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CONTENTS OF REEL 243 1) Contributions from the Botanical Laboratory, vol. 9 MNS# PSt SNPaAg243.1 2) Contributions from the Botanical Laboratory, vol. 10 IVINS# PSt SNPaAg243.2 3) Contributions from the Botanical Laboratory and the Morris Arboretum of the University of Pennsylvania, vol. 11 MNS# PSt SNPaAg243.3 4) Contributions from the Botanical Laboratory and the IVIorris Arboretum of the University of Pennsylvania, vol. 12 MNS# PSt SNPaAg243.4 Title: Contributions from the Botanical Laboratory, vol. 9 Place of Publication: Philadelphia Copyright Date: 1932 Master Negative Storage Number: MNS# PSt SNPaAg243.1 <2252643> *OCLC* Form:serial 2 lnput:HHS EditFMD 008 ENT: 990118 TYP: d DT1: 1892 DT2: 1932 FRE: z LAN: eng 035 (OCoLC)7844150 037 PSt SNPaAg241 . 1 -243.2 $bPreservation Office, The Pennsylvania State University, Pattee Library, University Park, PA 16802-1805 090 10 580.8 $bP3c $cax $s+U1X1892-U10X1932 090 20 Microfilm D344 reel 241 .1-243.2 $cmc+(service copy, print master, archival master) $s+U1X1892-U10X1932 245 00 Contributions from the Botanical Laboratory 246 1 $iVol. 2 (1904)- have title: $aContributions from the Botanical Laboratory of the University of Pennsylvania 260 Philadelphia $bUniversity of Pennsylvania Press $c1 892- 300 V. $c25 cm. 310 Irregular 362 0 Vol. 1,no. 1- 362 1 Ceased with v. 10 in 1932. 500 Title from cover. 533 Miaofilm $mv. 1 -v. 1 0 $bUniversity Park, Pa. : $cPennsylvania State University $d1999. $e3 microfilm reels ; 35 mm. $f(USAIN state and local literature preservation project. Pennsylvania) $f(Pennsylvania 533 Microfilm $mv. 1 -v. 1 0 $bUniversity Park, Pa. : $cPennsylvania State University $d1999. $e3 microfilm reels ; 35 mm. $f(USAIN state and local literature preservation project. Pennsylvania) $f(Pennsylvania agricultural literature on microfilm). 650 0 Botany. 650 0 Botany $xBibliography. 710 2 University of Pennsylvania. $bBotanical Laboratory. 785 00 $tContributions from the Botanical Laboratory and the Morris Arboretum of the University of Pennsylvania 830 0 USAIN state and local literature preservation project. $pPennsylvania. 830 0 Pennsylvania agricultural literature on microfilm. Microfilmed By: Challenge Industries 402 E. State St P.O. Box 599 Ithaca NY 14851-0599 phone (607)272-8990 fax (607)277-7865 www .htm IMAGE EVALUATION TEST TARGET (QA-3) 1.0 I.I 1.25 1m msA I&3 mi ■ao 2.8 3.2 i ii 3.6 4,0 1.4 2.5 2.2 2.0 1.8 1.6 150mm // /APPLIED A IM/IGE , Inc -^= 1653East Main street ^:: Rochester, NY 14609 USA := Phone: 716/482-0300 Fax: 716/288-5989 © 1993, Applied Image, Inc., All Rights Reserved CONTRIBUTIONS FROM THE Botanical Laboratory OF THE UNIVERSITY OF PENNSYLVANIA VOLUME IX 1931-1932 PHILADELPHli •fc'Cn.' ;i 'C^ 1 1> 3 2 • • « • • •'•• •» ». », • « • •• • • • • • •• v* • • •• • • • • • •• ' • ' • • • • • • # • • • •• « • •.'^^ •' >« • • •••• j^, ••• CONTENTS •' > < • • • • • • < « ^ , •^ • • « • • • • • • • * • • • • • .«.'» ' * • • •!•''• »' •« •• • '! • •• CLARK, RUTH S. Stem anatomy of tomato, Lycopersicum escu- lentum L. (Proc. Pennsylvania Acad. Sci. 6:1-3. 1932.) FOGG, JOHN M. Notes on a few introduced species in Philadel- phia local area. (Bartonia 13:48-49. 1931.) RICHARDS, HORACE G. Notes on the marine algae of New Tersev (Bartonia 13:38-46. 1931.) SEIFRIZ, WILLIAM. Radiant energy from living matter. (Sci. Education 16:34-37. 1931.) 1 Sketches of the vegetation of some Southern provinces of Soviet Russia. I. The altitudinal distribution of plants on the Crimean Mountains. II. Plant life along the Georgian military way. North Caucasus. (Jour. Ecology 19:360-382. 1931.) TRUE, RODNEY H. The introduction of Lobelia syphilitica into medicine. (Amer. Jour. Pharm. April, 1932.) John Bartram's life and botanical explorations. (Bartonia 12 (Suppl.):7-19. 1931.) -Kyllinga pumila in Philadelphia. (Bartonia 13:47. 1931.) TUAN, HSU-CHUAN. Unusual aspects of meiotic and post-meiotic chromosomes of gasteria. (Bot. Gaz. 92:45-65. 1931.) WHERRY, EDGAR T. The Eastern long-styled Phloxes, part I. (Bartonia 13:18-37. 1931.) The Eastern long-styled Phloxes, part II. (Bartonia 14:14-26. 1932.) Ecological studies of serpentine-barren plants — I. Ash composition. (Proc. Pennsylvania Acad. Sci. 6:32-38. 1932.) -Range-extensions and observations, 1931-32. (Amer. Fern Jour. 22:79-86. 1932.) ZIRKLE, CONWAY. Some forgotten records of hybridization and sex in plants. (Jour. Heredity 23:432-448. 1932.) ^Vacuoles in primary meristems. (Zeitschr. Wiss. Biol. Abt. B. Zeitschr. Zellforsch. u. Mikroskop. Anat. 16(l):26-47. 1932.) 145024 00 do Reprinted from Proceedings op the Pennsylvania Academy op Science Vol. VI, 1932 STEM ANATOMY OF TOMATO, LYCOPERSICUM ESCULENTUM L/ Ruth S. Clark Department of Botany, University of Pennsylvania The tomato has so recently attained its wide-spread popularity as an esculent that it is not surprising its anatomy has not formerly been studied. The group to which the tomato belongs boasts several economi- cally important food plants whose anatomy has only lately been studied, such as the potato in 1917, and the eggplant in 1931. It was particu- larly to note the similarity, if any, to these related plants that the pres- ent study was undertaken, with special reference to the origin and devel- opment of the internal phloem. The fact that the presence of internal phloem has been noted in many families does not make it less easy to discover its origin. It has its be- ginning in the tomato in the rather long transition area, for while seen clearly developed in the mature stem, it most certainly is not to be seen in the diarched stele of the root. Therefore by taking successive trans- verse sections of the transition area (hypocotyl) we shall be able to note the progress and development from root to stem. Let us examine a section at the ground line. Here the elements are arranged in diarched manner typically root-like. The protoxylem ele- ments may be slightly out of the straight line and the phloem areas may be showing signs of dividing, but the general aspect is close to root structure. At approximately mid-height of the hypocotyl, which is from one and one-half to two inches in length, great changes can be noted. The four bundles are practically if not entirely separated, each with its own ex- ternal patch of phloem. Then somewhere between these converging xylem bundles are seen two (splitting) or four discrete areas of phloem — destined to become the internal phloem. As yet they are not fully orientated to their position on the inner edge of the xylem elements. 1 This paper is part of a thesis presented to the Faculty of the Graduate School of the University of Pennsylvania in partial fulfilment of the requirements for the degree of Master of Science. Further details and illustrations can be noted by refer- ence to the thesis entitled * * Morphology and Anatomy of the Tomato Plant ' ' (Lyco- persicum esculentum L.) With Special Reference to the Internal Phloem. 1/500 Just under the cotyledonary outgrowths a transverse section shows the elements characteristically arranged as for the mature stem, the ex- ternal phloem, cambium, xylem, and internal phloem in order, outermost inward. May I recall rather briefly some of the theories which have been advanced for the existence of internal phloem. Some writers believe that it is the vestigal trace of bundles which were formally complete. Other authors consider that this tissue has been built up because of the need of additional conduction tissue. This latter view has been adopted by Artschwager as the case noted in the growth of the potato — this work will be noted below. DeBary, one of the earliest workers, believed the internal phloem was connected to the external phloem by a narrow band of sieve tubes. These were very tiny cells and often escaped notice ; however their pres- ence causes the so-called ''bicollaterar' bundles to be, in reality, con- centric. AYorsdell is the advocate of the vestigal idea of internal phloem. He says the now existing plants showing bicollateral bundles originally pos- sessed a scattered bundle area such as we typify in the monocotyledon- ous arrangement. Of this scattered system two series remain — the out- ermost is complete, the phloem of the second is the only remnant of that inner ring of bundles. In the work done by Scott and Brebner they see that during the tran- sition period successive strands of phloem split off and move outward with the converging of the xylem elements, and, fusing, form the phloem area. These authors consider the internal phloem as an asset because of its increased protection, being inside the wood cylinder, and its near- ness to the pith facilitating communication with this latter tissue. While working with the Cucurbitaceae, Holroyd said that cambioid cells develop very shortly after the first splitting of the xylem in the hypocotyl enclosing the bundle, so that it might be called a *'perixy- lary" cambial ring. This cambium gives rise to the internal phloem, but continues to remain dormant on the sides of the bundles. Now according to Artschwager the phloem initials are first to be noted internally, the external phloem develops at a latter period. This succession, however, is probably true only for the potato. In my work on the Lycopersicum esculentum, or tomato, I am in- clined to think the facts noted are most in agreement with those set forth by Scott and Brebner. There is merely a replacing of part of the first phloem areas — no secondary initials or cambial action being involved. Strands of the external phloem move inward during the transition period and become orientated at the center or base of the xylem ele- ments. Division of the phloem cells themselves accounts for the addi- tional internal phloem, while the splitting of areas continues the '* broken ring'' arrangement for this tissue. At no time could evidence of any cambium be seen. Reprinted from Bartonia, No. 13, 1931. Notes on a Few Introduced Species in the Philadelphia Local Area John M. Fogg, Jr. Artemisia biennis. Among the plants frequently collected on ballast ground near Philadelphia 40 or 50 years ago was the biennial wormwood, Artemisia Uennis Willd. This spe- cies, the natural range of which is given in Gray's Manual as Ohio to Tennessee, Missouri, and northwestward, was long known as an introduction at Kaighn 's Point, Camden County, New Jersey, and at several localities in Philadelphia County on the Pennsylvania side of the Delaware River, notably from below the Old Navy Yard. One of the last collections of this plant from Camden County was made by Albrecht Jahn, who, in 1898, found it still growing at Kaighn 's Point. Since that date, however, there appear to be no records of the plant's occurrence on the New Jersey side of the river, although it has been twice collected in Philadelphia County ; having been found by C. S. Williamson at Greenwich Point in 1911 and by Dr. H. B. Meredith at the foot of Wolf Street in 1920. Its recent appearance in a freight yard at Cooper's Point, Cam- den County, is, therefore, a matter of some interest. The plant was here discovered by William H. Witte of Camden, who reports that it was growing with two other species of the genus, A. annua L. and A. vulgaris L. Specimens of all three species have been deposited in the herbaria of the Philadel- phia Club and the University of Pennsylvania. CoRONOPUS DiDYMUS. Another plant formerly of frequent occurrence on ballast ground and in waste places, but not well represented by recent collections, is the wart cress, Coronopus didymus (L.) Sm. Material in the herbarium of the Phila- delphia Botanical Club show that within the last 30 years this species has been found at only two localities in the local area. H. L. Fisher collected it at Hatboro, Pa., in 1905, and Harold W. Pretz found it growing as a weed in the pavement at Allentown, Pa., in June, 1930 (Pretz No. 13341). It is en- tirely possible that Coronopus is not so rare as these scattered (48) PHILADELPHIA BOTANICAL CLUB 49 collections seem to indicate, since the plant is by no means a conspicuous one and might readily be overlooked by collectors. Nevertheless, in view of the marked tendency of the species to become a pernicious weed in lawns, it seems worth while to place on record a third locality for its recent occurrence. This interesting little Crucif er made its appearance on the campus of the University of Pennsylvania in 1924 on the lawn in front of Zoological Laboratory. Since that time it has spread rapidly, occupying the ground bordering Hamilton walk for a distance of about two city blocks, and has so far resisted all efforts to eradicate it. Cardamine pratensis. Although very abundant in regions to the northeast, the bitter meadow cress or cuckoo flower, Cardamine pratensis L., is a rare plant within the Philadel- phia local area. Material in the collections of the Philadel- phia Botanical Club show that it was collected as a weed in a lawn in Annandale, Hunterdon County, New Jersey, by H. L. Fisher in 1905. Twenty-one years later Mr. Fisher again collected the plant in Annandale, this time **in an old yard," and his two sheets in the Club Herbarium dated 1926 indi- cate, if they be from the same locality as the 1905 collection, the ability of the species to persist when once established. In the Spring of 1924 Harold W. Pretz collected the plant near Laury's Station in Lehigh County, Pennsylvania, where it was found growing in a ** wooded, marshy place," a habitat more nearly like that adopted by the species in New England and Nova Scotia. It is now possible to add a third locality to these two, as it was the writer's good fortune to find this at- tractive plant in low-lying wet ground along Little Tacony Creek near City Line, Oak Lane, Philadelphia County, Pa. This station was first discovered in May, 1924, and observa- tions covering several years showed that the plant was spread- ing. The locality appears doomed to extinction, however, for the waters of the creek have been diverted and the ground is rapidly being filled in by artificial means. Specimens from the original collection (Fogg No. 648) have been deposited in the Philadelphia Botanical Club and the University of Penn- sylvania herbaria. University of Pennsylvania. Beprinted from Babtonia, No. 13, 1931. Notes on the Marine Algae of New Jersey^ Horace 6. Eichabds During the past few years the writer has been engaged in a survey of the marine invertebrate animals of the coast of New Jersey, with especial reference to the Cape May region (Richards, 1929). During the course of the collecting some attention has been given to the Algae, and it is here intended to record the most conspicuous marine plants of the region. The Algae of the state have been collected and listed by numerous workers, but nothing about them has appeared in the past twenty years. One who compares the present list with that of Martindale or Collins cannot fail to note that it is much smaller. This is undoubtedly due in part to insufficient collecting, but it is also thought that various fac- tors such as the pollution of the water by oil, sewage, dye- wastes, etc., the draining of the marshes, the development of the land, and marine erosion, have played some part in the depletion of the Algae of the region. Within the memory of the writer a great many more species could have been collected at Longport, N. J., than may be found there at the present time. As a more specific example in the reduction of the marine flora of the region, we could mention the case of upper Cape May Harbor near Schelen- gers Landing. During the fall of 1928 a new bridge was built at this point. In the progress of the construction considerable oil was poured into the water. The Algae and many of the marine invertebrates on the dock near-by were soon killed, and even by the following summer when the work had been com- pleted, they had not entirely regained their foothold. Description of the region. The shore-line of New Jersey from Bay Head to Cape May is made up of coastal islands, separated from each other and from the mainland by bays and 1 Contribution from the Botanical Laboratory of the University of Pennsylvania. Published with the permission of the Secretary of the Smithsonian Institution. (38) PHILADELPHIA BOTANICAL CLUB 39 inlets. These islands vary in width from a few hundred yards to more than a mile. Above Bay Head there are numerous inlets but the mainland extends down to the ocean. On the bay side of these islands is the salt marsh. The extent of this has been greatly diminished by the very commendable work of the state of New Jersey in attempting to exterminate the mosquito. Eel grass {Zostera marina L.) is present in patches in Cape May Harbor and the other inlets of Cape May County, but is not as abundant as it is in Barnegat Bay and the sounds north of Atlantic City. A rock jetty, a mile long, has been built at the entrance of Cape May Harbor (Cold Spring Inlet). This affords a foot- hold for animals and plants which might not otherwise be found in the region. Previous work on New Jersey Algae. The first important work on the marine Algae of the eastern coast of North Amer- ica is the classical work of Harvey (1852). He divided the flora of this region into four zones, of which one was *'Long Island Sound, including under this head New York Harbor and the sands of New Jersey.'' Shortly afterwards Mr. Samuel Ashmead (1857) published a catalogue of the marine Algae discovered at Beesleys Point, Cape May County, during the summer of 1855. He recorded five browns, nineteen reds and five greens. Farlow (1881) in his work on the Algae of New England states that little had been recorded from the shores of New Jersey, and expresses the belief that the sandy shores of the region would support a very poor flora of marine Algae. Collins (1888) recorded one hundred and seven species of Algae collected by Mr. S. R. Morse at Atlantic City, a number much greater than could possibly be found at this place at the present time. In 1889 Dr. N. L. Britton edited a catalogue of the plants found in New Jersey. The list of marine forms was contrib- uted by Isaac C. Martindale and includes records from all sections of the coast. This is the most complete list of the marine Algae of New Jersey that has ever appeared. 40 PROCEEDINGS OF THE A brief description of some marine Algae found at Long- port, N. J., was given by Mrs. M. S. MeCullough in the ** Daily Union History of Atlantic City and County" (1900). Methods. During the early part of the investigation (1928-29) the Algae were obtained by shore collecting or from a row boat in the various inlets and thorofares in the region. During the summer of 1929 and the early months of 1930, through the kindness of Dr. A. E. Parr of the Bingham Oceanographic Foundation (New Haven, Conn.) and by ar- rangements with the United States Bureau of Fisheries, the writer was able to make numerous dredgings in Delaware Bay and at a few stations in the ocean off Cape May. In 1931 the work of the Bureau was expanded to include both inshore and offshore waters between Cape May and upper Barnegat Bay. The presence of a shifting sandy bottom off this coast does not favor the growth of Algae, and therefore it is not surpris- ing that very few species were dredged offshore. The inland waterways, however, were found to be rather rich in Algae, although in individuals rather than in species. From the ob- servations of May to August, 1931, the whole inland water- way from Cape May to Barnegat Bay might be termed a sin- gle association with Viva lactuca and Agardhiella tenera the dominant species. The writer is indebted to Dr. William R. Taylor of the Uni- versity of Michigan and to Dr. John M. Fogg, Jr., of the Uni- versity of Pennsylvania for valuable help in the identification of specimens, and for suggestions and encouragement in the course of the collecting. Sets of specimens are being deposited in the United States National Herbarium and in the herbarium of the University of Pennsylvania. Annotated List of the Marine Algae of New Jersey Myxophyceae (Blue-green Algae) Species of Lynghya, Phormidium, Spirtdina and Oscilla- toria have been found on piles throughout the region. The PHILADELPHIA BOTANICAL CLUB 41 blue-green Algae are usually found near high water mark, or even slightly above. On piles they are frequently above a zone of Enteromorpha, Chlorophyceae (Green Algae) Enteramorpha intestinalis (L.) Link. Common with E. campressa on rocks and piles throughout the year; usually between tides ; dredged in shallow water in the inland water- ways and occasionally in the open ocean. E. compressa (L.) Grev. Common with the above between tides. E. lima (L.) J. Agardh. On shells, etc., in the inlets; especially in spring and fall. E. plumosa Kiitz. This and other small species of Entero- morpha have been found attached to eel grass in the inlets. E, clathrata (Roth) Grev. Cape May Harbor, Great Egg River, Forked River. Viva lactuca (L.) LeJolis. (Sea Lettuce). Very abundant in all inlets and thorofares in the region, even in very brack- ish water; dredged in Delaware Bay and the ocean several miles off New Jersey. Cladophora spp. Various species are common in the region ; wharves, mud flats, ditches, etc. ; a specimen dredged in Great Sound (Stone Harbor) on August 12, 1931, in 9 feet of water, according to Dr. Taylor *' seems to be C hrachyclona Mont, or C. utriculata Kiitz., French rather than Caribbean form." Bhizoclonium riparium (Roth) Harvey. Cape May Har- bor, April 7, 1928. Chaetomorpha linum Farlow. Dredged in Corsons Inlet and Ludlam Bay in summer of 1931. Bryopsis plumosa (Huds.) C. Agardh. Rare; attached to wharf at Cape May in September and October, 1928 ; absent from same locality in 1929 due to oil pollution. Hormiscia pennicilliformis (Roth) Fries. On Rock Pile, Cape May, March 11, 1928. Vlothrix flacca (Dillw.) Thur. (sp.?). On rocks between tides. 42 PROCEEDINGS OF THE Phaeophyceae (Brown Algae) Ectacarpus sUiculosus (Dillw.) Lyngb. Occasionally found in inlets, from May to October. E, confervoides (Roth) LeJolis. More common than the above; especially in cold weather (December and January.) Ascocylus orbicularis (J. Agardh.) Magn. {Myrionema orUcularis Agardh.). On floating Zostera, December 2, 1928. PylaielU littoralis (L.) Kjellm. On wood-work; common in late spring and summer. Elachisia fucicola (Veil.) Fries. On Fucus, in late sum- mer and fall. Punctaria plantaginea (Roth) Grev. On beach at Cape May in winter, after storm of January 27, 1929; Beach Haven, February 22, 1931. P. latifolia Grev. Fairly common in harbor in spring and early summer. Arthrocladia vUlosa Duby. Abundant at various stations off Wildwood and Cape May, especially at Five Fathom Bank, 14 miles off Cape May, during the summer of 1931. Was not found at these localities during the summers of 1929 and 1930. Dictyosiphon foeniculatus Grev. var. americanus Collins. Cape May Harbor, May 20, 1928. Scytosiphon lomentarius (Lyngb.) J. Agardh. Rather abundant in the Harbor in the spring ; taken in considerable numbers on April 7, 1928 ; Beach Haven, February 22, 1931. Laminaria agardhii Kjellm. (Devil's Apron). Occasion- ally found washed on the beach near Cape May ; common on the rock jetty at Atlantic Highlands, N. J., on August 1, 1928; Beach Haven, February 22, 1931; dredged, Barne- gat Bay, Forked River, July 7, 1931, in 1 fathom. Fishermen report that on various occasions they have encountered a dense growth of Laminaria at Five Fathom Bank, 14 miles off Cape May (5-9 fathoms). No specimens of this alga were dredged at this locality between 1929 and 1931. Fucus vesiculosus L. (Rock Weed, Crab Grass, Poppers). Very common on rocks and piles near low water mark throughout the year. PHILADELPHIA BOTANICAL CLUB 43 Sargassum filipendulum C. Agardh. (Gulf Weed). Washed on the beach September 26, 1929. Ascophyllum nodosum (L.) LeJolis. Beach, Cape May Point, October, 1929; dredged Great Sound (Stone Harbor, July, 1931). Rhodophyceae (Red Algae) Bangi. elegans (Mart.) Ag.) Dredged in large quantities in Barnegat Bay near Lavalette and in Toms River in July 1931 (1 fath.). The water here was practically fresh and the alga was growing in association with Zannichellm palustris. No trace of Dasya was found in the same localities in August. The only other record of this species is from Great Egg River near Jeffries Landing (also considerably brackish) on July 3, 1931, in 5 fath. Amphibia rivularis (Harvey) Kuntze. {Bostrychia rivu- laris Harv.). Found on piles below low tide where the water is slightly brackish; collected on roots of Salicornia sp. in the back channel at Avalon (Aug. 28, 1928); present throughout the summer and fall. Also collected at Cape Charles, Va. (June, 1928). Chondria laUeyana (Mont.) Farlow. Dredged in Great Bay and Little Egg Inlet, July 6, 1931. Polysiphonia variegata (C. Ag.) Zanard. Common in the region throughout the summer and early fall; dredged in abundance at practically all inshore stations between Cape May and Barnegat Bay; occasionally dredged in Delaware Bay and in ocean off Barnegat, Atlantic City and Cape May. P. violacea (Roth) Grev. Rare; Little Egg Inlet, July 6, 1931 (5 fath.) P. nigrescens (Dillw.) Grev. Rare; occasionally found on beach after heavy seas; dredged Little Egg Inlet (4 fath.), Aug. 14, 1931 ; Great Sound, Aug. 12, 1931 ; Barnegat Bay, Aug. 15, 1931. P. olneyi Harvey (?). Rare; dredged near McCrie Shoal, 7 miles off Cape May (9i fath.), July 23, 1931. PHILADELPHIA BOTANICAL CLUB 45 P. harveyi Bailey. Rare; Barnegat Bay off Waretown, Aug. 15, 1931. Spyridia filament osa (Wulf.) Harvey. Dredged in Dela- ware Bay off Lewes, Del. in 5 fath, Aug. 21, 1929. Seirospora griffithsiana Harvey (sp. ?). On Zostera near Rock Pile, Cape May, Sept. 19, 1929. Antithamnion cruciatum (C. Ag.) Ag. Nag. On wharves at Cape May in September associated with Callithamnion roseum and Polysiphonia variegata. CMithamnion hyssoideum Arn. Not common; floating in the harbor on July 12, 1928. C. roseum (Roth) Harvey. Rather common on wharves at Cape May and Wildwood in September and October; not present in early summer. C corymhosum (Eng. Bot.) C. Ag. Attached to Zostera in harbor, December 31, 1928. C. haileyi Harvey. Rare; found on only one occasion on beach at Cape May Point after a severe northeast storm (August 13, 1928). Ceramium ruhrum (Huds.) C. Ag. Very common; found at all seasons of the year attached to eel grass in the inlets or attached to wharves. During the summer of 1931 it was dredged in inland waterways between Cape May and Barne- gat Bay, in Delaware Bay, Great Egg, Mullica and Forked Rivers and in ocean off Barnegat, Beach Haven, Brigantine and Cape May (5 to 7 fathoms). C. striatum (Kiitz) Howe. Quite common attached to Zostera on wharves during summer and early fall; dredged in inland waterways at Cape May, Great Bay, Longport, Little Egg Inlet; off Brigantine (5 fath.), Delaware Bay near New England Creek (2 fath.) C. tenuissimum (Lyngb.) J. Ag. With the above on wharves in summer and early fall ; dredged Great Egg River and Bay, Townsend Inlet, Barnegat Bay and off Brigantine Inlet (5 fath.) Agardhiella tenera (J. Ag.) Schmitz. The most abundant red alga in the inland waterways; dredged at every station 46 PROCEEDINGS OF THE between Cape May and Toms River; also collected in Dela- ware Bay and in ocean off Cape May. Present in Cape May Harbor in winter (Dec. 2, 1928, and Jan. 27, 1929). Melohesia lejolisii Rosan. Frequently found on floating Zostera in the harbor in summer and winter. M. pustulata Lamour. Not as common as the above; on floating Zostera in the harbor, Jan. 27, 1928. BIBLIOGRAPHY AsHMEAD, Samuel 1857. * * A Catalogue of Marine Algae discovered at Beesley 's Point during the summer of 1855.'' Geol. County of Cape May. pp. 152-154. Collins, F. S. 1888. * * Algae from Atlantic City, N. J. " BuU. Torrey Bot. Club 15: 309-314. Farlow, W. G. 1881. **The Marine Algae of New England and adjacent states." Eep. U. S. Fish. Comm. (1881). Haevey, W. H. 1852. ** Nereis Boreali- Americana. " Smithsonian Contribution to Knowledge. Maetindale, Isaac 1889. Algae in ''Catalogue of Plants found in New Jersey" (N. L. Britton). Geol. Surv. N. J. Final Rept. State Geol. 21; 384-467. McCullough, M. S. 1900. Marine Algae in *'The Daily Union History of Atlantic City and County.'' Richards, Horace G. 1929. *'A Faunistic Survey of the marine invertebrates of New Jersey, with especial reference to the Cape May Re- gion." (pp. 1-114.) Manuscript — to be published. Oct. 1931] RADIANT ENERGY FROM LIVING MATTER 35 M Radiant Energy From Living Matter William Seifriz* Department of Botany, University oj Pennsylvania, Philadelphia, Pa, Eight years ago the Russian physiologist, Alexander Gurwitsch, work- ing in his laboratory at Moscow, discovered that living matter gives off some type of energy which stimulates other living matter, near-by, to more active growth. The discovery of a new form of vital energy aroused much interest, and much opposition. There are people who are afraid of the word "vital" when used in biology; there is for them something mysterious about it, and they tell us that in science there is no place for mystery, we must adhere to cold facts. Let us do this then, and consider the discovery of Gurwitsch in terms of cold scientific facts. The original experiment of Gurwitsch was a simple one: he placed the root of an onion in a normal vertical position, then he pointed the root of another onion, held horizontally, directly toward the tip of the first root and very near to it. After some hours the vertical root was killed, cut, and stained, and its cells examined. It was found that the vertical root showed an increase in the number of cell divisions on that side which had been ex- posed to the horizontal root; in other words, the one root had given off some form of stimulus which had caused the other root to divide more on the one side than on the other. Now, onions give off volatile oils, and, therefore, probably have a strong influence on tissues. Perhaps it was merely some such chemical substance which produced a stimulation of cell division- there are many chemical substances which will stimulate growth— but the two onion roots were not in contact, there was only air between them. In order to test this question further Gurwitsch placed glass and quartz plates between the two roots, and, in the case of quartz, the increased growth in the one root still took place, that is to say, the energy, whatever its nature, will pass through a quartz plate. Now, ultraviolet rays pass through quartz, but not through glass, and for this reason Gurwitsch thought these vital rays to be comparable to ultraviolet rays. Whether or not these two forms of radiant energy, the Gurwitsch vital rays, and the ultraviolet rays, are identical, it is impossible to say; they appear to do the same thing, but until we know the precise nature of the vital radiant energy, it will be well to honor the discoverer and use his name to characterize the vital rays. The universal occurrence of the Gurwitsch rays in living matter is shown by the variety of sources from which they come. So far, the follow- *I am indebted to Mr. Lawrence S. Moyer for much information from the literature. 34 ing cells, organisms, and tissues have been shown by experiment to be the source of this form of energy: bacteria, the eggs of lower animals, tadpoles, yeast, plant seedlings, potatoes, beets, the blood of the frog and the rat, muscle, and cancerous tissue, all radiate energy which will increase the rate of division of other cells. The situation in the case of the tadpole is rather amusing. A mash made from the heads of tadpoles emanates rays which accelerate growth, but a mash made from any other part of the tadpole's body will not do this. If the entire living tadpole is used, and if it is pointed head first toward a plant root, then increased growth in the root will take place, but if the tadpole is turned around and pointed tail first, nothing happens. We can regard the existence of Gurwitsch rays as experimentally proven. Let us now turn to a purely theoretical consideration of facts which lead us to believe that vital rays of such a sort must exist. Man has long been aware of his radiation environment; he has known that he is sensitive to the rays of the sun, that they not only give him light and heat, but tan his skin. When sunlight is broken up into its com- ponent parts, we obtain a rainbow or spectrum. Beyond the visible blue end of the spectrum are the ultraviolet rays. These are very short, very active, very penetrating, and, in moderation, very beneficial. Man, and all living matter, appears to be very sensitive to the ultra- violet rays. If you have blistered your lips at the Grand Canyon on a very clear day, the ultraviolet rays from the sun are responsible. If your physi- cian has recommended pure sunlight for your room, it is the ultraviolet light to which he wants you exposed. When a child is put out into the sun as a cure for rickets, it is the ultraviolet rays which enter the body and effect the cure. While ultraviolet rays seem to be especially effective in maintaining health m man, there are other forms of radiation to which we are also sen- sitive, and which may exercise far more influence upon us than we realize. The most recently discovered rays are those which come from the depths of the open spaces between the stars. These are the so-called cosmic rays and represent excess energy given off in the formation of matter far out in interstellar space. The rays are very powerful; they penetrate 200 feet of water. While no definite biologic effect of cosmic rays has yet been found, it has already been suggested that man's susceptibility to cancer is greater in times of less cosmic radiation. Whether this is true or not cannot be said, but man is most certainly sensitively and delicately adjusted to so powerful a form of radiant energy. Familiar to every one are X-rays, another form of emanation which comes from a vacuum tube within which an electric discharge is taking place. These rays also affect living matter. 