genus Laternea for that form which bears the gleba in the manner men- tioned. Accepting this view, then Laternea triscapa becomes the only representative of the genus and Laternea columnata, L. pusilla, L. rhacoides, L. Spegazzini, L. angolensis, and possibly L. bicolumnata, considered as belonging to the genus by Lloyd (09), should be excluded and treated as members of the genus Clathrus, following the treatment by Fischer (86). Certainly this is a more natural grouping, especially since Clathrus columnatus tends towards the more complex type represented by C. cancel- latus. Тһе latter species may at times be columnar below and only show anastomosis of the receptacle above, while in the former, as is shown in pl. 20, fig. 6, there is a tendency for the columns to divide to produce four or even five. If, however, it is deemed more convenient to separate the simple columnar members from the genus Clathrus, then the genus Colonnaria Rafinesque (1808), on the basis of priority, should be restored, and Clathrus of Michelius (1729) should be reserved for those forms with anastomosed receptacles. | In view of the former uncertainty concerning Laternea, it seems advisable, while restoring it to its original status, to ге- describe the genus, and also the species as follows: Laternea Turpin: Columns slender, smooth, usually three, subtending from the junction of the apices an angular, subovate receptacle to which the gleba is restricted. Laternea triscapa Turpin: columns 3, “сарисше Бий”! at base, becoming ‘‘cadmium orange" above; smooth on inner and outer surfaces, 5-6.2 cm. long, 4-5 х 6 mm. in diameter, united above; receptacle pendant, “пора! red," angled, subovate in outline, 10 х 13 mm.; gleba deep olive; volva white, 15 x 20 mm. In sugar cane field at edge of woods, Soledad, Cuba. бер- 1 Ridgway, В. Color standards and nomenclature. Washington, D. C., 1912. 1928] ТЛМРЕН— $Т/10$ OF THE GENUS ГАТЕВМЕА 111 tember, 1924, Linder (in Farlow Herb. at Harvard Univ. and writer’s herbarium). In conclusion, the writer wishes to express his indebtedness to Prof. William H. Weston, Jr. for the loan of the preserved material of Clathrus columnatus Bosc. BIBLIOGRAPHY Fischer, E. (’86). Versuch einer systematischen Uebersicht деў die bisher bekannten Phalloideen. Kónigl. Bot. Gart. u. Mus. Berlin, Jahrb. 4: 1-92. pl.1. 1886. Lloyd, С. С. (09). Synopsis of the known Phalloids. рр. 48-65. f. 59-64. Michelius, P. A. (1729). Nova plantarum genera iuxta 'Tournefortii methodum dis- posita. p. 214. pl. 93. Florence, 1729 Rafinesque, C. S. (1808). Prospectus of Mr. Rafinesque Schmaltz's two intended works on North American botany; the first on the new genera and species of plants discovered by himself and the second on the natural history of the nguses, or mushroom-tribe of America. N. Y. Med. Repository, 2nd hexade 5 (11): 355. 1808. Saccardo, P. A. (788). Sylloge Fungorum 7: 18-21. 1888. Turpin, P. J. F. (1822). Dict. d. Sci. Nat. 25: 248. 1822. [Vor. 15, 1928] 112 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 20 Fig. 1. Laternea triscapa, showing the suspended, specialized receptacle bearing the gleba, and the smooth surfaces of the columns. Photograph of freshly collected specimen. Natural size. Fig. 2. Laternea triscapa. Receptacle and upper part of columns enlarged three times to show the definite development of tissue to form the receptacle. Fig. 8. Clathrus columnatus, showing the relatively smooth outer surface and the rough, pitted inner surface. The gleba may be seen still adhering to the, inner surface at the junction of the columns or receptacles. From preserved material collected in Porto Rico by P. V. Siggers. Natural size. Fig. 4-6. Clathrus columnatus. Fig. 4 shows the upper portion of the recep- tacle enlarged three times. There is no evidence of any tissue that may be con- sidered comparable to that found in the receptacle of L. triscapa, the gleba being applied to the inner pitted surface. Figures 5 and 6 show the receptacles spread out and viewed from below; the latter figure illustrates the division of one of the columns to form а 5-columnar receptacle. 4/5 natural size. ANN. Mo. Вот. Garp., Vor. 15, 1928 PLATE 20 LINDER—THE GENUS LATERNEA Аппајв of the Missouri Botanical Garden Vol. 15 APRIL, 1928 No. 2 SOURCES OF ENERGY FOR AZOTOBACTER, WITH SPECIAL REFERENCE TO FATTY ACIDS! Р, 1. GAINEY Soil Biologist, Kansas Agricultural Experiment Station Formerly Teaching Fellow in the Henry Shaw School of Botany of Washington University INTRODUCTION The thermo-chemical phenomena involved in the fixation of free nitrogen by various micro-organisms are not well under- stood. It has been assumed that the fixation process is endo- thermic in nature and that the necessary energy is, in the case of the Azotobacter group of organisms, derived from the oxidation of organic compounds, principally of a carbohydrate, acid, or alcohol nature. Regardless of whether the initial process through which nitrogen is brought into combination is exo- or endo-thermic, no one has been able to establish definitely a measurable fixation of nitrogen by Azotobacter, or any other nitrogen-fixing group of organisms, in the complete absence of some form of organic matter. Furthermore, growth and nitrogen fixation have been found to run more or less parallel with the quantity and nature of the organic material available, provided the material is non-nitrogenous in nature. It may be assumed safely, therefore, that an organic food material of some kind is essential in the metabolism of this group of organisms. This being true, it would seem highly desirable, both from a theoretical and practical standpoint, to secure as much information as possible relative 1 An investigation carried out in part at the Missouri Botanieal Garden in the Graduate Laboratory of the Henry Shaw School of Botany of Washington University and in part in the Research Laboratory of Soil Biology of the Kansas Agricultural Experiment Station, and submitted as a thesis in partial fulfilment of the require- ments for the degree of doctor of philosophy in the Henry Shaw School of Botany of Washington University. ANN. Мо. Bor. Garp., Vor. 15, 1928 (113) (Уот. 15 114 ANNALS OF THE MISSOURI BOTANICAL GARDEN to the different organic food substances suitable for these organ- isms and also the relative efficiency with which different com- pounds can be used. As an aid in the solution of some of the more complicated thermo-chemical questions involved it seemed desirable (0 ascertain, if possible, whether any quantitative relationship existed between the potential energy content of the organic material utilized, on the one hand, and the quantity of growth and nitrogen fixed, on the other. It was with the hope of securing information along these lines that this work was under- taken. More specifically this investigation has been concerned with seeking an answer to the following questions: (1) Is there any difference in the relative availability of the lower fatty acids as a source of carbon, or organic food substance, for Azotobacter? (2) If Azotobacter exhibits differences in ability to utilize different fatty acids, can such differences be associated with the structure, size, or energy content of the molecule? The following criteria were used in judging the ability of Azotobacter to utilize the various acids: (a) the variation in visible growth; (b) the disappearance of the acid; (c) the fixation of nitrogen; (d) changes in the hydrogen-ion concentration of the medium. The lower fatty acids were selected for study because they compose a series of compounds exhibiting many characteristics in common, yet in the series the molecule increases in definite increments from fairly simple to fairly complex. Furthermore, the heat of combustion of these compounds increases directly as the molecular weight increases. By including the iso com- pounds, two molecules varying in structure but identical in composition and energy content could be compared. An addi- tional desirable characteristic possessed by this series of com- pounds is that they are found as free acids or as constituents of fats in nature, and some of them are already known to serve as organic food for Azotobacter. One other characteristic essential in a series of compounds suitable for a study of this nature is susceptibility to fairly easy quantitative analysis. Not many series of organic compounds of which the members possess the 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 115 desirable characteristics enumerated above can be found, and possibly there is no other series in which as many members may be used so advantageously. Unfortunately, lack of solubility prevents any quantitative use of members of this series above six carbon atoms, but even then there would seem to be enough members of the series that can be used to give valuable informa- tion if carefully studied. METHODS General procedure—The general procedure has been to prepare, in one batch, as carefully as possible, all the medium neces- sary for a single experiment. Measured quantities of this were placed in the culture flasks which were then stoppered with cot- ton and sterilized in the autoclave. After sterilization the flasks were inoculated as uniformly as possible with a heavy suspen- sion of organisms washed from the surface of mannitol-soil-ex- tract agar upon which active vigorous growth was taking place. No old or apparently non-vigorous growing culture was used as an inoculum. After varying periods of incubation the cultures were removed from the incubator and the qualitative and quan- titative analyses were made as indicated. Where supplementary aeration was resorted to, an inlet tube of glass, containing five to seven small openings near the end, was inserted in a rubber stopper in such a way that when the stopper was tight in the culture flask the end of the tube almost reached the bottom of the flask. The stopper was also provided with an outlet tube to be connected to a vacuum system. The stopper and connecting tubes were sterilized separately and inserted after inoculation. Cotton was forced into the ends of the connecting tubes as a precaution against possible contamina- tion. In the aerated experiments 300-сс. Pyrex Erlenmeyer flasks were used as culture containers, and all those in any one experiment were connected in series so that the same quantity of aeration was provided for all. While incubating, a vigorous bubbling of air through the media was continuously maintained. Before entering the first culture flask the air was washed through flasks arranged as mentioned above, of acid, alkali, and water. Despite special precautions to prevent contamination there was one type of foreign organism difficult to keep out. [Vor. 15 116 ANNALS OF THE MISSOURI BOTANICAL GARDEN Where no special aeration was provided the cultures consisted of 50 се. of media in 300-сс. flasks; 100 се. media in 750-cc. flasks; or 200 се. media in 1000-сс. Pyrex Erlenmeyer flasks. These quantities of media in the flasks indicated always exhibited a large surface area compared to the depth, and while aeration was certainly not as vigorous as where air was drawn through the culture, nevertheless it was ample for very rapid growth. Growth at the bottom of the culture was frequently observed before it made its appearance on the surface, indicating aeration through- out the culture. These cultures were left stationary except when being handled for examination, and even then care was taken not to shake so vigorously as to break up any film that might be forming on the surface. Medium—Unless otherwise stated the medium used in the various experiments had the following composition, and its suitability is evidenced by the very rapid growth that took place when the organic material was assimilable: О ке Их, Л 2.50 gms. и Отт: ауы аа 20 g ә ДРИНА ЗУ оО РОТ 20 g АСЕ ОЕ M о сете 05 gm Foch (10 Por 7 СОРА 1 дгор Organic material..... ШОКТАШ ТҮҮЛ 1 per cent Е не ces 1000 се. In some of the preliminary experiments only 0.50 gm. of К,НРО, was used, but it was observed that when such a small quantity of phosphate was added the hydrogen-ion concentration sometimes changed so rapidly and markedly that growth was very soon inhibited by the increase of hydroxyl-ions. A rapid increase in hydroxyl-ion concentration was always observed when large quantities of a metallic salt of an organic acid were metab- olized, and probably arose from the formation of an hydroxide by the metallic-ions set free when the acid radicle was assimilated by the organisms. Even with 2.5 gms. phosphate and an excess of СаСО, the buffering effect was barely sufficient to permit of complete oxidation of 1.0 per cent acid. In fact, in some instances there is evidence to indicate that the high alkalinity accompanying vigorous oxidation not only prevented further activity, as is 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 117 evidenced by the failure of the organisms in certain cultures to oxidize all the acid even where abundant growth occurred, but actually resulted in the death of most or all of the organisms present. In case of some of the less soluble acids the quantity added was only 0.5 per cent. When the organic material was fatty acid it was, in all except a few preliminary experiments, added to the entire volume of distilled water. An excess of CaCO; was then added, and the material boiled until the reaction became neutral to brom-thymol- blue, indicating complete transformation into the calcium salt, after which it was filtered. This procedure was resorted to in order to hasten the completion of the reaction between the weakly dissociated acid and calcium carbonate. The other salts were then added, and if any change in the reaction took place it was again adjusted to neutrality by the addition of sodium hydroxide or sulphuric acid as needed. The medium was then measured into the culture flasks, care being taken to keep it well agitated in order to secure an equal distribution of the precipitate, a small quantity of СаСО; added, the flask plugged with cotton and sterilized in the autoclave at ten pounds pres- sure. The final reaction of medium prepared as indicated was never far from Ра 7.0. Obviously, one could not depend upon the original weight of any organic material subjected to the manipulations described in the preceding paragraph as indicating the final concentration. It was necessary, therefore, to prepare controls and make quanti- tative analyses of the final concentration of the organic materials in all cases. The agar used for maintaining stock cultures, for testing the purity of cultures, and for the preparation of the inoculum was a soil-extract-mannitol agar prepared as follows: One thousand gms. fertile garden soil were added to 1000 се. distilled water and subjected to fifteen pounds pressure in the autoclave for thirty minutes, after which СаСО; was added and the mixture filtered. The clear filtrate was made up to 1000 cc. with dis- tilled water. To 900 сс. distilled water was added 100 сс. soil extract, 0.5 gm. К.НРО,, 10 gms. mannitol, and 15 gms. agar agar. After heating in the autoclave to bring the agar agar into (Vox. 15 118 ANNALS OF THE MISSOURI BOTANICAL GARDEN solution, and while still hot, phenolphthalein and sufficient sodium hydroxide were added to give a distinct pink color. Cultures—The following include all cultures employed in any experiment, together with their origin. They were selected from among more than one hundred available cultures of Azoto- bacter. Those strains that were used to any appreciable extent were selected primarily because of their vigorous growing and nitrogen-fixing ability. Cultures Nos. За and 3b were strains of Azotobacter chroococcum secured from 5. А. Waksman, of New Brunswick, N. J. Cultures Nos. 4, 5a, 5b, 6, 7, 8, 57, 58, 59, 60, 62, and 66 were isolated from different Colorado soils in the laboratory of W. G. Sackett, Fort Collins, Colo. Culture No. 94 was a strain of Azotobacter vinelandii from the Bureau of Plant Industry, U. 5. Dept. Agr., Washington, О. С. Culture No. П was received from У’. Omeliansky, Leningrad, Russia. Culture No. 218, a strain of Azotobacter chroococcum marked “К,” was received from Chr. Barthel, Stockholm, Sweden. Cultures “С” and “В?” were received from the Rothamsted Experiment Station, England. “С” came originally from a single cell strain of H. В. Christensen’s and “В” was isolated from soil. Cultures Nos. 178, 187, 188, 194, and 165 were all isolated in this laboratory from the following soils, respectively: No. 178, Gloucester loam from Minnesota; No. 187, field soil from New York; No. 188, cotton and sugar cane soil, Virgin Islands; No. 194, soil from V. L. Winogradsky, Paris, France; No. 165, irrigated potato field soil from Wyoming. The only unidentified strain used to any appreciable extent was No. 62. This culture possessed the characteristics of Azotobacter chroococ- cum in that it grew abundantly as grayish-white opaque, distinct colonies, soon turning brown and eventually black with a more or less wrinkled dry surface. Before any culture was used in any experiment it was carefully tested for purity by repeated streaking and re-isolation from individual, microscopically examined colonies, until assured of the presence of only one type of organism. Furthermore, after incubation most cultures were again examined for purity and if evidence of contamination was present it has been so recorded. Inoculum.—The inoculum was prepared by streaking the 1928] GAINEY— SOURCES OF ENERGY FOR AZOTOBACTER 119 entire surface of a Kolle flask of soil-extract-mannitol agar with the desired culture, incubating 48-96 hours, or until the entire surface was covered with a uniform thick growth, and suspending this growth in 25-50 cc. of sterile water. This gave а suspension of such density as to be practically opaque in а depth of only half an inch. One or two per cent of this was used as the inoculum, thus insuring a very heavy inoculation. Incubation.—All cultures were incubated either at summer room temperature or in an incubator at 28-32° C. Room tem- perature was quite favorable in the summer but during the winter the temperature dropped too low at night for active growth. Most of the aerated experiments were run at room temperature, while all non-aerated experiments were incubated at 28-32° C. CHEMICAL METHODS Dextrose.—Quantitative dextrose determinations were made by the Shaffer-Hartmann (21) iodometric method. Where a heavy growth of Azotobacter had taken place the slime-like material present was precipitated by adding 1.0 per cent of a mixture of 2.5 gms. phosphotungstic acid and 5.0 gms. HSO, before sugar determinations were made. Preliminary experi- ments proved that sugar could be recovered quantitatively when added to a culture and treated by this method. Total nitrogen.—The Gunning modification of the Kjeldahl method was employed for total nitrogen. If the culture con- sisted of only 50 cc. of medium the entire volume was utilized, otherwise after making the culture up to the original volume 25-сс., or more frequently 50-cc., duplicated samples were run. А small piece of copper wire, 7 gms. of anhydrous sodium sulphate, and 35 cc. H.SO, were added and digestion continued for one hour after the solution became clear (see Latshaw, 16). Table 1 indicates the degree of accuracy with which duplicate determina- tions checked. In some of the experiments where aeration was employed the data for nitrogen determinations did not seem conclusive, and it has been thought best to leave them out entirely. No significance is attached to an increase in nitrogen of less than 0.5 mgs. per 100 cc. of medium, and all the data recorded in the tables are based upon 100 сс. [Vor. 15 120 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE I ACCURACY WITH WHICH TOTAL NITROGEN DETERMINATIONS CHECKED Mgs. nitrogen recovered from Sample No. ы Peptone solution | Peptone solution | Azotobacter culture 1 9.58 9.61 2.21 2 9.52 9.52 2.34 8 9.61 9.52 2.14 4 9.58 9.58 2.21 5 9.52 9.52 2.08 6 9.52 9.58 2.27 Average 9.55 9.55 2.25 Volatile acid.—Practically ай the acids used were Eastman Kodak Co. products. Quantitative determinations have been made by distilling 100 сс. from a total volume of 110 сс. Pyrex Erlenmeyer flasks of 300 cc. capacity connected to Liebig con- densers and surrounded by an asbestos shield were used as distillation flasks. These were heated by an electric hot plate, and it required 30-45 minutes, depending upon the acid, to dis- til over 100 сс. Titrations were made in increments of 20 сс. unless it had previously been noted that practically all acid had disappeared, in which case the entire 100 cc. were titrated at one time. This fractional titration was employed in order to enable the plotting of the titration curves to detect the trans- formation of a higher into a lower acid. Phenolphthalein was employed as an indicator, and care was exercised that all vessels and wash water were neutralized before being used. Figures 1 and 2 are given to show the relative titration curves of the different normal acids and also to show that there was no indication of an acid with a higher molecular weight being trans- formed into one of lower molecular weight. The curves for the standard acid solution and for the cultures in which abundant growth had taken place coincide as well as would curves from two different batches of acid. The culture distillation curves are from cultures in which approximately half the original acid had disappeared. Curves for iso compounds would show the same thing. These curves are plotted on a basis of the per cent of the total recovered that came over in each 20-cc. fraction, when 100 сс. were distilled from a total volume of 110 сс. 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 121 Calculations of the quantity of acid present were based upon quantitative distillations of carefully standardized acids distilled from pure water to which a small quantity of sulphuric acid had also been added. The data in table 11 show the per cent 50 ~ о = — -- ~ о ~ о Per cent of acid recovered — Standard so/ution Bx па ---Cu/tura/ solution 0 5 5 5 : : 3 е 3 м & ~ ~ ej = ш Fig. 1. Distillation curves for volatile acids from standard solutions and from cultures in which approximately half of the acid had been metabolized. of the total recovered from the different acids when varying quantities up to 100 сс. of the total 110 cc. had been distilled. Data recorded in table m1 show the degree of accuracy with which triplicate determinations checked. The figures in table [Vor. 15 122 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Iv indicate that the presence of the other constituents of the culture medium did not interfere with the recovery of the volatile acid. The quantity of medium distilled was, unless indicated to the contrary, 25 се., and duplicate or triplicate samples were always run. Only averages of the two or three checks are * © — Standard solution pense Cultural solution БЭ о Rr cent of acid recovered о / 20cc. 9 2% 20cc. F 20cc 4” 20cc Fig Distillation curves for volatile acids from standard solutions and from Ка in which approximately half of the acid had been metabolized. recorded. When distilled from the culture medium the volatile acid was freed from calcium by adding an excess of sulphuric acid. The СО, thereby liberated was removed by aerating vigorously for thirty minutes. In the aerated cultures, run for the longer periods of time, 1928] SOURCES OF ENERGY FOR AZOTOBACTER 123 GAINEY ралодооол ртов jo 1190 194 20:66 | 08:86 | 0286 100 007 | LF #6 109 #6 | OF 26 | OF #6 ЕР 82|09 82|06 22, | 08 82| 02:19 | 09 19 | 0619 | 09'19 | 001 07:96 | 06:96 | 06:96 | 06:96 | 2582, | 09' 82, | 0678 | 09-84 | 28:48 | OF 2€ | 09-6 08:26 | OZ TP | 01:19 OF TF| OT Tv | 08 28:16 | 02:16 | 06:16 (08:06 | ва 19 | 09-19 | 09:19 | 01:29 | OZ OF | OL OF | OL’ OF | OZ Or | 97-15 | OP 22 | 09' 22 | 0925 | 09 02:11 | 01'82 | 01'82 | 09:81 | 21:68 | 09-23 | 01:98 | 08:28 | 01:92 | 00:92 | 01:92 | 00:90 | 62:91 | 07:91 | 01721 |0/:91| 07 02:09 | 00:06 | 09:09 | 02:09 | от та | от те | от та | от та |22221 | 09'21 | 09'21 |09721 |018 |00 8 |088 |008 | 0 е с 6-1: е c 1 с с 4-1. е 2 Т әлү | зева | жем | лети | OAV | жез | sera | ев | ^^V | зевъ | asta | лева | ~ | же | лева | леша ша ІШІ отләү A огцот4о14 опооу огшлод 90 яауй па аллоо ватоу впотхул AO SNOLLV'TILLSIG ялуотатал HOIH.M HLIM хоучапооу AO яячона AHL ІП W'IGSV.L 0 001 0 001 Ру 66 1'96 Ӯ 46 6:86 6 16 Ӯ 76 8 08 с c9 001 0'001 0'001 8'16 с $6 8 76 9 66 с 06 6 18 9780 0 05 08 97646 0 001 2768 с 98 6°28 в 48 0 84 6 v9 $ IF с 92 09 9798 р 88 T 22 6 6L 8' IA 9'04 с 69 4 97 с 96 6 91 OF 8 19 T? 09 2704 а 97 6б 27 1 СР с СЕ с v6 98701 £4 05 отола во отола во it свој Зар“ зе аи 9LI9[VA от 3па от па | L К К | I 1 d -O8] [BULION ee Ч ы [BULION -081 puy | 971014054 қош зан рр рэлэлоээ1 1190 194 “ОО отт AO IVLOL У она G3'TILLSIG ячям ОО 001 кянм ватоу ЯТЫУТОл S001IHVA AO АЧЧАООЧЯ II ATAVL [Vor. 15 124 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE IV RECOVERY OF ACIDS FROM PURE WATER SOLUTION AND FROM AN AZOTOBACTER CULTURE MEDIUM; EXPRESSED IN EQUIVALENT QUANTITIES OF N/20 NaOH Butyric Valeric Саргоіс Propionic Ce. distilled H:O |Medium | НО {Medium | ЊО |Медит | НО |Medium 20 8.40 8.45 |11.40| 11.60 | 9.25| 9.45 | 5.95 5.95 40 14.70 | 14.95 |17.60| 17.90 |13.80| 14.00 |11.60| 11.95 60 19.50 | 19.80 |20.85| 20.95 |15.82 | 16.00 |16.60| 16.65 80 22.80 | 23.10 |22.10| 22.15 |16.55| 16.70 |21.10| 21.20 100 24.65 | 24.80 |22.56| 22.50 |16.75| 16.85 |25.00| 25.10 Total present |25.25| 25.45 |22.80| 23.00 |16.90| 17.05 |26 90| 9710 ег cent recovered 97.60| 97.50 |98.90| 98.00 |99.10| 98.80 |92.90| 92.60 there were indications in some instances that slight losses of acid occurred. This is not surprising, though, in view of the fact that despite four wash-bottles, designed to remove any acids and bases that might be present in the laboratory air as well as saturate the air with moisture, there were sometimes rather large losses in the volume of culture medium. Such losses were no doubt principally due to evaporation but probably in a less degree to the mechanical removal of moisture due to the bursting of so many bubbles. Owing to such discrepancies it has been difficult to evaluate accurately the utilization of acid by the growing cultures in certain instances. We have there- fore recorded as questionable such losses in the aerated experi- ments unless they obviously exceeded such losses where observa- tions were possible. Qualitative reaction.—It has been thought best to check roughly (time would not permit accurate determination) the hydrogen- ion concentration of the cultures in order to determine if the reaction were suitable for growth. For this purpose use has been made of brom-cresol-purple, brom-thymol-blue, phenol-red, cresol-red, phenolphthalein, and thymol-blue. Since the medium was only tested roughly as to whether it was acid, alkaline, or neutral to those indicators within whose range its reaction lay, the figures recorded in the various tables are merely approxi- mate. Only in those instances where the medium was found to be alkaline to thymol-blue and is recorded as 9.0 + has there been any indication that the reaction was unfavorable. In all 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 125 probability a medium alkaline to thymol-blue has an unfavorable, if not toxic, effect upon Azotobacter (see Johnson and Lipman, 722). PRELIMINARY EXPERIMENTS A large number of experiments of a preliminary nature have been performed. In fact, before any method, or step in a method, or culture was adopted for experimental use it was carefully tried to see that it would work satisfactorily. Among these preliminary experiments there are, aside from those already mentioned, a number that seem to be of sufficient significance to record. Experiments reported by Hunter (’23) indicated that by draw- ing a current of air through the medium, growth of Azotobacter and fixation of nitrogen could be greatly stimulated. Since Ше use of any method that would hasten growth seemed desirable, in view of the slow assimilation of certain of the fatty acids reported by Mockeridge (15), it was thought that Hunter’s method might be used advantageously. Therefore, an ехрегі- ment was designed not only to confirm Hunter’s results but at the same time to determine whether varying the rate of aeration would influence quantitatively the consumption of the organic food and the fixation of nitrogen. The results of such an experi- ment are reported in table v. No method of measuring quantitatively the rate of flow of air through the medium was available; however, in the samples subjected to “slow” aeration a slow continuous flow of bubbles, perhaps one a second, was maintained. In the “тейит” aerated cultures the air was drawn through at least ten times as rapidly, while Ше cultures subjected to “таріа” aeration probably received ten times as much air as the ‘‘medium”’ aerated cultures. The data show very definitely that increasing the rate of aeration increases both the rate of dextrose consumption and nitrogen fixation. There is some indication that the dextrose may possibly be utilized somewhat more efficiently with limited aeration, the average nitrogen-dextrose ratio being 1:79 for the “slow” aerated samples, whereas the corresponding ratio for the other samples was 1 : 115 and 1:113 respectively. [Vor. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN 126 еп РТУ 8 796 SII се Ч 209 61 861 201 э8в1элу SII ІР 8 796 611 сте СТР 06 $g I (241! 9 SII 10 8 796 101 58 9 069 OL 82 1 96 Y 911 10 8 796 921 28 v 909 14 28 1 801 $ еп 178 796 есі т9 9 569 28 те 1 801 6 801 688 796 9II 48 559 GL 861 TOI I pexg uos pexg peurnsuoo pexy ues pexg peurnsuoo рэху ues рэхц peurnsuoo -013ти "dur 194 | подолу овощхор | -oyu "Zur 194 | uoZoujtu әволхәр | -оддта “Эш Jed | uegozjru звощхер оволјхор “SBT ‘SZW за овощхер ‘SS "ЗИ "ЗИ оволјхор “Зүү вази Әр ње А „Ртазл,, uor voy „оплреш ,, uon u1oy ,,/AO[S,, попвло у SAVA ғ NOLLVAOONI “WE ОМ ячалттоо АЯ N390O?LLIN ЯО NOLLVXIJ ЯНІ аху ASOULXAG AO NOLLVZITILA AHL кода NOLLVUAV AO LOGAAH A W'IHVIL 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 127 In this connection it might be well to comment, in passing, upon the use of the term ''dextrose-nitrogen ratio." Certain investigators in speaking of this relationship have made use of the term ‘‘carbon-nitrogen ratio." This term, it is believed, does not accurately express the relationship. If it is a question of the organism securing or setting free a certain quantity of energy per unit of carbon, as is usually considered, obviously the expression C : № ratio is incorrect in that the energy freed per unit of carbon depends upon the other elements combined with the carbon as well as upon the carbon. For example, caproie acid contains approximately twice as much energy per gram of material and one and one-fourth times as much energy per gram of carbon as does dextrose, both being six carbon atom compounds. It would seem, therefore, that it would be much more logical to express this relationship upon an energy or molecular basis. In an effort to secure a desirable culture with which to carry out the more extensive investigations recorded in the next part of this paper, the experiments reported in tables уг and уп were designed. АП the cultures available at that time were included in these tests. TABLE VI VARIATION IN UTILIZATION OF DEXTROSE AND FIXATION OF NITROGEN BY DIFFERENT CULTURES OF AZOTOBACTER Ineubation 2 days Ineubation 5 days oe Mgs. Mgs. dextrose Mgs. Mgs. dextrose dextrose | nitrogen per mg. dextrose | nitrogen er mg. consume fixed nitrogen | consume fixed nitrogen fixed fixed 3a 169 2.20 77 620 4.96 125 3b 197 2.48 79 674 6.06 111 ‹ 171 1.38 124 522 1.10 474 а 179 2.06 82 679 3.58 189 5b 169 2.34 2 729 3.58 203 247 2.34 106 692 3.04 227 157 .82 192 250 .82 610 3 148 1.38 107 368 1.66 222 These data indicate that culture No. За was approximately as efficient in fixing nitrogen as any other. At the same time [У ог. 15 128 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE VII UTILIZATION OF DEXTROSE AND NORMAL BUTYRIC ACID* AND FIXATION OF NITROGEN BY DIFFERENT CULTURES OF AZOTOBACTER Бы қ ша extrose gs. utyric Culture a Mgs. used per | butyric Mgs. acid used No. pee М” тар. амд мутон ег mg чинээ - nitrogen | utilized кы nitrogen fixed fixed 3a 957 5.40 177 99 1.38 72 3b 957 4.55 210 98 1.76 55 4 168 3 00 -- ба 957 5.40 177 96 1.65 58 55 957 6.20 154 99 1,24 80 6 620 8.02 208 99 1.38 72 7 300 0 00 —- 8 957 8.44 281 99 82 121 * Butyric acid neutralized with sodium hydroxide. it grew abundantly, thus enabling ready detection of growth. It was also known to be a strain of Azotobacter chroococcum. For these reasons it was temporarily selected for further study. These data also indicate that certain cultures (Nos. 4 and 7), to all appearance Azotobacter, developed very poorly in the dextrose and not at all in the butyric acid medium. When measured by the quantity of nitrogen fixed per unit of organic material used, the actively growing cultures were apparently capable of utilizing butyric acid much more effectively than dextrose. In this particular experiment the increased effective- ness with which butyric acid was used might have been due to the relatively low per cent present, compared with dextrose. It has been frequently observed that in the presence of small quantities of organic material a higher fixation of nitrogen takes place рег unit of organic material consumed. Іп this experiment only 0.1 per cent butyric acid was present. Having selected culture No. За the next step was to test it in a preliminary way with several acids. Accordingly, the experiment recorded in table үш was arranged. For some unknown reason this culture was only able to utilize acetic acid among those tested. Similar results were secured in other experiments. Other cultures having been added to our collection, further qualitative tests were carried out, among which was the protocol 1928] GAINEY— SOURCES OF ENERGY FOR AZOTOBACTER 129 TABLE VIII UTILIZATION OF VARIOUS ACIDS BY CULTURE NO. 3A Mgs. recovered per 100 се. culture solution Acid Controls Incubation period in days Unaerated| Aerated 2 5 9 13 Formic 696 716 659 635 673 Acetic 1026 977 982 431 394 264 Ргорюше 1051 998 996 1002 Butyric 1018 1001 994 984 1001 Valeric 920 896 896 884 875 884 Dextrose 934 954 690 423 000 000 arranged in table rx. This experiment was also designed to gain some information relative to the effect of the cation as well as the acid radicle. The salts included were the only ones available of those particular acids at that time. The results presented in table rx indicate very strongly that the cation is probably of as much significance in determining the availability of an acid as is the anion. None of the formates permitted growth. This has been characteristic of formic acid in all tests conducted with it and is probably due to the formation of formaldehyde. Aluminum acetate appears to be the most readily available salt of acetic acid tested, all the strains being able to assimilate it readily. Uranium acetate, on the other hand, was not assimilated by any of the cultures. Between aluminum and uranium lay calcium, ammonium, magnesium, potassium, and sodium salts, their availability being approximately in the order given. mmonium acetate, while apparently assimilated by all ten cultures, supported vigorous growth only in two instances. It is possible that when supplied with nitrogen the very small quantities of organic impurities finding their way into the cultures enabled the various cultures to make perceptible growth. This, though, does not seem probable. Only half the cultures were capable of making visible growth when the sodium salt was the only source of organic matter supplied, and only one out of the ten assimilated it readily. Culture No. 60 could not even metab- olize dextrose readily. This experiment, as well as others reported in this paper, (Мог. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN 130 ($3ueuruedxo уподоутр тп цумола ојавлафшоо 859[ IO әлош эзвотрат suZrs 814 jo Joquinu oures әді) 'зпоштодхо re[norj1ed zey} ut родловао Y}MOIZ олтвјол эц) ојвотрит вийтв sn[d jo ледшти оца əya мола ојавиотвопђ војвотрш вш uonsenb в {434018 оп војвотрит ив впигш в војав) Эптрәәоопв в рив 5143 up, T s o “7 4 ер | + + Me v = 99 TE => T4. + ЖЭ a pete әр међ хан ме 59 ка Ээ” = é é é л л 4 деп x жүз 09 ғғ” Lum + - T Per 1 FET + "€" > шак 68 + ~ — 4 4 T pepe + = = = 388 Ф кені ti р тарын суми ИЙ EE Ре зе дек = Lg гет е5 + ЭЖ жет FEFA ын p T = "m зе 9 T4 мөч + + мес жк ДЕРТ ЭЭР нэ мэ Se ва ++ + — ... - 4 4d + +4 + ae Зөн == $ +++ ~ + F + «SR Бс ун тіл T кй e i ве 9384208 | 9484998 | 3787998 эүв)э28 | 9384208 | 918190% 9383998 | ојешлој | э ло} | эувшло) ‘ON 9801359 | цимива а) | urnrpog | штиввејод | шивэц рг unnep |зимиширу |umnruourury | umrpog |штојво | umruourury| по SAYA 8» GHLVHNONI "пятоуяотогту AO S4HALIOQO ъхячяляята АЯ ватоу OLLHOV аху опичоя AO SL'IVS БПОІЧУА AO NOLLVZI'ILLO ХІ W'IdV.L 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 131 indicates a very wide variability in the metabolism of organisms belonging to the genus Azotobacter. The data also emphasize the great need for more specific physiological studies of this very interesting group of organisms. It would appear utterly futile to attempt to apply the findings from the study of one strain or species to any other strain or species. Just ав Lóhnis and Smith (’16) have pointed out the futility of attempting to apply the morphological findings in any particular medium or at any particular time to the group as a whole, it is well to emphasize the same with regard to physiological studies. Since culture No. 62, apparently a strain of Azotobacter chroococ- cum and hence very closely related to culture No. 3a, seemed from the above-reported experiment, as well as from a number of unrecorded tests, to possess the ability to utilize a wider variety of salts of fatty acids than any other available culture, it was selected for the more intense studies reported in the next part of this paper. Also, even though aluminum acetate was undoubt- edly more readily available to some strains of Azotobacter than the calcium salt, the latter served equally as well for culture No. 62; and since calcium salts have found a much wider use in biological studies than aluminum it seemed desirable to use the calcium salt, thus making any results that might be secured more comparable with those reported by others. In addition, calcium salts are somewhat more easily prepared than aluminum. Calcium was therefore used as the basic element in succeeding studies. The question of the influence of the cation should certainly receive more study, and it is hoped that such studies may be continued in the near future. The data presented in table rx are only indicative of what may be expected. EXPERIMENTS WITH CULTURE No. 62 As previously mentioned, culture No. 62 probably belonged to the species Azotobacter chroococcum. Preliminary experiments indicated that it was a vigorously growing and strong nitrogen- fixing strain when supplied with a suitable form of organic material such as dextrose and certain of the lower fatty acids. The experiments conducted with this culture were all aerated. [Vor. 15 132 ANNALS OF THE MISSOURI BOTANICAL GARDEN Aeration was employed because the work of Mockeridge ('15) indicated that the rate at which certain organic materials were assimilated was extremely slow, long periods of incubation being necessary to insure appreciable utilization. Previous work had shown that the rate of growth, consumption of certain sugars, and fixation of nitrogen could be materially facilitated by draw- ing a current of air through the medium. It was hoped, by employing a similar method in these experiments, to shorten the time of incubation necessary to secure quantitative results of a definite character. Increased aeration unquestionably stimulated growth in many instances, but occasionally some difficulty was experienced in obtaining entirely satisfactory checking in quantitative nitrogen and volatile acid determina- tions following prolonged aeration, and in addition it was more difficult to maintain pure cultures. For these reasons the quan- titative experiments in which Azotobacter vinelandii was em- ployed were not aerated. Utilization of formic acid.—Experiments were carried out in which formic acid was used as the sole organic constituent of the medium, but there was no indication of either growth, utilization of the acid, or fixation of nitrogen, and therefore the data are not recorded. Utilization of acetic acid.—The data with regard to the utiliza- tion of acetic acid, recorded in table x, are quite conclusive in showing that the calcium salt of this acid is readily available to culture No. 62. Within seven days practically all the original 1.0 per cent of acid had disappeared, accompanied by abundant growth and a marked change in the reaction of the medium. In fact, it is probable that the hydroxyl-ion concentration was such as to inhibit further growth. Quite marked fixation of nitrogen was evident, but owing to an error in the method employed in the total nitrogen determinations in this experi- ment, the data are not recorded. Utilization of propionic acid.—The ability of culture No. 62 to utilize readily the calcium salt of propionic acid is quite evident from the data presented in table хі. Within nine days practically all the acid had disappeared, and a marked increase in the hydroxyl-ion concentration and nitrogen content of the cultures had occurred. 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 133 TABLE Х UTILIZATION OF ACETIC ACID BY CULTURE NO 62 Approximate M M M Flask | Days Purity reaction dd d шатн No. jneubated „т recovered | utilized fixed Control 2,8 1 0 Sterile 7.0-7.4 1059 —— 35 ы, Control "64 2 0 Sterile 7.0-7. 1062 ма 5 3 2 ure 7.0-7.4 31 229 984 4 2 Риге 7.0-7.4 906 154 "as 5 4 (a)* 7.0-7.4 668 392 яна 6 4 (а) 7.0-7.4 654 406 | 398 4 7 (а) 8.6-9.0 3 1024 Up 8 7 (a) 8.6-9.0 85 975 Чар 9 9 Риге 9. 0+ 39 1021 ны? 10 16 Contaminated| 9.0-- 48 1012 Sz = 11 16 ure 9.0+ 48 1012 858 12 22, Contaminated| 9.0+ 124 936 mee 13 22 Pure 9.0+ 91 969 a * Not tested. TABLE XI UTILIZATION OF PROPIONIC ACID BY CULTURE NO. 62 |Approximate Mgs. | Mgs. |Mgs. acid Flask uu Purit reaction ын acid | шїго- | used рег №. Би у expressed =. а utili- | gen | mg. nitro- Р assis В fixed | gen fixed Control 1 Sterile 7.0-7.4 958 — | — --- Contro 2 Sterile 7.0-7.4 952 — | — --- Control 3 Contaminated| 8.2-8.6 17 — | — — Contro 4 18 Sterile )-7.4 974 5 3 7.0-7.4 829 132 76 6 5 Pure 7.0-7.4 705 256 | 4.74 54 7 7 Pure 7.0-7.4 626 335 92 8 9 9 21 940 — 9 12 Риге 9.0+ 37 924 ; 196 10 18 Contaminated} 9.0+ 17 944 — — In this experiment control flasks Nos. 1 and 2 were not aerated, while Nos. 3 and 4 were aerated under the same conditions and for as long a time as any of the inoculated flasks. The culture adjacent to control culture No. 3 foamed badly, resulting in the contamination of No. 3 with Azotobacter; therefore it is not considered in the quantitative calculations. It is evident, [Vor. 15 134 ANNALS OF THE MISSOURI BOTANICAL GARDEN though, from a comparison of control flasks No. 1 and No. 2 with No. 4 that no volatilization of the propionic acid took place. Therefore the decrease in the quantity of propionic acid that occurred in the presence of pure cultures must have been due to its assimilation by the organisms. Utilization of normal butyric acid.—The data presented in table хи show that culture No. 62 is also capable of utilizing normal butyric acid in its metabolism. Again only seven days were required for almost complete assimilation of the acid present, with corresponding decreases in the hydrogen-ion con- centration. The quantities of nitrogen fixed were also marked, as was the visible growth of the organisms. TABLE XII UTILIZATION OF NORMAL BUTYRIC ACID AND FIXATION OF NITROGEN BY CULTURE NO. 62 Approximate | Mgs. | Mgs. | Mgs. acid Flask Days Purit reaction M acid | nitro-| used per No. еме нт expressed мэ d utili- | ge mg. nitro- мэ. аз Рн есейдім zed | fixed | gen fixed Control 1 Sterile 7.0-7.4 978 — | — — Control 2 0 Sterile 7.0-7.4 995 — 3 2 7.0-7.4 956 30 .0 334 4 2 Риге 7.0-7.4 878 108 | 1.25 86 5 5 Риге Т.0-7.4 706 280 | 2.73 102 6 5 Риге 7.0-7.4 669 17 | 4.15 76 7 7 Риге 7.0-7.4 439 47 | 2.59 212 8 12 Риге 9.0+ 78 908 | 6.77 184 9 17 Contaminated| 9.0+ 61 925 | 4.95 186 10 17 Contaminated] 9.0+ Lil 875 | 6.29 140 Utilization of iso-butyric acid.—Under the experimental condi- tions to which the cultures recorded in table хит were subjected there was little or no indication that culture No. 62 could utilize iso-butyric acid in its metabolism. 16 is true that there was some decrease in the quantity of volatile acid present in the different cultures but the quantities were small. Besides, the non-inocu- lated sterile aerated controls, No. 2 and No. 3, showed practically the same decrease as inoculated flask No. 10, incubated the same length of time. In addition there was no perceptible change in the reaction, and the quantities of nitrogen fixed, if any, did not exceed the experimental error. Visible growth was also 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 135 questionable. It seems safe, therefore, to conclude that culture No. 62 could not utilize the calcium salt of iso-butyric acid under the conditions obtaining in these experiments. TABLE XIII UTILIZATION OF ISO-BUTYRIC ACID BY CULTURE NO. 62 Approximate Flask Days Purit reaction |Mgs. acid|Mgs. acid|Mgs. nitro- No. |incubated чат expressed |recovered| utilized | gen fixed as Py Control 1 0 Sterile 7.0-7.4 831 None None Control 2 Sterile 7.0-7.4 774 None None Control 3 17 Sterile 7.0-7.4 773 Мопе None 4 2 ure 7.0-7.4 886 None None 5 2 Pure 7.0-7.4 800 None None 6 5 Contaminated) 7.0-7.4 779 None None Ч 5 Ра 7.0-7.4 778 None None 8 4 Contaminated 7.0-7.4 799 None None 9 12 Pure 7.0-7.4 746 None None 10 Tw. C 7.0-7.4 758 None None Utilization of normal valeric acid.—Not only was normal valeric acid not available to culture No. 62 but it actually was sufficiently toxic to kill all the introduced organisms within three days. ‘This is the only instance in which any acid studied, other than formie, has actually killed the culture except when marked change in reaction occurred. There was of course по utilization of the acid or fixation of nitrogen. The slight loss of volatile acid previously referred to is evident in the data presented in table хту. Utilization of monohydrated valeric acid.—The data presented in table xv with regard to this acid are inconclusive. There is a distinct loss of acid, evidently not due to its removal through aeration, because uninoculated aeration controls No. 3 and No. 4 incubated one day longer than the longest incubated inoculated flask showed no loss in volatile acid. On the other hand, there was no perceptible change in reaction, and the quantities of nitrogen fixed, if any, were too small to detect, there being no ! Mono- and trihydrated valerie acids were iso compounds made by Merck and Co. [Vor. 15 136 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE XIV UTILIZATION OF NORMAL VALERIC ACID BY CULTURE NO. 62 Approximate Flask Days Purit reaction в. acid|Mgs. acid|Mgs. nitro- No. |jincubated — ec recovered | utilized | gen fixed as Control 1 0 Sterile 6.6-7.0 992 None None Control 2 0 Sterile 6.6–7.0 992 Мопе Копе 3 3 erile 6.6–7.0 988 Мопе Копе 4 6 Contaminated| 6.6-7.0 890 Мопе Копе 5 11 Sterile 6.6-7.0 990 None None 6 11 Sterile 6.6-7.0 913 None None 7 18 Sterile 6.6–7.0 973 Мопе None 8 18 Sterile 6.6-7.0 979 Мопе Мопе 9 32 бїегїїе 6.6-7.0 935 None None 10 32 Sterile 6.6-7.0 951 Мопе Мопе TABLE ХУ UTILIZATION OF MONOHYDRATED VALERIC ACID BY CULTURE NO. 62 Approximate Flask Days Purit i Mgs. ас Мрз. acid|Mgs. nitro- No. | incubated — expressed |recovered| utilized | gen fixed as Рн Control 1 Sterile 7.0-7.4 1055 -- None Control 2 0 Sterile 7.0-7.4 1058 ---- Мопе Control 3 33 Sterile 7.0-7.4 1064 --- Мопе Control 4 33 Sterile 7.0-7.4 1066 — Мопе 5 8 ге .0-7.4 1051 10 None 6 6 ure 7.0-7.4 944 ІМ None 7 11 Риге 7.0-7.4 911 150 None 8 11 Риге 7.0–7.4 972 89 Мопе 9 18 Риге 7.0-7.4 899 162 Мопе 10 18 Риге 7.0-7.4 907 154 Мопе 11 82 Риге 7.0-7.4 871 190 Мопе 12 82 Риге 7.0-7.4 933 128 None difference in the nitrogen content of flasks No. and sterile controls No. 3 and No. 4 Utilization of trihydrated valeric acid.—Here again the quanti- ties of volatile acid not recovered and increases in total nitrogen were not sufficient to be regarded as significant. of acid disappearing are based upon aerated control No. 3 no losses are evident. 11 and No. 12 If the quantities Unfortunately, this sample became con- 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 137 taminated, and conclusions based upon it would not be entirely valid. On the other hand, if the quantities of acid recovered from the various flasks are compared with those recovered from controls No. 1 and No. 2, analyzed at the beginning of the experi- ment, small losses of acid are evident. What has been said with regard to losses of acid is equally true of increases in nitrogen. It is preferred, therefore, to regard it merely as a questionable possibility that culture No. 62 assimilates this acid qualitatively. The quantitative utilization of this acid, as well as of the mono- hydrated sample, is certainly small, if it occurs at all, compared to that of some of the other acid studied. TABLE XVI UTILIZATION OF TRIHYDRATED VALERIC ACID BY CULTURE NO. 62 Approximate Flask Days Purit reaction |Mgs. acid|Mgs. acid|Mgs. nitro- No. | incubated пра expressed |recovered| utilized | gen fixed as Py Control 1 0 Sterile 7.0-7.4 784 2 е Control 4 2 2 Sterile 7.0-7.4 785 Е E Control B 2 3 19 Contaminated} 7.0-7.4 726 Ф E: 4 3 Pure 7.0-7.4 719 5 Е 5 3 Pure 7.0-7.4 776 д "i 6 7 Pure 7.0-7.4 756 3 8 7 7 Contaminated| 7.0-7.4 752 a d 8 13 Pure 7.0-7.4 766 3 8 9 18 Риге 7.0-7.4 782 > Ёс 10 19 Contaminated| 7.0-7.4 748 11 19 Contaminated 7.0-7.4 756 Utilization of normal caproic acid.—The data presented in table хуп show beyond question that the calcium salt of normal caproic acid may serve as a readily available source of organic material for culture No. 62. The samples analyzed after seven days’ incubation, while still containing large quantities of volatile acid, showed marked changes in reaction, appreciable losses of acid, and fixation of nitrogen. The samples analyzed after thirteen days still contained appreciable quantities of acid but the hydroxyl-ion concentration had probably reached a point where growth was inhibited. This is further indicated by the failure to detect further losses of acid in the flasks incubated for a longer time, No. 10 and No. 11. 138 TABLE XVII ANNALS OF THE MISSOURI BOTANICAL GARDEN (Уот. 15 UTILIZATION OF NORMAL CAPROIC ACID BY CULTURE NO. 62 | Approximate | Mgs. gs. |Mgs. acid Flask x a Purit 1 ар, acd nitro-| used per No. "ien T expressed ~ } sed utili- | gen | mg. nitro- - — В ed | gen fixed Control 1 0 Sterile 7.0-7.4 408 — | — — Control 2 19 Contaminated} 9.0+ 137 — | — -- Control 3 19 Contaminated} 7.0-7.4 359 — --- 4 8 Риге 6.6-7.0 351 57 | 1.02 56 5 8 Риге 7.0-7.4 874 84 . 98 34 6 T Pure 7.6-8.0 337 71 .50 142 7 7 Contaminated| 7.8-8.2 260 148 | 2.16 69 8 13 Contaminated| 9.0+ 87 321 | 2.96 104 9 13 Contaminated) 9.0+ 75 333 | Lost — 10 19 Риг 9.0-- 102 806 | 4.80 64 11 19 Риге 9.0+ 109 299 | —— -- Utilization ој iso-caproic acid.—While the data presented in table хупт may be regarded as inconclusive, they nevertheless indicate that iso-caproic acid is not available as an organic food for culture No. 62. There were slight losses of acid, but such losses were equally as marked from the non-inoculated, sterile, aerated controls as from any inoculated cultures. Further- TABLE XVIII UTILIZATION OF ISO-CAPROIC ACID BY CULTURE NO. 62 Approximate Flask Days Purit i Mgs. acid|Mgs. acid|Mgs. nitro- No. | incubated ид expressed | гесоуегей| utilized | gen fixed ав Рн Control 1 Sterile 7.0-7.4 560 Control © Ф 2 Sterile 7.0-7.4 583 4 5 Control 8 8 3 Sterile 7.0-7.4 520 B 3 Control 8 Ed 4 19 terile 7.0-7.4 497 = 5, 5 3 ure 7.0-7.4 526 а 6 3 Риге 7.0-7.4 --- ‚© 8 7 7 Риге 7.0-7.4 522 3 * 8 7 Pure 7.0-7.4 505 E E: 9 13 Pure 7.0-7.4 517 = <>) 10 13 Contaminated| 7.0-7.4 493 P 11 19 7.0-7.4 513 12 19 [Sterile 7.0-7.4 518 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 139 more, there was no appreciable changes in reaction or detectable increases in the nitrogen content. Summary of experiments with culture No. 62.—Under the experimental conditions to which culture No. 62 was subjected in the experiments herein reported the calcium salts of formic and normal valeric acids were strongly germicidal. Similar salts of acetic, propionic, normal butyric, and normal caproic acids served as readily available sources of organic food. The other salts tested, namely, iso-butyric, mono- and trihydrated valeric and iso-caproic, were either not available or utilized very slowly and in small quantities. EXPERIMENTS WITH AZOTOBACTER VINELANDII The culture of Azotobacter vinelandii used in these experiments (Culture No. 94) was obtained from N. R. Smith, of the Bureau of Plant Industry, U. $. Department of Agriculture. It grew vigorously upon soil-extract-mannitol agar, less vigorously upon beef-extract agar, and produced the typical green fluorescence upon the former medium. It also grew very abundantly in the liquid medium employed when dextrose, mannitol, and calcium salts of several of the fatty acids were supplied as the organic material. Utilization of formic acid.—There was no evidence either in the qualitative or quantitative experiments of the ability of this organism to utilize calcium formate and therefore the quantita- tive data are omitted. Utilization of acetic acid.—An examination of the data рге- sented in table хіх should convince any one of the ability of this culture to utilize readily the calcium salt of acetic acid. A very marked change in the reaction, an almost complete disappearance of the acid accompanied by definite fixation of nitrogen, together with an abundance of visible growth, sub- stantiate the above conclusions. In this experiment the hydroxyl- ion concentration evidently reached a point that could not be tolerated by the organisms, most of them being dead when the flasks incubated for the longer period of time were examined. Utilization of propionic acid.—Azotobacter vinelandii can readily assimilate propionic acid under the conditions obtaining in the 140 TABLE XI X ANNALS OF THE MISSOURI BOTANICAL GARDEN [Vor. 15 UTILIZATION OF ACETIC ACID AND FIXATION OF NITROGEN BY AZOTO- BACTER VINELANDII Approximate} gs. | Mgs. acid Flask Part Рие reaction ү >] add nitro-| used per No. bate ^ expressed омања utili- en | mg. nitro- as Py — ed | fixed | gen fixed 1 |Control | Sterile 7.0-7.4 876 — | — --- 2 | Control | Sterile 7.0-7.4 859 — | — — 8 4 ur 7.0-7.4 783 84 | —.06 — 4 4 Pure 7.0-7.4 790 T4 .20 384 5 9 ure 7.0-7.4 667 200 | —.20 --- 6 9 Риге 7.0-7.4 575 292 .12 440 7 14 Риге 7.0-7.4 428 444 ‚50 888 8 14 ure 7.0-7.4 389 478 .96 498 9 24 Риге* 9.0 8 864 1.60 10 24 ure* 9.0+ 3 864 3.06 282 11 33 Sterile 9.0+ 5 862 | 2.36 366 12 33 ure* 9.0+ 11 856 2.22 386 * Very few living organisms. experiment reported in table xx. Within three weeks the original 1.0 per cent of acid had practically disappeared. The abundance of visible growth, the marked increase in Будгоху!- ion concentration, and definite increases in the nitrogen content of the cultures are additional proof of the ability of the organism to assimilate this particular acid. TABLE XX UTILIZATION OF PROPIONIC ACID AND FIXATION OF NITROGEN BY AZOTO- BACTER VINELANDII | Approximate Мрз. | Mgs. |Mgs. acid Flask mun Podés reaction => мал nitro-| used рег о. bat d иу expressed neared utili- | gen | mg. nitro- as Py 2 fixed | gen fixed 1 | Control | Sterile 7.0-7.4 1009 — | — --- 2 | Control | Sterile 7.0-7.4 999 — | — — 3 7 ur 7.0-7.4 790 214 3 668 4 14 Pure 7.0-7.4 575 429 | 1.46 194 5 28 Риге ).О-+ 16 88 | 5.92 168 6 28 Риге ).04- 45 959 | 6.06 158 T 30 ure ).04- 7 997 | 4.46 224 8 30 Sterile ).04- 10 994 | 4.02 247 9 39 Sterile ).04- 7 997 | 3.12 319 10 39 Sterile 0-- Ч 997 | 2.86 422 11 46 Sterile 0-- 4 1000 | 2.22 450 12 46 Sterile 0-- 6 998 | 3.12 319 Utilization of normal butyric acid.—Some irregularities were exhibited in the growth in different culture flasks containing 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 141 normal butyric acid and inoculated with Azotobacter vinelandii despite all efforts to make the duplicate flasks as nearly identical as possible. These irregularities are reflected in the quantity of acid unrecoverable, the changes in reaction, and in the increases in nitrogen content as recorded in table ххі. However, these data unquestionably show a ready utilization of this acid by the culture in question. The rapidity of disappearance of the acid is not as great as with acetic and propionic acids, but the increases in nitrogen per unit of acid assimilated are greater. TABLE ХХІ UTILIZATION OF NORMAL BUTYRIC ACID AND FIXATION OF NITROGEN BY AZOTOBACTER VINELANDII Approximate} Мрз. | Мез. | Mgs. acid Flask Days Purit reaction Mg Ч acid | пиго- | used рег Мо. зарн ШОУ? expressed энэ а | utili- | gen | mg. nitro- as Рн —— zed | fixed | gen fixed 1 |Control | Sterile 7.0-7.4 806 — | — --- 2 | Control | Sterile 7.0-7.4 819 — | —— --- 3 9 Риге 7.0-7.4 761 51 „12 424 4 9 Риге 7.0-7.4 750 2 .38 162 5 16 Риге 7.0-7.4 00 112 .76 148 6 16 Риге 7.0-7.4 00 112 . 64 176 7 25 Риге 7.0-7.4 499 313 | 2.62 118 8 25 Риг 7.0-7.4 314 498 | 5.80 9 9 39 Sterile 9.0+ 46 766 | 6.12 126 10 39 Pure 8.4-8.8 321 491 | 3.82 128 1 53 Pure 9.0+ 51 761 | 6.12 124 12 53 Pure 8.8-9.2 411 401 | 3.56 112 Utilization of iso-butyric acid.—The data presented in table ххп indicate rather strongly that the calcium salt of iso-butyric acid is not nearly so readily available as is the corresponding salt of the normal acid. Even after forty-nine days over half of the original acid was still present, the reaction had changed only slightly, and the quantity of nitrogen fixed was small compared with that fixed where the normal acid was present. Besides, the visible growth (unmistakably present) was also small compared to that where acetic, propionic, or normal butyric acid was the source of organic food. Utilization of normal valeric acid.—This acid apparently was not assimilated by Azotobacter vinelandii as readily as some of the lower members of the series. However, the figures presented in table ххіп show practically complete disappearance in flasks [Vor. 15 142 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE XXII UTILIZATION OF ISO-BUTYRIC ACID AND FIXATION OF NITROGEN BY AZOTOBACTER VINELANDII Approximate Mgs. | Mgs. |Mgs. acid Flask Days Purit reaction => edd nitro-| used per No atad жит expressed |, №. red| Utili- | gen | mg. nitro- шэн аз Рн OI m fixed | gen fixed 1 | Control | Sterile 7.0-7.4 835 — |--- — 2 | Control | Sterile 7.0-7.4 846 -- 3 T ure 7.0-7.4 795 45 | Lost --- 4 7 Риге 7.0-7.4 790 50 | Говђ шин 5 14 Риге 7.0-7.4 753 87 .26 334 6 14 Раге 70-7.4 Lost —— 32 c 7 17 ure 7.0-7.4 700 140 12 шин 8 17 Риге 7.0-7.4 728 112 174 9 26 Риге 7.0-7.4 662 178 278 10 26 Риге 7.0-7.4 634 206 322 11 38 Риге 7.0-7.4 495 343 | 1.22 282 12 49 Риге 7.0-7.4 467 373 | 1.14 338 Nos. 9, 11, and 12. The more rapid assimilation in these in- stances, though, might have been partially due to the contamina- ting organisms. That there was, however, unmistakable utiliza- tion in the uncontaminated flasks, Nos. 7, 8, and 10, is proved by the decreased acid content, change in reaction, and increase in nitrogen content. Both the total quantity of nitrogen fixed and the relative quantity fixed per unit of acid assimilated were large, the latter being greater than for any lower member of the fatty acid series. TABLE XXIII UTILIZATION OF NORMAL VALERIC ACID AND FIXATION OF NITROGEN BY AZOTOBACTER VINELANDII | Approximate Mgs. | Mgs. | Mgs. acid Flask и ж Purit reaction р =} acid | nitro-| used рег о. Dated у expressed мори utili- | gen | mg. nitro- ze fixed | gen fixed 1 | Control | Sterile 7.0-7.4 892 — | — --- 2 | Control | Sterile 7.0-7.4 883 8 7 ure 7.0-7.4 810 77 ‚70 110 4 7 u 7.0-7.4 835 52 .70 5 14 Contaminated| 7.0-7.4 770 117 .96 122 6 14 Contaminated| 7.0-7.4 750 137 | 1.34 102 7 26 8.8-9.2 197 90 .68 102 8 26 Pure 8.8-9.2 621 266 | 2.80 9 38 Contaminated| 9.0+ 833 | 6.76 122 10 38 Pure 7.4-7.8 414 473 | 6.56 11 49 Contaminated] 9.0+ 825 |7. 108 12 49 Contaminated| 9.0+ 827 111.98 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 143 Utilization of monohydrated valeric acid.—Growth in the pres- ence of this acid was slow, and the total amount appeared to be much less than with the normal valeric or with the acids of smaller molecular weight. This was reflected in the rate and total disappearance of the acid as well as in the quantity of nitrogen fixed. It is also evident from the data recorded in table xxiv that slight, if апу, change took place in the hydrogen-ion con- centration. However, the amount of nitrogen fixed and acid consumed, coupled with the presence of visible growth, show conclusively that this acid can be assimilated by the culture in question, but probably not as readily as the other low molecular weight straight-chain, fatty acids, at least not under the condi- tions of these experiments. TABLE XXIV UTILIZATION OF MONOHYDRATED VALERIC ACID AND FIXATION OF NI- TROGEN BY AZOTOBACTER VINELANDII Approximate Mgs. | Mgs. |Mgs. acid Flask Days Purit reaction зэр, acid | nitro-| used рег No arr е ехргеввей Эл q| utili- | gen | mg. nitro- "— аз Рн ыа M^ fixed | gen fixed 1 | Control | Sterile 7.0–7.4 98 — | —— шин 2 | Control | Sterile 7.0-7.4 1021 3 9 Pure 7.0-7.4 83 192 64 300 4 9 Риге 7.0-7.4 830 175 .76 230 5 21 Риге 7.0-7.4 816 189 | 1.40 186 6 21 Риге 7.0-7.4 841 1.52 108 7 28 Риге 7.0-7.4 685 320 | 2.62 122 8 28 Риге 7.0-7.4 714 291 | 1.52 192 9 88 Риге 7.0-7.4 637 368 | 2.04 180 10 88 Риге 7.0-7.4 712 293 | 1.66 176 11 49 Риге 7.0-7.4 631 874 | 1.90 196 12 49 Риге 7.0-7.4 663 842 | 2.16 158 Utilization of trihydrated valeric acid.—What was said with regard to the monohydrated valeric acid also applies to the trihydrated, except that the quantities of acid assimilated and the quantities of nitrogen fixed were smaller. No perceptible change took place in the hydrogen-ion concentration, and only one-fifth of the acid was not recoverable after seven weeks of incubation. These facts would indicate that this acid is assimi- lable by Azotobacter vinelandii with somewhat more difficulty than either the normal or monohydrated valeric acids. The data are presented in table xxv. [Vor. 15 144 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE XXV UTILIZATION OF TRIHYDRATED VALERIC ACID AND FIXATION OF NI- TROGEN BY AZOTOBACTER VINELANDII | Approximate | Mgs. | Mgs. |Mgs. acid Flas Days Purit reaction Megs. acid nitro-| used рег No. Pa "У ехргеззе — d utili- | gen | mg. nitro- " аз Рн гесоуегейі zed | fixed | gen fixed 1 | Control | Sterile .0-7.4 784 — |-- -— 2 | Control | Sterile ' 0-7.4 788 — | — -- 8 7 Риге .0-7.4 780 56 38 148 4 7 Риге ".0-7.4 753 33 26 122 5 14 Риге ' 0-7.4 729 57 26 218 6 14 Contaminated ' 0-7.4 714 72 32 224 T 24 Pure ".0-7.4 752 34 50 68 8 24 Риге ".0-7.4 708 78 88 88 9 85 Риге ".0-7.4 656 180 | 1.28 116 10 85 Риге ".0-7.4 668 118 | 1.22 96 11 49 Contaminated| 0-7.4 632 154 76 202 1 49 Риге 7.0-7.4 628 158 64 246 Utilization of normal саргос acid.—It would appear from the information recorded in table ххут that normal caproic acid can be metabolized by Azotobacier vinelandii the most readily of any fatty acid tested. Within four days a very heavy growth was evident, half the acid added to the medium had disappeared, the reaction had become alkaline to thymol-blue, and marked fixation of nitrogen had taken place. Within nine days the volatile acid had reached the minimum recorded for any incuba- tion period. Furthermore, the quantity of nitrogen fixed per TABLE XXVI UTILIZATION OF NORMAL CAPROIC ACID AND FIXATION OF NITROGEN BY AZOTOBACTER VINELANDII | Approximate | Mgs. | Mgs. | Mgs. acid Flas P " Purit reaction Mgs add nitro-| used per No. b Е а у expressed = d utili- | gen | mg. nitro- 5 ав Рн хаслланй OF fixed | gen fixed 1 | Control | Sterile 7.0-7.4 408 — | — ман 2 | Control | Sterile 7.0-7.4 381 —— 3 + Риге 9.0+ 267 128 | 3.92 32 4 4 Риге 9.0+ 180 215 | 4.66 46 5 9 9.0+ 39 356 | 5.86 60 6 9 Риге 9.0+ 41 354 | 5.48 64 7 16 Риге 9.0+ 32 363 | 6.06 60 8 16 Риге 9.0+ 28 367 | 6.62 56 9 28 Contaminated| 9.0+ 27 368 | 5.28 70 10 28 Contaminated| 9.0-- 31 364 | 6.50 56 11 37 Contaminated| 9.0+ 35 360 | 8.34 82 12 37 9.0+ 35 360 | 5.10 70 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 145 unit of acid assimilated was greater than for any other acid tested quantitatively. This would seem to indicate that increas- ing the size of the molecule does not necessarily decrease its availability. Utilization of tso-caproic acid.—This acid was, according to the data presented in table ххуп, readily assimilated, though the rate of growth, acid utilization, and nitrogen fixation were not equal to the corresponding rates where normal caproic acid was the sole organic food supplied. Similarly, the ratio between acid consumed and nitrogen fixed was twice as wide as for the normal acid. The iso compound then, it would seem, is not only less readily metabolized but can also not be used as economically as the straight-chain molecule of this acid. TABLE XXVII UTILIZATION OF ISO-CAPROIC ACID AND FIXATION OF NITROGEN BY AZOTOBACTER VINELANDII Approximate} Mgs. | Mgs. | Mgs. acid Flask Days Ран reaction Mgs. id | пиго- | used рег incu- urity d acid ‘li О: Бара ехргеззе recovered utili en | mg. nitro- ав Z fixed | gen fixe 1 | Control | Sterile 7.0-7.4 417 — | — шин 2 | Control | Sterile 7.0-7.4 409 — |-- 8 4 те 7.88.2 323 90 .50 180 4 4 Риге 7.8-8.2 272 131 .90 144 5 16 Риге 3. 2-8 .6 9 323 | 2.28 142 6 16 Риге 3, 2-8.6 75 358 | 2.16 156 T 28 Pure ).04- 23 390 2.54 154 8 28 Pure ).04- 22 391 3.18 122 9 37 Pure ‚0+ 77 336 3.70 90 10 37 Pur .0-- 23 390 2.80 140 11 51 Sterile 9.0+ 17 396 54 156 1 51 Риге* 9.0+ 15 398 68 148 5 Only four colonies developed on two plates. Summary of experiments with Azotobacter vinelandii (culture No. 94).—Azotobacter vinelandii is capable of growing in a medium containing as the only organic material the calcium salt of the following fatty acids: acetic, propionic, normal butyric, iso- butyric, normal valeric, iso-valeric (monohydrated valeric and trihydrated valeric), normal caproie, and 180-саргоје. This organism failed to grow under similar conditions in the presence of calcium formate. The rate of growth varies with the different calcium salt added (Vou. 15 146 ANNALS OF THE MISSOURI BOTANICAL GARDEN to the medium, being very rapid with normal caproic and very slow with all of the iso acids tested. When growth takes place there is an increase in the nitrogen content of the medium and a decrease in the quantity of volatile acid. The quantity of nitrogen fixed in the presence of any given acid corresponds more or less closely with the quantity of acid disappearing. The increase in nitrogen per unit of acid con- sumed by the organisms varies with different acids. If only normal acids are considered the quantity of nitrogen fixed per unit of acid decomposed increases as the molecular weight of the acid increases. Growth, disappearance of acid, and increase in nitrogen are not as rapid where iso acids are added to the medium as when the acid is a normal compound. EXPERIMENTS WITH OTHER CULTURES Since there were a number of instances in which the results secured with cultures No. 62 and No. 94 did not agree, it seemed desirable to extend somewhat similar tests to other cultures. In order to secure very active nitrogen-fixing strains for these tests the experiment recorded in table ххупт was arranged. It was also hoped that this experiment would show to what extent vigorous growth in liquid media, utilization of dextrose, and fixation of nitrogen could be correlated. Erlenmeyer flasks of 300 cc. capacity containing 1.0 per cent dextrose medium were prepared and inoculated heavily with the cultures indicated. One of the triplicate flasks was used for qualitative sugar tests. Growth observations were recorded from the remaining two flasks and after fifteen days’ incubation the quantity of nitrogen fixed was determined. Since the dextrose had completely disappeared in all but two instances and the original quantities in the various cultures were identical, the total nitrogen figures represent approximately the quantity of nitrogen fixed per 1000 mgs. dextrose consumed. It will be noted that with few exceptions the quantities of nitro- gen fixed do not vary very widely. The smallest quantity fixed in any instance where complete disappearance of sugar had taken place was 2.64 mgs. while the largest was 11.46 mgs. Most of the cultures showed a fixation of about 8 to 10 mgs. per 1000 1928] PRESENCE OF, AND ТО ASSIMILATE RELATIVE ABILITY TO FI GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER TABLE XXVIII THE ABILITY OF VARIOUS AZOTOBACTER CULTURES TO GROW IN THE 147 DEXTROSE, AND THEIR X NITROGEN А Nitrogen Quantity of growth after Presence of dextrose бање агу! 1048 of after varying рег- -— җы "I AIO incubation iods of incubation == им 2 даув 4 даув 8 days 8 даув 14 даув Mgs. 19 ? ? — Abundant 0 3.14 187 (а) + ++4++ | +++-+ 0 0 10.14 55 0 0 ++ Abundant 0 ). 16 209 ++ ++ | ++ ++ 0 0 ).92 C ++ +++ пат 0 0 10.62 94 qup ++ ++ | ++ ++ 0 0 2.0 204 ? +++ FEF 0 0 ).16 220 T 4 ++ Abundant 0 ). 15 ? " ? Abundant | Abundant 21 19 ES ++++ | ++++ 0 0 10.( 209 + +++ I---- D 0 0 .‹ 27 ? ? + 0 0 10. 215(4) +-+ +++ +++ 0 0 2 П ++ +++ +++ | Abundant 0 10.( 216 +++ Tor ++ ++ 0 0 .( 144 у Т ++ 0 0 10. 16 ? ? - + Abundant | Abundant : 103(4) ? ++++ | ++++ 0 0 zo 226(B) F + ++++ 0 0 . 04 188 ++++ | ++++ | ++++ 0 0 > 86 "e pF | -++++ 0 0 11.4 97 Нея an "PEE ЕЕ 0 0 ).: 95 IPES 3papar +++ 0 0 а В ? ? + Abundant 0 10. 7 ? + 4-4. 0 0 Г. 48 ? + +++ | Abundant 0 10.: 11(2) +-+ рег ЗЭЭР 0 0 8. 56 ? + ++ 0 0 9. 5-2 0 ? + Abundant 0 4 194 0 + “ер 0 0 б. 185 + + +++ 0 0 „08 165 ? + pata sae 0 0 10.06 25 ? +++ p 0 0 .94 44 0 0 йн Abundant 0 .46 1 0 + TET 0 0 .66 218 2 +++ лт 0 0 3 HI ar +++ ама ЈЕ 0 0 6.! 215(2) ЗЕ +++ estate 0 0 6.1 78 0 ? A 0 0 10.6 198(1) T +++ +++ 0 0 8.7 15(1) 0 0 -- 0 0 LA Control 0 0 0 Abundant | Abundant ——— Control 0 0 0 Abundant | Abundant —— Control 0 0 0 Abundant | Abundant — mgs. of dextrose ог 100 to 120 parts of sugar consumed for each part of nitrogen fixed. This would indicate that the organic [Vor. 15 148 ANNALS OF THE MISSOURI BOTANICAL GARDEN food available, provided the particular culture in question is capable of utilizing it, is possibly of more significance in determin- ing the quantity of nitrogen fixed than is the culture. There was, however, a rather marked variation in the rate at which various cultures consumed the dextrose. Another very evident fact is that the quantity of visible growth cannot be taken as necessarily indicating the relative fixation of nitrogen. Cultures Nos. 219, 55, 220, 27, 144, B, 44, and 178 all showed a relatively small visible growth, yet they were among the most active nitrogen-fixing strains. Cultures Nos. 15 and 16 were evidently unable to utilize dextrose very readily. It is possible, though, that they may have fixed as much nitrogen per unit of dextrose consumed as any other culture. From the cultures tested in the experiment just described nine, Nos. 7, C, 165, II, 188, 194, 187, 178, 218, and in addition R, were chosen for a comparative study of their ability to utilize the various acids studied in previous experiments. These particular cultures were selected both because of their active nitrogen-fixing ability and because they were secured from such widely varying conditions. Furthermore, they exhibited rather marked variations in cultural characteristics, indicating that they represented different strains or possibly species. In this experi- ment 100 сс. of the medium were placed in 750-сс. or 1000-се. flasks, thus giving excellent aeration. The rate of visible growth is indicated in table ххтх, while the quantities of acid consumed and nitrogen fixed are recorded in table xxx. The data presented in tables xxrx and xxx tend to confirm the previous results secured from cultures No. 62 and 94 in that rather marked variations are exhibited in the ability of various cultures to grow in the presence of calcium salts of different organic acids as the only organic matter present. All ten of the cultures readily assimilated dextrose, though culture No. II probably with somewhat more difficulty than the others. All grew in the presence of the calcium salts of acetic, propionic, normal valeric, normal butyric and normal caproic acids, the latter apparently being more easily metabolized than TABLE ХХЇХ RELATIVE ABILITY OF TEN DIFFERENT CULTURES OF AZOTOBACTER ТО GROW WHERE THE CALCIUM SALT OF 1928] DIFFERENT FATTY ACIDS WAS THE ONLY SOURCE OF ORGANIC FOOD GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 8 |++++ +++ | +++++ cDo e Saa PHHH | ЕРЕ dH 44444 $ [EHEHE IE | а | | ++ В |++++ +++++ | +++++ E. 485 о 32 |++++ ‚++ +++ БЕЛ Банан ка | АЕ +++ | ttt See | +++++ +++++ | 222 | АЕ ТЕ до |+++++ +++ ++++ 45 (27 ++ 888 осо-444 | оооо+ |с-с-с-с-с. ortig | ome ++ iE $ ЕЕЕ 8 | 14 +++ $ | 3.8.8 | ++1 TT |еосо~ |е-е-е-о-е- | +++ | ~ +++ 5| 38 5 » ЕЕЕ Бет ри НЕ de 4| ESS еее +++ | ++ | T+ +++ PI^" |++++ ++ + 5 = 855 с-0-0-0-0- | Oe c.c dE | сс-с-с-с- | оо +++ | occ ++ 2 БЕ + ++ | +++ ЕЕз Ж % Е + ++ LE = ++ + | 88 | ЕН | +++ | +++ | +++ +++ E UL + ++ +++ 55 | +++ +| ++ $8 көкет енін ЭЭН 13011 44417 47 +++ + +E +++ Р о је " о % 3 N — TABLE ХХХ (continued) [Vor. 15 150 ANNALS OF THE MISSOURI BOTANICAL GARDEN 7 ұза | tttet ан ү++++ | ++++ В а НЕЕ b € B BB ово за гени 2 Typ HEHHE] +++ | P4444 | 13444 А +++ | +++++ | ++++ | 1++++ 2 СРЕ сосос | оооро | ососос | ООСООО | оооое e e ЕС + +++ | 4| elit ааа | АРА атана +++ lii 28 ++++ | +++ |+++++| + +444 5 (38 TT HES | ссесе | + + ын ESE +++ Г-1-1-1-1Ё | с-с-с-с-с- | с-с-с-с-с- 4d 5 |—5 ТЕЕ +++++ + m са 3 » РЕ + +++ ++ а | 5 ++ +++ | + БЕ ++ | +++++| 4 +1+++ 5 | ESE |е +++ | +++ +++ |+ 8 | 54% ++ | +++++ | t+ ннен 5 | 7 tF +| +++ мал +++ 5 „© BER | өвсөн eedi|eeees |+++++|+++++ 2 74.9 + + ВЕЕ ооо | +++ «нэн Л Л +++ | +++++ == + + Zo + = a 8: eut * ++ + EE әсе | +++ | +++ | +++++ | 44447 +++ + gg + +1 .+:+ ++ 88 | 44-17 |-44--1 | +++ | | +++ 5% ЕН | ++ H ++ is 22295 |22255 | SESTS |12055 2332 H g i Б E (d) Observations made after 23 days incubation at 30? C (e) Observations made after 25 days incubation at 30? C. C C. C. ade after 13 days incubation at 30* * (a) Observations made after 5 days incubation at 30° b) Observations made after 8 days incubation at 30° (c) Observations m ( 1928] 151 GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 818 91 62€ Yo— 19 398 OL 802 168 626 811 818 с 568 2 гад! 816 L¥Z 681 986 986 281 818 LYZ 19— LL OLE 62 682 179 989 РӨТ 818 y 126 се #8 118 2%6 989 129 089 881 829 ST 291 L¥ 911 975 66 192 785 866 П 818 6 62€ Ұс да! PLE 22. 296 779 1434 991 818 82 096 EFI 921 све СУТ 895 299 968 812 818 486 9T 78 198 тет 916 ole 268 о 818 ST 296 91 991 PLE 101 008 979 788 L 818 ! 866 ££— 281 8168 ША 945 819 288 Ч peurnsuoo prov 'S2]A Tr ) pee 4 T qe pp - T qo Ж-Е 821 прерије 0 "чер 4 т ЧЭРЭР + Tek Ж qo 181 PEET ) TPT + 4 т РЕЧЕ T ЗЭР ET ЗЭЭЭР 761 TELT ) е збе а У и ж ere ee che - ++ МОРЕ ЕР" TEF 88T Ter ) + 0 4 - 4 + a zd = II qp e ) Tope 4 T qq T МР 98,288 ая тя 991 debet 0 аки “эр фе н + пре G ^. эри жишш май дин ни зи а 816 T4 0 TET 4 4 тр 4 ее ET +++ о TT 0 ЗООГ v é и им и + ++ те | TFT 2 «ait ( лерд + d алы жа. ін 2 + db Ше ЭРЭЭН Ч (uonsqnout вХєр Gg) цумола o[qIST A тов 108 тов OLI9[UA |DIO€ ошојва тов тов тов оволјхо(т Заек У сени Du fees ший: алаад ва м ? proe "ON -081 [BULLION -L -очой [1810 № -ов| puy | 771016944 | опеоу это HHLOVH -OLOZV чо БЯЯЛАЛЛО гхячяяаята NAL АЯ АЯХІЯ NS ODOSLLIN (NV ватоу ALLVA 6АаОГЧУА AO NOLLVZI'ILLO 'HLAOSS XXX WIgVL (Vou. 15 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN 152 егі СФ 1 na £6 28 1071 EL eL Алотво 194 рэхц пой -O1j вази 73`9 ажи! 88 с 9 96 v 00 9 8976 892 "uid iod pexg пой -ощта Зүү ест 96 LIT 091 202. 191 012. 968 э8влэлу 76 — 96 кар” LL 305 16 181 022 875 821 РОТ -= 201 - £6 FSI 78? 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[шло -081 puy | 2014014 | әцәоу этап (рэпициоэ) ХХХ ЧЛЧУЈ 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 153 any of the others, with valeric ranking next. It is quite evident, then, that increasing the size of the molecule does not decrease the availability. Not one of the cultures seemed capable of growing when the calcium salt of iso-caproic acid was the sole organic compound present. Growth with the calcium salts of iso-butyric or with the two hydrated valeric acids (the only organic materials present) was usually either very slow or absent. Culture No. 188 seemed to utilize a wider variety of acids with more ease than any other, its growth being recorded as + after three weeks with all the different acids except iso-caproic. As previously pointed out, the quantity of visible growth is not necessarily associated with the quantity of nitrogen fixed or the quantity of organic material consumed. This is again evident if the visible growth as recorded in table ххтх is compared with the quantity of acid utilized and nitrogen fixed as recorded in table xxx. The quantitative data presented in table xxx show very definitely that all ten of the strains of Azotobacter used in this experiment can utilize, as a source of organic food for nitrogen- fixing purposes, all the acids tested with the exception of tri- hydrated valeric and iso-caproic. Cultures C and 218 apparently utilized the trihydrated valeric acid. The iso compounds, though, cannot be assimilated as readily as the straight-chain molecules. Since this experiment could not be repeated no effort will be made to analyze critically the results secured with the individual cultures. It is quite evident that some of the cultures can assimilate certain acids very much more readily than can other cultures. Furthermore, some were capable of fixing very much more nitrogen per unit of acid with a given acid than were others. The milligrams of acid consumed per milligram nitrogen fixed, the nitrogen fixed per gram acid, and the quantity of nitrogen fixed per calory of contained energy for the various acids were averaged and recorded at the end of table xxx. In a general way this summary agrees with the results secured with Azotobacter vinelandii. The quantity of nitrogen fixed per gram of acid metabolized increases as the molecular weight increases. In fact the increase in quantity of nitrogen fixed is more rapid [Vor. 15 154 ANNALS OF THE MISSOURI BOTANICAL GARDEN than the increase in heat of combustion, indicating a more efficient use of the energy contained in the larger molecules. 'The conclusion seems justified, then, that the calcium salts of acetic, propionic, normal butyric, normal valeric, and normal caproic acids can be very readily assimilated by numerous strains of Azotobacter, while the ability to assimilate iso-caproic acid is rather limited among these organisms. The other acids tested, namely, iso-butyrie and iso-valeric acids, while capable of being assimilated by most of the cultures tested, are certainly not as readily available as are acetic, propionic, normal valeric, and normal caproic. As a further check on the relative availability of the various acids the protocol arranged in table хххг was carried out experi- mentally. Тһе acids used in this experiment were all from new lots and, because of limited quantities available, only approxi- mately 0.5 per cent concentrations were used. Even with 0.5 per cent, large quantities of heptylic and caprylic acids remained undissolved in the culture medium. Erlenmeyer flasks of 300 сс. capacity containing 50 сс. of the medium and inoculated in duplicate served as cultures. In addition to the fatty acids a number of polyhydric alcohols of varying molecular weights and some of identical molecular weights but varying configuration were used in order to see if similar variability in assimilability, as observed for acids, existed for alcohols. Culture No. 94 again demonstrated its superiority over either No. 178 or No. 218 to utilize a variety of fatty acids, and also to metabolize more readily the alcohols. Both the latter cultures again failed to utilize any of the iso compounds readily and some not at all. These data, secured from entirely different batches of acids, tend to substantiate the observations noted from experi- ments recorded earlier in this paper. One new acid, di-methyl- ethyl-acetic, was added but apparently was not even assimilated by culture No. 94. The same variability as regards the utilization of different acids by different strains of Azotobacter is also evident when the alcohols are considered. Of the eight polyhydric alcohols tested, only sorbitol and mannitol were readily metabolized by 1928] 155 GAINEY— SOURCES OF ENERGY FOR AZOTOBACTER 0 0 0 0 0 0 0 81419 го 0 1023100) 0 0 0 0 0 0 0 0 + 0 0 0 1023002) +++ ++ | о +++ ++ +++ +++ 10414106 Чет 4 0 0 0 0 0 ОРАН ЕР ТРЕВИ АН + 103600] 0 0 0 0 ) 0 0 0 ЈН У | НЕ ++ +++ 10343 Ary 0 0 0 0 ) 0 0 0 + 0 0 0 Тодора] ++ + ++ | 0 0 |++++++| + ро |++++++++++++ гоугаавуү 0 0 0 0 0 0 0 0 + 0 0 0 [oyruopy +44 + | өе 0 0| ео Of јетре Ре LT (922413) 0 0 0 0 0 0 0 0 вл 0 0 0 [094[2 ouq} 0 0 0 0 0 0 0 0 0 0 0 0 prov оп9оз-14ч9-Афешча Eua [44 | 9 ннн и ннан наннан Боз oiadoq ао P T T T Џ Џ Т Uu S fa te be = 489-08 phat ЈАРЕ кен жек е FE HH HER prov 2002450 [euo TEE 4 0 4 0 0 0: РРА ++ 1 + 0 plow опә[вА-ов] ча + ++ [++++++ + | + +++|+++ ртов опәүзА решаок к П + 1-1 T га è Па 07 Ж НЕЕ 1 2 +2. ЫРЫ 108 ален, ELM TER У ЕАУ ЕАУ А ++ Li 4 сав en Е T modem pio 01010021 +++ ++ | ++ | + |+++++++++| ++ ++ +++ prov опеоу 0 0 0 0 : 0 0 0 0 0 0 0 plow oruwo sXep | вАвр | ввр | вавр | s&ep | sXep | звр | ввр | s&ep | вдвр | вХєр | вдвр 812 ок emn) SLT ‘ON omn; $6 ок әлің втоноопу аму ватоу OINVO3O влотчуд AZITILA OL 816 ANY '821 96 "SON SHUNLINO UALOVAOLOZV AO ALITIAV AHL AO 616351, WAILVIIIVAO IXXX WIHV.L Мог, 15 156 ANNALS OF THE MISSOURI BOTANICAL GARDEN all three cultures, the former more rapidly than the latter. Dulcitol, an isomer of sorbitol and mannitol, apparently cannot be utilized by any of the three cultures. Inositol, a six carbon hexahydrie ring alcohol, was readily utilized by culture No. 94, with more difficulty by culture No. 218, and not at all by culture No. 178. The same was true of glycerol. The four carbon alcohol of this series, erythritol, was utilized only by culture No. 94. Тһе two and five carbon numbers, ethylene glycol and adonitol, were apparently not available to any of the three cultures. This fact is rather interesting since Mockeridge obtained the maximum fixation in her experiments where ethylene glycol was the sole source of energy. Apparently the configura- tion of the molecule plays an important role in determining the ability of a given strain of Azotobacter to utilize a compound. DISCUSSION Attention has already been called to the salient facts brought out in the various experiments in the summaries accompanying the individual tables. In the limited discussion to follow an effort will be made merely to correlate these various points with the special object of trying to see if satisfactory answers to the original questions propounded in the introduction can be made. In the first place, it is frankly admitted that certain procedures followed in some of the earlier experiments did not prove as satisfactory as had been hoped. This was true of the aeration in that the quantitative acid and nitrogen data did not, for some unknown reason, check as well as did later experiments. Because of these irregularities as much significance is not attached to the data secured in connection with those experiments as to those in which culture No. 94 was employed. Secondly, such variation in the ability of different cultures to utilize different organic food substances was not anticipated or the experiments would have been confined entirely to identified cultures, thereby making it possible to compare results here reported with results obtained by other investigators. This desirability was realized in time to make use of known cultures in the major portion of these experiments, and data obtained with known organisms are regarded as much more significant. 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 157 In the third place the inadequacy of the data, in many or possibly all instances, from a quantitative point of view, is realized. It is believed, however, that in this respect they are equal or superior to any thus far reported. Where quantitative determinations, such as were employed in these experiments, are necessary the accumulation of mass data is a slow process. The limited data available are presented not as definite proof but rather as indicating certain tendencies, and it is hoped that others may see fit to conduct experiments along similar lines, thus bringing about the accumulation of sufficient data to justify definite conclusions. It is desired to call attention again to, and to emphasize, the danger of applying the findings from a study of one strain of Azotobacter to other strains. If there is any one thing indicated by the experiments reported in this paper it is the marked variability in the metabolism of organic compounds of different strains or species in this group of organisms. ‘This fact makes it rather difficult to compare results that have been reported from one laboratory with those from another, because frequently no indication whatsoever has been given as to the origin or identification of the culture studied. In the future much greater emphasis should be placed upon the identity of the culture being studied. For the reason just set forth it is believed that very little would be gained by a comprehensive review of investigations dealing with the utilization of various organic substances by this group of organisms. A cursory perusal of the literature dealing with Azotobacter will impress one with the great variety of organic compounds that may be assimilated by these organisms. Also it brings out the wide variation in efficiency with which such compounds can be used when the quantity of nitrogen fixed in their presence is the criterion by which such efficiency is judged. To illustrate the points suggested in the preceding paragraph the reader is referred to table хххи. This table is an adaptation of one recently used by Bonazzi (26) enlarged to include the work of Mockeridge and a few examples from the data previously presented in this paper. Over fifty separate and distinct organic compounds are here [Vor. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN 695 14309-08] 20'6 144044 20% 0071 14435 01:2 0070 Ime tL OI 102418 ƏUA ÁH 00'9 ОР с 9079 8971 SLT TOo199A[D) 88-Р 0070 109443 АТ 69 11 0/79 OF FI 86 Р 96 6 OF 6 To1ruu'e pq :5тоцооту 826 09'0 00'01 FEL 09 1 әвотпавгу 0677 овопивуј 0076 об р OF 6 ус 6 овојАХ 98'8 469 SS'I 46 8 29 1 95 р 09 Р 9503 хэ 05`9 19 0 86 1 911 12272 98 1 96030в[80) 28 OI 24 8 08:01 89 1 088 се 8 эво ләт 6676 080 02 1 79°? 88 8 СТ 6 950398] 992 08 с 740 98 1 ҰР 2 ЭВОЗЕ Р ве 4 00'0 OL ITI 860 06 € 09'8 9501845585 09 1 се 6 26 6 E :влвапа ere 4398288813 um) t1 9 214919 umi) :=шпо 92 6 08'9 cs OI сет 848 1 ЧАЙ: опо] 2979 0071 OF ЕТ out 84 1 SIL sult} хэ 86 € OF 0 0676 ЈЕ 6 191835 :воршвцоосввАјод «9104 әлү einq 9204 einduly :0080 – #0080 + Keurer) |одршолоор | Aysurery ~ nte M gm шттреш ut punoduioo 2108310) [91193€$ur orueZ1o ure1d 194 рэхц подолуш ‘58ү 158 SHOLVOLLSHANI гхянялата AH аялчнявяо аООЯ OINVOHO иучо H3d N3DONJLIN ЯО NOILVXIA ЯАТІУТЯН IIXXX WIHV.L SOURCES OF ENERGY FOR AZOTOBACTER 159 GAINEY 1928] "(punur 42)2000)02 V) $6 ‘ON 9апупо тор IIB вувр 19430 "tpupjeuia 1грофоюгу PUB 1220200442 42912040102 Surpn[out воли по Jo 1equinu э818] шолу дувипорво 10} BIB] ‘(99220200442 1дтфоюгу) 09 ок 91nj[nO чим PINs esorjxop 10) 83801 , er'G 93881-80) 811 9181|034(8-82) 88 I ә}вәүвш-вгу 00 6 эувг80101-82) 6, 9 93391-80) 264 зувпорвш-во) МГ“ 93801998 1-80) ҮС Р 9]91)19]-9/) 28 6 90 6 9]8118]-EN, 96 0 2170 9)8|8Х0-3 10721 се 6 бӯ С авроврво ҰР 9 09 1 00 I ест 918:112-8Х| 09 8 9увп100п8-в() 2872 962 әұвшәәпв-е № :вртов 2тив210 19410 JO 81188 18 1 (ost) аувол9во-в) 1821 978010 80-89 69 A (раувар чи) әўвтләүвА-вгу 669 (роуварХцопош) әўвтлә[вл-®гу 14 ОТ (1вшлоп) әўвтлә[вл-®С) 8976 (ost) 9)8:41114-82) 2479 80 9 9170 2070 э)814)14-82) 99 3 919 00'0 ejeuordo1d-e 96 0 0171 оувпотдола-вМ 09 2 118 02 6 оувудов-во) 00 0 Т 9]'€ULIOJ-9/) :вртов 4339] JO 81188 „ә әл әл эта олпа шј #0080— | "0989+ ләшш® ndur—re ; Кдшво јодрмолоор | Амвителм pus NE pem umrpeur ur punoduroo 2108810) твидувш orueZ1o ure12 194 poxy подолу ‘SIW (рәпчциоо) [IX XX. ЯТЧУ4, [Vor. 15 160 ANNALS OF THE MISSOURI BOTANICAL GARDEN recorded as being assimilable by Azotobacter. Among these are representatives of classes of organic compounds possessing, in many instances, very few characteristics in common. Further- more, the compounds here listed are only those reported by five of the many investigators in this field and are by no means intended to represent all compounds that have been tested and found capable of supplying the organic needs of members of the Azotobacter group of organisms. Among the carbohydrates are examples of polysaccharides, gums, and sugars. Among the latter are tri-, di-, and mono-saccharoses as well as hexose and pentose sugars. Alcohols are represented by mono-, di-, tri-, tetra-, and hexa-hydrox compounds, also by straight-chain and iso arrangements of the carbons, with the additional variations in polarity of inactive, dextro-, and levo-rotary molecules. There are also twenty-five salts of organic acids, including representa- tives of a number of dissimilar groups. There would seem to be no question, then, but that among the species of Azotobacter there are members capable of utilizing a very wide variety of organic compounds as the only organic requirement for the fixation of nitrogen. Not only is this true but the same strain of organisms can function as a nitrogen fixer when supplied with a wide variety of compounds. When it comes to the comparative efficiency with which these various compounds can be used, measured by quantitative gains in nitrogen, the data are too inadequate to permit of drawing any very definite conclusions. The work with impure cultures would have to be eliminated from consideration. This leaves only a few compounds that have been tested by two or more investigators. Of these ten may be selected that were studied in common by Hoffmann and Hammer ('10), Krainsky (08), and Mockeridge (15). The quantity of nitrogen fixed per gram of material as reported by these investigators is indicated in fig. 3. The data plotted may not be very accurate, since there is no indication of quantitative determinations of the residual organic material having been made, except that Mockeridge states that qualitative tests showed complete absence of the organic com- pound. Furthermore, one is forced to assume that the original 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 161 1.0 per cent of material added remained quantitatively unaltered during the process of sterilization, a condition that certainly might not obtain in all cases. In this connection it is believed that the policy followed in experiments herein reported, of making quantitative determinations on controls submitted to the same treatment as the cultures, is а much safer procedure for ascertaining the quantity of the organie compound available to the organisms. From Hoffmenn+Hammer с т 0 mn From Mockeridge /2 pu —-— from Krainsky mom. nitrogen fixed pr qm material зп чш А о мч d ~ ~ / d 1 / P № шал Р Ж а” \ е-е m NT X 2 — ~ ^ 9 5 ~ ~ я 4 v е o 2 2 35 5 > = з 2 “See 21: ы ~ о + М 5 ж а с о а sS 7%) + A. I d < Е 5 Fig. 3. Showing fixation of nitrogen per gram of organic material, secured b different investigators. In spite of the numerous possibilities for errors the curves in fig. 3 show a marked qualitative similarity. The absolute quantities of nitrogen fixed per gram of material vary very widely. This might be taken to indicate a variation in the efficiency of the cultures, but, as mentioned in the preceding paragraph, it may merely mean that the organic material was not used up quantitatively in, for example, Krainsky’s experiments. If it is assumed that in all instances approximately the same quantity of organic material was available, and that it was used up quantitatively, then we would be justified in concluding that there are wide variations in the efficiency with which different ГУ ог, 15 162 ANNALS OF THE MISSOURI BOTANICAL GARDEN cultures utilize the same compounds and also a marked contrast in the efficiency with which the same culture used different organic materials in the fixation of nitrogen. There are strong indications, however, that different cultures may utilize many of the same compounds with approximately the same relative efficiency. Attention was called to this in connection with the experiment reported in table ххш, in which out of forty cultures tested the quantity of nitrogen fixed, when dextrose served as the organic food, did not vary very widely in most instances. The work here reported has been confined almost entirely to the salts of fatty acids, principally calcium salts. And attention may be called again to the possible marked effect that the cation may have upon the availability of the fatty acids, as indicated in table rx. It is necessary of course to use the salt of any acid to be tested in order to avoid the inhibiting effect of the hydrogen-ion concentration produced by the free acid. The only previously reported work with which these data can be compared directly is that of Mockeridge, recorded in table хххп. That portion of Mockeridge’s data dealing with fatty acids has been recorded in table ххх parallel with a summary of the data presented in this paper. In this table are recorded in parallel columns the molecular weights, heat of combustion, nitrogen fixed per gram of acid consumed, and nitrogen fixed per calory of heat energy contained in the material consumed. Qualitatively, the only point of difference between the data presented here and those reported by Mockeridge is in the utilization of formie acid. In no instance have either quantita- tive or qualitative evidence of the growth of any culture of Azotobacter in the presence of a salt of formic acid been observed in this work. Quantitatively, the data agree in showing that as the molecular weight or heat of combustion increases, the quantity of nitrogen fixed per unit of acid consumed increases. "This is brought out clearly in fig. 4. There is, however, a difference, possibly significant, in the type of curves plotted from the two sets of data as shown in fig. 4 where the nitrogen fixed is plotted against molecular weight, or heat of combustion, since both increase arithmetically in the compounds as arranged in the figure. ад Фош УЗАК ЕМА МЕ У, з 1* ALS + Dit? г Оо то”. . 1928] 163 GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 'SanpeA uornsnquioo jo звәц |пәләшір pesn эЗршэхэорү( "548 чт рорлооол uorjsnquioo jo вувец 40) ројајпојвооц 11 'Sspunoduioo ewou 10] вв oures oq 03 решпвву || 'suorjsutuil9jop 8 JO одвлолу 1 ‘ѕиоцешоләјәр е JO эЗвлхэлү | `°вцпот})витшлә1]әр с JO әдвләлу „ Тт 19'9 се 6 98 8 9/76 c 219 081 980.1}X9(T 9071 «21 1 91:14 Не 088 911 101d 89-06 2111 +460 Дд 59 9 iio 189 сот эшэ|вА-рэ)варХц-ид, 06`0 56679 8979 1197189 201 още вл-рафвтрАч-опод 0970 «69 € 969 Ну 759 88 о/пд-овт 67 с $28 4I 9r 4 6 '0£8 9IT oro1d'e) 8971 512701 89 9 9 189 201 SUTEA. 20 I 809 наа! 12978 96 9 Р РСС 88 ou&jng РО Т 91 9 YII 1999 467 Ӯ 198 2) отиотдота 60 Т 22 8 740 +09 2 бре ғ 605 09 onoeooy Нотт 4871 0070 0070 те I 2719 ОР OTULIOf эзршэхэор() | (одрмоцоо 01313 194 fl лод P um 227 oa и 22190912) поцвпашоо у ом роу ох ох . 4 uorjsnquioo о |вэ I9[noo[o : ” Кол! Sp uos ont 19 И подолу “Яр | подолу “SBP фази 1 Н mW ЯО АЧОЛУО МЧА аху иучо чяа аяхія кяоочатх ялигулян аху NOILSAAWOO dO LVAH ПІХХХ WIGV.L (ПахутяхтлА ЯЯТОУЯОЛО?У) 76 ОК нмаллпо ха ватоу OINVOHO ‘LHDIGM ЧУПТПЛОЯЛОЙ [Vor. 15 164 ANNALS OF THE MISSOURI BOTANICAL GARDEN This difference is shown more strikingly in fig. 5 in which the nitrogen fixed is plotted against a unit of energy contained in the compound. /8 2 7 № 2108 7 | 7 ~ 1741 — оо № Calories Аеа# рег gram acid — Heat of combustion ~~~ Nitrogen fixed - Gaineysdaty —— Nitro gen fixed-Mockeridge Чи, Тат. nitrogen fixed рег gram acid Acetic Р Ви ty ric Valeric Caproic Fig. 4. Comparison between nitrogen fixation and heat of combustion рег gram acid. If the quantity of nitrogen fixed were proportional to the heat of combustion, then in fig. 4 all three curves should run parallel, while in fig. 5, the two lines should coincide and lie horizontal to the base line. In fig. 4 the nitrogen-fixation curve plotted from Mockeridge’s data tends to diverge from the curve for heat of combustion, and in fig. 5 it approaches the abscissa. Such curves would indicate that as the molecular weight increases the quantity of nitrogen fixed рег unit of energy decreases. Тһе decrease indicated in Mockeridge’s data is very slight and is probably within the limit of error. On the other hand, the nitrogen fixation curve 1928] GAINEY—SOURCES ОЕ ENERGY FOR AZOTOBACTER 165 in fig. 4 based upon new data starts well below the heat of com- bustion curve and actually crosses it with a marked upward tendency rather than a tendency to flatten out, while in fig. 5, starting on the base line, the curve continually, though not uniformly, rises from formic to саргос acid. Such curves indicate that as the molecular weight increases the organism is capable of utilizing the contained energy more efficiently. Atten- 47 7777 From Mockeridge's data —— from Gaineys data Тат. nitrogen fixed per calory Formic Acetic Propionic Butyric Valeric Caproic Fig. 5. Showing fixation of nitrogen per calory heat of combustion. tion has previously been called to the fact that growth has been uniformly more rapid in caproic, and frequently in valeric acid, than in the acids of lower molecular weights. The data serving as a basis for figs. 4 and 5 were secured from normal acids. In no instance has the iso acid appeared to be used as rapidly as the corresponding straight-chain compound. No explanation is offered as to the reason for this unless it is [У отг. 15 166 ANNALS OF THE MISSOURI BOTANICAL GARDEN that the straight-chain compounds are more frequently encount- ered by the organisms in nature and as a result they have become better adapted for metabolizing substances of this type. The data presented in table xxxi indicate the same tendency оп the part of Azotobacler vinelandii to utilize more efficiently, though not to the same degree, the iso acids of a higher molecular weight. When the fatty acids are compared with dextrose and their efficiency measured in terms of nitrogen fixed, two points in connection therewith are worthy of note. If the nitrogen fixed per gram of material be used as a basis for comparison dextrose ranks about on a par with butyric acid, acetic and propionic being much inferior, while valeric and caproic, especially the latter, are superior. On the other hand, if the energy content be used as a basis for comparison, then dextrose ranks only slightly below caproic acid. The data presented in table xxxii are inconclusive in showing & close correlation between the energy content or heat of com- bustion of an organic compound and the efficiency with which Azotobacter can utilize it as the organic food required; never- theless, there is more indication of a correlation when compared upon this basis than simply upon the actual weight of the material. There is an urgent need for much more data on the series of compounds herein reported, as well as other series, secured under carefully controlled quantitative conditions with definitely identified cultures, before any very definite conclusions ean be drawn as to the energy relations concerned in the fixation of nitrogen by Azotobacter. SUMMARY 'The experiments reported in this paper have been carried out with the object of determining the relative ability of different cultures of Azotobacter to utilize qualitatively and quantitatively calcium salts of the different fatty acids up to and including six carbon atoms. Two cultures have been studied intensively, others to a less extent. The quantity of acid and total nitrogen present in the medium before and after varying periods of incuba- tion were determined. The rapidity of growth was recorded, 1928] GAINEY—SOURCES OF ENERGY FOR AZOTOBACTER 167 as was also qualitative tests of changes in hydrogen-ion concen- tration. The following is a summary of the more important tentative conclusions indicated by the limited experimental data secured: (a) The various strains of Azotobacter behave quite differently with respect to their ability to utilize different fatty acids. Some cultures have been tested that seem rather limited in this respect, while others may utilize all of the acids tested. (b) Individual cultures may vary widely not only in their ability qualitatively to utilize different acids but there may also be a marked difference quantitatively in this respect. (c) The iso compounds are not as readily metabolized as are the normal. (d) There is a marked tendency toward the reduction of the hydrogen-ion concentration in a medium in which an acid is utilized by Azotobacter. In some instances; apparently, this phenomenon may be responsible not only for the cessation of growth but actually for the death of the organisms. (e) The quantity of nitrogen fixed is more or less proportional to the quantity of acid utilized. (f) The quantity of nitrogen fixed per unit weight of acid consumed increases as the molecular weight or heat of combustion increases, provided comparisons are limited either to normal or iso compounds. There is some indication that the efficiency with which Azotobacter can utilize various acids, as measured by the quantity of nitrogen fixed, increases as the molecular weight increases, even when the comparison is based upon the energy content of the material utilized. (g) The cation with which the acid is combined apparently plays a very important role in determining the ability of an organism to utilize the acid. (h) The quantity of nitrogen fixed when various acids are utilized is more closely correlated with the energy content than it is with the actual weight of the material consumed. ACKNOWLEDGEMENT I wish to take this means of expressing my appreciation to the officials of The Missouri Botanical Garden and The Kansas [Vor. 15, 1928 168 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Agricultural Experiment Station for the valuable aid rendered in making possible this investigation; and especially to Dr. B. M. Duggar under whose guidance the work was undertaken and whose constructive criticism has proved of inestimable value in its prosecution. LITERATURE CITED Bonazzi, A. (26). The mineralization of atmospheric nitrogen by biological means. Conf. Internat. de Pedologie, Actes 3: 74-115. 1926 Hoffmann, C., and Hammer, В. W. (10). Some factors concerned in the fixation of nitrogen by Azotobacter. Wis. Agr. Exp. Sta. Res. Bul. 12: 152-172. 1910. Hunter, О. W. (23). Stimulating the growth of Azotobacter by aeration. Jour. Agr. Res. 23: 665-677. 1923. Johnson, Н. W., and Lipman, С. В. (22). The effect of reaction on the fixation of nitrogen by Azotobacter. Univ. Cal. Publ. Agr. Ба. 4: 397-405. 1922. Krainsky, A. (08). Azotobacter chroococcum und seine Wirkung im Boden. Centralbl. f. Bakt. II. 20: 725-736. 1908. Latshaw, УУ. 1. (16). Sodium sulfate as a substitute for potassium sulfate in the Gunning modification for determining nitrogen. Jour. Ind. and Eng. Chem. 8: 586. 1916. Lóhnis, F., and Smith, N. В. (16). Life cycles of the bacteria. Jour. Agr. Res. 675-702 Mockeridge, Florence А. (15). Soil organic matter as a culture medium for Azoto- bacter. Biochem. Jour. 9: 272-283. 1915. Shaffer, Р. A., and Hartmann, А. Е. (21). The iodometric determination of copper and its use in sugar analysis. П. Methods for the determination of reducing sugars in blood, urine, milk, and other solutions. Jour. Biol. Chem. 45: 365- 390. 1920-1921. THE EFFECT OF ULTRA-VIOLET RADIATION UPON HIGHER PLANTS! ETHEL TABER ELTINGE Jessie R. Barr Research Fellow in the Henry Shaw School of Botany of Washington INTRODUCTION The sterilizing action of ultra-violet radiation has been known for over fifty years. Downs and Blunt in 1877, working with putrefying material, were the first to discover it. Since then there have been many workers in this field, and to-day ultra- violet sterilization is a more or less common practice. At present, work with ultra-violet rays is carried on along two different lines. One line deals with the effects produced in higher animals and man with particular reference to the depth of penetration of the rays and to the changes produced within individual cells. The other line deals with the effects produced in plants. Little work was done in the latter subject until 1911 when Kluyver studied the effect on plants of a long-continued raying with an ultra-violet lamp. From then until 1918 the subject received little attention. Since then, however, it has taken on a fresh impetus, and to-day there are many people working in that field. At the present time it is generally known that raying with an unscreened quartz mercury lamp causes injury due to the presence of the short rays. The important line of research now is to determine the effects of the longer ultra-violet rays on the different groups of plants, and this can be done only by the use of specific screens to eliminate certain rays. Under favorable conditions the spectrum of sunlight contains rays as short as 291 џи. Thus if a mercury vapor lamp is screened to absorb all rays shorter than 291 џи, the same type of rays penetrate as are found in sunlight, the only difference being that ! An investigation carried out at the Missouri Botanical Garden in the Graduate Laboratory of the Henry Shaw School of Botany of Washington University and submitted as & thesis in partial fulfilment of the requirements for the degree of doctor of philosophy in the Henry Shaw School of Botany. ANN. Mo. Вот. Саквр., Vor. 15, 1928 (169) [Vor. 15 170 ANNALS OF THE MISSOURI BOTANICAL GARDEN the ultra-violet rays are much more. intense, since the atmo- sphere screens out much of this group of rays originally in sun- light. History THE EFFECT OF ULTRA-VIOLET RADIATION ON LOWER ORGANISMS Many articles have been written on the effect of ultra-violet rays on bacteria. Potthoff (20) found that a suspension of bacteria 314 mm. thick placed 151% ст. from the light gave the following results. LENGTH OF TIME NECESSARY FOR THE DESTRUCTION OF BACTERIA Spores Vegetative cells В. anthracis 5 minutes 15 seconds В. subtilis -9 minutes 2 minutes В. mesentericus 5 minutes 1 minute In pigmented forms a short exposure inhibited the production of pigment, but upon repeated short exposures the pigment again appeared. Mashimo ('19) found that the rays most effective in the de- struction of bacteria were those between 295 and 186yy. He proved this by using in a quartz spectrograph a culture of bac- teria instead of a photographic plate and noticing the region where no growth appeared. Bazzoni (14) found that the destructive power of ultra-violet radiation in relation to bacteria increased rapidly with a decrease in wave length, but that this effect was in some way dependent upon association with longer wave lengths. Wave lengths of from 220 to 225 yu killed the bacteria after several hours, while the same intensity of light containing full radiation destroyed them very rapidly. Burge ('17) has proved that ultra-violet rays kill living cells such as bacteria, not by destroying the intra-cellular enzymes but by coagulating the protoplasm. For his work he used bacteria that liquefy gelatin and found that the organisms killed by ultra-violet when ground with sand produced as much lique- faction as ground living organisms. Green (97) found that the destruction of diastase in a leaf was less than in an extract of malt or saliva and concluded that 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 171 either the chlorophyll or the proteins of the protoplasm must act as a screen absorbing the injurious rays. Tanner and Ryder (’23) have found that yeast cells are almost as susceptible to ultra-violet radiation as bacteria, although pigmented yeasts are more resistant than white ones. Nadson and Philippov (’28) used a Bach model of a quartz mercury vapor lamp giving rays as short as 220 uu. Twenty- four-hour cultures of Saccharomyces and Mucor of different species on nutrient agar were rayed at thirty cm. from the light for ten to twenty minutes. For raying, the cover of the petri dish was removed and replaced by a piece of heavy glass with a circular opening in the center. After several days the region where there was no glass was devoid of growth, but just at the edge of the opening where only the slanting ultra-violet rays and hence the long ones were received, there was a marked increase in growth. Growth under the glass was normal. With yeast, not only increased, but abnormal, budding was noticed in the region of increased growth. With some fungi, asexual reproduction was increased while with others it was the sexual. Larger organisms have been used for determining the effect of ultra-violet radiation on individual cells. Barr and Bovie (’23) used amoebae that had been cleared through starvation. They found that after an exposure of three-fourths of a second the amoebae ceased to move and after an exposure of one minute they were killed. At first the edge of the organism was irregular, but in a few seconds it became smooth by swelling. If irradiated for three to four minutes the animal swelled and clear spaces appeared between masses of protoplasm. Soon, however, crenu- lations were present about the border of the organism, giving the appearance of a loss of solution from the inside. Tshuhotine (’23) thinks the rays first affect the plasma mem- brane, increasing permeability by coagulating the colloids. Then the surrounding medium enters and precipitates the protein colloids in the cytoplasm which surrounds the colloid particles of lecithin. Soon the base, coline, is formed which increases decomposition, giving OH ions which promote imbibitional swelling of the protein colloids of the cytoplasm until the cell is completely decomposed. [Vor. 15 172 ANNALS OF THE MISSOURI BOTANICAL GARDEN Brooks (26) found that the shorter the rays, the greater the amount of 2,- 6,-dibromo phenol indophenol penetrating cells of Valonia. THE EFFECT OF ULTRA-VIOLET RAYS UPON HIGHER PLANTS Bailey (94) was the first to notice the harmful effect of light on plants. Не used an electric arc light and found that if a piece of glass were placed between the light and the plant the injurious effects were modified. He found lettuce and radishes very sensitive to the arc light. A few hours raying caused leaves of Coleus to become shiny and lose their purple color when that color was present only in the upper epidermis. When cross- sections of the leaves were examined, the epidermis was found to be collapsed and opaque, coloring the leaf brown. Professor Rowlee, working with Bailey, concluded that the palisade tissue absorbs a large amount of water from the epidermis, due to greater protoplasmie activity, and the epidermis thus emptied соПарвев. The next person to do any extensive work оп plants was Kluyver (’11), who, using a quartz mercury lamp giving rays of 230 uy and shorter, gave the plants one long exposure. He verified Bailey’s results as to the injury produced in higher plants and its modification by using a screen of thick glass. Only the epi- dermis of leaves was found to be affected, but in roots and stems the injury was deeper. Anthocyanin was again found to be decomposed by the short rays which do not penetrate. The longer rays were found to have no effect on anthocyanin. Ursprung and Blum (17) used a new method for determining injury. After raying plants the desired time the cells were plas- molyzed in sugar solution and then deplasmolyzed if possible in clear water. The less the injury the greater was the per cent deplasmolyzed in water. Epidermis and cuticle were found to exert a little protection. Usually cells containing chlorophyll were more resistant than those lacking it. Diatoms were found very susceptible, due to the large amount of silica in their walls. Stoklasa (11) found that a long exposure to ultra-violet radi- ation injured the epidermal cells but did not harm the chlorophyll in adjacent cells. Etiolated seedlings turned green іп two hours 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 173 upon exposure to rays of from 400 to 300 uu, while it took six hours to produce the same result in sunlight. Schanz (20) found that when the rays below 320 ци were cut off the larger part of the red color disappeared from red-leaved lettuce. In like manner he caused the leaves of copper beech to become green. Sheard and Higgins (27) reported the effect of ultra-violet radiation on germination and growth of seeds. They used an unscreened quartz mercury lamp and screens of ultra glass, vita glass, and ordinary glass. In general they found that wave lengths of 270—320 uu delayed the time and lessened the rate of growth, probably because of changes which carried to their extreme eventuate in the coagulation of the seed albumin. Rays of 320-390 uu were particularly effective in promoting growth. When seedlings of lettuce, radish, and turnip were irradiated one, two, five, and ten minutes, those which normally germinate and grow in darkness showed most rapid germination and best growth when not rayed. Minimum growth was found in seed- lings grown in diffused light. Radiation of these seedlings for two to three minutes by а quartz lamp accelerated the germina- tion and subsequent growth as compared with non-rayed seedlings under similar conditions. Thus they state that raying with the near ultra-violet region aids germination and growth of a cell or normal functioning of an organism which is kept under unphysi- ologic environment. Russell and Russell (27), using а Hewittic mercury vapor lamp, found that when etiolated mustard seedlings were given short daily exposures to ultra-violet rays, dwarfing resulted in direct proportion to the length of exposure. Some chlorophyll appeared in all rayed seedlings. In seedlings grown under normal daylight conditions the dwarfing was not as great. Dane (727) found that soybeans irradiated by ultra-violet rays were dwarfed and the leaf and stem tissue brittle and stiff. Stems of irradiated plants were 11% times as great in diameter as those of control plants. Rayed stems were hollow and showed reduction in medullary rays, Ше meristematic tissue thus remain- ing active for à much longer time than that in control plants. The ordinary parenchymatous cells of the medullary rays had developed into xylem and phloem. [Vor. 15 174 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Beeskow (27) found that a daily irradiation of more than 1% minute caused injury to soybeans, but that irradiation of 1% minute caused no injury and might stimulate growth. When corn plants were rayed they showed an increased calcium and phosphorus content. McCrea (’27) grew Digitalis purpurea to the ten-leaf stage in a greenhouse glassed with vita glass. She found greater growth and darker color than in control plants. The plants were then put outdoors and when cuttings were taken in August and Sep- tember, the rayed plants were found to contain an increased amount of digitalin. Delf and Ritson (Delf, Ritson, and Westbrook, 727) irradiated Pelargonium, Coleus, Fuchsia, Abutilon, Salvia, and Trifolium for various lengths of time and found retarded growth, delayed germination, retarded flower formation, and leaf fall. In addi- tion there was a loss of anthocyanin by Coleus and in many cases a deeper green color produced in Coleus and other plants. Біх- weeks-old seedlings of Trifolium when rayed one-half minute daily showed increased growth. Westbrook (Delf, Ritson, and Westbrook, ’27) used different lengths of days in addition to short exposures to ultra-violet radia- tion. Іп all cases injury was greater the shorter the day. The injury consisted in the development of thinner leaves with more compact mesophyll and smaller and fewer air-spaces, reduction of mechanical tissue, and collapse of the cells of the upper epi- dermis followed by a withdrawal of the chloroplastids from the upper ends of the palisade cells. Tsuji (18) obtained increased growth and a higher percentage of sugar in sugar cane grown in sunlight and rayed daily with a weak ultra-violet lamp. When pineapples were grown in sun- light plus a daily raying of forty minutes, the pineapples were sweeter, juicier, and larger than normal. When banana leaves and stalks were exposed to ultra-violet rays after being cut they kept fresh longer than similar leaves and stalks not rayed. Clement (26) has found that apples rayed for three hours showed a slight yellowing of the green side, and that when these apples were stored the rayed sides did not regain their green color but remained turgid longer than those not rayed. 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 175 Nadson and Rochline-Gleichgerwicht (28), using а Bach model of an ultra-violet lamp emitting rays down to 220 uy, have found that ultra-violet rays cause crystals of calcium oxalate to form in the cells of Elodea densa, Elodea canadensis, and Pterygophyllum hepaticaefolium. These plants were barely covered with water and placed thirty cm. away from the lamp for ten to thirty minutes. The crystals began as small granules, with a chloroplast often as the center, and increased to good size. After two to four days they dissolved simultaneously with the death of the cell. If the cells were treated with a narcotic before raying no crystals were formed. THE EFFECT OF ULTRA-VIOLET RADIATION UPON ORGANIC MATERIALS Calabek (27) determined the effect of ultra-violet rays upon the swelling of biocolloids such as agar. When agar discs were rayed a marked decrease in swelling resulted. It was found that the effect of raying could be preserved in dry agar for several months even if the agar were redissolved. As a result the hypothesis was advanced that the effect of ultra-violet rays upon plants is due to a lowering of the swelling capacity of protoplasm and cell wall in the upper cellular layers of the plant. Hess (26) and others have found that when foods are rayed they are rendered rickets-protective. In vegetable foods phytos- terol is activated while in animal foods it is the colesterol that is acted upon. THE PENETRATION OF ULTRA-VIOLET RAYS Several authors including Henri (12) have found that the depth of penetration of the shorter ultra-violet rays through the skin is not more than .1 mm. However, for this work dead skin was used. Macht, Anderson, and Bell ('28), using living anesthetized animals, have found that with an exposure of one minute ultra- violet rays as short ав 302 uy. penetrate through living skin that is more than .1 mm. in thickness. When an exposure of two minutes was given rays as short as 280 pu passed through. They then tested the penetration of rays into the peritoneal cavity of Мог. 15 176 ANNALS OF THE MISSOURI BOTANICAL GARDEN rabbits and found that with an exposure of two minutes, rays as short as 313 џи penetrated into the cavity. Next they com- pared the penetration of living skin with that of dead skin and found the former much more penetrable. When the dead skin was treated with a lipoid solvent it became as transparent to ultra-violet rays as living skin. The pigment in the skin of a negro was found to absorb almost all the short rays. The same result was obtained by injecting a rabbit intravenously with 1 per cent eosin solution. STATEMENT OF THE PROBLEM The problem in this paper is to determine the effect of ultra- violet light as a whole on higher plants, and whether the longer ultra-violet rays stimulate growth in higher plants. MATERIALS AND METHODS EXPERIMENTAL The lamp used for this work was an air-cooled Uviare quartz lamp from the Burdick Cabinet Co. In the experiments desig- nated as Series I this lamp was used without a screen of any kind. When used in this way the rays given off range from 578 to 200 uy. (5780 А. 0.–2000 A. U.) (fig. 1). When a screen of vita glass from the Hires Turner Glass Co. was interposed between the light and the plants the rays had a range of 578-289 ци (5780 A.U.—2894 A.U.). The experiments using this screen constituted Series 1. A screen of quartz-lite glass from the American Window Glass Co. interposed between the light and the plant permits the passage of rays ranging from 578 to 313 uy (5780 A.U.-3136 A.U.). Exper- iments using this glass are described in Series ПТ. The ultra-violet rays produced by a quartz mercury lamp may be divided into two groups, first, the abiotic rays (short rays), with wave lengths ranging from 185 to 290 uy, which are reducing rays and hence killing rays, second, the biological rays (long rays) which range from 290 to 400 д. These are oxidizing rays and hence stimulating. The abiotic rays being very readily absorbed by the atmosphere are never present in sunlight when it reaches the earth, and were essentially eliminated where either of the 1928] ELTINGE— EFFECT OF ULTRA-VIOLET RADIATION 177 glass screens was used. The lamp was used at 50 and at 100 inches from the plant both with and without screens. Although the atmosphere between the lamp and the plant absorbs some of the short rays, the distance of 100 inches is not sufficient to absorb all the short rays, and 50 inches without а screen allows a large percentage of the short rays to reach the plants. When a lamp screened by vita glass is used at a distance of 50 inches from the plants most of the short rays are absorbed, but none of the long ones. The same screen used at 100 inches ШЕП | Quartz Ground glass Ordinary glass Glass of photographic plate Vita glass Quartz-lite glass Fig. l. Showing the spectrum of the different glasses. from the plants allows only a large percentage of the long rays to reach the plants. When a screen of quartz-lite glass is used instead of vita glass, plants at a distance of 50 inches receive all the long rays and no short ones. The same screen at 100 inches allows only a part of the long rays to reach the plants. Except for one group of experiments mentioned later the ex- posure began with 30 seconds the first day and each day was increased that amount. АП plants under experiment were moved about each week in the greenhouse to eliminate all differences in environmental conditions. (Vou. 15 178 ANNALS OF THE MISSOURI BOTANICAL GARDEN The plants used were as follows: Lactuca sativa L. var. “ Black- seeded Forcing” ; Raphanus sativus L. var. “Early Scarlet Turnip White Тір”; Cucumis sativus L. var. “Improved Green Hybrid”; Ipomoea Batatas Poir.; Phaseolus vulgaris L. var. ''Stringless Green Pod"; Nicotiana Tabacum L.; Coleus Blumei Benth. var. Verschaffelttt Lem. and vars. “Spotted Gem," “Defiance” and “Trailing Queen"; Bryophyilum pinnatum Kurz.; Zea Mays L. var. ‘“‘Stowell’s Evergreen." Cuttings of Гротоеа, Coleus, and Bryophyllum were made and rooted in sand. They were then planted in rich potting soil in 3- and 4-inch pots. Аз soon as the plants had recovered from transplanting they were put under the conditions of the experi- ments. Seeds of Raphanus, Lactuca, Cucumis, and Nicotiana were sown in flats. As soon as the plants appeared above ground they were transplanted to 3-inch pots and put under experiment. Seeds of Zea Mays and Phaseolus were germinated between moist filter-paper and planted in 3-inch pots. All plants when not being rayed were kept under usual greenhouse conditions. A record was kept of the humidity and temperature. ANATOMICAL METHODS At the end of four weeks samples of the leaves of the plants rayed without a screen were taken for anatomical study. At the end of eight weeks samples of leaves and stems of all plants were taken for the same purpose. Care was taken in the sam- pling to take portions which were in corresponding positions on the plants and otherwise as nearly equivalent as possible. The leaves and smaller stems were killed in medium chromo- acetic killing fluid and imbedded in paraffin. The larger stems were cut free-hand while fresh. Two stains were used: Haiden- hain’s Iron Haematoxylin and Safranin-Delafield’s Haematox- ylin. PHYSIOLOGICAL METHODS The rate of chlorophyll decomposition under the different screens, in sunlight, and in diffused light was tested, using an 80 per cent alcoholic solution of chlorophyll in vitreosil test-tubes. The Рн of rayed and control plants in Series I was determined by the colorimetric method. The plant material was pressed in 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 179 a mortar and filtered through cotton. Then after diluting 1 to 10 with distilled water the chlorophyll was removed by filtration through an atmometer cup. The indicator was then added to the diluted juice freed from chlorophyll and the result compared with standard tubes. Starch Storage.—Stem sections were made from fresh material of Coleus and Phaseolus in the different groups. These were stained with a standard iodine solution to show the distribution of starch. Dry Weights.—For determination of dry weights plants were dried to constant weight in an oven run at 1109 C. Ash Determination.