36 SCIENCE EDUCATION [Vol. 16 No. 1 Equally familiar are the emanations from radium. This heavy metal gives off three kinds of radiation: alpha particles, beta particles, and gamma rays. Radium emanations will quickly kill a living cell, or slowly poison a worker, and, in rare instances, effect cure, as in the case of can- cer, which means that living matter is very sensitive to radium emana- tions. While this type of emanation does not appear to play an important part in our everyday radiation environment, its influence may yet be great. Out in California, scientists have shown that fruit-flies allowed to breed in a locality where there is more earth radiation than elsewhere, develop, twice as often as do other flies, a tendency which causes the males to die before hatching. Radioactive mineral deposits which lie near the surface of the earth are the source of this radiation, to which, presumably, all liv- ing things are susceptible. If we recognize, as we must, that living matter is sensitive to its radia- tion environment, then we must also grant that these forms of radiant energy have materially influenced the course of evolution. If the sun, interstellar space, electric discharge, and radium are sources of radiant energy, is it unreasonable to assume that living matter, more complex than any of these, is also a source of radiant energy; is it un- reasonable to assume that living matter which takes in, and is itself so sensitive to radiant energy from the outside, should not, in turn, give off some of this energy? That a living animal gives off heat has never seemed a remarkable thing, for it is of such common experience. That animals give off light and electricity, is more extraordinary, yet we unhesitatingly admit it be- cause we must, for we know of light from the fire-fly and the glow-worm, and of electric shocks from the ray-fish. It is but a step from heat, light^ and electricity, to those other forms of radiant energy, ultraviolet rays, X-rays, and radium emanations. Let us approach our hypothesis, that living matter gives off radiant energy, from another angle. One of the chemical constituents of protoplasm is potassium. Another metal which has been found in plants is rubidium. Both potassium and rubidium are slightly radioactive. Knowledge of the presence of these elements in living matter is of long standing. But we have newer and better evidence. Recent work at the Russian Radium In- stitution has shown that radium, in almost infinitesimal amounts, has been found in living plants and animals. Is protoplasm less radioactive than the radioactive elements which it con tarns? We have, in this purely a priori evidence, reached the point where we can no longer doubt that living matter gives off radiant energy. And this is the opinion of an eminent group of biologists, chemists, and physicists who very recently met at the Kaiser Wilhelm Institute in Berlin and, in Oct. 1931] RADIANT ENERGY FROM LIVING MATTER 37 the face of much opposition, concluded that onion roots emanate a form of radiant energy, the exact nature of which no one cared to suggest. Then why is it, that many scientists still do doubt the existence of the Gurwitsch rays? Opposition seems to rest, not so much on objection to the experimental work, as on the very natural tendency of the scientific mind to react unfavorably toward anything mysterious. This is a weakness which is especially evident in those who view life in a purely mechanistic way. But, we say to them, the Gurwitsch rays are as physical, as ma- terialistic, as any phenomenon could be. The presence of radiant energy in protoplasm is one of the least mysterious things about living matter. What of those other properties of protoplasm, concerning which we know so little, its movement, its growth, its reproduction? Nothing in the living or non-living world is more mysterious than these happenings; yet this does not, necessarily, make them less physical, nor less chemical, nor mean that we shall not, some day, understand them. The only intelligent way to view a discovery which is shrouded in a veil of mystery is not to fear it, nor shun it, but to investigate it by scientific methods, and this is what Gur- witsch did when he attempted to prove that vital rays are of the same general nature as ultraviolet rays. If we simply do not like an idea, that is no reason for casting doubt on its truth. There are men still who do not like "evolution." Such feelings are very human; man has always disliked anything new if it upsets old and cherished beliefs; as Goethe put it: Art thou so weak, disturbed by each new word? Wilt only hear what thou'st already heard? To wondrous things art thou so used already. Let naught, howe'er it sound, make thee unsteady ! As for the Gurwitsch rays, let us keep an open mind; we may be wrong, but all evidence, experimental and theoretical, is with us, and of this we can at least be certain, that protoplasm is alive^ it is a dynamic and not a static system, and, as a source of energy, it has not an equal. [Reprinted from the Journal of Ecology, Vol. XIX. No. 2. August, 1931.] [All rights reserved.] SKETCHES OF THE VEGETATION OF SOME SOUTHERN PROVINCES OF SOVIET RUSSIA By WILLIAM SEIFRIZ. Introduction. Knowledge of plant life in the southern countries of the Union of Soviet Socialistic Republics^ is accessible to the outside world in the form of several classical Russian and German works, but these are none too familiar, and they leave some interesting territory untouched. Furthermore, Russian taxonomists and ecologists have been very active since the Revolution and have brought together many new facts. It seems, therefore, worth while to present, in English, an account of the flora of certain southern Soviet provinces, and thereby bring together some of the newer researches now scattered in relatively inaccessible journals, to which are added a few original observations of my own. I had the pleasure, in the summer and fall of 1929, of leisurely journeying from the Crimea to within 100 miles of the Chinese border in Turkestan. The present paper is the first of a series in which the botanical results of this excursion are described. The regions visited were the Crimean Mountains, the Georgian Military Way (North Caucasus), the Bakuriani Basin (Minor Cau- casus), the desert at Repetek (Kara Kum), and the Transilian Mountain range of Kazakstan (eastern Turkestan). My sojourn in these southern states of the Soviet Union was made in company with a number of Russian botanists who either travelled with me for a time or resided in the places where I stopped. I am indebted to them all and I shall express my appreciation in these articles; but to certain of these companions special thanks are due. I am grateful to Prof. N. A. Maximov who gave to my stay in Russia the necessary official backing; to Mrs Tatiana Krasnosselsky Maximova, who, with characteristic initiative and energy, helped our expedition through many trying experiences, and who has since efiectively obtained for me much information necessary to the satis- factory completion of these articles; and to Prof. Venedict Kolesnikov who was my companion in the Crimea and North Caucasus. I am also grateful to Dr Forrest Shreve, of Tucson, Arizona, for his courtesy in reading the manuscripts of these articles, and to Mr Paul Bausch for kindly making a number of drawings, including Fig. 3 of the present article. 1 Many encyclopaedic and geographic references give "Union of Socialistic Soviet Republics^ as the name of the present Russian Union. The Russian bank notes have Union of Soviet Socialistic Republics'* printed upon them. 361 William Seipriz I. THE ALTITUDINAL DISTRIBUTION OF PLANTS ON THE CRIMEAN MOUNTAINS (With Plates XIV -XV II and three Figures in the Text.) The success of the botanical study of the Crimea here presented is largely due to Mr V. F. Vasiliev, upon whom rested the final identification of all plants collected. Mr Vasiliev, with typical Russian courtesy, planned and conducted the trip into the mountains. His published accounts^ of the Crimean flora have been of great help in the writing of this article. I am further indebted to Mr N. V. Kovaliev, Director of the Nikita Gardens, for officially assuming half of the responsibility of the trip. The flora. Plant geography is an old subject. The evident and superficial things have been done. To add anything of fundamental significance now requires years of intensive and extensive work which must be left to those residing in the country. The chance visitor can grasp only something of the outstanding botanical features of the region; his glimpses assume a more substantial nature, and become of some real value, only when he has the help of the taxonomic specialists of the country. The Crimea, or Krim, is a peninsula projecting into the Black Sea from the south coast of Kussia. A very narrow strip of land saves the Crimea from being an island which it more resembles. It is of irregular shape, considerably broader than long. The 45th parallel passes across the peninsula, dividing the Crimea into two geographically distinct regions, the southern mountainous coastal strip, 30 miles in width, and the broad northern steppe which con- stitutes more than two-thirds of the total area of the state. The capital, Simferopol, is a few miles south of latitude 45°. The Crimean Mountains attain their maximum height close to the coast (Fig. 1). Here they rise to an altitude of 1500 m. within 8 km. of the shore. At one point an altitude of 1200 m. is reached within 3 km. of the shore; this is the peak Ai-Petri. It is impossible to refer to individual mountains in the coastal range, as the top is one extensive plateau known as the Yaila. The main portion of the plateau is some 30 km. in length, varying from J to 5 km. in width. The highest point, with an altitude of 1540 m., is toward the eastern end, just above the town of Yalta 2. * "Die Vegetationsverhaltnisse der Gegend Sudak-Aluschta" (published in Russian with a German summary). Journ. Gov. Bot. Oard. Nikita, Yalta, Crimea, 10 (2), 1928. 2 The spelling of Russian words has been much influenced by German translations and by historical traditions. The transliterations adopted here aim not only at correct pronunciation, but also at retaining the Russian spelling in so far as this is possible. Pronunciation is of greater importance than spelling, and where it is necessary to change the latter to achieve the former, I have done so. There are naturally certain minute shades in pronunciation which only Russians Vegetation oj some Southern Provinces of Soviet Russia 302 Crimea's coast has long been famed as Russia's riviera (Fig. \). The largest of the south-coast towns is Yalta. Several miles east of Yalta are the Nikita Botanic Gardens, consisting of 500 acres extending up from the shore to the village of Nikita. Immediately above Nikita rises a high spur which projects southward toward the coast from the main Yaila plateau (Fig. 1). The journey into the mountains leads from the Nikita Gardens up along the west slope of what I shall call the Nikita spur, over the Yaila plateau, and part way down the north side of the coastal range to the former monastery of Kozmo Damian where the night is spent. The return trip may be made by way of the east slope of the spur. 5EVAST0P0 UU5HTA GURZUF IMTA QARDEN-S YAL.TA Fig. 1. The southern tip of the Crimea showing the Yaila plateau. The Crimea lies in the southern part of the temperate zone with only the suggestion of a tropical flora. Ripe figs, Ailanthus, and the leguminous tree Albizzia, together with a hot and dry summer, remind one of the Near East, but the absence of cultivated palms and the scarcity of citrus fruits indicate that the Crimea has a winter not tropical in nature. The average summer (August) temperature at Nikita is 24-6'^ C, and the winter (January) tern- can accomplish. It seems best, for English readers, to use " j " for the sound of "s" in "pleasure,'* and "y" for the Russian H, the sound in the pronoun "ye": thus "Yalta" and not "Jalta" for the Crimean seaside town. Whether to use "v," "ff" or "w" for the familiar ending to many Russian words is a question which is answered as one wishes. The Russian letter which ends words of this sound is "b." This is not our "b," for which the Russians have another letter, but our "v." Since "v" gives the sound fairly well, it is the best of the three possible transcriptions, as it conforms not only to pronunciation but to Russian spelling. For the same reason 1 much prefer "Buchara" for the Turkestan city to the familiar English transcription " Bokhara," which gives neither the Russian spelling nor the Russian pronunciation. The addition of "c" m "Buchara" is necessary to approach more closely to the Russian pronunciation. 363 William Seifriz I m tl perature 4° C, The rainfall at sea-level on the southern coast (Nikita) is 50 cm. a year (on the northern steppes it is but 30 cm.) and on the mountain tops, the high Yaila plateau, it is 120 cm. The yearly distribution of rainfall in the Crimea is irregular. On the steppes in the north, summer rains are con- tinental in character and constitute 43 per cent, of the total annual amount. In the mountains, the yearly distribution is more uniform, with spring and summer maxima. On the southern coast, 56 per cent, of the total annual precipitation falls from October to February and 24 per cent, from May to August. So dry a spring has a marked influence on the character of the vegetation of the coast. The Crimean mountains rise abruptly from the south coast, leaving but a narrow strip of land suitable for cultivation. ReUcs of a former forest are still present here, but most of the area is cultivated, being planted chiefly with grapes and tobacco. This coastal agricultural strip constitutes the first of the five zones into which I have divided the southern slope of the Crimean mountains. Zone I. The coast (0-300 m.). Juniperus excelsa is the only tree which forms pure stands along the southern shore of the Crimea. Here and there small woods of a pure Juniperus growth still persist, left-overs from an extensive forest which has long since been cut. The juniper grows on poor rocky soil to the almost complete ex- clusion of other coastal trees such as the oaks. Five species of Juniperus occur in the Crimea, J. excelsa, J. oxycedrus, J. depressa, J . foetidissima and J. sahina\ the last is found on the higher mountain peaks. Of these species J. excelsa is by far the most abundant. J. oxycedrus is the only other juniper occurring in the first zone in any quantity and it is not frequently seen. This last species is of interest because it ordinarily exists as a low shrub, yet it may develop, when given time, into a substantial tree. (WulS^ gives a sixth species, J. isophyllos, as occurring along the coast with J. excelsa.) Two other trees, Quercus piibescens and Carpinus orientalis, join Juniperus excelsa in characterising the coastal region. Of these three only Juniperus is limited to the first zone. Quercus and Carpinus also occur further up if they can find space in areas deforested of pine and beech. Other oaks occurring in the Crimea are Q. sessiliflora at higher altitudes on the southern slope, and Q, pedunculata chiefly on the northern side of the mountains. These two species are very much alike. A second Carpinus, C. betulus, reaches the tree line. Acer campestre is a not infrequent inhabitant of the coastal region. A, hyrcanum is a high-altitude species. Fraxinus excelsior and occasionally F. oxycarpa are found in the first zone. Arbutus andrachne is a conspicuous tree of the coast because of its bark, which is smooth, of a rich reddish brown colour, and constantly peeling. The tree is at home in the Crimea and Asia Minor. 1 E. Wiilff . Vegetatiombilder aus der Krim, Jena, 1926. Vegetation of some Southern Provinces of Soviet Russia 304 There are a large number of wild pear, apple and cherry trees in the Crimea; Pirus elaeagnifolia is one of the most common of these. The closely related species P. communis may attain great age. Equally abundant is the wild apple, Pirus malus (Malus communis) ; somewhat less frequent is Prunus spinosa, and the wild sweet cherry Prunus avium. Chlorosis is of unusual occurrence among the fruit trees of the Crimea, both wild and cultivated forms, especially the pears, due to a strong alkaline soil. Pistacia mutica (Anacardiaceae), the turpentine tree, though but occa- sionally seen, is a true representative of the flora of the Crimea, as it is also of that of Italy. The most historic representative of this species, at Nikita, was seen by Engler in 1900; he estimated its age as 1000 years. An interesting shrub is Paliurus spina Christi, one of the Rhamnaceae, a Mediterranean and Near East genus. The plant attracts attention because of the broad fringe or wing on the fruits. Occurring very abundantly along the coast is Ailanthus glandulosa. It is usually found as a rank-growing roadside weed some 4 to 6 ft. in height, but if given the opportunity it reaches tree size and forms a striking feature of the landscape when bedecked with its large clusters of red fruit in late August. The ubiquitous Rosa canina is abundant along the coast, extending up into the mountains. Among the herbaceous plants certain species grow so profusely that they conspicuously characterise the coastal vegetation. This is especially true of Clematis vitalba, the only species of this genus in the Crimea. It grows in great luxuriance. The plant is distributed not only throughout southern Russia and the Mediterranean region, but extends, as is well known, north-westwards through Europe to southern England. I was interested in finding Clematis abundant and in rich full flower keeping company with Artemisia and Alhagi in the Kizil Kum desert near the Aral Sea. Dr Shreve informs me that C. ligusticifolia is common in the Arizona deserts, with Atriplex occurring on the alkaline flood plains. Euphorbia biglandulosa is widely spread in the Crimea along the coast; in Greece it covers large waste areas. Along the shore grows Asphodeline lulea; A. taurica is another Crimean species of this genus. Iberis taurica (Cruciferae) is an endemic, occurring in stony areas. Numerous world-known herbaceous plants occur by Crimean roadsides. Cichorium intybus is a frequent reminder of American fields. Verbascum is represented by numerous species. Leguminous plants, such as Spartium junceum, are abundant, as also the Cruciferae. Five Papavers occur in the Crimea. They are regarded as weeds and are of wide distribution, occurrmg from the coast to alpine pastures. Ranunculus is well represented. The orange-coloured Crocus susianus is found along the coast near Nikita, with its cousin, the brilliant purple-flowered C. speciosus. Scilla autumnalis 365 William Seifrtz and Stachys iberica are other low-altitude forms. Ephedra vulgaris is at home here; it is found just south of the 45° parallel from Spain to eastern Turkestan. There are some fifty species of grasses in the Crimea, Festuca sulcata and Hordeum bulbosum being common at Nikita. Fifteen Carexes occur. That world-wide species, Equisetum arvense, is also present. Ferns are not numerous, only three genera and four species existing on the peninsula. Introduced plants which have become wild, together with those maintained by cultivation, often distinguish the floral topography of a country more than do the native plants. This is especially true in a densely populated region where intense agriculture is pursued. Exotic plants are the result of man's work, and man is usually omitted as a factor in ''natural" plant distribution. The landscape of the Crimean coast is more strikingly characterised by intro- duced and cultivated plants than by the native species. To omit mention of the tall and stately Cupressiis, of Cedrus, and Albizzia, would be to fail to give an accurate picture of the Crimean coastal flora. The symmetrical cypresses are conspicuous along the southern coast. Among the occasional exotics are the two superb cedars, Cedrus atlantica and C. libani. The legu- minous tree Albizzia {Acacia) julibrissin is extensively planted as a shade and decorative tree. It does well along city streets because of its drought- resistant qualities. When in flower it is most decorative. The tree is a native of Trans-Caucasia and China. Quercus suber is now being introduced with the ultimate intention of making Russia independent of Spain in its need of cork. Ripe figs, Ficus carica, add a subtropical touch. Punica granatum is likewise grown. The cultivated Citrus trifoliata is the only citrus plant in the Crimea. The two most extensive crop plants are tobacco and grapes. ''Dubek" is the Tartarian name for the finest of Russian tobaccos. It is grown in the Crimea. " Vinograd" or "wine bunches" as the Russians call grapes, constitute the chief agricultural product of southern Crimea. Vineyards cover many of the lower mountain slopes (PL XVI, Phot. 5). The scientific study and control of Crimean grape culture and wine production is in the hands of the Experi- mental Station at Nikita Gardens. A total of 800 varieties of grapes from all parts of the world are grown here. Of this number, 670 belong to the species Vitis vinifera. The grapes are converted into wine in the Station's own cellars, which have produced some of Russia's best wines for more than a century. Through the courtesy of the Director, Mr M. A. Gerasimov, the vineyard and cellars may be visited. The varieties of wines produced here are numerous, from the usual "vin ordinaire," to Malbeck, Bordeaux, Madeira, and the selected Muscat rouge de Frontingant, acclaimed the finest wine of the world, though another connoisseur gives preference to the equally soft and sweet Muscat noir d'Alican (Caillaba). Very fine also is Bastardo 26 from Spanish grapes ; its flavour is more distinctive and less sweet, though still soft. Russia I I T I ■I I f e f DURNAL OF ECOLOGY Vol. XIX, Plate XIV m to U > o «0 O PL, u > CO a C/5 ^ (D 2 o EIFRIZ — Sketches of the Vegetation of some Southern Provinces of Soviet Russia : I, Altitudinal Distribution in the Crimean Mountains Vegetation of some Soiithern Provinces of Soviet Russia 366 hopes to replace France in the production of Muscat owing chiefly to the destructive work of Phylloxera vastatrix in the latter country. One article of food commonly sold along the southern coast, but grown in north Crimea, is of interest to Americans as a contribution of their continent to Europe; it is *'corn on the cob." Children peddle it cooked, ready for eating. Still another plant which is widely grown is the sunflower; the seeds are eaten, though chiefly used for extracting an oil which constitutes the main cooking and table oil of Russia. Zone II. The Pinus laricio belt (300-800 m.). The Crimean pine, Pinus laricio var. pallasiana, forms a belt on the southern slope of the Crimean mountains at an average altitude of about 500 m. The association is a nearly pure one (PL XIV, Phot. 1). This pine rarely gets as low as the seashore nor as high as the plateau. The forest floor of the Pinus laricio association harbours little vegetation except for small patches of Dryopteris filix-mas, one of the few Crimean ferns. In open areas an occasional Pirus elaeagnifolia and P. communis are found. It is here that the oldest of the wild pear trees occurs, estimated to be over 200 years. Ruscus aculeatus grows in the more open areas of the woods. On the edges of the forest above the village of Gursuf one may find Saxifraga irrigua. Other herbaceous species occur in this zone, but they are more typical of the adjoining higher region. The second zone is essentially a pure stand of the indigenous Crimean pine which has received the name P. taurica, though it is perhaps better called by its older name. (Tauria is an ancient name for Crimea.) Many of the pine trees attain very good size and constitute the monarchs of the native flora on the south coast (PI. XIV, Phot. 2). Zone III. The upper herbaceous region (800-1000 m.). The third zone possesses the two characteristics necessary to form a good herbaceous undergrowth, open woods and ample moisture. This latter con- dition is maintained by abundant and hard rains, close-hanging clouds, and an absence of desiccating winds. (We recall that the annual rainfall at Nikita Gardens, at sea-level, is 50 cm., while on the Yaila plateau, at 1500 m., it is 120 cm.) Several new trees appear above 800 m. ; most of the lower altitude ones remain. Two of these new members characterise the arboreal vegetation of the third zone ; they are Pinus silvestris and Fagus taurica. The former, the most widespread European pine, is here intermixed with P. laricio var. pallasiana (P. taurica) from the second zone. In addition to these two species of pine, a third occurs elsewhere on the south coast of the Crimea, Pinus pithyusa var. stanhevici, a close relative of the Caucasian P. pithyusa. Face ^.366 ' DURNAL OF ECOLOGY Vol. XIX, Plate XIV f^ ^^ u > o is «/5 O 03 u 03 CO ^ o C/3 O u O ^ O EIFRIZ — Skp:tches of the Vegetation of some Southern Provinces of Soviet Russia : I, Altitudinal Distribution in the Crimean Mountains Vegetation of some Sofithern Prorinres of Soviet Russia Md hopes to replace France in the production of Muscat owing chiefly to the destructive work of Phylloxera vastatrix in the latter country. One article of food commonly sold along the southern coast, but grown in north Crimea, is of interest to Americans as a contribution of tlieir continent to Europe; it is ''corn on the cob." Children peddle it cooked, ready for eating. Still another plant which is widely grown is the sunflower; the seeds are eaten, though chiefly used for extracting an oil which constitutt^s the main cooking and table oil of Kussia. Zone II. The Pinus lartcio belt (300-800 m.). The Crimean pine, Pinus laricio var. pallasiana, forms a belt on the southern slope of the Crimean mountains at an average altitude of about 500 m. The association is a nearly pure one (PL XIV, Phot. 1). This pine rarely gets as low as the seashore nor as high as the plateau. The forest floor of the Pinus laricio association harbours little vegetation except for small patches of Drf/operis jilix-mm, one of the few Crimean ferns. In open areas an occasional Pirus elaeagnifolia and P. communis are found. It is here that the oldest of the wild pear trees occurs, estimated to be over 200 years. Ruscus aculeatus grows in the more open areas of the woods. On the edges of the forest above the village of Gursuf one may find Saxifraga irrigua. Other herbaceous species occur in this zone, but they are more typical of the adjoining higher region. The second zone is essentially a pure stand of the indigenous Crimean pine which has received the name P. taurica, though it is perhaps better called by its older name. (Tauria is an ancient name for Crimea.) Many of the pine trees attain very good size and constitute the monarchs of the native flora on the south coast (PI. XIV, Phot. 2). Zone III. The upper herbaceous region (800-1000 m.). The third zone possesses the two characteristics necessary to form a good herbaceous undergrowth, open w^oods and ample moisture. This latter con- dition is maintained by abundant and hard rains, close-hanging clouds, and an absence of desiccating winds. (We recall that the annual rainfall at Nikita Gardens, at sea-level, is 50 cm., while on the Yaila plateau, at 1500 m., it is 120 cm.) Several new trees appear above 800 m. ; most of the lower altitude ones remain. Two of these new members characterise the arboreal vegetation of the third zone; they are Pinus silvestris and Fagus taurica. The former, the most widespread European pine, is here intermixed with P. lancio var. pallasiana (P. taurica) from the second zone. In addition to these two species of pine, a third occurs elsewhere on the south coast of the Crimea, Pinus pithyusa var. stankevici, a close relative of the Caucasian P. pithyusa. Face ^.366 INTENTIONAL SECOND EXPOSURE 367 William Seifriz Fagus, more abundant on the north side, forms small pure stands at high altitudes on the south slope, especially on south-eastern exposures where there is good soil. The two pines are restricted to south-western rocky areas. The beech is so much more typical of the northern slopes of the mountain range, where it forms pure forests miles in extent, that we shall leave further consideration of it until later. A very few scattered specimens of the yew Taxus baccata are to be found in these high rocky regions. Another species of Carpinus appears in the third zone, C, hetulus, a tree more typical of central and western Europe where it is quite common. The widely distributed European oak, Q. jpedunculata, is also here at 900 m., but it is not restricted to this altitude. Another new species in this region is the maple A, hyrcanum. The northern and high altitude tree, Populus tremula, is scattered in the open areas of this zone, though it is not a typical Crimean tree. Fraxinus excelsior is still present. Cornus mas with its brilliant red berries may likewise be found. Its distribution is also very wide. To one familiar with south Russian plants, it would be a notable omission if Sorbus aucuparia were not mentioned as a high-altitude species. This small tree, which reaches the tree line throughout the Caucasus and again appears in the mountains of eastern Turkestan, is to be found in the Crimea but it is not abundant. It is a relic of the glacial age. The plant occurs in the third zone between 850 and 900 m. altitude. A second species of this genus, which is much more numerous in the Crimea, is Sorbus torminalis, so strikingly distinguished by its leaves from its cousin, S. aucuparia. This plant is rather common, and while growing best at 700-1100 m. altitude, it is to be found from the coast to, or near, the tree line. There are nine species of Sorbus in the Crimea, including Sorbus aria. Pinus silvestris and Carpinus betulus are the distinguishing trees of the third zone, but the arboreal vegetation characterises the zone less than does the succulent undergrowth. An open herbaceous zone at high altitude, sepa- rating a lower forest of large trees from a higher one of small trees, is generally characteristic of plant zonation on mountain sides; thus Shreve^ has found such a zone on the mountains of Jamaica and P in Java; we shall find the same at high altitudes in the Minor Caucasus. Among the herbs of Zone III the following are the most conspicuous in August : Atropa belladonna, with its glossy, black berries ; the European golden rod, Solidago virga-aurea, is another very striking and beautiful plant; Chrysanthemum parthenium; Campanula sibirica (and another larger species); the umbelliferous Laserpitium hispidum (its better known cousin, Daucus carota, is found at lower altitudes); two Centaureas, C.jacea and C. montana; ^ Forrest Shreve. "A montane rain-forest." Carnegie Inst. PubL 199 {1914:). 2 WHliazn Seifriz. "The altitudinal distribution of plants on Mt Gedeh, Java." Bull. Torrey Bot. Club, 50, 283 (1923). JOURNAL OF ECOLOGY Vol. XIX, Plate XV ' "K. ^nv*,, ^■t.>^> ! -V ^•^l A. • ::$m..^^: u.;/ ■«#^kt>i% Phot. 3. Pinus silvestris forming the tree-hne at 1200 metres. Si Phot. 4. Fagus taiivica creeping down on the south slope; summit of Yaila to the right. SEIFRIZ — Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: I, Altitudinal Distribution in the Crimean Mountains Vegetation of some Southern Provinces of Soviet Russia 368 the world-wide Epilobium angustifoUum (not a typical Crimean plant) ; Echium vulgare ; Cirsium incanum ; Linaria vulgaris (it is always of interest to find in far corners of the world species identical with those so familiar at home); Melampyrum nemorosum (Scrophulariaceae) ; Ranunculus polyanthemos; Salvia glutinosa, with its ingenious automatic spring for cross pollination ; the endemic, Melilotus tauricus] and Lappa {Arctium) tomentosa (Compositae), with huge (maximum 18 inch) heart-shaped leaves. One fern is met with, Dryopteris filix-mas. Zone IV. The tree line (1000-1250 m.). As one climbs on to the first shelf of the high Yaila plateau at about 1000 m. or more, there is a sudden change in the type of vegetation. The change may be clear cut or not, depending upon the topography of the imme- diate surroundings. Alpine plants make their first appearance. Trees are much reduced in size, more irregular in shape, and occur in smaller groups of close formation. Typical Krummholz is met with. Here occurs the tree line, formed by Pinus silvestris (PL XV, Phot. 3) and Fagus taurica (Phot. 4); the two species rarely occur together. On the western slope of the Nikita spur the pine alone forms the tree line. On the eastern slope the beech predominates. The fourth zone does not form a continuous belt, nor is it extensive in width. It consists of scattered areas where the trees of the third zone, more dwarfed in size, get up into the alpine pastures. The tree line region is essentially composite in character, consisting of small woods and meadows; floristically it is subalpine. Among the meadow plants the typically Crimean umbelliferous Seseli gummiferum and its cousin /S. lehmanni are to be found, as also the endemic Cachrys alpina and Cistus tauricus. Zone V. Alpine pastures (1250-1540 m.). The Crimean coastal mountains do not rise to peaks but culminate in flat meadows which form one long continuous plateau, the Yaila. Only an occa- sional small rocky summit is to be found. The Yaila is some 30 km. long with a maximum width of 5 km. and an altitude ranging from 1250 to over 1500 m. The few rocky mounds on top rise to maximum heights of 1526 and 1540 m. The open wind-swept meadows of Yaila form the alpine pastures of the fifth zone. The ground is well covered with grass and herbaceous species. Rock plants such as Saxifraga are not numerous. Before the Soviet Government put an end to pasturing on Yaila, these meadows were overrun by hordes of cattle, with the result that relics of the original Crimean alpine flora were to be found only on rocky summits. The plants are now gradually spreading to the floor of the plateau. Among them is the most striking and picturesque of the alpine flowers on Yaila, the Crimean "Edelweiss" Cerastium biebersteini, one of the Caryophyllaceae (PL XVI, JOURNAL OF ECOLOGY Vol. XIX, Plate XV Phot. 3. Pinits silucstris forming the tree-Hne at 1200 metres. Phot. 4. ragiis tauvica creeping down on tlic south slope; siunmit of Yaila to the right. SEIFRIZ— Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: I, Altitudinal Distribution in the Crimean Mountains 77/ Vegetation of some Southern Provinces of Soviet Russia 368 the world-wide Epilobium angustifolium (not a typical Crimean plant) ; Echium vulgaTe\ Cirsium incanum\ Linaria vulgaris (it is always of interest to find in far corners of the world species identical with those so familiar at home); Melam/pyrufri nemorosum (Scrophulariaceae) ; Ranunculus polyanthemos ] Salvia glutinosa, with its ingenious automatic spring for cross pollination ; the endemic, Melilotus tauricus) and Lappa (Arctium) tomentosa (Corapositae), with huge (maximum 18 inch) heart-shaped leaves. One fern is met with, Dryoptcris filix-mas. Zone IV. The tree line (1000-1250 m.). As one climbs on to the first shelf of the high Yaila plateau at about 1000 m. or more, there is a sudden change in the type of vegetation. The change may be clear cut or not, depending upon the topography of the imme- diate surroundings. Alpine plants make their first appearance. Trees are much reduced in size, more irregular in shape, and occur in smaller groups of close formation. Typical Krummholz is met with. Here occurs the tree line, formed by Pinus silvestris (PI. XV, Phot. 3) and Fagus taurica (Phot. 4); the two species rarely occur together. On the western slope of the Nikita spur the pine alone forms the tree line. On the eastern slope the beech predominates. The fourth zone does not form a continuous belt, nor is it extensive in width. It consists of scattered areas whc^re the trees of the third zone, more dwarfed in size, get up into the alpine pastures. The tree line region is essentially composite in character, consisting of small woods and meadows; floristically it is subalpine. Among the meadow plants the typically Crimean umbelliferous Seseli gummiferum and its cousin S. Ichmanni are to be found, as also the endemic Cachrys alpina and Cistus tauricus. Zone V. Alpine pastures (1250 1540 m.). The (Crimean coastal mountains do not rise to peaks but culminate in flat meadows which form one long continuous plateau, the Yaila. Only an occa- sional small rocky summit is to be found. The Yaila is some 30 km. long with a maximum width of 5 km. and an altitude ranging from 1250 to over 1500 m. The few rocky mounds on top rise to maximum heights of 152G and 1540 m. The open wind-swept meadows of Yaila form the alpine pastures of the fifth zone. The ground is well covered with grass and herbaceous species. Rock plants such as Saxifraga are not numerous. Before the Soviet Government put an end to pasturing on Yaila, these meadows were overrun by hordes of cattle, with the result that relics of the original Crimean alpine flora were to be found only on rocky summits. The plants are now gradually spreading to the floor of the plateau. Among them is the most striking and picturesque of the alpine flowers on Yaila, the Crimean "Edelweiss" Cerastium biebersteini, one of the Caryophyllaceae (PL XVI, INTENTIONAL SECOND EXPOSURE JOURNAL OF ECOLOGY Vol. XIX, Plate XVI [ W^ l' ; n u 369 William Seifriz Phot. 7). It first appears at the tree line (1250 m.) and extends to the highest rocky summits (1540 m.). : The Labiate Sideritis taurica, (PI. XVI, Phot. 6), an endemic, is also a frequent inhabitant of the plateau pastures. This Crimean native gets into the high Caucasus and Asia Minor. Viola altaica var. oreades, but sparingly found in flower in August, blossoms profusely over the plateau in May. It is a much sought after violet by western gardeners. Abundant also are the ' genera Alchemilla, Trifolium and Festuca. The northern slope. The north side of the Crimean coastal range has quite a different story to tell. Immediately on leaving the plateau meadows, one enters a dense beech forest, and this forest, an almost pure stand of Fagus taurica, extends for many miles, covering all the northern mountain slopes. It is a superb sight, whether one views it from above or from within. The tree line on the north side is slightly higher than on the south, creeping up to 1300 m. The beech trees at this altitude are low and bent (PI. XVII, Phot. 8), but they soon, at a lower altitude, become fine tall specimens closely grouped to form a dense forest (PI. XVII, Phot. 9). At 720 m. the trees attain their best growth with excellent boles 40-50 cm. in diameter and 30 m. or more in height. The beech forest descends to 500 m., to the foot hills north of which lie the vast steppes which make up most of the Crimea. The change in size of the beech, from its lower limit of 500 m. to the tree line at 1300 m., has been described by Poplawska. Fig. 2 is a modified copy of a drawing made by Poplawska^. The figure shows the change which the beech undergoes with change in altitude. The naming of the species of beech which covers the north side of the Crimean mountains has been an interesting problem now finally solved by Poplawska. The Crimean beech resembles F. silvatica of central Europe and was formerly regarded as of this species; it also bears a close resemblance to the Caucasian F. orientalis to which species it was later assigned. Poplawska finds that the Crimean beech has flowers as in F. orientalis, fruits as in F, silvatica, and leaves intermediate in size between these two species. Further- more, the Crimean beech develops root shoots as does its closer relative F, orientalis, which does not seem to be true here of F. silvatica. Poplawska, therefore, proposes the new specific name, Fagus taurica. Scattered along the fringe of the beech forest are several species from the southern side of the mountains. The two pines, P. laricio and, chiefly, P. siU vestris, occasionally occur, likewise examples of Carpinus betulus, Quercus pedunculata and Fraxinus excelsior, 1 Poplawska, H. "Die Buche in der Krim und ihre Variabilitat." Osterrekh. Bot. Zeitschr, T7, 23, 1928. See also ''ttude sur la variabilite du hetre de Crimee" (in Russian with French summary), Recherches sur les Pares Nationaux (Leningrad), 1926. Phot. 6. Sideritis taurica on the summit of Yaila. Phot. 5- The Crimean coast: pine on the left; junipers, oak, cedars and vineyards below. Phot. 7. Cerastium hiehersteini near the summit of Yaila. SEIFRIZ— Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: I, Altitudinal Distribution in the Crimean Mountains JOURNAL OF ECOLOGY Vol. XIX, Plate XVI 369 William Skikiuz Phot. 7). It first appears at the tree line (1250 m.) and extends to the highest rocky summits (1540 m.). The Labiate Sideritis taurica, (PL XVI, Phot. 6), an endemic, is also a frequent inhabitant of the plateau pastures. This Crimean native gets into the high Caucasus and Asia Minor. Viola altaica var. oreades, but sparingly found in flower in August, blossoms profusely over the plateau in May. It is a much sought after violet by western gardeners. Abundant also are the ' genera Alchemilla, Trifolium and Festuca. The northern slope. The north side of the Crimean coastal range has quite a different story to tell. Immediately on leaving the plateau meadows, one enters a dense beech forest, and this forest, an almost pure stand of Fagus taurica, extends for many miles, covering all the northern mountain slopes. It is a superb sight, whether one views it from above or from within. The tree line on the north side is slightly higher than on the south, creeping up to 1300 m. The beech trees at this altitude are low and bent (PI. XVII, Phot. 8), but they soon, at a lower altitude, become fine tall specimens closely grouped'to form a dense forest (PI. XVII, Phot. 9). At 720 m. the trees attain their best growth with excellent boles 40-50 cm. in diameter and 30 m. or more in height. The beech forest descends to 500 m., to the foot hills north of which lie the vast steppes which make up most of the Crimea. The change in size of the beech, from its lower limit of 500 m. to the tree line at 1300 m., has been described by Poplawska. Fig. 2 is a modified copy of a drawing made by Poplawska^. The figure shows the change which the beech undergoes with change in altitude. The naming of the species of beech which covers the north side of the Crimean mountains has been an interesting problem now finally solved by Poplawska. The Crimean beech resembles F. silvatica of central Europe and was formerly regarded as of this species ; it also bears a close resemblance to the Caucasian F. orientalis to which species it was later assigned. Poplawska finds that the Crimean beech has flowers as in F. orientalis, fruits as in F. silvatica, and leaves intermediate in size between these two species. Further- more, the Crimean beech develops root shoots as does its closer relative F. orientalis, which does not seem to be true here of F. silvatica. Poplawska, therefore, proposes the new specific name, Fagus taurica. Scattered along the fringe of the beech forest are several species from the southern side of the mountains. The two pines, P. laricio and, chiefly, P. sil- vestris, occasionally occur, likewise examples of Carpinus betulus, Quercus pedunculata and Fraxinus excelsior. 