—Three grams of dry leaf material of the different plants were put into weighed crucibles and burned over а bunsen burner until a large part of the carbon had disappeared. То finish the burning, the crucibles were put into an electric oven at 600? C. EXPERIMENTAL OBSERVATIONS SERIES I For all varieties of plants this series is divided into three parts, group H, which includes the plants rayed at 50 inches from the light; group F, those rayed at 100 inches; and as controls, group G, the same number of plants not rayed. Series I Н (rayed at 50 inches from the light without а screen).— Six young seedlings of Cucumis were rayed. The first evidence of the effect of ultra-violet rays appeared on the eighth day, when a slightly shiny appearance of the upper epidermis was noted. By the twelfth day there was evident curling of the edges of the leaf. At the end of three weeks the rolling of the leaves was very evident and the lower ones were turning brown and dying. ‘The young leaves never attained as large a size as those on the control plants. By the end of the twenty-ninth day, when the plants received an exposure of fourteen minutes, one to three flowers were present, but the younger leaves were so rolled that the upper surface was hardly visible. When samples of the leaves were taken at the end of four weeks for anatomical study, they were found to be very brittle. The plant as а whole was very stiff and erect. (Мог. 15 180 ANNALS OF THE MISSOURI BOTANICAL GARDEN The control plants (G) had slightly more leaves and were a little taller. They also had several more flowers. Not only were the leaves greater in number but greater in size, being twice that of the rayed leaves. No rolling of the leaves was noted in the control plants. The color of the leaves was the same in both rayed and control plants except in the older rayed leaves which were brownish. Six Гротоеа plants were used. By the sixth day the younger leaves showed a slightly blistered appearance which increased as time went on. By the eleventh day the veins were brown and the larger leaves showed several brown spots which seemed to be more or less superficial. By the end of three weeks the edges of the leaves had turned down. Very few leaves were shed. At the end of four weeks these leaves were also found to be brittle. The control plants again showed larger and more numerous leaves with no browning. In the six Nicotiana plants, the first effect of raying was noticed on the eleventh day, when the margin of the leaves appeared wavy. Ву the thirteenth day the edges were definitely rolling upward. About the same time the upper surface became very shiny, and the leaves were so brittle it was almost impossible not to crack them. At the end of three weeks the older leaves were turning yellow and the younger leaves were so rolled that the upper surface was hardly visible, though no leaves were shed. The control plants showed none of these characteristics, the upper surface of the leaves being very hairy and the leaves larger. The four varieties of Coleus Blumei were put into two groups according to their resistance to ultra-violet radiation. "The group most sensitive to ultra-violet contained vars. Verschaffeltii and "Spotted Gem." At the end of five days a fading of the red color was noticed, and at the end of ten days practically all the red color had disappeared. The glossy upper surface was broken only by the bases of the hairs appearing as dots. The two halves of the leaves were rolled upward toward the midrib and the tips downward, so that the leaves appeared to be only half their normal size and were very brittle (pl. 22, fig. 4). By the end of four weeks all the older leaves had fallen and only a few of the 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 181 younger remained and those were very small and abnormal in shape. The other group containing vars. “Trailing Queen” and “Defiance” seemed to be a little more resistant to ultra-violet rays. Here the first indication of a loss of red color appeared the eighth day. About the same time the shiny dotted appearance of the upper surface of the leaves was observed. The same roll- ing of the leaves was noted as in the other group. ‘Trailing Queen" lost very few leaves even at the end of four weeks but var. ‘‘ Defiance" began to shed its leaves at the end of three weeks. In the varieties of Coleus where any red color was present in the stem a loss of it began to be noted in the tip of the stem at the end of five days and a complete loss at the end of ten days. If after a raying of four weeks these plants were put back under normal greenhouse conditions the red color appeared again to a certain extent in the decolorized tip of the stem and the new growth of stem and leaves was normal. Ten very young seedlings of Raphanus were rayed. At the end of eight days the typical curling upward and shiny appearance of the leaves was noted. Here the rayed leaves seemed to be a little deeper green than the control leaves. At the end of four weeks the leaves and petioles were almost as brittle as the rayed Nicotiana leaves. At the end of eight weeks the plants were so curled they appeared almost dead. The roots of the control plants, as well as the leaves, were much larger, as will be seen in pl. 21, fig. 7 Two sets of Lactuca, containing ten plants each, were used, one set having two leaves and the other nine to ten. The object was to see if the older plants would be more resistant to ultra- violet radiation. The set of plants having two leaves never seemed to get much larger, as will be seen in pl. 22, figs. 2 and 3. At the end of two weeks the leaves were noticeably smaller and fewer than on control plants. All the new ones formed were abnormal and the older ones soon dried up and dropped ой. As time went on the difference between the rayed and control plants became the most striking of any of the plants tried. At the end of eight weeks the rayed plants had an average of 4.25 leaves per plant while their controls had 13. Мог. 15 182 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN This and one set of Raphanus plants were the only groups of plants under Series ІН that were rayed for eight weeks, the others being discontinued at the end of four weeks. A com- parison of these plants at the end of four weeks and eight weeks with similar plants rayed at 100 inches (F) will be seen in pl. 22, figs. 2 and 3. The set of plants having nine to ten leaves was found to be a little more resistant. At the end of one week the oldest leaves began to show tiny brown spots scattered over their surfaces. Soon after they began to dry up. At the end of three weeks all the older leaves were dead and the new ones smaller but not nearly as small as the new ones in the group having two leaves at the beginning. The leaves were very brittle and even the youngest very curly and brownish (pl. 22, fig. 1). At the end of eight weeks the rayed plants had an average of 18.32 leaves per plant and the controls 25.8 leaves (table 1). TABLE I SHOWING RATE OF GROWTH IN LACTUCA. FIGURES INDICATE THE AVER- AGE NUMBER OF LEAVES PER PLANT Series number Date ТН та ЇН та Ргев Lost Pres Lost Pres. Lost Pres. Lost Mar. 1 2; 0 200 00 0 0.00 0 аг. 8 3. 0 3.40 0 ‚60 2.00 1.60 1.30 Мат. 15 4.60 . 60 60 0 .40 4.50 2.20 2.60 Мат. 22 4.00 2.801 0.2 0 20 4.40 1.30 4.10 Маг. 29 4.30 3.10 7.00 | 1.50 60 2.10 4.30 1.33 рг. 5 5.10 .30 .90 10 2.40 8.10 1.00 Арг. 12 5.00 1.10 | 10 00 1.20 .66 5.92 | 21.20 8.90 Арг, 19 4.80 1.20 | 10.20 | 2.40 .38 4.00 | 25.80 | 3.90 Арг. 26 4.25 5.00 | 18.00 | 1.00 Net total 2.25 13.60 | 11.00 | 7.00 8.38 | 24.73 | 15.80 | 18.13 In Bryophyllum the first evidence of raying appeared the sixth day in the form of a glossy upper surface. At the end of two weeks the new leaves were very abnormal in form, the halves rolling upward from the midrib but the leaf itself not curling. Three sets of Phaseolus seedlings were used with six plants in each. One set had both cotyledons intact, another had one 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 183 cotyledon removed, and the third had both cotyledons removed. The object was to determine if the removal of stored food had any influence on the effect of raying. In all cases growth was re- tarded and burning resulted. The leaves became very blistered and abnormal in shape. The difference between rayed and control plants with both cotyledons removed was very great but the difference with one cotyledon removed was about the same as where both cotyledons were intact (table v). Series Г Е (rayed at 100 winches from the light without a screen).—The results with Cucumis here were in general the same, though never as marked for the same amount of raying, as in Series ТН. The appearance of injury was retarded several days, being noted first on the fifteenth day when the plants re- ceived an exposure of 74% minutes. For comparison of the size of rayed and control plants see table 1 and pl. 21, figs. 1 and 2. At the end of four weeks the average dry weight of rayed plants was 0.616 grams and the control plants 1.29 grams. At the end of eight weeks the leaves were as rolled as in Series I H. TABLE II SHOWING RATE OF GROWTH IN CUCUMIS. FIGURES EXPRESS AVERAGES PER PLANT Series number IF ще: Date Leaves Leaves Ht. in Leaves Leaves Ht. in pres. lost cm. pres. lost ст. Feb. 1 2.00 0 0 2.00 0 0 Ееђ. 8 6.00 0 7.35 5.25 6 ЕеЬ. 15 7.00 „га ‚5 ү 1.25 Ееђ. 22 7.50 20 11.60 8.00 11,25 Feb. 29 9.00 30 13.80 10.00 28 14.00 Net total 7.00 1.25 13.80 8.00 1:78 14.00 Тће experiment using Гротоеа plants was continued for eight weeks. А slight browning of the veins was noticed at the end of the eighteenth day when the plants received an exposure of nine minutes. At the end of four weeks the usual blistering appeared as can be seen in pl. 21, fig. 6. About as many leaves were added as to the control plants but they never attained as large a size. (Vou. 15 184 ANNALS OF THE MISSOURI BOTANICAL GARDEN Nicotiana behaved very much the same here as in Series I H except that the effects were a little later in appearing. The same four varieties of Coleus Blumei were used as in the pre- ceding series and were again divided into two groups. The ef- fect on both groups was somewhat less marked and much retarded, particularly as far as shedding leaves was concerned. The more resistant group shed no leaves until the fifth week, and at the end of seven weeks only a few had been shed (pl. 23, figs. 1-4). When these plants were put under normal greenhouse conditions at the end of eight weeks raying, they began to show normal growth and color after ten days, but at the end of four weeks they were still far behind the control plants (pl. 22, fig. 5). Two sets of Raphanus were used, one with two leaves and the other with four to five leaves. Here there seemed to be very little difference in the effect produced whether the plant was just above ground or had several leaves. The effect of raying did not appear until the eighteenth day, but at the end of four weeks it was quite marked (pl. 21, fig. 5). At the end of eight weeks there was a noticeable difference in the number of leaves, the rayed plants averaging 6.2 and the control 8.42 leaves per plant (table пл (a), and pl. 21, fig. 4). The same two sets of Lactuca plants were again used. The same general results were found for the set having two leaves, but in the set having nine leaves the rayed plants produced as well as lost more leaves than the controls though they were never as large. A comparison of the effects produced here with those in Series I H can be well seen in pl. 22, figs. 1, 2, and 3 and table пт (b). In addition to the above two sets of plants two more sets were used, one set consisting of two-leaved lettuce plants of a red variety and the other of old lettuce plants with fifteen leaves. Red lettuce was rayed to see if the color would disappear as it had done in Coleus. However, after raying for eight weeks the red color was still evident though partly masked by the brownish effect of raying. The old Lactuca plants were used in order to determine the effect of ultra-violet light on bud and flower formation. The same retarding effect was found though less with these older plants. At the end of eight weeks both rayed and control plants were 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 185 budded. The flower stalks of the control plants were greater in diameter and seemed to branch more at the top (pl. 26, fig.1). Bryophyllum rayed under these conditions showed no curling of the leaves, due no doubt to their thickness. However, the leaves again rolled upward from the midrib. At the end of two weeks they had shiny brownish surfaces similar to those found in many of the other plants, and often parts of the new leaves formed were undeveloped. For a comparison of the rate of growth in rayed and control plants see table ш (с) and pl. 26, fig. 5. The ten Zea Mays seedlings were found to be as resistant to ultra-violet light as any of the plants used, showing the first evidence of any harmful effect the twenty-fourth day when they received a twelve-minute exposure. Even then the effect was slight, taking the form of a slight rolling upward of the edges of the leaves. When the rate of growth was compared, the rayed TABLE III SHOWING RATE OF GROWTH IN SERIESI F AND G. FIGURES INDICATE THE AVERAGES PER PLANT (a) Raphanus (b) Lactuca Date IF IG IF IG IF IG Pres.| Lost | Pres.| Lost | Pres.| Lost | Pres.| Lost | Pres.| Lost | Pres.| Lost Mar. 1 2.00 0 .00 0 | 2.00 0 | 2.00 0 | 9. 019.0 0 Mar. 8 4.00 0 | 3.91 0 | 3.75 0018 7 0 112.0 | 1.5010.4 | 2.2 Мат. 15 | 5.63 0 | 5.63 0 | 4.28 014.2 0 {12.9 | 2.7010.6 | 2.1 Mar. 22 | 6.90) 0.09| 6.16| 0.031 5.21 1.7 | 7.0 0 112.6 3.6510.75 2.3 Маг. 29 | 6.90) 1.00) 6.53] 0.03] 6.07! 3.2 | 7.4 | 1.7 110.35 5.80 9.6 | 4.2 Арг. 5 6.60} 1.60] 6.64| 1.06 6.85) 1.4 | 9.2 | 0.7 112.6 | 1.80112.4 | 1.5 Арг. 12 | 7.20] 1.00] 7.71 0.10 8.80] 1.7 11.8 | 0.4 |15.5 | 2.50115.5 Ж Арг. 19 | 6.20] 1.60] 8.42) 0.18 8.77 3.3 |12.6 | 2.2 119.0 | 3.66|18.4 | 3.5 Арг. 25 11:55] 1.6 116.4 | 1.8 122.1 1 24020.68 1 1.5 Net total| 4.20| 5.29| 6.42| 1.40| 9.5512.9 114.4 | 6.8 13.1 123.01 11.6 16.02 (c) Bryophyllum (d) Zea Mays Date IN IG IF IG Lvs. Ht. in Lvs. Ht. in Lvs. Ht. in Lvs. Ht. in pres. cm pres. cm. pres. cm pres cm. Jan. 24 12.59 13.14 13:11 14.35 ДЕ реа 35.24 Feb. 1 13.41 13.85 14.28 15,41 3,20 ады 3.50 3.62 Feb. 8 14.85 15.14 | 15.71 17.35 3.75 5,2 4.00 5.37 Feb. 15 15.14 16.21 16.85 18.57 4.50 6.37 5.00 8.00 Feb. 22 17.00 17.04 | 17.42 | 19.07 5.00 8.43 6.25 9.94 Feb. 29 6.00 10.60 6.90 14.20 Net total 4.41 3.90 89.11 4.72 6.00 10.60 6.90 14.20 [Vor. 15 186 ANNALS OF THE MISSOURI BOTANICAL GARDEN plants showed a noticeable retardation (table пт (4) and pl. 26, fig. 4). SERIES П Plants in this series were under exactly the same conditions as those in Series I except that here a screen of vita glass was used. The series was again divided into three parts, group A, rayed at 50 inches from the light, group B, rayed at 100 inches, and group E, which was the control for both this series and Series ПТ. АП plants rayed were treated daily for seven weeks. Series II A (rayed at 50 inches from the light with a screen ој vita glass).—Plants rayed under these conditions responded very differently. In ten young Cucumis seedlings exposed to ultra-violet leaves were produced a little more rapidly than in the control plants (Е) but at the end of seven weeks the control plants had slightly more leaves than the rayed ones. The size and color seemed to be about the same for both. The stems elongated almost equally for the first three weeks and then increased very rapidly in the rayed plants, so that at the end of seven weeks they averaged eight to nine centimeters longer and were noticeably greater in diameter than the control plants (table ту (a) and pl. 24, fig. 1). Flowers appeared on the rayed plants two days earlier than on the control plants. At the end of seven weeks the rayed plants averaged 2.3 flowers and the control plants only 1.2 flowers per plant. Ten cuttings of Гротоеа of as near the same size as possible were rayed. Even at the end of seven weeks there was very little difference in height between them and the controls, though the average number of leaves added was much greater in the rayed plants (table ту (b) and pl. 24, fig. 2). The leaves of both rayed and control plants were of about the same size and color. Observations on fifteen Nicotiana plants point toward a retardation of growth, though there was no evidence of any burning. The leaves were about the same in number, size, and color. The stem, however, was taller and smaller in diameter in the control plants. No difference was noticed in the time of 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 187 flowering. In general, the rayed plants gave the appearance of being more stocky (table rv (c)). Ten young Zea Mays seedlings were used. At first the control plants grew taller, measuring from the base to the highest node. During the last few days, however, the rayed plants grew very rapidly and surpassed the controls. The rayed stalks were also larger in diameter. The leaves on the rayed plants not only cutnumbered those on the control plants, but were from 1.5 to 2.0 cm. wider in the middle, those on the controls averaging 4.0 cm. in width (table ту (d) and pl. 24, fig. 3). From the very beginning the twenty rayed plants of Lactuca produced slightly more leaves than the controls. The leaves of both were the same in color, texture, and size (table ту (e) and pl. 24, fig. 4). The size of the leaves on the ten rayed Raphanus seedlings equalled those on the controls, but there were slightly more leaves оп the latter (table ту (1) and pl. 25, fig. 1). Coleus Blume vars. “Spotted Gem" and Verschaffeltii have been found to be the varieties most sensitive to ultra-violet light, and six cuttings of each as near the same size as possible were used. Both varieties showed the same characteristics. From the very beginning the rayed plants showed greater growth both as to number and size of leaves, and as to height and diameter of stem (table ту (h and i), and pl. 25, figs. 2 and 3). The red color did not seem to be affected as it was in Series I. Eight cuttings of Bryophyllum were rayed. At first the controls grew taller, with a greater number of leaves, but during the last few weeks the rayed plants much surpassed the controls (table ту (g) and pl. 25, fig. 4). Three sets of Phaseolus seedlings of six each were used as in Series 1. Those with both cotyledons intact and with one cotyledon removed were taller and had more leaves than the corresponding control plants. Those with both cotyledons removed had slightly more leaves than the corresponding control plants, but were shorter (table v). Series 11 В (rayed at 100 inches from the light using a screen of vita glass).—The same number of Cucumis plants was used as in Series ГА. From the very first the number of leaves on the Мог. 15 188 ANNALS OF THE MISSOURI BOTANICAL GARDEN rayed plants exceeded those on the controls, but the size of the leaves was about the same in both. The rayed leaves in this group seemed a little deeper green than did the controls. АП through the experiment the stems of the rayed plants increased in both length and thickness more rapidly than those of the control plants, and at the end of seven weeks were a little greater in length than the stems of the plants in Series II A (table rv (a) and pl. 24, fig. 1). About the same number of leaves was present on the ten Ipomoea cuttings in this series as in Series П A, which was much greater than in control plants. The stem, however, was greater in length here than in either Series ПА or the control Е (table ту (b) and pl. 24, fig. 2). In the fifteen Nicotiana plants rayed little difference was found from Series II A either in number or color of leaves or in length and thickness of stem. The control plants had about the same number of leaves but were taller (table ту (с)). In this group the ten Zea Mays seedlings showed a slight increase in number of leaves over those in Series ПА and a greater increase over the control plants in E. In average growth in height and diameter of the stalk this group exceeded both the control plants in E and the rayed plants in A (table rv (d) and pl. 24, fig. 3). Here, as in Series ПА, twenty Lactuca plants were rayed. All through the experiment there was a slight increase in number of leaves over that in the control plants though the size of the leaves was a little greater in the control plants (table rv (e), and pl. 24, fig. 4). The ten rayed Raphanus seedlings showed leaves equalling those of the control plants in size but fewer in number. The seedlings in this group were almost identical with those in Series II A (table rv (f), and pl. 25, fig. 1). The same number of Coleus cuttings was used as in Series П A. As to number of leaves produced the plants in this group about equalled those in Series II A, but much surpassed the controls in Е. Here the plants exceeded in height both those in Series П А and the controls E (table rv (h and i) and pl. 25, figs. 2 and 3). The Bryophyllum cuttings in this group produced about the 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 189 same number of leaves as the controls but increased a little more in height than did the controls. Plants in Series II A surpassed this group both in number of leaves and in height (table ту (с) and pl. 25, fig. 4). SERIES Ш The same conditions were present in this series except that instead of vita glass a screen of quartz-lite glass was used. This series also was divided into three parts, group C, rayed at 50 inches from the light, group D, rayed at 100 inches, and group E, again the control. The same number of plants were used in each case as in Series II. Series III C (plants rayed at 50 inches from the light using a screen of quartz-lite glass) —The number of leaves produced оп rayed Cucumis seedlings was a little more than in Series II A, but a little less than in Series IIB. As to growth in height both Series ПА and В surpassed this group by about three centimeters. However, this group surpassed the control by more than five centimeters (table rv (a) and pl. 24, fig. 1). The /ротоеа cuttings in this group produced more leaves than in either A or В of Series II, though the growth in height was a little less here than in those groups. This group showed better growth than the control in all respects (table rv (b) and pl. 24, fig. 2). There was little difference between the growth of the Nicotiana plants here and in groups A and B of Series II. The control plants surpassed all three mentioned groups in height, but about equalled the other groups in number of leaves (table 1v (c)). The Zea Mays seedlings here were almost identical with those in Series II B as to number and size of leaves and as to height of plant. It would be hard to determine from external observation which of these two groups produced the better growth (table rv (d) and pl. 24, fig. 3). The Lactuca plants in this group showed more leaves than either the control plants or the plants in Series II A and B. However, the leaves in this group of plants were a little smaller than in the control plants (table rv (e) and pl. 24, fig. 4). The average number of leaves present in Raphanus seedlings in this group was slightly less than in the controls, but otherwise the plants were identical (table rv (f) and pl. 25, fig. 1). [Vor. 15 190 ANNALS OF THE MISSOURI BOTANICAL GARDEN Both varieties of Coleus again showed similar results. The cuttings in this group showed more growth in height and number of leaves than did the controls but not as much as in either A or B of Series II (table rv (h and 1) and pl. 25, figs. 2 and 3). The Bryophyllum cuttings in this group surpassed the control plants and those in Series П В both in number of leaves and in height. However, the growth in this group did not equal that in Series II A (table rv (g) and pl. 25, fig. 4). As in Series II A, three sets of Phaseolus seedlings were used. Those with both cotyledons intact and with one cotyledon removed were a little taller and had slightly more leaves than the control plants, but were not quite as tall nor did they possess quite as many leaves as those in Series ПА. The plants with both cotyledons removed showed less growth in height than the corresponding controls, but a little more than those in Series П А. The number of leaves present differed very little (table v). Series III D (plants rayed 100 inches from the light using a screen of quartz-lite glass).—Cucumis seedlings in this group differed very little from those in group C and thus were several centimeters greater in length than the controls (table rv (a) and pl. 24, fig. 1). Ipomoea plants showed greater growth in height in this series than in Series ПТС, with about the same number of leaves present in each series (table ту (b)). The Nicotiana plants closely resembled those in Series II A and В, being stocky while the controls were much taller (table ту (c)). Seedlings of Zea Mays in this group were not quite as tall as in the other groups mentioned, but nevertheless surpassed the control plants. As to number of leaves present this group about equalled the other groups. All the rayed groups in Series II and III, as previously mentioned, possessed leaves from one and a half to two centimeters wider in the middle than the control leaves (table ту (4) and pl. 24, fig. 3). Lactuca plants in this group gave a slightly smaller count of leaves than in Series III C, but more than in Series II A and B and also more than the control plants. Little difference was noticed between the size of the leaves here and in the control plants (table ту (e), and pl. 24, fig. 4). 1928] ELTINGE—EFFECT ОЕ ULTRA-VIOLET RADIATION 191 Raphanus plants showed slightly fewer leaves than in Series III C and thus fewer than the control plants (table rv (f) and pl. 25, fig. 1). Both varieties of Coleus plants in this group surpassed the control both in number of leaves and growth in height, but showed fewer leaves and less growth in height than in Series II A and В and Series III С (table ту (h and i) and pl. 25, figs. 2 and 3). Cuttings of Bryophyllum showed fewer leaves than in Series ПА or Series ПТС and about the same number as the control plants and Series II B. However, this group showed almost as great growth in height as in Series III C, which surpassed Series II A and B and also the controls (table rv (g)). TABLE IV SHOWING THE RATE OF GROWTH OF PLANTS IN SERIES II АМР ПТ. FIGURES REPRESENT AVERAGE N {BER ВОР LEAVES PER PLANT; AND HEIGHT (a) Cucumis Series II Series III Control Date A B С р Е Lvs Ht Lvs Ht Lvs. | Ht. | Lvs Ht Lvs. | Ht Oct. 24 2.00 2.00 2.00 2.00 Oct. 31 3.50 | 7.45 | 3.80 6 3.80 | 6.35 | 3.60 | 7.75 | 3.00 6.8 ov. 7 5.00 | 8.90 | 5.00 [10.00 | 4.90 | 8.30 | 5.00 | 9.85 | 4.80 7.9 Nov. 14 5.60 | 9.85 | 6.00 [11.20 | 5.87 .50 | 6.00 110.60 VA 9.5 Nov. 21 7.10 13.00 | 6.70 |14.00 | 6.77 |11.33 | 6.80 |13.05 | 5.90 | 11.1 Nov. 28 7.20 |16.90 | 7.00 119.20 | 6.77 |14.83 | 6.80 |17.15 | 6.77 | 13.0 ec. 7.10 122.70 | 8.11 124.72 | 7.66 120.00 | 6.80 122.25 .44 | 17.6 Dec. 12 7.30 129.05 | 8.22 29.77 | 7.77 |26.04 | 7.10 127.90 | 7.65 | 20.5 Net total 5.30 |29.05 | 6.22 29.77 5.77 |26.04 | 5.10 27.90 | 5.65 | 20.5 (b) Ipomoea Series II Series III Control Date A B C D E Lvs Ht Lvs Ht Lvs. | Ht Lvs Ht Lvs Ht Oct. 24 ERAZ 11.251 7.25 112.8 8.44 | 8 6 | 5.87 | 5.75 Oct. 31 14.62 111.5 9.78 13.81 [11.8 9.64 11 11.87 | 6.75 | 6.25 оу. 7 16.75 12.62 [11.75 14.81 113.62 [10.59 113.00 12.2 8 6.94 Nov. 14 118.25 3 114 .21.116.00-111.37. 16.25 112.81 110.31 172 97 Nov. 21 [20.70 114.50 [18.00 [16.57 [19.87 |12.75 |17.62 14.06 [10.75 | 8.43 Nov. 28 (24.00 [15.18 [22.00 [17.42 [24.56 |15.21 122.00 115.31 112.75 | 9.81 D 5 26.75 |15.68 |25.71 119.57 |27.42 |16.50 126 15.42 |14.42 |10.35 ес. 12 |28.80 116.50 [27.33 120.66 130.14 117.85 (81.51 118.00 14.50 10.50 Nu 17.68 | 5.25 |20.08 | 7.84 |21.02 | 9.41 |23.26 | 9.44 4. ГУ ог, 15 192 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN (c) Nicotiana Series IT Series ITI Control Date A B С р Е Lvs. | Ht. | Lvs. | Ht. | Lvs. | Ht. | Lvs Ht. | Lvs Ht. Oct. 24 4.46 4.10 4.10 4.46 4.46 Oct. 31 6.40 6.40 6.40 6.60 6.13 Nov. 7 7.93 .58 8.06 8.28 7.53 Nov. 14 |9.80 10.07 9.80 9.07 9.21 Nov. 21 9.50 | 5.00 | 9.71 9 110.2 5.03 | 9.70 | 4.69 | 9.57 | 5.85 Nov. 28 [10.41 | 6.12 Ч 2 6.66 Уа E 7.05 | 9.80 | 6.65 |10.25 | 8.08 ec. 5 2.20 | 9.25 9.45 8.95 |11.40 | 9.05 |12.55 |15.00 Dec.12 113.20 13.05 + % 13.45 T$ 79 13.25 [18.20 [18.80 112.88 |19.05 № total | 8.74 113.05 | 9.85 {13.45 | 8. 60 13.25 8. 74 13.30 | 8.42 119.05 (4) Zea Mays Series П Series III Control Date A B С р Е Lys. | Ht. | Lvs. | Ht. | Lvs. | Ht. | Lvs Ht Lys Ht Oct. 24 | | | Just above ground | | Oct. 31 3.50 | 4.62 | 3.62 | 5.18 | 3.38 | 5.19 | 3.50 | 4.62 | 2.88 | 4.77 ov. 4.38 | 7.31 | 4.88 | 7.79 | 4.50 | 7.87 | 4.62 | 7.56 | 3.80 | 7.55 Nov. 14 5.12 | 7.93 | 5.62 | 9.43 | 5.37 | 9.31 | 5.25 .37 | 5.20 | 8.60 Nov. 21 | 9.4 6.75 112.00 | 5.66 110.94 | 6.25 110.68 | 5.60 [10.20 Nov. 28 6.75 112.93 | 7.85 16.25 | 7.11 114.61 | 7.37 114.31 | 6.40 14.10 Dec. 5 ‚00 115.56 | 8.14 118.57 | 8.00 117.22 | 8.33 |16.81 | 7.00 |15. Dec. 12 | 9.00 [18.75 | 9.71 (20.00 | 9.22 [20.00 | 9.28 [18.42 | 8.00 [17.3 (e) Lactuca (f) Raphanus Series П Series ПТ | Control| Series II Series III | Control Date A B а р Е А В С р Е Lvs. | Lvs. | Lvs. | Lvs Lvs Lvs. | Lvs. | Lvs. | Lvs. | Lvs. Oct. 24 .25 | 8.1 2 8.001 8.1 2.00 | 2.00 | 2.0 0 2.00 Oct. 31 13.35 |12.35 113.15 113.15| 12.65 | 2.80 | 2.90 | 2.3 3.301 2.90 ov. 7 6.90 116.45 116.95 116.45] 15.70 | 4.20 .40 | 4.4 8 4.20 Nov. 14 120.80 |20.45 [21.65 |20.35 20.25 .70 ›.00 | 5.4 5.70| 5.80 Nov. 21 122.60 (23.25 |25.20 |21.60| 20.35 .80 .60 | 5.90 701 5.60 Nov.28 125.61 |25.27 |25.05 |24.52| 23.11 .80 .60 | 4.9 5.10 ot Dec. 5 7.07 |27.78 |26.68 129.06| 25.85 ).55 ›.60 | 6.1 1 .00 Dec. 12 128.38 |27.57 (82.18 (80.52| 26.90 ХА 7.1011 6.6 6.40| 7.60 Net total (20.13 [19.47 |23.98 22.52| 18 18. 80 5.22 5.10 | 4.6 40} 5.60 1923] (в) Bryophyllum ELTINGE—EFFECT ОЕ ULTRA-VIOLET RADIATION 193 Series П Series ПІ Control Date B С р Е Ht Lvs. Ht Lvs: | НЕ има: 1 Ht Oct. 31 7.46 | 8. 6.61 | 8.50 | 5.34 | 6.75 | 5.25 ov. 7 7.62 11.00 7.05 110. 87 | 8.00 | 7.18 Nov. 14 8.81 111.25 8.16 110.75 | 6.63 ОБ | 7.70 Nov. 21 187 11.26 XT ЧИДЕ 00 110.25 | 7.94 Nov. 28 ТО To 2:12 11.44 12.50 10.12 10.75 | 8.25 ес. 5 13.85 13.55 110 11.50 12.75 111.25 11.25 | 9.25 Пес. 12 37 14.94 14.12 ! 13.61 114. 13.37 112.25 110.00 Net total 0 | 7.48 | 5.46 | 6.71 7.00 | 5.50 | 8.30 .90 | 3.75 (h) Coleus Blumei var. Verschaffeltii Series II Series Ш Control Date C D E Ht, |-Lvs. Ht Lvs Ht Lvs. | Ht Oct. 24 3.75 | 9.16 83 | 3.83 | 9.50 | 4.00 | 6.83 | 3.2 Oct. 31 4.58 |12.00 5014838 11.33 | 5.0 .88 | 8.0 Nov. 7 5.66 |17.6 00 | 6.25 117.16 | 6.25 11.00 | 4.6 Nov. 14 6.46 |25.66 7.41 126.33 | 7.75 14.00 | 5.3 Коу. 21 8.33 |34.00 0 | 9.40 |29.33 | 9.50 115.50 | 6.1 Nov. 28 9.50 (43.66 0 |10.70 (37.66 110.5 ЯЗ 1 7.5 ec. 5 10.66 |51.16 .60 |12.10 143.83 |12.66 |25.83 |10.0 Dec. 12 14.40 (69.80 .60 114.30 158.50 114.26 35.16 [10.7 Net total 10.65 160.14 .77 10.47 149.00 10.26 (28.33 | 7.00. (i) Coleus Blumei var. “Spotted Gem" Series IT Series III Control Date С р Е Ht Lvs Ht Lvs Ht. | Lvs. | Ht Oct. 24 3.33 | 9.6 7.00 | 1.66 00 | 2.00 | 6.00 | 2.00 Oct. 31 6.33 112.00 ret 2.08 1245 2. 8.66 | 2.16 Nov. 7 6.50 |20.30 9.3 2.75 111.00 | 3.06 |10.66 | 3.00 Nov. 14 7.58 |28.33 14.00 | 3.33 116.66 | 3.33 |12.66 | 3.16 Nov. 21 9.33 |41.00 20.0 4.50 |19.00 | 4.75 |12.66 | 3.83 Nov. 28 11.16 151.30 [13.33 (26.60 | 5.16 [20.00 | 6.16 [15.80 | 4.83 ec. 5 13.16 |64. ) 5.66 |22 66 | 6.83 |18.00 | 6.33 Гес. 12 178 15.66 (84.00 [18.00 (36.60 | 7.16 137.60 | 8.10 [25.66 | 7.10 Net total 12.33 174.34 |14.50 29.60 | 5.50 (31.60 | 6.1 19.66 | 5.10 [Vor. 15 194 ANNALS OF THE MISSOURI BOTANICAL GARDEN TABLE V (a) оно RATE OF GROWTH IN PHASEOLUS SEEDLINGS WITH BOTH OTY HERG nea PRESENT. FIGURES REPRESENT AVERAGE NUMBER AVES PER PLANT AND HEIGHT IN CENTIMETERS Series I H Series II A Series ПТС Control Е Date Lys. Ht. Lvs. Ht. Lvs. Ht. Lvs. Ht. Oct. 28 2.00 5.25 | 2.00 4.37 2.00 4.66 | 2.00 .21 Nov. 5 8.75 | 10.87 3.33 | 10.60 3.50 | 12.00 8.15 | 10.75 Моу. 12 5.00 | 12.75 4.33 | 13.60 5.00 | 13.83 4.50 | 13.37 Nov. 19 5.50 | 15.00 5.00 | 17.20 5.10 | 17.91 4.80 | 15.71 Nov. 26 5.50 | 15.80 6.00 | 22.80 5.30 | 22.91 5.00 | 21.08 Net total 3.50 9.55 4.00 | 18.43 | 3.30 | 18.25 8.00 1 16.87 Av. no. flowers 0 15 (b SHOWING RATE OF GROWTH IN PHASEOLUS SEEDLINGS WITH ONE COTYLEDON REMOVED Series I H Series ПА Series Ш С Control Е Date Lvs. Ht. Lvs Ht. Lvs. Ht. Lvs Ht. Oct. 28 2.00 . 62 2.00 4. 2.00 4.66 2.00 .50 Nov. 5 3.25 | 10.37 3.00 | 10.40 3.10 9.16 3.30 .20 Nov. 12 4.00 | 12.37 4.20 | 13.00 5.10 | 12.70 4.00 | 11.20 Nov. 19 5.00 | 14.56 4.90 | 16.40 5.20 | 18.80 4.50 | 13.50 оу. 5.00 | 15.00 5.60 | 20.70 5.20 | 20.58 5.00 | 14.75 № tota 3.00 .38 8.60 | 16.54 3.2 15.92 3.00 | 11.25 Av. no. flowers ) я .8 1.4 (с) SHOWING RATE OF GROWTH IN PHASEOLUS SEEDLINGS WITH BOTH COTYLEDONS REMOVED Series I H Series ПА Series III C Control E Date Lvs. Ht. Lvs. Ht. Lvs. Hi. Lys. Ht. Oct. 28 2.00 5.25 2.00 3.33 2.00 5.33 2.00 4.50 Nov. 5 2.25 8.00 2.50 7.25 2.70 7.46 2.50 | 8.90 Коу. 12 3.30 9.87 3.60 4.80 | 11.00 4.20:1:12:82 Nov. 19 4.80 | 11.50 4.20 | 11.41 4.80 | 13.40 4.50 | 14.36 Nov. =. 4.30 | 11.75 5.10 | 15.00 4.80 | 18.50 4.70 | 18.20 Net tot 2.30 6.50 8.10 | 11.67 2.80 | 13.17 2.70 | 13.70 Ау. по. y ТМВ 0 : : 14 According to Beeskow (728) and Delf and Ritson (Delf, Ritson, and Westbrook ’27) a daily exposure to ultra-violet of as long as thirty seconds produces no harmful effects in Soy beans and Tri- folium and might cause a slight increase in growth. Thus several experiments were undertaken to see if this might not be true of other plants. Fifteen young Nicotiana plants were rayed at 100 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 195 inches from the light without a screen for thirty seconds each day for a period of four weeks. At the end of that time the rayed plants showed a noticeable increase in growth over the control plants (table уг). The same experiment was tried with three varieties of Coleus with the same results. Here the increase was not only in number of leaves, but also in height (table уг and pl. 26, fig. 2). Very young lettuce plants were also rayed with the same results. Next, the exposure of one minute each day at 100 inches from the light was tried on the same three varieties of Coleus, with absolutely no change in color. There was, however, a slight retardation in the rate of growth at the end of two weeks, but even at the end of four weeks it was not very noticeable. TABLE VI SHOWING RATE OF GROWTH IN PLANTS RAYED 30 SECONDS EACH DAY AT 100 INCHES FROM THE LIGHT, USING NO SCREEN (a) Nicotiana (b) Coleus Blumei var. Verschaffeltii Date Rayed Control Rayed Control Lvs. pres. | Lvs. pres. | Lvs. pres. |Ht. іп em. | Lvs. pres. | Ht. in em. Jan. 24 4,26 3.93 12.25 7.50 10.60 7.83 Feb. 1 5.60 5.26 15.50 11.30 8.00 eb. 8 7.10 7.18 28.00 10.00 16.60 Feb. 15 8.46 7.66 28.75 12.12 21.00 10.00 Кер. 22 9.93 8.66 85.45 12.75 29.00 11.30 Total 5.67 4.73 23.20 5.25 18.40 3.34 (с) Coleus Blumei var. “Defiance” (d) Coleus Те „Уат. “Spotted Date Rayed Control Rayed Control Lvs. Ht. in | Lvs. | Ht.in| Lvs. | Ht.in | Lvs. | Ht. in pres em. pres em. pres cm pres em. Jan. 24 11.00 8.37 7.00 4.00 | 13.50 8.62 | 15.75 | 8.87 Feb. 1 14.50 7.50 4.52 | 16.25 7 | 18.25 | 9.25 еђ. 8 19.25. | 11.87 8.75 6.25 | 32.75 | 10.62 | 33.50 | 9.75 Feb. 15 24.75 | 15.00 25 8.5 47.00 | 12.37 | 44.75 | 11.25 Feb. 22 26.5 16.50 | 13.7 10.87 | 49.7 12.75 | 50.50 | 12.00 Total 15.50 13 5 6.3 36.25 4.13 | 34.75 1 ГУ он, 15 196 ANNALS OF THE MISSOURI BOTANICAL GARDEN ANATOMICAL (ALL MEASUREMENTS WERE MADE WITH AN EYEPIECE MICROMETER) Leaves.—A cross-section of a Cucumis leaf rayed under the conditions present in Series I F was measured and found to be a little thinner than a corresponding section from a non-rayed leaf. When the structure of the two sections was compared, it was observed that the raved section was lacking in upper epidermis, with only the collapsed walls remaining as a false euticle, which no doubt gave the glossy surface to the leaf. Also the protoplasm in most of the palisade cells had drawn away slightly from the ends of the cells nearest the epidermis. The chloroplastids in the rayed section were found to be more numerous in the ends of the palisade cells nearest the spongy tissue. Another difference was the presence of fewer air-spaces in the rayed section (pl. 28, figs. 4 and 5 and table уп (b)). This corresponds well with the results found by Bailey (’94), Kluyver (711), and Westbrook (Delf, Ritson, and Westbrook, 727). When a cross-section of a Cucumis leaf, rayed as in Series II A, was measured it was found to be a little thicker than a similar section from a non-rayed plant. А leaf from Series II B was found to be still thicker. It will be remembered that this was also the group where greatest growth in height was found. A leaf from Series III C was thicker than Series II A but thinner than Series ПВ. A leaf from Series III D was slightly thicker than the control though not as thick as a leaf from Series II A. The control leaf showed very long palisade cells with many air- spaces between them and also among the cells of the spongy tissue. In Series II B where the thickest leaf was found, there were larger air-spaces than in the control leaf. The palisade cells in all rayed leaves in Series II and III were a little shorter than in the control leaves. The thinnest rayed leaf (Series III D) showed fewer air-spaces than the control and instead smaller and more numerous cells. The number and position of the plastids was about the same in all leaves (pl. 29, figs. 1-3, and table уп The cross-section of a rayed leaf of Ipomoea from Series I H was found to be much thinner than an unrayed leaf. Неге, as in Cucumis, the epidermis had collapsed, forming a heavy cuticle. 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 197 Occasionally, however, an epidermal cell remained intact. The protoplasm of a few of the palisade cells had drawn away from the end nearer the epidermis, and the cells themselves were shorter. A section from an Ipomoea leaf in Series I F proved to be of Ше same thickness as the one from Series ІН. Here, however, the epidermis was not collapsed but thinner, each cell being absolutely distinct. The palisade cells were longer than in Series I H but not as long as the controls. There were fewer air-spaces here than in either the control leaf or the leaf in Series IH. This is probably due to the fact that Series I F was rayed for a period of eight weeks and Series I H for only four weeks (table уп (d)). When rayed Ipomoea leaves from Series II A were examined, they were found to be thinner than control leaves. Leaves from Series II B were slightly thinner than those in II А. The palisade cells in both the above-mentioned groups were thinner than palisade cells in control leaves, as were also the upper epidermal cells. Rayed leaves in Series III C about equalled the control leaves in thickness and length of palisade cells, though the upper epidermis here was still thinner than in the control leaves. Leaves from Series III D were by far the thickest in any of the series. Here the palisade cells and epidermal cells were also longer than in any of the other groups. Air-spaces were found to be much greater and more numerous than in sections of control leaves. Chloroplastids seemed to be more numerous here also (table уп (p), and pl. 29, figs. 4 and 5). The cross-section of а rayed Nicotiana leaf from Series I H was again found thinner than an unrayed leaf. The same collapsing of the epidermis and shrinking of the protoplasm in the palisade cells were also found (pl. 27, fig. 7, and table уп (e)). Rayed Nicotiana leaves from Series II A and B and III C were about the same thickness as those of non-rayed plants. The palisade cells from Series II A and B were a little longer than corresponding cells in a control leaf. А Nicotiana leaf from Series III D proved to be much thicker than one from any of the : other groups mentioned. Also its palisade cells were longer, and it contained many more air-spaces. However, the upper epidermis [Vor. 15 198 ANNALS OF THE MISSOURI BOTANICAL GARDEN was thinner here than in a non-rayed leaf (pl. 27, figs. 5 and 6, and table vir (0)). It will be remembered that this group, while it did not show greatest growth in height, was the most stocky in appearance and produced flowers at the same time as did the control plants showing greatest growth in height. When cross-sections of Zea Mays leaves were examined it was found that all rayed leaves in Series II and III were much thicker than non-rayed ones, the thickest being present in Series III C, which was rayed for seven weeks at 50 inches from the light using а screen of quartz-lite glass. These leaves also showed the thinnest epidermis and the best-developed vascular bundles. Leaves in Series II A and B were next in thickness, and both had well-developed bundles. Leaves in Series III D were the nearest like those of the control plants, having very thick epidermis and less well-developed bundles. In general, there seemed to be a thicker cuticle present in rayed leaves than in control leaves (pl. 32, figs. 1-6, and table уп (n)). Cross-sections of Coleus leaves in Series I H which were rayed at 50 inches without a screen showed the same collapsed epidermis and lack of air-spaces as were found in similarly treated leaves of other plants. However, here in addition there was a loss of red color. Not only did the color disappear from the epidermis when it collapsed, but also from the palisade cells, showing that the rays penetrate beyond the epidermis or else produce some substance which does. This corresponds well with the results found in animal tissue by Macht, Anderson and Bell (728) (pl. 31, figs. 4 and 5). Non-rayed Coleus leaves were found to be much thicker than those rayed. The thinnest leaves were found in Series II B, where there was also the greatest growth in height. According to increasing thickness the groups may be arranged as follows; Series II B, ПА, Ш С, III D, and last, the control E. The palisade cells were shorter in Series II В than in any of the other groups, even including the control plants. There was absolutely no decrease in red color in any of the rayed plants in Series П and ПТ. It appeared that in Coleus plants the growth in height was inversely proportional to both the thickness of the leaves and the number of air-spaces present (pl. 30, figs. 1-5, and table уп (1)). 1928] ELTINGE—EFFECT ОЕ ULTRA-VIOLET RADIATION 199 Lactuca leaves rayed as in Series I H showed the characteristic collapse: of at least part of the epidermis and the lack of air-spaces. In addition, there was no differentiation of palisade tissue. Leaves rayed as in Series I F for four weeks were similar to the controls except that they were a little thinner and developed palisade and epidermal cells that were a little shorter. The air- spaces were also fewer here than in control leaves. If their leaves were rayed for eight weeks instead of four, they were still thinner, had no well-defined palisade layer, and almost no air-spaces. In thickness they about equalled the leaves in Series I H When rayed Lactuca leaves from Series II and III were examined they were all found to be of about the same thickness and much thinner than corresponding control leaves. The palisade and epidermal cells were also shorter than those present in the control leaves. Cross-sections of rayed Raphanus leaves in Series I F were only very slightly thinner than those of corresponding control leaves. There was the same collapse of epidermis, forming a false cuticle as in other leaves mentioned. Тһе contents of the upper layer of palisade cells had disappeared, leaving them empty. Leaves in Series I H showed about the same injury as those in Series IF rayed for eight weeks. Кауеа leaves in Series II and III were all thinner than similar control leaves, those in Series III D being the thinnest. The palisade and epidermal cells, however, seemed to be longer than in the control leaves. This would indicate fewer air-spaces in the rayed leaves in Series II and III than in corresponding non-rayed leaves. Sections of Bryophyllum leaves rayed under the conditions of Series I H were also thinner than corresponding sections of non- rayed leaves. Тһе characteristic lack of upper epidermis was | observed and also a thicker under epidermis. Sections of leaves from this plant in Series ПА were much thinner than those of control leaves, and there was no collapse of upper epidermis. Leaves from Series II B were about the same thickness as control leaves, but the epidermis was slightly thinner. Leaves in Series III C were thinner than in Series II B but thicker than in Series ПА. Leaves from Series ПТО had longer upper epidermal (Vor. 15 200 ANNALS OF THE MISSOURI BOTANICAL GARDEN cells and were much thicker than any of the other leaves includ- ing the controls. Cross-sections of Phaseolus leaves in Series I F were thinner than sections from control leaves. The epidermis was destroyed in some places, but in others the epidermal cells still remained intact. The contents of the palisade cells were much shrunken and the cells shorter. The chloroplastids were also collected in the end of the cells nearest the spongy tissue. Sections of Phaseolus leaves from Series ПА were much thicker than control sections. They also had longer palisade and epidermal cells and more numerous air-spaces and chloroplastids. Phaseolus leaves from Series III C were also thicker than non-rayed leaves but not as thick as those in Series II A (pl. 28, figs. 1-3, 6, and table уп (c and m) TABLE VII SHOWING THICKNESS OF LEAVES AND LENGTH OF CELLS IN MILLIMETERS FOR RAYED AND CONTE M or 8, UERN пуно iTH TAKEN NORMA га "AC (а) Lactuca (b) Cucumis |(c) Phaseolus| (d) Ipomoea Con. Series I Con. |Series I | Con. |SeriesI| Con. | Series I G F B H G F G F G F’ H .1279| .101 |.076 | .074 |.1247| .1226 |.0938| .0814|.1565|.117 |.117 sade |.0298| .021 |.0156| .014 |.045 | .045 |.0384| .0296|.0408|.034 |.029 г epid. |.0188| .0137 |.0109 ‚0107 .0135| (—) |.0119}(—) ог|.0243|.012 | (—) , T 011 epid. |.0107| .0091 |.0107 009 .0068 .0126 |.0098| .0102|.0239|.011 |.0145 (е) (f) (g) Nicotiana | Bryophyllum| Raphanus (h) Raphanus Con. |Series I| Con. [Series I| Con. |Series I} Series 11 | Series III | Con. G H G H G F’ A B Ср | Е .143 | .1326 |.460 | .4338 |.1604| 1612 |.1582 |.158 |.149 |. 1293]. 163 ваде |.0525| .0529 | (—) | (—) 1.0495| .0635 |.0495 |.049 |.047 |.0411].038 od .0218 (—) [|.0153| (>) 1.0142] (—) |.0148 |.019 |.015 |.0134].013 нм. .011 | .0112 |.0121| .0145 |.0151| .0123 |.01176/.0103|.009 |.0117|.007 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 201 (i) Lactuca (j) Bryophyllum Series II | Series III | Control | Series II | Series III | Control а|в|с|р | Е ГА во bs № .08841.087 |.0896|.0882| .1013 1.360 |.522 |.4464|.8075| .5103 .0182|.0207|.0204!.0176! .0263 Upper ер. |.0128|.0154|.012 |.0134| .0154 |.0153|.0189|.0202|.0153| .0207 Lower epid. .0078|.0109|.0092|.0089| .0126 |.0121|.0099|.0126|.0144| .0121 * Е’, plants гауеа 8 weeks at 100 iches without a screen. t (—), lacking. (k) Cueumis (1) Coleus Series II | Series III | Control | Series П | Series ІП | Control д | в | c|n| в |А|в|сјрј Е . 1307). 13971. 1363). 127 | .125 1181. 1056]. 1232]. 1374| .138 alisade 0448). 04591. 04 453) .048 ‚03641. "0299 037 `0371 ‘0375 Оррег d 134/.0096).0117/. 0133] .0126 1.021 |.01841. `0159 . 0156 ээ Lower .007 |.0117|.007 |.0071| .0072 1.0086.0078|.01031.007 | .0112 (m) Phaseolus (n) Zea Mays Series П | Series III | Control | Series П | Series ІП | Control A B C D E A B С р Е . 1369 .1016 0915 1408}. 1402/. 1467. 1366! .1195 alisade .0518 .038 36 Upper epid. |.018 . 0111 010 .0263].0260].020 |.0296| .0274 1 012 . 0069 .0067 1.0204 . 03041. 026 . 0196] .019 (о) Nicotiana (р) Ipomoea Series II | Series Ш | Control | Series П | Series III | Control ла с |» | 2 л | в | с |» Е af 143 .1444|.1584| .145 . 1206]. 11841. 1316]. 168 | .133 Palisade 0525). 0408 .0862|.0487| .0414 |.0855|.0375|.0467|.0604| .0476 Upper ера .0218|.019 |.0148|.0162| .0207 |.0179|.0193|.0188|.0243| .0226 Lower epid. .011 |.0112|.0142|.0086| .0103 |.0168!.017 |.0176|.0212| .017 Stems.—Cross-sections from the base of non-rayed Cucumis stems seven weeks old were found to have smaller diameters than any of those from rayed stems. base of Cucumis stems in Series II A and III D were found to be a little broader, while those in Series II B and ПТС had the Similar sections from the [Vor. 15 202 ANNALS OF THE MISSOURI BOTANICAL GARDEN greatest diameter (table уш). Series II В had the thickest leaves and the greatest growth in height. All rayed Cucumis stems in Series II and III also showed larger bundles than control stems, though there was little difference between the different rayed groups. Тһе average width of the rayed bundles from the outside of the stem toward the center was 0.54 mm. and that of similar control bundles was 0.36 mm. The amount of bast tissue was about the same for control as for rayed stems. Cross-sections from the base of stems of control Nicotiana plants were also found to be smaller in diameter than most of the rayed ones. Series II A developed stems a little smaller in diameter than those of the control plants. Stems in Series II В and ПТС were larger in diameter, and those in Series III D were still larger. It will be remembered that this was the group of tobacco plants that showed the thickest leaves and the healthiest appearance (table уш). Ав to development of vascular tissue the control plants again had the thinnest vascular cylinders which were 0.36 mm. in thickness. Next in order of thickness came Series II A (0.378 mm.) followed by Series ПТС (0.505 mm.) and ПІ D (0.54 mm.). The tracheae were also smallest in the control plants and largest in Series III D with Series II A, B, and III C intermediate and equal. In all cases the walls of the tracheae were thicker in rayed plants than in non-rayed ones. When sections from the bases of rayed Zea Mays stems of the different series were measured, it was found that those in Series II A were a little smaller in diameter than those from the control stems. Series II B had stems a little larger and III D still larger than those in Series ПВ. Stems in Series ПС were the largest of all. This was also the group of plants that showed the thickest leaves and the greatest growth. There was a noticeable range of variation present in the vascular bundles of the different groups of Zea Mays plants. Rayed stems in Series ПА showed bundles smaller than those of the control stems, both as to entire bundle and as to size of vessels, but the phloem was better developed than in control stems. Series II B showed bundles of about the same size as control stems, but here the phloem was as well developed as in Series II A and the mechanical tissue much better developed than in 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 203 either the control stems or those in Series ПА. The best- developed bundles were found in Series III C. Here the phloem and mechanical tissue were very well developed. The walls of the vessels were much heavier here than in any other group of Zea Mays plants. The pith cells in the control plants and those in Series ПА were angled, while those in Series II B and III C and D were oval in shape, showing more air-spaces. The oval pith cells were also larger than corresponding angled ones. Series III C was also found to have many more layers of cells making up the cortex than any of the other groups of Zea Mays plants (pl. 33, figs. 1-8, and table уш). When sections of Coleus stem from the different groups of plants were measured, it was found that the control plants again showed smaller stems than any of the rayed plants. The stems largest in diameter were found in Series II A and B. It was these two groups also that showed the greatest growth in height (table уш). Тһе radial diameter of the vascular bundles of the control stems was 0.495 mm., while that of Series III D was 0.612 mm., that of III C, 0.62 mm., and of Series II B, 0.675 mm. This corresponds well with the fact that greatest growth in height was found in this group. Bast tissue was present about cnr. in all rayed and control stems of Coleus. Sections of Phaseolus also showed the control plants to have stems smaller in diameter than any of the rayed plants. Series II A has stems having the greatest diameter. It will be remembered that this group of Phaseolus plants also showed the thickest leaves. Series III C had stems just a little smaller in diameter than those in Series ПА (table viu). When the TABLE VIII SHOWING THICKNESS IN а ЭРЭ” ОК ВАУЕО AND NON-RAYED Series II Series III Control Plant A B С р Е cumis 5.0 5.5 5.5 5.0 4.0 Nicotiana 8.8 9.5 9.5 10.0 8.5 Zea Mays 8.0 8.8 10.7 9.5 8.5 oleus 6.0 6.0 5.5 5.5 4.8 Phaseolus 3.8 3.5 3.0 [Vor. 15 204 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN vascular cylinder in the different rayed groups was compared it was found to be by far the thickest in Series ПА and ПТС, averaging 0.54 mm. in diameter, while that of the control stems averaged 0.49 mm. PHYSIOLOGICAL Chlorophyll decomposition.—AÀ medium green 80 per cent alcoholic chlorophyll solution was made from Nicotiana leaves and put in test-tubes of pure fused silica. These were placed horizontally in white dishes and exposed to the different con- ditions, with results given in table rx. TABLE IX SHOWING THE AMOUNT OF TIME NEEDED TO DECOLORIZE CHLOROPHYLL SOLUTION 9 a.m. 12 т. Sunlight in vente АИ 12 min. min. с т.” Ж gc cues Ұзақ» a o en 4 min. шїп. Diff Bed Tight in epe revo а пи как 35 min. 18 min. At 30 inches from an unscreened lamp in diffused light... . . 39 min. 20 min. Ultra-violet p bees | with vita pnis plus diffused light| 40 min. 22 min. Sunlight outside under a screen of vita glass.............. 6 min. 3 min. Sunlight outside under a screen of ME glass....... 5 min. | 27; min. These results show plainly that ultra-violet rays do not hasten the decomposition of chlorophyll. Vita glass is thicker than quartz-lite, and hence used at close range the difference in thickness would account for the longer time required for the decomposition under vita glass in sunlight. Starch storage.—Sections of Coleus stem at the end of seven weeks showed more starch in control plants than in any of the plants rayed as in Series П and III. It was impossible, however, to distinguish between the different rayed stems. In sections of Phaseolus stem starch was present in the cortex of plants rayed as in Series II A, while similar control plants showed very little if any starch in the cortex. Determination of Py.—Lactuca plants under experiment for eight weeks as in Series I F were used for this work, the leaves and stems being determined separately. The Py of the leaves and stems of both rayed and control plants was found to be 6.0. The P, of leaves and roots of Raphanus was determined sepa- 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 205 rately, and here again the rayed and control plants responded alike, that of the leaves being 6.2 and the roots 6.0. Thus raying with ultra-violet rays seems to have no effect on the Ра of plants. Dry weights.—In the experiments carried on in 1926 to 1927 as described in Series I, which consisted of plants rayed with an unscreened lamp, dry weight determinations were made of the entire tops of Lactuca and both tops and roots of Raphanus. In all cases greater dry weight was found in the control plants. This can be well seen in the results in table x (a) and (b). TABLE X SHOWING IN GRAMS nm RY WEIGHT OF PLANTS IN SERIES I H (50 INCHES), F (100 INCHES), AND С (CONTROL) (a) Raphanus 4 weeks 4 weeks 8 weeks H | G Е | с y 1H Tops 36 | 69 .326 49 586 | 1.0 Roots 41 279 7126 7174 724 :496 (5) Tops of Lactuca plants 2 leaves 9 leaves | 2 leaves 9 leaves н | | ВО | в | G ЭС 4weeks | 018 | .244 | .805 | 2.22 | 116 | ка | 1.24 8 weeks :003 | 770 | 895 | 3.85 | 200 | 115 | 156 | 276 In the experiments carried on in 1927 to 1928 Ше dry weight was determined for fifty grams of wet weight of leaves. Rayed Zea Mays plants of Series II and III showed greater dry weight than corresponding control plants. Series II A showed the smallest dry weight of the rayed plants which was where the poorest growth in rayed plants of Series II and III was found. Lactuca plants in Series III D had the greatest dry weight. Those in Series П A and ПТС showed smaller dry weight than the control plants. Ipomoea plants in Series III D had the greatest dry weight. It was also in this group that greatest growth and thickest leaves were found. Nicotiana plants showed greatest dry weight in the control plants and the smallest in Series II A (Vou. 15 206 ANNALS OF THE MISSOURI BOTANICAL GARDEN Cucumis plants had the smallest dry weight in Series П В, and in this group were the greatest growth and thickest leaves with the largest air-spaces. Phaseolus plants in Series ПА showed the greatest dry weight and also the thickest leaves. Plants of Raphanus showed the greatest weight in Series III C and D and the next greatest in the control plants. Bryophyllum plants also had the greatest dry weight in Series Ш С and Р, though all rayed plants in Series П and П had greater weights than similar control plants. A comparison of the dry weights of the various plants will be found in table x1. TABLE XI SHORING THE DRY WEIGHT IN GRAMS PER FIFTY GBAMS OF WET WEIGHT PLANTS IN SERIES I a diy WITHOUT A SCREEN), SERIES II (SCREEN OF VITA GLASS) AND 3C M ur (SCREEN О OF QUA RTZ-LITE GLASS) Series I Series II Series III Control Plant H А В С р Е Zea Mays 6.0331 | 5.462 6.1496 | 5.8290 | 5.9232 | 5.2030 Lactuca 3.1950 | 2.2307 | 2.8396 | 2.3322 | 3.2320 | 2.7959 р 7.4775 | 7.0642 | 7.6735 | 7.7290 | 7.3554 icotiana 5.3150 | 5.5130 | 5.9240 | 5.9675 | 6.3225 Cucumis 4.8072 | 4.5494 | 4.7149 | 4.7544 | 4.8420 Phaseolus 6.9050 5.3994 6.3695 phanus 3.8845 | 3.9775 | 4.3200 | 4.3410 | 4.1361 Bryophyllum 2.8295 | 3.5100 | 3.4055 | 4.2975 | 3.7585 | 3. 5286 Ash determination.—The results were not conclusive, but they point toward an increase in ash in plants rayed with an unscreened lamp. In plants rayed with a screened lamp the ash was, in the majority of cases, less than in the control plants. In Cucumis plants showing best growth there was less ash and also smaller dry weight than in the control plants. This can be explained, however, by the presence of many large air-spaces in those leaves while in the control leaves the air-spaces were smaller. In Phaseolus the amount of ash again corresponded very well with the dry weights, there being the greatest dry weight where there was the greatest amount of ash. This also corresponded with the thickness of the leaves. For comparison of the results see table хи. 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 207 TABLE XII i SHOWING THE WEIGHT IN GRAMS OF ASH FOR 3 GRAMS OF DRY LEAF MATERIAL Plant A B C D E F Zea Mays .310 .272 .269 .295 .325 .349 Lactuca . 600 . 610 . 611 .581 .595 .565 Ipomoea .410 .302 395 392 505 Nicotiana „527 . 533 505 515 570 Cucumis ‚ 562 ‚521 505 530 599 Phaseolus .464 435 455 phanus . 638 . 630 . 643 .558 ' 789 Вгуорћу та ‚564 . 5193 .482 .430 . 540 . 610 ` The effect of ultra-violet radiation upon transpiration.