1 Poplawska, H. "Die Buche in der Krim und ihre Variabilitat." Osterreich. Bol. Zeitschr. 77, 23, 1928. See also "fitiide sur la variabilite du hetrc de Crimce" (in Russian with French summary), Becherches sur Us Pares Nationaux (Leningrad), 1920. hi Phot. 6. Sideritis taurica on the summit of Yaila. Phot. 5. The Crimean coast: pine on the left; junipers, oak, cedars and vineyards below. I'hot. 7. Ccrastiiim hieherstcini near the summit of Yaila. SEIFRIZ— Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: I, Altitudinal Distribution in the Crimean Mountains INTENTIONAL SECOND EXPOSURE OURNAL OF ECOLOGY Vol. XIX, Plate XVII o u O 03 u O -t-i o r-' o o •^1 u O a.) CD O O ^ SEIFRIZ— Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: I, Altitudinal Distribution in the Crimean Mountains Face > OURNAL OF ECOLOGY Vol. XIX, Plate XVII -' -.» ^i-f^^, ^\,'. '#.'« «••»»«»•. ^^ «jNPf ■"«r' ^"V «r,^'»-' •'•r^"f» '^ ^H. ' '.'f^^ • ''%* ■ .^•;.i ■»>'•. :'!«'»' 7) O CD O o o o N o I— H o I '-r/;if '•T \ -.v ' %^Sr ^ k\ W- 5 S: iA . , * ■^... O U b • Q I o ^ o s .1-1 -.s* « ° CO ;S O S I > to O 0. < --fS-.^' .^}*?*;'f SEIFRIZ Sketches of the Vegetation of some Southern Provinces OF Soviet Russia : II, Plant Life along the Georgian Military Way 378 Veyetation of some Southern Provinces of Soviet Russia Turkey. The montana form of the pine along the Georgian Military Way has been recognised by Fomin and Radde^ as P. silvestris var. alpina. The dis- tinction, though taxonomically doubtful, is stamped ecologically by soil preference; the montana form occurs on granite. The only other conifers along the Georgian Military Way are three junipers and a Taxus. Here, in the second forest zone (Zone IV), Juniperus oblonga occurs as a shrub, with Taxus baccata. We shall meet with another juniper higher up. Picea, so typical of the Minor (South) Caucasus, where it forms beautiful specimens in extensive forests, is restricted to the western parts of the High Caucasus, both in the north and the south, Tiflis being the eastern limit. There is no spruce on the Georgian Military Way. Flowers continue to flourish in small open areas. In the wet soil along the road the rich purple spike of Lythrum salicaria var. intermedia stands in great numbers. Radde cites L. hyssopifolia as occurring in the Caucasus, though I did not meet with it. Another blue spike is that of Salvia nemorosa. Campanula rapunculoides, Picris strigosa and Aster ibericus are pretty addi- tions to the roadside flowers. Zone V. The Daryal Canyon (1300 m.). The Daryal Canyon ofEerslittle encouragement to plants (PI. XVIII, Phot. 1). In its most precipitous parts its width is barely more than that of the road and the narrow river bed combined. Its walls rise almost perpendicularly to tremendous heights. The sun enters for but a few hours each day. Yet, in its more open parts, especially near the upper end, some plants manage to exist on the rocky ledges. The montana form of the pine is to be seen perched high on rocky crags (PI. XVIII, Phot. 3). Where a little soil has gathered, clumps of Juniperus sabina and J. oblonga find a foothold, so also Berberis georgica and Euphorbia, Sempervivum caucasicum flourishes well, tightly rooted in rock crevices, as is the habit of this genus. Here we also find a species of Scabiosa, the small S, bipinnata, and Pyrethrum, a less familiar genus among the asters, which occurs as the delightfully jaunty P. parthenifolium, clinging high where the precipitous cliffs of the Canyon open up into the sunshine (PI. XX, Phot. 6). We shall meet with the more abundant P. niveum later. A third species, P. macrophyllum, occurs in the Minor Caucasus. Here, at the south end of the Canyon, I collected the first fern, an Asplenium trichomanes. But three mosses were collected from the rocky clifis along the road coming out of the Canyon. These have been kindly identified for me by Mr K. S. Williams of the New York Botanical Gardens; they are Mielichhoferia nitida, Dicranum bonjeani and Sphagnum capillaceum. The first is an old world moss and the two latter are familiar North American species. The mammoth truncated cone of rock, which is crowned by the ruins of 1 Radde, G. Pflanzenverhreitung in den Kaulcasusldndern {Die Vegetation der Erde III). Leipzig, 1899. JOURNAL OF ECOLOGY Vol. XIX, Plate XVI II !^-!3<#*' %at!S' km.^ ^*^,^<---.- ••^..'^v.^ ■ -^^^y^'i^ •■ K. 03 H C O o U o Oh o r- -.1 ^ o I o :^ o ^ O o o s o 03 V2 A. '^1 SEIFRIZ Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: II, Plant Life along the Georgian Military Way INTENTIONAL SECOND EXPOSURE I m r,' n William Sbifriz 379 an ancient castle accredited to the legendary Queen Tamara, marks the upper (southern) limit of the Daryal Canyon and the beginning of the sixth (second herbaceous) zone. Zone VI. Teansition (1500 m.). The Castle of Tamara stands like a sentinel before the southern entrance of the Daryal Canyon (PI. XVIII, Phot. 1). Its position was undoubtedly selected because of strategic advantages. From this point on, the Georgian Military Way passes through high open treeless valleys, where the meadows though small are luxuriant with flowers. Herein lies the chief difference be- tween the first and this, the second, herbaceous zone; the former has much of the steppe in its make-up, the fields are extensive, and the vegetation is chiefly grass, but in the second herbaceous zone of the upper mountain regions the meadows are small and covered with subalpine flowers. Shortly beyond the upper end of the Daryal Canyon is the village, if such it may be called, of Gvileti. The altitude is 1500 m. To the west lies a valley which leads to the north-east glacier of Mount Kasbek. The surrounding country is very picturesque and constitutes one of the richest coUectmg grounds in this part of the Caucasus. At least two days should be spent at Gvileti ; one may spend the night at the primitive hostelry. Eight kilometres beyond Gvileti, at an altitude of 1705 m., lies the village of Kasbek. It is a mountain settlement of considerable size with a magnificent view of Mount Kasbek whose glistening dome rises to a height of 5043 m. (16,547 ft.). The older settlement at Kasbek is a primitive but unique little vill'age. The valley is small and offers little opportunity for agriculture, but the natives manage to raise some wheat and cattle. The valley leads to the eastern glacier of Mount Kasbek. The fields at Gvileti are uncultivated and therefore little disturbed by man. A mountain lake there adds an interesting plant to the flora, Phra^mUes communis, which grows profusely (PL XIX, Phot. 4). Small compact mounds of Juniperus oblonga are scattered about over the rocky floor (PI. XV 111, Phot. 2). Spiraea hypericifolia var. subalpina grows abundantly on the chffs, and with it Allium ruprecMii. , i i- -4. j The fields at Gvileti and Kasbek (1500-1700 m.) constitute the limited region of our sixth zone. At Gvileti, the ground is mostly rock, with sonae grass, scattered prostrate junipers, and an occasional Centaurea At kasbek subalpine flowers creep down into the grassy meadows between the cultivated fields. Here occur small woods of Populus tremuU (PI. XIX, Phot. 5). ihe poplars are relics of a former extensive forest and still exist because they have enjoyed the protection of the church. . The sixth zone is a transitional one, joining the lower rocky slopes to the higher subalpine fields. It is a zone characterised more by its geologic features than its sparse floristic ones. 1 ' h 1 I 380 Vegetation of some Southern Provinces of Soviet Russia Zone VII. Subalpine (1700 m.). Above Gvileti lies a subalpine region of great interest and beauty (1700- 2200 m.). Small trees, chief among which is the birch, alternate with flower- covered meadows bounded above by Rhododendron and alpine pastures. At Gvileti, Betula mddeana climbs to 2400 m. and gets down to 1700 m., occurring, therefore, in both the alpine and subalpine regions (PI. XX, Phot. 7). The more typical birch of the alpine zone is Betula pubescens, I am not quite certain in regard to a third kind of birch, B, pubescens var. raddeana, occurring at Gvileti. It appears to differ from both the pubescens and raddeana species and therefore was identified as a third form, though it may be identical with one or the other of the two species after which it is named. An interesting tree, of which I saw only one specimen in these higher mountain valleys, is the maple, Acer trautvetteri, which was growing at 2000 m. Radde says that this species may reach the tree line. Here at Gvileti it is near the tree line, but I never found it there in the Minor Caucasus, where it is very abundant, occurring just below the open subalpine woods of birch. Sorbus aucuparia begins to make its appearance at 1600 m., getting up into the alpine region at 2400 m. (PL XX, Phot. 7). This small tree is of very wide distribution in the mountains of the Soviet Union from the Crimea to Eastern Turkestan. It is a companion to the birch. Whether to regard the two as alpine or subalpine is purely a matter of defining the limits of these zones in that particular region where we happen to be at the time. In the Crimea, neither of these two high altitude trees plays a prominent part in the plant life of the mountains, they occur only sparingly, while in the Minor (South) Caucasus (Bakuriani) they grow in great abundance, form the tree line, and on certain slopes occur at high altitudes to the exclusion of all other arboreal forms. Usually, the tree line of birch and Sorbus makes a convenient upper limit of the subalpine zone, but where meadows of tall, showy, subalpine plants, such as Scabiosa, Del- phinium, Cephalaria and Epilobium, grow above the birches, then, naturally, we cannot regard the latter as delimiting the alpine and subalpine flora; furthermore, where the birches get so far above the fields of herbaceous succulents as to mingle with Rhododendron, as here at Gvileti, they become alpine, for their associates are not only Rhododendron but such other truly alpine forms as Antennaria and Sibbaldia. One other tree occurring here at 2000 m., though not abundantly, is Salix caucasica. The trail at Gvileti, with Mount Kasbek and its glacier constantly in view, rises but little above 2000 m. before it descends again to go into the rocky gorge at the head of which is the terminus of the glacier. The trail reaches its highest point in a field of flowers of great luxuriance and beauty. This sub- alpine meadow extends from 2000 to 2200 m., and is almost entirely covered with a tall rank growth of herbaceous plants : very little grass is visible, and lOURNAL OF ECOLOGY Vol. XIX, Plate XIX Phot. 4. Phragmites communis at Gvileti. Phot. 5. Populus tremula at Kasbek. 3EIFRIZ— Sketches of the Vegetation of some Southern Provinces Soviet Russia: II, Plant Life along the Georgian Military OF Way 380 Vegetation of some Southern Frovmces of Soviet Russia Zone VII. Subalpine (1700 m.). Above Gvileti lies a subalpine region of great interest and beauty (1700- 2200 m.). Small trees, chief among wliich is the birch, alternate with flower- covered meadows bounded above by Rhododendron and alpine pastures. At Gvileti, Betida raddeana climbs to 2400 m. and gets down to 1700 m., occurring, therefore, in both the alpine and subalpine regions (PL XX, Phot. 7). The more typical birch of the alpine zone is Betula pubescens. I am not quite certain in regard to a third kind of birch, B, pubescens var. raddeana, occurring at Gvileti. It appears to differ from both the pubescens and raddeana species and therefore was identified as a third form, though it may be identical with one or the other of the two species after which it is named. An interesting tree, of which I saw only one specimen in these higher mountain valleys, is the maple, Acer trautvetteri, which was growing at 2000 m. Eadde says that this species may reach the tree line. Here at Gvileti it is near the tree line, but I never found it there in the Minor Caucasus, where it is very abundant, occurring just below the open subalpine woods of birch. Sorbus aucuparia begins to make its appearance at 1600 m., getting up into the alpine region at 2400 m. (PL XX, Phot. 7). This small tree is of very wide distribution in the mountains of the Soviet Union from the Crimea to Eastern Turkestan. It is a companion to the birch. Whether to regard the two as alpine or subalpine is purely a matter of defining the limits of these zones in that particular region where we happen to be at the time. In the Crimea, neither of these two high altitude trees plays a prominent part in the plant life of the mountains, they occur only sparingly, while in the Minor (South) Caucasus (Bakuriani) they grow in great abundance, form the tree line, and on certain slopes occur at high altitudes to the exclusion of all other arboreal forms. Usually, the tree line of birch and Sorbus makes a convenient upper limit of the subalpine zone, but where meadows of tall, showy, subalpine plants, such as Scabiosa, Del- phinkim, Cephalaria and Epilobium, grow above the birches, then, naturally, we cannot regard the latter as delimiting the alpine and subalpine flora; furthermore, where the birches get so far above the fields of herbaceous succulents as to mingle with Rhododendron, as here at Gvileti, they become alpine, for their associates are not only Rhododendron but such other truly alpine forms as Antennaria and Sibbaldia. One other tree occurring here at 2000 m., though not abundantly, is Salix caucasica. The trail at Gvileti, with Mount Kasbek and its glacier constantly in view, rises but little above 2000 m. before it descends again to go into the rocky gorge at the head of which is the terminus of the glacier. The trail reaches its highest point in a field of flowers of great luxuriance and beauty. This sub- alpine meadow extends from 2000 to 2200 m., and is almost entirely covered with a tall rank growth of herbaceous plants: very little grass is visible, and INTENTIONAL SECOND EXPOSURE OURNAL OF ECOLOGY Vol. XIX, Plate XIX Pilot. 4. Phvagmites communis at Gvileti. Phot. 5. P'^piilifs tvciuida at Kasbek. '.EIFRIZ-Sketches of the Vegetation of some Southern Provinces OF Soviet Russia: II, Plant Life along the Georgian Military Way II William Seifriz 381 this emphasises Timofeev's contention for the Minor Caucasus, that alpine regions contain thirty to forty, and subalpine seventy, to eighty, species of herbs as compared with eight of grasses. The plants of the Gvileti and Kasbek valleys are so much alike that it will give a more unified picture to list them together. The following are the species collected in these two localities at the beginning of August. The final identification of them has rested with Mr Dmitri Sosnovsky, of the Tiflia Botanic Garden, to whom my thanks are due. PpLYPODlACEAE Asplenium trichomanes Athyrium alpestre Cryptogramme crispa Dryopteris oreades Dryopteris robertiana Polypodium vulgare var. rotundatum Woodsia f ragilis Orchidacbae Gymnadenia conopea Salicaceae Populus tremula Polygonaceae Oxyria digyna Rumex alpinus Chenopodiaceae Chenopodium foliosum Caryophyllaceae Alsine imbricata Dianthus alpinus Dianthus petraeus Minuartia imbricata Ranunculaceae Aconitum caucasicum Aconitum nasutum Aconitum orientale Delphinium flexuosum Ranunculus boissieri Papaveraceae Papaver bipinnatum Cruciferae Thlaspi arvense Crassulaceae Sedum oppositifolium Sempervivum caucasicum Saxifragaceae Pamassia palustris ROSACBAE Alchemilla pubescens Alchemilla tredecimloba Fragaria elatior Rosa dumetorum Rubus idaeus Rubus saxatilis Leguminosae Medicago glutinosa Vicia alpestris Geraniaceae Geranium pyrenaicum CiSTACEAE Helianthemum chamaecistus Thymelaeaceae Daphne glomerata Onagraceae Chamaenerium palustre Epilobium algidum Epilobium nervosum Umbelliferab Astrantia biebersteini Astrantia maxima Ericaceab Vaccinium vitis idaea Oleacbab Ligustrum alatum Gentianacbab Gentiana asclepiadea Gentiana caucasica Grentiana septemfida Labiatae Betonica grandiflora Mentha silvestris Thymus serpyllum SOLANACBAB Physalis alkekengi Scrophulariaceae Rhynchocorys orientalis Verbascum orientale DiPSACACEAB Cephalaria procera Scabiosa caucasica Campanulacbab Campanula collina Campanula rapunculoides Podanthum amplexicauli COMPOSITAB Centaurea fischeri var. ochroleuca Centaurea phrygia Doronicum macrophyllum Inula glandulosa Inula Selenium Leontodon hispidus Mulgedium racemosum Pyrethrum niveum Pyrethrum parthenifolium Senecio candolleanus Senecio nemorensis Solidago virga aurea Swertia iberica v. alleida Tanacetum vulgare Tragopogon reticulatus m ji ■ *> ^83 Vegetation of some Southern Provinces of Soviet Russia f"'" The most beautiful of the blossoms is Scabiosa caucasica, the queen of all the' Caucasian flowers. . . ;• ; 0* c\ ; ZoneVIII. Alpine (2200 m.). ' 'At 2200-2400 m., in the Gvileti valley, clinging to the precipitous north Slopes of the mountain, grows a dense thicket of Rhododendron caucasicum. Intermixed with it are Vaccinium vitis idaea, Sorbus aucuparia and Betula pubescens. The birch, in places, occupies the ground alone. An occasional inhabitant of these higher slopes is, again, Salix caucasica. The Georgian Military Way leads on from Kasbek to Kobi and to the Divide. The latter is a treeless pasture covered with grasses and the poisonous Veratrum album. The Caucasian lily, Lilium monadelphum, is also present. From its maximum altitude of 2345 m. (7692 ft.) the Way winds down through country less magnificent than the north slopes of the Caucasus, be- coming more and more arid until the semi-desert plains of Tiflis are reached at an altitude of 400 m. [Reprinted from THE AMERICAN JOURNAL OF PHARMACY, Philadelphia, Pa. April, 1932] THE INTRODUCTION OF LOBELIA SYPHILITICA INTO MEDICINE By Rodney H. True SOME years ago while looking up the story of the old preacher- physician, Jared Eliot, of Killingworth (now Clinton), Connecti- cut, I encountered his medically more eminent son-in-law, Dr. Benja- min Gale, perhaps his favorite pupil. Dr. Gale had a widespread reputation in the northern colonies, as is made clear by a correspond- ance that I had the good fortune to unearth some years ago in the Library of Congress at Washington. It is generally known that the use of Lobelia syphilitica L. against venereal disease is traced back to the New York Indians as appears in a communication from the Swedish botanist and traveler, Peter Kalm. Since certain details can be traced in the correspondence above alluded to, it seems to be worth while perhaps to bring it to light. It opens with a letter from Dr. Benjamin Gale, dated October 26, 1767, at Killingworth, addressed to Rev. Samuel Johnson, of Stratford, Connecticut, who was supposed to have access to Sir William Johnson, member of the Governor's Council of the New York Colony, who was supposed to have full information on the subject of interest. Dr. Gale writes: "Reverend and Dear Sir: Reading the other day Dr. Haller's Medical Cases and Experiments, communicated to the Royal Academy of Sciences at Stockholm, among other things I find a remedy for the Venereal Disease discovered by Peter Kalm, being a plant used by the Natives of America for the cure of that Disease, which by the Influence of Sir Wm. Johnson he obtamed from the Savages which he terms Lobelia. This plant Dr. Haller says safely and expediciously cures the Venereal Disease, and more so than any mercurial preparation made use of by Europeans for the Cure of that Disorder. If this account be true, as it seems attested by so good a Voucher, I am of the Opinion it may be rendered of great service in the very worst diseases which affect the Human body, and which indeed are a reproach to the Medical Art, such as these, the Leprosy of the Greeks, Elephantiasis, the Canker which carnes off Thousands of Youth, and old obstinate scorbutic cases. As I have a Patient who I greatly value laboring under the Leprosy, Joyned with a Scorbutic Habit, I should ever acknowledge the Favour, if you Introduction of Lobelia Syphilitica | Am. Jour. Pharm. April, 1932 Am, Jour. Pharm. ) April, 1932 } Introduction of Lobelia Syphilitica lli>r I \ I would be so kind as to use your Friendship with Sr. William to gain the knowledge of this Plant, to procure some of it, as also some of the Seeds, and ye knowledge of the place or Soils best adapted to its Growth. . . . The method in which the natives use it, would like- wise be necessary to know, and its Operation, if it operates any other ways than an Alterative. ... "If I may be favourd with such informations, I shall, with his permission, endeavor in the most extensive manner, to render them of publick Utility, and if I may be permitted. Acquaint the Publick, to whose Benevolence and Goodness they are indebted for the Dis- covery, which I am under some advantage to communicate, either by the Royal Society or the CoUedge of Phisitians at Edinburgh." To this appeal Rev. Samuel Johnson seems to have listened sympathetically since in the Library of Congress there appears a letter from him to Sir William Johnson dated at Stratford, Connecti- cut, November 2, 1767. He qualifies Dr. Gale as an expert and leaves his plea with Sir William. *This Dr. Gale is I beheve the most considerable physician in these parts. He had an Academical Education, and was regularly bred to physick by the celebrated Dr. Elliot, of whom, perhaps you may have heard. He is very ingenious and inquisitive, & has a correspondence with the famous Dr. Huxham in England and other members of the Royal Society, & being a friend of mine, I humbly hope you will excuse my thus writing to you at his desire." Sir William's reply, to be seen in the Library of Congress, was promptly penned. He writes from his historic mansion still to be seen by the interested tourist. "Johnson Hall, December 23rd, 1767. Since my last of the ist of this Inst., I was favoured with yours of the 2nd November inclosing (Doctor Gale's) letter to whose In- genuity, Abilities and Character I could not refuse the Satisfaction he recognizes concerning the plant which Dr. Haller considers as Efficacious in The Cure of the Venereal Disorder touch (ing) which I must acknowledge the Truth of Dr. Gale's information. . . . Mr. Kalm an Ingenious Botanist from Sweden, on a tour through this Country for obtaining Usefull Subjects in the Way of his Study in the year (blank space) applied to me for advice & Countenance to Enable him to prosecute his design, which he readily obtained with a Protection & Escorts of Indians that enabled him to go as far as the Great Falls of Niagara, then deemed a bold Undertaking. In a Conversation with this Gentleman on his return concerning the many Medicinal plants, which are used with Great Success by the Indians, 1 took occasion to mention that which is the Subject of this Letter and Gratified his enquiries about it with an Exact Description of the plant & an acct. of its Extraordinary Effects — ^by which means it came to the Knowledge of Dr. Haller, but how far that Eminent Physician has been enabled to describe it I can't tell. I shall there- fore readily give Dr. Gale all the Information I can as well as pro- cure him the plant when the season permits, heartily wishing it may thro' his Means be introduced and be found beneficial to the publick in Cases he Describes. "This plant as near as I can at present recollect has many white Fibrous Roots & grows in the stalk to be about 2 feet in heighth bearing a flower the Cup of which is in the form of a bell, and of a fine Blue Colour. It grows only in cold Swampy Grounds, and is to be found in many parts of this Country particularly on one part of my estate, from whence I furnished Mr. Kalm with the Specimen. The Indians use it both as a Decoction and Lotion. They boil the Roots, the Juice of which they Drink washing the parts Likewise with the Liquid. They also use it with great success in Disorders of the Bowells, but the former I shall fully Describe in the Manner required and also procure them for Dr. Gale as soon as possible in the Spring, for at present we have Two feet of snow hereabouts so that it cannot be had— and this must at present apologize, for the imperfect description I have given of it . . . " The correspondence seen closes with a brief note dated at Strat- ford, March 21, 1768, in which Rev. Samuel Johnson gratefully acknowledges the foregoing communication that Dr. Gale was likewise ^'vastly obliged" to receive. Botanical Laboratory, University of Pennsylvania. •«• Beprinted from Bartonia Special Issue, 1931 John Bartram's Life and Botanical Explorations RODNEY H. TRUE Mr. Chairman : — We meet this afternoon to commemorate the life and achievements of one who was rated **as a plain unlettered man," a farmer living in the colony of Pennsylvania some- thing over two centuries ago. Other men whose names come down to us from those years were his honored friends and acquaintances. We hear of Benjamin Franklin, the philoso- pher ; of James Logan, the proprietary representative in the Colony ; of Dr. Cadwallader Colden, high official in the Colony of New York ; of Dr. Fothergill, famous London physician ; of Sir Hans Sloane, the physician to George II ; of Linnaeus, the great organizer of Natural History; of Peter Collinson, the English cloth merchant doing business in two hemispheres. It is hardly strange that these men should have been remem- bered because they had their hands on the great affairs of their time and places. But what has preserved along with their names, that of *Hhe plain unlettered farmer" from the banks of the Schuylkill? He did two things that made his name known even to our present day; he added greatly to mankind's store of knowledge and— he planted a garden. These results were the outcome of a life of extraordinary physical effort directed by a mind consumed by a curiosity to know and possessed by the instinct to preserve the knowl- edge gained in a lasting record written in living things. There was, in those days of discovery, an eagerness for infor- mation concerning lands only just being realized, this eager- ness springing in part from the possibility that treasures of unguessed value might be found and in part from the disinter- ested urge to know the unknown. It was an age in which alert and expanding minds were eagerly and romantically exploring new worlds. Something like this is now to be seen in the wide-spread interest in the physical world. As we are (7) 8 TWO HUNDREDTH ANNIVERSARY OP now all keenly watching for new developments in the physics of the atom, in the source of cosmic rays and in the structure and extent of the universe, so then electricity was engaging the attention of the kite-flying Franklin ; of Eliot, the farmer- preacher of Connecticut; of Governor James Logan here in Philadelphia ; of Bartram, the farmer-botanist on the Schuyl- kill and of the body of intelligent men. This same eagerness was also fully felt in the mother country where the cloth merchant, Collinson, the Dutch physician-botanist, Gronovius, the Duke of Norfolk, Lord Petre, Queen Ulrica and even King George himself kept their knowledge up to date through con- versation or correspondence with scientists and with others interested in science. John Bartram came rather suddenly into conscious relation with this spirit and was soon in the circle himself. Against such a background, we must agree that his studies stand out as an eminent individual contribution. Let us for a moment trace the familiar story of his ante- cedents. It was believed that the Bartram family in England was of Norman French origin. In time, the religious fermen- tation that drove many Englishmen to seek greater freedom in the New World where the crust of custom and law had not yet hardened sufficiently to restrain a measure of freedom, also drove a group of Derbyshire families to the New World where, following the lead of William Penn, they arrived in 1682, the year in which Philadelphia was founded. The com- pany settled in what is now Darby in the county of Chester. Here they took advantage of their new found liberty, and in the same year formed a meeting at Darby. Among the mem- bers was John Bartram, who had brought with him from England three sons, John, Isaac and William. William mar- ried Elizabeth Hunt of the Darby meeting and they had three sons, John, James and William. John, the eldest son, became the botanist whom we to-day celebrate. He was born in 1699, inherited a farm near Darby from his uncle Isaac, and seems to have been a farmer all his life. Here he had an oppor- tunity to cultivate a taste for natural history that had been FIRST BOTANIC GARDEN IN AMERICA with him even in childhood. He learned the virtues of the plants about him and with this information he seems in his earlier days to have administered to the sick. It is said that he added surgery to medicine and relieved his neighbors who, remote from the city, were in need of skilled help. His inclination toward medicine appears at times in his later writings and observations. W^hile still a young man he underwent an experience that might be likened in its effect on him to that wrought on Saul of Tarsus when the great light shined on him while on his journey to Damascus. The young farmer-physician, resting from his ploughing, observed more carefully than had been his wont, the details of the construction of the flower of a daisy. This aroused a train of meditation that resulted in what we may fairly call a conversion. ''This seeming inspiration suddenly awakened my curiosity, for these were not thoughts to which I had been accustomed. I returned to my plow, but this new desire did not quit my mind." In spite of the prudent caution of Mary Maris, his wife for four years, in spite of his lack of money, he had to ' ' follow the gleam. " * * At last, I could not resist the impulse ; for on the fourth day of the following week, I hired a man to plough for me and went (as many a man in search of wisdom has gone) to Philadelphia/' Here he bought books on botany in a language he could not read. He hired a schoolmaster to teach him Latin and so intent was he on his subject that in three months he could read Linnaeus. Now his fate was sealed.^ He studied the plants of his place and of his neigh- borhood. He soon ranged more widely and began that life of pilgrimage that brought much of the plant life of the eastern part of our country to light and enriched his beloved science as it probably never again can be enriched by any one man. Usually he stayed at home until the early fall saw the harvest over and then began a tour into the unexplored country about him. Sometimes it was toward the Blue Mountains to the iFor a further discussion of '*John Bartram 's first Interest in Botany" see page 35. 