—W hen leaves of Phaseolus, Cucumis, Lactuca and Coleus were placed in bottles of water, sealed with paraffin, and rayed at 50 inches from the unscreened lamp, it was found through weighings taken every 30 minutes of bottles and leaves combined that at first the rayed leaves lost as much as the controls. Then there was a time when less weight and sometimes no weight was lost by rayed leaves. After that there was a loss equalling that of the control leaves kept in darkness or in some cases surpassing it. When the stomata were examined at the end of three hours those in the rayed leaves were found to be closed, while those in the leaves kept in darkness were partly open. When the rayed and control leaves were weighed at the beginning and end of the experiment, it was found that all the rayed leaves had lost weight while the controls had remained constant (fig. 2). It will be noted that Coleus behaved a little differently than did the other leaves used. This might be explained by the fact that Coleus has stomata on the under surface only. This experiment was repeated several times with the different leaves. The petioles of leaves of Coleus Blumei var. Verschaffeltii were paraffined and the leaves placed in a horizontal position, some being rayed on the upper side, some on both sides, and others placed in darkness, Those in darkness and those rayed on the upper surface were partly wilted at the end of twelve hours while those rayed on both sides were still turgid. When weighed, however, the leaves rayed on both sides and those upon one side only were found to have lost much more weight than the leaves kept in darkness (table хит, and pl. 26, fig. 3). (Vou. 15 208 ANNALS OF THE MISSOURI BOTANICAL GARDEN ---- LOSS IN WEIGHT —-Rayed Leaf -Control Leaf 0 h 1 15 2 25 3 HOURS Fig. 2. Showing the comparison between the loss of weight of leaves rayed with an unscreened lamp and those kept in darkness. TABLE XIII SHOWING LOSS OF WEIGHT IN GRAMS OF COLEUS LEAVES WITH PARAFFINED STEMS Control Rayed on both sides | Rayed on one side Original weight 1.90 2.10 1.65 Weight after 12 hours 1.75 1.72 1.20 Loss .15 .88 .45 DISCUSSION Several points have been very clearly brought out by the foregoing experiments. All plants rayed with an unscreened 1928] ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 209 quartz mercury lamp were conspicuously injured. At a distance of 50 inches from such a lamp the injury was very great to all plants. At 100 inches away the injury was not so great for the same length of exposure, probably due to a portion of the injurious rays being absorbed by the atmosphere between the lamp and the plant. It was also evident that at this distance some plants were more resistant to ultra-violet radiation as a whole than others and that younger plants were less resistant than older ones. Тһе latter fact was particularly noticeable in Lactuca where the growth of young plants was almost completely stopped while that of older ones was only retarded. The injury was first evident in the epidermis where many of the cells if not collapsed, forming a false cuticle, were smaller than those of control leaves. After raying for a period of weeks, the in- jury to newly formed leaves was evident through the entire leaf, causing the mesophyll tissue to be more compact with fewer air-spaces and with little differentiation between the different kinds of cells. In plants such as Raphanus the contents of the palisade cells were drawn away from the upper ends of the cells, partieularly in regions where the epidermis had collapsed. These results suggest that raying with an unscreened lamp may actually retard growth in individual cells even if it does not kill them. Bailey (794), using an open arc lamp, and Kluyver (711), Ritson and Westbrook (Delf, Ritson and Westbrook, '27), using quartz mercury vapor lamps, obtained similar results though different methods of raying were used in all cases. Bailey and Kluyver found anthocyanin disappearing from rayed Coleus leaves when the color was present in the epidermis only. In the foregoing experiments when Coleus was rayed under the same general conditions, the anthocyanin pigment disappeared also from the palisade cells of the leaves and Їгога the entire stem tips. Until recently the penetration of the short ultra-violet rays was thought to be very slight. In fact a layer of skin was said to inhibit their passage. Macht, Anderson, and Bell (728), however, using living anesthetized animals have shown that rays as short as 313 ши penetrate into the peritoneal cavity of a rabbit. [Vor. 15 210 ANNALS OF THE MISSOURI BOTANICAL GARDEN Therefore it might not be unusual for rays to penetrate through plant epidermis into the palisade cells or into the cortex of a stem tip, either destroying the pigment or preventing its formation. Green (’97) has suggested that chlorophyll might act as a screen absorbing injurious rays. This supposition is strengthened by the results of experiments on chlorophyll in this paper where ultra-violet rays were found to have very little, if any, effect upon chlorophyll decomposition. If there is a screening action of chlorophyll, it might not have been evident here due to the small number of chloroplastids present in the cortex of the stem. On the other hand, the rays might not have penetrated deep enough to cause direct action, but may have set up chemical reaction which induced the decomposition of anthocyanin or prevented its formation. Sheard and Higgins (’27) found that in general rays of 270 to 320 up delayed the time of germination and lessened the rate of growth, but that rays of 320-390 yy were effective in promoting growth. In the experiments in this paper where the lamp was screened by vita glass which cut out rays below 290 uu, there were no lesions though the newly formed leaves of Lactuca, Raphanus, and Coleus were thinner. In the other plants used the leaves were either of the same or greater thickness than control leaves. When the lamp was screened by quartz-lite glass which cut out rays shorter than 310 uu there was also no evidence of lesions though the same three plants again had thinner leaves, while all others had much thicker leaves than the control plants. In nearly all cases where leaves were found to be thicker when rayed by a screened lamp, the plants themselves were found to be taller with larger stems and more numerous leaves. In no cases was flower production retarded, and in Cucumis and Phaseolus it was slightly increased. Sometimes the increase in size took the form of more numerous and larger air-spaces with only slightly larger cells. This was true of both the stems and leaves of Cucumis and Nicotiana and of the stems of Zea Mays. In other cases there were more air- spaces but when this occurred the palisade cells were very much larger. This was very noticeable in Ipomoea and Phaseolus. In 1928 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 211 the leaves of Zea Mays the increased growth was evident only in the form of larger cells. Coleus plants rayed with a lamp screened by vita glass or quartz- lite glass showed a marked increase in growth over control plants, though the thickness of the leaves was inversely proportional to their increase in height and number. This might indicate in- cipient injury to the leaves with a possible stimulatory effect upon the plant as a whole. In addition there was no loss of red color here as there had been when an unscreened lamp was used. The results with the use of screens correspond well with those of McCrea (’27), where increased growth was obtained in Digitalis by the use of vita glass instead of ordinary glass in a greenhouse, and with those of Tsuji (718), who produced larger and juicier pineapples by raying them in the field with a weak ultra-violet lamp. When Phaseolus plants with both cotyledons removed were rayed either by a screened or an unscreened lamp there was a re- tardation in growth compared with corresponding non-rayed plants. When only one cotyledon was removed from Phaseolus plants, raying with a screened lamp produced a slight increase in growth over corresponding non-rayed plants. This compares well with the work of Westbrook (Delf, Ritson, and Westbrook, ’27) where different lengths of day were used in addition to a short daily raying with an unscreened lamp. Іп all cases the injury was greater the shorter the day. These results and the fact that older Lactuca plants were more resistant than young ones may indicate the possibility that the presence of sufficient food may at least partially overcome the injurious effects of raying. Clement (’26) found that apples that had been rayed stayed turgid longer than non-rayed ones. Likewise Tsuji (718) ob- served that stalks of banana plants that were rayed after being cut kept fresh many days longer than non-rayed ones. The writer has found that if the petioles of Coleus leaves were dipped in paraffin and the leaves then rayed with an unscreened lamp first on one side and then on the other for a period of one and a half hours each, they remained turgid at the end of twelve hours, while the control leaves were badly wilted. If, however, the leaves were rayed only on one side the leaves were much more [Vor. 15 212 ANNALS OF THE MISSOURI BOTANICAL GARDEN wilted than the control leaves. All these observations would tend to indicate that a false cuticle is formed wherever plant tissue is rayed heavily with ultra-violet, thus greatly retarding the rate of water loss from the internal tissues. In the foregoing experiments, using a wide variety of plants in sufficient numbers to avoid individual differences, and using two well-known ultra-violet glasses as screens, the writer has reached the conclusion that increased growth can be obtained in many groups of plants by the daily use of a quartz mercury vapor lamp screened to cut out the harmful short rays. However, results point to the supposition that each variety of plant has its own ultra-violet requirement for best growth and that this can be determined only by experiment. SUMMARY 1. Raying with an unscreened quartz mercury vapor lamp caused injury in all plants used. 2. Raying with a lamp screened by vita glass was beneficial for some plants, while it produced little visible effect in others. When examined anatomically no lesions were present, but in some cases there was a slight retardation of growth. 3. Raying with a lamp screened by quartz-lite glass injured none and benefited many of the plants. In some cases, however, the benefit was less than when vita glass was used. 4. Except for Raphanus and possibly Lactuca the healthiest- appearing plants were among those rayed with a screened lamp, although the distance from the light and the screen promoting best growth differed for different plants. 5. Raying with a screened lamp increased flower production slightly. 6. Plants rayed for a period of weeks with an unscreened lamp developed leaves which were thinner than those of corresponding non-rayed plants, the decrease in thickness being due to a partial or complete collapse of the upper epidermal cells, a lack of differ- entiation of the palisade layer, and a decrease in the number and size of the air-spaces present in the mesophyll tissue. 7. With the exception of Coleus, Raphanus, and Lactuca, leaves of plants rayed with a screened lamp were in general thicker than 1928] - ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION 213 corresponding leaves from control plants, though the particular screen and distance from the lamp promoting the formation of the thickest leaves differed for different plants. The increase in thickness was due either to increase in size of cells or to increase in number and size of air-spaces or to both. 8. Тһе stems гауеа with a screened lamp were greater in diam- eter and contained better-developed vascular bundles than non- rayed ones. 9. A limitation of the amount of available food emphasizes the injurious effect of ultra-violet rays. 10. Ultra-violet radiation had very little, if any, effect upon the decomposition of chlorophyll and thus very little effect upon the photosynthetic apparatus. 11. Ultra-violet radiation had no effect upon the Py of the plants used. These results again emphasize the fact that each plant has its own ultra-violet requirement for best growth which can be de- termined only by experiment. ACKNOWLEDGMENTS The writer wishes to express sincere appreciation to Dr. B. M. Duggar, Professor of Physiological and Applied Botany, University of Wisconsin, formerly Professor of Plant Physiology in the Henry Shaw School of Botany of Washington University, for suggesting this problem and under whose guidance the early part of this work was carried out; to Dr. Е. 5. Reynolds, Physiologist to the Missouri Botanical Garden, for his advice and helpful criticisms concerning the work; to Dr. C. F. Hagenow, of the Physics Depart- ment of Washington University, for spectral photographs; to Dr. LeRoy McMaster, of the Chemistry Department of Washington University, for the use of laboratory and apparatus; and to Dr. G. T. Moore, Director of the Missouri Botanical Garden, for the privileges and facilities of that institution. BIBLIOGRAPHY Bailey, L. Н. (94). Electricity and plant growing. Mass. Hort. Soc. Trans. 1894: 1-28. 1894. Barr, С. E. and У. Т. Bovie (23). Ultra-violet cytolysis of protoplasm. Jour. Morph. 38: 295-300. 1923. [Vor. 15 214 ANNALS OF THE MISSOURI BOTANICAL GARDEN Bazzoni, С. B. (14). The destruction г гэ through the action of light. . Jour. Pub. Health 4: 915-992. Beeskow, H. C. (’27). Some ован в, цав of ultra-violet sade on plants. Report given at Nashville meeting of Am. Soc. Plant Physiol. Dec. 1927. Воме, W.T., and G. A. Daland (23). New experiments on the актива of proto- ` plasm to heat by exposure to light of short wave-length. Am. Jour. Physiol. 66: 55-66. 1923. жону Matilda (26). Effect of light of different wave lengths on penetration of — dibromo phenol indophenol into Valonia. Soc. Exp. Biol. & Med. Proc : 576-577. 1926. Bure, W. E. (17). Action of ultra-violet radiation in killing living cells such as ia. Am. Jour. Physiol. 43: 429-432. 27: Cala J. 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The influence of selective and general radia- ns a a quartz mercury lamp upon germination and growth of seeds. Science 5: 282-284. 1927. оше, А. (11). Über den Einfluss der ultra-violetten Strahlen auf die Vegeta- tion. Centralbl. f. Bakt. П Abt. 31: 477. 1911. , (15). Über die Bedeutung der Einwirkung der ultravioletten Strahlen auf die photochemische Synthese der Kohlenhydrate in der Chlorophyll haltigen Zelle. Zentralbl. f. Biochem. und Biophys. 18: 370. 5. Tanner, F. W., and E. Ryder (28). Action of ultra-violet light on yeast-like fungi. Bot. Gaz. 75: 309-317. 1923. Tshuhotine, S. (23). Sur la mecanisme de l'action des rayons ultra violets sur la Inst. Pasteur, Ann. 35: 321-325. 1923. Tsuji (18). Stimulation in growth of sugar cane — increase of percentage of sugar on exposure to ultra-violet rays. La. Planter 60: 413. 1918. Ursprung, A., und С. Blum (17). Uber die Schadigkeit ultravioletten Strahlen. Ber. d. деш. Bot. Сев. 35: 385-402. 1917. (Мог. 15, 1928) 216 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 21 (SERIES I F, H AND G) Fig. 1. Cucumis sativus. A. Plant rayed four weeks at 100 inches from the light without a screen. B. Plant not rayed. Fig. 2. Cucumis sativus. A. Plant not raye B. Plant rayed for eight weeks as in fig. 1 A. B. Plants rayed for eight weeks at 100 inches from the light without а screen. C. Plants rayed for eight weeks at 50 inches from the light without a screen. Fig. 4. Raphanus sativus. A. Plant rayed for eight weeks at 100 inches as in fig. 3 B. Fig. 5. Raphanus sativus. А. Plant и. - four weeks at 100 inches as in fig. 3 B. B. Plant not r Fig. 6. Ipomoea g^ atat А. Plant rayed for vd weeks at 100 inches from the light without а screen. B. Plant not rayed. Fig. 7. Raphanus sativus. А. Plant not rayed. B. Plant rayed for eight weeks as in fig. 3 B. ANN. Мо. Bor. Garb., Vor. 15, 1928 PLATE 21 ELTINGE—EFFECT СЕ ULTRA-VIOLET RADIATION [Vor. 15, 1928] 218 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 22 (SERIES I H, F AND G) Fig. 1. Lactuca sativa (nine leaves) rayed for four weeks. A. Plant rayed at 50 inches from the light without a screen. B. Plant rayed at 100 inches from the light without a screen. C. Plant not rayed. Fig. 2. Lactuca sativa (two leaves) rayed for eight weeks. A. Plant rayed at 50 inches from the light without a screen. B. Plant rayed at 100 inches from the light without a screen. C. Plant not rayed. Fig. 3. Lactuca sativa (two leaves) rayed for four weeks. A. Plant rayed at 50 inches from the light. B. Plant rayed at 100 inches from the light without a screen. C. Plant not rayed. Fig. 4. Coleus Blumei var. “Spotted Gem." А. Plant rayed for two weeks at 50 inches from the light without а screen. B. Plant not rayed. Fig. 5. Coleus Blumei vars. Verschaffeltit and “Spotted Gem." А. Var “Spotted Gem" rayed for eight weeks at 100 inches from the light with- out a screen and then allowed to recover in the greenhouse for four weeks. B. Var. “Spotted Gem" not rayed. C. Var. Verschaffeltit treated the same as in fig. 5 A. D. Var. Verschaffeltii not rayed. ANN. Мо. Вот. Garb., Vor. 15, 1928 PLATE 22 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION [У ог, 15, 1928] 220 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 23 (SERIES I F AND G) Fig. 1. Coleus Blumei vars. “Spotted Gem" and Verschaffeltii. А. юун В. ри Сет” rayed for four weeks at 100 inches from the light with- out a B. Var, "Spotted Gem” not rayed. C. Var Verschaffeltii rayed under the same conditions as Spotted Gem." D. Var. Verschaffeltii not rayed. Fig. 2. Coleus Blumei var. “Spotted Gem” and Verschaffeltii. А. Var. Verschaffeltii not rayed. B. Var. Verschaffeltii rayed for seven weeks at 100 inches from the light with- out а screen C. Var. “Spotted Gem” rayed for seven weeks under the same conditions as in fig. 2 B. D. Var. “Spotted Gem" not rayed. Fig. 3. Coleus Blumei var. ‘Defiance ” and “Trailing Queen." A. г. “Defiance” not rayed. B. Var. " Defiance" rayed for four weeks under the same conditions as the plants in fig. 1 A. C. Var. “Trailing Queen” not rayed. D. Var. “Trailing Queen" rayed for four weeks under the same conditions as fig. 1 A. Fig. 4. nre Blumei var. “Defiance” and “Trailing Queen." A. г. “Defiance” not raye B. Va ar. “ Defiance" rayed hr seven weeks under the same conditions as fig. 1 A. C. Var. “Trailing Queen" rayed for seven weeks under the same conditions as fig. 1 A. D. Var. “Trailing Queen" not rayed. ANN. Mo. Вот. Санр., Vor. 15, 1928 PLATH 25 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION ГУ от. 15, 1928 222 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 24 (SERIES II AND III) Fig. 1. Cucumis sativus. A. Plant not rayed. B. Plant rayed for seven weeks at 100 inches, using a screen of quartz-lite glass. C. Plant rayed for seven weeks at 50 inches, using a screen of quartz-lite glass. D. Plant rayed for seven weeks at 100 inches, using a screen of vita glass. B. Plant rayed for seven weeks at 100 inches, using a screen of quartz-lite glass. C. Plant rayed for seven weeks at 50 inches, using a screen of quartz-lite glass. D. Plant rayed for seven weeks at 100 inches, using a screen of vita glass. E. Plant rayed for seven weeks at 50 inches, using a screen of vita glass. B. Plant rayed for seven weeks at 100 inches, using a screen of quartz-lite glass. C. Plant rayed for seven weeks at 50 inches, using a screen of quartz-lite glass. D. Plant rayed for seven weeks at 100 inches, using a screen of vita glass. E. Plant rayed for seven weeks at 50 inches, using a screen of vita glass. Fig. 4. Lactuca sativa. A. Plant not rayed. B. Plant rayed for seven weeks at 100 inches, using a screen of quartz-lite glass. C. Plant rayed for seven weeks at 50 inches, using a screen of quartz-lite glass. D. Plant rayed for seven weeks at 100 inches, using a screen of vita glass E. Plant rayed for seven weeks at 50 inches, using a screen of vita glass. ANN. Mo. Bor. Garp., Vor. 15, 1928 PLATE 24 = ELTINGE—EFFECT ОЕ ULTRA-VIOLET RADIATION 224 Е (Мог. 15, 1928) ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 25 (SERIES П AND ІП) g. 1. Raphanus sativus. A. Plant not rayed B. Plant rayed for seven weeks at 100 inches from the light, using a screen of quartz-lite glass C. Plant ip for seven weeks at 50 inches from the light, using a screen of quartz-lite gl D. Plant rayed for seven weeks at 100 inches from the light, using a screen of vita glass. Е. Plant rayed for seven weeks at 50 inches from the light, using a screen of vita glass. Fig. 2. Coleus Blumei var. “Spotted Gem." A. Plant rayed for seven weeks at 50 inches from the light, using a screen of vita glass. B. Plant rayed for seven weeks at 100 inches from the light, using a screen of vita glass. C. Plant rayed for seven weeks at 50 inches from the light, using a screen of quartz-lite glass D. Plant фена Гог seven weeks at 100 inches from the light, using a screen of quartz-lite glass E. Plant not ra Fig. 3. Coleus Blumei var. Verschaffeltit. E Fig. A A. lm rayed for seven weeks at 50 inches from the light, using a screen of vita glas B. Plant rayed for seven weeks at 100 inches from the light, using а screen of vita glass. | С. Plant rayed for seven weeks at 50 inches from the light, using а screen of quartz-lite glass. D. Plant е на Гог seven weeks at 100 inches from the light, using a screen of quartz-lite glass Plant not ra g. 4. Бонн pinnatum. lant rayed for seven weeks at 50 inches from the light, using a screen of vita glass. B. Plant rayed for seven weeks at 100 inches from the light, using a screen of vita glass. C. Plant rayed for seven weeks at 50 inches from the light, using a screen of quartz-lite glass D. Plant rayed for seven weeks at 100 inches from the light, using a screen of quartz-lite glass E. Plant not rayed. ~ Ann. Mo. Вот. Garp., Vor. 15, 1928 ELTINGE EFFECT OF ULTRA-VIOLET RADIATION 226 ANNALS OF THE MISSOURI BOTANICAL GARDE [ Уот, 15, 1928] N EXPLANATION OF PLATE PLATE 26 ең 1. Гасїшса sativa (fifteen leave . Plant rayed for eight weeks at 100 inches from ап unscreened lamp. ^ Plant not rayed. Fig. 2. Coleus Blumei var. Verschaffeltit. A. Plant rayed for thirty seconds each day at 100 inches from an unscreened lamp B. Plant not rayed. Fig. 3. Leaves of Coleus Blumet var. Verschaffeltii, with petioles paraffined. А. Leaf twelve hours after it had been rayed for 1!4 hours on each surface at thirty inches from an unscreened lam B. Leaf twelve hours after it had been saved E 11% hours upon the upper surface at thirty inches from an unscreened lam C. Unrayed нь after twelve hours. Fig. 4. Zea Мау A. Plant not ra B. Plant rayed pe six weeks at 100 inches from an unscreened lamp. Fig. 5. Bryophyllum pinnatum. А. Plant not гауе B. Plant rayed for six weeks at, 100 inches from an unscreened lamp. Fig. 6. Phaseolus aris. A. Plant rayed bot four weeks at 100 inches from an unscreened lamp. B. Plant not rayed. ANN. Mo. Вот. Garb., Vor. 15, 1928 PLATE 26 Ф еч” Јо :w ж T ЕЗ Две г. = ) Cit ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION ГУ он, 15, 1928] 228 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 27 Camera-lucida drawings of equal magnification, using 4-mm. objective and 10 Х eyepiece. Fig. 1. Leaf of Lactuca sativa not rayed. Fig. 2. Leaf of Lactuca sativa rayed for four weeks at 100 inches from the light without a screen. Fig Leaf of Lactuca sativa rayed for eight weeks at 100 inches from the light without a screen Fig. 4. Leaf of Lactuca sativa rayed for four weeks at 50 inches from the light without a screen Fig. 5. Leaf of Nicotiana Тађасит rayed for seven weeks at 100 inches from the light, using a screen of quartz-lite glass. Fig. 6. Leaf of Nicotiana Tabacum not rayed. Fig. 7. Leaf of Nicotiana Tabacum rayed for four weeks at 50 inches from the light without a screen. Ann. Mo. Вот. Garb., Vor. 15, 1928 PLATE 27 ELTINGE-—EFFECT OF ULTRA-VIOLET RADIATION А а Мања у: КАКА ОМА см АМИ ье РА сој 726 TO х, ^ An RES NER ель Ра д РАҚ Lu rx [Vor. 15, 1928] 230 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 28 ее drawings of equal magnification, using 4-mm. objective and 10 X ey Fig. ES Leaf of Phaseolus vulgaris not rayed. Fig. 2. Leaf of Phaseolus vulgaris rayed for four weeks at 100 inches from the light without a screen. Fig. 3. Leaf of Phaseolus vulgaris = for seven weeks at 50 inches from the light, using a screen of quartz-lite g Fig. 4. Leaf of Cucumis sativus not г raved. Fig. 5. Leaf of Cucumis sativus rayed for four weeks at 100 inches from the light without a screen. Fig. 6. Leaf of Phaseolus — rayed for seven weeks at 50 inches from the light, using a screen of vita g ANN. Mo. Bor. Слво., Vor. 15, 1928 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION PLATE 28 [У ог. 15, 1928] 232 ANNALS OF THE MISSOURI BOTANICAL GARDE EXPLANATION OF PLATE PLATE 29 Camera-lucida drawings of equal magnification, using 4-mm. objective and 10 X eyepiece Fig. 1. Leaf of Cucumis sativus unra Fig. 2. Leaf of Cucumis sativus юэ fot seven weeks at 100 inches from the light, using a screen of vita glass. Fig. 3. Leaf of Cucumis sativus rayed for seven weeks at 100 inches from the light, using a screen of quartz-lite glass Fig. 4. Leaf of Ipomoea Batatas unrayed. Fig. 5. Leaf of Ipomoea Batatas rayed for seven weeks at 100 inches from the light, using a screen of quartz-lite E Fig. 6. Leaf of Ipomoea В at hes f the light without а screen. ANN. Mo. Bor. Савр., Vor. 15, 1928 PLATE 29 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION (Мог. 15, 1928] 234 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 30 Camera-lucida drawings of equal magnification, using 4-mm. objective and 10 X eyepiece Fig. 1. Lest of Coleus Blumei var. Verschaffeltii rayed for seven weeks at 50 inches from the light, using a screen of vita Leaf of Coleus Blumei var. Verschafelti rayed for seven weeks at 100 inches from the light, using а screen of vita glass. Fig. 3. Leaf of Coleus Blumei var. Verschaffei not rayed. Fig. 4. Leaf of Coleus Blumei var. Verschaffeltii rayed for seven weeks at 50 — from the light, using а screen of quartz-lite glass . 5. Leaf of Coleus Blumei var. Verschaffeltii шэг for seven weeks at 100 бый from the light, using a screen of quartz-lite glass. Ann. Mo. Вот. Garb., Vor. 15, 1928 PLATE 20 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION [Vor. 15, 1928] 236 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 31 Fig. 1. Diagram of the cross-section ийн а A of Coleus Blumei, the stippling "20 the normal distribution of red с Diagram of the cross-section Нэн а Uo of Coleus Blumei, showing the distribution ын red color at Ше end of the fifth raying at 50 inches from the light without а 8 Fig. 3. P am of the cross-section of а stem of Coleus Blumei, showing the distribution of E eolor at the end of the tenth raying at 50 inches from the light without a scree Fig. 4. Dabo. of a leaf of Coleus Blumei, showing normal distribution of red color. ig. 5. Cross-section of a leaf of Coleus Blumei, showing the distribution of red color after ten rayings at 50 inches from the light without a screen. Drawn to the same scale as fig. 4. ANN. Mo. Вот. Garb., Vor. 15, 1928 PLATE 31 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION ГУ от, 15, 1928] 238 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 32 Camera-lucida drawings of equal magnification, using a 4-mm. objective and 10 X eyepiece. Fig. 1. Leaf of Zea Mays unrayed. Fig. 2. Leaf of Zea Mays rayed for seven weeks at 50 inches from the light, lass. using a screen of vita g Fi Leaf of Zea Mays rayed for seven weeks at 50 inches from the light, using a screen of quartz-lite glass Fig. 4. Leaf of Zea Mays faved for seven weeks at 100 inches from the light, using а screen of quartz-lite glass Fig of Zea Mays кува at 100 inches from the light for seven weeks, using a screen of vita g Fi Leaf of Zea Mays rayed for seven weeks at 100 inches from the light, using no screen. PLATE 32 ANN. Mo. Bor. Слво., Vor. 15, 1928 ELTINGE—EFFECT OF ULTRA-VIOLET RADIATION [Vor. 15, 1928] 240 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 33 Camera-lucida drawings of equal magnification, using 16-mm. objective and 10 х eyepiece. Corresponding bundles were used in all cases (sixth bundle from the epidermis). Fig. 1. Cross-section of a fibrovascular bundle of Zea Mays from an unrayed stem. Fig. 2. Cross-section of a fibrovascular bundle of Zea Mays from a stem rayed for seven weeks at 50 inches from the light, using a screen of quartz-lite glass. Fig. 3. Cross-section of a fibrovascular bundle of Zea Mays from a stem rayed for seven weeks at 50 inches from the light, using a screen of vita glass Fig. 4. Cross-section of a fibrovascular bundle of Zea Mays from a stem rayed for seven weeks at 100 inches from the light, using a screen of vita glass. Fig. 5. The amount of cortex present in a stem of Zea Mays rayed for seven weeks at 50 inches from the light, using a screen of quartz-lite glass Fi The amount of cortex present in a stem of Zea Mays кте Гог веуеп weeks at 50 inches from the light, using a screen of vita glass The amount of cortex present in a stem of ea Mays rayed for seven weeks at 100 inches from the light, using a screen of vita glass Fig. 8. The amount of cortex present in an unrayed stem of Zea Mays. PLATE 33 15, 1928 ANN. Мо. Вот. Garb., Vor. = + 55 ДА ва: Pu Ф Фа DES NA 0: C ы = в SE? \7 Ф, [7 252 RUS TR = pe, 6 R @ A 225 чер 598 Нух TION RADIA ELTINGE—EFFECT OF ULTRA-VIOLET 22... Ду ” а T гай ОЛ. ГЭГХ "ЭГ ха а SY VR. f Mos РАСА у Missouri Botanical Garden Vol. 15 SEPTEMBER, 1928 No. 3 THE PROBLEM OF SPECIES IN THE NORTHERN BLUE FLAGS, IRIS VERSICOLOR L. AND IRIS VIRGINICA L. EDGAR ANDERSON Geneticist to the Missouri Botanical Garden Assistant Professor of Botany in the Henry Shaw School of Botany of Washington University TABLE OF CONTENTS С st is ccs tees С г. рН 241 І, тұзу and morphology... . . 2... Уан ноаен 247 ПІ. Intra-specific variation in i ris versicolor and Iris тітдітіса............... 258 IV. Hybridization of Iris versicolor and Iris йїтййтїса....................... 299 Т..Бариаагул оозе x КЕ а о сао аа ое аа 304 VE Гинэй, уг хээл ээ зэ хээ ЭМ ИН ЫЗА ко ог «асна 806 ҮЙ. БИО ХАОЦҮ, үх 24 аре rrr окр Ras лера, 812 I. INTRODUCTION The problem of species is a dual one. It asks two questions: What are species? How have they originated? In studying the nature of species we are first of all concerned with the extent to which the species of the orthodox taxonomists reflect actual divisions among plants and animals. Do such species, in the main, rest upon discrete natural groups of individuals or do they represent merely an artificial cataloguing device, ordering up, as best they may, the myriad forms occurring in nature. It might be supposed that upon so fundamental a problem biologists would have come to some working agreement. Even the most casual survey of recent literature will show that this is far from being the case. While several of those who have committed themselves to print on the subject may use such phrases as ‘“‘modern biologists recognize this to be the сазе,” “all geneticists agree, etc., etc.," it is a noteworthy fact that no two of them аге of the same shade of opinion. At one extreme are those who believe with Lotsy (716) that, ‘‘The species is merely а Ann. Мо. Вот Garp., Vor. 15, 1928 (241) 4Ё [Vor. 15 242 ANNALS OF THE MISSOURI BOTANICAL GARDEN conception of the human mind and a very primitive anthropocen- tric conception in the bargain. * * * The species of the taxonomist is the comparative insignificant rest of large swarms of individuals which arose from the cross of two parents. * * * Species in the present taxonomic sense do not exist." On the other hand, we have such statements as Harper’s (’23) ‘‘ Linnean species do, by and large, constitute recognizable groups of more or less freely inter-breeding individuals. Only extremists deny the possibility of segregating and recognizing such units." The disagreement as to species is just as extreme in regard to their size as in regard to their nature. If groups corresponding to our species do occur in nature, do they fit most readily into the species of the orthodox taxonomists (what we may call Linnean species for want of a better name) or would some other unit better express the relationships which we actually find among indivi- duals? What are we to do with the vexed question of true-breed- ing sub-groups (variously termed micro-species, Jordanons, iso- reagents) whose existence has been demonstrated for numerous Linnean species? Are we to retain them as significant sub-groups within the Linnean species, are we to treat them as of even greater importance, or are we to cast them out altogether? It should ђе а relatively simple matter to answer these questions for any one species. A comprehensive survey of variation within the species over its entire range would show whether it were an independent unit or whether it merged into other forms; whether there were recognizable sub-groups within it; and what inter- relationships obtained between the individuals which go to make up these sub-groups and the species itself. However, such an intensive study should be able to contribute to other questions besides the nature of species. It should offer valuable evidence for the more popular question of their origin. During the last quarter of a century an attempt has been made to study evolution experimentally. In the course of these ex- periments two types of mutations have been observed in our laboratories and breeding plots. They have been thought to be the same sort of changes which, operating in the past, have brought about the evolution of our present-day forms. The first type of mutation has been shown to be due to changes which take 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 243 place at a particular point in the germ-plasm. Since only a single gene is affected, such changes have been called gene- mutations. The second type of mutation has been shown to be due to re-alignments of the germinal material, to duplications of chro- mosomes or whole sets of chromosomes. The morphological results of these two types of mutation are quite dissimilar. An intensive and extensive survey of variation within a single species should therefore be able to demonstrate which of these two processes has been most active in causing progressive changes within that species. Such a study should, in other words, enable us to evaluate the evolutionary importance of gene-mutations and chromosomal re-alignments. The present study is just such an intensive and extensive survey of two closely related species. It is an attempt to present a fairly complete picture of the variation within two natural groups of individuals over their entire range. It has as yet been almost purely morphological in scope. Though the morphologi- cal differences between individuals and groups of individuals undoubtedly rest upon basic physico-chemical ones, our knowledge of these physiological differences is as yet too incomplete, among the flowering plants, to possess much phylogenetical significance. So far as is known, no such complete survey of variation within a species or group of species has ever before been made with plant material. It is an ambitious attempt for a single individual unless the problem be made as simple as possible. If we are to learn anything about the ultimate nature of species we must first of all reduce the problem to the simplest possible terms and study | a few easily recognized, well-differentiated species. The group to be studied should therefore possess few sub-groups and inter- grading forms. Unfortunately those species which have been selected for intensive study in the past have been chosen by reason of their very complexity. The northern blue flags (Iris versicolor of the seventh edition of Gray’s ‘Manual’) were accordingly chosen for the study, prima- rily, because they were a comparatively simple, stable, and well- marked group. They possessed certain other features which have materially reduced the labor involved in locating and study- (Мог, 15 244 ANNALS OF THE MISSOURI BOTANICAL GARDEN ing a large number of individuals. They are common, conspicu- ous, colonial, and perennial. Though the merits of these charac- ters are practically self-evident, it may not be out of place to call attention to the colonial nature of 1748 versicolor L. and its close relatives. Plants of these species are seldom found as isolated individuals but usually occur in colonies of a few to several hun- dred plants. Nor are these colonies merely the vegetative off- spring of asingle plant as is the case with so many colony-forming species. The individuals vary so strikingly in the size, shape, and color of their flowers, that at blooming time the limits of a single clone can readily be determined. These seldom exceed a few square feet, though exceptionally large clones do sometimes occur, as will be described below. It was the truly colonial nature of the blue flags which was particularly useful in the pres- ent study, making possible the examination of large numbers of plants in each locality with a minimum of effort. An attempt was made to visit as much as possible of the range of the northern blue flags during their flowering season. Numer- ous colonies were studied in detail and measurements made on twenty to fifty individuals of the characters which had been selected for study. Representative plants were sent back from each of the colonies and established in an experimental plot at the Missouri Botanical Garden, where they were made the subject of genetical, morphological, and cytological studies. In 1923 collections were made in central Michigan. In the spring of 1924 a trip was taken through northern Missouri, eastern Iowa, southern Wisconsin, central Michigan, western New York, and central New Hampshire. In 1925 collections were made in central Missouri, northwestern Arkansas, northern Ohio, including the Bass Islands, western and central New York, and southern Vermont. Fruiting material was collected that fall from central and northern New Hampshire and eastern Michigan. In the spring of 1926 a trip was taken through southern Illinois, southern Missouri, eastern Arkansas, western Tennessee, and central Kentucky. In June, collections were made in Missouri, Michigan, northern Vermont, Ottawa, Canada, and at Lake Timagami, in northern Ontario, Canada. In the early spring of 1927 another southern trip was taken, principally 1928 ANDERSON—PROBLEM OF SPECIES IN IRIS 245 to collect material from Mississippi and Alabama. In June of that year collections were made in Ohio, Maryland, Pennsylvania, New York, Ontario, northern Michigan, and Wisconsin. Іп 1928 95 90 : 85 80 75 — р -a А Р „5 ws 4% ^ + "12 , ғ ҒА | | КА! РА diu р 45 pA Ua е f^ dti : А „<< ~ № у; A”: 4.. МЕ ; V 22 >. 4 y PS i 2127 А T » Р, s 9^ | ~ Жм. --..Q ac VW e 9o г | < j | ЮУ 7 Е № 5 c ч - 39 LAT n ^) { о - ~ = i СА "Уу 4 | | 1 versicolor esl о а: g ! Y Ё | Оз INICS · M А / n оч » hb - Р . 2 Фу | ЖАҒАСЫЗ. 95 90 55 P Fig. 1. Localities at which 1. versicolor and 1. virginica have been studied. North and South Carolina and eastern Tennessee were visited in early May. With the exception of the extreme north the terri- tory has now been quite thoroughly covered as shown by the map in fig. 1. Мог. 15 246 ANNALS OF THE MISSOURI BOTANICAL GARDEN Extensive collections of flowering material preserved in alcohol and of ripe seed capsules were made at several points by Mr. R. E. Woodson, Jr. and by the writer. The work on the fresh and preserved material has been supplemented by taxonomic studies in the herbarium of the Missouri Botanical Garden and at various other herbaria. ACKNOWLEDGEMENTS It is scarcely possible to carry on a study involving geographical distribution without enlisting the help of a very large number of people. For aid in assembling information and material for study it is a pleasure to acknowledge the cooperation of those whose names are listed below, and of several others who have been inadvertently overlooked: Dr. J. R. McLeland, R. W. Shreve, A. N. Rood, R. 8. Sturte- vant, W. M. Boswell, Dr. A. C. Kinsey, Dr. P. C. Mangelsdorf, Prof. J. С. Jack, Dr. J. М. Couch, E. A. Mosley, Mr. Holtiwanger, Dr. O. E. Jennings, John MacDougal, A. F. Satterthwaite, M. E. Mains, Miss G. V. Butcher, Mrs. Jasper Blackburn, A. C. Anderson, Herbert Stoddard. Miss Katherine Perkins, Mrs. Will Spear, Miss Edith Mason, J. B. Stevenson, E. J. Layton, Henry Edmonds, J. E. Gilmour, J. Steyermarck, John H. Rush, Miss Martha Beardsley, Mrs. Dorothy M. Anderson, Phil Rau, Miss Mildred Mathias, Miss Katherine H. Litch. Sincere appreciation is due the curators of the United States National Herbarium, the Gray Herbarium of Harvard University, the Dillenian Herbarium, the herbaria of the Field Museum, of the University of Wisconsin, of the Michigan State College, of the Philadelphia Academy of Natural Sciences, of the Linnean Society, and the British Museum, for the opportunity of studying material from these institutions. Thanks are also due Professor J. Paul Goode, of the University of Chicago, for permission to use his Homolosine Equal Area Projection Map (fig. 1.) For much advice and helpful information in connection with the taxonomic aspects of the problem, the author is deeply indebted to his colleague, Dr. J. М. Greenman. For watchful care of the experimental garden during his absence he is pleased to thank Mr. P. A. Kohl. The study would have been less complete in many ways but for the assistance of Mr. R. E. Woodson, Jr. in 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 247 making the collections and tabulating the data. It is a pleasure to acknowledge his helpful companionship and material aid. II. Taxonomy AND MORPHOLOGY HISTORY OF THE SPECIES It has been found that what commonly passes for Iris versi- color L. is made up of two species. One, inhabiting New England, New York, Pennsylvania, northern Ontario, and northern Mich- igan, has lanceolate petals much shorter than the sepals, a short ovary, and shiny D-shaped seeds. The other extends in the middle-western states from the Great Lakes to the Gulf of Mexico and up the Atlantic seaboard to southern Virginia. It has obovate-spatulate petals which are nearly as long as the sepals, a long ovary, and dull, round or D-shaped seeds. As will be brought out in Parts III and IV of this paper, the species are wholly distinct and crosses between them are partially sterile. Their distribution is shown in more detail in fig. 1. It now becomes necessary to determine their relation to the species named by Linnaeus. In the first edition of the ‘Species Plantarum’ he named, as numbers 10 and 11, two similar Amer- ican irises. To the first, which he named Iris versicolor, ће referred numbers 187 and 188 of Dillenius and the Iris latifolia Virginiana etc. of Ehret. Тһе second he named 1718 virginica, basing it upon Gronovius’ Iris corollis imberbibus, etc. The two species of Iris studied by the writer are very similar to each other and, like many other species of the genus, are difficult of recogni- tion in ordinary herbarium specimens. Were it not for the fact that we have such ample material it might be difficult, if not impossible, to determine the relation between the names given by Linnaeus and these two common species of eastern North America. Fortunately there are such good illustrations and such excellent herbarium specimens that there can be no doubt that the common blue flag of the East and North is identical with [тїз versicolor of Linnaeus and that the species of the South and Middle West is identical with his Iris virginica. For Iris versicolor I have studied figs. 187 and 188 of Dillenius, Tab. VI and VIII of Ehret (pl. 35), and a photograph (pl. 36, fig. 1) of Dillenius’ specimen in his herbarium at Oxford. Ehret’s [Vor. 15 248 ANNALS OF THE MISSOURI BOTANICAL GARDEN excellent figures leave no doubt about the identification. Par- ticularly to be noticed are the short ovary, the lack of auricles on the dissected style, the broad-bladed sepal, and the lanceolate petals. Dillenius' figure and his specimen show very small petals; the herbarium specimen in addition has a short ovary. In the case of Iris virginica, І have examined Clayton’s No. 259 at the British Museum, and Linnaeus’ own specimen of Jris virginica at the Linnaean Society. Both are in an excellent state of preservation and each shows heavy pubescence on the blade of the sepal. Jacquin’s plate of Iris virginica is an excellent figure of Iris versicolor. It shows a plant with small petals, sepals with no trace of a yellow eye, and long pedicels. The plate (pl. 36, fig. 2) accompanying Radius’ description of Iris carolina shows it to be identical with 1748 virginica. The sepal and petal are both waved, a character often assumed by Iris virginica, never by Iris versicolor. The petals are nearly as long as the sepals, and according to the description the plants have few flowers. Furthermore, the species is distinguished from Iris virginica of Linnaeus, according to Jacquin, which as we have seen, is 1718 versicolor of Linnaeus. Plants from the same general region were described as Iris caroliniana by Watson in 1898. In his description he speaks of “the elliptical blade lilac, veined with purple and with a yellow spot reaching to the center." The petals are described as oblong- spatulate. The type specimen, in the Gray Herbarium, shows the typical roundish seeds with a spongy surface, which character- ize Iris virginica. Material from practically the same locality was studied in the spring of 1928 by the writer. When large numbers of plants were examined there was found to be no consistent difference of any sort between the irises of the Carolina Coastal Plain and those of the Mississippi Valley. There is therefore no ground for maintaining Iris Shrevei of Small. He distinguishes it from Iris versicolor but makes no mention of its relationship to Iris carolina (Iris virginica L.), with which it is identical. The illustration accompanying his description is evidently made from a badly stunted plant. Specimens collected from the same locality visited by Small are much larger than those 1928] ANDERSON— PROBLEM OF SPECIES IN IRIS 249 in his illustration. The locality was visited when the plants were in full bloom and nearly all bore the bright yellow-pubescent spot on their sepals which shows in his illustration of Iris carolina but is lacking from his illustration of Iris Shrevei. Furthermore, plants transplanted from that locality to the experimental garden have borne as many round as D-shaped seeds, though his illustra- tion shows only a D-shaped seed. COMPARATIVE MORPHOLOGY Distinguishing differences in italics. See also pls. 37-39, 41-43. IRIS VIRGINICA L. ROOTSTALK A creeping rhizome 2.0-4.0 cm. in di- нека in fresh specimens, yellowish dirty yellow. Cortex pale pinkish white. ROOTS Wrinkled, 2-4 mm. in diam., white. LEAVES Narrowly ensiform, 1-5 cm. broad, gray-green, often flushed with purple below. STEM Sparingly branched above, green or greenish purple. SPATHE VALVES 4-12 ст. long, occasionally becoming foliaceous. PEDICELS 2.5-8.0 cm. long in the flower, rarely equalling the longest spathe valve. OVARY : 1.8-3.8 ст. long in Ше flower; ovary wall relatively AS ovules nearly filling locules at anthe. TUBE Wall of tube thick, conspicuously glandular, closely appressed to style base within. IRIS VERSICOLOR L. A creeping rhizome 1.0-8.6 cm. in di- meter. Stele, in fresh specimens, yellowish white to dirty yellow. Cortex pale pinkish white. Wrinkled, 1-3 mm. in diam., white. Narrowly ensiform, 0.5-3.0 cm. broad, gray-green, often flushed with purple below. Freely branched above, green ог greenish purple. 3-4.5 ст. long, never becoming foli- aceous. 1.0-8.0 cm. long in the flower, often ualling or surpassing the longest spathe valve. 0.8-2.0 cm. long in the flower; ovary wall relatively thin; ovules filling scarcely half the locules at anthesis. Wall of tube thin, not conspicuousl glandular, style base distinctly separate from tube. [Vor. 15 250 ANNALS OF THE MISSOURI BOTANICAL GARDEN Camera-lucida drawing of cross-sections of ovaries of Г. versicolor (right) and т. virginica (left), resin ducts in black, vascular bundles outlined; Х 10. SEPALS 4.0-8.4 ст. long, 1.6-4.0 ст. broad; blade of вера! oblong-ovate to ovate, usually some shade of bright blue, al- though white and lavender specimens have been found. Blade bearing a bright yellow conspicu- ously pubescent spot at the base. Hairs of the pubescence conspicuous and macroscopic, longer than the thick- ness of the sepal. PETALS Oblong-lanceolate to oblong-spatulate, 3.0-7.0 cm. long, 1.0-3.0 cm. broad, often (in about 50 per cent of the cases) notched at the apex, of the same gen- eral color as the sepals, delicate in texture, easily bruised, wilting readily. STYLE Style branches inwardly auriculate at their convergences. STYLE CRESTS Reflexed, variously toothed, 0.7-2.0 cm. in height. 22 a triangular tongue. STAM юан 0.9-8.1 ста. long. Pollen-grains oblong-ovate in outline. 0-7.2 cm. long, 1.8-4.0 cm. broad; blade of sepal ovate to reniform-ovate, usually some shade of purplish-blue, although white and lavender speci- mens have been found. Blade often without a conspicuous spot the base; when present, greenish or greenish yellow. Hairs of the pubescence inconspi and microscopic, shorter din the the thickness of the sepal. Lanceolate to oblong-lanceolate, 2.2- 5.4 ст. long, 0.5-2.2 cm. broad, very rarely notched at the apex, of the same general color as the sepals; firm in texture, not readily wilting. Style branches not auriculate at their convergences, or at most but weakly so. Reflexed, variously toothed, 0.7-1.6 ст. ш height. Stigma а triangular tongue. Anthers 0.7-1.6 ст. long. Pollen-grains oblong-ovate in outline. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 251 CAPSULE Capsular lining irregularly minute- Capsular lining regularly әуені striate; inner surface mainly dull. striate, giving the inner surface uniformly shiny appearance. SEEDS Round to D-shaped in outline. Always D-shaped in outline. Seed-coat corky and dull, the surface Seed-coat relatively thin, hard, and appearing irregular, with occasional shiny, the surface appearing гарману broad depressions under а hand-lens. pitted, the pits in definite rows under a hand-lens, ца Owen Sound, Ont Sreveen Creex,N. Y. № Xe La Crosse, Wia — Ho..ow Воск, Теми. Anna, luc. В.есат Len, Minn Талмкеммота, Мом, 8 Сонемтмоо, N.Y Orroro, №. М. Зтватеово, М. H. Fig. 3. Camera-lucida drawings of cells in outer seed-coat: upper row, Iris versi- color, lower row, 1. virginica; Х 20. GEOGRAPHICAL DISTRIBUTION It is planned to give a detailed discussion of the distribution of the two species in a later section. Until then certain outstand- ing points may be briefly summarized. Tris versicolor is generally a northern and eastern species, Iris virginica a southern and central-western one. As shown by the map in fig. 1, Iris versicolor occurs in New England, New York, РО оа, northern Michigan, Wisconsin, and Minnesota, and eastern Canada, 1718 virginica extends from the Gulf states to central Michigan and Wisconsin and west to the eastern edge of Nebraska and Kansas. On the Atlantic coast it is common in the Carolina Coastal plain, though it is rare in the mountains and was not seen in the Piedmont. At their zone of contact in Michigan, Wisconsin, and Minnesota, the range of Iris versicolor coincides very nearly with the northern [У от. 15 252 ANNALS OF THE MISSOURI BOTANICAL GARDEN MNA! 00.0 ) (1, Fig. 4. Camera-lucida drawings of seed capsules showing outline and сговв-вес- tion with seeds, X 14: A, I. virginica, from La Crosse, Wis.; B, I. versicolor, from Conewango, N. Y.; C, J. birgindéa, from Bonnieville, Ky. coniferous forest, that of Iris virginica with the southern hard- woods. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 253 Both species, while distinctly moisture-loving, belong to the border zone between marsh and dry land. They are not able to compete successfully if the situation is wet enough for Турћа or dry enough for grasses. In those regions which have been com- pletely drained and brought under cultivation they have very nearly disappeared. None were found in the drained areas of southeastern Missouri, though the species was formerly common there. Several local naturalists have reported that Iris virginica was much commoner in Tennessee and North Carolina before the lowlands in these states were brought under cultivation. In those sections of the country, on the other hand, where such areas have been left as pasture lands, both species have increased. Livestock eat the grass in preference to the Iris, and the iris colonies spread out vigorously, often covering several acres. The seeds of both species float easily, and seedlings can often be observed coming up on wet sandy beaches of the Great Lakes. It would not be surprising to find Jris virginica establishing itself locally along the St. Lawrence River system. Two such colonies have actually been found: one on a deserted lake beach at Saint Ignace, and one on the shores of the Niagara River a few miles above the Falls on the American side. ABBREVIATIONS The abbreviations used to indicate the herbaria in which the specimens occur are as follows: A = Academy of Natural Sciences of Philadelphia. BM = British Museum. МВС = Missouri Botanical Garden. US = United States National Herbarium. TAXONOMY Iris versicolor L. Sp. Pl. ed. 1, 39. 1753; Curtis, Bot. Mag. 1:71. 21. 1790; Redouté, Liliacées 6: pl. 339. 1812; Edwards, Bot. Gard. 2: pl. 30, fig. 2. 1812; Baker in Gard. Chron. М. S. 6:614- 615. 1876; Meehan, Native Flowers & Ferns 1: pl. 36. 1878; Millspaugh, Med. Plants 2: pl. 173. 1892; Britton & Brown, Ill. Flora, ed. 1, 1: 448, pl. 1069. 1896, in part; Small, Fl. Southeast. U. S. 306. 1903, and ed. 2,305. 1913, in part; [Vor. 15 254 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Gray’s Manual, ed. 7, 299. 1908, in part; Britton & Brown, Ш. Flora, ед. 2, 1: 537, pl. 1328. 1913, in part; Dykes, Genus Iris, 79-81. 1913, in part; Small, Addisonia 9: 55-56, pl. 316. 1924. Pls. 38, 40, 41 (left fig.); 42 fig. 2; 43 figs. 6-10. [Iris americana versicolor stylo non crenato Dillenius, Hort. El- tham. t. 155, pl. 187. 1732.] [Iris americana versicolor stylo crenato Dillenius, Hort. Eltham. t. 155, pl. 188. 1732.) [Iris latifolia Virginiana..... Ehret, Plantae Depictae pl. 6 & 8. 1748.] Iris virginica Г. асс. to Jacquin, Icones Plant. Rariorum 2: pl. 228. 1786-1793. Iris virginica L. Sp. РІ. ed. 1, 39. 1753; Michx. Fl. Bor. Amer. 22. 1803; Sims in Curtis’ Bot. Mag. 19: pl. 703. 1804; Baker in Gard. Chron. N. S. 6: 615. 1876. Pls. 34, 37, 39, 41 (right fig.); 42 figs. 1, 3, 4; 43 figs. 1-5; 44. [Iris corollis imberbibus.............. Gronovius, Flora Virginica, 11. 1739.] Iris carolina Radius, Naturforsch. Ges. Leipzig Schrift. 1: 158, pl. 3. 1822; Small, Addisonia 9: 49-50, pl. 313. 1924. Iris versicolor L. іп part; Britton & Brown, Ш. Flora, ед. 1, 1: 448, pl. 1069. 1896; Small, Е]. Southeast. U. S. 306. 1903, and ed. 2, 305. 1913; Gray’s Manual, ed 7, 299. 1908; Britton & Brown, Ill. Flora, ed. 2, 1: 537, pl. 1328. 1913; Dykes, Genus Iris, 79-81. 1913. Iris caroliniana Wats. in Gray's Manual, ed. 6, 514. 1890; Baker, Handbook Irid. 12. 1892; Sargent, Gard. & Forest 6:334- 335. 1893; Britton & Brown, Ш. Flora, ed. 1, 1: 449, pl. 1071. 1896; Wats. in Proc. Am. Acad. 25: 134. 1898; Small, Fl. Southeast. U. S. ed. 1, 306. 1903, and ed. 2, 305. 1913; Gray’s Manual, ed. 7, 300. 1908; Stapf in Curtis' Bot. Mag. ТУ, 8: 94, pl. 8465. 1912. Iris versicolor L. var. virgimea L., ex. Baker (err. typ.), Hand- book Irid. 12. 1892. Iris georgiana Britton in Britton & Brown, Ill. Flora, ed. 2, 1: 537, pl. 1830. 1913. Iris Shrevei Small, Addisonia 12: 13-14, pl. 391. 1927. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 255 IRIS VERSICOLOR L. Specimens examined: CANADA: Nova Scotia: Canso, July 24, 1901, Ward (US); Sable Is., Aug. 16, 1913, St. John 1184 (US). New Brunswick: Campobello Is., July 8, 1880, Smith (US). Ontario: Lincoln Co., June 10, 1897, McCalla (US); Paris, vicinity, July 4, 1926, Mathias 636 (MBG). UNITED STATES: Marine: St. John River, July 22, 1917, St. John & Nichols 2280 (US); Westbrook, June 8, 1906, Ricker 50 (US); Fort Kent, river flats, July 10, 1908, Mackenzie 3422 (MBG); Seal Harbor, swamp, June 28, 1887, Redfield 7117 (MBG); St. Francis, gravelly shore of St. John River, Aug. 5, 1893, Fernald 147 (MBG). New НАМРЗНТВЕ: Peterborough, Aug. 20, 1913, Batchelder (ASP); Shelburne, June 17, 1915, Deane (US); Shelburne, Oct. 28, 1915, Deane (US); Connecticut Lakes, Oct. 20, 1895, Stevens (US). Укнмомт: Brandon, swale, June 6, 1921, Dutton (MBG); So. Burlington, July 8, 1914, Knowlton (ASP). MassaAcHusETTS: Dedham, June 13, 1897, Greenman 2333 (MBG); Brookline, June 26, 1896, Greenman 2321 (MBG); Granville, meadow brook, June 20, 1914, Seymour 174 (MBG); So. Framingham, May 31, 1890, Sturtevant (MBG); Cambridge, May 29,1901, Floyd (MBG); Nantucket, pond margin, July, 1923, Mason (MBG); Ashland, wet grounds, Morong (MBG); Nonquit, June 5, 1888, Sturtevant (MBG); Woods Hole, July 26, 1911, Pennell 3164 (ASP); Wilbraham, June 12, 1876, Pillsbury (ASP); Boston, 1816, Boott (US); Marthas Vineyard, June 29, 1916, Seymour 6158 (US); Nantucket, June 2, 1900, Day 64 (US); Marthas Vineyard, Aug. 1888, Harrison (US). Connecticut: Middletown, May, 1836, Bigelow (MBG); exact locality and date lacking, Wright (MBG); Stratford, June 7, 1893, Eames (US). New York: Otis Summit, July 27, 1903, Williamson (ASP); St. Regis Falls, June 28, 1903, Hudson 31 (US); Ithaca, June 23, 1885, Coville (US); Van Courtlandt, June, 1893, Pollard (US); N. Harpersfield, June 21, 1906, Topping (US) ; Elizabethtown, swamps, [Vor. 15 256 ANNALS OF THE MISSOURI BOTANICAL GARDEN July 2, 1875, Redfield 7911 (MBG); Ithaca, June 4, 1877, Trelease (MBG); east of Cherry Valley, July 2, 1926, Mathias 652 (MBG); banks of Lake George, Aug. 24, 1856, Engelmann (MBG); locality lacking, 1840, Short (MBG); Bedminster, meadows, date lacking, Perry (MBG). New Jersey: Vineland, Sept. 12, 1923, Bassett & Long (ASP); Camden, May 22, 1922, Bassett (ASP); Quaker Bridge, Sept. 11, 1923, Dreisbach 1834 (ASP); Folsom, June 10, 1911, Long 5940 (ASP); Cor’s, Ocean Co., Oct. 11, 1910, Long 5582 (ASP); Cold Spring, Cape May Co., June 1, 1911, Brown (ASP); Atlantic City, margin of meadows, June 3, 1875, Redfield 7910 (MBG). PENNSYLVANIA: Lester, Delaware Co., May 28, 1912, Findlay (ASP); Chester Co., June 4, 1905, Weston 164 (US); Harris- burg, Aug. 13, 1888, Small (US). MARYLAND: Spesutic Is., May 25, 1879, Smith (US) ; Chester- town, Aug. 18, 1900, Vanata (ASP). District or Согомвта: Washington, May and June, 1878, Ward (US). MicnuiGAN: Cheboygan Co., sedge pool, July 17, 1917, Gates & Gates 10597 (MBG); Douglas Lake, Cheboygan Co., July 17, 1916, Ehlers (MBG); Burt Lake, Cheboygan Co., July 4, 1917, Gates & Gates 10492 (MBG); L’Anse Co., July, 1892, Eby (MBG); Keweenaw Peninsula, Aug. 1878, Engelmann (MBG). Wisconsin: La Pointe, Sept. 3, 1878, Engelmann (MBG). IRIS VIRGINICA L. Specimens examined: UNITED STATES: District oF Сотомв1А: Alexander Is., May 15, 1915, Van Eseltine 366 (US). VinaiNIA: Norfolk Co., Мау 11, 1898, Kearney 1079 (US); exact locality and date lacking, Clayton 259 (BM). Ховтн САБОШМА: Hendersonville, wet meadows, May 26, 1898, Biltmore Herb. 542b (MBG, US); Roandale Farm, June 8, 1895, Wetherby 1067 (US). SouTH CAROLINA: Cat Island, Georgetown Co., Aug. 16, 1915, Alexander 111 (US, NY). Ошо: Middleburg, Cuyahoga Co., June 20, 1897, Watson 1928] ANDERSON— PROBLEM OF SPECIES IN IRIS 257 (MBG); Clare, Hamilton Co., spring-fed swamp, Мау 30, 1906, Braun (MBG). Kentucky: Elizabethtown, May 23, 1926, Anderson Ф Woodson 38 (MBG); exact locality and date lacking, Shor (МВО); Bowling Green, swamp, May 21, 1897, Price (MBG); Drake’s Creek, Warren Co., May, 1891, Price (MBG); Livermore, Mc- Lean Co., June 2, 1920, Е. J. Palmer 17707 (MBG); Ohio City, swamp, June 5, 1901, Price (MBG); Clay City, May 23, 1908, Biltmore Herb. 542 (US). TENNESSEE: Stanton, May 20, 1926, Anderson & Woodson 60 (MBG); Knoxville, wet ground, May 1896, Ruth (MBG). GEORGIA: McQueen Is, Chatham Co., brackish marshes, April 30, 1904, Harper 2180 (MBG). Fromm: Lake City, Мау 23, 1897, MacKenzie (MBG). МїснївАМ: Adrian, June 24, 1901, Dewey 508 (US); Ann Arbor, June 10, 1884, Sudworth 50 (US); Flint, swamp, June 18, 1923, Anderson (MBG). INDIANA: Terre Haute, Мау 19, 1889, Evermann (US); Roby, June 18, 1910, Lansing 2789 (US); Miller’s, May 30, 1902, Chase 1799 (US). ALABAMA: White River marshes, April 14, 1898, Mohr (US); Hollywood, May 15, 1902, Biltmore Herb. 5420 (US); Pearlington, April 8, 1880, Mohr (US) ; Mobile, Мау 16, 1893, Mohr (US). Пллхотв: Spoon River, Stark Co., June 9, 1907, Chase 1425 (US); Macon Co., June 12, 1915, Clokey 2433 (US); Morgan Park Ridge, May 27, 1907, Dixon & Gage 702 (US); Kankakee, May 27, 1913, Crampton 124 (US); Decatur Co., May 29, year lacking, Clokey (MBG); French Village, St. Clair Co., May 27, 1903, Eggert (MBG); Olney, April 14, 1921, Ridgway 1381 (МВС); Fish Lake, St. Clair Co., July 16, 1858, Norton (MBG); Wood- lawn, Jefferson Co., May 16, 1898, Eggert (MBG); Decatur, date lacking, Clokey 2789 (MBG); Shawneetown, swampy borders of lake, June 19, 1919, Е. J. Palmer 15554 (MBG); Reevesville, Johnson Co., low swampy open ground, June 3, 1919, Е. J. Palmer 15350 (MBG); Mounds, Pulaski Co., May 7, 1919, Е. J. Palmer 15074 (MBG). Мїв81881РРГ: Beauvoir, March 28, 1898, Tracy 4463 (MBG). Minnesota: Nicollet, June, 1892, Ballard (US); Swan Lake, [Vor. 15 258 ANNALS OF THE MISSOURI BOTANICAL GARDEN July 16, 1917, Metcalf 40 (US); Fort Snelling, June 22, 1888, For- wood (US). Iowa: Tiffin, June 5, 1909, Somes 3132 (US); Fayette Co., June 5, 1894, Fink 37 (US); Hamilton Co., July 1891, Rolfs (MBG); Missouri Valley, June 21, 1897, Pammel 587 (MBG); Iowa City, date lacking, Hitchcock (MBG); Armstrong, Emmet Co., June 18, 1897, Cratty (MBG); Ames, June 21, 1897, Ball & Meeker 524 (MBG). Missourt: St. Louis Co., June 1912, Craig (MBG); Oakwood, Ralls Co., open woods, June 19, 1917, Davis 7553 (MBG); Randolph, May 18, 1895, Mackenzie 544 (МВС); Jackson Со., low wet ground, common, June 17, 1892, Bush 1475 (MBG); Jefferson City, June 1870, Krause (MBG); Carthage, swamps, May 27, 1906, Е. J. Palmer 883 (MBG); Kimmswick, May 23, 1885, Wislizenus 407 (MBG) ; Winfield, Lincoln Co., June 7, 1916, Davis 1456 (MBG). ARKANSAS: Nettleton, May 7, 1893, Eggert (MBG); Craig- head, lakes, May 7, 1893, Eggert (MBG). Louisiana: Hammond, April 4, 1889, Gallup 16 (US); Minden, April 14, 1901, Trelease (MBG); Madisonville, May 5, 1888, Joor (МВС); Covington, April 1920, Arsene 12400 (US). NzBRASKA: Nebraska City, June, 1889, Webber (МВС). Kansas: Wyandotte, moist places, Aug. 5, 1897, Clothier 1067 (MBG, US). Texas: Troupe, Мау 9, 1902, Reverchon 2793 (MBG, US); Orange, April 19, 1899, Bray 61 (US); Keechi, Leon Co., April 20, 1918, Е. J. Palmer 13420 (MBG); Beaumont, Jefferson Со., April 22, 1916, Е. J. Palmer 9522 (МВС). ПІ. INTRA-SPECIFIC VARIATION IN IRIS VERSICOLOR AND IRIS VIRGINICA Wherever either species was studied, the individual plants which went to make up a colony were found to vary strikingly among themselves. They varied in every conceivable character- istic, both vegetative and floral, though the latter variations wer the most conspicuous. The flowers varied in size and form, in color and color pattern, in number and arrangement, in texture, and in odor. Plate 34 shows such of these differences as can be ARD., Vor. 15, 1928 1 1: Амм. Мо. Вот. ( Three flowers each of six plants of Iris virginica from Portage des Sioux, Mo. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 259 recorded by the camera for six individuals from a typical colony, that of Iris virginica at Portage des Sioux, Missouri. They are not selected extremes but are the first six individuals which were located with at least three flowers apiece. The pictures were taken under uniform conditions as regards lighting, exposures, development, etc., and such differences as occur are due to differences in the color and texture of the flowers themselves. It may not be out of place to call attention to some of the differences which obtain between these six plants, since they are similar to the differences which were met with everywhere throughout the study. Plant No. 1. Petals narrow and ruffled, ovary short. Plant No. 2. Petals flat and wide, flower pale blue in color. Plant No. 3. Entire plant large and robust, of great vegeta- tive vigor, clone covering half an acre, flowers marked with dark blue veins on a light back- ground, spot on sepal large and brilliant. Plant No. 4. Flowers small and light colored, sepals broadly spatulate. Plant No. 5. Flowers pale gray-blue, sepals long and narrow, stems and leaves very short, plant floriferous. Plant No. 6. Flowers dark reddish blue, ovary short, stems slender, few-flowered. In order to summarize and average such differences as these it becomes necessary to select a few for concentrated study. The characters chosen should fulfill three conditions; they should be easily measurable; they should show some variability; and they should not be easily affected by environmental influences. The first qualification eliminates a number of very conspicuous differences, such as color and color pattern, which are not adapted to quick and accurate measurement. The second bars those characters, such as the length of the anther, which are practically indentical for a large number of species of Jris. All of the characteristics of the plant are, of course, open to the objection that they are affected by the environment but some of them are much more stable than others. The leaf characters are particu- larly poor. The largest leaf on a plant may be two or three times [Vor. 15 260 ANNALS OF THE MISSOURI BOTANICAL GARDEN as wide as the smallest. The size and shape of the seed capsules are likewise extremely variable on the same plant in different seasons and under different conditions. In general the dimen- sions of the floral elements are less easily affected, and since they are of predominant importance from a taxonomic standpoint they were very largely used in the present work. After a year’s preliminary study, seven characters were chosen for measurement and have been used throughout the work. Five others were subsequently tried extensively, and three of these are being used at the present time. The characters chosen and the exact manner in which measurements were taken are shown in fig. 5 These characters are measured as follows: Sepal length. This is the maximum length of the sepal from the base of the stamen to the tip. Sepal width. The maximum width at right angles to the length. Sepal taper. Distance from the tip to the point of maximum width. Petal length. Unfortunately there was no exact lower boundary as in the case of the sepal, and some of the variation in the earlier measurements is due to the fact that different points were used as а base. It was eventually defined as the line between the bases of the lateral flanges at which it breaks off naturally when pulled over backward. Petal width. The maximum width at right angles to the length. Petaltaper. The distance from the tip to the point of maximum width. Crest. The maximum length of the appendages of the stigma, measured from their tips to а line connecting the ends of the stigmatic lip. Ovary. 