10 TWO HUNDREDTH ANNIVERSART FIRST BOTANIC GARDEN IN AMERICA 11 1 ii-i ! iv^- northward, sometimes eastward into Jersey with its weird plant population, sometimes to the south into Maryland. In his travels he often stayed with Friends of other meetings. His traveling began to take a wider swing when he was about thirty years old. He had lost his wife, Mary, and in 1729, after two years, had married Ann Mendenhall of Con- cord Monthly Meeting. Apparently Ann either did not share Mary's objections to the life of exploration or accommodated herself and the family affairs to it. At all events, John traveled farther than before. It seemed desirable to him, as to his tribe since, to bring home something from his travels, something that could witness to the wilds explored. Accord- ingly, he began to bring back and plant seeds and roots of interesting things seen. He needed a place specially dedi- cated to science and beauty, for he found great beauty in many of the new plants seen. Hence on September 30, 1728, he purchased at sheriff's sale a tract of ground that became in time his garden. What another man had not found worth the taxes, or perhaps what another man had been forced un- willingly to give up, became the cherished home of John and Ann Bartram. With his own hands he fashioned out of the stone of the region the house that became his home, probably in 1731. These years from 1728 to 1731 seem to have brought much cause for happiness to Bartram. He again had a helpmate, he had begun his garden and had set his house in it. He had seen treasurers that botanists had never seen before. He had often known that thrill of discovery that comes rarely to the botanist of the present time. But he had fallen out with the Friends at the Darby Meeting and they had rewarded his determined lack of orthodoxy by excommunicating him. He seems to have taken this action in a manner that testified strongly for his sincerity. He continued to go to Meeting and to enjoy the fellowship of Friends, declining to let this official disapproval alter his course of life either in following the inner light that had shone in him or in enjoying the help- ful association with his fellows. That he remained firm in his convictions is shown by the inscription engraved by him on a stone block built into the wall of his new house some years later. "Tis God alone, Almyty Lord, The Holy one, by me adored. John Bartram, 1770 ' ' It Still witnesses to John Bartram 's fundamentally deep religious character and to the steadfastness of his mind. The religious note here sounded appears and reappears in his letters and journals and a real and rugged eloquence breaks forth at times into a sublimity of expression recalling that of the prophets of old. „ , /^ ^ A reverence and wonder at the mighty works of the Creator seen in some of the wildernesses visited by him find their way into descriptions of things seen and into his explanations of how things came to be. John Bartram was a man of faith and the more he saw the deeper grew that faith. Not long after he began to plant his garden his fame had begun to spread among those "of curious mind." Franklin's Junto fellowship begun in 1727 had not included Bartram, but it did include a middle-aged Philadelphia mer- chant of agreeable personality and inquirmg mmd, Joseph Breitnall Franklin describes him as a scrivener but that he had financial dealings with the London cloth merchant, Peter CoUinson, seems certain. Like the other men in Junto, Breit- nall had a mind busy with more than papers and merchandise. He too was a naturalist and as such a brother under the skin to John Bartram. Peter CoUinson in England was a man ot wealth doing a business in cloths both at home and in the Colonies. He was also one of the men "of curious mmd, and likewise a naturalist. He had some inklings of the rare things to be had in America and pestered his American corre- spondents to get plants and seeds for him. They did a little, but far too little, for the enthusiastic CoUinson. Fmally, he took the plain hint from one of his friends who would buy cloths from him but would not be bothered about plants and seeds. So he asks BreitnaU, Dr. Samuel Chew and perhaps 12 TWO HUNDREDTH ANNIVERSARY OP FIRST BOTANIC GARDEN IN AMERICA 13 t other Philadelphians to give him the name of some one who would serve his purpose. Both recommended Bartram and thus began the lifelong connection that soon became not only a business relation but one of close friendship. CoUinson himself was a Friend, like Bartram, given to plain and some- times emphatic statement. In the thirty-eight years during which they exchanged seeds, plants, letters and plain speech, relations were sometimes momentarily disturbed by the great eagerness of CoUinson who little appreciated the herculean labors performed by Bartram in getting the desired plants for him and by John's reproaches at his lack of appreciation. And, as the later years came on bringing the crisis of the Eevolution, Peter could not sympathize very deeply with the colonies and John regretfully drew back from the mother country. But although Peter could see an excuse for mas- sacres by the Indians in the wrongs they had suffered at the hands of the whites, John in a quite un-Friend-like attitude said that the only remedy was to **bang them a plenty.'' Again John was unorthodox. But he and Peter never allowed the deeper current of their friendship to be disturbed. As the interchange of plants, seeds and opinions between Bartram and CoUinson became known and fruitful, other con- nections were made by Bartram with Europeans wishing either to enlarge their collection of rarities, or with men of science seeking knowledge of the New World. Sir Hans Sloane, Royal physician to George II, whose huge collections later became a basis of the British Museum, was one. Dr. Pothergill, a wealthy London physician, and later patron of William Bartram 's travels to Florida, was another. Among the correspondents most flattering to Bartram 's reputation was Queen Ulrica of Sweden. He was also solicited by the leading naturalists of Europe. The name that has grown with time, that of Linnaeus, the Swedish systematist, is among those who valued Bartram and who received his plants and his let- ters. The Dutch Gronovius wrote Bartram letters in such curious terms that Bartram begged him to use a language that he could understand. In his own country he became the botanical ** central' ' car- rying on a correspondence with Colden in New York, Dr. John Mitchell and John Clayton in Virginia, Jared Eliot in Con- necticut and Dr. Garden in Carolina. Ellis, Philip Miller of the Gardener's Dictionary, James Gordon, Mons. Dalibard of France, Martha Logan, **the fascinating widow" of Charleston and author of the first American book on gardening. Colonel, later General, Henry Bouquet of Fort Duquesne fame, Thomas LamboU and Henry Laurens are among the well known people of his time with whom he corresponded. He traded ideas often and abundantly at shorter range with Ben- jamin Franklin. Dr. Dillenius of the University of Oxford interested Bar- tram in mosses. John writes to Mark Catesby, the author of the elaborate and gorgeous volumes on **the Natural History of Carolina, Florida and the Bahama Islands"— ** Before Dr. Dillenius gave me a hint of it, I took no particular notice of Mosses, but looked upon them as a cow looks at a pair of new barn doors." His interest in mosses eventually received recognition when a beautiful American genus was named Bartramia to commemorate his name. It is hardly to be wondered at, therefore, that the plain unlettered plowman impeUed by the consuming loyalty that grew out of the contemplation of the daisy should receive open and flattering recognition from scientific organizations even before the end of his long life. Through the friendly CoUinson 's influence several of his contributions found their way to the Royal Society in England and were there printed. When the American colonies, Pennsylvania taking the lead, with Franklin as prime mover, established its own American PhHosophical Society, it was to be expected that Bartram, by now recognized as the first American botanist, should be a member. On April 5, 1744, a society was formed with about a dozen constituting members, Bartram being one. In the remaining minutes aUotted to me, I shall discuss briefly the travels of John Bartram. It must be borne m mind that well established and comfortable roads in his time Ill t4 'f 14 TWO HUNDREDTH ANNIVERSARY OF FIRST BOTANIC GARDEN IN AMERICA 15 were to be found mainly near larger centers of population and ran out from these into sparsely settled, remoter regions. These roads frequently amounted to little more than well- marked trails, in many cases following those used by the Indians. When these roads ceased, the traveler was obliged either to continue along less marked Indian trails or to enter the untracked wilderness. To the botanist, the object sought was not primarily a route of travel from one place to another but rather access to regions in which rare or interesting plants might be found. Accordingly, in trying to make clear to friend Peter CoUinson some of the problems of the collector, he says that his route led him usually into regions in which travel was difficult and often dangerous. He writes in the same vein to Alexander Catcot in 1742, *Hhee may suppose I am often exposed to solitary and difficult travelling, beyond our inhabitants, and often under dangerous circumstances, in passing over rivers, climbing over mountains and precipices amongst the rattlesnakes, and often obliged to follow the track or path of wild beasts for my guide through these desolate and gl oomy thickets. ' ' A glance at Lewis Evans' map of the Middle Atlantic region in 1749 shows that roads to the northward stopped at the Kittatiny Mountains or clung to the Delaware River. New Jersey being nearby and less rugged was naturally more accessible and was often traversed by our botanist. One road into Delaware led to the Atlantic Coast at Lewes, another to the shore of Chesapeake Bay. Roads to the westward were more adequate because of the considerable settlements in that direction, as Lancaster and York. These parts of the country were easily reached and were probably repeatedly studied. He made several longer trips, however, that deserve a passing mention. On these trips he made brief notes on the character of the landscape, principal kinds of forest trees seen and geo- logical features with special mention when fossil outcrops were noted. He included meetings and doings with the Indians and I believe rarely failed to mention the seeing of a rattlesnake. Sometimes his journal contains amplifications, explanations of possible causes of phenomena seen. He seems to have sent these journals to his European correspondents, especially to Peter CoUinson. In two instances, Peter CoUin- son got them printed in England, but by far the greater number seem to have remained in manuscript. It is to be hoped that these still exist and that they may sometime be carefully examined by some one competent to judge their value for publication even now two hundred years later. To follow the always-wandering Bartram can not be at- tempted at this time. Three of his journeys however had some wider significance and have been rather clearly traced. Some idea of the restless activity of this botanist, who was also running a farm, may be gained from his correspondence. In 1735, at CoUinson 's suggestion, he foUowed the Schuyl- kUl River to its source and in 1737 he finished the preparation of a map of the region traversed. In 1736 he visited the Rattlesnake Mountains and went into the Jerseys. In 1737 he went to the westward to Conestoga and planned his first longer trip, this time to the southward. He set out in the f aU of 1737 apparently going down through Delaware and the Eastern Shore sections of Maryland and Virginia, thence up the James River by way of WiUiamsburg to the mountains, turning northward through the Shenandoah Valley. On this trip he reports seeing a cave so remarkable that he *ook the pains to prepare a plan of it. On this expedition he traveled 1,100 miles in five weeks. Travels up the Delaware to Minne- sink on the west side of that river on the line between West Jersey and New York, iris-like flower from Cape May, balsam from firs gathered in the CatskiU mountains, and an indefinite journey probably not of great length ''along our sea coast'' are not noted in 1742. In July 1743 in company with the interpreter to the Indians, Conrad Weiser, he went on one of his most important journeys, this time beyond the Blue Mountains across a region little known to the Onandago country near the present site of Syracuse, N. Y., where a meeting was to take place between Indians of the Six Nations and representatives of the colony 16 TWO HUNDREDTH ANNIVERSARY OP of Virginia. There had been trouble and some killing and all parties desired peace. John went along in order to make observations on the plants, animals and physical characteristics of this wild inland region. With the help of the early maps of Lewis Evans and William Scull it is possible to follow his journey fairly well. We may take a minute and follow him a little more closely on this one journey. Leaving Darby on July 3 he crossed the Schuylkill to Philadelphia and, probably taking the Ridge Road, came to the Perkiomen Creek near its junction with the Schuylkill. He spent the night at Marcus Hulin's in Mana- tony south of the Oley Hills. On the Fourth he crossed the Schuylkill, perhaps near where Pottstown now stands to the west bank instead of following the usual trail on the eastern side to the present site of Reading. He ascended the Flying Hills, south and west of Reading, so called, he remarks, be- cause it was the home of great numbers of wild turkeys that were seen to fly down from thence into the valleys. This brought him to near Reading and the high hills in the neigh- borhood of Wernersville. He saw the Blue Mountains beyond the Tulpehocken Vale, passed a strikingly large spring, near the Sinking Spring of to-day, and, turning westward along the road that soon ended in the wilderness, reached the vicin- ity of Womelsdorf, the home of Conrad Weiser, his compan- ion from this point onward. After spending the night at Weiser 's, the two crossed Tulpehocken Creek where, accord- ing to Evans' map of 1749, the road ended and they set their faces toward the Blue Mountains. They crossed the Little Swatara and spent the night of the fifth at William Parson's plantation six miles from the Blue Mountains. The next day saw them through the first range probably northwest of Rehrersburg in the town of Bethel and showed them two gaps in the second range. They probably crossed this mountain at Pine Grove, passing over the principal branch of the Swatara and traversed intervening swamps, hollows and small ridges, perhaps *'St. Anthony's Wilderness," of the maps that time, to the last great ridge. They spent the night on FIRST BOTANIC GARDEN IN AMERICA 17 Laurel Creek, perhaps Deep Creek of present day. One more ridge crossed and they came to Double Eagle, now Spread Eagle Creek, going steadily northward. Here they turned westward and from the top of a high hill, with shells in the rocks, saw the Susquehanna. They spent the night at Ma- honey. On the 8th they crossed the creek and went north- ward along the eastern bank of the Susquehanna that led them to the junction of the West and East Branches of that river to an important Indian town then called Shamokin, later Fort Augusta, now Sunbury. A day was spent in re- connoitering this region, the tenth seeing them again bound northward along the western branch of the Susquehanna to the great westward bend where Muncy Creek joins it. They soon held northward to Lycoming Creek and westward to Bur- nett's Hills, and, leaving the ** Impenetrable Wilderness" with the ** Endless Mountains" of the Eaglesmere region still farther to the eastward, reached the watershed separating the waters of the east and west branches of the Susquehanna. Crossing this they turned eastward to Towanda Creek. It was now July 16 and they were on water that flowed north- ward at Owego on the present state boundary line. It seems probable that here their course led north by the valleys of East Creek and Tioughnioga River past the present site of Cortland, N. Y., perhaps to TuUy Lake, then over the Goose- berry Mountains. They made their camp on July 20 near their destination. Indians, who had joined them at Shamo- kin, like them en route to the council, were sent ahead to the place of rendezvous with wampum and word that they were coming. Next day Table Mountain with its fruits and vege- tables was crossed, then water flowed toward Lake Ontario, and the Onondago long house somewhat south of where Syra- cuse now stands came into view on July 31. To Bartram's great regret the return trip nearly retraced this route, but before he returned Bartram took a side trip to Fort Oswego on Lake Erie while the treaty was making at Onandago. A brief journal, such as he usually prepared recounting his travels, was sent to his friend CoUinson in England who had 18 TWO HUNDREDTH ANNIVERSARY it printed wretchedly in 1751. This journal notes hard climbs, dismal vales, glorious far views, the prevailing forest trees, types of soil, doings with the Indians and, wonderfully enough, marine shells in the rocks on high mountains. Some of the difficulty of corresponding in those times of uncertain travel and of wars in progress on sea and on land is realized when John complains that he prepared and sent three copies of this journal before one reached England. The two earlier copies went in boats picked up by French priva- teers. Time forbids more than the briefest mention of other trav- els. In 1753, with his son, William, now fifteen years old, to keep him company, he ascended the Delaware to the vicinity of Milford or Port Jervis, then across to Goshen in New York and up toward the Hudson to Cadwallader Golden 's place west of Newburgh. In 1760 one learns of a trip to Gharleston, S. G., in which he met Mrs. Martha Logan, the aged and vigorous author of the first American book on Gardening. In 1761, after the English had taken Fort Duquesne and christened the place Pittsburgh, he visited it and received from his lasting friend Golonel Henry Bouquet, the hero of the situation, the first pecans that found their way into the east. With these he greatly aroused the curiosity of his English friends, Gollinson and Gordon. Next year finds him on a long trip that brought him into the interior of South Carolina, to the southwest border of Vir- ginia, to the New River and through the Shenandoah Valley in the direction of home. He saw the famous Natural Bridge, Luray Gaverns perhaps. Hot Springs and other won- ders that stirred him to a real eloquence. This trip yielded an interesting crop of letters and a journal with maps never pi inted. 1763 finds him near home visiting Great and Little Egg Harbor but planning a long trip to Florida. England had just assumed charge of this new colony and it was agreed that it needed investigation. Through the intervention of FIRST BOTANIC GARDEN IN AMERICA 19 Peter Gollinson, John Bartram was appointed Botanist to the King in April, 1765, with a modest money allowance and authority to proceed. By September 19, he and William were safely on their way as far as Garolina. Georgia and Florida were visited and a journal of travels to the head of the St. John's River was published in several editions under the chaperonage of Dr. William Stork, a man greatly interested in the first land boom experienced by that since much-boomed state. This was Bartram 's last long trip. He was sixty-six years old and was beginning to feel the weight of years. He had suffered much hardship and the help of his boy John as well as that of the rather difficult but talented William was gladly accepted. As the Revolution came on, the country was much disturbed and John complained bitterly and often of the dangers from savages let loose. So he traveled less, probably giving more time to his garden that had now taken on somewhat the char- acter of a nursery for the production of plants for the many customers at home and abroad. His garden, speaking to him of hardship and not rarely of peril, of triumphant discovery, and of peace, was to him far more than his fertile farm acres that quietly and beneficently had yielded their prosy increase through the years. As the struggle between the mother country and the colo- nies wore on, war came to his very garden, to his very door. It is said that Howe's progress up the river toward Philadel- phia troubled him greatly and as Howe entered, our old bota- nist started on his last long journey. Much might be said on this occasion about the life of Bar- tram. But, when it is all summed up, we find that we have had to do with a simple-minded but far-seeing, beauty-loving apostle of Nature — all his life a zealous convert with a mission humbly but greatly conceived and executed with all the pow- ers that he possessed. He wrote to Gollinson — *'My head runs all upon the works of God, in Nature. It is through that telescope I see God in his glory." Reprinted from Bartonia, No. 13, 1931. Kyllinga pumila in Philadelphia Rodney H. True Late in September, Mr. James Lambert, Superintendent of the Botanic Garden of the University of Pennsylvania, brought to my attention a plant that had appeared abundantly on a piece of moist ground in the Garden near the pond. A careful comparison with material in the Herbarium of the University showed that it was Kyllinga pumila Michx. far beyond any range there seen. This determination was con- firmed by my colleague, Dr. John Milton Fogg, Jr. The standard manuals give the northern range as Maryland, Ohio, Illinois, with Virginia and Delaware cited in some cases as the farthest point to the north in the eastern part of the country. An inspection of material in the Herbarium of the Philadelphia Academy of Natural Sciences showed a speci- men collected in Sussex County, southern Delaware, by Com- mons in 1875. Collections were seen from North Carolina, South Carolina and Georgia, and from as far north as Ohio and as far west as Iowa and southward into Kentucky and Tennessee. It seems clear, then, that the abundant growth on an area perhaps 6 ft. by 12 ft. represents an occurrence well outside of its normal range. One asks himself how this may have come about, and the first suggestion, in the light of Darwin's investigations on the muddy feet of migrant water fowl, is that some water bird coming from the south may have alighted in the Garden on the moist soil near the pond and left these seeds of Kyllinga picked up in some similar situation to the southward. Since these plants seeded profusely, it will be a matter of interest to see whether the species reappears next year. Botanical Laboratory, University of Pennsylvania. (47) III' II UNUSUAL ASPECTS OF MEIOTIC AND POST- MEIOTIC CHROMOSOMES OF GASTERIA HSU-CHUAN TUAN Reprinted for private circulation from The Botanical Gazette, Vol. XCII, No. i, September 1931 PRINTED IN THE U.S.A. II.- n UNUSUAL ASPECTS OF MEIOTIC AND POSTMEIOTIC CHROMOSOMES OF GASTERIA^ Hsu-Chuan Tuan' (with plates I, II AND FOURTEEN FIGURES) Introduction The investigation here described was initiated at the University of Pennsylvania in 1929. The preHminary work consisted of a study of the comparative action of different fixing reagents and of the methods of applying different nuclear stains. The investigation cov- ered a wide range of plant material. The technique of making cytological smears (20) rendered the question of fixation comparatively simple. Emphasis was therefore placed upon the acquiring of a satisfactory staining technique. It was found that picric acid destains material which has previously been treated with haematoxylin and safranin, and by its use the writer was able to get consistently satisfactory results. Gasteria was first investigated, and subsequently smears of Clarkia, Solanum, Pinus, and many others were studied experimentally. In the course of this study, a group of Gasteria smears, showing microsporogenesis, yielded an unusual phase of diakinesis chromosomes. This discovery led to the present report. In diakinesis, some cells showed a sudden change in their capacity for absorbing the stains, and in addition they were found to go through a regressive process to a false resting stage. Often the pollen mother cells of this particular plant divide three successive times, forming eight pollen grains. Taylor (21, 23, 26) had previously made a thorough study of Gasteria chromosomes, and contributed much to our knowledge of their shape and structure. A comparable investigation of this series of abnormal chromosomes in the same genus might therefore be expected to reveal other interesting facts. X Contribution from the botanical laboratory of the University of Pennsylvania. ^ Fellow of the Chinese Educational Mission at Washington, D.C., of Tsing Hua University, Peiping, China. , [Botanical Gazette, vol. 92 46 BOTANICAL GAZETTE [SEPTEMBER A part of this work was done at the Marine Biological Laboratory, Woods Hole, in the summer of 1930, after the material had been collected and partially studied at the University of Pennsylvania. The research was then completed at the botanical laboratory of the University of Michigan. To these institutions the writer is much indebted for laboratory space and the necessary equipment. Material and method Pollen mother cells of Gasteria were collected in a greenhouse at the University of Pennsylvania in the spring of 1930. Young buds were teased out with a needle and the anthers transferred immedi- ately to a thoroughly cleansed slide. With a stroke of a scalpel, the pollen mother cells were smeared evenly into a thin fihn on the slide, which was then submerged in the fixing fluid for 20 minutes. Many fixing fluids were tried, and although the results varied much in detail, the fixation of smears was generally better than prepara- tions sectioned by the parafhn method. The following account is based upon the result obtained by the use of a special chromic-acetic- osmic fixative developed by Taylor (McClung ii) for Gasteria pollen mother cells. No paraffin preparations were used, since the smears give the most natural image of the cell, both in regard to its general contour and to the details of its chromosomes. Shrinkage is almost eliminated in most of the cells. The staining technique described by Tuan (27, 28) was followed, with some slight modifica- tion to suit each individual case. When haematoxylin was used, the picric acid differentiation was occasionally abbreviated by the sUdes being placed in a 0.5 per cent iron alum solution for 2 minutes before they were transferred to the picric acid for further differentiation. This saves the long schedule of differentiation required when picric acid alone is used. For Gasteria, however, the writer still prefers the long schedule of picric acid, but in the case of Tradescantia, prepared for purposes of comparison, a previous treatment in a 0.5 per cent iron alum solution cut down the time from 3 hours to 30 minutes. When stained with safranin, the slide was brought into a mixture of two parts of clove oil and one part of 95 per cent alcohol, and there the differentiation was further adjusted. Deeply stained or over stained sUdes can be saved by this process.^ One example 3 This alcohol-clove oil differentiation can also be applied to crystal violet smears 1931] TUAN— GASTERIA CHROMOSOMES 47 from each of the methods illustrates their general application to the Gasteria study: Slide PK 8, Safranin Minutes 1. Stained in Grubler's safranin O 5 2. Picric acid-alcohols 5 3. Ammonia-9S% alcohol 0.5 4. Pure 95% alcohol 0.5 5. 95% alcohol-clove oil mixture 5-10 Slide KP 9, Haematoxylin^ 1 . 2% iron alum 20 2. i% haematoxylin S 3. Differentiated in. picric acid 160 Slide KP 6, Haematoxylin 1. 2% iron alum 20 2. i% haematoxylin 5 3. ^% iron alum 2 4. Differentiated in picric acid 30 Investigation In the normal behavior of Gasteria chromosomes, according to Taylor (21, 23, 26), the first suggestion of prophase activity in meiosis is marked by an aggregation of chromatic granules, forming more or less spiral chromonemata. When the spirals straighten out from a tangled condition to form the extended chromatic threads, synapsis of homologous chromosome pairs occurs simultaneously at different places throughout the nucleus. The longitudinal split of the paired chromosomes, which form the tetrad, is accomplished before the diakinesis stage. Three pairs of pyriform chromosomes and four pairs of larger chromosomes are present on the metaphase plate. The tetrad condition of each pair is revealed distinctly. A coiled spiral of from seven to nine turns is present in the longer chromosomes, while in the small ones the number of coils is cor- respondingly reduced. These coils apparently are a result of the contraction of the more or less straight chromonemata of an earlier stage. In the anaphase, the two chromatids of each dyad may sepa- rate and the coils within them become more distinct. When telo- phase is reached, the chromosomal sheath is no longer stainable but 4 When haematoxylin is used, each stage is followed by a prolonged washing in tap water. V, 48 BOTANICAL GAZETTE [SEPTEMBER is Still present. Eventually the chromosomes become disorganized in the interphase. When they reappear for the second division, their shape and internal structure acquire interest from a new angle. At the proximal end of each chromosome a mild constriction is invari- ably present; perhaps in the three short ones a secondary constric- tion occurs along the shaft. In size, the homeotypic chromosomes are more slender and longer than the heterotypic ones. The internal structure of each of the homeotypic chromosomes resembles that of the somatic chromosomes, that is, each chromosome contains two chromonemata derived from a split of the coils. A quartet is formed as a result of two cell divisions. Each of the resulting cells divides again after the formation of a thick wall, typical of pollen grains. There are several striking differences between the normal behavior of Gasteria chromosomes, as just described, and the peculiar mode of division to be recorded in the next few paragraphs. The first sign of abnormality is a dispersal of the diakinesis chro- mosomes to form a false resting stage. More complicated abnor- malities occur after the first metaphase. The nuclear membrane fails to form at telophase. Two sets of second metaphase chromo- somes persist through the omission of the interphase and second prophase. The chromosomes thus produced assume two different aspects. The type (fig. 30) with distinct spiral structure is identical with that which appeared in the first metaphase; the other type (figs. 2, 8, 10, 18) is erratic and cannot be connected with any par- ticular division of the chromosomes. So far as their outer form is concerned, they are intermediate between the homeotypic and heterotypic chromosomes, as they have extraordinary terminal heads and short, heavy bodies. Abnormal chromosomes of another type (fig. 9) are produced by some cells in which a nuclear membrane is formed after the telophase. The probable fate of these last was not determined, owing to a lack of sufficient material of this type, but the history of the chromosomes with spiral structure is traced to the formation of third division chromosomes within the nucleus of quartet cells (fig. 7). No regular division of the quartet was ob- served, but furrows of various depths occur in these cells, and have been noted in many preparations. This suggests that the third di- vision is carried through by a simple process of furrowing. On the !IH»"-M»*- >ft m-'M III ^ 6 «i 10 11 Figs i-u —Fig. i, selected chromosome sets showing four types of chromosomes: A first metaphase chromosomes; B, second metaphase chromosomes with extraordinary heads; C, second metaphase chromosomes with shape and appearance of first metaphase chromosomes retained; D, set of constricted chromosomes Fig. 2, se^^^J^ metaphase figure showing giant headed chromosomes scattered irregularly m cytoplasm 28 ele- ments counted in all, but only 26 shown. Fig. 3, pair of anaphase chromosomes showing Kng matrix Fig 4, early telophase showing partial rotation of daughter groups. Rg% l^of ^^^^^^^ chromosomes from binucleate cell showing satellite at lower end Fig. 6, mid-anaphase figure showing partial rotation of daughter groups and non- seDaratbn of single chromatid. Fig. 7, third division chromosomes with all types of foS'onf reSsented. Fig. 8, gUp of second -taphase chromo^^^^^^^^ giant headed chromosomes and extruded dyad. Fig. 9, constncted ^h^^"^^^^^^^,^ ^^ binucleate cell. Fig. 10, two sets of giant headed chroniosomes show^^^^^^ 'staS Fi« "^i 2 atDerioherv Fig n, extrusion of small chromosome at equatorial plate stage, i^ig- 12, IcS poYys J!fe. Fig. 13, second metaphase figure showing extrusion of dyad from each of two sets of chromosomes. Fig. 14, Q-celled polyspore. 50 BOTANICAL GAZETTE [SEPTEMBER I931] TUAN— GASTERIA CHROMOSOMES SI other hand, these chromosomes, like some of the diakinesis elements, may disintegrate and produce false resting nuclei. Regression of chromosomes from diakinesis The clearest evidence of tetrad chromosomes was observed during the diakinesis stage (fig. 16). From the general appearance of these threads, it seemed that the cell was perfectly normal, and had be- haved as described by Taylor. The only peculiarity up to the first metaphase was a suggestion of disintegration of the thickened dia- kinetic chromosomes, which were recognized in a few cells by the fact that they stained faintly (fig. i?)- When these chromosomes are traced through several stages, it is found that they collapse com- pletely. The tetrad is gradually brought clearly into view with the homologous chromatids connected by numerous achromatic bridges (fig 32) The chromosomes may be compared to a rope ladder. In a later stage the achromatic bridges are replaced by a clear space so that parallel threads of an early prophase nature are formed (fig 29) Further disintegration of the threads brings about a false resting stage, which might be mistaken for the earliest evidence of the first prophase, except that the nucleolus in this case is absent. Probably cells of this type will not divide again, as there was no evidence to show a reorganization for further division. Those cells which show the same kind of disintegration but at a later stage will not be described in detail. A brief account of their development, however, may bring out some points for discussion, (i) Empty cells are frequently observed among all stages of meiotic figures. (2) Cells at the binucleate stage or quartet stage which show only a large but faintly stained nucleus are also frequent. (3) Nuclei of a quartet may show different stages of activity (fig. 40), sometimes from one to three nuclei of a quartet being missing. Whether these cells represent a complete breakdown of the chroma- tin material or not could not be determined, but from this example of the diakinetic chromosomes it seems likely. As to the cause of these abnormalities, it is rather rash to connect them with any one agent, such as pathological influence or genetical unbalance. It is most probable that there were other forces which, in addition to these, contributed much to the peculiar behavior of these cells. First metaphase. — The lapse of time from diakinesis to meta- phase is exceedingly short. This observation is confirmed by a gen- eral lack of intermediate stages in many preparations, although in a few cases figures of such were not hard to identify. When diakinesis ends, the chromosomes shorten and thicken with surprising rapidity. The scattered chromosomes are suddenly gathered up in the central portion of the cell, while the nuclear membrane disappears entirely and the fiber attachments are formed on each of the chromosomes. It seems that the chromosomes which are now not in contact with the cytoplasm are able to take up the stains more readily than at any other stage ; therefore the metaphase chromosomes arranged on the equator have the appearance of solid rods, and their internal structure is detected only in well differentiated preparations. On the metaphase plate, seven pairs of chromosomes, spUt into tetrads (fig. 27), can be observed so far as the general contour of the cell and the chromosomes are concerned. There are, however, several strik- ing cases of abnormal distribution. In a few cells one of the small chromosomes is found at the polar region when the rest of the set is still on the equator (fig. 11). Also, one of the small chromosomes is often extruded entirely from the spindle region (fig. 6), so that when migration is completed, it is left at the peripheral region of the equator where it rounds up quickly to form a small nucleus. First anaphase. — The anaphase chromosomes can clearly be counted with seven dyad chromosomes in each daughter set. At the beginning of migration, the chromatids are evidently subjected to a strong pull between the spindle fibers and the surfaces of contact with their corresponding homologues. When they are midway be- tween the pole and the equator, they are stretched to a certain ex- tent, the turns of the spiral bulging out at places and suggesting the presence of an elastic matrix (fig. 3)- Slender threads may be seen between them as the daughter chromatids are drawn farther and farther apart. As a rule, the partners of each dyad are separated before they reach the pole (fig. 31). In one extreme case (fig. 6) one of the chromatids was pulled across the equator and failed to separate. The telophase (fig. 4) follows, with a general lack of the compactness of the chromosome such as is noticed in the normal condition. The smaU chromosomes reach the pole ahead of the large 52 BOTANICAL GAZETTE [SEPTEMBER I931I TUAN— GASTERIA CHROMOSOMES 53 ones, although migration started simultaneously in all. Many cells now pass quickly to a second division without any of the phenomena associated with the heterotypic telophase. Only the least abnormal types, those which retain some evidence of telophase dispersal, will next be considered. However, a brief partial telophase may result in the formation of chromosomes as seen in figs. 2, 8, 10, and this will be described in connection with the second metaphase chromosomes. Residual first telophase and derived chromosomes The ends of the chromosomes toward the polar region approach each other, but the distal ends, extending toward the equator, alter their position very little. This is soon followed by a gradual rotation of the two groups of chromosomes in opposite directions (figs. 4, 6). After a complete turn of 90° they lose their staining abiHty. There are a few examples of nuclear membrane formation, but the majority of the cells do not show such a tendency. The former type of nuclei lead to another abnormal division, while the latter (fig. 24) soon re- cover their staining properties, so that they are seen as full sized chromosomes on the second metaphase plate (fig. 30). The inter- phase and second prophase of the latter type of nuclear division are entirely skipped, and consequently the appearance and structure of the chromosomes are unchanged from the first division, a condition similar to that of some animals in which the interphase and prophase are skipped during maturation divisions of the egg nucleus after entrance of the sperm. In those cells in which the nuclear membrane plays a part, the chromosomes show a progressive loss of staining ability. The mem- brane appears first at one side, usually the side near the polar region (fig. 21), whence it grows about the mass to complete its forma- tion (fig. 19). At this stage, as the nucleus increases in size, the chromatids are so lengthened that a continuous but loosely coiled spiral is seen in each (figs. 19, 21). Further stretching of the spirals (fig. 20 a, 6, c) gives rise to an isolation of a single loop at the proximal end of each chromosome. Later the nuclear membrane, perhaps as a result of the excess lengthening of the chromosomes, is ill-defined and suggests that it is in the process of disintegration. Further evidence derived from the formation of heavily constricted chromosomes (fig. 9) would seem to indicate that the nuclear mem- brane is finally and entirely disorganized. The cause of such a phe- nomenon was not determined, but it is interesting to note that immediately after disappearance of the membrane, two sets of con- stricted chromosomes are found in the cell. The origin of this type of chromosomes is probably that as soon as isolation of the tips of the spirals is accomplished (figs. 20, 23), anastomoses appear (figs. 37, 38). At first they are difficult to recognize, but later they assume a distinct threadlike form (fig. 9). This process is accompanied by the vacuolation characteristic of telophase chromosomes. The nucle- ar membrane is indefinite, and its complete collapse initiates re- organization of the chromosomes. A count of the elements in each of the cells reveals fourteen as the number (fig. 23). Two of them bear extremely big heads, which are further divided by another contraction (fig. 9). Six of them show comparatively small heads, while each of the six small chromosomes is terminated by a round head and has also a median constriction. All of these heads can be mistaken for satellites, as they are connected to the shafts by very slender threads. In one case, one of the large chromosomes showed a small satellite at the distal end (fig. 5). Presence of true satellites in meiotic chromosomes has not yet been reported, and their purely vegetative nature was emphasized by Taylor (23). No inference can be drawn from this sole exception. Second metaphase. — It has already been mentioned that the interphase and prophase are skipped in many of the cells, owing to failure of the nuclear membrane to appear. This account of the behavior of these cells will therefore begin with the metaphase of the second division. The chromosomes which retain the form they had in first division telophase have now recovered their staining ability in a surprising manner (fig. 24), each of the two sets of metaphase chromosomes being arranged on a second equator for the following division (fig. 30). The plane of this division is perpendicular to that of the first, as is shown by the rotation of the daughter groups in the earlier stages. From these they can clearly be seen as two sets, not ar- ranged in a definite pattern. No case could be found in which the chromosomes were not widely separated from one another; this is m\ 54 BOTANICAL GAZETTE [SEPTEMBER 1 931] TUAN— GASTERIA CHROMOSOMES 55 perhaps one of the reasons that a lagging and an extrusion of chro- mosomes will later occur. Rejection of a dyad chromosome was fre- quent while the elimination of a dyad or two (fig. 13) from each daughter set of chromosomes was common. Those eliminated may give rise to some supernumerary spores. Another type of abnormality is found at the second metaphase plate Although many chromosomes show an internal spiral struc- ture there are some cells whose chromosomes show a striking differ- ence (figs 2,8,10,18). These latter are constricted, with extraordi- nary heads at the tip, while the body is of a peculiar type, short and heavy By an analysis of all the figures available, the origm of these chromosomes was traced back to the first telophase, when the nu- clear membrane was just about to form. Consequently it is most probable that at telophase the nuclear membrane of these cells has a very brief existence, so that the chromosomes are only partially disorganized for the homeotypic division. Further collapse of the spindle would undoubtedly give rise to these scattered chromosomes (fig 2) In fig 2 the total count of the chromosomes IS 28, but only 26 can be shown in the drawing. The count of 28 is exactly four times the haploid number. From the size of the heads, it is further determined that the total complex represents four sets of chromo- somes, and each set (fig. i b) has the haploid number of seven. For these chromosomes at least, it shows that the chromatids of each tetrad are completely separated at late anaphase. Identified by the heads, these chromosomes are traced to the resting quartet stage. Late anaphase cells are frequently seen with four sets of chromosomes spread widely in the cytoplasm (fig. 26). Disintegra- tion of the chromosomes, typical of telophase activity, occurs. For- mation of a nuclear membrane is postponed until disintegration of the chromosomes is almost completed (fig. 33 «-«)• Second anaphase.