'The junction of the ovary and pedicel is distinct in both species; that between the ovary and the tube is much less so, being marked by a vague ridge. 'The summit of this ridge is taken as the line of demarcation and the ovary is measured from it to the base. Tube. The tube is measured from the same point to the base of the stamen, one sepal having been removed. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 261 Stamen. The length of the anther from the attachment of the filament to the tip. Care was taken to measure only fully opened, uninjured flowers. Not more than one flower of a clone was measured (except in those few cases where intra-clonal variation was being studied). It was found upon experiment that one single set of measurements gave almost as consistent results as when two sepals and two petals were measured on each plant. An experiment was made on the Fig. 5. Diagram showing how floral elements of Iris were measured. colony at Portage des Sioux, Missouri, to determine if the measure- ments were being affected by the flowers not being fully expanded. Twenty flowers were measured at four o’clock in the afternoon and remeasured at seven the next morning with practically identical results. The first five year’s measurements have been summarized in tables 1-1v. The figures in italics are the class containing the median or mid value. The median has been used as an average rather than the mathematical mean, since it is less influenced by occasional extreme values. 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ДЕДЕ 81412 ono uon [6261 т|т|Р7|018|9 I 8 5194" очо 349919) ШИ 8261 eee $ с'ејо $ йо оо 2 а е је ... "v PVT oo Атап |7261 че РЕЛЕ змо] Ima 94) FZ6T ЭЭ 11341215 змо] ята |#с6Т "lzisiolt?*t[IIll S змој| товтрву 310 261 e аа ҒА” uuo 9917 ДОМ 8261 трегегіІІЗ 111819 1 ок 9[[IASÁw И 916 Тр Tie Te T|T T 8' I4 I9 TI I ест влодошт ово ш 43813] 194}UV 91835 uxor, (рәпапаоо) VOINIOSIA SII 278 (penuguo)) AI ЧЛАУЈ, Жэ Dn 1928] 279 ANDERSON—PROBLEM OF SPECIES IN IRIS зчор od 91339 |2261 y ET га e "E эло ‘SI 29194 |9261 ££ 2|16|:2|%491Т1|1 Мж ‘SIM зевицв 7261 95, ААИ САК КЕЛЕ МЕ МЕ Е ВАС 2 И АДК Бай КЕЕ АКИ SUM oqmoopreq 7261 IG 2|8181112101 61816 “SIM әвволгу 8"11Р261 от ^H Lb 316: DLE иш ртвАХЗН Р 61 y y re ES) БЕБЕ uur HəqAIy|pZ6I е? ТІТ|8ІІЦ7Д91|6 WA човолепи робт “, • © ... өте ... "Чи роомлгт 1661 02 8|(115141013 ! 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T9 16 ТУ ТЕ ТСТТ ТОТ Ве ВЕ ма - 6 ~ влодошт идо UI 45910) dan 819jeurquoo ш чу че чүчү 97815 чир], 5 ч 9715 о) З0итріоәәв pogisse[o Апојоо цово јо врепртатриј (рәпптаоо) VOINIDUIA БІЗІ (penuguo)) ДІ ЧЛаУЈ, Vor. 15 са ч ў кеа . поз: и eo чн үч : «00 і әз > © - 5 Б a E E Ф ж e са e — -= NNN NN er сб сб =N |8имо4 8261 1 @Ф ч 0 ду = © ~ “000 ‘носо ET i + 100 = само са H ср оо са ны = со БЭЭ — BOO 00i “CORED iQ DM с, (OQ r.i men . ра — N Що = © © PONN OO un nc OND “ооо оок со ~ $ ч сз — тч 1оодлол 7 1 |/261 “1111 1 11 сени 141 14 194 14 1940-1010 1 00 роон |7261 9 те тр те те тт 1016 |8' 12 0:06 1814 19 18 ТР 16 Tjo TT 1016 8 4 $19]9UIT)U99 UI 95910) $19j9urruoo ит 4383] лэчзиу әуеде пмој, вх. 9218 0} Зитраодов pogissw[o Kuo[oo цове јо втвпргАтри ANNALS OF THE MISSOURI BOTANICAL GARDEN ЧОТООІБЧЯА SII 280 (рапициод) ДІ ATAVL 1928] 281 PROBLEM OF SPECIES IN IRIS ANDERSON 02 Те | г | # |о | ? | < - 110 чорпо 17261 РІ (S Meg ea: 1» ВЕЖЕ: — ; t| то 918Х “М|Рб61 82. (82 11111111 — чети оштрваи 1061 се с | та | 8 | 412 189! - чети eoeusp 3S 7661 11 а "T - чети ввмв T, “11061 11 bu uisus е [eee РОС rae е + е I ова [| с = а» | » = +» Есен Чә Арто SEMU 1 2761 Е 3026 18 12 19 те тр те те тт 1016 | 12: |2 20 с6 18 та 19 те тр те те цг тот’ во ве Bg на к = 5193э 999 UI 48910) За S19jeurruoo ur 438343] 1эЧз чу 97835 UAOT, 8 ~ 9218 о) Surpiooo€ poegisse[o Áuo[oo цове јо spenptarpug VOINIDOUIA “IX чотоотвнял SINI (рәпацао)) ДІ Я1ЯУ1. Мог. 15 282 ANNALS OF THE MISSOURI BOTANICAL GARDEN separating the two species. It is thus apparent at the outset that no biometric method of distinguishing the two species can be a simple matter. While it is certainly true, as Hall and Clements (728) and McLeod (726) have suggested, that taxonomy needs the development of more exact methods, there are serious limitations to a wholesale inclusion of biometry in every-day taxonomic procedure. These limitations may be easily demonstrated by a simple example. Figure 6 shows five petals of 1748 virginica and five ибей ІШІК Fig. 6. А, outlines of five petals each of Г. versicolor; B, outlines of five petals each of I. virginica, of Iris versicolor. For the purpose of taxonomic description they may be conveniently and accurately separated by describing those of Iris versicolor as ovate-lanceolate and those of Iris virginica as И дэ уа чачу ан" 30е Ааа 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 283 obovate-spatulate. To separate them by biometric methods is by no means so easy, as table v shows. TABLE V Petal Petal Petal Taper length width taper width Iris virginica cm. cm. еш. №. $ 4.2 1.4 1.5 1.1 2 4.7 ТЕТ 0.8 0.5 8 4.8 157 0.9 0.6 4 4.6 1.8 1.1 0.9 5 4.6 1.6 1.2 0.8 Iris versicolor No. 1 4.7 1.3 2.0 1.5 2 4.8 1.5 125. 1.1 3 4.1 132 1.8 1.3 4 3.0 1.2 1.2 1.0 5 4.5 1.4 27 1.2 Neither thelength nor Ше width willsuffice. The taper (1. e. the length between the point of maximum width and the tip) is only somewhat better. The ratio between this latter measurement and the width is still better though it fails to separate the entire lot. None of the measurements is as good for purposes of distinc- tion as the single terms “оуже-апсео]а4е” and ‘‘obovate- spatulate." Only by combining all three measurements into & complex ratio would it be possible to demonstrate, mathemat- ically, the discontinuity between the two sets of petals. As Minot (quoted by Thompson, 717) has said, “The fact that men of genius have evolved wonderful methods of dealing with numerical relations should not blind us to another fact, namely, that the observational basis of mathematics is, psychologically speaking, very minute compared with the observational basis of even a single minor branch of biology. * * * While therefore here and there the mathematical methods may aid us, we need а kind and degree of accuracy of which mathematics is absolutely incapable. ” Тће above example illustrates the two chief reasons why biom- etry must necessarily be limited in its application to taxonomy. In the first place, mathematics is swift and efficient only in re- cording differences in number; it becomes cumbersome in record- [Vor. 15 284 ANNALS OF THE MISSOURI BOTANICAL GARDEN ing differences in form, however useful it may be for a deeper analysis of the forces which produced the form. Yet it is just such differences in form which are most commonly met with in taxonomic work. This point is shown in a survey of the points on which specific distinction is based in two representative families. (one from the Monocotyledons and one from the Dicotyledons) in the seventh edition of Gray’s ‘Manual’. The differences were classified as being based on shape, on absolute size and number, and on comparative size and number. In all doubtful cases the preference was given to differences in number. А А Differences in Differences in pem 2” - absolute size comparative size р ог number Tridaceae 21 13 12 Boraginaceae 31 11 9 There is an even more fundamental reason why the customary methods of mathematics are not well adapted to taxonomic work. Such methods are comparatively simple when the variations in one characteristic are being traced; they become involved and cumbersome when the simultaneous changes in a large number of variables are studied. Yet this last is essentially the method of the taxonomist. When he distinguishes between a group of sugar maples and a group of silver maples, for instance, he is summarizing a large number of differences—differences in form, size, color, and color pattern. Mathematics can deal with such problems through the study of correlation, but it is slow and laborious work and though it may be useful in the analysis of some particular problem it is not adapted to general taxonomic use. An attempt has therefore been made to develop a new method of presenting biometric data which would combine the good points of the methods of mathematics and comparative morphol- ogy. Like mathematics, it is accurate and objective. Like morphology, it leaves something to the trained eye. This has been accomplished by diagramming the data in a series of ideographs. Figures 7 and 8 show how the four measurements on the petal and sepal are combined into a single figure. Essen- 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 285 и Fig. 8. Diagram showing typical flower of J. versicolor and resulting ideograph. tially the ideograph consists of a diagrammatic white petal, superimposed upon a diagrammatic black sepal. By construct- ing an ideograph of this sort for each plant measured, it is possible [Vor. 15 286 ANNALS OF THE MISSOURI BOTANICAL GARDEN to show simultaneously the variation in the four variables con- sidered, for an entire population of plants, or to compare the variation in one population with that in another as in figs. 10 to13. Such ideographs would seem to be of general usefulness in taxonomic work. They are capable of demonstrating slight differences in proportion which are not revealed by figures alone. In the genus Iris, for instance, though Iris fulva, Iris foetidissima, and Iris prismatica each belong to a different section of the genus, the three species have flowers of nearly the same size. The dimensions of the petals and sepals are so nearly the same that the species hardly appear distinct when the figures are compared, as in table ут. When these are arranged as ideographs, however, as in fig. 9, the essential differences in proportion between the three species are clearly demonstrated and they are seen to be morphologically distinct. TABLE VI COMPARISON OF FLORAL DIMENSIONS OF THREE SPECIES OF IRIS Sepal length | Sepal width | Petal length | Petal width іп em. in cm. іп em. in cm. 4.4 2.0 4.0 1.1 4.9 1.8 4.3 12 Iris prismatica 4.7 2.1 4.3 1.4 4.8 1.8 4.6 1:1 4.5 1.8 4.2 1:1 4,9 2:% 3.7 1.4 4.9 2.4 4.0 1:2 Tris fulva 5.6 2.6 4.5 1.6 5.3 2.4 4.1 1.3 5.0 2.2 3.9 1,1 4.8 1.8 3.9 0.9 3.9 1.6 3.6 0.8 Iris foetidissima 4.0 1.8 3.6 0.9 4.4 2.0 3.9 0.8 4.5 21] 4.0 0.9 Ideographs for twenty individuals each of sixteen repre- sentative colonies are grouped in figs. 10-12 ([. virginica) and fig. 13 (I. versicolor). They give another proof of the strik- ing variation in size and proportion which has been found in every colony studied. In marked contrast to the variation be- tween individuals is the general resemblance between colonies of the same species. While several different colonies have slight 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 287 = J ПИ ПП ІШ газа Fig. 9. А, ideographs of five plants of Г. fulva; В rn of five plants of J. foetidissima; C, ideographs of five plants of 1. prismat individual tendencies, and while in the case of those colonies which have been measured in successive years (fig. 10) the peculiarities persist from year to year, there is practically no differentiation between one region and another. The only generalization that can be made is that Iris versicolor becomes on the average a little smaller as one goes from north to south and that Iris virginica becomes a little larger. "Thus, although the colony at Stanton, Kentucky (fig. 12), seems slightly unusual by reason of its com- paratively large petals the other colonies studied in Kentucky and Tennessee (ВопшеуШе and Camden) do not show these peculiar- ities. There is as much variation in proportion and almost as 288 ANNALS OF THE MISSOURI BOTANICAL GARDEN ППППП "mnn ППППП nnnnn ЇПППП nnnnn ППППП nnnnn nnmnnn nnnnn ШІНШІ nnnnn nnnm Anann ПП ANAAN ANNAN ППППП ППППП ППППП ППАПП ПАЛА nnnnn Sunbury, Ohio ШІ Anann ППППП ЇПППП nmn шиш ГУ от. 15 ІШІП mnn ППППП 1 ШИЛ nnnnn ПЛППП nnnnn ANANT ШЕШІ ШІ ШІІШІШІ nmn n Jackson, Miss. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 291 ШІ ПППИЛ NAAN nnnnn ANANT nnnnn nnnnn Timagami, Ont. Truro, М. В. ПААПА ПАЛАП ППППП nnnnn ППППП anann ШИ Ottawa, Ont. Hubbardsville, N. Y. Fig. 13. Ideographs of twenty individuals each of four colonies of J. versicolor. Мог. 15 292 ANNALS OF THE MISSOURI BOTANICAL GARDEN much in size in the colonies from southern Michigan as there is within the whole group of colonies of Iris virginica from the Great Lakes to the Gulf of Mexico. Above all, when the ideographs are considered as a whole, the two species remain completely and absolutely distinct. In spite of a wide range of variation in separate characteristics, when the combination as a whole is studied it is found to be strikingly TABLE VII MEDIAN MEASUREMENTS OF IRIS FLOWERS FROM DIFFERENT LOCALITIES IRIS VIRGINICA Қозы» Median| Median} Median| Median атай : вера! | вера! | petal | petal of Тар 4- Locality Year length | width | length | width in em. | in em. | in em. | in cm. 25 Kimborough, Ala. 1927 .8 .8 5.7 Ч - Jackso ‚ Miss. 1927 .9 .6 4.9 : 50 Camden, Tenn 1926 n7 .0 5.7 2.( ) Bonnieville, Ky. 1926 Т 4 5.7 : Stanton, Ky. 1926 .5 .6 4.9 Р ) Eastover, 5 1928 7 .8 5.7 à › Maysville, N. С 1928 .5 .0 6. 2. 38 ппа, Ш. 1926 6.3 + 4. 5 27 Vulcan, Ш. 1925 6.3 .6 5. А Vulcan, Ш. 1926 6.3 .6 5. : : Farmington, Ark. 1925 6.3 4 5. 2. 4 icks, Мо. 1925 5.9 . 4. 2 Valley Pa rk, Mo. 1926 5.9 А 5. ! ‹ Portage des Sioux, Мо. 1926 5.9 | 4.9 А Portage des Sioux, Мо. 1927 6.3 : 5.7 2. Portage des Sioux, Mo. 1928 8 я 5.8 2. гуа лы: Мой. 1924 ‚8 ; 5.3 Fort Madison, Та. 1924 9 : 4.5 К Sunbury, 0. 1927 24 | 5.3 л Mill Creek, О. 1925 А 4. E , 0. 1925 5 .4 4. E Bay Bridge, O 1925 T 4. E ce, Mic 1926 ) E. 4.9 2.( Schoolcraft, Mich. 1926 5.9 .( 4.9 л enterville, Mich. 1926 3 К. 5.8 y. Hartland, Mich. 1926 9 1 4.9 $. А. ‚ Mich. 1924 | 55 ~ 4. 4 4 а ich. 1924 5.5 4 4. : Otisville, Mich. 1924 5.5 jd 4. Е tisville, Mich. 1926 ) к 4. | Frankenmuth, Mich 1926 5.9 .€ 4. 12 Frankenmuth, Mich 1927 5.9 <. 4. : ) nwood, Mich. 1926 5.9 А. 4. 2. ) Linwood, Mich. 1927 5.9 .0 4. | ) Muskegon, Mich. 1924 { 4 4. 4 La Crosse, Wis. 1924 5.9 26 4. : Pardeeville, Wis. 1927 5.5 .8 4. 3 Slinger, Wis. 1924 5.9 4 4.9 1928] ANDERSON—PROBLEM OF SPECIES ГМ IRIS 293 IRIS VERSICOLOR и а Median| Median| Median| Median Aegre: А вера! | вера! | petal | petal ades неща нашу Year | length | width | length | width іп em. | in em. | in em. | in em. 28 ,M 1927 5.9 2.8 3.7 1.2 22 Liverpool, Pa 1927 5.9 3.0 4.1 .4 35 Harmonsburg, Pa 1925 5.5 2.8 3.3 |.2 29 Conewango, N 1924 5.9 3.2 4.1 .8 28 Villenova, N. 1927 5.9 3. 4.1 .6 35 |Hubbardsville, N. Y 1925 5. 2.6 3.3 .0 27 Pownal, Vt. 1925 5. 2. 3.3 .0 38 Қы Center, Vt. 1925 5. 2.: 3.3 3 87 Clarendon, Vt. 1925 5. 2. 3.3 .0 34 New Haven Jct., Vt 1926 5.9 3.( 4.1 .5 25 Middlebury, Vt 1926 5.9 8. 4.1 .4 20 игу, М 1927 5.5 2. 3.7 .4 26 Alberton, Ont 1927 5.5 8. 4.1 1.6 82 Обама, О 1926 5.9 3.( 4.1 |.4 26 Timagami, Ont 1926 5.9 8. 4.1 1.2 22 % ‚ М.Б 1927 6.: 2. 4.1 1.4 constant. Iris versicolor remains always and алатау Iris versicolor. and Iris virginica remains always and unmistakably Iris virginica. There is not the slightest tendency for one species to merge into the other. By employing another mathematical concept, that of the average, we can construct a different sort of ideograph and pro- duce something like а composite picture of each colony. If for each colony we take the average length of sepal, average width of sepal, average length of petal, and average width of petal, and construct an ideograph we will obtain a figure which will present graphically the averages of all four measurements for the colony in question. It will be a purely hypothetical figure; it will not necessarily present the kind of proportions most commonly met with in the particular colony but it will serve for convenient comparison between separate colonies and between successive measurements of the same colony. As in the earlier presentation of averages, the median is used rather than the mathematical mean because it is less influenced by occasional extreme values. The median values and also the number of individuals measured are presented in table уп. The resulting ideographs are shown in ће. 14. The examination of these composite ideographs strengthens the conclusions already arrived at. The differences between colonies ПП ПП Ш ПП Ш улова (УТО vary loa болт Аза Seanvay Ман Casa Mons Тар фачарлее Селт: Не Алмаға Yur Otssense-20 HUNDI Orontes Ғадакив теті» ПП „(а Сыма » Composite ideographs of 39 colonies of I. virginica. балама мәй? Lissus — Len were ty | Талғантыла Заөөал [Vor. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN AMALIA Hoos „Ма Livenroor a Напон: АД Сочемлисо, ЖҮ Visenova, ИХ ПЛЛЛП Huasarvsviz, NY. Бреша Bwa бей VE CLAREAPON М. Minas Bohri ППППП Моравом Оозвовх И Asaraten, Cue Отгата Dec — TinacAmi, Онч, “аваг, М 5, Composite ideographs of 16 colonies of I. versicolor. Fig. 14. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 295 are very slight. In spite of the fact that the colonies measured extend from the Gulf Coast to the Great Lakes in the case of Iris virginica, and from Maryland to the north woods in the case of Iris versicolor, there is practically no evidence: of regional differentia- tion; that is, of the formation of morphologically distinct geo- graphical subspecies within either Iris versicolor or [ris virginica. As before, Iris versicolor is seen to become slightly dwarfed to- wards the South, whereas Iris virginica is smaller in the North. The general proportions of each species, however, remain strik- ingly similar throughout their entire ranges. There is no out- standing difference in proportion which characterizes any one region, though individual colonies may show slight peculiarities. Thus while the colony at Wicks, Missouri, is distinctive by reason of its proportionately small petals, that at Valley Park, Missouri, only a few miles away, is characterized by proportionately large ones, while the near-by colonies of Portage des Sioux, Missouri, and Vulcan, Illinois, have petals of about average proportions. In addition to differences in size and proportion, another sort of difference has been noted in the planis grown in the experi- mental garden. There is a general tendency, in both species, for the plants from northern colonies to come into bloom sooner than those from southern colonies. There is a good deal of variation in blooming time within each locality, of course, and this will mask the general tendency to a certain extent but for three springs it has been observed that the earliest plants to come into bloom are from northern colonies and the latest are those from southern ones. Aside from differences in size and in blooming periods no consistent regional differences have been found within either species. It is, of course, possible that such differences existed but were not revealed by the methods used. However, these methods did distinguish successfully between the two species. If regional differences do exist, they must be of an entirely different order of value from the differences between species. CHARACTERISTIC TENDENCIES Quite as remarkable as any clear-cut differences between the two species were certain characteristic tendencies of one or the [Vor. 15 296 ANNALS OF THE MISSOURI BOTANICAL GARDEN other species to depart from the normal. The spathe valves of the two species are usually very similar, the longest ones of 1748 versicolor and the shortest of Iris virginica being quite indistin- A B Fig. 15. A, range of variation in spathe valves of Г. versicolor, Х 15; B, range of variation in spathe valves of Г. virginica, X 15. guishable morphologically. Those of Iris virginica, however, occasionally become long and foliaceous like those of Iris hexagona; those of Iris versicolor were never seen to do so. Iris virginica often (in nearly 50 per cent of the cases examined) had a terminal notch in the petal. Only twice have such notches been seen in J. versicolor. They were not due to mutilation, since they could be found in buds several days before opening; they were probably correlated with a dichotomy of the vascular system of the petal. More or less associated was a tendency on the part of Iris virginica to produce ruffled, crimped, or even crenate petals. Flowers exhibiting these characteristics are shown in pl. 34. Only once or twice was the tendency observed in /ris versicolor and even 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 297 then it was not highly developed. These are only a few of a host of variable characters which were occasionally expressed in one species and rarely or never in the other. There is nothing new in this experience. It has been the common lot of all who have attempted to distinguish groups of organisms, be they species or families or phyla. The quotation from Bergson which Mrs. Arber (725) uses in distinguishing between the Monocotyledons and the Dicotyledons is quite as appropriately used here as there. “The group must not be defined by the possession of certain characters but by its tendency to emphasize them. ” The point in calling attention to this commonplace phenomenon is to suggest that if most of the differences between species are of this sort, taxonomists might well undertake surveys of variation within the species they are treating, on a scale much larger than is eustomary at the present time. Such studies should be particu- larly important in any attempt at a phylogenetic treatment of related species. ris virginica is closely related to Iris hexagona, for instance, and one of the morphologieal indications of this relationship is that in about one case out of fifteen or twenty the spathe valves of Iris virginica are foliaceous like those of 7. hexa- gona. PECULIAR FORMS A few of the individuals observed have been so unusual as to deserve special mention. Albinos.—Complete albinos with no trace of color other than yellow have been found only in 1. virginica. These were found at two places, at Lawrence, in western Michigan, and near Lake Saint Clair, in Ontario. Partial albinos are fairly common in both species. ‘They are white, shaded or faintly lined with blue. They have been observed at the following localities: I. virginica—Lawrence, Michigan (in sarae colony with albino); Farmington, Arkansas; Maysville, S. C.; Dennison, Mich. I. versicolor—Timagami, Ontario; Central Connecticut (M. E. ains). Occasional individuals were found in each species in which the flower color is lavender or red-purple instead of blue. One such Vor. 15 298 ANNALS OF THE MISSOURI BOTANICAL GARDEN form has been offered in flower catalogues for some years under the name of J. versicolor kermisina. Similar color forms have been found at the following localities: I. virginica—Otisville, Mich.; Burke, lowa; Camden, Tenn. I. versicolor—Stratford, N. H.; Hubbardsville, N. Y. Occasional individuals were found with sepals strongly re- flexed. Three flowers from such an individual are illustrated in pl. 39, figs. 7-9. THE GENETIC RELATIONSHIPS OF INDIVIDUAL DIFFERENCES IN IRIS VERSICOLOR An experiment is under way to determine, for each species, the extent to which the peculiarities which characterize individual plants are passed on to their offspring, under the conditions obtaining in nature; in other words, to determine, from an inspection of the variation between sister seedlings, the amount of interbreeding which takes place under natural conditions. If the species is naturally self-fertilized ninety-nine times out of a hundred, all seedlings grown from a single parent plant should be practically alike and similar to the parent. If, however, there has been continuous out-crossing with other individuals, the sister seedlings should differ from each other and from the parents; as much, for instance, as human brothers and sisters differ from each other. Several such series of sister seedlings from seed-pods collected on plants of Iris versicolor at Connecticut Lakes, N. H., bloomed іп 1928. There were not enough flowers рег plant to make careful qualitative studies but there were sufficient to allow certain preliminary conclusions to be drawn. One of these series is illustrated in pl. 40, and shows one flower from each of five sister plants. It will be noted that while the flowers are in general very similar, they are not as much alike as flowers from the same plant (see pls. 37, 38 and 39, where series of flowers from the same plants are illustrated). They are, however, much more alike than are the general run of individuals in a colony (see pl. 34). The most conspicuous differences are in regard to the petals. Those of figs. 9,5, and 6 (pl. 40) are unusually broad and flat, those 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 209 of number 7 are somewhat smaller, while number 8's are narrow and folded. There were accompanying differences in color which are only suggested by the photographs. Number 8, for instance, was distinctly lilac, while the others were blue. Similar results have been obtained in all the series of Iris versicolor which have blossomed to date. It is therefore concluded that Г. versicolor, though usually self-fertilized, is frequently cross-pollinated under natural conditions. The relation of this conclusion to the ques- tion of Jordanons or micro-species, is deferred to the Discussion. HYBRIDIZATION OF IRIS VERSICOLOR AND IRIS VIRGINICA A PRELIMINARY REPORT It has been found upon experiment that Iris versicolor and Iris virginica are partially fertile in crosses with each other. There has not yet been enough material in the breeding plot to make exact quantitative studies, but whereas nearly all the crosses which have been tried within either species have set seed, only Fig. 16. Ideographs of three hybrid plants; left, natural hybrid from Engadine, ch.; center, artificial hybrid produced at the Missouri Botanical Garden; right, natural hybrid from London, Ontario. slightly more than half of those between the two species have done so. The resulting hybrid seed has а low percentage of germina- tion and the seedlings show hybrid vigor. This was particularly noticeable when the young seedlings were grown in the cold-frame side by side with seedlings of the parent species. The seedlings [Уог. 15 300 ANNALS OF THE MISSOURI BOTANICAL GARDEN of Iris versicolor and of Iris virginica varied greatly in vigor, whereas the hybrid seedlings were all uniformly vigorous. Of the various crosses which have been made, two have flowered. They are reciprocal crosses between plants 1-ААА-3 and ТААС. I-AAA-3 is a plant of Iris virginica collected at Flint, Michigan, in 1923, and is illustrated in pl. 41, right figures. It is large- flowered and the flower has bright blue veining on a light back- ground. The petals are unusually broad. IAAG is a plant received from Dr. J. R. MeLeland of Pleasanton, Kansas, though it came originally from Medina, N. Y. It is a typical Iris versi- color and is outstanding in the dark color of its flowers and its stems. It is illustrated in pl. 41, left figures. The seedlings of cross IXB (I-AAA-3 x IAAG) were similar in size, leaf number, leaf height, etc., to the most vigorous plants of either parent species. The seedlings of the reciprocal cross, IXA ПЛАС XI-AAA-3) were even more vigorous. Plants one and two years old grown from seed showed leaves one fourth again as high and about half again as many leaves per plant as the parents. IXA and IXB flowered for the first time in 1927. Ideographs and photographs of their flowers and of those of the parent species are shown in fig. 16 and pl. 41. It should be emphasized that the measurements and pietures of IXA and IXB were made from plants growing in the cold-frame. 'They would have been much larger if grown in the water-side plots with the other plants. Thirteen plants of IXA and three plants of IXB were brought to flowering age. The two sets of hybrids were very similar aside from the greater vigor of IXA. While the seedlings varied slightly among themselves they were remarkably uniform in general aspect. They were intermediate in all the differences which characterize the two parents, though they were easily distinguished by their vegetative vigor. In general appearance and in practically all measurable characters they much more closely resembled Iris versicolor than Iris virginica. Their most outstanding differences from Iris versicolor were their much larger and laxer petals and the peculiar spots at the base of the вера]. The spots were due to the combination of the bright yellow pubescent spot of Iris virginica and the dark blue cross- 1928 ANDERSON—PROBLEM OF SPECIES IN IRIS 301 veining and brown stippling of Iris versicolor. The resulting “eye” is unlike anything in either species. Fig. 17. Cross-sections of sepals showing pubescence: A, I. virginica; B, I. X robusta; C, I. versicolor; Х 25. А microscopical examination of the pubescence of Ше sepal showed that of the hybrid to be intermediate in form between the two species though, due to the larger cells of the hybrid, it is almost as long as that of Iris virginica. ‘Camera-lucida drawings of all three are shown in fig. 17. The hybrid was likewise inter- mediate in the glandular development at the base of the stamens. Figure 18 shows cross-sections of the perienth tubes. об 18. Cross-sections of perianth tubes showing, a, style, and 6, glandular 2. of inner wall of tube; left, Iris versicolor, right, I. virginica, center, I. X robusta. ж The plants of crosses ЇХА and ЇХБ were partially sterile. Counts showed about 50 per cent abnormal pollen and many of the ovules aborted. Ovaries with one exposed locule are shown in pl. 41. Тһе plants from which the capsules were taken were growing in the same cold-frame and had had an equal opportunity for pollination. All of the seed capsules on the hybrid plants were small and shrunken. Numerous other hybrids between the two species have been [Vor. 15 802 ANNALS OF THE MISSOURI BOTANICAL GARDEN made, and while they have not flowered they are similar in their vegetative characteristics to those already described. Dr. J. R. McCleland, an iris amateur of Pleasanton, Kansas, has grown several crosses between the two species, some of which I have seen, and they are similar to [XB and IXA. Natural hybrids resembling those produced in the experimental plot have been found at several points where the ranges of the two species overlap. They would be commoner were it not for the barriers which exist between the two species; geographical barriers such as the extensive limestone areas west of the Alle- ghenies in which neither species is common; physiological barriers which prevent them from being wholly fertile with each other. The location of the hybrids and their relation to the distribution of the parent species is shown in fig. 19. Most of the hybrids found were very similar to those produced in the experimental plot. Those observed in Ontario resembled Ше garden-made hybrids very closely. There was little variation between them although they were found in several places. They were nowhere particularly abundant except in one creek bed just west of London, Ontario, where there was a large colony, appar- еп у all belonging to one clone. Iris virginica was fairly common throughout this district. ris versicolor was not found there though it was fairly common a short distance east of London. At Yale, Michigan, a single plant of Iris versicolor and a few hybrids were found. 'They were in а meadow near a small graveyard from which Г. versicolor may have spread. "There were no plants of Iris virginica in the immediate neighborhood, though it is common in eastern Michigan and there were large colonies within two miles. Ап extensive search of the neighborhood failed to locate any other plants of 1718 versicolor or of the hybrids. At Tawas City, Michigan, a single hybrid plant was found growing along the roadside among a group of plants of Iris versi- color. А colony of Iris virginica was found less than half a mile away. At only two points were the hybrids found making up large colonies. "These were both in the northern peninsula of Michigan, and though only a few miles apart they were so different in their composition that a separate description will be given of each. 1928] 308 PROBLEM OF SPECIES IN IRIS ANDERSON A + Т. versicolor.. ° a L. vieginica ©. I.versicolor v I.vieginica Fig. 19. Distribution of natural hybrids between J. versicolor and I. virginica (I. X robusta) in relation to that of the parent species. (Vou. 15 304 ANNALS OF THE MISSOURI BOTANICAL GARDEN WEST ST. IGNACE Along the main highway three miles west of Saint Ignace a very remarkable colony was located. It had apparently originated by the intermingling of the two species, followed by numerous crosses and back crosses. Тһе ideographs in fig. 20 give a slight idea of the bizarre mixture of types which was encountered; there was everything from typical versicolor to typical virginica with numer- ous intermediates and a number of very curious forms unlike any- thing seen before either in wild colonies or in the experimental plot. Most of the plants exhibited a high degree of vegetative vigor, though the environment did not seem to be unusually favorable. ENGADINE At Engadine, a few miles west of West St. Ignace another peculiar colony was found. It was most remarkable by reason of the great vegetative vigor of the plants. There was a degree of variation between the plants quite comparable to that found in colonies of the parent species, but by no means so extreme as that found in the colony at West St. Ignace. The plants were all very similar in their general aspect to the hybrids produced in the experimental plot. Ideographs of the colony show its lesser variability, and show the general similarity to the experimentally produced hybrid in fig. 16. Though admittedly difficult, if not impossible, to distinguish by means of ordinary herbarium material, the hybrid appears of sufficiently common occurrence to warrant a special name. X Iris robusta E. Anderson,! new hybrid (Iris versicolor X I. virginica) Intermediate between 1718 versicolor and I. virginica, but more robust; partially sterile. V. SUMMARY The common blue flags of eastern North America are found to be made up of two separate and distinct species; a northern and ‘ Iris robusta E. Anderson ris versicolor X I. virginica) Virginicam et versicolorem intermedia sed robustior, partialiter sterilis. ППППП ППАМА ППЛПП МАПП nnnnn nnnm nnnnn ААПАП A B Iris virginica Iris versicolor ШІ ШІ ШІ ШІ West St. Ignace, Mich. Engadine, Mich. Fig. 20. Ideographs of та. individuals each of the two by bey кем studied: C, West St. Ignace, Mich.; D, Engadine, Mich.; above, averag of I. virginica and 1. versicolor for comparison. [Vor. 15 306 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN eastern one, Iris versicolor L., and a southern and a middle- western опе, 1748 virginica L. Statistical studies have been made of 16 colonies of 1718 versi- color and 39 of Iris virginica. Each species is found to be a natural group. In spite of wide variation within each species there is no tendency whatsoever for one to merge into the other, and the general average of each species is practically the same wherever it is studied. Although variation between individuals is met with in every colony examined it has not resulted in any appreciable regional differentiation, since the species in question moved into their present home at the close of the glacial period. Iris versicolor and Iris virginica are kept separate in part by natural geographical barriers and in part by physiological differ- ences which prevent them from being perfectly fertile with each other. Along the narrow zone where the two species come in contact hybridization occasionally takes place. Natural hybrids, closely resembling those produced experimentally, have been found at five points. At two of these localities hybrid populations of considerable size had arisen and have been studied in greater detail. ‘There is some evidence that constant new forms might originate in this manner. VI. Discussion There are several objections which must be met before the above conclusions can be accepted as generally applicable. In the first place it might be argued that there were regional differ- ences present within the two species which were not revealed be- cause of the methods used, the characters chosen for study, etc. It is quite possible that such unrecognized differences existed, but if so they must have been of an entirely different order from the differences between species. The methods used did distinguish effectively between Iris versicolor and Iris virginica and did demonstrate that no such differences existed within either of the two species. For the material studied the conclusion therefore seems unavoidable that the differences within species are of an entirely different nature from the differences between them. A more valid objection to the general application of the con- clusions drawn in this work is that the two species belonged to an 1928] ANDERSON— PROBLEM OF SPECIES IN IRIS 307 old and well-defined genus and that different relations between species might be found to obtain in such a group as the genus Aster, for instance. From purely a priori reasoning it would seem quite possible that distinetions between species might be of а different nature in different parts of the vegetable kingdom. Conclusions drawn from a study of two species belonging to the Monocotyledons would certainly have to be confirmed with more closely related material before they could be accepted as valid for such a distantly related group as a genus of the Compositae. Another hindrance to the general application of the conclusions reached in the study of these two species 13 the geophysical nature of the region in which they occur. It is without great barriers of any sort. With a greater degree of isolation, such as would occur in à region cut by great mountain chains, the differences within species might perhaps be very different. It would be particu- larly interesting to compare the regional differentiation of the closely related western species, Iris missouriensis. LINNAEAN VS. JORDANIAN SPECIES 'The main conclusion drawn from this investigation is that the Linnaean species is a natural and permanent group. It should therefore be the most effective one for purposes of classification. In regard to Jordanian species I am in complete agreement with Clausen (727) when he says, “ The old botanists worked according to their own common sense and delicate biological feeling, but they had a conception of species far more nearly coinciding with what we now arrive at by careful statistical observation of var- iation in the field and by cytological investigations and crossing experiments than our modern small-species taxonomists. “Еше Art ist eine Art, ganz gleichgultig ob sie Diapensia lapponica, Viola tricolor, oder Hieracium margineiliceps heisst’, says du Rietz. I quite agree that it is a matter of supreme indifference to Nature what we decide shall be the definition of a species. But it is by no means a matter of indifference to ourselves whether we accept or reject a form of nomenclature which obscures Nature’s own chief system of division.” In my opinion those who have considered the Jordanon to be of prime importance, taxonomically and phylogenetically, have [Vor. 15 308 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN ascribed undue significance to the fact that it comes true from seed. With more genetical or horticultural experience they would have realized that coming true from seed (homozygosity) is a mere corollary of the amount of inbreeding which has taken place and that it is of minor taxonomic and phylogenetic significance. We can best demonstrate the actual significance of the Jord- anon’s true-breeding quality by examining the relationships between individuals in two different sorts of species; in one continuously cross-pollinated and in one continuously self-pol- linated. These relations are diagrammed in fig. 21 at a and b. Let us take four individuals of a continuously cross-polli- nated species, such a one as Aster anomalus, for instance, to name one that has actually been investigated (unpublished data). We may diagram these four individuals as A, B, C, and D (b, fig. 21) and if we examine them they will be found to possess certain inherent differences in such characteristics as the number, length, and shape of the ray flowers, the number and arrangement of the branches of the inflorescence, etc. Though two individuals may sometimes agree in a single characteristic, no two will possess the same combination of characters. In the next generation, however, none of these particular combinations will reappear. Plant A will be pollinated by pollen from other plants, as, for instance, by B as in the diagram, and the resulting offspring will show new combinations of characters unlike anything in the previous generation. In the diagram one of them is taken as an example and named L. Plants M, N, and O are similar new combinations arising from cross-pollinations between the other plants of the previous generation. There will be similar new recombinations of characteristics for each new generation. In other words, there will be just such a reshuffling of characteristics as occurs from generation to generation in human families; man being another species in which similar out-crossing prevails. If we now turn to a self-pollinated Linnaean species and study the differences between individuals we will find that they are very similar to those studied in Aster anomalus. We may therefore diagram them similarly (a, fig. 21) аз А, В, C, and D. In such a self-pollinated species, however, a single individual acts as father and mother for the next generation. There will ordinarily 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 309 be no crosses between A and В or В and С as there were in Aster anomalus. Furthermore, since such close inbreeding has obtained in the past, each individual will be practically homozygous (pure- breeding), and its progeny will resemble their parent and each other very closely. The seedlings of A will be so similar to their parent that they can be diagrammed as A, those of B will show the same combination of characters which dis- tinguished B from A, C, and D, and may be diagrammed as B This resemblance will continue from generation to generation as long as no cross-pollination occurs. Such a species is therefore divisible into a number of pure lines or Jordanons. | ; | a 6 b ig. Diagrammatic representation of the relationships between individuals in a, а self-pollinated species, b, а cross-pollinated species, and c, in J. versicolor It will be seen that the reappearance of distinctive combi- nations of characters from generation to generation is a mere corol- lary of the amount of inbreeding. Those Linnaean species which are continuously cross-pollinated will form new combinations in every generation. Those which are continuously self-pollinated will be divisible into recognizable pure lines or Jordanons. If species were of only these two sorts it might be possible to retain the Jordanon as an auxilliary unit, usefu. in the one case but not in the other. Unfortunately there is every transitional stage between the two extremes. 1748 versicolor and Iris virginica are species of this intermediate sort. Аз has been shown above, Iris versicolor is usually self-pollinated though occasionally out- crossed. The relationships between individuals may therefore be diagrammed somewhat as in c of fig. 21. Тће diagram may be taken as typical of many species which can be only partially [Vor. 15 310 ANNALS OF THE MISSOURI BOTANICAL GARDEN separated into Jordanons. Those individuals, such as plants A, B, C, and D in the diagram, which have resulted from a long line of self-pollinations will give rise to true-breeding strains as long as they are self-pollinated. But when a cross-pollination occurs, as when A is crossed with B, it produces new recombinations of characteristics. In the diagram one of these new individuals, L, is taken as an example. Even though self-pollinated it will not breed true and its offspring will vary among themselves. One of them is represented in the diagram and named T. Plant T will likewise be too heterozygous to breed perfectly true, and its progeny must be given a new symbol in the diagram. So even though continuous self-pollination occurs among the progeny of the cross between A and B, it will take six or more generations before pure breeding Jordanons will be established. Borrowing an idea from the diagrams in fig. 21, we may think of each species as a net, with the knots representing individuals. Those species in which cross-pollination regularly obtains will have a very short mesh, interweaving from one generation to the next. Those species, such as Iris versicolor, in which it is rare, will have a much longer mesh with fewer connecting cross-threads. Some few species will be so seldom cross-pollinated as to seem hardly net-like in their make up. They will be more like separate and unconnected knotted strings, but it is to be doubted if even in such extreme cases there never occurs a cross-pollination which brings the strings of the net together. Even though they occurred on the average only once in forty or fifty generations, it would cause the ultimate interweaving of the whole net and the disappearance of the single strings as separate units. The case for the Jordanon as a universally applicable unit becomes even more absurd when we consider the case of a species which is cross-pollinated in one part of its range and self-pollinated in another. While this has not been demonstrated for any wild species, there is no a priori reason why such a delicately balanced relation might not vary under different environments. Leighty and Taylor (727) have called attention to the fact that this has actually occurred in wheat which has different percentages of natural outcrossing in different parts of the world, as, for instance, in the Punjab where Howard and Howard ('09) report a very high 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 311 percentage. It has been found for numerous species that they are cross-pollinated only by a single insect. Such a Linnaean species, cross-pollinated in one part of its range and self-pollinated in another, would be divisible into Jordanons in the latter region but not in the former. The division of a Linnaean species into Jordanons is therefore seen to be a mere corollary of the amount of inbreeding which has obtained in that species. Where Jordanons do occur they owe their existence to continuous self-pollination and their individual- ity disappears as soon as a cross-pollination takes place. In those species where cross-pollination is of rare occurrence they will undoubtedly persist for some time, perhaps even for centuries, but they will ultimately disappear. These studies have shown that the Linnaean species, on the other hand, may retain its individuality, though submitted to widely differing environments, for long periods of time. 1718 versicolor and Iris virginica, for instance, must have persisted as recognizable units since they spread into their present homes at the close of the glacial period. Important as the Jordanon may be in the case of cultivated plants, or in those Linnaean species in which it naturally occurs, it is a relatively temporary unit of little taxonomic or phylogenetic significance. There remains to consider the bearing of the studies reported above on the question of the origin of species. For the material studied it has been shown that the differences between species are of an entirely different order from the differences between individuals. There is по evidence that these differences between individuals might, under the influence of natural selection or of any other natural force, eventually be compounded into differences compar- able to those between the two species studied. This conclusion is not in accord with most current speculation on the subject. Since the time of Darwin it has been very commonly supposed that the processes which give rise to differences between individuals, if allowed to operate over a longer period, will produce specific differentiation. This general theory has gone through many phases as biology has advanced; in its most recent form it is held by many of the Drosophila workers who see in the gene mutation the unit process which, compounded a thousand-fold, results in specific differences. Хот. 15 312 ANNALS OF THE MISSOURI BOTANICAL GARDEN There is little in the evidence reported i in this paper to support such an explanation of the origin of species. If all the individual differences which have occurred in 1748 virginica since the glacial period have not produced recognizable regional differences within the species, the production of new species by this means must be a very slow process. If we are to deny the slow accumulation of individual differences an important role in species building, to what process are we to turn? A number of recent investigators, Brieger (’28) Clausen (727), Karpechenko (727), to name only a few, have reported the experimental production of true-breeding hybrids as the result of new chromosomal realignments. The careful experimental inves- tigation of these hybrids has strengthened the case of those who believe with Lotsy (716) that hybridization has been an important factor in the evolution of species. Though it has not been investigated cytologically, the colony studied at Engadine is apparently composed of similar true-breeding hybrids of natural origin. While it will have to be very thoroughly investigated before it can be taken as conclusive, this apparent example of a new and constant form produced by the hybridization of two separate species is certainly very suggestive. VII. BIBLIOGRAPHY "een 25” (25). Monocotyledons. 250 pp. Cambridge, 1925. Babcock, E. B., and Hall, H. M. (24). Hemizonia congesta, a genetic, ecologic, 44 taxonomic study of the hay-field tarweeds. Univ. Calif. Publ. Bot. 13: 15-100. Bateson, У. (94). Materials for Ше study of variation, treated with especial regard to discontinuity in the origin of species. xvi+598 pp. London, 1894. Вперег, Е. (28). Ueber die Vermehrung der Chromosomenzahl bei dem Bastard Nicotiana tabacum Г. Х М. Rusbyi Britt. Zeitschr. f. Ind. Abstam.-u. Vererb. 47: 1-53. 1928. Butters, Е. К. ('27). Taxonomic studies in the genus Maianthemum. Minn. Studies in Pl. Sci. 6: 429-444. 1927. Clausen, J. (27). Chromosome number and the relationship of species in the genus Viola. Ann. Bot. 41: 677-714. 7» —————, (22). Studies on Ше collective species of Viola tricolor L. II. Bot. Tidsskr. 37: 363-416. 1922. dad L. (17). A consideration of the terms “вресїев” and “variety” as used ny, with special reference to the flora of New Zealand. N. Z. Inst., Trans. & Proc. 49: 66-79. 1917. ‚ and Allan, H. H. (27). The bearing of ecological studies in New Zealand on botanical taxonomic conceptions and procedure. Jour. Ecology 15: 234-277. 1927. 1928] ANDERSON—PROBLEM OF SPECIES IN IRIS 313 Hall, H. M., and Clements, Е. Е. (23). The e гр ај method in taxonomy. Сто Inst. Washington, Publ. 326: 346 pp. 1923. Harper, В. A. (28). The species concept from ibe point of view of a morphologist. Am. Jour. Bot. 10: 229-233. 3. Howard, A., and Howard, С. Г. С. (09). Wheat in India. 288 pp. Calcutta, 19 Jørgensen, С. А. (28). The experimental formation of heteroploid plants in the s Solanum. Jour. Genetics 19: 133-211. ol. 6-10. 1928. а а. D. (27). Polyploid hybrids of Raphanus sativus L. X Brassica oleracea Г. Bull. Appl. Bot., Genet. & Pl. Breed. 173: 305-410. 1927. Leighty, C. E., and Taylor, J. W. (27). Studies in ratural hybridization of wheat. Jour. Agr. Res. 35: 865-887. 1927. Lotsy, J. P. (16). Evolution by means of Ка ВИ 166 рр. The Hague, 1916. McLeland, J. В. (24). Beardless iris and how I became interested in them. Flower Grower 11: 91-93. McLeod, J. (26). The И method in biology. ххш+228 рр. Man- Needham, J. С. (00). Тһе fruiting of the blue flag (Iris versicolor L.). Am. Nat. Sumner, Е. В. (23). Some facts relevant to a discussion of the origin and inheri- tance of specific characters. bid. 57: 238-254. у Thompson, D. А. W. (17). On growth and form. 793 pp. Cambridge, 1917. Turrill, W. B. (25). Species. Jour. Bot. 63: 359-366. 1925 Vavilov, N. I. (27). Geographical regularities in the distribution of the genes of cultivated plants. Bull. Appl. Bot., Genet. & Pl. Breed. 17: 411-428. 1927. [Vor. 15, 1928 314 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 35 Fig. 1. Iris latifolia Virginiana, ек. Miller. From Ehret, С. D., Plantae Depic- tae, Tab. VI. 1748. Fig. 2. Iris latifolia Virginiana ete. Miller. From Ehret, С. D., Plantae De- pictae, Tab. VIII. 1748. PLATE 35 ANN. Мо. Вот. Garp., Vor. 15, 1928 " y», y y . 2227 ПМТ, сее e£ aeri GIO ‘DION YSTO ЕНТ = |4 L SINE preceumicns galu Жас. лата 5 пре байгсгг, баба in-s хайн бент. | / / | & "m е 722 РЕ, 18. "M fi 2 # ДРАВЕ, 4 eon a laters жаса ту ту Plot ул 72222 222 Stamina Mere gi ambiunt, ж n ја OW TER =. f 9% әт РТА tria 2577 21121277 | Арнан байн уйла ad. ети Fig 7222 72:12:31 Жк х < Fournefer таг Ж уйгу, ai “бишийн f ға Ж ситите PE % «tg V- Жон pir Éni stream «луге. / ж v Fructis hertcenta lier dfir. 2272 bá 7? 222 75799729 тет ее ті | а "ft 7, теще f алш. a ао) ме 2 mu | | 4%. Luna АУ | ! “ я ЗА тена; 72 ИНК ТИТУ ат 27, 72: a I Фра ‚жим ЊЕ жей 723 retin, we WAR "no 7727 езі Даррен 72 72112122 7, бау тут, аят 222 Ж айлийн 223 луу! Ver PR. M Каа nubis УНЕ | & © SEHH #1 ОА ИГРА ата, 1815 latifelua 2 ingenua Ha, «co vide ауу ГА ile tn tria folia peta rum асти 7 “езе a Germ л. 97227777 лубеит си „тен adhaeret „фита. trea. tl жұғуы malh Ж fr 77777772 га, unde Ager ab ennt : и. Ж "loy vy y ep ЕЕ Чета, 77777 "7 ^ х ғ 27 | f е ж (422 "Ито Ире ег ANDERSON—PROBLEM OF SPECIES IN IRIS Мог. 15, 1928] 316 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 36 I . Dillenius’ specimen of Iris americana versicolor, etc. лэг in the ОШешап Herbarium at Oxford. Fig. 2. [тїз carolina Radius. From Radius, W. M., Naturforsch. Ges. Leipzig Schrift. 1: 158. taf. III. БІНІ NI кпочав AO ИМТЧОЧА-ХОЯ8ЧЯаху ANN. Мо. Вот. Garp., Vor. 15, 1928 PLATE 36 «Т Ез ЗИН жуса Мог. 15, 1928] 318 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 37 Flowers of 1748 virginica, Х 14 Fig. 1. Plant ABC—collected at Orchard Farm, Мо. Fig. 2. Plant ABC—collected at Orchard Farm, Mo. Fig. 3. Plant ABC—collected at Orchard Farm, Mo. Fig. 4. Plant ABC—collected at Orchard Farm, Mo. Fig. 5. Plant ABC—collected at Orchard Farm, Mo. Fig. 6. Plant ABE—collected at Fort Madison, Ia. Fig. 7. Plant ABE—collected at Fort Madison, Ia. Fig. 8. Plant ABE—collected at Fort Madison, Та. Fig. 9. Plant ABQ—collected at Gilbertville, Ia. ANN. Mo. Вот. Garb., Vor. 15, 1928 Рълте 37 ANDERSON--PROBLEM OF SPECIES IN IRIS [Vor. 15, 1928] 320 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 38 Flowers of Iris versicolor, X М4 Fig. 1. Plant ABY—collected at Cohasset, Mass. Fig. 2. Plant ACF—collected at New Haven, Conn. Fig. 3. Plant ACF—collected at New Haven, Conn. 4, Plant BBG—collected at Harmonsburg, Pa. Fig. 5. Plant collected at Cedar Lake, Nova Scotia. 6. Partial albino, var. “Stella Main’’—collected in Connecticut. Figs. 7-9. Seedlings grown at the Missouri Botanical Garden from seed collected at Connecticut Lakes, N. H. ANN. Мо. Bor. Garb., VoL. 15, 1928 PLATE 38 ANDERSON—PROBLEM OF SPECIES IN [RIS 322 [У ог. 15, 1928] ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 39 Flowers of Iris virginica, X 14. “зай. 1-2. Plant ABB—collected at Valley Park, Мо. EA MAR Plant ACZ—collected at Fish Creek, Wis. Plant ABT—collected at Yale, Mich. Plant ABT—collected at Yale, Mich. Plant ABT—collected at Yale, Mich. Plant ABP—collected at Otisville, Mich. Plant ABP—collected at Otisville, Mich. Plant ABP—collected at Otisville, Mich. ANN. Мо. Вот. Garb., Vor. 15, 1928 PLATE 39 ANDERSON—PROBI EM OF SPECIES IN IRIS [Vor. 15, 1928] 824 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 40 One flower each of five sister plants of Iris versicolor grown at the Missouri Botani- cal Garden from a seed capsule collected at Connecticut Lakes, N. Н. ANN. Мо. Bor. Garp., Vor. 15, 1928 PLATE 40 ANDERSON—PROBLEM OF SPECIES IN IRIS [Vor. 15, 1928] 326 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 41 A comparison of Iris versicolor and Iris virginica with Iris X robusta, the hybrid between them Top row, immature capsules with one locule exposed. Middle row, immature capsules. Bottom row, flowers In each case the specimen at the left is from plant ТААС (Iris versicolor), that at the right from IAAA-3 (Iris virginica), and that in the center from their hybrid IXAM (Iris X robusta). ANN. Mo. Bor. Garp., Vor. 15, 1928 PLATE 41 ANDERSON—PROBLEM OF SPECIES IN IRIS [Vor. 15, 1928 328 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 42 Fig. 1. Typical flower stalk of Iris virginica. Fig. 2. Typical flower stalk of Iris versicolor. . dris virginica photographed at Huntingdon, Tenn. Fig. 4. Iris virginica photographed at Wilmington, N. C. ANN. Mo. Вот. Garb., Мот... 15, 1928 PLATE 42 ANDERSON—PROBLEM OF SPECIES IN IRIS 330 [Vor. 15, 1928 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 43 Figs. 1-5. Iris virginica. Fig. 1 Figs. Fig. Fig. Fig. Fig 6-10. Fig. Fig. Fig. Fig. Fig. . Collected seeds from nine localities, X 14. 2. Seed surface (St. Louis, Mo.), х 30. 3. Seed surface (Catawba, Ohio), X 30. 4. Seed, X 7. . 5. енд of seed capsule, Х 30. Iris versicolor. 6. Collected seeds from nine localities, Х №. 7. Seed surface (Sagamore, Мавв.), Х 8. Seed surface (Conewango, М. Y.), Х 30. 9. Seed, Х 10. Lining of тей сарвше, Х 30. ANN. Mo. Вот. Ganp., Vor. 15, 1928 1 PLATE 43 + > • «95b «ее >s д 4%» sre $06 941 во» eee ee 9 тез б К ФФЕе Фе» & ~ ә а 9 ада Albert Lea Мин. Denwisow, Mich. Franken oth, Mich. Port Moutow, N.Scotia Омен Sound, 0», Connecticut Lakes.NH 9%ө тээ ced » >» ата b^ aA у Фе Ф% Р ..% ьа 6 >» љ » 4.. «74 ээзач ье ө ъ ® Бе» а ээ Tawas City, Mich. Fostoria Mich. La Crosse, Wis. Steatford, М.Н. Or ford, МИ, Sagamore, Mast Ыз к» іа ГЕ > Фо ө 4096 във» » >ð + У e vooo я. ‚ е & bod »»э? Catawba, Ohio Anna, Illinois Bonwieville, Ky. Nantucket, Mass. Steuben Cr, NY Сопешапдо. N.Y ` ANDERSON—PROBLEM OF SPECIES IN IRIS [Уоь. 15, 1928] 332 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 44 Fig. Upper row, two plants of Iris virginica from Valley Park, Mo., grown in cinders ииги railroad track; lower row, two plants from the same colony but growing in rich swamp. Fig. 2 Large single clone of Iris virginica, dim N. C. Fig. 3. Colony of Iris virginica at Camden, Tenn 44 PLATE 15, 1928 ARD., VOL. ч 1 Амм. Мо. Вот. ( ANDERSON PROBLEM OF SPECIES IN IRIS Annals of the Missouri Botanical Garden Vol. 15 NOVEMBER, 1928 No. 4 A NEW VARIETY OF VERNONIA LINDHEIMERT: ESTHER L. LARSEN Instructor in Botany in the University of Montana Formerly Jessie R. Barr Fellow in Botany, Washington University A very interesting Vernonia was collected by Roxana S. Ferris and Carl D. Duncan along the Sanderson-Sheffield Road, twelve miles from Sanderson, Terrell County, Texas, July 19, 1921. The plant was distributed as Vernonia Lindheimeri Gray & Engelmann; but upon critical study and comparison with a relatively large suite of specimens representing this species in the Missouri Botanical Garden Herbarium, the Ferris and Dun- can plant shows such marked variation from the type of the species that it seems worthy of recognition as an outstanding variety. A description is recorded as follows: Vernonia Lindheimeri Gray & Engelmann var. leucophylla Larsen, n. var. РІ. 45. Formae typicae habitu simili; foliis utrinque dense albido- tomentosis; involucri squamis lineari-lanceolatis, acutis, dense tomentosis, marginibus purpurascentibus; achaeniis circiter 4 mm. longis, glabris. Suffruticose, stem densely tomentose; leaves linear to linear- lanceolate, 4-16 cm. long, 3-8 mm. broad, entire, densely to- mentose on both surfaces, margins of the younger leaves occa- sionally revolute; inflorescence terminal, branching, spreading, leafy; heads 12-17 mm. high; involucre broadly campanulate, 4-5-вегізде, 8-9 mm. high and 8-10 mm. in diameter; bracts linear-lanceolate, acute, purple-margined and densely tomentose; heads 50-60-flowered; flowers purple; pappus white; achenes 1 Issued December 22, 1928. Ann. Mo. Bor. Garp., Уог. 15, 1928 (333) [V ог. 15, 1928] 334 ANNALS OF THE MISSOURI BOTANICAL GARDEN about 4 mm. long, glabrous and glandless or rarely bearing a few glands in the furrows.—T'Exas: collected along the Sander- son-Sheffield Road, twelve miles from Sanderson, Terrell County, July 19, 1921, Кохапа S. Ferris & Carl D. Duncan 2826 (Мо. Bot. Gard. Herb. No. 902145 түре). Vernonia Lindheimeri Gray & Engelmann has been recorded heretofore only from Texas. It is noteworthy, however, that specimens of this species were collected in pine woods about Texarkana, Arkansas, August, 1881, by the late Mr. George W. Letterman. This collection extends considerably the geograph- ical range of the species, which, from material in the Missouri Botanical Garden Herbarium, may now be given as from south- ern Arkansas in the region of Texarkana southwestward to Sweetwater and San Antonio, Texas. The variety leucophylla at the present time is known only from Terrell County, Texas, which is about 200 miles west of the westernmost station recorded for the species. The distri- bution areas of Vernonia Lindheimeri and its variety leucophylla in all probability on further exploration will be found to overlap. EXPLANATION OF PLATE PLATE 45 Vernonia Lindheimeri Gray & Engelmann var. leucophylla Larsen Texas From the type specimen, Ferris & Duncan No. 2826, in the Missouri Botanical Garden Herbarium. ANN. Mo. Вот. Garp., Vor. 15, 1928 PLATE 45 С +3 ; Ч Lar у xoc ҮХЭЖ А Roxana 8. Fefris and Сан D. Duncan July If, на аж», лс phat 1 b i Lanes ў darer ЗОР Чы: LARSEN—NEW VARIETY OF VERNONIA LINDHEIMERI DYSOSMA: A NEW GENUS OF BERBERIDACEAE! ROBERT Е. WOODSON, JR. Rufus J. Lackland Research Fellow in the Н 52 Shaw School ој Botany of Washington Universit While examining herbarium material 8 Ше genus Podophyllum in the Gray Herbarium of Harvard University recently, the writer happened upon six sheets of a very curious plant col- lected in China by Henry in 1854, and also two sheets of similar specimens collected by Ford, and distributed from the Hong- kong Botanic Garden in 1885. The plants were immediately perceived to be radically dis- tinct from both the North American P. peltatum L. and the Asiatic P. Emodi Wall. Тһе evident differences are larger size in general, much broader leaves with very shallow and regular lobing, and especially an umbel of four to nineteen flowers instead of the familiar solitary flower of the more common species. Upon a consultation of the literature, it was found that the plants correspond to the description of P. versipelle Hance. A somewhat closer superficial examination disclosed the facts that the pedicels of the inflorescence are distinctly recurved, while those of the more familiar species are erect or slightly nodding, and that the petals are also drooping, oblong-lanceolate in outline, and of a dull reddish color. A difference in the rhizome was also evident, it being thick and fleshy and desti- tute of scales, with the nodes crowded together almost as a tuber, while the rhizomes of both P. peltatum and P. Emodi are more like the ordinary creeping stem, if such a distinction can be made, being slender, more fibrous, and giving rise to а con- spicuous production of cataphyllary scales (pl. 46, figs. 5 and 10). Dissections of the species in question disclosed further facts of considerable interest. The stamens of P. versipelle are four to six in number, fewer than in the other species, and are about one-half again as large in every dimension as those of both P peltatum and P. Emodi. The filaments are sharply curved 1 Issued December 22, 1928. ANN. Mo. Bor. Garb., Vor. 15, 1928 (335) ГУ он, 15 336 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN away from the pistil, unguiculate in a certain manner, and the sterile connective is greatly developed, apiculately produced at the apex, and bears the thin locules conspicuously extended in a parallel position from its ventral side. The anthers, more- over, are introrse, one of the most striking features of the species, since the anthers of all other Berberidaceae are extrorse, in- cluding those of P. peltatum and P. Emodi which are also pro- duced laterally from a narrow connective upon straight filaments. Upon an examination with a compound microscope the pollen of P. versipelle was found to be perfectly spherical, and about one-half to two-thirds the size of the lobed pollen of Р. peltatum and P. Emodi. The pistil of P. versipelle also produces a definite slender style bearing a globose stigma, while the other two species have peltate stigmas which are sessile or only slightly elevated. The mature fruit of P. versipelle is unknown, but it is presumed to be rather similar to the pulpy bacca of P. peltatum and P. Emodi. With two such distinct elements as are represented by P. versipelle, on the one hand, and P. Emodi and P. peltatum, on the other, it appears that the equilibrium of the Berberidaceae and the tribe Podophylleae, containing at present only the genus Podophyllum, should be more easily maintained by recognizing the elements as distinct genera of a single tribe, since two such genera would be quite as distinct in their separate tribe as those of the other tribes of the family, as, for instance, Berberis and Mahonia, and Epimedium and Vancouveria in the Berberideae. In establishing the new genus, the name Dysosma has been constructed from the Greek 80$ + ёошў, signifying “а disagree- able odor," chosen arbitrarily upon the testimony of Hance, who was able to examine fresh plants, and who pronounced their odor as most remarkably putrid. Tabulated, the differences of Dysosma and Podophyllum have been found to be as follows: DYSOSMA PODOPHYLLUM шы tuberous, without cata- Rhizome a creeping slender stem, phyllary scales or prophylls. with both cataphyllary scales and prophylls. Flowers in umbels. Flowers solitary. 1928] WOODSON—DYSOSMA: NEW GENUS ОЕ BERBERIDACEAE 337 DYSOSMA Pedicels reflexed. Petals drooping, dull reddish. Pistil with a definite style. Stigma globose. Stamens 4—6, introrse. Stamens with an enlarged sterile connective. Filaments unguiculate, spreading away from the pisti Leaf-lobes shallow and regular, sharply and regularly denticu- ate. Pollen spherical, relatively small. PODOPHYLLUM Pedicels erect or only slightly nodding. Petals spreading, white. Pistil without a definite style. Stigma peltate. Stamens 6-18, extrorse. Stamens without an enlarged ster- ile connective. Filaments straight, not spreading. Leaf-lobes deep and irregular, en- tire or irregularly laciniate. Pollen lobed, relatively large. An examination of the literature of the many-flowered Podo- phyllums published from Asia discloses the fact that six species have been described, differing from one another by dissimilarities strikingly analogous to those variable characteristics frequently found in P. peltatum and P. Emodi, such as the position of the flowers, pubescence, lengths of stamens, and even carpellary number. The specific features of P. Veitchii and Р. difforme, for example, are stamens slightly longer than the petals, and stamens half as long as the petals, respectively; supposed to differ from P. versipelle, which is presumed to have stamens and petals of equal length. Upon examination of herbarium? material the writer has been fortunate to find a specimen with two flowers remaining upon the pedicels of the inflorescence, one with stamens longer, and the other with stamens somewhat shorter than the petals (Henry 5872F, MBG). Likewise, petals have been found to be notched in a manner similar to that described for P. Onzoi. Р. Esquirolii, furthermore, is said to 1 А paper dealing with the morphological variability and the involved synonymy which it has produced in the genus Podophyllum is now in manuscript. 2 In the taxonomic treatment which follows, the herbaria from which exsiccatae have been cited are abbreviated as follows: Gray Herbarium (GH); Missouri Bo- tanical Garden Herbarium (MBG); New York Botanical Garden Herbarium (NY); United States National Herbarium (US). The writer desires to express his appre- ciation for the facilities which were kindly allowed him by the respective curators ach. (Vou. 15 338 ANNALS OF THE MISSOURI BOTANICAL GARDEN have leaves which are almost without lobing, a feature which might be explained by the great leaf variability of P. peltatum and Р. Emodi. Although predominately extra-axillary, the ш- florescence of Dysosma has been occasionally found to be axil- lary. Although perhaps taking too much liberty in doing so, it has been thought advisable in the establishment of the new genus to treat the species described upon such characters as have been found spontaneously variable in P. peltatum and P. Emodi as representing variations of a single species. Since axillary forms have been reported only from Formosa, it may well be that the position of the umbel is not variable and that the axillary forms are specific. Until better knowledge is avail- able, however, the extra-axillary form is taken as the normal, and the axillary as the abnormal form. Although future study шау hold other species genuine, only one, the oldest, which happens to be P. pleianthum Hance, has been retained and transferred to Dysosma. Dysosma! n. gen. Herbaceous caulescent perennial, glabrous or somewhat pu- bescent. Rhizome indeterminate, thickened, fleshy, without cataphyllary scales or prophylls. Leaves 1 or 2, peltate, pal- mately lobed, the lobes regular, usually 6 large anterior and 2 smaller posterior lobes, regularly denticulate. Flowers in axil- lary or extra-axillary umbels of 4-19, the pedicels reflexed. Petals usually 6, oblong-lanceolate, dull reddish, drooping. Se- pals 3, petaloid, fugaceous. Stamens usually 6; filaments long, unguiculate, spreading from the pistil; anthers 2-celled, dehiscing longitudinally, ventrally parallel, produced from an enlarged ‘Dysosma Woodson gen. nov. Berberidacearum, herba perennis ми erecto glaberrimo pruinoso 3—5 dm. alto; foliis radicalibus solitariis caulinis binis crassius- culis centrice vel subcentrice peltatis orbiculatis palmatim 6-8-lobatis, lobis late triangulo-oblongis acuminatis vix quintam diametri partem aequantibus, margine creberrime subulato-denticulatis, petiolis pruinosis aequilongis; pedicellis declinatis; floribus 4-19 ad apicem caulis infra foliam superius petiola vel inter folia apice eaulis nascentibus, ebracteatis; sepalis 3 tantam deciduis; petalis 6-9 oblongis acutis sordide sanguineo-rubris 1.5 em. longis; stamine 4-6 antheris valvula longi- tudinali utrinque dehiscentibus introrsis, filamentis unguiculatis aequilongis con- nectivo ultra loculos in apiculum producto; ovario ellipsoideo-sphaerico gracilibus stigmateque globosis cristatis coronatis, ovulis indefinitis. 1928] WOODSON—DYSOSMA: NEW GENUS ОЕ BERBERIDACEAE 889 apiculate sterile connective, introrse. Ovary oblong in outline, l-celled; ovules many, anatropous, each enclosed in a fleshy aril, disposed upon a lateral placenta; stigma globose, thick, produced upon a definite style. Mature fruit unknown, prob- ably a fleshy berry. Type, Henry 5372F, Sze-chuan, China, 1885-88 (MBG). Type species: Dysosma pleiantha (Hance) Woodson. 1. Dysosma pleiantha (Hance) Woodson, n. comb. Pl. 46. Podophyllum pleianthum Hance, Jour. Bot. 21: 175. 1883. Podophyllum versipelle Hance, l. с. 362. 1883. Podophyllum Veitchii Hemsl. & E. Н. Wils., Kew Bull. Misc. Inf. 1906: 152. 1906. Podophyllum а отте Hemsl. & E. H. Wils., l. с. 1906. Podophyllum Ездштош Léveillé in Fedde, Керегі. 11: 298. 1912. Podophyllum Onzoi Hayata, Icon. Pl. Form. 5: 2. 1915. Characters of the genus. Distribution: southeastern China and the island of Formosa. Specimens examined: СнтмА: Hupeh, April, 1885, Henry 8952 (GH, US); data lack- ing, Hongkong Botanic Garden, Ford (GH); Canton, Lo-fanshan Mts., date lacking, Ford 1092 (US); Sze-chuan, 1885-88, Henry 5872 (СН, MBG түре, NY, US); Chekiang, 1908, Barchet 22 (US); Chekiang, 1906, Barchet (US); western Hupeh, May, 1907, Wilson 3202 (US). [Vor. 15, 1928] 340 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 46 Comparative morphology of Podophyllum and Dysosma. Fig. Habit of Podophyllum Emodi Wall. Fig Receptacle of P. Emodi with pistil and four stamens. Fig Stamen of P. Emodi. Fig Diagrammatic cross-section of stamen of P. Emodi. Fig Rhizome of P. Emodi. Habit of Dysosma pleiantha (Hance) Woodson. Receptacle of D. юэ" with pistil and two stamens. Stamen of D. pleiant Diagrammatic 5, of stamen of D. pleiantha. Rhizome of D. pleiantha. = у я 03 08030 Өюөноөсоюоюн = ^ — Ann. Мо. Вот. Ganp., Vor. 15, 1928 PLATE 46 WOODSON—DYSOSMA STUDIES IN THE APOCYNACEAE. П! A REVISION OF THE GENUS STEMMADENIA ROBERT E. WOODSON, JR. Rufus J. Lackland Research Fellow in the Henry Shaw School of Botany of Washington University HistoricaL REVIEW The Apocynaceous genus Stemmadenia was established in 1844 by Bentham,? who recognized three species, the type, 5. glabra, 8. pubescens, and 8. mollis. The first and third spe- cies were entirely new to science, but the second Bentham per- ceived to be identical with Bignonia ? obovata Hook. & Arn., although he chose to give the species an original name. The specific adjective of Hooker and Arnott has subsequently been restored by Schumann.‘ In 1853, A. Richard,* evidently unaware of Bentham’s genus, published the genus Odontostigma, with one species, O. Galeot- папа, from the environs of Havana, Cuba. From the evidence of an excellent plate which illustrates Richard's genus, Miers,* in 1878, was able to definitely identify Odontostigma as герге- senting merely another element of Stemmadenia. Thirty-four years after the establishment of the genus by Bentham, Miers’ presented a treatment of Stemmadenia in his monograph of the South American Apocynaceae. In the treat- ment of Miers, besides the three species of Bentham, five new 1 Studies in the Apocynaceae. I, containing an historical account of the taxonomy of the family and a critical study ‘of the tribe Apocyneae, is in manuscript, and will appear in a subsequent number of the ANNALS OF THE MissouRI BOTANICAL GARDEN. з Benth. Bot. Voy. Sulph. 124. t. 44. 1844. 3 Hook. & Arn. Bot. Beechey’s Voy. 439. 1841. Concerning the mistake of the Е for а Bignonia, Bentham wrote: “А portion of Ше seed vessel and a Pithecoctenium, probably P. muricatum, had been by mistake laid by Dr. Sinclair into the same sheet with the specimen of this plant, and had misled the authors of the ‘Botany of Captain Beechey’s Voyage’ and induced them to refer the plant E to Bignonia." Benth., !. c. 125. 1844 5 п Engl. & Prantl, Nat. Pflanzenfam 42: 149. 1895. PA. Rich. | in Sagra, Hist. Cub. 11: 868. t. 5 1853. 5 Miers, Apoc. S. Am. 76. 1878 71, с. 74-77. À Issued December 22, 1928. ANN. Мо. Bor. Garb., VoL. 15, 1928 (341) Vor. 15 342 ANNALS OF THE MISSOURI BOTANICAL GARDEN species are added to the genus, namely, S. grandiflora (Tabernae- montana grandiflora Jacq.), S. insignis, S. Galeottiana (Odonto- stigma Galeottiana A. Rich.), S. bella, and S. bignoniaeflora (Echites bignoniaeflora Schl.). Of these, the most important by all means is S. grandiflora, which introduced a very distinct element into the genus and which, in this revision, is considered to merit subgeneric distinction. The work of Miers, which was the last to review the genus, was largely but a compilation of the descriptions of plants which the author himself had never seen, and since he had been able to examine only three of the eight species which he recognized, his product is liable to frequent errors, to obviate which will be in part the duty of this revision. Since the treatment of Miers, several species have been added to the genus, and explorative activity in Central and South America has greatly augmented representatives of the genus in herbaria. In the course of recent determinative work on miscel- laneous American Apocynaceae, an encounter with the technical and nomenclatoral difficulties of Stemmadenia has convinced the author that a revision of the genus might appropriately be introduced into this series of Studies in the Apocynaceae. The study entailed in the preparation of this revision was begun at the Gray Herbarium of Harvard University and com- pleted at the herbarium of the Missouri Botanical Garden. The author desires to express his appreciation to Dr. B. L. Robinson and to Dr. J. M. Greenman for assistance and sug- gestions during the course of the study, and to Dr. George T. Moore for the privileges of the Missouri Botanical Garden. He is also indebted to Dr. N. L. Britton and Mr. Percy Wilson, of the New York Botanical Garden, Dr. Е. W. Pennell, of the Philadelphia Academy of Natural Sciences, Dr. W. R. Maxon and Mr. Е. Р. Killip, of the United States National Herbarium, and to Mr. P. C. Standley and Mr. J. F. Macbride, of the Field Museum, for the courtesy of study in the various herbaria. GENERAL MORPHOLOGY The various species of Stemmadenia are shrubs or small trees attaining a height of two to twelve meters. 1928] WOODSON—STUDIES IN APOCYNACEAE. II 343 Leaves.—The leaves of the genus are opposite, membrana- ceous, entire, penninerved, glabrous or pubescent, and petiolate. The sheaths of the petioles are conspicuous, meeting in a shallow ring about the stem. Numerous fusiform glands are concealed in the petiolar ring of the leaves, but are fully exposed and persistent when the leaf drops from the stem. Тһе presence of these glands has been overlooked by each previous student of the genus. The outline >f the leaf varies little, the most frequent form being ovate-oblong. However, variations occur in the spathu- late leaves of S. Donnell-Smithiz and the lanceolate leaves of S. eubracteata. The surface of the leaves is extremely variable, and may grade upon the same specimen of certain species from tomentose, through barbate, to glabrous. In the case of other species, however, the surface of the leaves is relatively constant. The length of the petiole appears of some constancy, and is occasionally used as an accompanying taxonomic criterion. Inflorescence.—The inflorescence is a reduced terminal суше, bearing from one to several flowers usually, and three incon- spicuous bracts upon the pedicel of each flower. The ordinary well-developed inflorescence of the majority of species of the genus produces four to ten flowers, but an exception is found in the case of S. pauciflora which normally develop but one flower for each cyme, although one to several abortive buds may appear. The bracts usually directly subtend the flower, but in certain species, as in S. eubracteata, may appear about midway upon the pedicel. The character of the bract is a differentiating cri- terion between the genera Tabernaemontana and Stemmadenia, since in the former genus the bract always subtends the pedicel or aborts entirely. alyx.—The calyx consists of five imbricate lobes of unequal size, the three interior being somewhat larger, and usually more nearly colorless than the two smaller exterior lobes. Upon the interior of the calyx-tube, near the attachment to the disc, are borne several cycles of small fusiform glands, which may vary in approximate number from fifty to over one hundred. The unequal lobes of the calyx and the unusual number of the caly- cine glands are obvious distinguishing marks of the genus. [Ү о. 15 344 ANNALS OF THE MISSOURI BOTANICAL GARDEN The relative length of the calyx-lobes and the general size of the calyx are of basic importance in the speciation of the genus, and form at once an evident, and it is believed a reliable and natural, taxonomic criterion. The lobes may vary from 2 cm. in some species to 1 mm. long in others, and are usually distinct for the various species. Corolla.—The genus Stemmadenia is at once divisible into two subgenera largely upon the basis of the form of the corolla. The sections are also based upon this character. The corolla is salverform in the subgenus Ochrodaphne, and infundibuliform in the subgenus Hustemmadenia. The salverform corollas of Ochrodaphne are fairly regular, but the infundibuliform corollas of Ки еттадета divide into two series, namely, that of the section obovatae, with a conical proper-throat and a spirally twisted tube, and that of the section Galeottiae, with a cylindrical proper-throat and a tube without spiral twisting. The relation of the proper-throat to the proper-tube of the infundibuliform corollas is again apparently a matter of taxonomic importance in the case of certain species, as is also the length of the limb. Within the corolla-tube, above and opposite the attachment of the stamens, are five conspicuous appendiculate folds which vary considerably in length, but are constant for the genus. The corollas are large and showy, and are either yellow or yellowish white in color. The five equal lobes of the limb are dextrorsely deflexed, especially in the subgenus Ochrodaphne. Stamens.—The five stamens are wholly inserted, and are attached to the corolla-tube by short, thick, unguiculate fila- ments. The two sporangia comprising the anthers are elongate- fusiform in shape, and may be practically parallel, as in the subgenus Ochrodaphne, or obviously divergent at the base of the anther, as in the subgenus Еизеттадета. Тһе anthers are entirely fertile and unappendaged. Pistil.—The pistil is typically bi-carpellate. The carpels аге sessile and are separate except at the apices, which connive to form the filamentous style. Each carpel is uniloculate and contains many ovules upon a binate ventral placenta. The stigma is borne upon a fleshy terminal clavuncle. Disc.—The dise proper is inconspicuous, shallow, and im- 1928] WOODSON—STUDIES IN APOCYNACEAE. 1 345 mersed, but is surmounted by a ring of five conspicuous fleshy nectaries about the pistil, which, however, are actually coa- lesced into a more or less unified ring. The nectaries are par- tially adnate to the walls of the carpels, at least at the base. The nectaries appear of little taxonomic use. Fruit.—The fruit consists of a pair of divaricate, leathery, glandular-punctate follicles containing many striate, albuminous, ecomose seeds immersed ш an oily arilar pulp. The leathery pericarp eventually becomes coriaceous, and appears at that time to undergo a ventral dehiscence. The embryo is straight. It appears probable that were fruiting specimens of each species abundant peculiar diagnostic characters would be avail- able based upon the general shape and size, form of glandu- losity, etc. At present, however, the fruit of relatively few species is known, and in the following keys, the fruit is entirely omitted. SYSTEMATIC POSITION Concerning the affinities of the genus Stemmadenia, Richard was much better orientated than Bentham. Bentham,' in de- scribing the genus, wrote in part: “Тһе size and form of the flowers in the above three species |9. glabra, 8. pubescens, and S. mollis] are those of a Cerbera or a Thevetia from both of which, however, they differ in the calycine glands, and from the latter in the ovary; and in many points also there is a considerable degree of affinity with Odontadenia, but that genus again has not the remarkable calyx and glands of Stemmadenia А Since superficially all large flowers resemble one another, йй. tham was right in associating his new genus with Cerbera and Thevetia, although he does not mention the significant differ- ences between those genera and Stemmadenia. However, in referring to an affinity with Odontadenia the fallibility of the obvious is well demonstrated, for Stemmadenia, with unappen- daged anthers, non-connivent stamens, fleshy follicles. and eco- mose seeds, is about as distantly related to Odontadenia, with appendaged anthers, connivent stamens, chartaceous follicles, and heavily comose seeds, as two genera in the same family 1 Benth., 1. с. 125. 1844. Vor. 15 346 ANNALS OF THE MISSOURI BOTANICAL GARDEN could be. More recently Miers, evidently deceived by the external similarity of the flowers of Stemmadenia to the showy flowers of the Echitoideae, pictured the stamens of S. insignis with conspicuous basal appendages. Richard? displayed an understanding view of the morphology of Apocynaceous genera when, in describing Odontostigma, he remarked ‘‘Difiere del genero Thevetia sobre todo por su caliz, mas ancho y mas largo y por sus ovarios distintos, conteniendo cada uno gran numero de ovulos y no dos ovulos solamente como en el genero Thevetia.” Miers, in following Richard’s carpological view of the subject, has justly associated Stemma- denia with Tabernaemontana, its nearest relative, but has evi- dently failed to make sufficiently clear the differences which exist between them. In summing up the results of recent study, it is clear that Stemmadenia, by reason of its unappendaged anthers and non- connivent stamens, is a member of the subfamily Plumeroideae of Apocynaceae. Furthermore, by reason of its two carpels forming a divaricate fruit, it belongs to the tribe Plumereae. Finally, the fleshy follicles ally the genus immediately with the genera Cerbera, Thevetia, Vallesia, and Tabernaemontana in the subtribe Tabernaemontaninae. From the genera Cerbera, Thevetia, and Vallesia, Stemmadenia differs, as Bentham and Richard have indicated, in the nature of the calycine glands, which are so conspicuously multiplied іп Slemmadenia, in the calyx, which is conspicuously irregular іп the latter genus and regular in Cerbera and Thevetia and Vallesia, and in the fruit, which is monospermous in the latter three genera and polyspermous in Stemmadenia. Finally, it is noteworthy that the carpels in Cerbera and Thevetia develop together, while those of Stemmadenia become widely divaricate, in which character it appears related to Vallesia. From the genus Vallesia, Stemmadenia also differs in the corolla, which is much larger than in the former genus, and in the inflorescence, which is more reduced. The fleshy pericarp of Vallesia, also, is watery and evanescent, differing from the leathery persistent pericarp of Stemmadenia. 1 Miers, l. с. pl. 10B. 1878. 2 Richard, 1. с. 1853. 1928 WOODSON-—STUDIES IN APOCYNACEAE. II 347 The differences between T'abernaemontana and Stemmadenia have not always been easy to perceive. The most conspicuous difference is in the size of the flowers, which is much greater in the latter genus than in the former. However, technical char- acters are several and concise. The irregularity of the calyx of Stemmadenia again sets it apart from the regular calyx of Табет- naemontana. The interior of the corolla-tube in Табегпаетот- tana is naked, and contains appendiculate folds in Stemmadenia. The calycine glands of the former are uniseriate, while those of the latter are multiseriate. The nectaries of the former are completely coalesced and adnate to the carpels; the nectaries of the latter are only partially coalesced and are scarcely adnate to the carpels. The filaments of the former are straight, while those of the latter are unguiculate. Also it is believed that the fruit of Stemmadenia, which is much larger than that of Tabernaemontana, is eventually dehiscent along a ventral su- ture, while that of the latter genus is always indehiscent. The corolla of Stemmadenia is infundibuliform, or, if salverform, the tube is spirally twisted and the calyx is immediately subtended by three bracts, while in T'abernaemontana the corolla is always salverform, although the tube is not spirally twisted and bracts are limited to one or two, which subtend the pedicel rather than the calyx itself or are lacking. RELATIONSHIPS WITHIN THE GENUS As has already been explained, the genus Stemmadenia is readily divisible into two subgenera. The subgenus Hustemma- denia comprises plants with infundibuliform corollas, the lobes of which are slightly deflexed dextrorsely. The calyx squamellae are in several series. The sporangia of the anthers are diver- gent at the base. The species of the subgenus Ochrodaphne possess salverform corollas with lobes which are conspicuously deflexed dextrorsely, and auriculate, giving the flower a striking turbinate appear- ance. The calyx squamellae are fewer than those of Hustemma- denia and are usually in only two or three series. The sporangia of the anthers are nearly parallel to the base. The pistil of Ochrodaphne, also, represents a condition much [Vor. 15 348 ANNALS OF THE MISSOURI BOTANICAL GARDEN nearer coalescence of the carpels than that of Еизеттадета. The carpels of Eustemmadenia are prolonged into two distinct stylopodium-like beaks before they finally unite into а common style bearing the stigmatic clavuncle. On the other hand, the 2А е Fig. 1. СагреПагу diagrams of Stemmadenia. 1а, pistil of 8. tomentosa var. Palmeri; 1b, pistil of S. grandiflora; 1c, cross-section of ovary of S. tomentosa var. Palmeri; 1d, cross-section of ovary of S. Galeottiana; Те, cross-section of ovary of S. grandiflora. style of Ochrodaphne arises directly from the summit of the truncate carpels. Diagrams of the pistils of Hustemmadenia and Ochrodaphne are found in fig. 1, which also diagrams a differ- ence in placentation occurring between the two subgenera. Eustemmadenia appears to be the more primitive of the sub- genera and Ochrodaphne the more advanced because of the form of the corollas and pistils of those groups, and also by reason of the reduced calyx-squamellae of the latter. In a future paper reasons for assuming the floral squamellae frequently occurring in the Apocynaceae as staminal vestiges will be fully discussed, and until then the reasons for regarding the reduc- tion of squamellae as a modified rather than a primitive state must remain implied. For morphological reasons which have already been advanced, Stemmadenia is apparently more primitive than its closest neigh- boring genus, Табегпаетотата, and should therefore logically be placed after that genus in a phylogenetic synopsis of the family Apocynaceae. At present, in the system of K. Schu- 1928] WOODSON—STUDIES IN APOCYNACEAE. II 349 mann in Engler and Prantl’s ‘Naturlichen Pflanzenfamilien,’ this order is reversed. This latter order is also found in the ‘Genera Phanerogamarum’ of Dalla Torre and Harms. ‘The logic of viewing Stemmadenia as more primitive rather than more advanced than Tabernaemontana is perceived when the genus is split into the two subgenera, Hustemmadenia and Ochro- daphne, indicating an advance from a simple corolla and numerous squamellae to a more highly modified corolla and reduced squamellae, including an advance in the coalescence of the carpels, tendencies finally developing a climax in the mor- phology of the genus T'abernaemontana. The species within the group Ochrodaphne are homogeneous and not divisible into subgroups, but the species of Еизетта- denia are clearly divisible into two sections, illustrating an approach from the infundibuliform corolla characteristic of the subgenus to the salverform corolla of Ochrodaphne. Of these, the obovatae possess a typically infundibuliform corolla with a nearly conical proper-throat and a dextrorsely twisted tube, while the Galeottiae possess a modified form of infundibuliform corolla with a long cylindrical proper-throat, and a tube which is without dextrorse spiral twisting. Apparently the only other morphological difference which accompanies the corollar char- acters of these sections is the amount of space left between the carpels as an index of the degree to which carpellar fusion has progressed. Figs. 1с and 1d illustrate diagrammatically cross- sections of the pistils of S. tomentosa var. Palmeri and S. Gale- ottiana, representatives respectively of the sections obovatae and Galeoitiae. It is easily perceived that the carpels of the latter are the more nearly coalesced, which corroborates the judg- ment of it as the more advanced, phylogenetically. The carpels of the subgenus Ochrodaphne, as fig. le testifies, are at about the same stage of coalescence as those of the Galeottiae section of Eustemmadenia. GEOGRAPHICAL DISTRIBUTION The genus Stemmadenia is confined apparently to the tropical regions of continental America, lying between the Equator and the Tropic of Cancer roughly, although it also occurs slightly more to the north of those arbitrary bounds as far as southern (Vor, 15 350 ANNALS OF THE MISSOURI BOTANICAL GARDEN Chihuahua in Mexico, and doubtless also farther to the south, especially in Ecuador. The species of the genus are frequenters of sub-Cordilleran underbrush, and range in height from two to twelve meters for mature specimens. ‘The fruit is said by Miers to constitute a favorite food for the larger birds of the region, and seeds are probably distributed by means of those agents. As fig. 2 indicates, the subgenus Еизеттадета and the sub- genus Ochrodaphne coincide in their ranges in Central America, but have distinctive ranges, Hustemmadenia towards the North, and Ochrodaphne towards the South. However, S. obovata var. mollis, one of the most widespread and common representatives of Hustemmadenia sect. obovatae, has twice been collected about Guayaquil, Ecuador, and once near Yungas, Bolivia, imparting a most singular appearance to a map of the distribution of the genus. The disrupted nature of the distribution of the sub- genus Husltemmadenia thus disclosed urges a consideration of it as a relict group; thus as in all probability the more primitive of the subgenera, and the section obovatae as the ancestor of the entire group, even as a study of the morphology alone indi- cated. Although Richard, in describing Odontostigma Galeottiana, stated that the specimens were from the environs of Havana, Cuba, and although one would naturally expect to find repre- sentatives of a genus so frequent naturally in Central America in the Antilles, no evidence of the presence of the genus in Cuba or the other Caribbean Islands has been found, either in herbaria or in published floras of the region. It appears probable that Galeotti’s specimen from which Richard drew his description was in reality collected in Mexico, rather than in Cuba, as was understood by Richard. ABBREVIATIONS In citing specimens, the following abbreviations for herbaria have been employed: G = Gray Herbarium of Harvard Uni- versity; NY = Herbarium of the New York Botanical Garden; US = United States National Herbarium; ANSP = Academy of Natural Sciences of Philadelphia; F = Herbarium of the Field 1928} 351 STUDIES IN APOCYNACEAE. II WOODSON озиэрршлшэ)с! 10 seeds jo поцпатдвтр Зшмочв- 72 `2] у ! 20 х G 2. ! t не ae 1 |: “+ (954 4 М” а; ~ о ~ ~ e PEOR 2 TIu31ug-IIeuuod "5 СТ ттиовитаон “5 FT ваводовадпо9 '5 eT үнэд а Nt злоттопва *$ 11 в ива 28 291118 от виәтатоәр "8 6 eugdepoiuoQ— —— $ ОН jg V» 8 доловш "с в ~ 8113309125 "8 4 урившиоол) "5 9 ДУ '5 6 ъзвлодо "5 7 224213 “5 © єивоүецү5 “5 2 SS чволцәшо4 "5 1 ~ кешире ины ча eruepeullo3sng-:—-*- ~ rE NASTI е; Pc ч №. нео ЧИГ и. х zs К је 9,4 “іы 1 М - 6 ~ ~ VOR Пр 772. A NE a- 2-4 м В ~ ~ 5, > ot ь% ~ ` х 26 -31. 4. М 06 Ж У, -— E sim s 2 “а <> EY -. NC ot 1 -° мо. ~ [Vor. 15 352 ANNALS OF THE MISSOURI BOTANICAL GARDEN Museum of Natural History; MBG = Herbarium of the Mis- souri Botanical Garden. TAXONOMY Stemmadenia Benth. Bot. Voy. Sulph. 124. t. 44. 1844; Lindl. Veg. Кіпра. 601. 1847; Walp. Rep. 468. 1847; Рей. Nom. Bot. 2: 1270. 1874; Benth. & Hook. Gen. Pl. 2: 707. 1876; Miers, Apoc. S. Am. 74. 1878; Hemsl. Biol. Cent.-Am. Bot. 2: 310. 1881; Durand, Ind. Gen. Phan. n. 4615. 1888; Baill. Hist. Pl. 10: 196. 1891; K. Sch. in Engl. & Prantl, Nat. Pflanzenfam. 4: 148. 1895; Standl. Contr. U. S. Nat. Herb. 23: 1155. 1924. Odontostigma A. Rich. (non Zoll. & Mor.) Fl. Cub. Fanerog. 2: 86. 1853. Stemmaderia B. D. Jackson, Ind. Kew. 2: 331. 1894. err. typ. Lactescent shrubs or small trees 2-15 m. tall. Leaves entire, opposite, glabrous or pubescent, petiolate, the sheaths of the petioles meeting in a shallow ring about the stem and sheltering in the стих many small fusiform glands. Inflorescence a ter- minal reduced raceme of several flowers. Corolla large, in- fundibuliform or salverform, white or yellow, the limb of 5 equal lobes dextrorsely reflexed and occasionally auriculate, bearing 5 linear interior appendiculate folds opposite and slightly above the attachment of the stamens. Calyx 5-parted, the lobes imbricate, unequal, usually 3 larger interior and 2 smaller exterior, bearing several cycles of small fusiform glands within and near the attachment of the disc. Stamens 5, included, attached to the corolla at the summit of the proper-tube, alter- nate with the corolla-lobes; filaments very short and thick, unguiculate at the attachment to the anthers; anthers of 2 elongate unappendaged sporangia. Carpels 2, sessile, unilocu- lar, bearing many ovules upon a lateral binate ventral placenta, produced apically into a long filiform style; stigma terminal, borne upon a fleshy truncate clavuncle. Disc proper shallow, immersed, entire; nectaries fleshy, coalesced into a more or less irregular ring about, and slightly adnate to, the carpels. Fruit a pair of divaricate, leathery, glandular-punctate follicles con- taining many striate, albuminous, ecomose seeds immersed in an oily arilar pulp; embryo straight. 1928] WOODSON—STUDIES IN APOCYNACEAE, II 353 Type species: S. glabra Benth. Bot. Voy. Sulph. 124. 1. 44. 1844 SYNOPSIS OF THE SUBGENERA AND SECTIONS KEY TO THE SUBGENERA Corolla infundibuliform, the lobes dextrorsely reflexed, but only slightly auriculate; bracts immediately subtending the calyx. .................. а ра И nan ESTIS Subgen. I. Se Corolla salverform, the lobes dextrorsely reflexed and very strongly au late; bracts placed about midway upon the pedicels. .Subgen. П. хэрэг МОРА SUBGENUS I. EusrTEMMADENIA Woodson Subgenus I. Еовтеммаремл Woodson, n. subgen. Corolla infundibuliform, the lobes dextrorsely reflexed and very slightly auriculate; calyx-squamellae in several series of different lengths; sporangia of the anthers divergent at the base; rim of the coalesced nectaries irregularly folded and lobed; corollar appendages relatively long, 1.5-2.0 cm. long; bracts immediately subtending the calyx Section 1. Овоултав Woodson. Proper-throat of the corolla conical, about as long as broad. Ккү то THE SPECIES a. Calyx relatively short, 1-5 mm. long b. Calyx-lobes oblong to ovate, acute at the apex, 3-5 mm. long. c. Under-surface of leaves persistently and uniformly Ee e 6,5854 0. ИЕ ОЎ он о. ос . 8. tomentosa се. адалды of leaves slightly barbate in the axils of K mid- , becoming glabrous or glabrate....1a. S. tomentosa var. Palmeri bb. сейіс subreniform, rounded at Ше apex, 1 1-2 mm. long.. .2. S. sinaloana aa. Calyx relatively long, 1.5-3 cm. lon b. Proper-tube about as long as the calyx; inflorescence glabrous. ..3. S. glabra bb. Proper-tube much surpassing the calyx; inflorescence pubescent c. Upper-surface of leaves шаһгайе......................... . 8. obovata cc. Upper-surface of leaves persistently pubescent...4a. S. бе var. mollis 1. Stemmadenia tomentosa Greenm. Proc. Am. Acad. 35: 310. 1900; Standl. Contr. U. S. Nat. Herb. 23: 1156. 1924. Shrubs or small trees, 2-12 m. tall; leaves 8-15 cm. long, 5-7 em. broad, glabrous or glabrate above, tomentose beneath, petioles 4-8 mm. long; inflorescence 2-5-flowered; corolla yel- low, Ше proper-tube 2-2.5 em. long, the proper-throat conical, [Уог. 15 354 ANNALS OF THE MISSOURI BOTANICAL GARDEN 2-3 cm. long, 1.5-2 ст. broad at the orifice, the limb 2-3 em. broad; calyx-lobes 4-5 mm. long, the segments oblong to ovate, acute at the apex, somewhat imbricate, both the larger and the smaller yellowish; follicles 4-4.5 cm. long, 3-3.5 ст. broad, acute at the apex. Distribution: waste-lands, central and southern Mexico. Specimens examined: ‘ Mexico: Vera Cruz: San Juan, 1889, Heilprin & Baker (ANSP). JALISCO: lava beds near Zapotlan, May 19, 1893, Pringle 4870 (С түре, NY, US, MBG). SINALOA: Sinaloa, April 2, 1910, Rose, Standley & Russell 18874 (05). la. Var. Рашеп (Rose) Woodson, п. comb. Stemmadenia Palmeri ‘ Kose.” ex Urbina, Pl. Mex. 214. 1897, nomen. Slemmadenia Palmeri Rose ex Greenm. Proc. Am. Acad. 35: 311. 1900. " Stemmadenia Palmeri Rose & Standl.” in Standl. Contr. U. S. Nat. Herb. 23: 1156. 1924. Leaves glabrous or glabrate above, beneath barbate in the axils of Ше midvein, or glabrate; calyx-lobes 3-5 mm. long, greenish. Distribution: waste-land, and hedgerows, general over central and southern Mexico. Specimens examined: Мехтсо: CHIHUAHUA: Tierras Verdes, Мау, 1891, Hartmann 534 (G); southwestern Chihuahua, Aug.—Nov. 1885, E. Palmer М (С, US); Batopillas, April, 1892, Hartmann 1032 (С). SINALOA: Ymala, Aug. 16-25, 1891, E. Palmer 1470 (US TYPE); Mazotlan, April 5, 1910, Rose, Standley & Russell 14064 (US); vicinity of Rosario, April 14, 1910, Rose, Standley & Russell 14544 (US); San Ignacio, June 19, 1918, Montes & Salazar 405 (US); Guadaloupe, April 18, 1910, Rose, Standley & Russell 14675 (US); La Cruz, 1921, Ortega 4175 (US); between Rosario and Concepcion, July 27, 1897, Rose 8260 (US); San 1928 WOODSON—STUDIES IN APOCYNACEAE. 11 355 Ignacio, March 12, 1918, Montes & Salazar 268 (US); Rosario, July 8, 1897, Rose 1573 (US); Colomas, July 16, 1897, Rose 1688 (US). Jauisco: Вагапса, near Guadalajara, June, 1886, Е. Palmer 132 (G, US); Chiquilistlan, May 15, 1892, Jones 335 (MBG, US); Tequila, July 5-6, 1899, Rose & Hough 4777 (US); Baranca, near Guadalajara, May 28, 1891, Pringle 5151 (G); vicinity of Colima, April 5, 1897, Seler 3436 (G); Baranca of Guadala- jara, alt. 4000 ft., June 10, 1898, Pringle 6872 (G, NY, F, ANSP, US, MBG); Guadalajara, June 25, 1892, Pringle 5363 (G); San Sebastien, Jan. 15, 1927, Mexia 1490 (US); Bolanos Aug. 10-19, 1897, Rose 2888 (US). Durango: Chocala, March 7, 1899, Goldman 358 (US). MonELos: Cuernavaca, Мау 11, 1898, Pringle 6847 (US). Nayarit: Ojos de Agua, near Ixtlan, Sept. 23, 1926, Mexia 733 (US). Popular names of this variety are “Веггагсо,” “ Вегтасо 0 Tapaco,” and the gum of the fruit is said to be used like chicle (Montes & Salazar 405, US). The embarrassment of monographers who find themselves forced to regard as "typical" an anomalous form of a species because of priority in publication over a more common variety is illustrated in a peculiar fashion by Stemmadenia tomentosa Greenm. and its var. Palmeri. Аз early as 1891 the herbarium name “беттадета Palmeri Rose" was distributed with speci- mens of the glabrescent or barbate variety of the former species. The name did not appear in publication, however, until 1893, when Urbina, in compiling his *Catalogue of Mexican Plants,' happened upon specimens of the genus bearing the inscription of S. Palmeri Rose in a rather poor script, and erroneously published the name for the first time as a nomen nudum. Ur- bina mistook the name of Dr. Rcse for an abbreviation, and gave the author аз ‘‘Kosc.” It is indeed fortunate that a de- scription was not included under that authorship. In 1900 Dr. Greenman published Stemmadenia tomentosa, and in so doing spoke of the characteristics of 8. Palmeri Rose, which he evidently assumed to be a correctly published name. It was not until 1924 that Stemmadenia Palmeri was published (Vou. 15 356 ANNALS OF THE MISSOURI BOTANICAL GARDEN by Rose in Standley’s ‘Trees and Shrubs of Mexico.’ The legal place of publication of the species must evidently be regarded as ex Greenman, Proc. Am. Acad. 35: 311. 1900. 2. Stemmadenia sinaloana Woodson, п. sp.! Pl. 48, fig. 1. Shrubs or small trees; leaves 8-12 cm. long, 5-6 ст. broad, glabrous, or very slightly puberulent upon the lower surface, petiolate, the petioles 7-10 mm. long; inflorescence 1-4-Ноугегей; corolla yellow, the proper-tube 1.5-2 ст. long, the proper-throat conical, about 1.5 em. long, about 1.5 em. broad at the orifice, the limb 1.5-2 em. long; calyx about one-sixteenth the length of the proper tube, the segments ovate-reniform, 1.2 mm. long, about 4 mm. broad, obtuse at the apex, or completely rounded, scarcely imbricated, unequal, greenish; follicles unknown. Distribution: known only from the type locality in Sinaloa. Specimens examined: Mexico: SINALOA: Rosario, Jan. 1895, Lamb 467 (С ТУРЕ). 5. sinaloana is especially noteworthy in the genus Stemmadenia by reason of its peculiarly reduced calyx. In that respect it is closest related to 5. tomentosa Greenm., from which it differs in having a calyx less than one-half as large (1-2 mm. long), and in having the calyx-lobes subreniform and rounded at the apex instead of oblong and ovate with acute or acuminate apex as in the latter species. 3. Stemmadenia glabra Benth. Bot. Voy. Sulph. 124. 4. 44. 1844; Hemsl. Biol. Cent.-Am. Bot. 2: 310. 1881; Miers, Apoc. S. Am. 74. 1878; K. Sch. in Engl. & Prantl, Nat. Pflanzen- fam. 42: 149. 1895; Standl. Contr. U. S. Nat. Herb. 23: 1156. 1925; Standl. & Calderón, Lista Prélim. Pl. Sal. 174. 1925. Pl. 47, fig. 1. Shrubs or small trees, 2-10 m. tall; leaves 14-20 cm. long, 7-8 ст. broad, glabrous, petiolate, petioles 5-10 mm. long; ! Stemmadenia sinaloana sp. nov., arborea glabra vel subpuberulenta; foliis ob- longo-lanceolatis 8-12 сш. longis 5-6 cm. latis; petiolis 7-10 mm. longis; corollae tubo conico-infundibuliformo 3-3.4 em. longo, lobis ca. 1.5 cm. longis; calycis lobis parvis ovato-reniformibus inaequalibus ca. 2 mm. longis ca. 4 mm. latis obtusis, viridibus.—Sinaloa, Rosario, Jan. 1895, Е. H. Lamb 467 (Gray Herb., ТУРЕ). 1928 WOODSON-—STUDIES IN APOCYNACEAE. II 357 inflorescence 1-4-flowered; corolla deep yellow, the proper-tube 2-2.5 ст. long, the proper-throat conical, about 2 ст. long, 2-2.5 cm. broad at the orifice, the limb 2.5-3 ст. long; calyx about equalling the length of the proper tube, the segments 1.5- 2.5 сіп. long, .8-1.0 ста. broad, strongly imbricate іп two unequal series, the larger yellow, the smaller greenish yellow; follicles about 5 cm. long, 3-3.5 em. broad. Distribution: tropical forests and thickets, Central хана. Reported also from Mexico. Specimens examined: Costa Rica: between San Pedro de Montes de Oca and Cur- ridabat, Dept. San José, Feb. 2, 1924, Standley 32798 (US); Cartago, Feb. 1924, Standley 35459 (US). Honpuras: Amapala, Isla de Tigre, Feb. 14, 1922, Standley 20718 (US). Ет, Satvapor: vicinity of La Unión, Dept. La Unión, alt. 150 m., Feb. 13-21, 1922, Standley 20686 (С, NY, US); Laguna de Magugüe, Dept. La Unión, alt. 60 m., Feb. 18, 1922, Stand- ley 20948 (G, NY, US); La Unión, Sept. 21, 1860, PORUM (G). NICARAGUA: southwestern slopes of Santiago Volcano, near Masaya, alt. 300-480 m., July 5, 1923, Mazon 7647 (С, US); Ometepe Island, Jan. 1893, C. L. Smith (G); Managua, shores of Lake Managua, June 24, 1923, Mazon, Harvey & Valentine 7270 (US); Managua, vicinity, June 30, 1923, Mazon, Harvey & Valentine 7589 (US); Laguna de Masaya, July 6, 1923, Mazon 7727 (US). Dr. Sutton Hayes remarks (Sutton-Hayes, G) that the popular name of this species in El Salvador 18 ‘‘Cajon del Mico.” Ас- cording to Standley (Standley 32793, US), the popular name in Costa Rica is “Биеуоз de Caballo," or ''Girijarro," and the sap is used for corns and tooth-ache. 4. Stemmadenia obovata (Hook. & Arn.) K. Sch. in Engl. & Prantl, Nat. Pflanzenfam. 42: 149. 1895. Bignonia (?) obovata Hook. & Arn. Bot. Beech. Voy. 439. 1841. Stemmadenia pubescens Benth. Bot. Voy. Sulph. 125. 1844; (Vou. 15 358 ANNALS OF THE MISSOURI BOTANICAL GARDEN Miers, Арос. 5. Am. 74. 1878; Hemsl. Biol. Cent.-Am. Bot. 2: 310. 1881. Shrubs or small trees, 2-15 т. tall; leaves 10-20 cm. long, 7-10 cm. broad, pubescent, or glabrate above, petiolate, peti- oles 5-8 mm. long; inflorescence 1—6-flowered; corolla deep yel- low, the proper-tube 1.5-2.5 ст. long, the proper-throat 1.5-3 ст. long, 2-2.5 ст. broad at the orifice, the limb 1.5-2.5 cm. long; calyx much surpassed by the length of the proper-tube, the segments 1.5-2 cm. long, .8-1.0 ст. broad, strongly imbri- cated in two unequal series, both series yellowish; follicles 4- 4.5 ст. long, 3-3.5 cm. broad, acute at the apex. Distribution: tropical forests and thickets, southern Mexico and Central America. Specimens examined: Mexico: GUERRERO: El Согтејо, alt. 900 m., Мау 18, 1899, Langlassé 1029 (G). Costa Rica: Salinas, July, 1890, Pittier 1177 (US). Nicaragua: Managua, vicinity, June 30, 1923, Mazon, Har- vey & Valentine 7542 (US); La Paz, Dept. Leon, Jan. 31, 1903, Baker 2270 (G, US); Managua, June 30, 1926, Chaves 215 (US). Er SALVADOR: near La Cebadilla, 1922, Calderón 1280 (G); Laguna de Olomega, Dept. San Miguel, alt. 75 m., Feb. 20, 1922, Standley 21034 (G). 4a. Var. mollis (Benth.) Woodson, n. comb. Stemmadenia mollis Benth. Bot. Voy. Sulph. 125. 1844; Hemsl. Biol. Cent.-Am. Bot. 2: 310. 1881; Miers, Apoc. S. Am. 75. 1878; K. Sch. in Engl. & Prantl, Nat. Pflanzenfam. 4*: 149. 1895; Urbina, Pl. Mex. 214. 1897; Donn.-Sm. Enum. Pl. Guat. 4: 105. 1895; Areschoug, Pl. ca. Guayaquil Coll. 127. 1910; Standl. Contr. U. S. Nat. Herb. 23: 1156. 1924; Standl. & Calderón, Lista Prélim. Pl. Sal. 174. 1925. Stemmadenia calycina Brandg. Univ. Cal. Publ. Bot. 10: 188. 1922. Upper surface of leaves persistently tomentose. Distribution: tropical forests and hedgerows, southern Mex- ico, northern Central America, and northwest-central South America. 1928] WOODSON—STUDIES IN APOCYNACEAE. II 359 Specimens examined: Мех!со: Vera Cruz: Baños del Carrizal, Aug. 1912, Ригриз 6230 (а, NY, US, MBG); San Francisco, Мау, 1894, С. Г. Smith 1339-1374 (G); Remulatero, April, 1922, Purpus 8771 (G, NY, US, MBG). GUERRERO: Iguala, Aug. 1905, Rose, Painter & Rose 9274 (MBG, NY, US); El Correjo, May 18, 1899, Langlassé 1029 (US). Oaxaca: Camino де Tonomeca, Мау 7, 1917, Conzatli & Reko 3258 (US, MBG). СнтАРАЗ: Petapa, Мау 29, 1904, Goldman 1027 (US). Costa Rica: Salinas, July, 1890, Pittier 2908 (US); Nicoya, April, 1900, T'onduz 13900 (G, NY, US); Liberia, Dept. Guana- caste, April, 1893, Shannon 5042 (US); Las Huacas, Nicoya Peninsula, May 24, 1903, Cook & Doyle 723 (US). Ет, SALVADOR: Sonsonate, Dept. Sonsonate, alt. 220-300 m., March 18-27, 1922, Standley 22372 (G, NY, US); Laguna de Olomega, Dept. San Miguel, Feb. 20, 1922, Standley 21034 (US); La Cebadilla, Dept. San Salvador, 1922, Calderon 1230 (US); between San Martin and Laguna de Ilopanga, Dept. San Salvador, April 1, 1922, Standley 22539 (US). Nicaragua: Momotombo, May 27, 1895, С. L. Smith 126 (G, NY); Los Braziles, Jan. 28, 1928, Mell 28 (NY); south of Managua, March 3, 1922, Greenman & Greenman 5718 (MBG). GUATEMALA: Fiscal, alt. 3700 ft., May 31, 1909, Deam 6070 (G, US); Agua Caliente, March 28, 1922, Greenman & Green- man 5920 (MBG); Barranquillo, Dept. El Progreso, May 21, 1920, Popenoe 977 (US); between Chiquín and Crapeche Grande, Dept. Guatemala, March 19, 1905, Pittier 188 (US); El Rancho, Dept. Jalapa, April 4, 1905, Maxon & Hay 8766 (US); Dept. Jalapa, March 10, 1905, Kellerman 4511 (US). Ecuapor: Guayaquil, Feb. 1885, Rusby 931 (NY); hillsides near Guayaquil, Sept.-Oct. 1925, Mille 59 (NY); Durán, Nov. 5-8, 1918, Rose & Rose 23612 (NY). Вошута: near Yungas, alt. 4000 ft., 1885, Rusby 1168 (NY). Section 2. Сагвотпав Woodson. Proper-throat of the cor- olla cylindrical, much longer than broad. (Vor. 15 360 ANNALS OF THE MISSOURI BOTANICAL GARDEN Кет то THE SPECIES a. Proper-tube about equalling the length of the proper-throat; corolla-tube, sensu-latiore, 3-3.5 cm. long. b. Calyx-lobes 1.5-2 mm. long; corolla-limb 5-8 mm. broad..... 6. S. Alfart bb. Calyx-lobes 5-7 mm. long; corolla-limb 10-15 mm. broad. .6. S. Greenmanii аа. Proper-tube much surpassed by the length of the proper-throat; corolla- tube, sensu-latiore, 4.5-6 cm. long. Calyx 1-1.5 cm. long, the lobes distinctly imbricated. . . . . 7. S. Galeottiana bb. Calyx 4-5 mm. long, the lobes scarcely imbricated....... 8. S. macrophylla 5. Stemmadenia Alfari (Donn.-Sm.) Woodson, n. comb. Tabernaemontana Alfari Donn.-Sm. Bot. Gaz 24: 396. 1897. Small tree 3-4 m. tall; leaves 7-11 спа. long, 3.5-5 ст. broad, glabrous, acuminate, subspathulate, petiolate, petioles 1-1.5 em. long; inflorescence 1-3-flowered; corolla infundibuliform or oc- casionally subinfundibuliform, yellow or yellowish white, the tube, sensu-latiore, 3-3.5 ст. long, the limb 1-1.5 cm. broad; calyx-lobes 1.5-2 mm. long, 1.5-2 mm. broad, scarcely imbri- cated in two series, both the inner and the outer yellowish; follicles unknown. Distribution: hedgerows and waste-lands, Costa Rica. Specimens examined: Costa Rica: San Pedro, near San Ramón, hedgerows, alt. 1300 m., April 13, 1918, Топдиг 17653 (Е); Limoncito and Vuelta, alt. 1100 m., March, 1897, Pittier 11094 (US TYPE). 6. Stemmadenia Greenmanii Woodson, n. sp.! Pl. 48, fig. 2. Shrubs or small trees 1-6 m. tall; leaves 8-12 cm. long, 4—5 ст. broad, glabrous, petiolate, petioles 5-8 mm. long; inflores- сепсе 2—5-flowered ; corolla yellowish white, the proper-tube about 1.5 ст. long, the proper-throat cylindrical, about 2.0 cm. long, about .8 ст. broad at the orifice, the limb 1-1.5 спа. broad; calyx about one-third the length of the proper-tube, the seg- ments .5-.7 cm. long, .3-.4 cm. broad, strongly imbricated in two unequal series, both series yellowish; immature specimens oblong-lanceolate, acute at the apex. 1Stemmadenia Greenmanii sp. nov., arborea glabra; foliis oblongo-lanceolatis 8-12 сш. longis 4-5 cm. latis; petiolis 5-8 mm. longis; corollae tubo cylindrico- infundibuliformo 3-5 cm. longo, lobis 1-1.5 cm. longis; lobis calycis ovatis inae- qualibus ca. .5 cm. longis 3-4 mm. latis flavis; folliculis oblongo-lanceolatis acuti- busque.—Costa Rica, San Ramon, June 4, 1901, Brenes 14275 (Gray Herb., ТУРЕ). 1928] WOODSON—STUDIES IN APOCYNACEAE. II 361 Distribution: tropical forests and thickets, Costa Rica. Specimens examined: Costa Rica: San Ramón, alt. 1100 m., Мау 29, 1901, Brenes 14275 (С TYPE); San Ramón, June 4, 1901, Brenes 14278 (С). This species is evidently very local, but is very distinct. The nearest related species is undoubtedly S. Alfari, from which, however, it differs radically in the size of the calyx and all the floral parts. The species is dedicated to Dr. J. M. Greenman, by all odds the most discriminating of recent students of the group. 7. Stemmadenia Galeottiana (A. Rich.) Miers, Арос. 5. Am. 76. 1878. Pl. 47, figs. 2-3. Odontostigma Galeottiana A. Rich. in Sagra, Hist. Cub. 11: 868. t. 60 (Fl. Cub. Fanerog. 2: 86). 1853; Walp. Ann. 5: 477. 1858. Echites bignoniaeflora Schl. Linnaea 26: 372. 1853. Stemmadenia bignoniaeflora (Schl.) Miers, Арос. 5. Am. 76. 1878; Donn.-Sm. Enum. Pl. Guat. 5: 51. 1899; Standl. Contr. 0. S. Nat. Herb. 23: 1156. 1924. Stemmadenia insignis Miers, Apoc. S. Am. 76. t. 10В. 1878; Hemsl. Biol. Cent.-Am. Bot. 2: 310. 1881; Standl. Contr. U. S. Nat. Herb. 23: 1156. 1924. Tabernaemontana laurifolia Schott (non L., nec Ker, neque Blaneo) ex Miers. Apoc. S. Am. 76. 1878 momen. Stemmadenia bella Miers, Apoc. S. Am. 77. 1878; Donn.- Sm. Enum. Pl. Guat. 5: 51. 1899; Standl. Contr. U. 5. Nat. Herb. 23: 1156. 1924. Stemmaderia Galeotiianum В. D. Jackson, Ind. Kew. 2: 331. 1844, err. typ. Shrubs, 1-3 m. tall; leaves 9-12 cm. long, 4-5 ст. broad, glabrous or slightly puberulent upon the lower surface, petio- late, petioles 8-11 mm. long; inflorescence 1—4-flowered; corolla yellow, the proper-throat 4-5 cm. long, 1.0-1.3 em. broad at the orifice, the proper-tube 8-10 mm. long, the limb 2.5-3 cm. long; calyx segments 10-14 mm. long, 4-7 mm. broad, strongly imbricated in two unequal series; follicles 2-2.5 ст. long, 1.5-1.7 ста. broad. [Уох. 15 362 ANNALS OF THE MISSOURI BOTANICAL GARDEN Distribution: tropical forests of southern Mexico and Costa Rica. Specimens examined: МЕх!со: Vera Cruz: Teocelo, Мау 8, 1901, Goldman 575 (US); Zacua- pan, March, 1917, Purpus 7740 (G, NY, US, MBG); Orizaba, March 23, 1867, Bilimek 269 (С); Orizaba, date lacking, Botteri 988 (G); Textolo, alt. 3500 ft., April 26, 1899, Pringle 8103 (G, NY, ANSP, US, F, MBG); Orizaba, April, 1866, Bourgeau 2440 (G, US). Oaxaca: exact locality lacking, 1841, Galeotti 1605 (NY co- TYPE?) ; Sontecomopan, Galeotti 1599 (US). Yucatan: Merida, April 14, 1887, Millspaugh 27 (Е); Izamal, cultivated for its flowers, date lacking, Саитег 23204 (Е); Merida, Quinta del Obispo, March 18, 1865, Schott 430 (US, F). Since Odontostigma Galeotiiana A. Rich. and Echites bignoniae- flora Schl. were both published in 1853 at unknown months, according to the fly-leaves of the journals in which they appeared, it was perplexing whether to conserve Stemmadenia Galeottiana or S. bignoniaeflora. However, on the fly-leaf of Asa Gray’s copy of ‘Linnaea’ 26, in which E. bignoniaeflora was published, appears the note in Dr. Gray’s handwriting that the publication did not actually leave the press until August, 1854. In the absence of contradiction, then, it is assumed that Richard’s species appeared in 1853, and is therefore considered to have priority. 8. Stemmadenia macrophylla Greenm. Proc. Am. Acad. 35: 310. 1900; Donn.-Sm. Enum. Pl. Guat. 6: 83. 1903. Shrubs or small trees; leaves 15-20 cm. long, 5-7 ста. broad, glabrous, petiolate, petioles 1.5-2 ст. long; inflorescence 1-4- flowered; corolla yellow, the proper-throat cylindrical, 2.5-3 cm. long, .8-1.2 ст. broad, the proper-tube 1.5-2 ст. long, the limb 2.5-3 cm. broad; calyx about one-half the length of the proper- tube, the segments 4—6 mm. long, 3-4 mm. broad, scarcely imbricated in two unequal series, both series yellowish; follicles unknown. Distribution: tropical thickets of Guatemala. Specimens examined: 1928] WOODSON—STUDIES IN APOCYNACEAE. II 363 GUATEMALA: Pansamalá, Dept. Alta Verapaz, alt. 8800 ft., Jan. 1886, T'uerckheim 981 (С түре, NY, US, MBG); Coban, Dept. Alta Verapaz, April, 1889, Donnell-Smith 1800 (US); San Carlos Miramar, March 19, 1921, 750 m., Tonduz & Rojas 147 (MBG). SuBGENUS П. OcHRoDAPHNE Woodson Subgenus I. Оснкорлрнче Woodson, n. subgen. Corolla salverform, the lobes dextrorsely reflexed and strongly auriculate; calyx-squamellae in two or three series of nearly uniform length; sporangia of the anthers nearly parallel to the base; rim of the coalesced nectaries nearly smooth; corollar appendages relatively short, 5-7 mm. long; bracts placed mid- way upon the pedicels. Name coined from Фур$с, yellow, and 5афут, laurel, from the popular name of Stemmadenia grandiflora (Jacq.) Miers, “Yellow Laurel." Кеу TO THE SPECIES a. Calyx less than one-half the length of the corolla-tube. b. Bracts scarious; leaves ovate to ovate-oblong. с. Inner series of calyx-lobes only slightly longer than the outer; leaves glabrous throughout, glaucous benea d. Inflorescence several- or many-flowered; Зани ав broad ав Ше length of the tube. e. СУТ 4—6 mm. long, yellowish, appressed; leaves м КОРЕН 2 гала» 9. decipiens ее. буглаа 10-15 mm. long, green, spreading; leaves ovate-oblong. 2,202 502... 10. 5. grandiflora 44. 82 l-flowered by abortion; corolla-limb about e-half as broad as the length of the tube. .11. S. pauciflora ec. Inner de of calyx-lobes about twice as long as the outer; finely rufose-puberulent beneath.......... 12. S. Pennellit bb. Bracts foliaceous; leaves lanceolate.................. 13. S. eubracteata a&. Calyx equalling or nearly equalling the length of the corolla-tube. b. Calyx-lobes tinea рови leaves oblong-lanceolate, glabrous Штоц һош&......... еа; 14. S. Robinsonii bb. Calyx-lobes ovate; leaves spatulate, glabrous above, the under- surface халан barbate in the axils of the midvein. ГГ” 28 15. 8. Donnell-Smithii 9. Stemmadenia decipiens Woodson, n. sp.! 1 Stemmadenia decipiens sp. nov., arborea glabra; foliis ovatis 7-10 em. longis 4-7 cm. latis, petiolis 5-7 mm. longis; corollae tubo salverformo 2-3 ст. longo, lobis 1-2 сш. longis; lobis calycis ovatis inaequalibus 4-6 mm. longis 2-3 mm. latis flavis; folliculis ovato-oblongis acutibusque.—Mexico, between Rosario and Colomas, Sinaloa, July 12, 1897, J. N. Rose 1614 (US. TYPE). Vor. 15 364 ANNALS OF THE MISSOURI BOTANICAL GARDEN Shrubs or small trees, 2-10 m. high; leaves 7-10 cm. long, 4-7 cm. broad, glabrous, petiolate, petioles 5-7 mm. long; in- florescence 3-9-flowered; corolla yellow or yellowish white, the tube 2-3 cm. long, 4-5 mm. broad at the orifice, the limb 1-2 cm. broad; calyx about one-fifth the length of the tube, the segments 4—6 mm. long, 2-3 mm. broad, slightly imbricated in two unequal series, both series yellowish, appressed; imma- ture follicles ovate, attenuate at the apex, mature follicles un- known. Distribution: southern Mexico and adjacent Central America. Specimens examined: MExico: SINALOA: between Rosario and Colomas, July 12, 1897, Козе 1614 (US, Мо. 300461 түре, 300462); near Rosario, July 24, 1897, Rose (US). Oaxaca: Pochutla, April 19, 1917, Conzatti, Reko & Makrinius 3172 (МВО). Nicaracua: Managua, 1925, René 78 (MBG). This species has been called decipiens because it possesses the smallest calyx of the subgenus Ochrodaphne, thus recalling the small calyx-lobes of 8. Palmeri in the subgenus Hustemma- denia, for which it has been mistaken. 10. Stemmadenia grandiflora (Jacq.) Miers, Арос. 5. Am. 75. 1878. Pl. 47, fig. 4. Tabernaemontana grandiflora Jacq. Enum. Pl. Carib. 14. 1762; L. Mant. 53. 1767; Willd. Sp. Pl. 12: 1245. 1798; Lam. Dict. 7: 528. 1806; Roem. & Schult. Syst. 4: 428. 1819; А. ОС. in DC. Ргодг. 8: 368. 1844; С. Поп, Gen. Syst. 4: 88. 1887; Sesse & Мосіпо, Fl. Мех. 431. 1894; Ramirez, Pl. Мех. 155. 1902, Pulle, Enum. Vasc. Pl. Зиг. 381. 1906; Standl. Contr. U. S. Nat. Herb. 27: 308. 1928. Shrubs or small trees; leaves 6-8 cm. long, 3-5 ст. broad, glabrous, petiolate, petioles 5-7 mm. long; inflorescence 2-9- flowered; corolla yellowish white, the tube 3-3.5 cm. long, 4—5 mm. broad at the orifice, the limb 1.5-2 cm. broad; calyx about one-third the length of the tube, the segments 8-12 mm. broad, 10-15 mm. long, closely imbricated in two unequal series, both 1928] WOODSON—STUDIES IN APOCYNACEAE. II 365 series green, spreading; follicles 3-3.5 ст. long, 2-3 cm. broad, acute at the apex. Distribution: tropical forests, southern Mexico, Central Amer- ica, and northeastern South America. Specimens examined: Mexico: SrNALOA: Colomas, July 16, 1897, Rose 1711 (US); near Colomas, July 14-17, Rose (US). NAYARIT: vicinity of Acaponeta, Теріс, April 12, 1910, Rose, Standley & Russell 14484 (US). Costa Rica: exact locality and date lacking, Топдиг 17653 (US). PANAMA: Chagres, Jan.-March, 1850, Fendler 234 (С, MBG); exact locality and date lacking, Duchassaing (G); Puerto Reme- dios, Chiriqui, March 31, 1911, Pittier 8888 (NY, US); Fato, Dept. Colon, along the beach, July 8-10, 1911, Рийет 3940 (US); David, Chiriqui, Feb. 25, 1911, Pittier 2824 (US); Cana, April 17, June 8, 1908, Williams 803 (US); Cerro Gordo, near Culebra, June 29, 1911, Pittier 3739 (US); Paso del Olá, Prov. Coclé, Dec. 7-9, 1911, Pittier 5011 (US); Mount Hope Ceme- tery, Canal Zone, Dec. 28, 1923, Standley 28840 (US); Punta Раша, Nov. 3, 1921, Heriberto 209 (US); Puerto Obaldia, forests, Oct. 11, 1911, Pittier 4406 (US); Sabana de Juan Corso, Prov. Panama, near Chepo, Sept. 1911, Рийег 4748 (US); Pan- ama City, old Experiment Station, June 13, 1923, Mazon, Har- vey & Valentine 7084 (US); Bella Vista, near Panama City, June 12, 1923, Mazon & Valentine 6948 (US); Juan Diaz, Prov. Panama, near Tapia River, June 1-3, 1923, Махоп & Harvey 6751 & 6646 (US); Corozal, Canal Zone, Aug. 1924, Stevens 90 (US); Chivi-Chivi Trail, 2 mi. above Red Tank, Canal Zone, May 28, 1923, Maxon & Harvey 6599 (US); Barro Colorado Is., Canal Zone, Aug. 18, 1927, Kenoyer 500 (US); Changuinola Valley, 1927, Cooper & Slater 63a (US); Taboga Is., Feb. 26, 1923, Macbride 2798 & 2799 (US); Barro Colorado Is., Canal Zone, Nov. 18-24, 1925, Standley 40994 (05). VENEZUELA: Tovar Colony, Aug. 16, 1855, Fendler 1087 (С, NY); San Martin, on the Rio de Palomar, Oct. 15, 1922, Рийет 10516 (G); between La Guaira and Rio Grande, June 12, 1917, [Vor. 15 366 ANNALS OF THE MISSOURI BOTANICAL GARDEN Curran & Haman 971 (G, US); San José & Rio Chico, June 16, 1913, Pittier 6856 (NY); Cierucunté, April 10, 1922, Pittier 10288 (NY); La Guavia, July 4, 1900, Robinson & Lyon (US); Puerto La Cruz, April, 1914, Jahn 336 (US); Rio Chico, Miranda, June 20, 1923, Jahn 1280 (US); between San José and Las Trincheras, Fed. Dist. (Caracas), Oct. 4, 1921, Pittier 11 (US); Curucuti, March 19, 1918, Piitier 7774 (US); Perijá, Zulia, 1917, Tejera 14 (US). Согомвтл: Turbaco, Nov. 1920, Heriberto 461 (US); Carta- gena, 1919, Heriberto 249 (US); between Ciénaga de Santa Marta and the foothills, June 22-30, 1906, Pittier 1594 (US, NY); San Martin de Loba and vicinity, Bolivar, April-May, 1916, Curran 12 (US); Santa Marta, 1898-1901, H. H. Smith 1639 (MBG, NY, ANSP, US). Потсн Guiana: Paramaribo, on way to Kwatta, Samuels (US); Paramaribo, forests on the way to the farm of Kwatta, April 27, 1916, Samuels 384 (G); Paramaribo, forest behind Gongrypstreet, April 12, 1916, Samuels 385 (G); Paramaribo, May 10, 1905, Mayo (ANSP); “Surinam,” Weigelt (ANSP). This common species is known popularly in Panama as “Huevo de Gato,” “ Lechosa,” and “‘ Venenillo’’; in Venezuela as ‘‘Hueves de Burro"; and in Mexico as ‘‘Lechoso.” Called “yellow laurel” by G. Don. 11. Stemmadenia pauciflora Woodson, п. sp.! Pl. 49, fig. 1. Shrubs ог small trees; leaves 8-12 cm. long, 2.5-5 ст. broad, glabrous, petiolate, petioles 2-3 mm. long; inflorescence 1-flow- ered by abortion; corolla yellow or yellowish white, salverform, the tube 3-4 cm. long, 3-4 mm. broad at the arifice, the limb 1-1.5 cm. broad; calyx about one-fourth the length of the tube, the segments 7-9 mm. long, 6-9 mm. broad, str ongly imbri- cated in two unequal series, both series green; follicles unknown. Distribution: north-central Colombia and Guiana. 