— The second early anaphase is brief. Cells are often observed in late anaphase (fig. 25) and telophase. It has been stated that the metaphase chromosomes are loosely scattered along the equatorial region, therefore it is to be expected that various abnormalities of the chromosomes would appear in the second ana- phase Several cases of chromosome extrusion from the spmdle re- gions have been mentioned in connection with the metaphase chro- mosomes, but as the appearance and history are similar to what has been described, a repetition will not be attempted. Second telophase and third division chromosomes In many cells, when the chromosomes have reached their respec- tive poles, they round up to form the telophase. Ultimately a regu- lar resting quartet is formed, all the chromosomes having lost their individuality within the nuclear membrane. Probably the meiotic activity of the cells developing in this manner is thereby concluded. There are a number of quartets, however, which retain the spirals throughout the telophase in perfect shape until a new set of active chromosomes is formed (fig. 7). Although these chromosomes were not observed to go through an elaborate division, their formation in the quartet nucleus is sufficient evidence for such a possibility. The history of the spirals here seems to be of the same nature as that shown in the first telophase, although the number in this case is reduced one-half. Furthermore, the constricted chromosomes formed after second telophase (fig. 9) are identical with those found after the first telophase. The development of these chromosomes could be only partially traced. It may be said, however, that at the end of the second telophase, the chromosomes gradually lost their chromaticity as the coiled spiral reappeared (figs. 28, 39). Each of the spirals then elongates at the expense of its width, so that the coils become very narrow compared with those first involved in this process. A loop or two at the tip of each spiral is then gradually dragged apart from the main body as the chromosomes reach their maximum length. The chromosome number is seven: one bears a big head (fig. i d), three are terminated by smaller heads, while at the tip of the small chromosomes a round head is shown with a secondary constriction along the shaft. This combination is exactly like that found after the first telophase, except that each type is represented by a single element. There is not much detail to be seen in these chromosomes. Their structure was discerned, however, in other cells in which the chromosomes were undergoing a process of regression. They had a single spiral. These chromosomes resem- ble the homeotypic ones in their constrictions, but they are different in structure, as the homeotypic chromosome.^ tmder normal condi- i i S6 BOTANICAL GAZETTE [SEPTEMBER tions have two spirals in each. The question arises as to whether or not this type of abnormality should be connected with the normal homeotypic chromosomes. The writer will develop this point later. Several interesting figures may be observed in the regression quartets. The quartet nuclei in many cases show different stages of activity in the same group (fig. 40 a-c). A typical resting nucleus is shown in b; c is shown with partially disintegrated chromosomes; while a is intermediate. From the equal sized and equally stained nucleoli in these cells, it is probable that these stages are passed through in a short period, and the probable disturbance occurs im- mediately after formation of the third division chromosomes. Re- formation of the nuclear membrane later will mark the beginning of the regression process. The most probable steps of chromosome be- havior in these cells are as shown in fig. 22 d, c, b, a, and figs. 36, 40. The earliest stage of disintegration is marked by a loss of chro- maticity of the chromosome matrix, so that the comparatively heavily stained spiral within the chromosomes is brought out in better contrast (fig. 22 c). The chromosome at the lower right hand corner of fig. 36, and two other segments in the same cell, show the spiral clearly, while the farther ends of these chromosomes are in a more advanced stage of disintegration. Increasing number of anas- tomoses and central vacuolation (fig. 22 a, b) bring the chromosomes close to a resting stage (fig. 40 b). It seems that the chromaticity of each spiral varies at regular intervals, and usually certain seg- ments are still heavily stainable while others register little affinity for the stain (fig. 40 a), so nodules are distinctly shown at places throughout the nucleus. When vacuoUzation reaches a climax, the chromosomes appear double, with faintly stained peripheral regions and clear central cylinder (fig. 40 b). Besides these regression fig- ures, there are two selected from a number of quartet cells worth mentioning. The one (fig. 34) with short spirals is rather difficult to assign to a particular developmental position. It is probably a second telophase figure, however, with the spirals segmented into short pieces. The other (fig. 35), with slender but continuous spirals, has been given no definite position either. There are four longer spirals and three shorter ones. If it is not one of the second telophase figures, it must be classified with the regression chromosomes, and should be followed closely by the spirals shown in fig. 36. 1931I TUAN— GASTERIA CHROMOSOMES 57 POLYSPORY In several preparations, 8-celled spore groups are found among other abnormal groups containing from five to many cells in place of the niormal quartet. It has been suggested that the lagging of chromosomes at first metaphase and at many other stages may pro- duce extra nuclei, therefore all the supernumerary cells in any of these groups may be considered to be the natural consequence of such abnormalities. A detailed consideration of the 8-celled groups strengthens the assumption that third division does occur in some of the cells. Eight nuclei of equal size frequently occur in 8-celled groups (fig. 12), while one or two small extra nuclei (fig. 14) are also frequently present. It is easy to account for the small nuclei as the result of misplaced chromosomes, but as to how the eight equal sized nuclei developed, there is only one reasonable explanation; that is, that they are the product of a third division. The evidence in support of the presence of a simple third division may be summed up in a few words. As soon as the chromosomes are formed in each of the four nuclei of a quartet, the nuclear membrane disappears, although the nucleolus, formed at the end of the telophase, is still obvious (fig. 7). The chromosomes, usually seven in number, spread apart in the cell without sign of spindle formation. The next stage, thus far observed, involves cleavage of various depths along the cell wall and cytoplasm. Whether this is the mode of the third division or not is without further proof, and the method by which these chromosomes are distributed to the daughter nuclei is another un- answered question. Additional work on the pollen mother cells of this plant may bring to light more evidence. Role of nucleolus From the resting cells up to the diakinesis stage, the nucleolus remains in the nucleus with little change. Its diameter is estimated as 2.5-3.5M (figs. IS, 16). It disappears almost simultaneously with the nuclear membrane as the chromosomes are arranged for the first division metaphase. It is observed again as a faintly stamed body at the end of the first telophase, with a much reduced diameter (i-i.5m). It increases in size within the nucleus containmg the first type of constricted chromosomes. At this stage it measures S8 BOTANICAL GAZETTE [SEPTEMBER IO31I TUAN— GASTERIA CHROMOSOMES 59 N about 2 M in diameter (fig. 9). With the other type of division, where the interphase and prophase are omitted, the nucleolus does not appear until the second telophase is far under way. It shows the same diameter as before; that is, it first appears as a body about I /x across, and then increases in bulk when the third division chro- mosomes are formed (fig. 7). Its definite function in this process cannot be accounted for; however, the interpretation that the nu- cleolus has some connection with the chromosomal appendages is eUminated. In this case it occurs simultaneously with the fully de- veloped chromosomes, therefore it is not converted into any of these appendages. Chromosome structure Since the structure of chromosomes has already been discussed, only a few outstanding points will be added here. On the metaphase plate the chromosomes are heavily stained. From haematoxylin preparations, the spiral structure is not very distinct, but the num- ber of coils was easily ascertained by the nodule-like structures which bulged out at places along the elastic matrix. It is estimated that in the longer chromosomes the spiral has from seven to nine turns, while in the small ones there are only two or three present. In many metaphase cells indication of a longitudinal spHt was not observed. The single nature is retained throughout until the forma- tion of chromosomes with satellite-like appendages. As previously mentioned, this type of chromosome, in its outward appearance, resembles the normal homeotypic chromosome, but from its in- ternal structure the resemblance is entirely lacking. The cause for such difference is probably due to the fact that the prophase, in which the split is supposed to take place, was skipped, and therefore the split was perforce omitted. The significance of this process is not understood, but it confirms the interpretation of Taylor (26) that the double spiral in the homeotypic chromosomes is the con- sequence of a longitudinal split completed at the second prophase. The internal structure of those chromosomes with giant heads is rather obscure. In many well differentiated cells, it is seen as a comparatively lighter central core with a densely stained peripheral region (fig. 18), containing well spaced nodules in two rows, a struc- ture which makes the chromosomes appear double. When the chro- mosomes come to the second anaphase (fig. 26) the same kind of ap- parent doubleness is maintained, in addition to the vacuoles and anastomoses. Discussion and historical review Several diverse problems are involved in the abnormal behavior of Gasteria chromosomes: (i) the occurrence of three successive di- visions in microsporogenesis; (2) the appearance of the heterotypic type of chromosomes in the second division; (3) the exaggerated constrictions of the chromosomes; (4) the production of polyspores after the close of meiotic activity; (5) the regression of chromosomes from different stages of chromosomal development to a false resting stage; and (6) the structure of chromosomes. Literature recording three or more divisions at the time of meiosis is scattered. Both in spore formation in Ascomycetes and oogenesis in Fucaceae, as well as in embryo sacs, it is well known that mitoses may occur after meiosis before the full number of cells is completed. The occurrence of the division at very different periods in the mor- phological life cycle suggests that the divisions resulting in polyspory are not unforeshadowed and altogether new in higher plants. Re- cently Stow (18), by applying a higher temperature to growing bulbs of Hyacinthus, obtained multinucleate giant pollen grains. He found that cells dividing three times in succession produce struc- tures resembling embryo sacs, while those dividing four or five times give the same kind of irregularity but of a more complicated form. He concluded that the male potency in these cells is reduced by the physical means employed, and thus the female tendency is given a temporary dominance. Although this conclusion is hardly appli- cable to this case of Gasteria, where signs of embryo sac formation are entirely lacking, it suggests the possibility that material for this study was injured by some similar agents in the greenhouse before fixation. As to the appearance of a similar type of chromosomes in the first and second division during meiosis, no corresponding case can be found among plants. Among certain forms of animals such a condi- tion is of common occurrence, therefore it is rather hasty to estabhsh any co-relation here. Constrictions and satellites have been reported frequently as a 6o BOTANICAL GAZETTE [SEPTEMBER 1931] TUAN— GASTERIA CHROMOSOMES 61 permanent morphological character for individual chromosomes. Taylor (21, 22, 23, 24, 25) identified certain chromosomes in many forms by the constrictions and satellites. Sveschnikova (19) classi- fied the genus upon the size of chromosome heads and number of satel- lites seen in many species of Vicia, Sakamura (15) and many others, by means of the use of chloral hydrate and by following the constric- tions, have settled the disputed counts in several plants and cor- rected the old conception of chromosomal tetrads and transverse divisions. However, the origin of such morphological characters is only partially discussed in these articles. Nawaschin (12) prob- ably contributed the first interpretation that nucleolar material is responsible for the satellites. Taylor (22) and Darlington (3) claimed that the satellite is the natural result of an extreme type of subterminal constriction. In the present case, it is seen that the sateUite-like appendages are the direct production of isolated loops of chromatic spirals. Their coexistence with a nucleolus in many of the cells makes it obvious that the nucleolar material of Gasteria plays no direct part in the formation of such appendages. There is an enormous amount of publication on polyspory. Al- most without exception each article deals with hybrids or polyploid species. Taking the rose family for example, Erlanson (4), in a study of American wild roses, found that polyspory is a common phenomenon among spontaneous hybrids or descendants of hybrids ; Shoemaker (17) arrived at the same conclusion with species of cul- tivated apples; Longley (9, 10) discovered the stability of diploid species in Crataegus and Ruhus, while "The polyploid species are characterized by striking irregularities in chromosome distribution and irregular pollen-formation, leading frequently to polycary and polyspory." Investigations on Poteniilla (14) and many related gen- era indicated the same results. In Gasteria, Taylor (21) has indi- cated the hybrid origin of much of the material in cultivation, and questioned the authenticity of specific names as commonly used. There remains Uttle doubt that the species studied is by no means pure, and perhaps some of the abnormahties can be explained on account of genetic weaknesses. In connection with the regression of chromosomes, experiments by Chambers (2) are especially interesting. He demonstrated by microdissection that the reversion of jelly-like chromosomes to a more or less homogeneous sol could be brought about by mechanical injury. With fixed material, degeneration of quartet nuclei is de- scribed in several cases. In Melilotus alba, Rogers (13) and Cas- tetter (i) reported disintegration of three nuclei in quartets and the formation of giant pollen grains by the fourth nucleus. In Carex the same condition of disintegration of certain quartet nuclei and giant pollen formation is given by Juel (5). From these observa- tions it may be inferred that disintegration of nuclei is a normal phenomenon in certain species, and in some others it may be the result of mechanical injury. Kaufmann (6, 7), KuwADA (8), Sharp (16), and Taylor (26) have already offered lengthy reviews on the structure of large chro- mosomes. The discovery of spiral chromonemata and interpretation controverting the older investigators have also been recorded many times. It now seems that the structure of the larger chromosomes is understood to a degree. What remains to be investigated is the structure of smaller chromosomes. With the aid of new and more accurate technique, it is certain that many of the observations of larger chromosomes will be linked together into a coherent interpre- tation when the smaller chromosomes begin to attract the attention of the more skilled workers. So far as behavior of the chromosomes is concerned, the pecuUar cases of Gasteria recorded here are exceedingly interesting as well as perplexing. In their natural environment, such unusual behavior is perhaps being reported for the first time. However, judging from the abnormahties induced by chloral hydrate (15), excessive heat (18), and mechanical injuries (2), it is possible that some physiologi- cal factors are involved. On the other hand, it is probable that the material used is from a certain hybrid species of Gasteria, as shown by the polyspory and lagging of chromosomes at different stages of chromosomal development. It would be interesting from both phy- siological and cytological points of view, therefore, to conduct some parallel experiments with well controlled factors, whereupon it would not be difficult to learn some of the causes of the disturbance. 62 BOTANICAL GAZETTE [SEPTEMBER 193 1] TUAN— GASTERIA CHROMOSOMES 63 ill Summary 1. Behavior of normal reduction chromosomes of Gasteria as de- scribed by Taylor is cited and summarized. 2. Three types of abnormal meiotic and postmeiotic chromosomes of the same plant are described. One type, represented by closely coiled second metaphase chromosomes, is directly derived from the heterotypic chromosomes by an omission of the first interphase and second prophase. The other type, represented by giant headed chro- mosomes, is derived from the first metaphase chromosomes through a partial telophasic disorganization. The third type, represented by heavily constricted chromosomes in binucleated cells, is derived from the first metaphase chromosomes through a short lived inter- phase and breakdown of the nuclear membrane. The constrictions are much exaggerated and resemble the satellites in the somatic chromosomes. 3. The spiral which is involved in the structure of these types is single through the omission of the first interphase and second pro- phase. 4. Formation of third division chromosomes and 8-celled groups is described. This condition is thus far unique in higher plants. 5. Abnormal diakinetic chromosomes and chromosomes at other stages are able to assume a false resting stage after a brief process of disintegration of the chromatic material. 6. A nucleolus was seen in nuclei with fully developed chromo- somes; it was not converted into any of the chromosomal append- ages. It is a pleasure to acknowledge the guidance of Professor Wm. Randolph Taylor, under whose supervision this study was con- ducted. The writer also wishes to express his hearty thanks to Pro- fessor Conway Zirkle and Dr. John M. Fogg Jr. for their criti- cisms and suggestions regarding the manuscript. TsiNG HuA University Peiping, China [Accepted for publication December j8, 1Q30] LITERATURE CITED 1. Castetter, E. F., Studies on the comparative cytology of the annual and biennial varieties of Melilotus alba. Amer. Jour. Bot. 12:270-286. 1925. 2. Chambers, R., Some physical properties of the cell nucleus. Science 40: 824-827. 1914. 3. Darlington, CD., Chromosome studies in the Scilleae. Jour. Genetics 16:237-251. 1925. 4. Erlanson, Eileen W., Cytological conditions and evidences for hybridity in North American wild roses. Box. Gaz. 87:443-509. 1929. 5. JuEL,H.O.,BeitragezurKenntnissderTetradenbildung. Jahrb.Wiss. Bot. 35:626-659. 1900. 6. Kaufmann, B. p., Chromosome structure and its relation to the chromo- some cycle. I. Somatic mitoses in Tradescantia pilosa. Amer. Jour. Bot. 13:59-80. 1926. tj^ ^ Chromosome structure and its relation to the chromosome cycle. II. Podophyllum peltatum. Amer. Jour. Bot. 13:355-363- 1926. 8. KUWADA, Y., On the spiral structure of chromosomes. Bot. Mag. Tokyo 41:100-109. 1927. 9. LoNGLEY, A. E., Cytological studies in the genus Rubus. Amer. Jour. Bot. 11:249-282. 1924. 10. , Cytological studies in the genus Crataegus. Amer. Jour. Bot. 11: 295-316. 1924. 11. McClung, C. E., Handbook of microscopical technique. Paul B. Hoeber Inc. New York. 1929. 12. Nawaschin, M., Morphologisches Kernstudium der Crepis-Arten in Bezug auf die Artbildung. Zeitschr. Zellforsch. Mikro. Anat. 2:98-111. 1925. 13. Rogers, W. E., Notes on Melilotus alba. Proc. la. Acad. Sci. 24:415-482. 1917- 14. Roscoe, Muriel V., Meiotic irregularities in a gigas form of Potentilla anserina. Box. Gaz. 84:307-316. 1927. 15. Sakamura, T., Experimentelle Studien iiber die Zell- und Kernteilung mit besonderer Rucksicht auf Form Grosse und Zahl der Chromosomen. Jour. Coll. Sci. Imp. Univ. Tokyo 39:1-221. 1920. 16. Sharp, L. W., Structure of large somatic chromosomes. Box. Gaz. 88:349- 382. 1929. 17. Shoemaker, J. B., Pollen development in the apple, with special reference to chromosome behavior. Box. Gaz. 81:148-172. 1926. 18. Sxow, ISAMU, Experimental studies on the formation of the embryo sac- like giant pollen grain in the anther of Hyacinthus orientalis. Cytologia 1:417-439. 1930. 19. SvESCHNiKOVA, I. N., Karyological studies in Vicia. Bull. Appl. Bot. Genet. Pt. Breeding 17:37-72. 1927- • BOTANICAL GAZETTE, XCII PLATE I 64 BOTANICAL GAZETTE [SEPTEMBER 20. Taylor, W. R., The smear method for plant cytology. Box. Gaz. 78: 236- 238. 1924. 21. , Cytological studies on Gasteria. I. Chromosome shape and indi- viduality. Amer. Jour. Bot. ii:5ih3i. 1924. 22. , The chromosome morphology of Velthemia, Allium, and Cyrtanthus. Amer. Jour. Bot. 12:104-116. 1925. 23. , Cytological studies on Gasteria. II. A comparison of chromosomes of Gasteria, Aloe, and Hawarthia. Amer. Jour. Bot. 12:219-223. 1925. 24. , Chromosome constrictions as distinguishing characteristics in plants. Amer. Jour. Bot. 12:238-244. 1925- 25. , Chromosome morphology in Frilillaria, Alstroemeria, Silphium, and other genera. Amer. Jour. Bot. 13:179-193- 1926. 5. ^ Chromosome studies on Gasteria. III. Chromosome structure dur- ing microsporogenesis and the postmeiotic mitosis. Amer. Jour. Bot. (In Press). 1 93 1. 27. TuAN, H. C, A new method for safranin differentiation. Stain Tech. 5 : 103- 107. 1930. 28. , Picric acid as a destaining agent for iron alum haematoxylin. Stain Tech. 5:135-138. 1930. EXPLANATION OF PLATES I, II All drawings were made with the aid of Zeiss 1.5 mm. apochromatic objec- tive and X 20 compensation ocular, giving an approximate magnification of 4500. PLATE I Fig. 15. — Open spireme nucleus. Fig. 16. — Nucleus at diakinesis stage. Pig jy —Segment of diakinetic chromosomes showing first sign of regression. Fig. 18.— Giant headed chromosomes showing structural detail. Fig. 19.— Mid-telophase chromosomes showing loosely connected spirals. Fig. 20.— First telophase chromosomes (a, b, c) showing origin of con- strictions. Fig. 21.— Early telophase nucleus showing loosening of spirals and forma- tion of nuclear membrane at side near polar region. Fig. 22.— Third division chromosomes (a, b, c, d) showing process of re- gression. Fig. 23.— Late telophase spiral showing anastomoses and vacuolation. Fig. 24. — Two sets of spiral chromosomes just before second metaphase. Fig. 25. — Second anaphase of spiral chromosomes. Fig. 26. — Second anaphase of giant headed chromosomes. PLATE II Fig. 27. — First metaphase chromosomes at equatorial plate stage. Fig. 28. — Spirals in quartet nucleus, seven can be counted. 18 17 21 19 TUAN on GASTERIA BOTANICAL GAZETTE, XCII PLATE I 64 BOTANICAL GAZETTE [SEPTEMBER 20. Taylor, W. R., The smear method for plant cytology. Box. Gaz. 78:236- 238. 1924. 21. , Cytological studies on Gasteria. I. Chromosome shape and indi- viduality. Amer. Jour. Bot. 11:51-61. 1Q24. 22. , The chromosome morphology of Velthemia, Allium, and Cyrtanthus. Amer. Jour. Bot. 12:104-116. 1925. 23. , Cytological studies on Gasteria. II. A comparison of chromosomes of Gasteria, Aloe, and Hawarthia. Amer. Jour. Bot. 12:219-223. 1925. 24. , Chromosome constrictions as distinguishing characteristics in plants. Amer. Jour. Bot. 12:238-244. 1925- 25. , Chromosome morphology in Frilillaria, Alstroemeria, Silphium, and other genera. Amer. Jour. Bot. 13:179-193- 1926. 6, ^ Chromosome studies on Gasteria. III. Chromosome structure dur- ing microsporogenesis and the postmeiotic mitosis. Amer. Jour. Bot. (In Press). 1931- 27. TuAN, H. C, A new method for safranin differentiation. Stain Tech. 5 : 103- 107. 1930. . , 1 i- 28. , Picric acid as a destaining agent for iron alum haematoxylm. Stain Tech. 5:135-138. 1930- EXPLANATION OF PLATES I, II All drawings were made with the aid of Zeiss 1.5 mm. apochromatic objec- tive and X 20 compensation ocular, giving an approximate magnification of 4 SCO. PLATE I Fig. 15. — Open spireme nucleus. Fig. 16. — Nucleus at diakinesis stage. Fig. 17.— Segment of diakinetic chromosomes showing first sign of regression. Fig. 18.— Giant headed chromosomes showing structural detail. Fig. iq.— Mid-telophase chromosomes showing loosely connected spirals. Fig. 20.~First telophase chromosomes (a, h, c) showing origin of con- strictions. Fig. 21.— Early telophase nucleus showing loosening of spirals and forma- tion of nuclear membrane at side near polar region. Fig. 22.— Third division chromosomes {a, b, c, d) showing process of re- gression. Fig. 23.— Late telophase spiral showing anastomoses and vacuolation. Fig. 24.— Two sets of spiral chromosomes just before second metaphase. Fig. 25. — Second anaphase of spiral chromosomes. Fig. 26. — Second anaphase of giant headed chromosomes. PLATE II Fig. 27. — First metaphase chromosomes at equatorial plate stage. Fig. 28.— Spirals in quartet nucleus, seven can be counted. i 16 18 B 20 19 B C 17 21 24 TUAN on GASTERIA INTENTIONAL SECOND EXPOSURE BOTANICAL GAZETTE, XCII a- PLATE II \ 29 32 B D E \ N V e». 34 ''^i? B 38 X \' 39 // \ 40 TUAN on GASTERIA BOTANICAL GAZETTE, XCII PLATE II '- fTlflftlPiirNftl ^•^ \ 29 •- •' .^ '. f- ^ ^ -^ 31 32 B C D E r ^. t .IJ' 35 B ^i J -J-. ^ X •*.. ^;^.- ^ \ \ A %. ^ • 39 \ ,' ? 40 Ffrfi TUAN on GASTERIA INTENTIONAL SECOND EXPOSURE I« i 1931I TUAN— GASTERIA CHROMOSOMES 65 Fig. 29. — Chromatic thread from regression diakinetic chromosome. Fig. 30. — Two sets of second metaphase dyads derived directly from first metaphase tetrads showing spiral structure in each of chromatids. Fig. 31. — ^Late anaphase daughter group showing separation of dyad part- ners. Fig. 32. — Tetrad threads from regression diakinetic chromosomes. Fig. 33. — Selected second telophase giant headed chromosomes (a, b, c, d, e) showing disorganization of chromatin material; head still recognizable in a. Fig. 34. — Spirals from quartet nucleus. Fig. 35. — Seven continuous spirals from another quartet nucleus. Fig. 36. — Quartet nucleus showing regression of third division chromosomes. Fig. 37. — First telophase spiral showing anastomoses. Fig. 38. — Same as fig. 37. Fig. 39. — Quartet nucleus at early telophase, seven spirals can be counted. Fig. 40. — Same as fig. 36; b shown at resting stage. Reprinted from Bartonia, No. 13, 1931. PHILADELPHIA BOTANICAL CLUB 19 The Eastern Long-styled Phloxes, part i' Edgar T. Wherry Continuing the discussion of the eastern species of Phlox, the section now to be taken up differs from the one next pre- ceding in that the sepals are usually united to half their length or more, and the stamens and styles are nearly or quite as long as the corolla-tube. Peter^ termed this section the Beptantes, but as the species name reptans has been super- seded, another section name seems preferable, and Ovatae is here proposed. In the order believed to represent evolution- ary progress in the group — that of gradual loss of prostrate stems, decrease in persistence of leaves and increase in com- plexity of inflorescence, — ^the species are : PHLOX, SECTION OVATAE: KEY TO SPECIES Prostrate stems well developed, rooting and bearing flowering shoots at nodes; lower leaves spatulate, evergreen 8. P. stolonifera Prostrate stems poorly developed or lacking, exceptionally rooting at nodes; lower leaves never typically spatulate. Flowering shoots mostly arising from clusters of leaves terminating decumbent stems; nodes few; calyx averaging 10 mm. long. Lower leaves ensiform, evergreen; upper lanceolate; inflores- cence conspicuously glandular ^ 9. P. huclcleyi Lower leaves elliptic, marcescent; upper elliptic to ovate; in- florescence minutely pubescent 10. P. ovata Flowering shoots mostly arising from leafless rootstocks; nodes numerous; calyx averaging less than 10 mm. long. Rootstock short, sending up green or diffusely red-mottled stems; cymes grouped in a corymb or broad corymbose panicle with more or less elongate branches. Upper leaves lanceolate to ovate; calyx 6-11 mm. long; range centering in the mountains, locally extending into lowland provinces 11. P. Carolina Upper leaves linear to lanceolate; calyx 6-8 mm. long; range centering in the lowlands, locally extending to moderate elevations 12. P. glaherrima Bootstock elongated, curving upward into a strongly red- mottled or exceptionally green stem; cymes in a narrow con- ical or cylindrical panicle 13. P. maculata 1 Contribution from the Botanical Laboratory of the University of Pennsylvania. Previous articles in the series have appeared in Bartonia 11: 5. 1929 and 12: 24. 1931. 2 In Engler & PrantPs Pflanzenfamilien 43a: 46. 1891. (18) The first two species listed in the synopsis on the opposite page are readily distinguishable, but the others exhibit more or less intergradation. As the treatment of these adopted here differs in some respects from that followed in current manuals and floras, a brief discussion of the method used in deciding what species should be recognized may well be pre- sented. Each of the commonly accepted species was studied in the field at ten or more localities. Within any one colony of Phlox ovata the individual plants proved to be constant in having a small number of nodes on the stem below the in- florescence and a relatively long calyx. Marked variation was shown, on the other hand, in leaf -shape, many plants having much narrower leaves than the species name would imply. All specimens with numerous nodes and short calyx were accordingly withdrawn from the Phlox ovata covers in the herbaria being consulted. In P. glaherrima constancy was found in the essentially lanceolate leaf outline, and short calyx, so material with more elliptic leaves and longer calyx was similarly withdrawn. Again the species name turned out to be rather inappropriate, for the majority of the plants showed at least a small amount of pubescence. Colonies of P. maculata showed a strikingly constant ten- dency to cylindrical or at most narrowly conical inflorescence, and from its covers were accordingly taken plants exhibiting a broader cone or a corymb of flowers. Here the character to which the species name refers could not be safely used in diagnosis, since several other Phloxes have the stem more or less maculate. When the aberrant specimens from these sources were brought together, they proved to resemble one another in many respects, and individuals could readily be found among them which corresponded closely to the type specimen of Linne's P. Carolina. As the withdrawal of this from the accepted list of species had been due to Gray, who had no adequate field acquaintance with the plants, its reinstatement seemed fully justified. I t 20 PEOCEEDINGS OP THE I 8. Phlox stolonifera Sims. Creeping Phlox. Plate 4. History.— So far as recorded this Phlox was first observed, in Georgia, by the horticultural collector John Eraser in 1786, and living material sent by him to England in 1801 formed the basis of the specific description by Sims.^ It was also found about the same time by Michaux^ and named P. reptansj this has been widely used, but since Sims's name has a year's priority, Michaux's must be relegated to synonymy. Subse- quent names for the same species were P. ohovata Muhlenberg ex Willdenow,^ P. prostrata Alton,* and P. crassifolia Lod- diges.^ The last, as well as the combination P. stolonifera crassifolia Don,^ represented the purple color-form, which is the more frequent and wide-spread ; the original collection by Fraser, however, was of a form with violet corolla which occurs locally in the Blue Eidge, and the name P. stolonifera forma violacea has recently been applied to this by Peattie.^ As pointed out under P. amoena in the preceding article in this series,^ a showy-flowered Phlox which has long been in cultivation combines the characters of P. sululata sjid P. stolonifera in such a striking way as to clearly indicate its origin as a hybrid between these species. This has been named successively P. procumhens Lehmann,^ P. suhulata 3 latifolia Bentham,^^ *'P. verna'' Hort. ex Vilmorin" and '*P. amoena" Hort., not Sims ; and these names, either alone or in various combinations, are sometimes mistakenly applied to P. stolon- ifera itself. As an example of the latter procedure, there may be cited the naming of what appears to be a color-form of the species **P. procumbens coerulea'' Hort. ex Crockett.^^ 1 Curtis 's Botanical Magazine 16: pi. 563. 1802. 2 Flora Boreali- Americana 1 : 145. 1803. 3 Enum. PI. Hort. Reg. Berol. 201. 1809. 4 Hortus Kewensis, ed. 2. 1: 326. 1810. 5 Botanical Cabinet 16 : pi. 1596. 1829. 6 In Sweet's British Flower Garden, ser. 2. 3: pi. 293. 1835. 7 Journ. EUsha Mitchell Sci. Soc. 45 : 266. 1930. 8 Bartonia 12 : 53. 1931. » Index Sem. Hamburg 1828: 17; Linnaea 5: 383. 1830. 10 In DeCandoUe's Prodromus 9: 306. 1845. 11 Fleurs Pleine Terre, ed. 2. 682. 1866. 12 Amer. Botanist 30: 159. 1924. . , Bartonia, No. 13 m Plate 4 #.^ ' ^n ;n ^rmv ' u^ I ■>*%. ■f ~i* • — m-': .'"-•* « ♦♦ ^i" Fig. 1. Phlox stolonifera. East of Mountain Lake, Garrett County, Maryland. Fig. 2. Phlox stolonifera. In cultivation; originally from North Carolina. 20 PROCEEDINGS OF THE ]>ART0N1A, No. 13 Plate 4 m m 8. Phlox stolonifera Sims. Creeping Phlox. Plate 4. History.— ^0 far as recorded this Phlox was first observed, in Georgia, by the horticultural collector John Eraser in 1786, and living material sent by him to England in 1801 formed the basis of the specific description by Sims.^ It was also found about the same time by Michaux^ and named P. reptans; this has been widely used, but since Sims's name has a yearns priority, Michaux's must be relegated to synonymy. Subse- quent names for the same species were P. ohovata Muhlenberg ex Willdenow,^ P. prostrata Alton,* and P. crassifoUa Lod- diges.^ The last, as well as the combination P. stolonifera crassifoUa Don,« represented the purple color-form, which is the more frequent and wide-spread ; the original collection by Eraser, however, was of a form with violet corolla which occurs locally in the Blue Ridge, and the name P. stolonifera forma violacea has recently been applied to this by Peattie.'^ As pointed out under P. amoena in the preceding article in this series,^ a showy-flowered Phlox which has long been in cultivation combines the characters of P. suhulata and P. stolonifera in such a striking way as to clearly indicate its origin as a hybrid between these species. This has been named successively P. procumhens Lehmann,® P. suhulata 3 latifolia Bentham,^^ *'P. verna'^ Hort. ex Vilmorin^^ and ''P. amoena" Hort., not Sims ; and these names, either alone or in various combinations, are sometimes mistakenly applied to P. stolon- ifera itself. As an example of the latter procedure, there may be cited the naming of what appears to be a color-form of the species **P. procumbens coerulea'' Hort. ex Crockett.^^ 1 Curtis 's Botanical Magazine 16: pi. 563. 1802. 2 Flora Boreali- Americana 1 : 145. 1803. 3 Enum. PI. Hort. Reg. Berol. 201. 1809. 4Hortus Kewensis, ed. 2. 1: 326. 1810. 5 Botanical Cabinet 16 : pi. 1596. 1829. Gin Sweet's British Flower Garden, ser. 2. 7 Journ. Elisha MitcheU Sci. Soc. 45 : 266. 8 Bartonia 12 : 53. 1931. 9 Index Sem. Hamburg 1828: 17; Linnaea 5: 383. 1830. 10 In DeCandolle's Prodromus 9: 306. 1845. 11 Fleurs Pleine Terre, ed. 2. 682. 1866. 12 Amer. Botanist 30 : 159. 1924. 3: pi. 293. 1835. 1930. H, .'^ aw '^^^^''^'^ . ■:i.^i' . 4^ *^ <^«^ !■ ■ aV-^* ' . 4..: .•• ♦ ,N4 ^ ! Fig. 1. PIih).r stolonifera. East of Mountain Lake, Garrett Countv, Maryland. Fig. 2. Phlox sfolouifrra. In cultivation; originally from North Carolina. m INTENTIONAL SECOND EXPOSURE ■ PHILADELPHIA BOTANICAL CLUB 21 Geography. — Phlox stolonifera ranges from Georgia to Ohio and Pennsylvania, chiefly at high altitudes, though not on mountain summits. It no doubt survived glaciation in some portion of this area, but in its subsequent migration has not quite reached the Wisconsin terminal moraine, which is shown on the accompanying map by a dotted line. The moun- tain front, marked by a row of A 's, has been a barrier to its eastern spread (except in the south), but westward it has de- scended from the mountains to the ravines of southern Ohio. Fig. 1. Distribution of Phlox stolonifera, [Alabama: Reported by Mohr^ from Cullman County, but the specimens so labelled, preserved in the U. S. National Herbarium, represent P. Carolina.] [Arkansas : NuttalP and Rafinesque, according to Brand,^ listed this species as present in Arkansas, but in the lack of specimens collected by them or by subsequent botanists, a mis- identification is believed to have occurred.] [Florida: In the Short herbarium, now at the Academy of Natural Sciences of Philadelphia, there is a specimen which undeniably represents this species labelled: ** Phlox — ^near reptans. Cliffs of Aspalaga. (an P. divaricata ? S.) Florida. Dr. A. W. Chapman.'' As, however. Chapman* did not in- 1 Plant Life of Ala. 686. 1901 (as P. reptans), 2 Trans. Amer. Phil. Soc. 5: 196. 1837. 3 In Engler 's Pflanzenreich IV. 250 : 63. 1907. * Flora Southern States 338. 1860. 22 PROCEEDINGS OF THE PHILADELPHIA BOTANICAL CLUB 23 m III T? % elude Florida within the range of the species, and it has not been collected there by any one else, the specimen may really have come from the cliffs of some mountain stream, and the locality names have got confused in making out the label to send to Short.] Georgia : As noted under History, this plant was first dis- covered by Fraser in Georgia, although the exact locality is not recorded. It must be rare there, as herbaria contain speci- mens from but two counties, Fannin and Wilkes. [Illinois: Brand^ included this state in the range of P. stolonifera, and even added an exclamation point (!) to indi- cate certainty, but in the absence of specimens the report is not here credited.] Kentucky: Limited to the eastern end, specimens being preserved from Licking Kiver, Morgan County. Maryland : Not listed in the report on the plant life of the state, but frequent in Garrett' County. [Mississippi : Included by Lowe,^ but specimens so labelled, seen in the State University Herbarium, proved to be P. di- varicata with the stolons particularly well developed.] [Missouri: Another unsubstantiated report by Brand.] North Carolina : Fairly common in mountain woods, there being 7 county records : Avery,' Buncombe, Madison, Mecklen- burg, Polk,' Swain/ and Watauga. Ohio : Occasional in the cool ravines of the southern part of the state, notably in Hocking and Jackson' counties. Pennsylvania: Locally abundant in the Appalachians, in the counties: Armstrong, Blair, Cambria, Center' (northeast- ern limit for the species), Clearfield, Fayette, Huntingdon, Somerset, and Westmoreland. South Carolina : Rare, and reported only from Greenville County, by Peattie.^ Tennessee: Known to have been collected in 3 counties toward the eastern end : Cocke, Polk, and Sevier. [Vermont: Escaped in Lamoille and Rutland counties.] iln Engler's Pflanzenreich IV. 250: 63. 1907. 2 Plants of Mississippi 233. 1921 (as P. reptans). 3 Journ. Elisha Mitchell Sci. Soc. 45 : 266. 1930. Virginia : Present in five Appalachian counties, Bath,' Car- roll, Grayson, Smyth, and Wythe. West Virginia : Apparently reaches maximum development here, the county list being: Cabell, Fayette, Gilmer, Grant,' Greenbrier,' Mason, Mercer, Monongalia, Monroe, Pendleton/ Pocahontas,' Preston, Taylor, Tucker, and Upshur. Ecology. — Phlox stolonifera is typically an occupant of rich woods in non-calcareous regions, where its stems creep through the subacid leaf -litter. It also occasionally pushes out onto alluvial flats along streams, or rarely occupies rock ledges. On the whole, however, it is to be classed as a mesophytic forest herb. Its flowering season is vernal, beginning in late April, before the leaves of the deciduous trees are fully de- veloped, and extending through May into early June. The long style brings the stigmas into the midst of the anthers, but protandry prevents self-pollination, and the long-tubed bright purple flowers attract butterflies which transfer pollen from one clump to another. The development of ability to propagate vegetatively has apparently been accompanied by lessened tendency to reproduce through seeds, and few of its ovules seem to mature. Variation. — The variations shown by this species are less marked than in most Phloxes, and it does not grade toward other members of the section. Individual clumps differ in vigor, and correspondingly in size of leaves and floral parts, but the range is only moderate. As already stated under History, there are two distinct types of corolla-color, phlox- purple (Ridgway 65 b), which is the more wide-spread, and light violet (59 b), which appears locally in southern North Carolina and adjacent portions of other states. Few pallid phases of either of these have been noted, and albinos are practically unknown. As in previous papers of this series, color-forms will not be given technical names. Cultivation. — The early references to this species noted under History concerned cultivated material, but most gar- dens are not sufiiciently shady or rich in acid humus to suit it, and it is rarely grown. The beauty of its flowers should lead, however, to its more extensive use. '»• 24 PROCEEDINGS OF THE Bartonia, No. 13 Plate 5 \ Fig. 2. Distribution of Phlox hucMeyi, 9. Phlox buckleyi Wherry. Sword-leap Phlox. Plate 5. History. — This highly distinct species was collected by S. B. Buckley at White Sulphur Springs, West Virginia, in June, 1838, but lay unnoticed in his herbarium for 80 years. It was rediscovered at the same place by Miss Marian S. Franklin of Lewisburg about 1919, and several new stations were located in 1929, encouraging its description and naming.^ Geography, — At the time the species was described six lo- calities were known, equally divided between Virginia and West Virginia. An additional one has since been found, but the plant remains the most restricted endemic of all our eastern Phloxes, as brought out in the map herewith. Virginia: Now known in iL^two counties, Alleghany' and ■^^^■""**^Bath./ West Virginia: Found thus far only in Greenbrier' Co. Ecology. — The chief reason for the geographic restriction of this species is seemingly its requirement for a special type of habitat, for in all seven known localities it grows in a gravelly soil derived from a particularly slabby Devonian shale. It is, however, not a pioneer on shale-slopes, being unable to come in until after a thin woodland cover has de- veloped and the soil has become rich in subacid humus. The denser shade of climax forests is also unfavorable to it, and it dies out as they mature. Its thick narrow persistent leaves lead to its classification as moderately xerophytic. The bloom- ing period extends from mid-May to mid-June, the bright phlox-purple flowers attracting insects as do those of the preceding species. Variation. — Only minor variations, of nutritional origin, have been observed. Cultivation. — The difficulty of matching the native habitat conditions of this plant is likely to prevent its ever being much cultivated. 1 Journ. Wash. Acad. Sci. 20 : 25. 1930. Fig. 1. Phlox hncTcleyi. Southeast of Alleghany Station, Alleghany County, Virginia. Fig. 2. Phlox huclleyi. Southeast of Caldwell, Greenbrier County, West Virginia. 24 PROCEEDINGS OF THE Bartonia, No. 13 Plate 5 m 9. Phlox buckleyi Wherry. Sword-leaf Phlox. Plate 5. History. — This highly distinct species was collected by S. B. Buckley at White Sulphur Springs, "West Virginia, in June, 1838, but lay unnoticed in his herbarium for 80 years. It was rediscovered at the same place by Miss Marian S. Franklin of Lewisburg about 1919, and several new stations were located in 1929, encouraging its description and naming.^ Geography. — At the time the species was described six lo- calities were known, equally divided between Virginia and West Virginia. An additional one has since been found, but the plant remains the most restricted endemic of all our eastern Phloxes, as brought out in the map herewith. Virginia: Now known in 4'-'-'-^-^W two counties, Alleghany^ and Fig. 2. Distribution of Phlox hucMeyu Bath.^ West Virginia: Found thus far only in Greenbrier' Co. Ecology. — The chief reason for the geographic restriction of this species is seemingly its requirement for a special type of habitat, for in all seven known localities it growls in a gravelly soil derived from a particularly slabby Devonian shale. It is, however, not a pioneer on shale-slopes, being unable to come in until after a thin woodland cover has de- veloped and the soil has become rich in subacid humus. The denser shade of climax forests is also unfavorable to it, and it dies out as they mature. Its thick narrow persistent leaves lead to its classification as moderately xerophytic. The bloom- ing period extends from mid-May to mid-June, the bright phlox-purple flowers attracting insects as do those of the preceding species. Variation. — Only minor variations, of nutritional origin, have been observed. Cultivation. — The difficulty of matching the native habitat conditions of this plant is likely to prevent its ever being much cultivated. 1 Journ. Wash. Acad. Sci. 20 : 25. 1930. Fig. 1. Phloa' hiicl-leyi. Southeast of Alleghany Station, Alleghany County, Virginia. Fig. 2. Phlox hucklcyi. Southeast of Caldwell, Greenbrier County, West Virginia. INTENTIONAL SECOND EXPOSURE I>AKT()NIA, No. 13 Plate 6 PHILADELPHIA BOTANICAL CLUB 25 'I m Fig. 1. Phlox ovnia. West of White Sulpliur Springs, Greenl)rier ('oiinty, West Virginia. Fig. 2. Pltlox ovafa. In cultivation; originally from Virginia. 10. Phlox ovata Linne. Mountain Phlox. Plate G. History. — In Plukenet's^ Mantissa of 1700 there was in- eluded a plant characterized as ''Lychnidea fistulosa Mari- landica, Clinopodii vulgaris folio, flore amplo singulari.'' The figure shows this as a Phlox with remote, ovate, petioled leaves and a single flower subtended by a large bract. Linne^ named it Phlox ovata, basing his diagnosis on the Plukenet figure, although Jackson^ records that the specimen in his herbarium (which has several flow^ers in a corymb) was already there in 1753. Sir Josepli Banks, according to Sims,* recognized that Plukenet 's figure and Linnets diagnosis referred to an ab- normal individual, and extended the name P. ovata to cover plants with corymbose flowers. Michaux,^ on the other hand, proposed P. latifolia for the many-flowered plants he collected in North Carolina. Pursh^ listed both ovata and latifoliay not realizing that they are identical. In the second edition of his Species Plantarum, Linne^ described another Phlox with broad leaves as P. Carolina. Bentham^ considered this to be conspecific with P. ovata, but separated three varieties, applying to one of them the com- bination P. Carolina a ovata. In 1870 Gray® pointed out objections to this procedure, proposing instead P. ovata elatior; and later^^ discarded the name Carolina entirely, as a synonym of P. ovata. In this he has been followed by most botanists, but a study of photographs^^ of the Linnean speci- mens of the two shows that they actually represent distinct species, and P. Carolina will be discussed at length on a sub- sequent page. 1 Mantissa 122, pi. 348, tig. 122-pl. 4. 1700. 2 Species Plantarum (1): 152. 1753. sSuppl. Proc. Linn. Sec. 124: 116. 1912. * Curtis 's Botanical Magazine 15: pi. 528. 1801. 5 Flora Boreali-Americana 1 : 143. 1803. 6 Flora Americae Septentrionalis 1 : 148 & 150. 1814. 7 Species Plantarum, ed. 2. 1 : 216. 1762. 8 In De Candolle's Prodromus 9: 304. 1845. 9 Proc. Amer. Acad. Arts Sci. 8: 249. 1870. 10 Synoptical Flora N. A. 2, pt. 1 : 130. 1878. 11 Kindly obtained for me by Dr. Wm. E. Maxon. Baktonia, No. 13 Plate 6 PHILADELPHIA BOTANICAL CLUB 25 iim'i Fig. 1. Phlox ovata. West of White Sulphur Springs, Greenbrier County, W^est Virginia. Fig. 2. Phlox ovata. In cultivation; originally from Virginia. 10. Phlox ovata Linne. Mountain Phlox. Plate 6. History. — In Plukenet's^ Mantissa of 1700 there was in- cluded a plant characterized as **Lychnidea fistulosa Mari- landica, Clinopodii vulgaris folio, flore amplo singulari." The figure shows this as a Phlox with remote, ovate, petioled leaves and a single flower subtended by a large bract. Linne^ named it Phlox ovata, basing his diagnosis on the Plukenet figure, although Jackson' records that the specimen in his herbarium (which has several flowers in a corymb) was already there in 1753. Sir Joseph Banks, according to Sims,* recognized that Plukenet 's figure and Linne 's diagnosis referred to an ab- normal individual, and extended the name P. ovata to cover plants with corymbose flowers. Michaux,^ on the other hand, proposed P. latifolia for the many-flowered plants he collected in North Carolina. Pursh^ listed both ovata and latifolia, not realizing that they are identical. In the second edition of his Species Plantarum, Linne^ described another Phlox with broad leaves as P. carolvna. Bentham^ considered this to be conspecific with P. ovata, but separated three varieties, applying to one of them the com- bination P. Carolina a ovata. In 1870 Gray^ pointed out objections to this procedure, proposing instead P. ovata elatior; and later^° discarded the name Carolina entirely, as a synonym of P. ovata. In this he has been followed by most botanists, but a study of photographs^^ of the Linnean speci- mens of the two shows that they actually represent distinct species, and P. Carolina will be discussed at length on a sub- sequent page. 1 Mantissa 122, pi. 348, fig. 122-pl. 4. 1700. 2 Species Plantarum (1): 152. 1753. sSuppl. Proc. Linn. See. 124: 116. 1912. * Curtis 's Botanical Magazine 15 : pi. 528. 1801. 5 Flora Boreali- Americana 1 : 143. 1803. 6 Flora Americae Septentrionalis 1 : 148 & 150. 1814. 7 Species Plantarum, ed. 2. 1 : 216. 1762. 8 In De Candolle's Prodromus 9: 304. 1845. eProc. Amer. Acad. Arts Sci. 8: 249. 1870. 10 Synoptical Flora N. A. 2, pt. 1 : 130. 1878. 11 Kindly obtained for me by Dr. Wm. E. Maxon. tl INTENTIONAL SECOND EXPOSURE 26 PROCEEDINGS OF THE PHILADELPHIA BOTANICAL CLUB 27 Geography. — Like Phlox stolonifera, P. ovata is essentially a southern Appalachian plant, but the ranges of the two differ in details, as comparison of their distribution maps will show. The most remarkable feature of the present species consists in its occurrence in an outlying area largely within the glaci- ated territory of Indiana and Ohio. The route by which migration to this area took place is uncertain, but it probably traversed eastern Kentucky. Fig. 3. Distribution of Phlox ovata, [Alabama: The record for Monroe County published by Mohr^ is shown by his material in the National Herbarium to have been based on occurrences of P. Carolina.] [Florida: Brand^ included this state in the range of P. ovata, citing a specimen **Rafinesque n. 12." As there is no other report from anywhere near its boundaries, however, an error in identification or labelling is believed to have oc- curred.] Georgia : Exceptional this far south, specimens having been seen only from Dade County, at the northwest corner of the state. 1 Plant Life of Alabama 685. 1901. 2 In Engler's Pflanzenreich IV. 250: 63. 1907. Indiana: Found locally toward the east side, in Allen, Clark, Jay, Jefferson, and Whitley counties. [Kentucky: Although no specimens have been seen, this species no doubt occurs in the eastern uplands, over which it presumably migrated in reaching the Ohio-Indiana area.] Maryland : Eecorded by Shreve^ from Allegany County. [Massachusetts: Escaped in Berkshire County.] North Carolina: Abundant in the Blue Ridge province, and extending, as do many other mountain plants here, well out into the Piedmont. The county list is: Ashe,^ Avery,' Buncombe,' Burke, Caldwell,' Forsyth,' Franklin,' Jackson, Madison,' Orange,' Polk, Rockingham,' Stokes,' Surry,' Wake,' and Yancey.' Ohio: Specimens have been seen from Fulton, Highland, and Lucas counties, the last being the northernmost point the species is known to reach. Pennsylvania: Occasional in the Appalachians and inner Piedmont, there being six county records: Berks, Center, Franklin, Huntingdon, Mifflin, and York. The first of these represents the eastern limit for the species. [South Carolina: Probably present along the northwest side, although no specimens or dependable records have been seen,] Tennessee: Scattered through the eastern uplands, and known from Cocke, Hamilton, Hawkins,' Knox, and Roane counties. Virginia : Frequent in the Appalachians and southern Blue Ridge, herbaria containing specimens from 15 counties: Alle- ghany,' Bath,' Bedford, Botetourt, Floyd,' Giles, Highland,' Montgomery,' Pittsylvania, Pulaski,' Roanoke,' Rockbridge, Shenandoah, Smyth, and Wythe. West Virginia: Overlooked by Millspaugh^ in listing the plants of the state, but actually not uncommon toward the southeastern side, as the following county list shows: Green- brier,' Hampshire,' Hardy, Mercer,' Monroe,' Morgan, and Pendleton.' 1 Plant Life of Maryland 473. 1910. 2 Living Flora of West Virginia. 1913. 28 PROCEEDINGS OP THE PHILADELPHIA BOTANICAL CLUB 29 •.^ Ecology. — Phlox ovata grows chiefly in open woods or thickets, but occasionally enters alluvial meadows and rarely tracts underlain by calcareous rocks. Its soils may be either high or low in humus, but are almost always subacid in re- action. In successions it comes in fairly early, reaches its best development at an intermediate stage, and dies out as climax forest conditions are approached. It begins to bloom about the first of May, and continues through June or even early July at higher elevations. The relations of its flowers to insects are similar to those of the preceding members of the section. Variation, — Unlike the two species just considered, the present one is markedly variable, especially in foliage char- acters, and unless herbarium specimens are fairly complete it is difficult to distinguish with certainty from the one which follows. The flowering shoots average 40 cm. in height, and are notably f ew^-leaved, there being usually only three or four nodes below the inflorescence proper. Their lowest leaves have long petioles and linear blades, and higher ones exhibit progressive shortening of petioles and widening of blades, until the pair below the bracts may be cordate-clasping and orbicular-ovate. The degree of blade-enlargement varies markedly from one plant to another, however, and is some- times so slight that even the uppermost leaves are still petioled with the blades scarcely wider than linear-elliptic. Stating the situation in another way, measurements of the largest leaf-blades (made on 50 plants) show them to range in length from 50 to 100 mm. and in width from 10 to 30 mm. The first bracts are often smaller than the leaves, but may be larger. On sterile shoots, one or more of which are usually conspicu- ous around the crown of the plant, the leaf -blades show similar variation in width, but petioles are always well-developed. Under such circumstances it is manifestly impossible to draw up any simple description of the leaf -characters of this species, or to depend primarily upon the leaves for constructing a key to the section, as attempted in some manuals. The use of a combination of characters for diagnostic purposes seems un- avoidable. The calyx of Phlox ovata averages longer than that of any other eastern species, and this feature can often be used to recognize it when the specimens lack sterile shoots, although it must be borne in mind that occasional individuals normal in other respects may have an exceptionally short calyx. The usual range in sepal-length is from 8.5 to 12 mm., with 10 mm. most frequent. The sepals are often united to half, but some- times only to 0.4, and again to 0.6 or even 0.8 their length. Although the calyx is normally glabrous, the pedicels vary from glabrous to densely short-pubescent with the hairs acute or at times minutely gland-tipped. A large corolla is also rather characteristic, the tube averag- ing 20 mm. long (range 17 to 23 mm.), and the broadly to narrowly obovate lobes somewhat over half as long. Its color is almost always a brilliant phlox-purple, and pallid or albino forms are decidedly rare. The styles range in length from 12 to 20 mm., bringing the stigmas either into the midst of the anthers, or slightly exserted beyond them. Cultivation. — A Phlox with showy flowers such as charac- terize this species could not fail to early attract the attention of horticulturalists, and according to Sims^ it was introduced into English gardens before 1801. In subsequent years it has been occasionally offered by dealers, though not always under the same name. The confusion of the two species, P. ovata and P. Carolina, already mentioned under History, has resulted in the same material being listed under both these names, or sometimes a combination of the two. Still others have also been applied to strains of this Phlox, such as P. listoniana Sweet^ or P. ovata 3 listoniana Don,^ but the characters are not sufficiently distinctive to justify this. In rich garden soils this species is not very long lived, but it is well suited for wild areas where the soil is sterile and acid, there yielding a bril- liant display. 1 Curtis 's Botanical Magazine 15: pi. 528. 1801. 2 Hortus Brittanicus, ed. 2. 368. 1830. 3 Gen. Hist. Dichl. Plants 4: 241. 1838. I' 30 PROCEEDINGS OF THE Bartonia, No. 13 Plate 7 I h;i«! ■t 11. Phlox Carolina Linne. Thick-leaf Phlox. Plate 7. History.— 0^ all our eastern species of Phlox, the present one has proved the most difficult to interpret. Few current manuals or floras so much as mention it, and those which do combine it with either the next-preceding or the next-following species. That it deserves independent recognition, however, seems evident from the data here assembled. The first known account of this Phlox was that published, accompanied by a colored plate, by Martyn^ in 1728; he termed it ' ' Lychnidea Caroliniana, Floribus quasi umbellatim dispositis ; f oliis lucidis, crassis, acutis. ' ' Four years later it was figured by Dillenius^ as ''Lychnidea folio salicino/' Linne overlooked these articles in preparing the first edition of his Species Plantarum, but evidently obtained a specimen shortly thereafter, for he included it in the second edition,^ with a discussion of its differences from P. glaherrima. During the following fifty years a number of other names were proposed for this same Phlox. In the eighth volume of his Vegetable System, dated 1773, HilP included it as the ''Rugged-stalked Brightweed, P. caroliniana/' this form of the name apparently representing a misprint for Carolina, as it had been spelled by Linne. In 1794 P. altissima was sug- gested by Moench^ as a substitute name. Nine years later Michaux^ described a P. triflora from the uplands of Carolina, and a photograph' of his specimen shows that he had the same species. Plants collected by Fraser had meanwhile been suc- cessfully cultivated at Malmaison, and in 1804 Ventenat,« unaware of the existence of all these prior names, added still another, P. suffruticosa; and this has been extensively used in horticultural writings, both for the present species and for apparent hybrids between it and P. maculata. 1 Hist. PI. Ear. 10, pi. 10. 1728 ; ed. 2. 4, pi. 9. 1752. 2Hortus Elthamensis 1: 205, pi. 166, f. 203. 1732. 3 Species Plantarum, ed. 2. 1 : 216. 1762. 4 Vegetable System 8: 32, pi. 31, fig. 4. 1773. sMethodus Plantas Hort. Marburg. 454. 1794. 6 Flora Boreali- Americana 1 : 143. 1803. 7 Obtained through the kindness of Mr. Ivar Tidestrom. 8 Jardin Malmaison 2 : no. 107, note. 1804. Fig. 1. Phlox Carolina triflora. South of Clinton, Anderson County, Tennessee. Fig. 2. P. c. heferophylla. Cult.; from Alabama. Fig. 3. P. c. altissima. Cult. ; from North Carolina. 30 PROCEEDINGS OF THE Bartonia, No. 13 Plate 7 I !^; 11. Phlox Carolina Linne. Thick-leaf Phlox. Plate 7. History.— Oi all our eastern species of Phlox, the present one has proved the most difficult to interpret. Few current manuals or floras so much as mention it, and those which do combine it with either the next-preceding or the next-following species. That it deserves independent recognition, however, seems evident from the data here assembled. The first known account of this Phlox was that published, accompanied by a colored plate, by Martyn^ in 1728; he termed it ' ' Lychnidea Caroliniana, Floribus quasi umbellatim dispositis; foliis lucidis, crassis, acutis.'' Four years later it was figured by Dillenius" as ** Lychnidea folio salicino.'' Linne overlooked these articles in preparing the first edition of his Species Plantarum, but evidently obtained a specimen shortly thereafter, for he included it in the second edition,^ with a discussion of its differences from P. glaherrima. During the following fifty years a number of other names were proposed for this same Phlox. In the eighth volume of his Vegetable System, dated 1773, HilP included it as the *' Rugged-stalked Brightweed, P. caroliniana/' this form of the name apparently representing a misprint for Carolina, as it had been spelled by Linne. In 1794 P. altissima was sug- gested by Moench^ as a substitute name. Nine years later Michaux^ described a P. triflora from the uplands of Carolina, and a photograph^ of his specimen shows that he had the same species. Plants collected by Fraser had meanwhile been suc- cessfully cultivated at Malmaison, and in 1804 Ventenat,^ unaware of the existence of all these prior names, added still another, P. suffniticosa; and this has been extensively used in horticultural writings, both for the present species and for apparent hybrids between it and P. maculata. 1 Hist. PI. Ear. 10, pi. 10. 1728; ed. 2. 4, pi. 9. 1752. 2Hortus Elthamensis 1: 205, pi. 166, f. 203. 1732. 3 Species Plantarum, ed. 2. 1: 216. 1762. 4 Vegetable System 8: 32, pi. 31, fig. 4. 1773. 5 Methodus Plantas Hort. Marburg. 454. 1794. 6 Flora Boreali- Americana 1: 143. 1803. 7 Obtained through the kindness of Mr. Ivar Tidestrom. 8 Jardin Malmaison 2: no. 107, note. 1804. Fig. 1. Phlox Carolina trifJora. South of Clinton, Anderson County, Tennosseo. Fig. 2. P. c. hctcrophifUa. Cult.; froui Alabama. Fig. 3. P. c. altissima. Cult.; from Xortli Carolina. 1 ! 1 i PHILADELPHIA BOTANICAL CLUB 31 In the first volume of his Flora, Pursh^ included P. Carolina, with P. triflora as a doubtful synonym; then in the supple- ment at the end of the second volume^* he added a P. nitida, noting that it ** approaches near to P. Carolina/' but flowers later and longer. Elliott^ followed Pursh in listing both names, remarking under P. nitida that ''this has generally been considered in this country as the P. Carolina. . . . Lin- naeus however may have united two species under his P. Caro- lina.'' P. carnea Sims,^ supposedly related to P. suaveolens, and P. revoluta Aikin* also differ from P. Carolina Linne in only minor particulars. When Bentham'^ came to monograph the Phloxes, he recog- nized that several of the names above cited referred to the same species, but he considered partial separation possible, and accordingly listed under P. Carolina three varieties; a ovata, 3 nitida, (including P. suffruticosa and approaching P. glaher- rima), and y puberula (replacing P. triflora). The first of these may be withdrawn from consideration here because it represented a distinct species, already discussed ; how the other two should be treated can best be decided after the variability of P. Carolina has been described. Recognizing that the Linnean name ovata had priority over Carolina, Gray^ preferred the former as a species designation, and made the original P. Carolina a variety, P. ovata elatior. Later,^ however, he reduced it to complete synonymy with P. ovata, considering it distinguishable only as ''a taller form, with narrower more tapering leaves and pointed calyx-teeth, approaching the next species [P. glaherrima].^^ At the same time he classed the Ventenat plant as P. glaherrima var. suffru- ticosa, with several of the names above listed as synonyms of it. One additional name, P. heterophylla Beauvais, was pub- lished by Brand.^ 1 Flora Americae Sept. 1 : 149. 1814 ; la 2 : 730. 1814. 2 Sketch Botany S. C. and Ga. 1: 245 [1821]. 3 Curtis 's Botanical Magazine 47: pi. 2155. 1820. 4 Trans. Md. Acad. Sci. Let. 1 : 90. 1837. 5 In De CandoUe's Prodromus 9: 304. 1845. eProc. Amer. Acad. Arts Sci. 8: 249. 1870. 7 Syn. Flora N. A. 2, pt. 1 : 130. 1878. 8 In Engler 's Pflanzenreich 4. 250 : 65. 1907. i i V 41 \ < t \ > 32 PROCEEDINGS OF THE Geography. — Accepting Phlox Carolina as comprising the plant so named by Linne and the various intermediates be- tween P. ovata, P. glaherrima, and P. maculata to which so many other names have been given, its range is as follows: Chiefly in the Blue Eidge and Appalachians, from Georgia and Alabama to Maryland, with local extensions into the Piedmont, Central Lowland, and Coastal Plain. Fio. 4. Distribution of Phlox Carolina. Alabama; Wide-spread in this state, having been collected in 20 counties : Barbour, Choctaw, Cullman, Dallas, De Kalb, Elmore, Etowah^, Jefferson', Lawrence, Lee, Lowndes^ Macon, Marengo, Monroe, Montgomery, St. Clair^ Sumter^ Tusca- loosa, Walker^ and Wilcox^ Both unusually large and un- usually small-leaved forms occur. Florida : Specimens in herbaria from several places in Gads- den and Washington Counties appear to represent this species. Georgia: Occasional in the Blue Ridge and Piedmont, in Catoosa, Cobb, Dade, De Kalb, Fannin, Floyd, Gwinnett, Haralson, Macon, Pickens, Rabun', Richmond, and Spalding counties. Indiana: The northern variety enters this state, as shown by collections made in Crawford, Perry, and Posey counties by Mr. C. C. Deam. PHILADELPHIA BOTANICAL CLUB 33 Kentucky: Scattered through the eastern uplands, speci- mens having been seen from Bath, Breathitt, Estill, Lyon, Marshall, and Powell counties. Maryland : In 1837 Aikin^ recorded from this state a Phlox revoluta, his description of it corresponding closely to the variety of P. Carolina which ranges farthest northeastward. He apparently obtained it in Frederick County, although it has not subsequently been collected there. Mississippi : Material supposed to represent P. glaherrima, from Jackson, Noxubee, and Scott counties, shows such long sepals that it is here regarded as P. Carolina instead. North Carolina: The specific name of this Phlox is very appropriate, for at least two varieties are wide-spread and fre- quent in the Blue Ridge and Piedmont of this state, the county list being: Alleghany^, Buncombe, Burke, Caldwell, Forsyth, Haywood, Henderson', Iredell, Jackson', Lincoln, McDowell', Macon', Madison, Mitchell, Polk', Rowan, Swain, and Tran- sylvania'. Ohio : In the Lea Herbarium, preserved at the Academy of Natural Sciences of Philadelphia, there is a specimen of the northern variety collected in 1839 in the vicinity of Cincin- nati, Hamilton County. It has subsequently vanished there, but has recently been discovered in Adams County by Miss E. Lucy Braun [too late to enter on map]. South Carolina: Not infrequent in the Blue Ridge and Piedmont, in the counties : Abbeville, Aiken', Anderson, Green- ville', Oconee, Pickens, and Richland. The name Phlox nitida was applied by Elliott^ to material from the last. Tennessee : Occasional in the eastern part, specimens hav- ing been seen from Anderson', Claiborne, Cocke, Franklin, Grainger', Hamilton, Haywood, Knox, Madison, Monroe, Polk', Roane, and Robertson counties. Most of these represent the northern variety. Virginia : Known at present only south westward, the county list being: Alleghany', Bath', Bedford, Carroll', Pittsylvania, Rockbridge, and Wythe. 1 Trans. Md. Acad. Sci. Let. 1 : 90. 1837. 2 Sketch Botany S. C. and Ga. 1: 245 [1821]. 34 PROCEEDINGS OF THE Ecology.— The habitat relations of this species are essen- tially the same as those of the next preceding one, P. ovata, the best development of both occurring in subacid soil in open woodlands of intermediate successional stages. When all its varieties are taken into consideration, however, the blooming period of P. Carolina is much longer, extending from early May to late September. Some colonies produce flowers prac- tically throughout the season, others only during one or two of these months. Variation.