'Stemmadenia pauciflora sp. nov., arborea glabra; foliis oblongo-lanceolatis 8-12 ст. longis 2.5-5 ст. latis, petiolis 2-3 mm. longis; cymis unifloris abortivis; corollae tubo salverformo 3-4 cm. longo, lobis 1-1.5 сіп. longis; lobis calycis ovatis inaequalibus 7-9 mm. longis 6-9 mm. latis viridis; folliculis ignotis.—Colombia, between Espinal and Cuamo, Tolima, open loam along stream, alt. 350-400 m July 21, 1917, Pennell & Rusby 186 (NY TYPE). 1928] WOODSON—STUDIES IN APOCYNACEAE, II 367 Specimens examined: CoLoMBIA: open loam along stream, between Espinal and Cuamo, alt. 350-400 m., Tolima, July 21, 1917, Pennell & Rusby 186 (NY TYPE). Потсн Салама: “in sylvis pr. urbem Paramaribo,” March- April, 1844, Kappler 1565 (MBG). Stemmadenia pauciflora, so-called because of the singularly reduced inflorescence, is equally distinct because of the short corolla-limb. At present the two specimens referable to the species constitute a rather scattering range, but doubtless with increased collecting activity additional localities will become known. The young inflorescence is normally composed of sev- eral buds, all of which abort very early except the one destined to produce the fully-developed flower. This character of the inflorescence is demonstrated nicely by the two above-cited specimens. Upon Kappler 1565 three inflorescences appear, one with one aborting and one developing bud, and two with full- blown flowers and one aborted bud each. Pennell & Rusby 186 likewise demonstrate this remarkable propensity. On that sheet (NY) three inflorescences appear, one with one aborting and one developing bud, another with two aborted and one developing bud, and another with a full-blown flower and one aborted bud. 12. Stemmadenia Pennellii Woodson, n. sp.! Shrubby vines (?) or shrubs; leaves 7-9 cm. long, 3-4 cm. broad, glabrous or glabrate above, beneath softly rufous-puberu- lent, petiolate, petioles 1-3 mm. long; inflorescence 2—4-flowered; corolla salverform, yellow, the tube 3-3.5 cm. long, the limb 2.5-3 cm. broad; calyx-lobes in two very unequal series, the inner about 2 cm. long, the outer 1.2-1.5 ст. long, 7-10 mm. broad, strongly imbricated; follicles unknown. 1Stemmadenia Pennellii sp. nov., arborea vel vinea frutescens (?), foliis ob- longo-lanceolatis 7-9 cm. longis 3—4 cm. latis supra glabris vel glabratis subtus rufo-puberulentis; petiolis 1-3 mm. longis; corollae tubo salverformo 3-3.5 cm. longo, lobis ca. 2.5 ст. longis; lobis calycis majusculis inaequalibus 2-serialibus, inferioris ca. 2 ст. longis superioris 1.2-1.5 cm. longis, virido-flavibus; folliculis ignotis.—Colombia, Turbaco, Bolivar. Shrubby vine, thin loam over white rock, alt. 150-200 m., March 27, 1918, Pennell 4755 (Gray Herb. TYPE). [Уоь. 15 368 ANNALS OF THE MISSOURI BOTANICAL GARDEN Distribution: southern Mexico and northern Colombia. Specimens examined: Мех!со: GUERRERO: Achatla, May, 1926, Reko 4892 (US). СотомвгА: Turbaco, Bolivar. Shrubby vine. Thin loam over white rock, alt. 150-200 m., March 27, 1918, Pennell 4755 (С түре, US, Е, MBG). This species, although closely related to S. grandiflora, is very distinctive, not only because of the strikingly unequal calyx-lobes, but because of the rufous puberulence of the leaves. The flowers, also, appear to be more turbinate in the reflexion of the corolla-lobes than any other species of Ochrodaphne, but this character has not been noted because most of the specimens of that subgenus are so poorly pressed, and Dr. Pennell’s are so carefully prepared, that use of this character would be danger- ous. The species is dedicated to Dr. Francis W. Pennell, of the Academy of Natural Sciences of Philadelphia, the collector of the type specimen. 13. Stemmadenia eubracteata Woodson, п. вр! РІ. 49, fig. 2. Shrubs or small trees; leaves 6-8 cm. long, 2-3 cm. broad, glabrous, petiolate, the petioles 4-5 mm. long; inflorescence 2-5- flowered; corolla salverform, yellow, the tube about 2.5 cm. long, the limb about 1.5 ст. broad; calyx-lobes 3-4 mm. broad, 8-10 mm. long, all green, spreading; bracts semifoliaceous; fol- licles unknown. Distribution: known only from the type locality in Guatemala. Specimens examined: GUATEMALA: Volcan Tecuamburro, Dept. Santa Rosa, alt. 2000 m., Feb. 1893, Heyde & Lux 4588 (С TYPE). This species is one of the most remarkable species of Ochro- daphne by reason of the spreading calyx, the narrow leaves, and above all the curiously foliaceous bracts, which are abso- lutely different from those of any other species of the genus Stemmadenia. ! Stemmadenia eubracteata sp. nov., arborea; foliis lanceolatis 6-8 cm. longis 2-3 cm. latis glabris, petiolis 4-5 mm. longis; corollae tubo salverformo 2.5 cm. longo, lobis ca. 1.5 cm. longis; lobis calycis 3-4 mm. latis 8-10 mm. longis; bracteis semifoliaceis; folliculis ignotis.—Guatemala, Volcan Tecuamburro, Dept. Santa Rosa, alt. 2000 m., Feb., 1893, Heyde & Luz 4538 (Gray Herb. TY»k). 1928] WOODSON—STUDIES IN APOCYNACEAE. II 369 14. Stemmadenia Robinsonii Woodson, п. sp.’ Shrubs or small trees; leaves 12-16 cm. long, 4-5 cm. broad, glabrous, petiolate, the petioles 1-3 mm. long; inflorescence 2-3-flowered; corolla salverform, yellow, the tube 2-2.5 cm. long, the limb about 1-1.5 em. broad; calyx-lobes linear-lanceo- late, 1.5-2 cm. long, 3-4 mm. broad, very slightly imbricated in two unequal series, both series yellow, appressed; follicles unknown. Distribution: known only from the type locality in Costa Rica. Specimens examined: Costa Rica: Talamanca Mts., March, 1894, Pittier 8617 (US TYPE). This species is dedicated to Dr. B. L. Robinson, who, in 1899, questioned its determination as S. bella Miers, and called atten- tion in a note upon the specimen to the peculiar calyx, which is the most striking characteristic of the species. In addition to the calyx, S. Robinsonii differs from its nearest relative, 8. Donnell-Smithii, in the leaves, which are glabrous and oblong- lanceolate in the former, and spatulate and barbate in the latter. 15. Stemmadenia Donnell-Smithii (Rose) Woodson, n. comb. Tabernaemontana Donnell-Smithii Rose, Bot. Gaz. 18: 206. 1893 Tabernaemontana Donnell-Smithii Rose var. costaricensis Donn.- Sm. Bot. Gaz. 24: 397. 1897. Shrubs or small trees; leaves 6-8 cm. long, 3-3.5 cm. broad, spatulate, minutely glandular-puberulent or glabrate above, be- neath conspicuously barbate in the axils of the midvein, petio- late, the petioles 1-2 mm. long; inflorescence 1—4-flowered; corolla yellow, salverform, the tube 2.5-3 cm. long, the limb 1.5-2 em. long; calyx nearly equalling the length of the corolla- tube, 2-2.5 cm. long, the lobes 1.5-2 cm. broad, in two closely imbricated yellowish series; follicles about 3.5 ст. long, about 3 em. broad, rounded at the apex. ! Stemmadenia Robinsonii sp. nov., arborea glabra; foliis oblongo-lanceolatis 12-16 cm. longis 4-5 cm. latis, petiolis 1-3 mm. longis; corollae tubo salverformo 2-2.5 cm. longo, lobis 1-1.5 cm. longis; lobis calycis linearo-lanceolatis 1.5-2 ст. longis 3-4 mm. latis; folliculis ignotis.—Costa Rica, Talamanca Mts., March, 1894, Pittier 8617 (US TYPE). [Vor. 15 370 ANNALS OF THE MISSOURI BOTANICAL GARDEN Distribution: tropical forests, southern Mexico and Central America. Specimens examined: Mexico: locality and date lacking, Gregg 893 (MBG). GUERRERO: La Correa, Oct. 5, 1898, Langlassé 427 (G, US). Costa Rica: Nicoya, April, 1900, Tonduz 13904 (G, NY); Santa Clara, Sept. 1896, Cooper 10241 (US); Мабатһй, N icoya Peninsula, May 23, 1903, Cook & Doyle 706 (US); Nicoya, alt. 200 m., May 22, 1903, Cook & Doyle 686 (US); Nicoya, forests, Топаиг 13904 (US); Саршт, on the Rio Grande de Taracales, Prov. Alojuela, April 2, 1924, Standley 40220 (US); Arenal, May 5, 1922, Valerio 86 (US). British Honpuras: Middlesex, Jan. 17, 1926, Ricard 13 (US). Honpuras: Ceiba, Aug. 20, 1916, Dyer A84 (US). NICARAGUA: Las Nubes, June 28, 1923, Maxon, Harvey & Valentine 7502 (US). GUATEMALA: St. Thomas, May 29, 1909, Deam 6052 (G); Escuintla, alt. 1100 ft., March, 1890, Donnell-Smith 2404 (G); San Felipe, Dept. Retalhulen, alt. 2050 ft., April, 1892, Donnell- Smith 2763 (С, US түре, NY, Е, MBG); Barranca de Eminen- cia, Dept. Amatitlan, alt. 1400 ft., Feb. 1892, Donnell-Smith 2762 (G, NY, MBG); Escoba, June 2, 1922, Standley 2462 (US); Hacienda el Baul, Dept. Escuintla, March 2, 1921, Ton- duz & Rojas 36 (US); Santa Lucia, Escuintla, March 2, 1905, Kellerman 5286 & 6275 (US); Mazatenango, border of forest, Feb. 19, 1905, Maxon & Hay 3490 (US); San Jose de Escuintla, April, 1892, Donnell-Smith 2765 (US); San Juan Mixtan, April, 1890, Donnell-Smith 2405 (US); Primavera, Dept. Sololá, Oct. 1891, Shannon 120 (US); Rio Toro Amarillo, Llanuras de Santa Clara, April, 1896, Donnell-Smith 6646 (US ТУРЕ var. costa- ricensis); Santa Barbara, Dept. Sololá, Aug. 1891, Shannon 162 (US); Naranjo, Dept. Escuintla, March, 1892, Donnell-Smith 2764 (US). Еш Satvapor: vicinity of San Salvador, alt. 650-850 m., Dec. 20, 1921-Jan. 4, 1922, Standley 19187 (US, С, NY); vicin- ity of Izaleo, Dept. Sonsonate, March 19-24, 1922, Standley 21865 (G, US); same locality and date, Standley 22218 (G, NY, US); vicinity of Ixtepeque, Dept. San Vicente, alt. 400 m., 1928] WOODSON-—STUDIES IN APOCYNACEAE. II 371 March 6, 1922, Standley 21463 (G, US); San Salvador, 1921, Calderon 187 (NY, US); San Salvador, 1900, Renson 106 (US); Armenia, Dept. Sonsonate, April 18, 1922, Standley 23452 (US); Dept. Ahuachapán, 1923, Padilla 331 (US); Izalco, Dept. Son- sonate, Feb. 17, 1907, Pittier 1936 (US). In referring this species to the genus Tabernaemontana, Dr. Rose! was fully cognizant of its affinities, and especially of its relation to Stemmadenia grandiflora, but preferred to assign it to the former genus, remarking “Т. grandiflora, as is known, was referred by Miers to Slemmadenia, but is retained by Mr. Hems- ley in Tabernaemontana. The difference between these two gen- era is sometimes a little difficult to determine." Dr. Rose further added an interesting note concerning the species: ‘‘Capt. Smith observes of this plant: ‘It is not exactly a tree in habit. It occurred everywhere as I went from the coast up to the slopes of the volcanoes at an elevation of 5,000 ft. Тһе natives call it Cobal (varnish gum).’ Other popular names for the species are ‘Coj6n’ and 'Cojón de puerco.’ ” The observations of Dr. Rose quoted above are representative of the attitude with which the majority of botanists have re- garded the genus Stemmadenia. This paper will be successful if merely it demonstrates the numerous precise differences be- tween the genera Tabernaemontana and Stemmadenia. EXCLUDED SPECIES Stemmadenia guatemalensis Müll.-Arg. Linnaea 30: 410. 1859. = Malouetia guatemalensis (Miill.-Arg.) Standl. Jour. Wash. Acad. Sci. 15: 459. 1925. (Malouetia panamensis Müll.-Arg. in Van Heurck, Pl. Nov. 185. 1871.) List or ExsiccATAE CITED The distribution numbers are printed in italics; collections distributed without numbers are indicated by a dash. The numbers in parentheses indicate the species numbers in the present revision. Baker, C. F. 2270 (4). Chaves, D. 215 (4). Bilimek, —, 269 (7). Conzatti, C. & Reko, B. P. 3258 (4a). Botteri, M. 988 (7). Conzatti, C., Reko, B. P. & Makrinius, Bourgeau, M. 2440 (7). M. 3172 (9 enes, А. M. 14275, 14278 (6). Cook, О. Е. & Doyle, C. B. 723 (4a); Calderón, S. 1230 (4); 187 (15). 120, 152, 706, 686 (15). 1 Rose, Bot. Gaz. 18: 206. 1893. 372 Соорег, С. Р. 10241 (15). Cooper, С. Р. & Slater, С. М. 63a (10). Сигтап, Н. М. 12 (10). Curran, Н. М. & Haman, 8. 971 (10). Пеаш, С. 6070 (4а); 6052 (15). Donnell-Smith, J. 1800 (7); 2404, 2406, 2762, 2763, 2764, 2765, 6646 (15). Duchasssing: E. P. —, (1 Fendler, A. 1027 (10). Galeotti, H. 1599, 1605 (7). Gaumer, С. Е. 23204 (7). Goldman, Е. А. pi (la); 1027 (4a); 76 (7). Gregg, J. 893 (15). Greenman, J. M. & Greenman, M. T. 5718, 5920 (4a). Hartmann, С. У. 684, 1032 (1а). Науев, 8. — ( Heilprin, А. & Bakar, C. F. — (1). Heriberto, Bro. 209, 461, 249 (10). Heyde, E. T. & Lux, —, 4538 (13). Jahn, A. 336, 1280 (10). Jones, М. Е. 336 (1а). Kappler, А. 1665 (11). Kellerman, У. А. 4511 (4a); 6286, 6275 (15). Кепоуег, L. А. 500 (10). Lamb, Е. Н. 467 (2). Tanglassé, E. 1029 (4); 427 (15). Macbride, J. Е. 2798, 2799 (10). Maxon, У. R. 7647, 7727 (3). Maxon, У. В. & Harvey, А. D. 6599, 6646, 6751 (10). Maxon, W. R., Harvey, A. D. & Val- entine, A. T. 7270, 7539 (3); 7542 (4); 6948, E 7602 (10). & Hay, В. 3766 (4a); (15 Maxon, W. R. & Valentine, A. T. 6948 (10). Mell, C. D. 28 (За). Mexia, Y. des: 1490 (1a). Mille, L. 59 (4a). Millspaugh, С. Е. 27 (7). Montes, ka N. & Salazar, А. Е. 406, 268 Ortega, г с. 4175 (1а). Padilla, 5. А. 881 (15). Palmer, Е. М. (1а); 132, 1470 (1a). Pennell, F. W. 4755 (12). [Vor. 15 ANNALS OF THE MISSOURI BOTANICAL GARDEN . Е. У. & Rusby, H. Н. 186 PA ud Н. 1177 (4); 183, 2908 (44); 11094 (5); 11, 1594, 2824, 3388, 3739, 8940, 4406, 4748, 5011, 6355, 7774, 10288, 10616 (10); 8617 (14); 1936 (15). Popenoe, W. E Pringle, C. G. 20 (1); 6161, 5363, 6847, 6872 um 8103 (7). Ригрив, С. А. 6230, 8771 (4а); 7740 (7). Кеко, В. Р. 4892 (12). René, А. 78 (9). Кепвоп, С. 106 (15). Ricard, В. Н. 13 (15). Т” А. & Гуоп, М. У., Л. — (10 Rose, 2. М. 1578, 1688, 2888, 8960 (1a); —, 1614 (9); —, 1711 (10). Rose, J. N. & Hough, W. 4777 (1a). Rose, J. N., Painter, J. H. & Rose, J. S. 9274 (4a). Rose, J. N. & Rose, G. 23612 (4a). Rose, J. N., Standley, P. C. & Russell, P. G. 13874 (1); 14064, 14544, 14675 (1a); 14484 (10). Rusby, H. H. 931, 1163 (4a). Samuels, J. A. —, 384, 386 (10). Schott, A. 430 (7). Seler, C. 3436 (1a). Shannon, W. C. 5042 (4a); 120, 152 (15) Smith, C. L. —, (3); 126, 1339, 1374 (4a). Smith, H. H. 1639 (10). Standley, P. C. 20686, 20713, 20948, 32798, 35459 (3); 21084 (4); 22372, 22539 (4a); 28840, 40994 (10); 19187, 21463, 21865, 29218, 23452, 2462 40220, 40994 (15). Stevens, F. L. 90 (10). Tejera, E. 14 (10 duz, A. 13900 (4a); 17663 (5); 13904 (15). Tonduz, A. & Rojas, A. 14? (7); 17653 (10); 36 (15 Tuerckheim, H. von. 981 (7). Valerio, J. 86 (15). Weigelt, C. — (10). Williams, К. S. 803 (10). 1928] WOODSON-—STUDIES IN APOCYNACEAE, II 373 INDEX TO SPECIES ew subgenera, sections, species, varieties, and combinations are printed in bold bon type; synonyms in italics; and previously published names in ordinary 1 Page Bignonia ЕО ИО 2100 857 Echites МапотаейЙота................. 361 Eustemmadenia............... 353 6 Galeottiag.................... 359 Malouetia guatemalensis................. 371 О О р И 371 $ оротайаё..................... 358 Ochrodaphne.................. 363 Odontostigma .................... 352 EOE T, T T 0а... 361 Stemmadenia................... 352 il en PEDE 360 DEA. ULL e 2 C 361 МапотаеЙоға................. 36 ОИ 12 25.2 4 оо ENS 858 beitia Ус 363 Donnell-Smithii............. 369 eubracteata................. 368 А... vou се nee 361 DEO NR НЕЕ. 356 grandiflora 364 ee 360 guatemalensis... Page ae aren сум та 361 macroplil ыы > n зА ЕЛА SER 362 РОО paren ^ ОИК 358 ek Пе и 357 obovata Var. ПОВ 358 23 Kose.” уа а А 354 СНО ПИ СД даа ын 354 “Palmeri Боне & Standl.". 354 рачсшогае 2... Sees 366 Pennellii. HIPS o ЕНЕ 307 pUbescena ..... нь 357 hinsoni а. то 369 IT преса пар оно 13 356 ООВ. а ое 358 tomentosa var. Palmeri 354 S И ITE T 4А 352 СЄаїеой атит.................- 861 Tabernaemontana EON О ИЕ 360 Donnell-Smithü............... 369 onnell-Smithit B. 00817221818 ............ 369 acsi у ee 364 [У ог. 15, 1928] 374 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 47 Fig. 1. Habit of Stemmadenia glabra. X М. Fig. 2. Habit of Stemmadenia Galeottiana. X М. 3. Detail of floral mechanism of Stemmadenia Galeottiana. 2. Fig. 4. Habit of Stemmadenia grandiflora. X 5. Fruit of Stemmadenia tomentosa var. Palmeri. х м. Ann. Мо. Вот. Garp., Vou. 15, 1928 PLATE 47 WOODSON—STUDIES IN APOCYNACEAE [V or. 15, 1028 376 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 48 Fig. 1. Stemmadenia sinaloana Woodson, from the type specimen, F. Н. Lamb No. 467, in the Gray Herbarium of Harvard University. Fig. 2. Stemmadenia Greenmanit Woodson, from the type specimen, Brenes 14875 in the Gray Herbarium of Harvard University. 48 PLATE ANN. Мо. Bor. Garb., Vor. 15, 1928 WOODSON—STUDIES IN APOCYNACEAE [Vor. 15, 1928] 378 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 49 Fig. 1. Stemmadenia pauciflora Woodson, from the type specimen, Pennell & Rusby No. 186, in the Herbarium of the New York Botanical Garden Fig. 2. Stemmadenia eubracteata Woodson, from the type specimen, Ноја & Lux No. 4538, in the Gray Herbarium of Harvard University. NI SWIGALS—NOSGOOA OVNADOdV ЧУН Ann. Мо. Вот. Слко., Vor. 15, 1928 PLATE 1 ) STUDIES IN THE APOCYNACEAE. Ш A MONOGRAPH OF THE GENUS AMSONIA ROBERT E. WOODSON, JR. Rufus J. Lackland Research Fellow in the Henry Shaw School of Botany of Washington University HISTORICAL Discussion When the second edition of Linnaeus’s ‘Species Plantarum’ appeared in 1762, one of the many additions to the species pre- sented in the first edition (1753) was Tabernaemontana Am- sonia, a plant the exact genus of which Linnaeus himself was not precisely sure, qualifying it to the genus T'abernaemontana with the remark “Affinis Camerariae et Tabernaemontanae.”’ The attitude with which Linnaeus treated his Tabernaemontana Amsonia is essayed by Sir J. E. Smith,? and throws much illu- mination upon problems concerning the genus which will receive subsequent treatment in this monograph: “‘Tabernaemontana The herbaceous plants, supposed by Linnaeus to belong to this genus, constitute, as we have already said, and as Lin- naeus himself originally thought, a very distinct one, of which we shall now treat by the name of Amsonia. We can give no positive account of the meaning or origin of this word except that its author, according to Miller,?! was Clayton. Linnaeus in his own copy of Gronovius’ Flora Virginica, ed. 1. p. 26, has written Amsonia as a generic name, to what Clayton took for a species of Neriwm, and has subjoined also in manuscript the characters of the follicles and seeds. This plant, in the second edition of the Species Plantarum, is the Т. Amsonia; and so it remained until Mr. Walter restored it to rank as a genus; but without throwing any light upon the name.” The name Amsonia has indeed been an enigma, and Вайп- ! L. Sp. РІ. ed. 2, 2: 301. 1762. 2 Sm. in Rees, Cycl. 35. 1819 3 Miller, Gard. Dict. ed. 5, 27: art. “Tabernaemontana.” 1807. Issued December 22, 1928. Ann. Mo. Вот. Garb., Vor. 15, 1928 (379) “ог. 15 380 ANNALS OF THE MISSOURI BOTANICAL GARDEN esque! even went so far as to change the name to “Ansonia,” referring to the passage above quoted from Smith, and naively remarking that he had been acquainted with several Ansons, but never an Amson, and so the name must be misspelled. With the fresh stimulus of Rafinesque’s contention a special search was made for the origin of the name Amsonia, and for a time it appeared that Rafinesque’s intuition had been well guided, for, although all of the published floras and manuals dealing with the genus spoke readily of “Ог. Amson, a colonial physician,” or “Charles Amson, a physician of South Caro- lina," no authentie trace of that gentleman could be found, either in published encyclopedias or standard reference works. Historical societies in Virginia and the Carolinas were invoked to no avail. An Amson, any Amson whatever, was not forth- coming. However, Ansons were frequent, including а certain Lord George Anson, a royal governor fond of explorative expeditions, from one of which he had discovered and brought to civiliza- tion a new esculent pea. Rafinesque was about to be vindi- cated, when a letter trom Clayton to John Bartram appeared which seems to solve the problem, although not completely. The letter, which was written from Gloucester County, Vir- ginia, Sept. 1, 1760, follows: “Dear Friend: “Т have sent you, enclosed, some seed of а new plant, which I presume is a stranger in your northern part of the world. Indeed it grows here only in the southern parts of the colony. I have it in my garden, but have quite forgotten whether I showed it to you, when I had the favor of your company. If I did, I believe I told you it was to be called Amsonia, after a doctor, here; but I think the name inscribed upon the inelosed more proper, as it answers to the particular form of its seed. *[ intend to send you some of the seed of our thorny Sensitive Plant by the first орнау that offers, after it is пре; * And remain, dear sir, your sincere friend “Апа most most humble servant, ыс “JOHN CLAYTON." ! Raf. New Fl. М. Am. 4: 58. 1838. 1928] WOODSON—STUDIES IN APOCYNACEAE. III 381 Allowing for orthographical errors, then, Amsonia was defi- nitely named for a certain Dr. Amson, a physician of Gloucester Co., Virginia; but regarding his complete name, or the positive form of spelling of his family name, doubt still remains. Lord. Anson, however, can undoubtedly be discarded as a possibility. The “пате inscribed upon the inclosed," which Clayton thought to be “more proper," was evidently Tabernaemontana Amsonia. Tabernaemontana Amsonia was immediately conspicuous among the other Tabernaemontanas both in habit and in hab- itat, since it was the only temperate herbaceous member of the genus, and іп 1788 attracted the attention of Thomas Walter,’ who described from it a new genus, naming the type species, in transposition of the Linnaean combination, Amsonia Tabernae- montana. At the same time, Walter? also described a new plant from the Carolinas which he assigned to the same new genus, calling it A. ciliata. As a result of his explorations in the southeastern United States, André Michaux? was able to expand the little genus with the addition, in 1803, of two new species, А. latifolia and A. angustifolia. The latter was a transfer to Amsonia of а species placed in the genus T'abernaemontana by Aiton* in 1789. Pursh,’ in 1814, also published a new species which he named А. salicifolia. Probably the most interesting addition which has ever been made to the species of Amsonia was in 1819, when Roemer and Schultes! transferred to that genus a plant which had been assigned to Табегпаетотата by Thunberg’ in 1784. The spe- cles, А. elliptica, was а native of Japan, and is yet the only known member of the genus not native to North America. Thunberg was slightly hesitant, as was Linnaeus, in committing his plant to the genus Tabernaemontana, but relied upon the precedence of the earlier author, remarking, much as did Lin- 1 Walt. Fl. Carol. 98. 1788. 2 Walt. l.c. 1788. 3 Michx. Fl. Bor. Am. 1: 121. 1803. «Ай. Hort. Kew. 1: 300. 1789. 5 Pursh, Fl. Am. Sept. ed. 1, 1: 184. 1814. * Roem. & Schult. Syst. Veg. 4:432. 1819. 1 Thunb. Fl. Jap. 111. 1784. [Уоь. 15 382 ANNALS OF THE MISSOURI BOTANICAL GARDEN naeus, "Valde affinis Amsoniae," a significant statement in the light of the later disposition of the species. The same year there appeared in the Rees 'Cyclopaedia' the description by Sir J. E. Smith! of a species of Amsonia which he called А. tristis. The plant was reported grown in an Eng- lish garden from seed collected in North America by Lyon, who contributed the plant from which Pursh published his A. salicifolia. Smith gave to his plant the common name “brown- ish-flowered Amsonia,” since, as he wrote, “Тһе flowers аге of а dingy brown hue, the segments of their limb одаи reflexed, at least in fading." The species has not been reported since its publication, and because of the rather suspicious- sounding description of the flowers, the name has generally been referred to as a synonym of А. Tabernaemontana Walt., or of A. salicifolia Pursh. Rafinesque's ‘New Flora of North America’ appeared in 1838, with a comparative sketch of the ‘‘Ansonias” then recognizable, and added one new species, A. tenuifolia,? the originality of which the author took unusual pains to indicate. In the ‘Prodromus’ of De Candolle the genus Amsonia re- ceived its first collective treatment. Besides recognizing the species which had previously been published with the exception of А. lenuifolia Raf., Alphonse De Candolle? described a new plant which he termed А. salicifolia Pursh var. ciliolata, from Alabama and Louisiana. The following year the genus gave evidence of the growing botanical knowledge of the southwestern United States by the publication by Torrey and Frémont! of A. tomentosa from “west of the Rocky Mountains.” Fourteen years later, as a result of the activities of the Mexican Bound- ary Survey, Torrey’ published a second species, A. longiflora, a very distinct plant of the region about El Paso, Texas. Some time later the genus came to the attention of Asa Gray during the course of the preparation of the ‘Synoptical Flora,’ 1 Sm. in Rees, Cycl. 35: end of art. “Tabernaemontana.” 1819. з Raf. New Fl. М. Am. 4: 58. 1838. * A. DC. in DC. Prodr. 8: 384. 1844. ‘Torr. & Frém. іп Frém. Rept. 1843-1844, 316. 1845. * Torr. in Rept. Mex. Bound. Surv. 21: 159. 1859 1928] WOODSON—STUDIES IN APOCYNACEAE. III 383 and in 1877! he published two new species from the Southwest, A. brevifolia and A. Palmeri. The following year when the ‘Synoptical Flora’? was issued, there appeared also а new variety of the genus which Gray termed A. angustifolia Michx. var. Texana. Besides being noteworthy for the contribution of a new variety, Gray’s treatment of Amsonia іп the ‘Synoptical Flora’ constitutes a scientific and comprehensive treatment of the group in its phylogenetic aspects. In 1894, in accordance with the Rochester Code of Nomen- clature, a double name was made by Britton? for the type spe- cies of the genus. This name, Amsonia Amsonia, is still current among some botanists. К. Schumann,‘ in Engler and Prantl’s ‘ Natiirlichen Pflanzen- familien,’ elaborated upon Gray’s treatment in the ‘Synoptical Flora’ and divided the genus into two sections which he called Euamsonia and Sphinctosiphon. In the first section, three spe- cles were recognized, namely, A. Tabernaemontana Walt., A. ciliata Walt., and A. elliptica (Thunb.) Roem. & Schult., and in the second section, four species, A. Palmeri Gray, A. longi- Лота Torr., A. brevifolia Gray, and A. tomentosa Torr. & Frém. In the twentieth century numerous additions have been made to the genus Amsonia. In 1900 A. A. Heller’ elevated Gray’s A. angustifolia Michx. var. Техапа to specifie rank. In Small’s ‘Flora of the Southeastern United States’ two new species are contained, A. ludoviciana Vail* and A. rigida Shuttleworth.’ Other new specific contributions have been A. latifolia M. Е: Jones,* 1908, А. Eastwoodiana Rydberg,’ А. arenaria Standley,” and A. hirtella Standley," in 1913. Jepson,” in 1925, reduced A. Lomentosa Torr. to a variety of A. brevifolia Gray. ! Gray, Proc. Am. Acad. 12: 64. 1877. ! Gray, Syn. Fl. М. Am. 2:: 81. 1878. Britton, Mem. Torr. Bot. Club 5: 262. * K. Sch. in Engl. & Prantl, Nat. Pi 4: 143. 1895. $ НеЏег, Мићепђегрла 1: 2. 1900. в Vail, in Small, Fl. Southeast. U. S. 935. 1903. 7 Shuttlew. in Small, 1. с. 1903. * Jones, Contr. West. Bot. 12: 50. 1908. • Rydb. Bull. Torr. Bot. Club 40: 465. 1913. 10 Standl. Pr а бос. Wash. 26: 117. 1913. u Standl. 1. c. 1 Jepson, Зэв И РІ. Cal. 768. 1925. (Мог. 15 384 ANNALS OF THE MISSOURI BOTANICAL GARDEN Evidently the first printed illustration of an Amsonia was one . presumably of A. Tabernaemontana in Plukenet, t. 115, fig. 8, 1769, where it appears as “Apocynum Virginianum Asclepia- disfolio erectum floribus pallide caeruleis radici crassa." Because of the peculiar distribution of the genus as well as because of its growing need for a taxonomical revision, it was thought appropriate that а rather broad study be made of the genus Amsonia. Such a study was begun at the Gray Her- barium of Harvard University under the oversight of Dr. B. L. Robinson and Prof. M. L. Fernald, and completed at the her- barium of the Missouri Botanical Garden under Dr. J. M Greenman. То Professors Robinson, Greenman, and Fernald, the author wishes to express his obligations most heartily for their kindly criticism and their ready suggestions. To Mr. T. H. Kearney the author is also indebted for much valuable aid with regard to the difficult species of the southwestern United States. Various herbaria have been visited, also, or specimens have been borrowed, and to the curators of each the author would express his gratitude. Gross MORPHOLOGY The genus Amsonia is one of the few members of the Аро- cynaceae which are temperate or subtemperate, and contains within its several species only erect perennial semi-woody herbs. Roots.—The root system of the group is characteristically fibrous. The crowns usually become woody with increasing age, and produce numerous clustered stems. Latex tubes occur in abundance, as in all the members of the family. Stems.—The stem system is typically that of an erect peren- nial and varies relatively little. A mature stem is usually di- vided into several branches. The species inhabiting the arid regions of the southwestern United States and northern Mexico frequently have stems which are branched to a much greater extent, and much lower upon the stem than the more temperate species of the states of the Southeast and Middle West. Leaves.—The leaves of the genus are alternate to subverti- cillate, and vary greatly in size and outline. Through the suc- cession of species, extremes are found in the leaves of А. Tab- ernaemontana, which are broadly ovate-elliptic, usually measur- 1928] | WOODSON—STUDIES IN APOCYNACBAE. ТП 385 ing 3-5 cm. long and 1.5-2.5 cm. broad, petiolate and opposite, to the subverticillate leaves of A. salpignantha, which are linear- lanceolate to linear-filiform, measuring 2-5 cm. long and .5-4 mm. broad, and decidedly sessile. Тһе leaves may also be gla- brous to glaucous, as they are in A. salicifolia, or densely to- mentose, as they occur in A. tomentosa. The leaves are always entire, and are never cordate. In only one species, A. Тафегпае- montana, are the bases of the leaf-blades other than acute when a petiole exists. Inflorescence.—The inflorescence is а thyrsoid or corymbose cyme. The amount and shape of the inflorescence, however, is varied. The largest inflorescence of the genus is found in A. Tabernaemoniana var. Gattingeri, which frequently contains over fifty blossoms, and the smallest іп A. Palmeri, which usually has only five or six. The inflorescence may have very incon- spicuous bracteoles, as in the subgenus Еиатзота, or quite conspicuous bracteoles, giving the whole inflorescence a chaffy appearance, as in the subgenera Sphinctostphon and Articularia. The inflorescence may also be surrounded by the foliage, as in A. arenaria, or held high above the foliage by a long, nearly leafless stalk, as in A. ciliata var. tenuifolia. Pedicels may be relatively long, as in A. Tabernaemontana var. salicifolia, or frequently lacking altogether, as in A. longiflora. Calyx.—The variation in the calyx is marked. In A. Табегпае- montana var. salicifolia the calyx is 1 mm. long or less in en- tirety, the lobes being minutely triangular-ovate. In A. tomen- tosa the calyx is as shallow as in the former species, but the lobes are fully 3-5 mm. long and are subulate-aristate. The calyx may be glabrous or pubescent, occasionally becoming sparsely hirsute. Corolla.—The corolla is regularly five-lobed. Тһе tube dilates upward, and may be unconstricted, as in the subgenus Еиат- sonia, or markedly constricted at the mouth, as in the subgenera Sphinctosiphon and Articularia. Variation in the length of the tube is great, ranging from 6-8 mm. in A. ciliata var. tenuifolia and allied species, to 3-3.5 cm. in A. longiflora and A. salpignantha, which have the most conspicuous flowers of the genus. The color variesfrom a clear cerulean blue, tinged to tawny-white in the tube, [Vor. 15 386 ANNALS OF THE MISSOURI BOTANICAL GARDEN in most of the eastern species, to white or a faint livid greenish blue in some of the western species. The tube is always villous within, and may be pubescent or glabrous without. The lobes of the cor- olla are spreading, and may be ovate to narrowly lanceolate in out- line. The length of the lobes varies from one-half the length of the tube to an equal length, save in the large-flowered species of the section longiflorae of the subgenus Sphinctosiphon, where the ratio of the length of the tube to that of the lobes may be from about 3:1 in A. longiflora to 5:1 in A. salpignantha. Stamens.—The stamens number five, and are adnate to the corolla-tube well above the middle. The anthers are ovate- lanceolate, acute above, obtuse below, unappendaged, and fertile throughout their entire length. The stamens are free from the stigmatic-cap. Pistil.—The two carpels of the gynoecium, which are unilocu- late and contain many two-seriate anatropous ovules, are united by & common filiform style, which is about the length of the corolla-tube, to a position immediately below the stamens, where it is surmounted by а stigmatic-cap bearing the stigma. The stigmatic-cap is constructed in three elements, the lower of which is a reflexed membranaceous appendage, originating from the summit of the stylar shaft, the central, a tangled mass of short papillae, and the upper, the stigma itself, which may be depressed-capitate or truncate, as in the subgenus Huamsonia, or apiculate by two distinct obtuse lobes, as in the subgenera Sphinctosiphon and Articularia. Fruit.—The fruit of Атзотла is а pair of follicles which are cylindrical and acuminate, and may be slender and continuous, as in the subgenera Еиатзотла and Sphinctosiphon. or torose and definitely articulated into thickish constricted segments, as in the subgenus Атисшата. In either case the seeds are one- seriate, cylindrical, and unappendaged, but in Articularia the endosperm is conspicuously thicker and more corky than in the endosperm of the other subgenera. SYSTEMATIC POSITION The genus Amsonia is placed in the tribe Plumeroideae of the Apocynaceae because of its free unappendaged stamens. 1928] WOODSON-—STUDIES IN APOCYNACEAE. Ш 387 The characters of an ovary containing six to many ovules, an eglandular calyx, coriaceous fruit, a hypercraterform corolla, and . included stamens moreover place the genus in the subtribe Euplumeroideae. The closest related genus to Amsonia appears to be for various reasons Haplophyton. The two genera are found in common territory from southern California to southwestern Texas. Мог- phologically the greatest dissimilarity lies in the seeds which are appendaged in Haplophyton. The leaves, which serve to aid the differentiation of the two genera in the ‘Synoptical Flora," are not as widely separated as is ordinarily to be sup- posed, since they are not absolutely opposite in Haplophyton and alternate in Amsonia, but are more nearly approximate in the former and frequently subverticillate in the latter. The stamens are nearly alike in both genera, but are somewhat larger in Haplophyton. The character of the stigmatic head in that genus is also much like that of the stigmatic-head in the subgenus Еиатзота of Amsonia, although more elongate, but lacks a membranaceous reflexed appendage. However, a distinct swollen region occurs upon the stylar shaft of Haplo- phyton just below the papillose cap, which might be regarded as a primitive stage in the development of the more elaborate appendage of Amsonia. Rhazya is also a genus closely related to Amsonia, but possesses a disc and a jointed clavuncle among other dissimilarities. RELATIONSHIP AND DISTRIBUTION OF THE SUBGENERA The genus Amsonia, although relatively a small group, is readily separable into three subgenera, which, while interlocking closely, are distinct and well differentiated entities in the whole. The series of subgenera range in geographical and evolutionary succession from east to west and south in North America, upon which continent the bulk of the species occur, only one species being found in eastern Asia. The subgenus Euamsonia is the largest of the divisions in number of species and varieties and the most widely spread, embracing five species and four varieties in the southeastern United States, and one species in Japan. The second largest 1 Gray, Syn. Fl. М. Am, 2:: 81. 1878. (Vor. 15 388 ANNALS OF THE MISSOURI BOTANICAL GARDEN subgenus with regard to number of species and extent of dis- tribution is Sphinctosiphon, which occurs with eight species in the central-southwestern United States and adjacent Mexico, having for its center of distribution southern New Mexico and northern Chihuahua. Articularia is the smallest of the sub- genera, and contains four species limited to southern California, southern Nevada, southwestern Utah, and western Arizona; while one species, A. arenaria, is isolated from the general dis- tribution of the subgenus to which it belongs, in extreme south- western New Mexico and adjoining Chihuahua. The situation of Huamsonia in having species of the south- eastern United States and Japan is by this time of more or less frequent knowledge, and no speculations will be devoted to it, since similar instances have been reported." ? "€, The occur- rence of the three subgenera in the southern United States and northern Mexico is, however, of general interest. A study of the genus in North America suggests forcibly that it is a genus of mesophytic origin which exhibits an increasing adaptation to an arid habitat. Еичатзотла is the one subgenus of a mesophytic habit, and since it is represented by the relict species in Japan to which reference has already been made, it is taken as the most primitive. The subgenera Sphincto- siphon and Articularia are plants of distinctly arid habitat, and the morphological differentiation which those groups ex- hibit are interpreted as divergences from the primitive condition represented by Huamsonia. The genus Amsonia is relatively advanced among the Plumer- oideae because of its highly differentiated stigmatic-cap, among other characters, and it is upon the basis of further differentia- tion in that respect that the first subgeneric division is made. In the subgenus Huamsonia the stigma proper is depressed- capitate or truncate, and appears merely as the freer summit of the papillose central region of the stigmatic-cap to which reference was made in detail in the previous section concerning Gross Morphology. The mouth of the corolla-tube, moreover, is relatively open, continuing the dilation of the tube. In the 1 Gray, А. Mem. Am. Acad. М. 5. 6: 377-449. 1859, з Fernald, М. L. Quart. Rev. Biol. 1: 227. 1926. 1928] WOODSON—STUDIES IN APOCYNACEAE. III 389 subgenus Sphinctosiphon an evolutionary advance is detected in the elevation of the stigma to the position of two distinct apiculate lobes, and the constriction of the mouth of the corolla- tube. Such differentiations are obviously of use to the plant for insect pollination in an arid region. The subgenus Articularia demonstrates a further advance in the articulation of the follicles, which are quite slender and continuous іп Zuamsonia and Sphinctosiphon, into thickish con- stricted segments in much the same manner as the legumes of certain desert Leguminosae, beside having the apiculate char- acters of the stigma. The seeds of the follicles of Articularia, moreover, are larger and ovoid, and the endosperm is thick- ened, but of a light and corky texture, an evident construction to facilitate easy dissemination in an arid habitat. The seeds of Euamsonia and Sphinctosiphon, on the other hand, are roughly cylindrical with a relatively thin, hard endosperm. The subgenera of Amsonia are remarkable for their inter- rupted distribution, a factor which lends even sharper distinc- tion to the morphological differences which they display, and suggests certain hypotheses for their origin. Еиатзота, with the greatest number of species and varieties, has been found naturally in all of the southeastern United States, with a gener- ally characteristic habitat of moist woods, ravines, or stream- sides, save in its extreme western limits, where A. ciliata var. texana is found on rocky hillsides and prairies. Sphinclosiphon, with the next largest number of species, is confined to southwestern Colorado, southeastern Utah, New Mexico, and adjacent portions of Arizona, Chihuahua, and Texas. Thus Sphinctosiphon and Euamsonia are entirely sepa- rate in range, except in south-central Texas, which contains two species of Euamsonia and a limited colony of А. salpig- naniha of Sphinctosiphon. Besides the anomalous occurrence of A. salpignantha within the southwestern limits of Huamsonia the nearest that the subgenera approach each other is evi- dently in western Texas, Euamsonia being found in the Wichita Mountains of north-central Texas, and Sphinctosiphon in the Guadaloupe Mountains about two hundred miles to the south- west. The species of Sphinctosiphon, although in an arid region, [Vor. 15 390 ANNALS OF THE MISSOURI BOTANICAL GARDEN partake of the nature of Euamsonia in frequenting the borders of ponds, streams, and branches. Articularia, with the fewest number of species, is confined to southern California, southern Nevada, southwestern Utah, and northwestern Arizona, save for the species А. атепата, which has а distribution analogous to the anomalous distribution of А. salpignantha of Sphinctosiphon, occurring separate from the other species with which it has its affinities, in extreme south- western New Mexico (Grant County), and adjacent Chihuahua, within the distributional area of Sphinctosiphon. Тһе species of Articularia demonstrate the most extreme endurance for aridity, being found most frequently in the sand of the open desert, whence, it is rather safely supposed, occurs the striking mor- phological adaptations which they exhibit. Thus Sphinctosiphon and Articularia possess rather distinct areas of distribution, save for A. arenaria of Articularia which occurs fully three hundred miles, to present knowledge, from the known range of its kindred species, а perplexing situation. It is also possible that А. Hastwoodiana and A. Jonesii, species of Articularia and Sphinctosiphon respectively, meet in southern Utah and northern Arizona. At any rate, А. Kearneyana, oc- curring in regions midway between the territories of Sphincto- siphon and Articularia, for reasons which will be advanced later, appears in all probability an hybrid between A. Palmeri of the former subgenus and A. brevifolia of the latter. Thus it is seen that the subgenera of Amsonia occupy essentially distinct and isolated ranges, Articularia and Sphinctosiphon, the most nearly related of the groups morphologically and ecologically, being also the most neighborly distributed, and both distinctly removed from Euamsonia, the supposed primitive subgenus, morpho logically and geographically. If we are to believe that by the Cretaceous the modern angio- spermous type of vegetation had become fully established throughout the world,' we may assume that the genus Amsonia was by that time in а flourishing condition with a wide distri- bution over the southern half of what is now the United States 1 Grabau, А. УУ. Textbook of geology 2: 687. 1922. 1928] 4 ТЕПП Pree ЛИК М ХР TN Кем à НА pu SN әй” АХ SN FU У 4 fi E. NORTH AMERICA (STATE BOUNDARIES) SCALE OF MILES DUE CUm лан ла” 120 по Fig. 1. Land mass of North America in Comanchean time. and adjacent Mexico. The position of the land masses in !Stopes, M. C. Ancient plants, p. 85. 1910. Dr. Stopes wrote, in support of such an assumption: ‘‘Specimens of Cretaceous plants from various parts of the world seem to indicate that there was a striking uniformity in the flora of that period all over the globe." [У ол, 15 392 ANNALS OF THE MISSOURI BOTANICAL GARDEN SN N N RA ~ 5-9 NORTH AMERICA (STATE BOUNDARIES) SCALE OF MILES Пипи жән Yeo Fig. 2. Land mass of North America in late Comanchean time. Comanchean, or lower Cretaceous, time would lend support to the speculation that the genus was allowed at that time practically an uninterrupted range (fig. 1), and because of that reason was very likely of a more or less uniform character. The 1928] WOODSON—STUDIES ТМ АРОСУМАСЕАК. III 393 fact that Ше genus is now found as a relict in Japan is reason enough for assuming a wide range for its species. | During the late Comanchean time, however, the continuous range supposed for the genus was broken by the inundations of the Colorado Trough (fig. 2), which occurred over nearly the whole of northeastern Mexico, Texas, and parts of Oklahoma, Colorado, Kansas, and New Mexico. This invading sea could scarcely be regarded as less than a most effective opportunity for generic variation through isolation, especially since the vege- tation in the isolated mass was by that time bearing evidence of an adaptation to aridity.! The Cretaceous time, proper, is well known for the extensive inundations which then occurred widely in North America, and the break in the hypothetical distribution of Атзота was heightened by an increase of the seas of the Colorado Trough, which cut completely through the continent from what is now the coast of the Territory of Mackenzie to the Gulf of Mexico. Troughs of the western coast also caused intrusions during the early periods of the Cenozoic, reaching a climax during Mio- cene time, when large tracts of southern California and adja- cent Lower California and Arizona were separated as islands (fig. 3). By the Pliocene time, North America had largely assumed the shape with which we are now familiar. With isolated land masses corresponding roughly to the lo- calities of probable origin of the subgenera Sphinctosiphon and Articularia, a fair degree of credence might be allowed the assumption of their differentiation upon those lands, the first instance of isolation, the intrusion of the Colorado Trough, possibly giving rise to the development of the type of Sphincto- siphon from the primitive condition represented at present by Euamsonia, and the second, the production of islands by the inundations of the west coast troughs during the Miocene, pro- viding an opportunity for the divergence of the type of Articu- laria from the group now represented by Sphinctosiphon. It is 1 Schuchert, C. Outlines of historical geology. 1924. “Іп general we may say that after the early upper Cretaceous time . . . the climate the world over was . . . warm temperate in character [p. 612.] With the Miocene, however . more or less of desert climates developed in the Cordilleran areas of North Ханиа and have prevailed there ever since (р. 626.” [Vor. 15 394 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Ч E Т pue mir if МАП % ех EUN 5 ТІ РЯ Г ха с 77 ша ча Мае А PADS Т NORTH AMERICA (STATE BOUNDARIES) SCALE OF NILES 120 по Fig. 3. Гапа mass of North America in upper Cretaceous time. believed that the discontinuous areas of the subgenera support the hypothesis offered for their origin. The fact that the dis- tribution of the three subgenera still bears evidence of disrup- tion thousands of years after the Mesozoic and Cenozoic inun- dations appears quite striking, and bears additional evidence of 1928] WOODSON—STUDIES IN APOCYNACEAE. III 395 the old аве! of the genus in its inability to reunite its former distribution. In this respect, the genus Amsonia offers an in- teresting parallel to the endemics of the unglaciated regions of boreal America, whose ranges were broken by the Pleistocene glacial phenomena: ‘‘These older species in North America have long since passed their period of aggressiveness. ‘Left undis- turbed they persist in their old habitats, but they fail to move into new and immediately neighboring territory.’ ”? ABBREVIATIONS Abbreviations indicating the herbaria where specimens cited in this monograph are deposited are as follows: Baker = C. F. Baker Herbarium of Pomona College. Е = Field Museum of Natural History Herbarium. G = Gray Herbarium of Harvard University. MBG = Missouri Botanical Garden Herbarium. МЕ = New England Botanical Club Herbarium. NY = New York Botanical Garden Herbarium. г = Pomona College Herbarium. ANSP = Academy of Natural Sciences of Philadelphia Her- barium. РВС = Philadelphia Botanical Club Herbarium. TAXONOMY Amsonia Walt. Fl. Carol. 98. 1788; Michx. Fl. Bor. Am. 1: 121. 1803; Pursh, Fl. Am. Sept. ed. 1, 1: 184. 1814; Roem. & Schult. Syst. Veg. 4: 432. 1819; Smith in Rees, Cycl. 35: end of art. “Tabernaemontana.” 1819; Elliott, Sketch Bot. 5. С. & Ga. 316. 1821; Endl. Gen. Pl. 582. 1838; A. DC. in DC. Ргодг. 8: 384. 1844; Pfeiffer, Nom. Bot. 11: 156. 1873; Benth. & Hook. Gen. Pl. 2: 703. 1876; Gray, Syn. Fl. М. Am. 2': 81. 1878; Durand, Index Gen. Phan. 262. 1888; Baill. Hist. Pl. 10: 180. 1891; Coulter, Contr. U. S. Nat. Herb. 2: 262. 1892; Coville, Contr. U. S. Nat. Herb. 4: 142. 1893; ! The presence of Amsonia іп the Cretaceous period would give to the genus ап age, based upon the most capable of present calculations (Schuchert, С. 1. с. 485. 1924), of at least 45,000,000 years. з Fernald, М. Г. Persistence of plants іп unglaciated areas of boreal America. Mem. Am. Acad. 15: 244. 1925; Antiquity of vascular plants. Quart. Rev. Biol. 1:227. 1926 [Vor. 15 396 ANNALS OF THE MISSOURI BOTANICAL GARDEN К. Schumann in Engl. & Prantl, Nat. Pflanzenfam. 4?: 143. 1895; Mohr, Contr. U. S. Nat. Herb. 6: 674. 1901; Small, Fl. Southeast. U. 5. 934. 1903; Dalla Torre & Harms, Gen. Siph. 406. 1904; Harper, Ann. N. Y. Acad. Sci. 17': 175. 1906; Robinson & Fernald in Gray, New Man. ed. 7, 661. 1908; Nelson in Coulter & Nelson, New Man. Rocky Mt. Bot. 385. 1909; Matsumura, Index Pl. Jap. 2: 505. 1912; Wooton & Standley, Contr. U. S. Nat. Herb. 19: 504. 1915; Rydb. FI. Rocky Mts. 668. 1917; Davidson & Moxley, Fl. South. Cal. 278. 1923; Tidestrom, Contr. U. S. Nat. Herb. 25: 418. 1925; Jepson, Man. Fl. Pl. Cal. 768. 1925. Ansonia Raf. New Fl. N. Am. 4: 58. 1838. Lactescent herbaceous caulescent perennials, glabrous or pu- bescent. Leaves alternate or subverticilate, sessile or petio- late, membranaceous or somewhat thickened and fleshy, entire. Inflorescence a terminal thyrsoid or corymbose cyme. Calyx five-parted, the lobes acuminate or subulate. Corolla salver- form, villous within, tube cylindrical, dilating, open or con- stricted; lobes ovate to lanceolate, spreading or nearly erect. Stamens five, adnate to the corolla-tube above the middle, included; anthers ovate to ovate-lanceolate, obtuse, unappen- daged. Disk wanting. Carpels two, united by the filiform style surmounted by a truncate stigmatic-cap. Stigma de- pressed-capitate, or apiculate by two distinct lobes, surrounded by a spherical papillose mass, and appendaged by a reflexed membrane. Ovules in each carpel many, two-seriate. Follicles two, cylindrical, continuous or articulated. Seeds many, one- seriate, cylindrical, unappendaged. Embryo straight. Type species: А. T'abernaemontana Walt. Fl. Carol. 98. 1788, SYNOPSIS OF THE SUBGENERA AND SECTIONS KEY TO THE SUBGENERA A. Stigma depressed-capitate or truncate............... Subgenus I. Evamsonia B. Stigma apiculate by two distinct lobes. a. Follicles continuous, not articulated.......... Subgenus II. БРНТСТОВТРНОМ b. Follicles torose, articulated into thickish constricted segments....... ИТР ubgenus Ш. ARTICULARIA Бовавков I. Evamsonia (К. Schumann) Woodson Subgenus I. Еслмвоміл (К. Schumann) Woodson, п. comb. $Еиатзотла К. Schumann in Engl. & Prantl, Nat. Pflanzen- 1928] WOODSON—STUDIES IN АРОСУМАСЕАЕ. Ш 397 fam. 4°: 143. 1895; Dalla Torre & Harms, Сеп. Siph. 406. 1904. h Bracteoles inconspicuous; orifice of the corolla-tube not con- stricted in anthesis; stigma depressed-capitate or truncate; fol- licles slender and continuous, not articulate, fibrous, not horny in texture; seeds irregularly oblong in outline, truncate at either end, variously pitted and wrinkled; plants of the southeastern United States and Japan. Spp. 1-5. Ккү то THE SPECIES AND VARIETIES a. Corolla glabrous withou b. Leaf-blades 4 ‘distinctly petiolate throughout.......... 1. A. rigida bb. БЭ Креда 7 to linear, sessile or subsessile above. Corolla-tube 6-8 m d. Stem-leaves Сы Би long as broad; infl held above the tolase. очнь ГА 4. ciliata dd. Stem-leaves 15-30 times as long as | реж. high above the foliage.......... 2a. A. ciliata var. нэ ЗА! се. Corolla-tube 9-12 mm. long. d. са about half as long аз Ше tube; pedicels 3-5 m. long; species of North America. .2b. А. yrs фа var. terana dd. Согой 0568 about equalling Ше tube; pedicels 5-10 ong; према Of Да, се 0c ос 8. Ч чина aa. Corolla pubescent without b. Follicles glabrous с. Leaf-blades ovate to ee the bases of the low obtuse to broadly асше.................. 4. A. Ta ‚роде ВИ се. Leaf-blades ucl: 6 linear-lanceolate, the bases of the lower acute to acuminate. d. Inflorescence loose, few-flowered; foliage glabrous, glaucous спра... es сова 4a. А. abernaemontana var. salicifoliá dd. тар dense, many-flowered; foliage pubescent, gla- ee E 4b. A. Tabernaemontana var. Gattingeri bb. Follicles Е at least upon Ше upper portion....... 6. А. ludoviciana 1. Amsonia rigida Shuttlew. in Small, Fl. Southeast. U. S. 935. 1903; Harper, Ann. N. Y. Acad. Sci. 171: 175. 1906. Herbaceous perennial from a thickened somewhat woody root; stems 8-15 dm. tall, regularly branched above, glabrous; leaves alternate, numerous, the blades almost exactly elliptic, isophyl- lous, 7. e., the lower and the upper leaves of nearly like outline, green "A glaucous or glaucescent beneath, 2.5-6 cm. long, .5-1.5 em. broad, distinctly petiolate throughout; flowers rela- [Vor. 15 398 ANNALS OF THE MISSOURI BOTANICAL GARDEN tively numerous in fairly loose cymes; pedicels 5 mm. long or slightly less; calyx 1-1.5 mm. long, glabrous, the lobes triangu- lar-ovate; corolla salverform, the tube 6-8 mm. long, gradually dilating upwards, glabrous without, the lobes lanceolate, 7—10 mm. long, widely spreading; stigmatic-cap about as tall as broad, stigma depressed-capitate; follicles slender, continuous, gradu- ally attenuate, 7-11 cm. long, sessile, glabrous, 7-10-веедед; seeds 5-11 mm. long, oblong in outline, truncate at either end, variously wrinkled and pitted, dark brown. Distribution: swampy or moist pine forests, northern Florida and southern Georgia. Specimens examined: Скокоста: Alapaha, swampy pine woods, June 25, 1901, Cur- tiss 6820 (G, MBG, NY, US); Sumter Co., moist pine barrens, Aug. 21, 1900, Harper 448 (G, MBG, NY, US, F); same locality, Sept. 6, 1900, Harper 606 (NY, US); same locality, Aug. 21, 1900, Harper 440 (NY, US). ЕтовтрА: Chattahoochee, Мау, 1882, Curtiss (С); data lack- ing, Chapman (G, ANSP, MBG); St. Marks, June, 1843, Rugel (MBG); Chattahoochee, 1891, Chapman (MBG). 2. Amsonia ciliata Walt. Е. Carol. 98. 1788; А. DC. in DC. Prodr. 8: 385. 1844; Wood, Classbook Bot. 589. 1860; Chapm. Fl. South. U. S. 343. 1897; Mohr, Contr. U. S. Nat. Herb. 6: 674. 1901; Small, Fl. Southeast. U. S. 935. 1903; Harper, Апп. Х. Y. Acad. Sci. 171: 175. 1906. РІ. 51, figs. 7-8. Tabernaemontana angustifolia Ait. Hort. Kew. 1: 300. 1789; Willd. Sp. Pl. 12: 1247. 1798. Атзотла angustifolia (Ait.) Michx. Fl. Bor. Am. 1: 121. 1803; Pursh, Fl. Am. Sept. ed. 1, 1: 184. 1814; Roem. & Schult. Syst. Veg. 4: 432. 1819; Ell. Sketch Bot. S. C. & Ga. 317. 1821; Darby, Bot. South. States, 434. 1860; Gray, Syn. Fl. N. Am. 21:81. 1878. Ansonia ciliata (Walt.) Raf. New Fl. N. Am. 4: 58. 1838. Ansonia angustifolia (Ait.) Raf. l. c. 1838. Herbaceous perennial from a thickened woody root; stems 7-15 dm. tall, clustered from the base, erect or slightly ascend- ing, sparsely branched above, the branches ascending, pubes- 1928] WOODSON—STUDIES IN АРОСУМАСЕАЕ. ПТ 399 cent, glabrous ог glabrate in age; leaves numerous, crowded, subverticillate above, slightly heterophyllous, 1. е., the lower leaves broader and of a slightly different outline than the upper, linear-lanceolate, or the lower oblong-lanceolate, pubescent, or glabrate in age; inflorescence dense, barely held above the foli- age; pedicels 3-5 mm. long, sparsely pubescent; calyx 1-1.5 mm. long, glabrous, or with a few scattered hairs, the lobes triangular-ovate; corolla salverform, the tube 6-8 mm. long, glabrous without, the lobes 7-8 mm. long, oblong-lanceolate, erect or spreading; stigmatic-cap slightly broader than tall, stigma depressed-capitate or truncate; follicles slender, con- tinuous, 9-11 cm. long, gradually attenuate, sessile, glabrous, 7-11-веедед; seeds 5-11 mm. long, oblong in outline, truncate at either end, variously pitted or wrinkled, dark brown. Distribution: pine forests, occasionally entering fields; North Carolina, South Carolina, southern Georgia, northern Florida, southern Alabama, and northeastern Texas. Specimens examined: Ковтн Савошма: data lacking, Curtis (С). Ѕоотн CAROLINA: Aiken, April, 1882, Velden (MBG); Aiken, sand hills near Graniteville, May 7, 1899, Eggert (MBG); Aiken, May, 1869, Canby 68 (MBG, G, NY); Columbia, woods, May 9, 1899, Sargent (G); data lacking, Ravenel (G, NY, US); Colum- bia, dry sandy pine woods, Мау, 1890, Taylor (Е). Сковста: Richmond Co., slopes of sand hills about 8 miles west of Augusta, June 10, 1902, Harper 1319 (US, NY, F); Augusta, date lacking, Olney & Metcalfe 76 (G); data lacking, Wilkins (G). Fiorina: Tallahassee, date lacking, Berg (NY); Aspalaga, dry pine woods, April, year lacking, Curtiss 2269 (G, MBG, ANSP, S, F); River Junction, fields and open woods, April 22 and May 16, 1898, Curtiss 6376 (а, NY, US, MBG); data lacking, Chapman (G); Aspalaga, May, 1898, Chapman (MBG); Coffee Co., rocky open ground, flood plains of Pea River, May 15, 1925, E. J. Palmer 27233 (MBG); Chehaw, June 24, 1915, Drushel 4572 (MBG). ALABAMA: data lacking, Durand (ANSP). Texas: San Marcos, June 6, 1897, Stanfield (NY); Mid- [Vor. 15 400 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN lothian, April 30, 1895, Plank (NY); Turtle Creek, Kerr Co., date lacking, Bray 239 (US); Orange, April 17, 1899, Bray 60 (US). Although recognizing that A. ciliata Walt. antedates A. an- gustifolia (Ait.) Michx., Gray placed Walter’s species in sy- nonymy with the latter species, remarking that ciliata was an inappropriate name. The specimens with Gray’s labels in the Gray Herbarium truly are glabrate or glabrous, being overly matured specimens, hence Dr. Gray’s impression. In any event, Walter’s name can scarcely be discarded. 2a. Var. tenuifolia (Raf.) Woodson, n. comb. Ansonia tenuifolia Raf. New Fl. N. Am. 4: 58. 1838. Amsonia salicifolia Pursh var. ciliolata A. DC. in DC. Prodr. 8:384. 1844. Amsonia ciliata Walt. var. filifolia Wood, Classbook Bot. 589. 1860. Amsonia tenuifolia (Raf. Harper, Ann. N. Y. Acad. Sci. 17:: 175. 1906 Herbaceous perennial from a fibrous root; stems 3-10 dm. tall, single or sparingly clustered from the base, erect or slightly ascending, sparingly branched above, the branches ascending, pubescent or glabrate in age; leaves numerous, crowded, sub- verticillate, scarcely heterophyllous, 2. е., the lower leaves barely broader and of about the same outline as the upper, linear- lanceolate to filiform, pubescent or glabrate; inflorescence dense, held high above the foliage by a slender, usually leafless stalk; pedicels 3-5 mm. long, barely strigose or glabrous; calyx 1-2 mm. long, glabrous, or with a few short hairs, the lobes triangu- lar-attenuate; corolla salverform, the tube 6-8 mm. long, gla- brous or slightly canescent without, the lobes 4—6 mm. long, ovate to oblong-lanceolate, erect or spreading; stigmatic-cap about as tall as broad, stigma depressed-capitate or truncate; follicles slender, continuous, 8-14 cm. long; seeds 7-12 mm. long, oblong in outline, truncate at either end, variously pitted or wrinkled, brown. Distribution: sand-hills and barrens, also rocky margins of streams; North Carolina, South Carolina, Georgia, Florida, Ala- 1928] WOODSON—STUDIES IN APOCYNACEAE. III 401 bama, southern Arkansas, Missouri, Texas, and central Mexico. Specimens examined: UNITED STATES: Ковтн CAROLINA: data lacking, Curtis (С); White Hall, May 13, 1896, Biltmore 1400 (US). бостн CAROLINA: Aiken, May 21, 1899, Eggert (MBG). Скокста: Altamaha, sand-hills, date lacking, Chapman (С); Augusta, sand-hills, June 10, 1902, Harper 1319 (G, MBG, NY); Bainbridge, low woods bordering Flint River, July 13, 1899, Curtiss 6476 (G, MBG, NY, US); Bulloch Co., sand-hills along Big Lott’s Creek, June 17, 1901, Harper 915 (G, MBG, NY, US, F); Camilla, Mitchell Co., dry sand barrens, Aug. 7, Harper 1166 (G, NY, US); Dublin, Laurens Co., sand-hills of Oconee River, April 20, 1904, Harper 2188 (G, MBG, NY, US, F); Jasper City, 1846-48, Porter (G); Dooly Co., dry pine barrens near Gum Creek, Sept. 3, 1900, Harper 577 (US); Burke Co., Aug. 15, 1897, Hopkins 83 (NY); Thomson, McDuffie Co., sand-hills, Sept. 9, year lacking Bartlett 1493 (P); Vidalia, April, 1914, Huger (MBG); Macon, date lacking, Green (ANSP). ЕтовтрА: Bellair, Sept. 3, 1895, Nash 2546 (С, MBG, US, F); Clarcona, Orange Co., date lacking, Meislahn 210 (US); Gotha, March 28, 1919, Nehrling 12 (US); pine woods west of Jacksonville, April, 1848, Rugel 21 (US, MBG, F, NY); data lacking, 1873, Fell (ANSP); Cocoanut Grove, 1899, Rodman (С); Sumter Co., grassy pine-barrens, March 11, 1883, Donnell- Smith (С); “East Florida," date lacking, Buckley (С); “ Middle Florida," date lacking, Eaton (С); data lacking, Buckley (С); Alachua Co., June-July, 1898, Hitchcock (MBG); Lake Brant- ley, Aug. 1, 1895, Williamson (ANSP). ALABAMA: data lacking, Buckley (MBG); Bon Secour (near Mobile), June 29, 1893, Mohr (US). Missourt: Ozark Co., rocky open ground, bald knobs, near Tecumseh, Oct. 9, 1927, Е. J. Palmer 33081 (MBG); summit of bald knob across river from Tecumseh, Ozark Co., Nov. 11, 1928, Anderson & Woodson 4000 (MBG). ARKANSAS: Logan Co., rocky margins of small streams, Oct. 18, 1923, Е. J. Palmer 24203 (G); Hot Springs, Aug. 5, 1879, Letterman (MBG); Arkadelphia, May 10, 1884, Letterman (MBG). (Vor. 15 402 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Texas: data lacking, Wright (С); Medina Lake, Bandera Co., limestone ledges, creek banks, June 14, 1917, Е. J. Palmer 12262 (MBG); Johnson Co., rocky prairies, April, 1882, Re- verchon 84 (MBG). Mexico: Micnoacan: Morelia, June, 1901, Arséne (Е). 2b. Var. texana (Gray) Coulter, Contr. U. 8. Nat. Herb. 2: 262. 1892. Amsonia angustifolia Michx. var. Texana Gray, Syn. Fl. М. Am. 21: 81. 1878. Amsonia texana (Gray) Heller, Мынаа 1: 2. 1900; Small, Fl. Southeast. U. S. 935. 1903; Rydb. Fl. Colo. 269. 1906; Nelson in Coulter & Nelson, New Man. Rocky Mt. Bot. 385. 1909; Clem. & Clem. Rocky Mt. Fl. 100. 1914. Herbaceous perennial from a slightly woody root; stems 2-5 dm. tall, usually clustered from the base, erect or slightly as- cending or spreading, occasionally pubescent when young, mostly glabrous; leaves alternate, numerous, quite heterophyllous, 1. е., the lower leaves broader and of a different outline than the upper, ovate to oblong-lanceolate below, lanceolate to linear- lanceolate above, occasionally with short scattered hairs; in- Яогевсепсе compact, barely held above the foliage; pedicels 3—5 mm. long; сајух 1.5-2.5 mm. long, glabrous or glabrate, the lobes triangular-lanceolate to subulate; corolla salverform, the tube 9-11 mm. long, glabrous without, the lobes 4-6 mm. long, ovate to ovate-lanceolate, spreading; stigmatic-cap broader than tall, stigma truncate; follicles slender, continuous, 6-10 cm. long, rather abruptly acuminate, sessile, glabrous. 5-15-seeded; seeds 5-11 mm. long, oblong in outline, truncate at the ends, variously pitted or wrinkled, brown. Distribution: dry and rocky hillsides and prairies; Oklahoma, and Texas. Specimens examined: OKLAHOMA: Crusher Spur, Murray Co. , rocky mountain-side, April 12, 1913, Stevens 29 (G, MBG, US); Fort Sill, May 20, 1892, Sydone (NY); Tishomingo, on hillsides, common, April 8, 1916, Houghton 3606 (G); vicinity of Fort Sill, April 12, 1916, 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 403 Clemens 11727 (MBG); Cache, Comanche Co., dry hillsides, decomposed granite, July 19, 1917, Е. J. Palmer 12597 (MBG); Davis, Arbuckle Mts., April 1, 1916, Emig 399 (MBG). Texas: Comanche Springs, March, 1849, Lindheimer (С, MBG); Dallas, rocky prairies, April, 1875, Reverchon (G, NY, MBG); Dallas, dry uplands, March-June, year lacking, Rever- chon (а, MBG); “Upper Colorado," rocky places, 1847, Lind- һейтег 660 (С түре, MBG, US, Е); Fort Worth, rocky hill- sides, May 7, 1911, Ruth 241 (С, NY, ANSP, Е); “Witicha Mtns.,” July, 1852, Torrey (G, NY); data lacking, Lindheimer (ANSP, MBG); Dallas, dry soil, April-June, 1877, Reverchon 598 (US, MBG); Dallas, common in woods, May 7, 1900, Bush 646 (US, NY, MBG); Dallas, rocky prairies, June 30, 1877,“ Hall 515 (US, MBG, NY); Forks, May 27, year lacking, Rever- chon (MBG); Boerne, Kendall Co., low rocky creek banks, April 6, 1917, Е. J. Palmer 1147: (MBG); Dallas, rocky hills, West Dallas, June 22, 1899, Eggert (MBG); Hood Co., prairies, May 4, 1900, Eggert (MBG); Dallas, cement works, April 12, 1902, Reverchon (MBG); data lacking, Lindheimer 4 (MBG); Gillespie Co., date lacking, Jermy 145 (MBG); Dallas, open limestone hills, May 4, 1918, Е. J. Palmer 13496 (MBG); Lacey’s Ranch, Kerr Co., moist rocky creek banks, June 11, 1917, Е. J. Palmer 12233 (MBG); Bull Creek, near Austin, April 11, 1914, Young (MBG); Boerne, Kendall Co., moist rocky creek banks, April 20, 1917, Е. J. Palmer 11616 (MBG); Dallas, high prairies, April 12, 1902, Reverchon 3122 (MBG). 3. Amsonia elliptica (Thunb.) Roem. & Schult. Syst. Veg. 4: 432. 1819; A. DC. in DC. Prodr. 8: 384. 1844; Franch. & Savatier, Enum. Pl. Jap. 1: 315. 1874; K. Sch. in Engl. & Prantl, Nat. Pflanzenfam. 42: 143. 1895; Matsumura, Index РІ. Јар. 2: 505. 1912. Pl. 51, figs. 9-10. Tabernaemontana elliptica Thunb. Fl. Jap. 111. 1784. Ansonia elliptica (Thunb.) Raf. New Fl. Х. Am. 4: 58. 1838. “ Amsonia elliptica Sieb. & Гласс.” in Gray, Mem. Am. Acad. 6: 403. 18957. Herbaceous perennial from a slightly thickened root; stems 4-7 dm. tall, single or clustered from the base, erect or slightly [Vor. 15 404 ANNALS OF THE MISSOURI BOTANICAL GARDEN ascending, glabrous, or very slightly pubescent when young, branched above, the branches ascending or somewhat spread- ing; leaves alternate, relatively distant, the blades relatively narrow, lanceolate to linear-lanceolate, the lower 5-10 times аз long as broad, both the bases and the apices narrowly acute to acuminate, glabrous above, glaucescent beneath, becoming green in age; inflorescence loose, relatively few-flowered, pedicels 5-10 mm. long; calyx 1-2 mm. long, the lobes triangular-lanceo- late, glabrous; corolla salverform, the tube relatively broad, 10-12 mm. long, glabrous without; the lobes of about equal length, oblong-lanceolate, spreading; stigmatic-cap about as broad as tall, stigma depressed-capitate or truncate; follicles relatively stout, continuous, or very slightly torose, 4-6 cm. long, sessile, glabrous, 5-10-веедед; seeds 5-10 mm. long, oblong in outline, truncate at either end, variously pitted or wrinkled, brown. : Distribution: northern Japanese Archipelago. Specimens examined: JAPAN: Hakodate, 1861, Mazximowicz (С); Jesso, near Hako- date, 1861, Albrecht (G); Todahara, Musashi, May 24, 1891, Watanabe (G); Tokio, May 7, 1879, Matsumura (US); Musaski, Toda, May 27, 1911, collector lacking (US). ` 4. Amsonia Tabernaemontana Walt. Fl. Carol. 98. 1788; Pers. Syn. 1: 269. 1801; А. DC. in DC. Prodr. 8: 384. 1844; Rept. Torr. Bot. Mex. Bound. Surv. 159. 1859; Gray, Syn. Fl. М. Am. 2': 81. 1878; Wood, Classbook Bot. 589. 1860; Gat- tinger, Tenn. ГІ. 63. 1887; Coulter, Contr. U. 8. Nat. Herb. 2: 262. 1892; K. Schumann in Engl. & Prantl, Nat. Pflanzen- fam. 42: 143. 1895; Chapm. Fl. South. 0. S. 343. 1897; Rob- inson & Fernald in Gray, New Man. ed. 7, 661. 1908. Pl. 51, figs. 11-13. Anonymus suffrutez Gronov. Fl. Virg. ed. 2, 35. 1762. Tabernaemontana Amsonia L. Sp. РІ. ed. 2, 2: 301. 1762; Willd. Sp. Pl. 1?: 1246. 1798. Tabernaemontana humilis Salisb. Prodr. 148. 1796. - Amsonia latifolia Michx. Fl. Вог. Am. 1: 121. 1803; Pursh, Fl. Am. Sept. ed. 1, 1: 184. 1814; Roem. & Schult. Syst. Veg. 4: 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 405 432. 1819; Elliott, Sketch Bot. S. C. & Ga. 316. 1821; Darby, Bot. South. States, 434. 1860. Amsonia tristis Sm. in Rees, Cycl. 35: end of art. “Таһегпае- montana." 1819; А. DC. in DC. Prodr. 8: 384. 1844 Ansonia latifolia (Michx.) Raf. New Fl. N. Am. 4: 58. 1838. Amsonia Amsonia (L.) Britton, Mem. Torr. Bot. Club 5: 262. 1894; Britton & Brown, Ш. Fl. 3: 1. 1898; S. Coulter, Rept. Dept. Geol. Ind. 24: 880. 1899; Hitchcock, Fl. Kans. 13. 1899; Gattinger, Fl. Tenn. 137. 1901; Mohr, Contr. 0. 5. Nat. Herb. 6: 674. 1901; Small, Fl. Southeast. U. 5. 935. 1903; Lowe, Miss. State Geol. Surv. Bull. 17: 227. 1921. Herbaceous perennial from a thickened, slightly woody root; stems 3-10 dm. tall, usually clustered from the base, erect or slightly ascending, branched above, the branches ascending or spreading, occasionally somewhat pubescent when young; leaves alternate, relatively distant, ovate to oblong-elliptic, the bases of the lower obtuse to broadly acute, occasionally sparsely pubescent upon the lower surface when very young; inflores- cence relatively small and dense, barely held above the foliage, pedicels 3-5 mm. long; calyx 1-1.5 mm. long, glabrous, the lobes triangular-ovate; corolla salverform, the tube 6-8 mm. long, pubescent without, the lobes 4-6 mm. long, oblong to oblong-lanceolate, spreading; stigmatic-cap about as tall as broad, stigma depressed-capitate; follicles continuous, 8-10 cm. long, rather abruptly acuminate, sessile, glabrous, 5-15-веедед; seeds 5-11 mm. long, oblong in outline, truncate at either end, variously pitted and wrinkled, dark brown. Distribution: moist woods and waste-lands, river-banks, etc.; South Carolina, Tennessee, Illinois, eastern Missouri, centers, Oklahoma, eastern Kansas, көш наннан Arkansas, escaped from cultivation in Massachusetts, New Jersey, Pennsylvania, and Delaware. Specimens examined: MassAcHUSETTS: Boston, Back Bay waste-lands, Aug. 12, 1903, Williams (С); Hampden, June, 1911, Knowlton (NE). New Jersey: South New England Road, introduced in field, Cold Spring, Cape May Co., July 7, 1918, Brown (PBC). [Vor 15. 406 ANNALS OF THE MISSOURI BOTANICAL GARDEN DELAWARE: Wilmington, waste places, June 3-July 18, 1896, Commons (PBC). PENNSYLVANIA: Oakdale, near Philadelphia, June, 1863, Mar- tindale (PBC); Philadelphia, Broad Street & Germantown R. R., 1865, Martindale 4864 (MBG); Gradyville, Delaware Co., June 9, 1898, Painter (РВС); near Philadelphia, May, 1889, Leeds (РВС); Gradyville, June 3, 1904, Vail 546 (US). SouTH CAROLINA: Greenville Co., ravines near Caesar's Head, Aug. 5, 1881, J. D. Smith (G). TENNESSEE: Knoxville, thicket on Tennessee River bank, April and July, 1890, Ruth 174 (С); Knoxville, April, 1894, Ruth 466 (P, US); Knoxville, June, 1898, Ruth 480 (MBG). Пллхотв: Chandlersville, Aug. 19, 1886, Seymour 1584 (С, P). Missouri: St. Louis, July 2, 1895, Glatfelter (С, MBG); St. Louis, date lacking, Engelmann (С, MBG); Eagle Rock, un- common in barrens, June 22, Bush 11 (MBG, US); uncommon in rich woods, 4 miles e. of Carthage, May 27, 1906, E. J. Palmer 921 (MBG); Newton Co., cherty barrens, July 15, 1906, E. J. Palmer 12 (MBG); Carthage, rich woods, May 27, 1906, E. J. Palmer 818 (МВС); Noel, low ground, Мау 10, 1915, Bush 7513 (MBG); Noel, McDonald Co., thickets, hillsides, Sept. 12, 1913, Е. J. Palmer 4305 (MBG); Swan, common in woods, Oct. 4, 1899, Bush 753 (MBG). ARKANSAS: Fort Huron, date lacking, Edward (G); Fayette- ville, May, year lacking, Harvey 38 (ANSP); Fulton, low ground, April 17, 1905, Bush 2378 (MBG); Fayetteville, May 10, 1919, Wells (US). Т.ОТЛВГАМА: Hammond, April 10, 1889, Gallup 4 (US). OKLAHOMA: Leflore Co., Page, on bank of mountain creek near Rich Mountain, Sept. 8, 1913, Stevens 2670 (G, US); Page, Leflore Co., on rocky mountain-side, April 25, 1915, Blakely 8425 (G); Poteau, Leflore Co., July 13, 1915, E. J. Palmer 8286 (МВО). Kansas: Cherokee Со., rocky woods, Мау 7, 1897, Hitchcock 76a (MBG). 4a. Var. salicifolia (Pursh) Woodson, n. comb. Amsonia salicifolia Pursh, Fl. Am. Sept. ed. 1, 1: 184. 1814; 1928] WOODSON—STUDIES IN APOCYNACEAE. III 407 Roem. & Schult. Syst. Veg. 4:432. 1819; Elliott, Sketch Bot. S. C. & Ga. 316. 1821; A. DC. in DC. Prodr. 8: 384. 1844; Darby, Bot. South. States, 434. 1860; Wood, Classbook Bot. 589. 1860; Small, Fl. Southeast. U. 5. 935. 1903; Britton, Man. Fl. 737. 1907. | Ansonia salicifolia (Pursh) Raf. New Fl. М. Am. 4: 58. 1888. Herbaceous perennial from a slightly thickened root; stems 3-5 dm. tall, usually clustered from the base, erect or slightly ascending, glabrous or very slightly pubescent when young, branched above, the branches ascending or somewhat spread- ing; leaves alternate, the blades relatively narrow, lanceolate to linear-lanceolate, the lower 5-10 times as long as broad, both the base and the apex narrowly acute to acuminate, gla- brous above, glaucous or glaucescent beneath, becoming green in age; inflorescence loose, relatively few-flowered, pedicels 3-7 mm. long; calyx about 1 mm. long, the lobes minutely triangu- lar, glabrous; corolla salverform, the tube relatively narrow, 6-10 mm. long, scatteringly pubescent without, the lobes 5-7 mm. long, lanceolate, spreading; stigmatic-cap somewhat broader than tall, stigma depressed-capitate or truncate; follicles rela- tively slender, continuous or very slightly torose, 8-10-веедед; seeds 5-10 mm. long, oblong in outline, truncate at either end, variously pitted or wrinkled, brown. Distribution: river-banks and moist thickets generally; Vir- ginia, North Carolina, South Carolina, Georgia, Alabama, Louisi- ana, Kentucky, Tennessee, Indiana, Illinois, Missouri, Arkansas, and Texas. Specimens examined: Үткатхта: Petersburg, date lacking, Twomey (ANSP). Мовтн CAROLINA: Biltmore, river-banks, May 11, 1897, Bilt- more 81% (С, MBG, US, NY); same locality, Мау 2, 1896, Biltmore 81 (MBG); Weldon, April, 1897, Williamson (ANSP); same locality, April 19, 1908, Williamson (ANSP); Hot Springs, May 5, 1884, Smith (ANSP); Warm Springs, May 6, 1887, Smith (ANSP); Columbus, 1897, Townsend (US); Statesville, May, 1878, Hyams (US); Granville Co., May, 1873, Faxon (G); Mt. Tryon, Polk Co., moist rich soil on a level spot along a mountain rill, May, 1918, Millspaugh 4030 (Е); Weldon, April 19, 1908, Bartram (G); data lacking, Curtis (G). (Мог. 15 408 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN боотн CAROLINA: Oconee Co., Clemson College, low woods, April 16, 1906, House 1851 (US, NY). СвовотА: Thompson’s Mills and vicinity, Gwinnett Co., April 8, 1908, Allard 197 (US); Macon, date lacking, Green (ANSP); Stone Mountain, Yellow River, May 3, 1899, Sargent 69 (G). ALABAMA: Albertsville, April 22, 1899, Hosdy (US); Chicka- saw, rich places in the barrens, April, 1919, Graves 566 (US); Tuskaloosa, April, 1892, Ward (US); Auburn, Lee Co., April 22, 1900, Earle (G). Mississippi: Pass Christian, Мау 16, 1924, Cooper (MBG); Rolling Fork, April, 1895, Boyce (US). Louisiana: Feliciana [East or West?] Parish, Carpenter (С); east of Baton Rouge, April 20, 1874, Joor (F). INDIANA: banks of the Wabash River, June, 1868, Allen (Е). Kentucky: Bowling Green, 1903, Price (MBG); barrens of Kentucky River near Hopkinsville, date lacking, Buckley (MBG). TENNESSEE: Franklin Co., May 5, 1898, Eggert (MBG); Nash- ville, 1880, Hubbard 2268 (G). Палхо1в: exact locality lacking, 1845, Mead (С); Grantsburg, April 28, 1900, collector lacking (P); Conologue, April, 1924, Woodson (MBG). Missourr: bank of Meramec River near Windsor Springs, April 19, 1891, Douglass (US); Cave Spring, 1887, Blankinship (US); Cave Spring, Greene Co., June 18, 1905, Standley (US); Tyson, St. Louis Со., Мау 19, 1918, Drushel 3757 (MBG); banks of the Meramec River, Minke, St. Louis Co., May 17, 1919, Greenman 3944 (MBG); Chadwick, May 15, 1907, Bush 4461 (MBG). ARKANSAS: Baker Springs, Howard Co., April 10, 1909, Kel- logg (MBG); data lacking, Pitcher (ANSP). Texas: Beaumont, low wet woods, March 16, 1918, E. J. Palmer 13090 (MBG). 4b. Var. Gattingeri Woodson, п. уаг.! Herbaceous perennial from a thickened slightly woody root; ‘Var. Gattingeri var. nov., plus-minusve pilosa varietatem genuinam simulans differt foliis longioribus basi acutis; corollae tubo lanoso.—Tennessee, Nashville, June, year lacking, A. Gattinger (Gray Herb. TYPE). 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 409 stems 3-10 dm. tall, pubescent, becoming glabrous, somewhat clustered at the base, erect or ascending, branched above, the branches ascending or spreading; leaves relatively distant, alter- nate, the blades lanceolate to linear-lanceolate above, the lower 5-10 times аз long as broad, both base and apex narrowly acute to acuminate, green, pubescent, frequently densely so, becoming glabrous in age; inflorescence compact, many-flowered, pedicels 2-4 mm. long; calyx 2-4 mm. long, the lobes narrowly triangu- lar, glabrous, or with a few scattered hairs; corolla salverform, the tube 7-10 mm. long, densely pubescent or villous without, especially in the sinuses of the lobes, the lobes 5-8 mm. long, lanceolate, spreading; stigmatic-cap much broader than tall, stigma truncate; follicles slender, continuous or very slightly богове, 9-14 cm. long, acuminate, sessile, glabrous, 7-11-seeded; seeds 5-12 mm. long, oblong in outline, truncate or slightly tapered at the ends, variously pitted or wrinkled, brown. Distribution: woods and ravines, northern Georgia, Tennessee, Illinois, Missouri, southeastern Kansas, eastern Oklahoma, and northeastern Texas. Specimens examined: GEORGIA: Jasper City, 1847, Porter (G). TENNESSEE: Nashville, June, year lacking, Gattinger (G TYPE, MBG); Nashville, islands in Cumberland River, pope 1878, Gattinger (MBG, NY, ANSP, F). Kentucky: barrens of the Kentucky River, exact locality lacking, 1860, Short (MBG). Пллхотв: Athens, 1861, Най (С, P, MBG); Olney, Richmond Co., Turkey Creek bottoms, May 19, 1914, Ridgway 104 (G, МВС); Grantsburg, April 28, 1900, Baker (P); East Hannibal, June 6, 1913, Davis 398 (MBG, US); along road west of Fish Lake, St. Clair Co., July 16, 1898, Norton (MBG); damp shady thickets, American Bottoms, opposite St. Louis, May, 1845, Engelmann (MBG); Queens Lake, Clinton Co., May 20, 1917, Ledman (MBG); Venedy, May 18, 1926, Anderson & Woodson 5 (MBG); Conologue, May 16, 1926, Woodson & Stevenson 41 (МВО). Missouri: Alba, rich bluff woods, April 29, 1909, Е. J. Palm- er 1819 (б, MBG); St. Louis, July 28, 1910, Sherff 801 (С, Е, (Мог. 15 410 ANNALS OF THE MISSOURI BOTANICAL GARDEN MBG); Webb City, gravelly branches, Sept. 2, 1909, Е. J. Palmer 2620 (G, MBG); Winfield, Lincoln Co., June 7, 1916, Davis 1403 (MBG); Bower’s Mill, Lawrence Co., rich hill-side woods, April 22, 1908, Е. 7. Palmer (MBG); Allenton, June 10, 1884, Kellogg (MBG); Elmont, Мау 23, 1914, Emig (MBG); Gascondy, July 21, 1914, Emig 221 (MBG); Gray’s Summit, May 15, 1926, Greenman 4493 (MBG); Allenton, June, 1880, Letterman (MBG, US). ARKANSAS: Benton Co., date lacking, Plank (MBG); Eureka Springs, April 27, 1899, Trelease (MBG); Little Rock, May, 1886, Hasse (Е). OKLAHOMA: Miami, on dry bank of draw, Aug. 26, 1913, Stevens 2337 (G, MBG, US); Page, on rocky mountain-side, April 25, 1915, Buckley 3425 (G, MBG); rocky hills, Wichita Mts. not common, July, 1891, Sheldon 224 (MBG). 5. Amsonia ludoviciana Vail in Small, Fl. Southeast. U. 5. ed. 2, 935. 1913. Herbaceous perennial from a slightly thickened woody root; stems 5-11 dm. tall, pubescent, at least when young, sparingly branched, erect or ascending, the branches erect or ascending; leaves relatively distant, alternate, the blades elliptic, both base and apex acute to acuminate, 5-8 cm. long, essentially glabrous above, densely white-lanose beneath, pedicels 2-4 mm. long; inflorescence relatively dense, several-flowered, pedicels 2-4 mm. long; calyx 2-3 mm. long, the lobes triangular, 1-1.5 mm. long, pubescent; corolla salverform, the tube 5-9 mm. long, densely pubescent or villous without, the lobes about equalling, or slightly exceeding, the tube, lanceolate, spreading; stigmatic-cap about as broad as tall, stigma truncate; follicles slender, continuous, 8-10 cm. long, acuminate, sessile, manifestly pubescent, 6-10- seeded; seeds 5-12 mm. long, oblong-ovoid in outline, truncate at the ends, variously pitted and wrinkled, dark brown. Distribution: known only from southern Louisiana. Specimens examined: LovistANA: New Orleans, date lacking, Ingalls (МУ); Shacky- nody, April, year lacking, Hale (NY). 1928] WOODSON—STUDIES IN APOCYNACEAE. III 411 ЗовсЕмоз П. Өрнімстовірном (К. Schumann) Woodson Subgenus П. 8рнгхстовтрнох (К. Schumann) Woodson, п. comb. §Sphinctosiphon K. Schumann in Engl. & Prantl, Nat. Pflan- zenfam. 42: 143. 1895; Dalla Torre «с Harms, Gen. Siph. 406. 1904. Bracteoles conspicuous, giving the inflorescence a chaffy ap- pearance; mouth of the corolla-tube markedly constricted at anthesis; stigma apiculate by two distinct obtuse lobes; follicles continuous, not articulated, fibrous, not horny in texture; seeds oblong in outline, truncate at either end, variously pitted and wrinkled; plants of the southwestern United States and north- ern Mexico. Spp. 6-13 Section I. Міснахтнак Woodson. Corolla-tube 1-1.5 ст. long; calyx 1-4 mm. long; follicles 4-7 cm. long; seeds 4-8 mm. long. KEY TO THE SPECIES a. Follicles slender; seeds fertile. b. Corolla-lobes 3-6 mm. long, ovate or oblong. с. Corolla-lobes 3-4 mm. long; plant entirely glabrous. ... . 6. А. Palmeri сс. Corolla-lobes 5-6 mm. long; plant pubescent, at 0 the calyx- d. Stem and leaves glabrous; pedicels 3-5 mm. long; inflores- cance 1008... 6. : css toned cee на эх» 7. A. pogonosepala dd. Stem and leaves pubescent; рде 1-2 mm. long, or ргас- tically sessile; inflorescence dens e. Calyx glabrous; inflorescence many-flowered.. ay A. hirtella e. Calyx pubescent; infl flowered. . .9. А. Standleyi Бр. Pra 6-8 mm. long, В... do A. latifolia ah. Follicbe tort; seeds ат е... 125 ее dao 11. A. Kearneyana 6. Amsonia Palmeri Gray, Proc. Am. Acad. 12: 64. 1877; Gray, Syn. Fl. N. Am. 2:: 82. 1878. Pl. 52, figs. 14-15. Amsonia Fremontii Rydb. Bull. Torr. Bot. Club 40: 465. 1913. nomen. Herbaceous perennial from а somewhat thickened root, gla- brous; stems 3-5 dm. tall, usually clustered from the base, erect or slightly ascending, sparingly branched above, the branches ascending; leaves alternate, relatively numerous, oblong-lanceo- late to linear-lanceolate above, the blades 2.5-7 cm. long, 4-8 mm. broad; inflorescence relatively few-flowered and loose, held well above the foliage; pedicels 1-3 mm. long, or practically [Vor. 15 412 ANNALS OF THE MISSOURI BOTANICAL GARDEN lacking; calyx 3-4 mm. long, sparsely hairy, the lobes subulate; corolla salverform, the tube constricted at the mouth, 1-1.8 em. long, the lobes ovate to ovate-oblong, 3-4 mm. long, erect or spreading; stigma apiculate by two distinct obtuse lobes; follicles 4—6 cm. long, acuminate, sessile, glabrous, continuous, 5-10-seeded; seeds 4-8 mm. long, oblong in outline, truncate at either end, variodaly pitted or wrinkled, chocolate-brown. Distribution: Ал» апа Хеу Мехко. Specimens examine New Mexico: nad locality lacking, 1851-52, Wright 1669 (С түре, MBG). ARIZONA: exact locality lacking, 1884, Lemmon 3248 (G); 50 miles s. of Lee's Ferry, June 12, 1890, M. E. Jones (P, US); Hillside, May 1, 1903, alt. 3700 ft., Jones (MBG); Beale's Spring, date lacking, Lemmon & Lemmon (US); exact locality lacking, 1887, Mearns 152 (NY). 7. Amsonia pogonosepala Woodson, n. sp.! Herbaceous perennial from a thickened woody root; stems 5-8 dm. tall, glabrous, clustered from the base, erect or slightly ascending, freely branched above, the branches ascending or spreading; leaves alternate, relatively numerous, glabrous, lance- olate to oblong-lanceolate, the blades 1-1.5 ст. broad, 5-7 cm. long, acute to acuminate at both base and apex, petiolate, the petioles 1-3 mm. long; inflorescence loose, relatively many- flowered; pedicels 2-4 mm. long; calyx 3-6 mm. long, the lobes subulate, conspicuously ciliate, 2-5 mm. long; corolla salverform, the tube constricted at the orifice, 12-15 mm. long, glabrous without, the lobes 5-6 mm. long, ovate to oblong, spreading; stigma apiculate by two distinct obtuse lobes; follicles 1.2-8 cm. long, acuminate, sessile, glabrous, continuous, 4—15-seeded; seeds 7-10 mm. long, oblong-truncate in outline, variously pitted or wrinkled, reddish brown. 1 Amsonia pogonosepala sp. nov., humila saepe basaliter ramosa 5-8 dm. alta; ramis erectis vel laxe ascendentibus glabris; foliis lanceolatis oblongo-lanceolatis petiolatis 5-7 ст. longis 1-1.5 em. latis glabris; lobis calycis piloso-ciliatis subulatis 2-5 mm. longis; corollae lobis ovatis vel ovato-oblongis 5-6 mm. longis tubo sub- clavato dimidio brevioribus distendatis; stigmate subtrochleari apice bilobato; fol- liculis teretibus gracilibus continuis glabris sessilibus 2-8 em. longis.—Arizona, dry rocky hills, San Francisco Mts., April, 1881, Н. H. Rusby 256 (MBG түре). 1928] WOODSON—STUDIES IN APOCYNACEAE. III 413 Distribution: southern Arizona. Specimens examined: Arizona: dry rocky hills, San Francisco Mts., April, 1881, Rusby 256 (МВС түре, ANSP, NY); small sandy wish between Apache Junction and Canyon Lake, June 21, 1928, Harrison & Peebles 5540 (MBG, US); near Mormon Flats, April 1, 1928, Peebles, Harrison, & Kearney 3820 (05). 8. Amsonia hirtella Standley, Proc. Biol. Soc. Wash. 26: 118. 1913; Wooton & Standley, Contr. U. S. Nat. Herb. 19: 505. 1915. Herbaceous perennial from & somewhat thickened root, hir- tellous; stems 3-5 dm. tall, usually clustered from the base, erect or somewhat ascending, very sparingly branched above, the branches ascending; leaves alternate to subverticillate above, relatively numerous, lanceolate to linear-lanceolate, the blades 3-5 cm. long, 2-5 mm. broad, sessile to subsessile; inflorescence dense, relatively many-flowered; pedicels 1-2 mm. long or prac- tically lacking; calyx 4-5 mm. long, glabrous, except for a few scattered hairs at the tips, the lobes subulate; corolla salverform, the tube constricted at the mouth, 12-15 mm. long, glabrous without, the lobes 5-6 mm. long, ovate to ovate-oblong, slightly spreading; stigma apiculate by two distinct obtuse lobes; fol- licles unknown. Distribution: known only from southwestern New Mexico. Specimens examined: New Mexico: Grant Co., cafions, May 1, 1892, Mearns 117 (US TYPE). 9. Amsonia Standleyi Woodson, n. гр: Pl. 52, figs. 16-17. Herbaceous perennial from а somewhat thickened woody root, densely pubescent; stems 3-5 dm. tall, usually clustered from the base, erect or slightly ascending, freely branched above, the ! Amsonia Standleyi sp. nov., pilosa humila saepe basaliter ramosa 3-5 dm. alta; ramis erectis ~ laxe ascendens folis lanceolatis linearibusque plerumque m. longis 4-10 mm. latis alternis vel subverticillatis; lobis calycis pilosis Ёс 16 4-5 mm. longis; corollae lobis ovatis vel ovato-oblongis 5-6 mm. longis tubo subclavato dimidio brevioribus distendatis; stigmate sub- trochleari apice bilobato; folliculis teretibus gracilibus continuis glabris sessilibus 6-7 cm. longis.—New Mexico, 1851-52, C. Wright (Gray Herb. TYPE). [Vor. 15 414 ANNALS OF THE MISSOURI BOTANICAL GARDEN branches ascending or slightly spreading; leaves alternate to subverticillate above, relatively numerous, lanceolate to linear- lanceolate, the blades 3—7 ст. long, 4-10 mm. broad, narrowed to an inconspicuous petiole, or practically sessile; inflorescence dense, relatively few-flowered; pedicels 1-2 mm. long, or prac- tically lacking; calyx 4—5 mm. long, densely pubescent through- out, the lobes subulate; corolla salverform, the tube constricted at the mouth, 8-10 mm. long, glabrous without, the lobes ovate to ovate-oblong, 5-6 mm. long, spreading; stigma apiculate by two distinct obtuse lobes; follicles 6-7 cm. long, acuminate, sessile, glabrous, continuous, 5-10-seeded; seeds 4-8 mm. long, oblong in outline, truncate at either end, variously pitted and wrinkled, brown. Distribution: и New Mexico, and Chihuahua. Specimens examine UNITED STATES: Texas: Bofecillos, May 18, 1881, Havard (US). | Кем Mexico: exact locality lacking, 1851-52, Wright (С TYPE). Mexico: CHIHUAHUA: Candelaria, Oct. 24, 1911, Stearns 228 (US). This species is named in honor of Mr. Paul C. Standley, who provisionally referred the Havard and Stearns specimens, bear- ing fruit only, to A. hirtella, in describing that species, but foresaw that when the flowers corresponding to the fruit should be found they would constitute a new species. 10. Amsonia Jonesii Woodson, new name. Amsonia latifolia M. E. Jones, Contr. West. Bot. 12: 50. 1908, not Michx.; Rydb. Fl. Rocky Mts. 668. 1917; Tidestrom, Contr. U. S. Nat. Herb. 25: 418. 1925. Amsonia texana Rydb. Fl. Rocky Mts. 668. 1917, not Heller. Herbaceous perennial from a thickened, frequently very woody root, glabrous; stems 2-4 dm. tall, usually much clustered from the base, erect or ascending, sparingly branched, the branches ascending; leaves alternate, numerous, ovate to ovate-oblong, glaucous, the blades 3-5 cm. long, 1-2 cm. broad, petiolate; ‘inflorescence dense, relatively many-flowered; pedicels 3-5 mm. 1928] WOODSON—STUDIES IN APOCYNACEAE. III 415 long; calyx about 1.5 mm. long, the lobes triangular; corolla salverform, 5-8 mm. long, the lobes narrowly oblong-lanceolate, 5-7 mm. long, spreading, slightly pubescent at the tips when in aestivation; stigma apiculate by two distinct obtuse lobes; follicles 5-8 cm. long, acuminate, sessile, glabrous, continuous, 4—8-seeded; seeds 4-6 mm. long, oblong in outline, transversely truncate at either end, variously pitted and sculptured, brown. Distribution: rocky gorges and canons, southwestern Colorado, southeastern Utah, and northeastern Arizona. Specimens examined: СотовлАро: Grand Junction, alt. 4500 ft., June 21, 1894, M. E. Jones 5469 (P); Grand Junction, alt. 4000 ft., May 28, 1895, M. E. Jones (P); McElmo Creek, Montezuma Co., July 19, 1895, Eastwood 72 (G, MBG); Grand Junction, June 21, 1894, Jones 5476q (MBG, ANSP, US, NY). Uran: Monroe, Sevier Co., alt. 5000 ft., May 24, 1894, М. Е. Jones 6446 (Р түре, MBG). ARIZONA: Navajo Wells, alt. 5000 ft., Мау 24, 1894, M. Е. Jones 5289aa (Р, MBG); Pagumpa, alt. 4000 ft., April 21, 1894, М. Е. Jones 5093 (P, US, NY, MBG). 11. Amsonia Kearneyana Woodson, п. sp.! Herbaceous perennial from a thickened, somewhat woody root, more or less pilose; stems 4-8 dm. tall, usually clustered from the base, erect or ascending, sparingly branched, the branches ascending; leaves alternate to subverticillate, oblong-lanceolate to lanceolate, 4-7 cm. long, 1–1.5 ст. broad, petiolate, or sub- sessile; inflorescence dense, many-flowered; pedicels 1 mm. long or practically lacking; calyx 3-5 mm. long, the lobes subulate- aristate, densely pilose-ciliate; corolla salverform, the tube con- stricted at the orifice, 1-1.2 ста. long, the lobes 3—5 mm. long, oblong to ovate, erect or slightly spreading; stigma apiculate by two distinct obtuse lobes; follicles short and obviously de- 1 Amsonia Kearneyana sp. nov., plus-minusve pilosa basaliter ramosa 4-8 dm. alta; ramis ascendentibus; foliis авгай үе! subverticillatis subsessilibus oblongo- lanceolatis 4-7 cm. longis 1-1.5 cm. latis; lobis calycis subulato-aristatis 3-5 mm. longis; corollae tubo longo subclavato 1-1.2 ст. longo lobis ovatis 8-5 mm. longis erectis vel ascendentibus; stigmate subtrochleari apici bilobato; folliculis deformis; seminibus sterilibus.—Arizona, Pima Со., South Сайоп, April 9, 1928, Е. Thackery 55 (MBG түре). [Vor. 15 416 ANNALS OF THE MISSOURI BOTANICAL GARDEN formed but essentially continuous, 2-5 cm. long, 5-7 mm. broad; seeds sterile. Distribution: southern Arizona. Specimens examined: ARIZONA: South Canyon, Baboquivari Mts., May 24, 1926, Thackery 2018 (US, MBG); South Canyon, Baboquivari Mts., March 29, 1927, Peebles, Harrison & Kearney 3820 (US, MBG); South Сайоп, Pima Co., April 9, 1928, T'hackery 55 (MBG TYPE). Because of its appearance intermediate between А. Siandleyr or A. Palmeri and A. brevifolia or A. tomentosa, because of its geographical position, and because of its complete sterility, A. Kearneyana is regarded as а natural hybrid between the sub- genera Sphinctosiphon and Атисшата. The flowers are decidedly of the type of A. Palmeri, with oblong-ovate, erect or ascending corolla-lobes, while the broad foliage is very similar to that of A. brevifolia or A. tomentosa. However, since several colonies have been found in the same general vicinity, it is thought better to consider it as a distinct species in the light of recent opinions concerning the origin of species by means of hybridization. Since it demonstrates its recent creation by its sterility and its irregular pilosity, it should probably be considered the most recently evolved species of the genus Amsonia. A. Kearneyana is so named in honor of Mr. T. H. Kearney, of the United States Bureau of Plant Industry, who brought the plant to the attention of the author, and furnished much valuable information regarding the genus in Arizona. Section и. Lonarrtorae Woodson. Corolla-tube 3-4 ст. long; calyx 4-8 mm. long; follicles 7-9 ст. long; seeds 5-12 mm. long. Кет то THE SPECIES а. Plant glabrous; corolla-lobes 11-13 mm. long................ 12. A. longiflora aa. Plant pubescent; corolla-lobes 5-7 mm. long............. 13. A. salpignantha 12. Amsonia longiflora Torr. Bot. Mex. Bound. Surv. 159. 1859; Gray, Syn. Е. ЇЧ. Am. 2: 82. 1878; Hemsley, Biol. Cent.-Am. Bot. 2: 308. 1881; Wooton & Standley, Contr. U. 8. Nat. Herb. 19: 504. 1915. Pl. 52, figs. 18-20. Herbaceous perennial from a thickened somewhat woody root, 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 417 glabrous; stems 3.5-6 dm. tall, usually clustered from the base, erect or ascending, copiously branched at maturity, the branches ascending or spreading; leaves alternate to subverticillate, linear- lanceolate to filiform, 2.5 cm. long, 1-2 mm. broad, sessile; in- florescence relatively loose, usually containing only 5-10 flowers; pedicels 2-5 mm. long; calyx 6-8 mm. long, the lobes subulate- aristate; corolla trumpet-shaped, the tube constricted at the mouth, 3-4 cm. long, the lobes 11-13 mm. long, oblong-lanceo- late, spreading; stigma apiculate by two distinct obtuse lobes; follicles slender, continuous, 7-9 cm. long, acuminate, sessile, 5-15-seeded; seeds 5-10 mm. long, elliptic-oblong in outline, truncate at either end, variously pitted or wrinkled, brown. Distribution: southeastern New Mexico, extreme western Texas, and north-central Mexico. Specimens examined: UNITED STATES: Texas: El Paso, 1881, Vasey (С, US, MBG); Hood Co., dry rocky prairie, Sept. 5, 1903, Reverchon 3881 (MBG); El Paso, April, 1852, Parry (MBG); El Paso, rocky ravines, Wright 1168 (С, NY түре, MBG); data lacking, Wright 72 (NY). New Mexico: base of Sacomento Mt., Alamogordo, April 14, 1902, Rehn & Viereck (ANSP); in arroyo, base of foothills, Alamogordo, May 19, 1902, Rehn & Viereck (ANSP); Rio Gila, Aug. 15, 1902, Wooton (US). ARIZONA: Sonoika Creek, south of Patagonia, April 15, 1908, Tidestrom 848 (US). ЕХІСО: DuRANGO: vicinity of the city of Durango, April-Nov., 1896, E. Palmer 90 (G, MBG, NY). 13. Amsonia salpignantha Woodson, n. sp. РІ. 52, figs. 21-22. Herbaceous perennial from a thickened somewhat woody root, 1 Amsonia salpignantha sp. nov., pilosa vel scabra basaliter ramosa 2-3.5 dm alta; ramis erectis vel ascendentibus; foliis multis alternis vel subverticillatis ses- silibus lineari-lanceolatis filiformibusque 2-5 cm. longis .5-4 mm. latis; lobis calycis lineari-lanceolatis 4-5 mm. longis; corollae tubo longo subclavato 3-4 cm. longo; corollae lobis ovatis 5-7 mm. longis distendatis; stigmate subtrochleari apici bilo- bato; folliculis teretibus gracilibus continuis glabris sessilibus 7-9 om. longis.— Texas, Hamilton Co., rocky prairies on the Cowhouse Creek, May, 1884, J. Rever- chon 1557 (MBG TYPE). [Vor. 15 418 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN pubescent, scabrous in age; stems 2-3.5 dm. tall, usually clus- tered from the base, erect or ascending, branched, the branches ascending; leaves numerous, alternate to subverticillate, linear- lanceolate to filiform, 2-5 ст. long, .5-4 mm. broad, sessile; inflorescence relatively dense, containing usually 10-30 flowers; pedicels 1-4 mm. long, or practically lacking; calyx 4-5 mm. long, the lobes subulate or narrowly lanceolate; corolla trumpet- shaped, the tube constricted at the mouth, 3-4 cm. long, gla- brous without, the lobes 5-7 mm. long, oblong-ovate, spreading; stigma apiculate by two distinct obtuse lobes; follicles slender, continuous, 7-9 em. long, acuminate, sessile, 5-15-seeded; seeds 5-12 mm. long, oblong in outline, truncate at either end, vari- ously pitted or wrinkled, brown. Distribution: southwestern Texas and Chihuahua. Specimens examined: UNITED STATES: Texas: Hamilton Co., 1885, Reverchon 99 (С, MBG); exact locality and date lacking, Pope (G); rocky prairies on the Cow- house Creek, Hamilton Co., May, 1884, Reverchon 1557 (F, MBG түре); Limpio Mts., 1883, Havard (US); Austin, 1880, Oberwetter (US); Del Rio, Dec. 7, 1891, Plank (NY). Mexico: СНТНУОАНУА: exact locality lacking, 1852, Wright 1671 (С). Зоваемов III. AnTICULARIA Woodson Subgenus ПІ. Arricutanra Woodson, n. subgen. §Sphinctosiphon K. Schumann in Engl. & Prantl, Nat. Pflan- zenfam. 42: 143. 1895, in part; Dalla Torre & Harms, Gen. Siph. 406. 1904, in part. Bracteoles conspicuous, giving the inflorescence a somewhat chaffy appearance; mouth of the corolla-tube markedly con- tracted in anthesis; stigma apiculate by two distinct obtuse lobes; follicles torose, articulated into thickish constricted seg- ments, horny, not fibrous in texture; seeds elliptic in outline, rounded or pointed at the ends, rarely truncate, relatively smooth and corky; plants of the southwestern United States and north- ern Mexico. Spp. 14-17. 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 419 KEY TO THE SPECIES a. Plant glabrous, or with but scattered hairs b ча ovate to ovate-lanceolate они corolla-tube about 10 mm. ОД ins Lane + 45 оа skew a свет eee 14. А. brevifólia bb. Rd lanceolate to linear above; corolla-tube about E. mm. long. Sea da рози a РЕН икра - алх 2. . А. Eastwoodiana аа. Plant villous or tomentose. : b. Leaves ovate to ovate-lanceolate, ребојаје............. 16. A. tomentosa bb. Leaves lanceolate to linear, sessile or subsessile........... 17. А. arenaria 14. Amsonia brevifolia Gray, Proc. Am. Acad. 12: 64. 1877; Gray, Syn. Fl. №. Аш. 21:81. 1878; Watson, Bot. Cal. 2: 462. 1880; Coville, Contr. U.S. Nat. Herb. 4: 142. 1893; Wooton & Standley, Contr. U. S. Nat. Herb. 19: 505. 1915; Rydb. Fl. Rocky Mts. 668. 1917; Davidson & Moxley, Fl. South. Cal. 278. 1923; Tidestrom, Contr. U. S. Nat. Herb. 25: 418. 1925; Jepson, Man. Fl. Pl. Cal. 768. 1925. Pl. 53, figs. 26-28. Herbaceous perennial from a thickened fibrous root, glabrous; stems 1.5-3.5 dm. tall, usually clustered from the base, erect or ascending, sparingly branched; leaves alternate, numerous, ovate- oblong to oblong-lanceolate, 2-3 cm. long, .5-1.5 ст. broad, acute to acuminate at either end; inflorescence dense, much divided at maturity; pedicels .5-1.5 mm. long or practically lacking; calyx 2-4 mm. long, the lobes subulate, bluish tinted; corolla salverform, the tube constricted at the mouth, 7-10 mm. long, the lobes ovate to ovate-oblong, 4—6 mm. long, spread- ing; stigma apiculate by two distinet obtuse lobes; follicles 5-7 em. long, torose, articulated into thickish constricted segments, sessile, glabrous, 3-10-seeded; seeds 5-7 mm. long, elliptic- lanceolate in outline, sharply truncate at one or both ends, never tapering sharply, 8-10 mm. long, 3.5-4 mm. broad, smooth or slightly wrinkled, yellowish-brown. Distribution: deserts and mountain slopes, southwestern Utah, northwestern Arizona, southern Nevada, and southern Cali- fornia. Specimens examined: Отан: Kanab, Logan Co., 1872, Thompson (С); exact locality and date lacking, 1874, Parry (G); Garfield Co., 1883, Siler (ANSP). ARIZONA: Mokiah Pass, 1877, E. Palmer 302 (С TYPE). [Vor. 15 420 ANNALS OF THE MISSOURI BOTANICAL GARDEN Nevapa: Eldorado Cañon at Nelson, alt. 3000 ft., April 30, 1907, М. Е. Jones (Р); 22 miles south of Searchlight, March 26, 1924, Jaeger (P); Las Vegas, June, 1915, K. Brandegee (P); Ashmeadows, alt. 3000-4000 ft., May-Oct. 1898, Ригриз 5988 (P); Cottonwood Springs, Las Vegas Valley, April 30, 1891, Bailey 1885 (US). CALIFORNIA: Mojave region, June, 1876, Е. Palmer 435 (С, MBG, ANSP, US); Mojave Desert near San Bernardino, 1880, Lemmon (G); Colorado Desert, in desert sands, April 24, 1921, Spencer 1778 (G); San Bernardino, May, 1882, Parish 1332 (G, MBG, US); San Bernardino Co., north slope of San Ber- nardino Mts., alt. 4000-6000 ft., June 15, 1895, Parish 3765 (G); Hesperia, in desert sand, Mojave Desert, alt. 3100 ft., May 8, 1917, Spencer 347 (G, P); Kelso, alt. 3000 ft., May 2, 1906, M. E. Jones (P); Keyes’ Ranch, alt. 3500 ft., common along a wash, May 7, 1922, Munz & Johnston 6258 (Baker); Corn Springs, rocky slope, high gorge, alt. 2500 ft., Munz & Keck 4843 (Baker); Goffs, Mohave Desert, March 28, 1924, Jaeger (P); entrance to Deep Creek, slope of San Bernardino Mts., alt. 3500 ft., Мау 9, 1921, Jaeger 288 (Baker); Quail Springs, Morango Pass, alt. 4000 ft., April 30, 1921, Munz 4585 (Baker); Cactus Flat, San Bernardino Co., alt. 6000 ft., June 25, 1926, Munz 10505 (Baker); Cotton-wood Springs, E. Riverside Co., March 26, 1926, Jaeger (P); Cushenberry Canyon, San Bernardino Co., June 1, 1892, Parish 2411 (F); Willow Creek Mt., Panamint Mts., May 22, 1891, Coville & Funston 825 (US); Mojave Desert, April-May, 1906, Saunders (ANSP); Cot- tonwood Pass, Riverside Co., May, 1905, Hall 6006 (05), same locality, April 12, 1924, Evermann (M BG). As an instance of the manner in which specific criteria have been applied in the present paper, the case of the specific indi- viduality of А. brevifolia Gray and A. tomentosa Тогг. & Frém. may be of interest, particularly to southwestern botanists. The two species mentioned are both members of the subgenus Ar- licularia and inhabit portions of southern California, southern Nevada, and southwestern Utah, and are seldom found sepa- rately. Јерѕоп,! іп 1925, came to the decision that they rep- ‘Jepson, W. L. Man. КІ. Pl. Cal. 768. 1925. 1928) WOODSON—STUDIES IN APOCYNACEAE. III 421 resented variation only, and reduced A. tomentosa to a variety of A. brevifolia, although if either should be reduced, the former should remain since it antedates the latter by thirty-two years. The important difference between the two species is the remark- able pubescence of A. tomentosa, a character which is not known to vary, judging from the copious herbarium material which the writer has been privileged to examine. А. tomeniosa is always remarkably pubescent, even to the mature follicles, while A. brevifolia is always found to be completely glabrous. More- over, the seeds of the two species are distinct, a fact which is evidently little appreciated. The seeds of A. brevifolia are sharply truncate at one or both ends and never taper sharply, measuring 8-10 mm. long and 3.5-4 mm. broad. The seeds of A. tomentosa taper decidedly at both ends and are slightly arcuate, measuring from 12 to 13 mm. long and 3 to 4 mm. broad. Illustrations of the seeds are to be found in pl. 53. In the presence of the seed difference and the non-intergrada- tion of the pubescence or glabrousity of the two species, it has been thought advisable to treat the species as representing a striking affinity rather than as varieties. 15. A. Eastwoodiana Rydb. Bull. Torr. Bot. Club 40: 465. 1913; Rydb. Fl. Rocky Mts. 668. 1917; Tidestrom, Contr. Т). $. Nat. Herb. 25: 418. 1925. Herbaceous perennial from a thickened fibrous-woody root, glabrous; stems clustered from the base, 3-5 dm. tall, erect or ascending, branched from near the base, the branches ascending or spreading; leaves alternate, rather distant, oblong-lanceolate below to linear-lanceolate above, 3-5 cm. long, .3-1 cm. broad, acute at either end, subsessile; inflorescence relatively small, loose; pedicels 4-7 mm. long; calyx 2-2.5 mm. long, the lobes subulate; corolla salverform, the tube constricted at the mouth, 1-2 спа. long, the lobes 4-6 mm. long, oblong, spreading; stigma apiculate by two distinct obtuse lobes; follicles 5-8 cm. long, torose, articulated into thickish constricted segments, sessile, glabrous, 3-5-seeded; seeds elliptic in outline, tapered at опе or both ends, 14-15 mm. long, 4-5 mm. broad, smooth or slightly wrinkled, reddish brown. [Vor. 15 422 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN Distribution: stream margins and ravines, Utah and Arizona. Specimens examined: Отан: San Juan Co., Willow Creek, July 14, 1895, Eastwood 73 (G, MBG); San Rafael Swell, Emery Co., May 12, 1914, М. Е. Jones (P); Moab, June 6, 1913, М. Е. Jones (P); 10 miles east of Holbrook, June 22, 1901, Ward (NY түре, US). ARIZONA: Lee's Ferry, June 13, 1890, M. E. Jones (Р, MBG); Kayenta, 1922, Weatherill (МУ). 16. Amsonia tomentosa Torr. & Frém. in Frém. Rept. 1843- 44, 316. 1845; Walpers, Ann. Bot. Syst. 1: 504. 1849; Rept. Torr. Bot. Mex. Bound. Surv. 158. 1859; Gray, Syn. Fl. №. Am. 2: 81. 1878; Hemsley, Biol. Cent.-Am. Bot. 2: 308. 1881; Rydb. Fl. Rocky Mts. 668. 1917; Davidson & Moxley, ЕЙ]. South. Cal. 278. 1923; Tidestrom, Contr. U. S. Nat. Herb. 25:418. 1925. Pl. 53, figs. 23-25. Amsonia brevifolia Gray var. tomentosa (Тогг. & Frém.) Jep- son, Man. Fl. Pl. Cal. 768. 1925. Herbaceous perennial from a slightly woody root, densely tomentose; stems 3-4 dm. tall, usually clustered at the base, erect or ascending, branched, the branches ascending or spread- ing; leaves alternate, rather numerous, ovate-oblong below to oblong-lanceolate above, 2-4 em. long, 1-1.5 ст. broad, acute at either end, the apices of the upper leaves conspicuously at- tenuate; inflorescence small, very dense, usually held well above the foliage; pedicels .5-2 mm. long or practically lacking; calyx 2-3 mm. long, the lobes subulate; corolla salverform, the tube 5-8 mm. long, markedly constricted at the mouth, the lobes 4-7 mm. long, ovate to oblong, spreading; stigma apiculate by two distinct obtuse lobes; follicles 6-8 cm. long, articulated into thickish constricted segments, sessile, conspicuously to- mentose, 3-7-seeded; seeds elliptic in outline, sharply tapering at both ends, and slightly arcuate, 12-13 mm. long, 3-4 mm. broad, reddish-brown. Distribution: mountain slopes and deserts, southern Nevada and southern California. Specimens examined: Nevapa: Eldorado Canon at Nelson, alt. 3000 ft., April 30, 1907, М. Е. Jones (Р). 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 423 CALIFORNIA: Cactus Ranch, Cushenberry Canon, San Ber- nardino Co., alt. 5500 ft., June 1, 1892, Parish 2412 (G); Mo- jave Desert, San Bernardino Co., June 5, 1915, Parish 10244 (G); San Bernardino Co., north slopes San Bernardino Mts., alt. 4000-6000 ft., June 15, 1895, Parish 3769 (G); Colorado Desert, in desert sands, alt. 3190 ft., 1921, Spencer 1778a (G); One-thousand Palms Canon, upper portion Colorado Desert, April 1, 1921, Jaeger 60 (Baker); Kelso, alt. 4000 ft., May 2, 1906, M. E. Jones (P, MBG); One-thousand Palms Canon, alt. 2700 ft., April 10, 1921, Jaeger 1173 (Baker); Keyes’ Ranch, alt. 3500 ft., common along wash, May 7, 1922, Munz & Johns- ton 5252 (Baker); vicinity of Corn Springs, Chuckwalla Mts. Colorado Desert, rocky slope in high gorge, alt. 2500 ft., April 9-12, 1922, Munz & Keck (Baker). 17. Amsonia arenaria Standley, Proc. Biol. Soc. Wash. 26: 118. 1913; Wooton & Standley, Contr. U. 8. Nat. Herb. 19: 505. 1915. Pl. 53, figs: 29-30. Herbaceous perennial from a thickened slightly woody root, tomentose; stems 2—4 dm. tall, clustered from the base, erect ог ascending, branched, the branches short, ascending; leaves nu- merous, crowded, alternate to subverticillate, sessile, linear- lanceolate to filiform, slightly fleshy, the midveins usually fur- rowed, 4-6 cm. long, 1-5 mm. wide; inflorescence dense, not usually held above the foliage; pedicels .5-3 mm. long or prac- tically lacking; calyx 4-7 mm. long, the lobes subulate; corolla salverform, the tube 8-10 mm. long, the lobes ovate-oblong to ovate-lanceolate, 5-8 mm. long, spreading; stigma apiculate by two distinct obtuse lobes; follicles glabrous, 5-8 cm. long, torose, articulated into 2-7 thickish constricted segments, sessile, glabrous, 2-7-seeded; seeds 1-1.5 cm. long, elliptic-arcuate in outline, sharply tapering at both ends, 3-4 mm. broad, nearly smooth, light brown. Distribution: gravelly plains and mountain slopes; Texas, New Mexico, Arizona, and Chihuahua. Specimens examined: UNITED STATES: Texas: exact locality lacking, 1857, Thurber 138 (Е). (Vor. 15 424 ANNALS OF THE MISSOURI BOTANICAL GARDEN New Mexico: San Andreas Mts., 1913, Wooton (G, US); exact locality lacking, 1852, Wright 1670 (G, MBG); Turney Range, Dona Ana Co., Sept. 23, 1912, Wooton (US); Strauss, rolling hills, 1912, Stearns (MBG). RIZONA: Cameron, infrequent, sand, June 7, 1922, Hanson 160 (F); same locality, along wash, June 8, 1922, Hanson 159 (MBG). Mexico: CHIHUAHUA: between Laguna de Guzman and Laguna Santa Maria, July 16, 1891, Hartmann 724 (G); gravelly plains near Lake Guzman, alt. 4000 ft., April 9, 1898, Pringle 6796 (G, P, MBG); among rocks, Ojo de Vaca to Los Plagas, June, 1851, Thurber 815 (G). Species EXCLUDED Amsonia orientalis Decne. in Jacquemont, Voy. Ind. 4: 105. 1841 = Rhazya orientalis (Decne.) A. DC. in DC. Prodr. 8: 386. 1844. List оғ ExsiccATAE The distribution numbers аге pue in italics. Тһе number in parentheses is the species number in this revision Albrecht, N. — (3). Allard, H. A. 197 (4a). Allen, T. F. — (4a). Anderson, E. S. & Woodson, R. E., Jr. 6 (4b); 4000 (2a). Arséne, Bro. — (2a). Bailey, V. 1885 (15). Baker, C. F. — (4b). Bartlett, H. H. 1493 (2a). Bartram, E. B. — (4a). Berg, N. K. — (2). Biltmore Herb. 81, 81b (4a). Blakely, G. 3425 (4 ). Blankinship, J. W. — (4a). Boyce, 8. 5. — (4a). Brandegee, K. — (14). Bray, W. L. 60, 239 (2). Brown, O. E. — (4). Buckley, S. B. — (2a); — Bush, B. F. 646 (2b); 11, T 2378, 7513 (4); 4461 (4a). Canby, W. M. 68 (2). Carpenter, — (4a). Chapman, A. W. — (1); — (2); — (2a). Clemens, Mrs. J. 11727 (2b). Commons, A. — (4). Cooper, C. H. — (4a). Coville, F. V. & Funston, F. 825 (14). Curtis, M. A. — (1); — (2); — (2a); (4a). Curtiss, A. H. 6820 (1); 2269, 6376 (2); 6476 (2a). Davis, J. 398, 1403 (4b). Donnell-Smith, J. — (2a); — (4). Douglass Drushel, 1. А. у (2); 8757 (48). (2). а). Eastwood, А. 72 (10); 73 (15). Edward, К. — (4 ) Eggert, Н. — (2); — (2b); — (4a). Emig, У. H. 899 (2b); — (4b) Engelmann, G. — (4); — (4b). 1928] WOODSON—STUDIES IN APOCYNACEAE. Ш 425 Evermann, B. W. — (14). Faxon, C. E. — (4a). (2a). Я : 48). т» 7. М. 3944 HA. 4498 (Ab). Hale, — (5). Hall, E. he E — (Ab). Hall, H. M. 6006 (14). Hanson, H. C. 159, 160 (17). Harper, В. M. 440, 448, 606 (1); 1819 (2): 577, 915, 1166, 2138 (2a). Harrison, G. J. & Peebles, R. H. 5540 (7). Hartmann, C. V. 724 (17). Harvey, F. L. 38 (4). H 3 азве, —. — (45) Havard, У. — (9 3). Hitchcock S. — (2a); 76a (4) Hopkins, M. H 3, Новду, Ј. В. — (да). Houghton, М. 3606 (2b). House, H. D. 1851 (4). Hubbard, G. W. 2268 (4a). Ingalls, T. J. — (5). Jaeger, E. C. — (14); 288 (14); 60, 1173 (16). Jermy, G. 145 (2b). Jones, M. Е. — (6); —, 5289aa, 5469, 5093, 54764, 6446 (10); — (14); — (15); — (16 Јоог, Ј. Е. — ( Ledman, О. 5. — (4b). Lemmon, J. G. 3248 (6); — (14). Lemmon, J. G. & Lemmon, — (6). Letterman, G. W. — (2a); — (4b). Ir F. — i" 660, 4 (2b). (4). . — (4а). Mearns, E. A. 152 (6); 117 (8). Meislahn, M. 210 (2a). Millspaugh, C. F. 4030 (4a). Mohr, C. — (2a Munz, P. A. 4535, 10505 (14). unz, P. А. & Johnston, I. M. 5253 (14); 5252 (16). Munz, P. A. & Keck, D. 4843 (14); — (16) Nash, G. V. 2546 (2a). Oberwetter, P. “ЖЕ (18). Olney, 5. Т. & Metcalfe, J. 76 (2). Painter, J. Н. — (4). Palmer, Е. 90 (12); 302, 435 (14). Palmer, Е. J. 27233 (2); 33031 (2a); 11471, 11616, 12233, 12262, 12597, 13496, 24203 (2b); 12, 818, 921, 4305, 8286 (4); 13090 (4а); —, 1819, 2620 (4Ь). Parish, 8. В. 1332, 2411, 3765 (14); 2412, #4 10244 (16 Parry, C. С. — (12); — (14). Peebles, к Н., Harrison, С. J. Kearney, Т. Н. 3820 (11); 6858 5). (1 Pitcher, — (4a). Plank, E. N. — (2); — (4b); — (13). Sis P$. ). rter, T. C. — (га); — (4b). Price, S. Е. — (4a). Pringle, C. G. 6796 (17). Purpus, C. A. 5988 (14). Ravenel, H. W. — (2 Rehn, J. A. G. & Viereck, H. L. — 12). Reverchon, J. 84 (2a); —, 698, 3122 (2b); 3881 (12); 99, 1557 (18). М. — (2). Rugel, Е. — (1); 21 (2a). Rusby, Н. Н. 256 (7). Ruth, А. 241 (2b); a 466, 480 (4). Sargent, С. 5. — (2); 69 (4a). Saunders, С. Е. — (14). Seymour, А. В. 1584 (4). Sheldon, С. 8. 224 (4b). Sherff, E. 801 (4b). Short, C. W. — (4b). [У ог. 15 426 ANNALS OF THE MISSOURI BOTANICAL GARDEN Siler, A. L. — (14). Smith, A. H. — (4a). Spencer, M. F. 1778, 347 (14); 1778a (16). Standley, P. C. — (4а). Stanfield, S. W. — (2). Stearns, E. 228 (9); — (17). Stevens, G. W. 29 (2b); 2670 (4); 2337 (4b). Sydone, O. — (2b). Taylor, K. A. — (2). ни Е. Н. 2018 (11). Thompson, Mrs. А. Р. — (14). Тћигђег, G. 188, 315 (17). Tuomey, M. — (4a). Vail, W. 546 (4). van der Velden, A. & W. — (2). Vasey, G. R. — (12 Ward, L. Е. — du); — (15) Weatherill, J. Eos Wells, E. — Wilkins, T. — "d Williams, E. F. — (4). Williamson, C. S. — (2a); — (4a). Woodson, R. E., — (4a). Woodson, R. Е, Jr. & Stevenson, J. 41 (4b). bis ec E. O. — ум, — (17). Wright, С. — (2a); 1669 (6); — (9); a m 72 (12); pe (13); 1670 iE M. В. — (2b). INDEX TO SPECIES New subgenera, sections, species, varieties, and combinations are printed in bold face type; synonyms in italics; and previously published names in ordinary type. Pag у & ‚ку, ут аса таа а па 395 т Опра он лалы а акта и ЭЭН 398 222” таг. TERIBOR уж да» 402 Т о v 423 brevifolia əға Ы 419 4. var. tomentosa......... 422 ШИНЭ ИЕ e be > 398 jen var. /Шойа............. 400 ciliata var. tenuifolia......... 400 ciliata var. texana.............. 402 Eastwoodiana................. 421 elliptica (Thunb.) В & S....... 403 "elliptica Sieb. & 2асс.”........ 403 Fremontü...... КЕТЕ ве В c. ira FS о ирэлт сс 413 ПН miro S RED 414 Kearneyana................. 415 кек ЗОО. ‚с cos keri viens 414 реда: MN ND eMe са. 404 то, Аи на 416 р ROO y^ 410 СЕТ СҰЛЫ ТТІ УГ 42 uo РИ 2 эл 411 pogonosepala an cn 412 ial ake chal ТМРА ee 397 Page SUING гд TENE ЭЭ 406 salicifolia var. ciliolata......... 400 salpignantha................ 417 SE Pee а она 413 Tabernaemontana............. 404 DM var. salicifolia 406 с ПИТЕР ere eer S 400 texana р РУС 402 Куф... 414 orc deo есь 422 ось TUM SS 405 Anonymus 2.” Атта mr 404 сул атан рақ ли 396 амалы. eee OU TIU 398 СИВ с рам 398 ПИ а 403 АОИ o 405 WONG, ЛАР ох PELA 407 MENS. (o и ње 400 ИШИ. Que мөрт” 418 о ЭН “ОР ОТТИ"? 396 1928] WOODSON—STUDIES IN APOCYNACEAE. III . 427 Page Page Епатвоп1а.................... 411 Tabernaemontana $ micranthae ПОНЕ: россии к 404 зарим јона 7. : „|. VER Pas C eR хэ 398 ШИН ое 424 rera г ао ока 403 [Уоь. 15, 1928] 428 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 50 Map of the geographical distribution of Amsonia in North America. ©000 subgenus Еиатзота (К. Schumann) Woodson. ..... А. ludoviciana. =a subgenus Sphinctosiphon (К. Schumann) Woodson. ^ meri. **** А. Standleyi. даа subgenus Articularia Woodson. —— А. brevifolia. еее“ А. Eastwoodiana. ННН А. arenaria. NOTE: Since the preparation of Plate 50, Amsonia ciliata var. tenuifolia has been found locally in extreme south-central Missouri (E. J. Palmer 33031; Anderson & Woodson 4000), thus extending the known distribution of that species from central Arkansas. 15, 1928 VoL. GARD. Bor. Mo. VN. Ме T mw at Зы Щр “зө, ща сал Ба Us «х, 7597 м д 211 ~ СЫ шэг “уы М M^ ~~ олай 4 ВИР n TIT с ПРАТИ JAN e Hel oF THE YNIreEeo STATES graphic Diagram Мас 2n, Wisconsin Mountains taken from A.K.Lobeck Physio ioo ит STUDIES IN APOCYNACEAE WOODSON [Vor. 15, 1928] 430 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 51 Illustrations of the subgenus Euamsonia, with comparison to Haplophyton; all figures Х 2 except when otherwise noted. Haplophyton cimicidium A. DC. Fig. 1. Flower. ХІ. Fig. 2. Front and side of stamen. Fig. 3. Pistil. А. rigida Shuttlew. Fig. 4. Flower. Fig. 5. Front and side of stamen. Fig. 6. Pistil. А. ciliata Walt. Fig. 7. Flower. Fig. 8. Pistil. A. elliptica (Thunb.) Roem. & Schult. Fig. 9. Flower. Fig. 10. Pistil. A. Tabernaemontana Walt. Fig. 13. ЁоШсїев and seed. X М. ANN. Мо. Вот. Garb., Vor. 15, 1928 PLATE 51 WOODSON—STUDIES IN APOCYNACEAE (У от, 15, 1928 432 ANNALS OF THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 52 Illustrations of the subgenus Sphinctosiphon; all figures Х 2 except when other- wise note A. Ратет Gray. Fig. 14. Flower. Fig. 15. Pistil. А. Standleyi Woodson. Fig. 16. Flower. Fig. 17. Pistil. A. longiflora Torr. Fig. 18. Flower. Fig. 19. Pistil. Fig. 20. ЕоШсіев and seed. X М. A. salpignantha Woodson. Fig. 21. Flower. Fig. 22. Pistil. ANN. Mo. Вот. Garb., Vor. 15, 1928 PLATE 52 WOODSON —STUDIES IN APOCYNACEAE [Vor. 15, 1928] 434 ANNALS ОЕ THE MISSOURI BOTANICAL GARDEN EXPLANATION OF PLATE PLATE 53 Illustrations of the subgenus Articularia; all figures х 2 except when otherwise ted. А. tomentosa Torr. & Frém. Fig. 25. Follicles and seed. х 14, A. brevifolia Gray. Fig. 26. Flower. Fig. 27. Pistil. Fig. 28. Follicles and seed. x 14. А. arenaria Standley. Fig. 29. Flower. Fig. 30. Pistil. 5 “~ ANN. Mo. Вот. Garb., Vor. 15, 1928 PLATE WOODSON —STUDIES IN APOCYNACEAE GENERAL INDEX ТО VOLUME ХУ New scientific names of plants PE ыг ца rasan айы» + new combinations are ока! in bold fac plates, in а type. ace type; synonym A Acanthaceae, A new genus of the, 1 Acetic acid, utilization of, by Azoto- bacter cultures 132, 139 ources of energy for Azo- Acids: rider. with specia dri: to, 113; volatile, recovery of, from stand- ard solutions, 120, u tilization со vari- ous, by Azotobacter altars Anderson, Edgar. The слей ет е зре- cies in the northern blue flags, Iris т L. and Iris virginica ТЕ „ 24 е genus, 379 0, а и о са, 403, 430; elliptica, 403; ы 411; hirt tella, 413; Jon nesii, arn eyana, 415; latifolia, 404, 6, 432 udoviciana, 2; tenuifolia, 400; texana. 402, 414; tomen- sa, 422 ; triste ies Ansonia. 396; ustifolia, 398; ciliata, 398; elliptica, 403: latifolia, 405; salici- folia ; tenuifoli Қолы T: а 2, 6, 8, 60; 1 15, 1, 6, 8, 60; айн ҚЫСЫН, 3, 6, 60 ж Studies in the, II, 341; шоо officinalis, 93 Articularia Ash weight ot plants, effect of ultra- violet о оп, 206 Athyriu ыы. Sources of energy for, with special reference to fatty acids, 113 Azotobacter chroococcum, 118; vine- landii, 118 ANN. Mo. Bor. Gard., Vor. 15, 1928 ving reference to figures italics; and previously Зара т диско and all other matter, B ИН А new genus of, 335 Bignonia obovata, 357 Bryophyllum, effect of ultra-violet radia- anatomy 9 Butyrie acid, utilization of, by Azoto- bacter cultures, 134, 140 C Calcium salt of fatty acids, ability of zotobacter € ш grow in, 149 Calophanes, пи Зен 54; angusti-. olius, 36; ri ET bila biatus, 39; bilobatus, 28: ; californ nica, 60; capita atus, 42; ciliatus, 48 ; crinitus ‚ 30; с ube nsis, 60; decumbens, 39; hirsutissimus, 2 hirsutus, 51; у бик AN аа, 3; hygrophi- loides, 51; Jasminum- мин icanum, bob lavandulace us, 32; linearis, 36; honis, 50; мира phils, MT; ML folia, 55, var. angusta 3l; NN folius ‚ 40, Ве texen ti 37; 0 ‚ 37, 53; Palm ‚ 60; pen Aca, “0; Ри- legium, 53: qua oua. 49; quiten- 818, 50; repens, 52; Schiedeanus, 34, var. ман aren US, 34; Serpyllum, 46: Twe edia 9 Саргоіс ud. utilization of, by Azoto- bacter culti А 144 astanopsis, Chlorophyll, effect of light rays on de- composition of, 204 — us, 109; cancellatus, 109; colum- diation on plants of, 180; anatomy of rayed leaves of, 198, of rayed stems, Colonnaria, 110 Corylus, 101 Cu ucumis, effect of ратом radiation natomy of rayed е пољ 201 о ч 5 E ха З бо © ~ 2 Cymopterus Ш ttoralis, 91, 93, 95, var. glaber, 93, var. glabr a, 99 Аан весі. Phellopterus, 92 (435) [Vor. 15 436 ANNALS OF THE MISSOURI BOTANICAL GARDEN D Glehnia, 91, 98, 101; эшти 95, 104, 108; littor alis, 93, 104, 106, 108: 2; humistr pon 3; linearis, 36; oblongi- olius, 55; riparius, Dyschoriste, А monograph x the Ameri- si diffusa, 1, 60; M к=. e^ 1 ода anii, 3 74; — й 1 lavandulacea, anhonis, 50, ые ек) 9, Шо phylla, 47, 86; Niederleinii, 34; о censis, 43, 78; a үүлэн ‘55, d f. glabra, 57; ovata ; paraguari n ana, 34; Schottiana, 30; Ser pyllum, 46; trichanthera, 29, 66; ава, 59: xylopoda, 54, 88 ysosma; а new genus of Berberidaceae, 335 Dysosma, 338; pleiantha, 339, 340 E Боож "тете Eltinge, Ethel The p of ultra- vio и: EL upon higher plants, 169 Euamsonia, 396 $ Еиатзотла, 396 Eustemmadenia, 353 Flags, "Erud blue, Problem of species in the, Formic Ан utilization of, by Azoto- bacter cultures, 132, 139 Gainey, P. L. Sources of en ал zotobacter, with special аА 10 fatty acids, § Galleottiae, 359 Genetic relationships of мээн dif- ferences in 1118 versicolor, 2 littoralis, 94, H Haplophyton, 387; cimicidium, 430 Hybridization of Iris versicolor and Iris virginica, Hydrogen-ion мэн of plants, effect of ultra-violet rays on, 204 Hygrophila brasiliensis, 60; “ааа 80 1 Ipomoea, effect of ultra-violet radiation on plants of, 180; anatomy of rayed leaf of, 196 Iris, 241; americana versicolor, etc., 254; carolina, 248, 254, 316; caroli їтїата, 248, 254; corollis imberbibus, 247, 254; foetidissima, ; fulva, 286; georgi- na, 2 4; hexagona, 296; latifolia Vir- d 547, 254, 314; prismatica, 286; ХЕ 26; Shrevei, 248, 254; versicolor, 247, 253, 316, 320, 324 326, 0; versicolor, 254, var. vir Iris е L. and Iris virginica L., em of шинэ ш те acid, utilization of, by Azo- tobacter cultures, 134, 141 J Jordanons, 307 Juglans, 101 K Kobuski, C. чк” А new genus of the Acanthacea e, 1; A monograph of the уоона emg pate Е. of the genus Dys- choriste L хийц, жі of рас radiation n plants of, 181; anatomy of rayed wit ves o T 99 Т.агвеп, Esther 1. A new variety of Vernonia ин 333 Laternea, Concerning the status of the enus, Laternea, 110; angolensis, 110; bicolum ta, 110; columnata, 110; pusilla, 110; и. 110; Spegazzini, 110: всара, 110, 112 Linder, D. H. Concerning the status of the genus Laternea, 109 Linostylis, 2 § longiflorae, 416 192$] INDEX M poris. К айаш me 71818, « о Mathias, Mildred Е. и mbelliferae, I, 91 $ Micranthae, 411 N Nicotiana, effect of ultra-violet radia- tion при ts of, 180; а мэн my of of, 197, of sni ge s, 202 21:77 сена. 113; ейес о! pend pn 371; pana- Studies in the Меса total, recovery of, from Azo- tobacter cultures, 119 O $ obovatae, 353 с aphne, 363 Odontostigma, 352; Galeottiana, 361 Р Pasania, 101 Phaseolus, effect of ultra-violet radia tion on ша Xf, 182, 187, $e өлді» omy of ra leaves of, 200, of rayed ste | 08 Phellopterus, 92 хэсэн нийн, 91, 92, littoralis, 94, 95, 96 кр, 335; digforme, 339; 'Emodi, 335, 340; Esquiro ji, 339; Онгог, 339; Шо Henin, 339; versi- ems, Problem of мане п the northern рен flags, Iris лете a L. and Jris , 241 а; acid, utilization of, by Azoto- bacter cultures, 132, 139 R Raphanus, effect of Кати radia- tion pi of, 181; anatomy of s of, 2 Rhazya ог ааа Ruellia, 10; biflora, #55; саШогшса, 60; candida, ; cernua, 10; ciliata. 48; depressa, 10; diffusa я дет iniflora г. humilis, 58; humistrata, 3 i- Эн ры 5) litoralis, њи oblo ngifolia, 10, 55; ova a, 37, 53; eninsularis, 60; qui itensis, 507 repens, ЁЛ viscosa, 50 а «4 ЈИ 5 5рш inctosiphon, 4 $ Sphinctosiphon, 41 | 4 18 437 Spigelia scabrella, 6 Star ite “ч ай, in pants effect of ultra- et га Stevan eae prs * Аман, 360: bella, 861, ; bignoniaeflora, m сай уста, 358; ^ : nell-Smithii, 369; , eubracteata, a 378; Galeot- tian жим, ‚КО Дан га, 356, 374; мила эй 364 74; Greenmanii, 360, 376; рез. Н 371; insignis, 361; rg аи 362; mollis 358 obovata, 357, var. molli сха 0: sinaloans, ntosa, з ; Stemmadenia, ka vid of the genus, Stemmaderia, 352; Galeottianum, 361 T Tabernaemontana Alfari 360; Amsonia, 369; angustifolia, 398; Donnell-Smithii, 369, var. c | is 9; 9 га, 364; humilis, 404; laurifolia, 361 ‘Transpir ation, effect of ultra-violet ra- diation upon, 207 U Ultra-violet radiation, Бэхэн of, upon higher plants, 169; on lower organ- isms, 170; x най х Хүлгийн та 5, 175; enetration of, 175 Чайна, Studies in the, I, 91 V Valeric acid, utilization of, by Azoto- bacter cultures, 135, 141 Variation, intra specific, in Iris versi- _ colo or an 58 meri, 333; var. pir a 333, py W Weight, dry, of plants, effect of ultra- violet radiation ж, 205 W С Dysosma: а пе genus of њи еае, 337; Studies in the Apocynaceae, lI, 341; ПТ, 379 Z Zahlbrucknera mar — is, 50 Zea Mays, effect o ет м»: tion on " plants 185; a my rayed leaf of, 198, of rayed үгэн 202 leu- ^ s ї На Е Ж $ L5 ; 2 «x ( Циг foe Volume XV dea 171 Annals Missouri Botanical | Garden | Contents i February, 192 A New беа. of the Acanthaceae... Ron ; А EE .. Clarence E. Kobuski А Monograph of the American Species of the бейше Dyschoriste., аи, “17417224 апре Е. Kobuski Studies in the Umbelliferae. 1...... го. Mildred E. Mathias Concerning the Status of the Genus ain | ‚ David Н. Linder PUBLISHED QUARTERLY AT 8 WEST KING наат, LANCASTER, РА., BY Me OF TRUSTEES OF THE MISSOURI BOTANICAL GARDEN, ST. LOUIS, ‘MISSOURI. Ra = matter at the post-office Ж отур Pennsylvania, under the Act of March 3, Number 1 Annals : of the Missouri Botanical Garden | А Quarterly Journal containing Везе бо И from the Missouri Botanical Garden and the Graduate Labora- tory of the Henry Shaw’School of Botany of Washington Uni- versity in affiliation with the Missouri Botanical pariete. 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