— The chief reason for the confusion existing as to the status of Phlox Carolina lies is its extraordinary varia- bility in habit and foliage, with seeming gradations toward three or four other species. The existence of numerous inter- mediates between its extreme forms, however, favors its treat- ment as a single species with several varieties. Both flowering and sterile shoots are usually erect, attaining a height of 40 to 150 cm. Occasionally a sterile stem will become decumbent, but the prostrate portion is then much longer, and the leaves on the ascending portion are smaller, than in the corresponding shoots of P. ovata. The number of nodes below the inflorescence is rarely as few as 5, frequently 7 to 11, and occasionally as many as 15 or even 25. The stems are often red-mottled, especially in fall-blooming forms, and the red pigment may extend into the leaves and calyces, making them bronzy green when fresh, and dull brownish when dried. Pubescence is better developed than on any of the related species, the stem being covered with short stiff hairs, or sometimes with long soft ones ; these usually ascend well up the petioles and pedicels, but stop abruptly at the leaf- blades and sepals, rarely spreading also over these. The lowest leaves always have broad petioles which widen gradually into linear or narrowly elliptic blades, and succes- sively higher ones normally lose the petioles and attain greater blade-width. In some colonies occasional individuals, and in others the majority of the plants, may have the uppermost leaves little broader than linear-elliptic, the aspect being .then much like that of P. glaherrima. Usually, however, the blades of the upper leaves become elliptic- or oblong-lanceolate to PHILADELPHIA BOTANICAL CLUB 35 ovate. Locally, especially southwestward, the uppermost leaves may be even more broadly ovate and large-sized than those of P. ovata itself. Phlox Carolina, in most of its occurrences, tends to develop a corymbose inflorescence, the outer branches rapidly elon- gating until they much exceed their subtending bracts. On individual plants, or sometimes in whole colonies, however, these branches remain short, resulting in a broadly or even narrowly conical panicle. Here the gradation is toward P. maculata, in which the inflorescence is normally cylindric, though occasionally it becomes conical instead. In the section key the length of the calyx of P. Carolina was given as 6 to 11 mm., but this whole range is not met with in any one colony or even within one variety. A separation can be made, in fact, into three length-classes : the calices of north- ern spring-blooming plants are longest, 8 to 11 mm. ; those of southwestern spring-blooming ones intermediate, 7.5 to 9.5 mm. ; and those of southeastern fall-bloomers shortest, 6 to 9 mm. The first of these classes resembles in this respect P. ovata, the second P. glaherrima, and the third P. maculata. Other calyx characters, such as extent of union of sepals (relative length of tube and lobes), width of membranous intercostae, etc., seem to vary erratically. Less variability is shown in corolla-characters. The bril- liant phlox-purple color gives way to pale or albino phases somewhat more frequently than in P. ovata. The tube and lobes average a little smaller than in the latter, but the range in size is proportionately about the same. The style is usually nearly as long as the corolla-tube, with the anthers lying close to the stigmas, and shows only negligible variation. Although extreme variants often look like distinct species, so many intergradations occur between them that no key can be drawn up for the identification of isolated specimens, and the species is accordingly here divided into varieties as has been done with several members of sections previously dis- cussed. 36 PROCEEDINGS OP THE PHILADELPHIA BOTANICAL CLUB m \ 37 S| VAEIETIES AND FOEMS OF PHLOX CAROLINA Nodes about 8; upper leaves elliptic to ovate-lanceolate; calyx 8-11, AVERAGING 9.5 MM. LONG; BLOOMING EARLY. Phlox Carolina triflora (Michaux) Wherry, status novus. P. triflora Michaux. P. carnea Sims. P. revoluta Aikin. P. Carolina y pulerula Bentham. P. ovata elatior Gray. N. C. to Ind. and Md. Forms : Pubescent. Stem and leaves bearing long soft hairs. Typified by speci- men in Academy of Natural Sciences of Philadelphia from Biltmore, Buncombe Co., N. C, Biltmore Herbarium 706a, June 21, 1897. Rare. Narrow-leaved. Even uppermost leaves linear-lanceolate. Typitied by specimen from open woods 3 miles north of Millspring, Polk Co., N. C collected by E. T. W. May 27, 1929. Occasional throughout. Nodes about 8; upper leaves often broad-elliptic to ovate; CALYX 7.5-9.5, AVERAGING 8.5 MM. long; blooming EARLY. Phlox Carolina heterophylla (Beau- vais) Wherry, status novus. P. heterophylla Beauvais ex Brand. P. carnea Brand, not Sims. Wide-spread in Ala. and occa- sional in adjacent portions of Fla., Ga., and Miss. Forms: Pubescent. Stem and leaves bear- ing long soft hairs. Typified by specimen from woods 6 m. south- west of Devenport, Lowndes Co., Ala., collected by E. T. W. June 3, 1927. Occasional. Pallid. Corolla pink. Typified by specimen from open woods 1 mile east of Oakman, Walker Co., Ala., collected by E. T. W. May 8, 1929. Rather rare. The morphology and distribution of the above listed varie- ties suggest their evolutionary relations to have been somewhat as follows : P. c. heterophylla first developed in the mountains of Alabama, and its offspring spread in various directions. One descendant proved adapted to conditions farther north, and gave rise to the variety P. c. triflora; it evidently arose from a colony showing tendencies toward moderately narrow leaves and unusually long sepals. Another was more success- ful at higher elevations, developing into P. c. altissima with Nodes about 12 (up to 25) ; up- per LEAVES often ELLIPTIC TO oblong-lanceolate ; CALYX 6-9, AVERAGING 7.5 MM. LONG; blooming LATE. Phlox Carolina altissima (Moench) Wherry, status novus. P. caro- liniana Hill. P. altissima Moench. P. suffruticosa Vent. P. nitida Pursh. P. Carolina P nitida Benth. P. maculata ni- tida Chapm. P. glaherrima suf- fruticosa Gray. N. Ga., E. Tenn., and N. C. Forms: [Though decidedly variable in stature, foliage, etc., this va- riety is not readily separable into definite forms]. The type specimen of P. Carolina in the Linnean herbarium corre- sponds most closely to this variety. increased stature, shorter calyx, and later blooming season. Still others became so much further differentiated from the parent stock that they are now considered distinct species. Cultivation. — Phlox Carolina had been introduced into hor- ticulture before 1728, for in that year Martyn^ recorded its being grown in Cowell 's nursery at Hoxton. It was also listed by Dillenius^ in 1732 among the plants in the Eltham garden. The original stock seems to have died out, however, for the material cultivated at Malmaison in 1804, named by Ventenat^ P. suffruticosa, represented a re-introduction by Fraser; and the figures of P. Carolina published by Sims* in 1810, and of P. carnea which he^ proposed as a new species ten years later, were also stated to be based on Fraser plants. Two of the supposedly distinct species pictured by Maund^ and Sweet^ shortly afterwards, P. triflora and P. penduliflora, appear to be merely cultivated forms of the original native P. Carolina, For some obscure reason the term suffruticosa came to be preferred over any of the others, and is today generally applied in horticultural writings to the species under discus- sion. In ** Standardized Plant Names'' Olmsted, Coville, and Kelsey^ listed nine named forms of this in the American trade ; the most widely known of these is **Miss Lingard," a sterile clone which probably originated from a crossing of two of the native varieties. Should horticulturalists come to follow the same code of nomenclature as botanists, however, the earliest name, Carolina, will have to be used as the specific designation. The remaining two species of the section Ovatae, together with those of the section Paniculatae, will be discussed in a future number of Bartonia. iHist. PI. Rar. 10, pi. 10. 1728; ed. 2. 4, pi. 9. 1752. 2Hortus Elthamensis 1: 205, pi. 166, f. 203. 1732. 3 Jardin Malmaison 2 : no. 107, note. 1804. * Curtis 's Botanical Magazine 33: pi. 1344. 1810. 5 Curtis 's Botanical Magazine 47: pi. 2155. 1820. 6 Botanic Garden 1 : no. 6. 1825. 7 British Flower Garden 1 : no. 29. 1823 ; ser. 2. 1 : no. 46. 1830. 8 Standardized Plant Names 365. 1923. * •r! I ■ wmi I J ^*"y Reprinted from Bartonia, No. 14, 1932 Bartonia, No. 14 Plate 2 The Eastern Long-styled Phloxes, part 2^ Edgar T. Wherry 12. Phlox glaberrima Linne. Smooth Phlox. Plate 2. History. — The earliest known reference to this Phlox was that published by Plukenet^ in 1705, his description being **Lychnidea Asclepiadis folio Floridana, summo caule flori- bunda. ' ' It was first figured by Dillenius,^ with the charac- terization **Lychnidea folio melampyri. " In establishing the genus Phlox in 1737, Linne* evidently had this species pri- marily in mind, for it is the only one he listed in the catalog of the plants in Clifford's garden.^ Where these cultivated plants came from is unknown, but it may well have been east- ern Virginia, Gronovius^ having recorded a ** Phlox foliis lineari-lanceolatis " there in 1739. The name Phlox glaber- rima was applied to it by Linne^ in the Species Plantarum, and has been used by practically all subsequent writers, although Salisbury^ preferred P. melampyrifolia. As already pointed out under P. Carolina, that species has been combined with the present one by many botanists, cer- tain forms of it being indeed classed by Gray® as P. glaber- rima var. suffruticosa. The reinstatement of P. Carolina, how- ever, calls for a transfer of this variety to it. Finally, in his monograph of the Polemoniaceae, Brand^® described as P. glaberrima subvar. angustissima a small-leaved variant from the Gulf Coastal Plain. 1 Contribution from the Botanical Laboratory of the University of Pennsylvania. Part 1 appeared in Bartonia 13: 18. 1932. 2 Amaltheum Botanicum 136. 1705. 3 Hortus Elthamensis 1 : 203, pi. 166, f . 202. 1732. * Grenera Plantarum 52. 1737. 5 Hortus Cliffortianus 63. 1737. « Flora Virginica 21. 1739. 7 Species Plantarum (1) : 152. 1753. 8 Prodromus Stirpium 123. 1796. »Syn. Flora N. A. 2, pt. 1: 130. 1878. 10 In Engler^s Pflanzenreich IV. 250: 64-65. 1907. (14) Fig. 1. Phlox glaberrima melampyrifoUa. Northeast of Williston, Barnwell County, South Carolina. Fig. 2. Phlox glaherrima i7iterior. West of Wheatland, Knox County, Indiana. f ^1 Reprinted from Bartonia, No. 14, 1932 Bartonia, No. 14 Plate 2 The Eastern Long-styled Phloxes, part 2' Edgar T. Wherry 12. Phlox glaberrima Linne. Smooth Phlox. Plate 2. History. — The earliest known reference to this Phlox was that published by Plukenet^ in 1705, his description being **Lychnidea Asclepiadis folio Floridana, summo caule flori- bunda.'' It was first figured by Dillenius,^ with the charac- terization *'Lychnidea folio melampyri. ' ' In establishing the genus Phlox in 1737, Linne* evidently had this species pri- marily in mind, for it is the only one he listed in the catalog of the plants in Clifford's garden.^ Where these cultivated plants came from is unknown, but it may well have been east- ern Virginia, Gronovius^ having recorded a ** Phlox foliis lineari-lanceolatis'' there in 1739. The name Phlox glaber- rima was applied to it by Linne^ in the Species Plantarum, and ha^ been used by practically all subsequent writers, although Salisbury^ preferred P. melampyrifolia. As already pointed out under P. Carolina, that species has been combined with the present one by many botanists, cer- tain forms of it being indeed classed by Gray^ as P. glaber- rima var. suffruticosa. The reinstatement of P. Carolina, how- ever, calls for a transfer of this variety to it. Finally, in his monograph of the Polemoniaceae, Brand^^ described as P. glaherrimu subvar. angustissima a small-leaved variant from the Gulf Coastal Plain. 1 Contribution from the Botanical Laboratory of the University of Pennsylvania. Part 1 appeared in Bartonia 13: 18. 1932. 2 Amaltheum Botanicum 136. 1705. sHortus Elthamensis 1: 203, pi. 166, f. 202. 1732. 4 Genera Plantarum 52. 1737. 5 Hortus Cliffortianus 63. 1737. 6 Flora Virginica 21. 1739. 7 Species Plantarum (1): 152. 1753. sProdromus Stirpium 123. 1796. »Syn. Flora N. A. 2, pt. 1: 130. 1878. 10 In Engler's Pflanzenreich IV. 250: 64-65. 1907. (14) Fuj. 1. Phlox (jlabt'irhna mcUunpiiri folia. Northeast of Willistoii, lianiwoll County, South Cjirolina. Fig. 2. Phlo.r r/lahrrrinia inierior. West of Wheatland, Knox County, Indiana. INTENTIONAL SECOND EXPOSURE II '!Ei ^ |1| 1-y » I ii PHILADELPHIA BOTANICAL CLUB 15 Geography.— The range of this species is decidedly differ- ent from that of the relative with which it has so often been confused : while P. Carolina is essentially an upland plant, P. glaherrima occurs chiefly in the lowlands, the ranges of the two overlapping only in the vicinity of longitude 85° west. As brought out in figure 1, the second-named species extends from northern Florida to southeajstern Virginia and to east- ern Texas, and thence northward to southeastern Wisconsin and northern Ohio. One of its varieties occurs only above the Fall-line (cross-hatched band), the other chiefly below this line, though locally entering adjacent provinces. Fig. 1. Distribution of Phlox glaherrima. Alabama: Occurs practically throughout, the county list being: Autauga, Bibb, Cherokee', Chilton', Clarke', Coffee', Coosa', Cullman, Dallas, Elmore, Etowah',. Fayette', Geneva', Lee, Limestone', Marengo', Marion', Mobile, Montgomery, Rus- sell', St. Clair, Talladega', Tuscaloosa', Walker', and Wash- ington'. Arkansas: Occasional southward and eastward, specimens being preserved from Arkansas, Clark, Craighead, Faulkner, Hot Spring, Jefferson, Nevada, Prairie, and Pulaski counties. Florida: Known from several places in Gadsden' County. 16 PROCEEDINGS OF THE Georgia : Frequent, except toward the northeast and south- east corners, the county list being: BartoV, Bibb', Bryan Bulloch, Calhoun', Dade, Early', Emanuel', Floyd, Gilmer', Jefferson', Jenkins', Laurens', Liberty, McDuffie, Montgom- ery' Muscogee, Peach, Randolph', Richmond, Spalding, Tatt- nall', Telfair', Troup', Twiggs', Washington', and Wheeler'. Illinois : The northern variety occurs practically through- out specimens having been seen from 24 counties: Cass, Champaign, Christian, Cook, Du Page, Jackson, Kankakee, Knox, Lake, La Salle, Lee, Livingston, Macon, Macoupin, Marion', Menard, Peoria, Randolph, Richland, St. Clair, Sangamon, Wabash, Will, and Woodford. Indiana : Known from even more counties than in the next- preceding state, chiefly owing to the eoUecting activity of Mr. Charles C. Beam : Bartholomew, Benton, Clark, Clay, Craw- ford Daviess, Dubois, Fulton, Gibson, Harrison, Jasper, Jay, Jefferson, Jennings, Knox', Lake, Laporte, Newton, Owen, Porter Posey, Pulaski, Putnam, Shelby, Spencer, Starke, Sullivan, Tippecanoe, Vanderburg, Vermilion, Vigo, Warrick, Washington, and White. [Iowa: Included by W. Greene,^ apparently by mistake.] Kentucky: As little collecting has been done, only 8 county records exist: Bell, Daviess, Edmonson, Henderson, Logan, McCracken, Marshall, and Powell. Louisiana: Rare, specimens having been seen only from Bienville and Rapides parishes. [Minnesota: Reported by MacMiUan^ from "Forest dis- trict to New Ulm," but Prof. C. 0. Rosendahl advises me that there are no specimens in the State University herbarium, glabrate forms of P. pUosa having perhaps been taken for it. J Mississippi: Fairly common, the county list being: Clarke, Greene', Grenada, Harrison, Jackson, Jones, Lafayette^ Madi- son, Noxubee, Oktibbeha, Scott, Tishomingo, and Wayne. Subvar. angustissima Brand' was from Biloxi, which is m Harrison County in this state, and not Missouri, as given. 1 Plants of Iowa 206. 1907. 2 Metaspennae Minn. Valley 432. 1892. sin Engler's Pflanzenreich IV. 250: 65. 1907. PHILADELPHIA BOTANICAL CLUB 17 Missouri: Common southeastward, in the counties: Bol- linger, Butler, Dunklin, Jefferson, Ripley, St. Louis, Scott, and Wayne ; reports further west are unconfirmed. North Carolina : Apparently rare, specimens being in the herbaria studied only from Craven, Randolph, and Rowan counties. Records from the Blue Ridge represent P. Carolina instead. Ohio: Occasional toward the west side, in Butler, High- land, Lucas, and Wood counties ; elsewhere other species have been mistaken for it. South Carolina: Frequent southward, the county list being: Aiken', Barnwell', Beaufort', Berkeley, Charleston, Dorchester, Florence', Hampton, Jasper, and Lexington'. Tennessee : Owing to lack of collecting, specimens are pre- served only from Coffee, Davidson, Dickson, Haywood, and Madison counties. Texas : A vigorous form has been collected at two localities in Smith County. Virginia : In proposing the name P. glalerrima, Linne re- ferred to Gronovius's record of a Clayton plant, and as the latter collected chiefly in Gloucester County, Virginia, the species may be presumed to have formerly grown there. It is not known to have been found in subsequent times, however, having probably been exterminated by agriculture. Wisconsin: Limited to the southeast corner, there being records for Kenosha and Racine counties only. Ecology.— While both varieties of Phlox glaberrima grow in rather moist places, the northern one is usually found in circumneutral soil in grassy meadows, the southern in decid- edly acid soils in a variety of habitats, including pinelands, pitcher-plant meadows, thickets along brooks, and even nch woods on river flood-plains. In all cases the associations ap- pear to be climax or subclimax ones for the areas. The bloom- ing period begins in late Spring and extends to mid-Summer or occasionally to early Autumn. Insect visitors observed by Robertson' comprised 7 butterflies and one fly. 1 Ecology 8 : 119. 1927 ; Flowers and Insects 151. 1928. 18 PROCEEDINGS OF THE Variation.— Phlox gUherrima is a markedly variable spe- cies, especially in habit and foliage characters. Differences in the availability of moisture or of nutrient elements, as well as inherited tendencies, have been observed to lead to a range in height from (25-) 50 to 100 (-175) cm. The leaf-outline is normally linear to lanceolate or oblong-lanceolate, and only exceptionally approaches the elliptic shape so frequent in P. Carolina; the larger leaves are from (35-) 50 to 150 (-200) mm. long, and (3-) 5 to 15 (-25) mm. wide. Although as the name implies the plants are sometimes wholly glabrous, a certain amount of pubescence is often present. Most fre- quently this appears on the inner surface of the sepals and on the pedicels, but it may extend down over the bracts, stems, and even the leaves. This tendency seems to reach its maximum along the Gulf coast, in the very region where P. pilosa shows diminished pubescence. In the key to the species of section Ovatae in the preceding article in this series, the limits of calyx-length for P. glaher- rima were stated, in round numbers, as * ' 6 to 8 mm. ' ' Actu- ally the limits are slightly wider than this when extremes are taken into account, and two geographic varieties are recog- nizable on the basis of length and extent of union of the sepals. In the southern one these range from 6.5 to 8 (-8.5) mm. long, and are united to about half their length ; in the northern, from (5.5-) 6 to 7.5 mm., with the union extending well beyond the middle.^ The corolla-color is usually phlox-purple, with occasional gradation toward a slightly bluer or redder hue. Pallid and albino mutants appear in small numbers in most large colo- nies. The pigment may be uniformly distributed over the corolla-face, or may diminish toward the center, forming a pale eye bearing a more or less distinct five-rayed star of deeper shade. In dimensions the corolla varies moderately: the tube-length from (16-) 18 to 23 (-25) mm., the lobes from about 7 x 5 to 10 x 9 mm. The styles range from (11-) 15 to 20 (-22) mm. in length, occasionally becoming exserted. 1 The measurements given have been made on dried specimens, in which some shrinkage often occurs, so that fresh material may yield slightly greater values. PHILADELPHIA BOTANICAL CLUB 19 VARIETIES AND FORMS OF PHLOX GLABERRIMA Sepals 5.5-7.5 mm. long, united to about % their length ; calyx -lobes thus 1.5-3 mm. long. Phlox glaberrima interior Wherry, var. nov. Sepala 5.5 ad 7.5 mm. longa, circa % longitudine conjuncta. Named from the region of its occurrence. Typified by specimen from 2 miles west of Wheatland, Knox Co., Ind., collected by E. T. W. June 5, 1931. Ranges from Ky. to E. Mo., SE. Wise, and N. Ohio. Forms : Maculate-stemmed. Typified by specimen from 7 miles southwest of Boonville, Warrick Co., Ind., collected by C. C. Deam June 5, 1930, in herbarium of E. T. W. Occasional throughout. Pubescent-pedicelled. Typified by specimen in U. S. National Herbarium from Jackson Co., 111., collected by G. H. French July 17, 1873. Rare. Light-colored (pink, white, etc.) Typified by specimen in Field Museum herbarium from Ravenswood, Cook Co., 111., collected by R. N. Lloyd Aug. 15, 1887. Infrequent. Sepals 6.5-8.5 mm. long, united to V2-% theie length ; calyx-lobes thus 2.5-4 mm. long. Phlox glaberrima melampyrifolia (Salisbury) Wherry, status novus. P. glaberrima Linne. P. melampyrifolia Salisbury. Ranges from NW. Fla. to E. Tex., SW. Mo., and SE. Va., at low elevations. Forms : Small-leaved. P. g. subvar. angustissima Brand. Common in sterile situations, such as pitcher-plant meadows. Maculate-stemmed. Typified by specimen from 4^ miles north of Samantha, Tuscaloosa Co., Ala., collected by E. T. W. May 8, 1929. Frequent. Pubescent. Typified by specimen from 21 miles west of Jackson, Clarke Co., Ala., collected by E. T. W. July 12, 1932. Most often observed in Ala. and Miss., but may occur elsewhere. Light-colored (pink, white, etc.) Typified by specimen from 2 miles northwest of Fletcher, Chilton Co., Ala., collected by E. T. W. July 14, 1932. Rather rare. Cultivation.— As noted under History, the early records of this Phlox referred to cultivated material, which was prob- ably sent to Europe from eastern Virginia. When trans- planted from the wild into rich garden soil its leaves become larger and its inflorescence more massive, as shown in the plates published, for example, by St. Hilaire,^ Vietz,^ and Sweet.^ It does not become so attractive, however, as does P. Carolina ('*P. suffruticosa'^), and is today rarely seen in gardens. iPlantes France 3: pi. (91). 1809. 2 Abbildung med.-6kon.-techn. Gewachse 8 : pi. 719. 1818. 8 British Flower Garden (2) 1: pi. 36. 1830. Baktonia, No. 14 Plate 3 20 PROCEEDINGS OF THE 13. Phlox macnlata liinn^. Meadow Phlox. Plate 3. His«orj/.-Several pre-Linnean references to this species are known Plukenet^ listed it in 1700 as "Lychnidea Marmna elatiori Alsines aquaticae foliis, floribus in longam spicam dense stipatis," and published a crude but unm^tajable fig- ure of it five years later.^ It was termed by Ray^ Lych- noides Uanlandica Ja^mini fiore quinquepartito, f olus binis oppositis, basi & auriculis caulem utrinque amplexantibus. In the correspondence between Bartram and CoUinson pub- lished by Darlington,* two references to it occur--Collinson to Bartram, June 16, 1742 : "... but I have a Ly ohms, from Doctor Witt, different from any yet that I have seen, it seems to be the king of that tribe. ... It is now about two feet high, and yet no flowers appear. The stalk is most finely spotted,-which is very distinguishing from aU tl^e rest that I have seen."— Bartram to CoUinson, June 11, 1743: Ihe Doctor's famous Lychnis, which thee has dignified so highly, is I think, unworthy of that character. Our swamps and low grounds are full of them. I had so contemptible an opinion of it, as not to think it worth sending ..." In the account of his travels in North America, Kalm= recorded finding it m New Jersey on June 2, 1749, and according to Juel and Harshberger,« a specimen is preserved in his herbarium at Upsala. Finally, it was included in the 1752 edition of Mil- ler's Gardeners' Dictionary,' z& Phlox No. 4. In assigning the species name, Linne« noted that he had received the plant from Kalm, but stated its source as Vir- ginia. As a photograph* of his specimen shows it to represent a variety common only north of that state, and as Kalm did not travel to any extent south of New Jersey, the latter should be regarded as its type locality. 1 Mantissa 122. 1700. 2 Amaltheum Botanieum pi. 425, fig. 6. 17U&. 4 MemorUlfor Bartram aiid MarshaU 156, 164. 1849. 5 Travels into North America 2 : 222. 1771. 6 Proc. Acad. Nat. Sci. Phila. 81 : 303 1929 ■> Gardener's Dictionary, ed. 6. page PH. 175^ : #sf '".♦Cri ^f^^ ■4^r^ Fig. 1. PJilo.r macuhita odoraia. Southeast of Marlton, Burlington County, New Jersey. Fig. 2. PhJox maculaUi p}iramulaJ'\><. \\\ cultivation; originally froui Maryland. PHILADELPHIA BOTANICAL CLUB 21 Plants which appear to represent varieties or forms of the species, with purple corollas, successively received the names P. maculata var. a purpurea Michaux,^ P. pyramidalis Smith,^ P. odorata Sweet,^ and P. reflexa Sweet.* Albino forms, with the stem scarcely if at all maculate, such as occa- sionally occur in many of its colonies, have been assigned a still larger number of names, including P. suaveolens Aiton,*^ P. alha Moench,^ P. maculata var. (3 Candida Michaux,^ P. tardiflora Penny,« P. longiflora Sweet,^ P. omniflora Loudon,^^ and P. maculata var. suaveolens (Alton) Brand.^^ Several additional names in the literature probably apply to this species, but the descriptions accompanying them are too in- complete for certain identification, and type specimens are lacking. Geography,— On the map (Fig. 2) which appears on the following page, the range of Phlox maculata is indicated as fully as available information will permit. Although it has been reported at one time or another from nearly all the southern states, no specimens certainly referable to it have been seen from south of latitude 35°, and P. Carolina or P. glahernma, both of which at times have maculate stems, ap- pear to have been mistaken for it. Unlike the next-preceding species, its distribution shows no evident relation to any geo- logical or topographic boundaries. The range as a whole extends from central North Carolina to eastern Missouri, southeastern Minnesota, and southern Quebec, with garden escapes eastward into New England. It thus shows the least tendency to push southward of all the eastern species. 1 Flora Boreali- Americana 1 : 143. 1803. 2 Exotic Botany 2: 55, pi. 87. 1806. 3 British Flower Garden 3 : pi. 224. 1827. 4 British Flower Garden 3 : pi. 232. 1827. 5 Hortus Kewensis 1 : 206. 1789. 6 Supplementum Plantarum 173. 1802. 7 Flora Boreali- Americana 1 : 143. 1803. 8 Hortus Epsomensis 38. 1828. 9 British Flower Garden 4: pi. 31. 1830. 10 Hortus Brittanicus, Suppl. 2: 656. 1839. 11 In Engler's Pflanzenreich IV. 250: 60. 1907. 22 PROCEEDINGS OF THE [Alabama : Mohr^ recorded this species from four counties, and also stated it to be '* abundant on the Warrior table- land/' but his specimens, preserved in the U. S. National Her- barium, all represent P. Carolina.] [Arkansas : Included by Branner and Coville^ in their list, but no specimens are present in the herbaria seen.] Connecticut: Known in two counties, Fairfield, where it may be native, and New London*, where it has escaped. Delaware : Common in New Castle^ County. District of Columbia' : Both varieties once occurred here. Fig. 2. Distribution of Phlox maculata, Illinois: Apparently rather rare, the county list being small: Champaign, La Salle, Lee, Piatt, Stark, Tazewell, Ver- milion. Will, and Woodford. Indiana : Kecorded from 28 counties, although Mr. Charles C. Deam advises me that he does not consider it as really com- mon: Cass, Clark, Decatur, Delaware, Fayette, Gibson, Ham- ilton, Henry, Jackson, Jay, Jefferson, Jennings, Lake, Mar- shall, Miami, Monroe, Montgomery, Posey, Randolph, Ripley, St. Joseph, Scott, Shelby, Tippecanoe, Vigo, Washington, Wayne, and White. 1 Plant Life of Ala. 685. 1901. 2 Ann. Eept. Geol. Survey Ark. 1888: 204. 1891. PHILADELPHIA BOTANICAL CLUB 23 lowA: Frequent in the eastern part, specimens or records having been seen from the counties: Black Hawk, Cerro Gordo, Clinton, Fayette, Floyd, Grundy, Hancock, Henry, Johnson, Jones, Linn, Poweshiek, Story, Winneshiek, and Wright. Kentucky: Both northern and southern varieties are known here, the latter being represented by an unusually narrow-leaved form. Specimens are preserved from 8 coun- ties: Bell, Clark, Fayette, Henderson, Lincoln^ McCreary', Owen, and Rockcastle, although it probably occurs in many others also. Maryland: Common in the central and western portions, the two varieties intermingling ; the county list is : Allegany, Baltimore, Cecil^ Frederick, Garrett, Harford^, Howard, Montgomery^, Prince Georges', and St. Marys^ [Massachusetts: Escaped from cultivation in Berkshire' and Bristol' counties.] Minnesota: Extends into the southeastern counties, her- baria containing specimens from Dakota, Dodge, Goodhue, Mower, Olmsted, and Wabasha. [Mississippi : Three records published by Lowe^ are believed to represent maculate-stemmed forms of P. glaberrima,] Missouri: Though sometimes stated to be common in this state, the species must actually be very rare there, as the only specimens seen in all the herbaria examined are from Iron County. New Jersey : The northern variety is abundant throughout, except in the pine-barrens, there being 11 county records: Burlington^ Camden, Cape May^ Gloucester, Hunterdon, Mercer, Middlesex, Monmouth, Ocean, Passaic, and Salem. The material on which the species was founded had been col- lected here by Kalm. New York : Rather rare, only 8 counties being represented by specimens, and some of these are considered by House^ as escapes : Broome, Cattaraugus, Erie, Essex', Hamilton, Lewis, Rensselaer', and Saratoga*. 1 Plants of Miss. 233. 1921. 2 Ann. List Ferns & Fig. Pits. N. Y. St. 580. 1924. 24 PROCEEDINGS OF THE PHII/ADEIiPHIA BOTANICAL CLUB 25 North Carolina : Although many reporte of this species here really represent P. carolim, the southern P. macMa does occur in at least 13 counties: Avery, Buncombe, Chat- ham, Durham', Forsyth, Guilford, Haywood, Iredell, Jax^k- son' Madison, Orange, Randolph, and Yancey . Ohio • The northern variety is common and the southern occasional, in the counties: Adams, Auglaize, Champai^, Clark, Clermont, Clinton, Fairfield, Frankhn', Fulton, Gal- lia, Greene, Hamilton, Hancock, Henry, Hocking, Holmes, Licking, Lucas, Madison, Mahoning, Montgomery, Muskin- gum, Portage, Richland, Scioto, Stark, Summit, Warren, and Wayne. i xi. x * Pennsylvania : The county list is even larger than that f or Ohio: Allegheny, Beaver, Bedford, Be^l^^ Blaij Brad^^^^^^^ Bucks', Butler, Cameron, Center^ Chester^ Clearfield^ Co um- bia Crawford, Cumberland, Delaware^, Payette, Franklin, Huntingdon, Lackawanna, Lancaster^ Lawrence Lehigh, Luzerne, McKean, Mercer, Montgomery, Contour, Nor hamp- ton, Philadelphia, Somerset, Westmoreland^ and YorV. The southern variety enters Lancaster County. , ^ ,, . ^ [South Carolina: Specimens from this state labelled i^. maculata have proved to be P. carolirui instead.] Tennessee: Both the northern and a narrow-leaved form of the southern variety occur here, although there are only 11 county records: Benton, Blount, Carter, Coffee, Knox, Morgan', Putnam, Khea', Roane, Scott', and Wayne. [Vermont : Escaped in Addison' and Windham* counties.] Virginia : Scattered throughout, specimens being preserved from 9 counties: Albemarle, Arlington', Bedford, Fairfax, Floyd', Giles, Grayson', James City, and Smyth. Both varie- ties are well represented. -, • -d West Virginia : Frequent, especiaUy northward, m Bar- bour' Hardy, Pocahontas', Preston, Randolph', Upshur, Web- ster, knd Wirt counties. Unexpectedly, the southern variety appears to be the commoner. Canada Quebec : Extends along the St. Lawrence valley into Brome-Missisquoi, Shefford, and Terrebonne counties. Perhaps in part an escape. Ecology. — Phlox maculata is typically a wet-soil plant, growing in meadows, springy places, and alluvial flats along streams. Locally it may adapt itself to habitats which are moist in Spring but dry out later in the year, then tending to become stunted and bronzed. It prefers moderately acid soil reaction, but occasionally enters both alkaline and mediacid bogs. From the succession standpoint, its associations usually represent the climax for the area, although if forests invade the moist situations, it tends to die out. The northern variety begins to bloom in late Spring, and continues to the middle of Summer; the southern one starts 3 to 6 weeks later, and remains in bloom correspondingly longer. The bright purple, long-tubed, and often decidedly fragrant flowers attract vari- ous kinds of butterflies. Variation. — This species varies greatly in stature, the northern variety ranging from 30 to 90, or rarely to 125 cm., the southern one (40-) 75 to 150 cm. in height. The stem is usually strongly spotted with purple, but sometimes, espe- cially in pale-flowered individuals, is wholly green. Short stiff hairs often cover the stem and inflorescence-branches, but rarely spread over leaf-surfaces. In the northern variety there are from 7 to 15 or exceptionally 20 nodes ; the leaves at the lower ones are linear, but rapidly broaden upward, often becoming ovate with cordate base just below the inflo- rescence. In the southern one the nodes are more numerous and usually more crowded, from 15 to 30 in number, and the leaves widen upward more gradually. In Kentucky and Tennessee even the uppermost leaves are linear-lanceolate, with tapering base, although elsewhere they tend to become oblong-lanceolate, with truncate or subcordate bases. While a cylindrical panicle is characteristic, this varies in length from but 3 to over 30 cm., and branches may develop at the base of the inflorescence, which then takes on a nar- rowly conical shape. In contrast to its relatives, the calyx shows marked uniformity throughout the range, the sepals averaging 7 mm. long and being united to about % their length, and rarely deviating as much as a millimeter from these measurements. 26 PROCEEDINGS OF THE Phlox-purple is the predominating corolla-color, but lighter hues and albinos are occasional. The eye tends to be paler and to bear a more or less distinct purple stripe at the base of each lobe. The tube ranges from 17 to 26 mm. in length, and the lobes from 6 to 11 mm. in both length and width. The styles usually terminate in the midst of the anthers, just below the tube-orifice, but may extend beyond the latter. As noted under History, Michaux proposed two * Varieties'' of P. maculata, but since he was referring only to color-forms, his names are not here adopted. The first name applied to what is now considered a variety was P. pyramidalis of Smith, which is shown by his description to have represented the southern one. Sweet's P. odorata seems to be the especially sweet-scented northern variety, so these are the names given varietal status in the accompanying tabulation. VAEIETIES AND FORMS OF PHLOX MACULATA Nodes pew (about 7-15) and opten remote; upper leaves tending to become broadly lanceolate and cordate. Phlox maculata odorata (Sweet) Wherry, status nevus. P. maculata Jj. P. odorata Sweet. P. reflexa Sweet. Mountains of Tenn. to E. Mo., SE. Minn., and S. Que. Blooms chiefly in early Summer. Form: Light-colored (pink, white, etc.) P. suaveolens Aiton. P. alha Mbench. P. 'macvXata var. suaveolens Brand. Occasional. Nodes numerous (about 15-30), and often crowded; upper LEAVES tending TO BECOME LINEAR- OR OBLONG-LANCEOLATE. Phlox maculata pyramidalis (Smith) Wherry, status novus. P. maculata var. a purpurea Michaux. P. pyramidalis Smith. N. C. to Tenn., S. Ind., and S. Pa. Blooms chiefly in late Summer. Forms : Narrow-leaved. Typified by specimen from Spring City, Ehea Co., Tenn., collected by E. T. W., Sept. 12, 1927. Frequent, Tenn & Ky. Light-colored (pink, white, etc.) P. maculata var. p Candida Michaux. P. tardiflora Penny. P. longiflora Sweet. P. omniflora Loudon. De- velops in most large colonies. Cultivation.— The early introduction of this species into horticulture was referred to under History. That it found favor for a time is indicated by the frequency with which it was illustrated, under one name or another, in the literature of the early 19th century. More recently its use gradually diminished and it is today rarely seen in cultivation. Keprintea fro. Pkoc^os o.JHK P^.v.k. Ac...m, o. Sc^kc, PENNSYLVANIA ACADEMY OF SCIENCE 33 TTPHT nrir AL STUDIES OF SERPENTINE-BABREN ECOLOGKiO^STUmi.^^^ COMPOSITION By Edgar T. Wherry Associcie Professor of Botany, Vni.ersity of Penr^sylvania Many areas classified from the a.ricultu-1^^^^^^^^^^^^ ^-- support a characteristic and of*^'^. l^^^"^'^* ""f l^J^^^^^^^^^^^ particularly true of the serpentine-barrens of eastern ^« J ^hich owe their name to their development over outcrops o ^^.^^^ xnagnesian metamorphic rock, «f P^"*^^^, J.^'J"^ efly discussed by ness of the soils derived from this rock hasbeen briefly Hilgard^ and studied in detail by ^;^^T ^^^^"^^^.^^ .^t with any tural deficiency of these areas ^'^3^°^^.^^° ^/.^^'^^^^^^ supply of the toxic effect of the magnesium ion, but ^I^^^J^^^^^f^^^^^^^^^ m the ordinary nutrient elements, -P-^^^^ P^^^re^h,^^^^^^^^ ability of soils These authors did not discuss, however, the rema certain native plants to thrive on serpentine-barrens. 1 Soils: 36, 1906. 2 Soil Science, 22: 291,1926. In connection with the writer's investigations of plant distribution in the eastern United States, it seemed of interest to obtain data as to the chemical composition of the ash of those growing in such barren situations ; and on recommendation of the Faculty Research Committee, a grant of funds was made by the Board of Graduate Education and Research to aid in doing so. This money was used to defray the ex- penses of having made a series of accurate analyses of certain soils and of the ash of several plants growing on them, the analytical work being carried out in a highly satisfactory manner by Mr. Horace J. Hallowell, a consulting chemist of Philadelphia. The plants were selected for study so as to be representative of important classes, as follows: One fern, one conifer, one monocot, one primitive dicot, and two specialized dicots. In each case, one sample was collected from a serpentine-barren, and another of morphologically similar material from either Piedmont or Coastal Plain woods. My thanks are due to Dr. Francis W. Pennell, who has made a special study of the plants of the serpentine-barrens, for aid in locating several of those here investigated. The species of plants which grow on the serpentine-barrens have migrated there from other geological formations in the same general region, so the first subject to which attention was directed was the com- position of the soil in various localities of these plants. The serpentine- barren soil analyzed consisted of material which adhered to the roots of one of the most characteristic species of such areas, namely Moss Phlox (Phlox suhulata L.) collected 1* miles northeast of Unionville, Chester County, Pennsylvania. To represent the soil of Piedmont woods, a sam- ple was similarly obtained from the roots of Spike Gayfeather (Liatris spicata (L.) Willd.) f mile south of Crozierville, Delaware County, Pennsylvania. The underlying rock here is gabbro, but the plant proved to be morphologically indistinguishable from that growing on serpentine a few miles away. Coastal-plain woods soil was shaken from the roots of a clump of Moss Phlox identical in aspect to that on the serpentine at Chews Landing station, Camden County, New Jersey. These soil samples were sifted through a screen with meshes 1 mm. in diameter to remove coarse rock fragments and plant debris, and the nitrogen and acid-soluble mineral elements were determined by the official methods of the Association of Official Agricultural Chemists, the results being presented in Table 1. Subsequent tables give the data as to the composition of the ash of the several plants studied, manganese and chlorine having been determined by official methods, the remaining elements by those recommended in Washington's Chemical Analysis of Rocks. 1; 34 Acid-soluble PENNSYLVANIA ACADEMY OF SCIENCE TABLE 1 CONSTITUENTS OF SOILS SUPPORTING THE PLANTS INVESTIGATED Serpen- tine- barren Pied- mont woods Coastal- plain woods Serpen- tine- barren Pied- Coastal mont plain woods woods KoO 0.18 0.09 0.14 Na.O 0.03 0.12 0.05 CaO 0.43 0.20 0.24 MgO 17.03 0.33 0.26 Fe,03 ... 8.40 3.01 2.26 AlA ... 1.32 4.50 1.04 PA 0.32 0.22 0.17 SiO, N, total N, as NHs N, as NO3 pH (colorim.) act. acidity 0.08 0.12 0.07 0.47 0.30 0.27 0.06 0.02 0.04 none none none 6.0 5.5 5.0 10 30 + 100 Formed as they have been under essentially identical el^^f « ;°^<^^- tion! the acid-soluble portions of the three soils are -t markedly ^^ ferent in character, except that particles of serpentine distributed through the first yikd relatively large amounts of magnesia and iron "^^'l'- TABLE 2 COMPOSITION OF THE AsH OF BRAKE (PteHdium latiusculnm (DESV.) MAXON) Constit- uents Coastal-plain woods Per cent, of ash Per cent, of plant Serpentine-barren KoO .. isra.o CaO MgO MnO FeoOj AI2O3 P.O5 .. SO, . 01 SiOo . 0 = 01 3.38 1.57 10.02 6.76 0.40 1.45 2.40 2.32 2.47 0.06 69.36 -0.01 0.19 0.09 0.57 0.385 0.02 0.08 0.135 0.13 0.14 0.005 3.955 Per cent, of ash 10.40 2.15 7.98 14.45 0.35 0.66 1.05 2.47 1.68 1.84 57.86 -0.40 Per cent, of plant Change Amount Sum 100.18 5.70 100.49 0.685 0.14 0.525 0.95 0.02 0.04 0.07 0.16 0.11 0.12 3.805 -0.025 -fO.495 -i-0.05 -0.045 +0.565 Per cent. +260 + 55 - 8 +149 -0.04 -0.065 +0.03 -0.03 +0.115 -0.15 - 50 - 48 + 23 - 21 6.60 +0.925 - 4 + 16 Locali- ties West of Albion, Camden Co., N. J. South of Lima, Dela- ware Co., Pa. Notable features : Eatio, basic to acidic oxides, 1:3; ^^l^]'l^^^^^ high ; result of migration into serpentine-barren, marked increase total ash, potash, and magnesia. PENNSYLVANIA ACADEMY OF SCIENCE TABLE 3— Composition op the Ash of Pitch Pine (Pinus rigida Miller) 35 Constit- uents K2O .... NasO . CaO .. MgO . MnO Fe^Oa AI2O3 P2O5 SO3 CI SiOa . 0 = C1 Coastal-plain woods Serpentine-barren Per cent, of ash Sum 27.90 5.66 17.98 6.01 0.80 0.96 10.97 12.79 6.68 0.67 8.96 -0.14 Per cent, of plant 0.76 0.155 0.49 0.165 0.02 0.025 0.30 0.35 0.18 0.02 0.24 -0.005 Per cent, of ash 99.24 2.70 19.02 1.34 19.05 16.64 0.60 1.27 4.16 10.74 10.42 1.44 15.86 -0.29 Per cent, of plant 100.75 0.39 0.025 0.39 0.34 0.01 0.025 0.085 0.22 0.21 0.03 0.32 -0.005 Change Amount Per cent. -0.37 -0.13 -0.10 +0.175 -0.01 2.04 -0.215 -0.13 +0.03 +0.01 +0.08 -0.66 - 49 - 84 - 20 +106 - 50 - 72 - 37 + 17 + 50 + 33 - 24 Locali- ties Atsion, Burlington Co., N. J. Nottingham, Chester Co., Pa. Notable features: Ratio, basic to acidic oxides, 3:1; potash high, silica low ; result of migration into serpentine-barren, marked decrease in total ash and in potash, but increase in magnesia. TABLE 4-CoMPOSiT.ON op the Ash of Catbrier (Smilax rotundifoUa L.) Constit- uents Coastal-plain woods K2O ... Na^O CaO ... MgO MnO FeoOa AI2O3 P,0» SO, ... CI SiOo 0 = 01 Per cent, of ash Sum 29.69 2.22 31.50 4.00 0.82 1.59 2.01 7.85 14.61 0.66 5.64 -0.14 Per cent, of plant 100.45 0.615 0.045 0.65 0.08 0.015 0.035 0.04 0.16 0.30 0.015 0.115 Serpentine-barren Per cent, of ash 2.07 39.06 4.35 14.68 17.01 0.63 0.87 0.40 8.94 11.01 1.54 1.91 -0.30 Per cent, of plant 100.10 0.55 0.06 0.205 0.24 0.01 0.01 0.005 0.125 0.155 0.02 0.025 -0.005 Change Amount Per cent. 1.40 -0.065 +0.015 -0.445 +0.16 -0.005 -0.025 -0.035 -0.035 -0.145 +0.005 -0.09 -0.67 - 16 + 33 - 68 +200 - 33 - 71 - 88 - 22 - 48 + 33 - 78 - 32 Locali- ties West of Albion, Williamson School, Camden Co., N. J. Delaware Co., Pa. Notable features: Ratio of basic to acidic oxides, 3:1;^^^^^^^^^ silica low; result of migration into serpentme-barren^ma^^^^^^^ decrease in total a^h and lime, but a 200 per cent, increase m magnesia. 36 PENNSYLVANIA ACADEMY OF SCIENCE TABLE 5— Composition of the Ash or Blackjack Oak (Quercus manlandica Muench) Constit- uents K2O NaoO . CaO MgO . MnO . Fe^Oa . AI0O3 p^a SO3 CI SiOo ... o = a Coastal-plain woods Per cent, of ash 22.52 5.12 28.40 6.49 0.97 2.83 5.88 8.13 8.11 0.39 n.76 -0.08 Per cent, of plant Sum 100.52 0.605 0.135 0.76 0.175 0.025 0.075 0.16 0.22 0.22 0.01 0.315 Serpentine-barren Per cent, of ash Per cent, of plant 2.70 16.65 1.37 31.24 18.84 1.08 1.00 1.92 8.32 8.79 0.18 11.41 -0.04 100.76 0.365 0.03 0.685 0.41 0.025 0.02 0.04 0.18 0.19 0.005 0.25 Change Amount 2.20 -0.24 -0.105 -0.075 +0.235 -0.055 -0.12 -0.04 -0.03 -0.005 -0.065 Per cent. - 37 - 78 - 10 +134 - 73 - 75 - 19 - 14 - 50 - 21 -0.50 - 19 Locali- ties West of Albion, Camden Co., N. J. Williamson School, Delaware Co., Pa. Notable features : Ratio, basic to acidic oxides, 3:1; potash high, silica low ; result of migration into serpentine-barren, decrease m total ash and potash, but considerable increase in magnesia. TABLE 6— Composition of the Ash of Moss Phlox (Phlox sul)ulata L.) Constit- uents K2O .... Na20 . CaO .... MgO . MnO . Fe^Os AI2O3 P2O5 SO3 CI SiOj .. 0 = C1 Coastal-plain woods Per cent, of ash Sum 14.21 5.27 22.66 3.00 0.17 2.70 6.73 4.59 3.29 3.50 34.06 -0.70 Per cent, of plant 0.72 0.27 1.15 0.15 0.01 0.14 0.34 0.23 0.17 0.18 1.73 -0.03 Serpentine-barren Change Per cent, of ash 99.42 5.06 7.50 4.48 17.52 7.71 0.07 1.07 5.18 2.84 1.32 1.86 51.20 -0.40 Per cent, of plant 100.35 0.465 0.275 1.085 0.475 0.005 0.065 0.32 0.175 0.08 0.115 3.16 -0.02 Amount Per cent 6.20 -0.255 +0.005 -0.065 +0.325 -0.005 -0.075 -0.02 -0.055 -0.09 -0.065 +1.43 +1.125 - 35 + 2 - 6 +217 - 50 - 54 - 6 - 24 - 53 - 36 + 83 + 22 Locali- ties Chews Landing, Camden Co., N. J. NE. of Unionville, Chester Co., Pa. Notable features : Eatio, basic to acidic oxides, about 1:1; result of migration into serpentine-barren, marked increase in total a^h, magnesia and silica, but a decrease in potash. PENNSYLVANIA ACADEMY OF SCIENCE 37 TABLE 7 — Composition of the Ash of Spike Gayfeather (Liatris spicata (L.) WiLLD.) •>! J • J_ Piedmont woods Serpentine-barren Change Constit- uents Per cent, of ash Per cent, of plant Per cent, of ash Per cent, o'f plant Amount Per cent. K2O 16.02 0.81 17.60 5.46 0.35 0.38 1.03 2.22 4.25 3.07 49.64 -0.67 1.355 0.07 1.485 0.46 0.03 0.03 0.085 0.19 0.36 0.26 4.19 -0.055 9.11 1.30 6.80 19.62 0.31 0.84 0.12 1.15 4.04 0.61 56.80 -0.13 0.735 0.105 0.545 1.58 0.025 0.07 0.01 0.09 0.325 0.05 4.565 -0.01 -0.62 +0.035 -0.94 +1.12 -0.005 +0.04 -0.075 -0.10 -0.035 -0.21 +0.375 - 46 Na^O CaO MgO MnO Fe20, AI2O, + 50 - 63 +265 - 17 +133 - 88 p,0, - 53 ■•- 2^-'5 SO3 - 10 CI SiOj 0 = CI - 81 + 9 Sum 100.16 8.46 100.57 8.09 -0.37 - 4 Locali- ties S. of Crozierville, Delaware Co., Pa. Williamson School, Delaware Co., Pa. Notable features : Ratio of basic to acidic oxides, 2:3; result of migration into serpentine-barren, great increase in magnesia, and mod- erate decrease in lime and potash. TABLE 8-Ash-composition of Hairy Cerastium (Cera^tium arvensevillosum rM^HL.-) HOLL. & Brit.) on Two Different Serpentine-barrens Constituents Staten Island' Unionville, Pa. Percentage of ash Percentage of plant 29.63 1.79 4.32 0.26 3.59 0.22 20.72 1.25 0.10 0.005 2.30 0.14 5.98 0.36 9.19 0.555 3.50 0.21 7.77 0.47 13.82 0.835 -1.69 -0.10 Sum Ratio, B : A 99.23 2: 1. 6.00 3 Bull. Torrey Bot. Club, 14: 49, 1887. 38 PENNSYLVANIA ACADEMY OF SCIENCE In this case the plant is endemic on serpentine, so no comparison with occurrences on another type of soil can be made, but analyses of it from two serpentine-barrens 100 miles apart show that while the amount of some constituents varies from one locality to another, the magnesia con- tent remains essentially constant at around 20 per cent. 8ummary.-Ash analyses have been made of six widely different plante growing in serpentine-barrens and in nearby Piedmont or Coastal- plain woods In all cases there was a marked increase m magnesium oxide content when the plants invaded the highly magnesian soil of the barrens accompanied by changes in other constituents showing no regu- larity. The ash of an endemic Cerastium from two different barrens showed uniformity only in high magnesia content. Plants in which the ash-composi- tion has been studied Brake {Pteridium latiusculum) Pitch Pine (Pinus rigida) Catbrier {Smilax rotundifolm) Blackjack Oak {Quercus marilan- dica) Hairy Cerastium {C. arvense vil- losum). Moss Phlox (Phlox subulatd) Spike Gayfeather (Liatris spicata) Ratio, bases to acids 1: 3 3:1 3: 1 3:1 2:1 1:1 2: 3 Results of migration into serpen- tine-barren Increase in MgO, % 149 106 200 134 217 265 Changes in other constituents Total ash and potash increased. Total ash and potash decreased. Total ash and lime decreased. Total ash and potash decreased. Total ash and silica increased. Potash and lime de- creased. Reprinted from American Fern Journal, Vol. 22, No. 3, July-September, 1932. Range-Extensions and other Observations, 1931-32 Edgar T. Wherry^ Pellaea Bridgesii in Oregon. — On June 30, 1931, Dr. Francis W. Pennell and I drove from the largely aban- doned village of Cornucopia, Baker County, Oregon, five miles westward up an exceedingly steep road to the Union Mine, to collect plants on the mountain there. On ledges of granite rock along a trail leading up to some old workings, we found several colonies of a fern which was obviously a Pellaea, but differed from any I had previously seen in having the soral band well in from the margin. This was subsequently identified as P. Iridgesii Hooker. Specimens distributed by Cusick many years before may have come from the same general region, but they lacked definite locality data; and in Abrams' ' ' Illustrated Flora of the Pacific Coast, ' ' Maxon gives the range of the species as ' ' California, from Nevada County southward to Mineral King, Tulare County; also in Boise National Forest, Idaho. ' ' The newly recorded find therefore represents a distinct extension of the hereto- fore recognized range. Cheilanthes gracillima as an Example of Non- CiRCiNATE Vernation. — ^My first acquaintance with this species in its native haunts occurred at the locality men- tioned in the preceding note. The new fronds for the season were just beginning to develop, and proved to show the same type of non-circinate vernation already reported for C. tomentosa and several other f erns.^ 1 Contribution from the Botanical Laboratory of the University of Pennsylvania. 2 Amer. Fern Journ. 16: 107, 109. 1926; 18: 31. 1928. Observations, 1931-32 81 A Visit to a Station for the Appalachian Filmy- Fern. — For some years I have desired to see Tricho- manes hoschianum in its native haunts, but the roads leading to reported localities of it would always prove to be impassable at the times of my visits to the region. In July, 1932, Mr. J. E. Benedict and I were driving through northern Alabama during a period of drought, and decided to hunt up a station located by Mr. E. W. Graves along the Sipsey River. We found the Jasper topographic sheet to be quite useless, as the culture has completely changed since the time it was surveyed, so some data as to the route followed may be added here for the benefit of others who may wish to visit the place. We went north on the paved highway (No. 5) from Jasper, Walker County, about 5 miles to the outskirts of the village of Manchester, and there turned north- eastward on an improved dirt road, going 6 miles to a road fork. Here the left-hand road was taken, and after 3 miles of rather rough travelling, we reached Duncan's Bridge over the Sipsey River. Parking on the north side, we walked east along a trail which followed the talus-piles at the base of the cliffs. About a quarter of a mile in from the bridge, the long-sought filmy-fern . was found on the face of a deeply inset, moisture-laden sandstone stratum. Its soil-reaction proved to be low mediacid, active acidity 100. Referring the locality to the nearest settlement in the same county, it may be designated as: 3 miles south of Mellville, Winston County, Alabama. The Alabama Colony of Scott's Spleenwort.— Three years ago, as recorded by myself and Mr. Harry W. Trudell,^ a visit was made to the station for Asplenium ehenoides near Havana, Alabama. At that 3AMEE. Fern Joubn. 20: 30. 1930. Observations, 1931-32 81 i A Visit to a Station for the Appalachian Filmy- Fern. — For some years I have desired to see Tricho- manes hoschiamcm in its native haunts, but the roads leading to reported localities of it would always prove to be impassable at the times of my visits to the region. In July, 1932, Mr. J. E. Benedict and I were driving through northern Alabama during a period of drought, and decided to hunt up a station located by Mr. E. W. Graves along the Sipsey River. We found the Jasper topographic sheet to be quite useless, as the culture has completely changed since the time it was surveyed, so some data as to the route followed may be added here for the benefit of others who may wish to visit the place. We went north on the paved highway (No. 5) from Jasper, Walker County, about 5 miles to the outskirts of the village of Manchester, and there turned north- eastward on an improved dirt road, going 6 miles to a road fork. Here the left-hand road was taken, and after 3 miles of rather rough travelling, we reached Duncan's Bridge over the Sipsey River. Parking on the north side, we walked east along a trail which followed the talus-piles at the base of the cliffs. About a quarter of a mile in from the bridge, the long-sought filmy-fern was found on the face of a deeply inset, moisture-laden sandstone stratum. Its soil-reaction proved to be low mediacid, active acidity 100. Referring the locality to the nearest settlement in the same county, it may be designated as: 3 miles south of Mellville, Winston County, Alabama. The Alabama Colony of Scott's Spleenwort.— Three years ago, as recorded by myself and Mr. Harry W. Trudell,^ a visit was made to the station for Asplenium ehenoides near Havana, Alabama. At that 3 Amer. Fern Journ. 20: 30. 1930. 1 Observations, 1931-32 83 H < ip-i P O •J < o o time only 25 adult plants were seen, although there were numerous young ones. On July 13, 1932, I revisited the locality, this time in company with Mr. J. E. Benedict, Jr., and was glad to find that many of the young plants formerly observed have reached maturity, and at least 85 adult ones were counted in the same area as was pre- viously examined. As before, there was no Walking- fern on those rock ledges, but subsequently Mr. William A. Knight, of Biltmore Forest, N. C, has informed me that there is a colony on the opposite side of the same stream where this fern does grow with its offspring. Dr. Kestner's Results with Hybrid Aspleniums. — The experiments in growing ferns from spores being carried on by Dr. Paul Kestner of Lausanne, Switzer- land, have already been referred to in this Journal.^ Spores from several of the presumably hybrid Asple- niums from our eastern states have been sent to him for trial, and he has communicated to me the results which form the basis for the following notes. Of all the spores of Aspleniitm ehenoides which he has received from various parts of this country, only those from the above-mentioned Alabama colony have proved to be viable. Evidently in most of its occurrences this fern represents a hybrid, which has not attained fertility; but at this single locality one cross, at some past time, chanced to produce viable spores, and its descendants have remained fertile, so that it has become a true species. The spores of Asiylenium stotleri have germinated well, and a pressed plant only two years old sent me by Dr. Kestner reproduced in a most striking way all the features shown by those in the original colony. It therefore likewise represents a hybrid (presumably of A. pinnatifidum X A. platyneuron) which chanced to 4 Amer. Fern Journ. 19: 60. 1929; 21: 29. 1931. Observations, 1931-32 83 time only 25 adult plants were seen, although there were numerous young ones. On July 13, 1932, 1 revisited the locality, this time in company with Mr. J. E. Benedict, Jr., and was glad to find that many of the young plants formerly observed have reached maturity, and at least 85 adult ones were counted in the same area as was pre- viously examined. As before, there was no Walking- fern on those rock ledges, but subsequently Mr. William A. Knight, of Biltmore Forest, N. C, has informed me that there is a colony on the opposite side of the same stream where this fern does grow with its offspring. Dr. Kestner's Kesults v^tith Hybrid Aspleniums. — The experiments in growing ferns from spores being carried on by Dr. Paul Kestner of Lausanne, Switzer- land, have already been referred to in this Journal.^ Spores from several of the presumably hybrid Asple- niums from our eastern states have been sent to him for trial, and he has communicated to me the results which form the basis for the following notes. Of all the spores of Asplenium ehenoides which he has received from various parts of this country, only those from the above-mentioned Alabama colony have proved to be viable. Evidently in most of its occurrences this fern represents a hybrid, which has not attained fertility; but at this single locality one cross, at some past time, chanced to produce viable spores, and its descendants have remained fertile, so that it has become a true species. The spores of Asplenium stotleri have germinated well, and a pressed plant only two years old sent me by Dr. Kestner reproduced in a most striking way all the features shown by those in the original colony. It therefore likewise represents a hybrid (presumably of A. pinnatifidum X A. platyneuron) which chanced to 4 Amer. Fern Journ. 19: 60. 1929; 21: 29. 1931. 11 84 American Fern Journal Observations, 1931-32 85 attain fertility, and, successfully reproducing itself, has become a species. Growing as it does only on a single small cliff (in Jefferson County, West Virginia), it is to be regarded as an example of an endemic in the re- stricted sense, ie., a plant occupying a small area on the earth 's surface because it is of too recent origin to have become dispersed more widely. Perhaps it will become extinct before being able to spread, for as the result of a series of excessively dry seasons the original colony has dwindled considerably. But at least it will be preserved in cultivation in the collection assembled by Dr. Kestner. On the other hand, the spores of all specimens of A, trudelli sent thus far (from Georgia and Pennsylvania) have proved to be imperfect and non-viable, clearly in- dicating that the particular plants from which the spores came represented first-generation hybrids (of A. mon- tanum x A. pinnatifidum ) . Search for a colony in which fertility has been attained will, however, be continued. At this point the hope may perhaps be expressed that some members of the American Fern Society in our own country will take up the growing of these rarer ferns from spores. I should be only too glad to cooperate by furnishing spore material and data as to soil acidity preference of each species or hybrid. Further Occurences of the Rocky Mountain Cliff- Fern IN THE East.— The eastern relative of Woodsia scopuUna has been recorded thus f ar^ from three stations in Virginia, one in "West Virginia, and two in North Carolina. Two new finds of it may here be noted. First, Mr. Arthur N. Leeds of the Academy of Natural Sciences of Philadelphia discovered on August 3, 1931, an addi- tional one in the first-named state, on the west side of the Cowpasture River north of Longdale, Alleghany County. Then, in July, 1932, I had the pleasure of sAmer. Fern Journ. 19: 101. 1929. visiting the fern garden of Mr. William A. Knight, at Biltmore Forest, North Carolina, and found that he had growing there several clumps of an unidentified rock- fern from the cliffs of the Nolichucky River in Unicoi County, Tennessee. On examination it proved to repre- sent this as yet undescribed Woodsia, thus extending its known range into an additional eastern state. The Limestone Adderstongue in Northern Virginia. —Mr. Palmer's recent article on Ophioglossum engel- manni^ gives a good account of the occurrence of this fern, but his locality list, being based on only two herbaria, does not represent a complete statement of its distribution. Two states in which it is known are not mentioned : its discovery in Ohio was announced by Miss Braun several years ago," and in the newly issued *' Ferns of Florida, ' ' Dr. Small states that ' ' Recent exploration has discovered this adders-tongue in abundance in the western part of northern Florida. In Virginia only two occurrences near Natural Bridge are cited by Mr. Palmer ; the county name of these, given as * * Breckenridge, " should read Rockbridge. It has also long been known near Staunton, Augusta County, and attention may here be called to its occurrence still further north. In August, 1925, as reported in this Journal^ I found it in association with a new species of Opuniia near Luray, Page County. In June of the present year, while accompanying Miss Lena Artz of AVoodstock, Shenandoah County, in a search for native plants in that vicinity, another extensive colony of this cactus was observed on similar limestone ledges 2\ miles northeast of the town, and I suggested that a search for the fern be made there. A few days later she was suc- 6 Amer. Fern Jouen. 22: 43. 1932. 7 Amer. Fern Journ. 17: 138. 1927. 8 Amer. Fern Journ. 16 : 2. 1926. 86 American Fern Journal cessful in finding it, and reports that still later, follow- ing a long-awaited rain, it came up in abundance. This extends its previously known range 15 miles northward, to latitude 38° 55'. The Climbing Fern in North Carolina.— In the list of ferns of this state recently published by Mr. Blom- quist,» Lygodium palmatum is indicated as occurring only 'in the western part, that is, in the Blue Ridge physiographic province. Several years ago I was guided by Dr P. 0. Schallert, of Winston-Salem, to the Cas- cades near Danbury, in the Piedmont province, and in wet woods near by found this fern in the greatest profu- sion In March, 1932, Mr. Rogers McVaugh, a graduate student in the Department of Botany of the University of Pennsylvania, found the same species two miles west of Warsaw, in Duplin County, on the Coastal Plain. So it is evidently widely distributed over the state. Philadelphia, Pa. 9 Amer. Fern Journ. 21 : 86. 1931. r »— Wll«»^mi«— «H»— l»»«»^ltM^^IIII^^IIM— HH ■■«^»a«y I Some Forgotten Records of Hybridization and Sex in Plants Conway Zirkle Reprinted without change of paging j from the Journal of Heredity (Organ of I the American Genetic Association), Wash- I ington, D. C, Vol. XXIII, No. 11. Novem- ber, 1932. I 1 A » II FIRST OBSERVED HYBRIDIZATION IN PLANTS His Witch-hunting activities have led posterity to ^-^^|^^> ^^%^,"^^^^^^ Mather "contributed my two mites to the way wherem vegetation is earn i o"' ^" contributio^i is the earliest recorded observation of natural hybridization in plants. SOME FORGOTTEN RECORDS OF HYBRIDIZATION AND SEX IN PLANTS 1716-1739 Conway Zirkle Department of Botany, University of Pennsylvania THE first half of the eighteenth century was a time of exceptional botanical activity in the English colonies along the Atlantic Coast of North America. A new country filled with strange and interesting plants had just been opened for exploration and European botanists were anxious for specimens. Medicinal plants were in great demand and any educated colon- ists who sent seed or herbaria sheets to England or the Continent could be as- sured of an interesting and profitable correspondence with the plant im- porters. The colonists who collected, systema- tized and recorded the distribution of the new plants seem to have been ex- ceptionally able. They kept in close touch with the development of botany in Europe, visited and wrote letters to each other and exchanged ideas and specimens. They imported microscopes, investigated plant anatomy, especially the anatomy of the flower, and devised a number of physiological experiments. The most prominent men in the colon- ies were keenly interested in this local scientific development and some even joined in the experimentation. The European botanists were cer- tainly informed of the work of the Americans, although the very real con- tributions made by the latter, in fields other than taxonomy, have been, with a single exception, completely forgotten. Most biologists have been so im- pressed by the overwhelming adequacy of Sachs' History of Botany (1530- 1860) that any important botanical con- tribution which he failed to record is apt to remain unknown. Moreover, so greatly is Sachs' judgment admired and his fairness recognized (except perhaps in his treatment of Linnaeus), that he has become the principal arbiter of priority claims even in such a con- fused subject as that of the discovery of sex in plants. Sachs' evaluation of the contributions of the different work- ers in this field is still generally ac- cepted. In order to prove that plants repro- duce sexually it is necessary to show, first, that viable seeds can not be pro- duced without the cooperation of some element (pollen) which might be inter- preted as male, and, second, that both this hypothetical male element and the egg bear factors which influence the progeny. Camerarius (1694) is cited by Sachs as the first investigator to prove experimentally that pollen is nec- essary for seed development, while Koelreuter (1761) is credited with hav- ing made the first systematic study of plant hybridization, proving incidentally that both parents contribute to the off- spring. Sachs rightly emphasized the importance of Koelreuter's work for, as far as he knew, there were no reliable records of plant hybrids before 1761. In fact he mentions only two instances of species crossing before Koelreuter's experiments and both were considered somewhat dubious. He said of the first, "Bradley is our authority for the statement that a gardener in London had obtained a hybrid between Dianthus caryophylhis and Dianthus harhatns by artificial means as early as 1719" (Sachs, p. 406). The second instance is re- corded just as briefly, "Soon after allu- sion is made (by Linnaeus, 1735) to the artifices used by gardeners to obtam hybrid tulips and cabbages, but the mat- ter is treated rather as agreeable tri- fling" (Sachs, p. 400). Haartman on 433 434 The Journal of Heredity Zirkle : Early Records of Plant Hybrids 435 I ii III III !l! purely taxonomic grounds had interpret- ed some of his finds as natural hybrids. Cotton Mather Forty-five years before Koelreuter crossed different species of Nicotiana, however, an American, apparently un- known to Sachs, had observed three im- portant phenomena: (1) ^^^^ pollina- tion; (2) hybridization (variety and perhaps species crosses) ; and (3) the resemblance of the offspring to the male parent. Cotton Mather is better known today as a witch-hunter than as a member of the Royal Society. While he was un- doubtedly very credulous and believed on very scanty evidence much that was later shown to be false, he at least kept in touch with the scientific progress of his time and contributed what he could to the advancement of knowledge. We know that he was informed of the new hypothesis, that the flowering plants re- produced sexually, and he evidently ac- cepted the views advanced by Nehemiah Grew, as is shown by the following pas- sage (Mather, 1721) : The Stamina^ with their A f ices; and the Stylus (called the Attire bv Dr. Grezv) which is found a sort of Male Sferniy to im- pregnate and fructify the Seed ! The observations of Cotton Mather, which should take their proper place in the history of botany, were described by him in a letter to James Petiver, F. R. S., dated September 24, 1716. This letter, which is now in the files of the British Museum (Sloane Ms. 4065, fol. 255), has probably never been pub- lished, but Kittredge (1916) has sum- marized its contents. In it Mather de- scribed hybridization in Zea and Cucur- bita. He repeated the description in his book, Religio Philosophica; or The Christian Philosopher, London, 1721. The following passage is taken from Essay XXVI, Of the VEGETABLES. That I may a litde contribute my two Mites to the illustration of the way wherein Vegetation is carried on, I will here com- municate a couple of Experiments lately made in my Neighbourhood. My Neighbour planted a Row of Hills in his Field with our Indian Corny but such a Grain as was coloured red and Mue; the rest of the Field he planted with Corn of the most usual Colour, which is yellow. To the most Windzvard-side this Row infected jour of the next neighbouring Rows, and part of the fifth, and some of the sixth, to render them colour'd like what grew on itself. But on the Leeward-side no less than seven or eight Rows were so colour'd, and some smaller impressions were made on those that were yet further distant. The same Neighbour having his Garden often robb'd of the Squashes growing in it, planted some Gourds among them, which are to appearance very like them, and which he distinguished by certain adjacent marks, that he might not himself be imposed upon; by this means the Thieves 'tis true found a very bitter Sauce^ but then all the Squashes were so infected and embittered, that he was not himself able to eat what the Thieves had left of them. As the bitterness of the squashes fol- lowing their mesalliance with the gourds appears a generation too soon, it is regretable that a more careful rec- ord was not kept. For until we can trace Fairchild's hybridization of Dian- thus to some period before 1717, Math- er's account is the earliest we have of possible species crossing in plants.* The spontaneous crossing of different varieties of Zea Mays was certain to at- tract attention sooner or later. Doubt- less many more records will be found when the documents of this period have been examined adequately. It is not strange that Cotton Mather's account of cross pollination was overlooked as the biologists of his time were suspicius of the New England "Saints" and did not search their theological works for scientific data. However, it is remark- able that other records of this hybridi- zation have not received more attention. Thomas Fairchild ♦The gourds were Cucitrbita Pepo var. ovifera, the squashes were probably either C. Pepo var. condensa or C. maxima; if the latter, Mather reported a real species cross. The first artificial hybrid, of which we have a record, was made by Mr. Thomas Fairchild, of Hoxton, when he crossed Dianthus caryophyllus with Dianthus barbatus. Although Fairchild' s contemporaries gave him full credit for his achievement, Sachs minimized it and the fashion was set which has lasted unchallenged to the present. ^ Sachs^^ did not even mention Fairchild's name, but merely referred to him as a "gardener in London." Focke^^ cited Fairchild's successful experiment (pp. 55, 430) but added, "This success in artificial fertili- zation was neither used for the ad- vancement of science nor does it seem to have given gardeners any stimulus for further research." Pfeffer^^ went even further by stating (p. 265), "Die wohl friiher (1719) ausgeffuhrt Kreuzung zweier Nelken durch Fairchild war ein rein gartnerischer Versuch, der keine wissenschaftigen Bedeutung erlan^te." Strangely enough even the recent Eng- lish and American historians of botany (Green,i« Roberts^^) have failed to combat ^his completely unfair estimate of Fairchild's contribution. Unfortunately no one has found a really satisfactory description of the Dianthus hybridization. Botanists since Sachs, have seemingly known of this work through a single reference in Bradley's New Improvements in Gar- dening, etc., which is uniformly dated 1719 This reference is quoted in part by both Greeni« ^^d Roberts.^o While this record is better than any other yet found, it is incomplete and should be supplemented by other citations. Ihe following quotation is taken ^^i^om the first edition of Bradley's work which appeared in 1717. Fairchild s experi- ments were, of course, performed ear- Her. From Bradley (1717), part I, pp. 23, 24 : I believe I need not explain how the Male Dust of Plants may be convey'd by the Air from one to another, by which the Gen- eration and Production of new Plants is brought about .... Moreover a curious Person may by this knowledge produce such rare Kinds of Plants as have not yet been heard of, by making choice of two Plants for his Purpose, as near alike in their Parts, but chiefly in their Flowers or Seed-Vessels; for example the Carnation and Sweet Wil- liam are in some respects alike; the Farina of one will impregnate the other, and the Seed so enlivened will produce a Plant dif- fering from either, as may now be seen in the Garden of Mr. Thomas Fairchild^ of Hoxton, a Plant neither Sweet William nor Carnation, but resembling both equally; which was raised from the Seed of a Car- nation that had been impregnated by the Farina of the Sweet William. These Coup- lings are not unlike that of the Mare with the Ass which produces the Mule, and in regard to Generation, are also the same with Mules, not being able to multiply their Sfe- cies, no more than other Monsters generated in the same manner. Bradlev suggests a practical applica- tion of Fairchild's discovery (Part II, pp. 84, 85): ... 1 have endeavoured to explain how the Dust of one Flower will impregnate and enliven the Seeds of another, and that from that accidental Coupling the Seeds are so chang'd as to produce Plants with Blossoms varying from those of the Mother plant. 1 have likewise shewn why double Flowers sel- dom bear Seed, which I conjecture is be- cause the Male Parts in them are either not perfect, or else are confin'd from Action by the Multiplicity of the Petals. This Con- sideration leads me to advise the Curious Florists to plant of every good sort of his double Carnations in Beds on a Line in the Middle, and on each Side of them to set at least Two Rows of single ones of choice Colours, and among them some Plants of Sweet William, and of the^ China ox Indian Pinks, which have such Varieties of odd Col- ours in them, as I shall mention hereafter The China Pinks and the Sweet Wtlhams bearing single Flowers, as well as the single Carnations, may have Opportunities of com- municating their Farina into the CeUs of the double ones, and set their Seeds, which if they do, we shall not only gather a larger Quantity than we could otherwise expect, but likewise be assur'd of great Varieties from them. 436 The Journal of Heredity Zirkle: Early Records o£ Plant Hybrids 437 The following references supplement Bradley. Miller (1731) under the heading CARYOPHYLLUS, has: 7. CARYOPHYLLUS; barbatus, hor- tensis, angustifolius, flore fleno rosto. The Double Rose-colour'd Sweet John or Fair- child's Mule .... these continue flowering for a long time, and are extremely beau- tiful, especially the Mule, which produces two full Blooms of Flowers, one in M^y, and the other in Jul*^ .... This passage remained in The Gar- dener s Dictionary until 1754. Perhaps the best description of the hybrid is to be found in a letter of Peter CoUinson to John Bartram dated July 22, 1740 (Darlington, 1849). An instance we have in our gardens, raised by the late Thomas Fairchild, who had a plant from seed, that was compounded of the Carnation and Sweet William. It has the leaves of the first, and its flowers double like the Carnation— the size of a Pink— but in clusters like the Sweet William. It is named a Mule—ptr analogy to the mule produced from the Horse and Ass. If the following record of the Miile lacks exactitude its source justifies its ■ inclusion. Erasmus Darwin was not only a poet, a physician and a grand- father, but also a well informed botan- ist. He knew of Fairchild's hybrid and described it in 1781 thus: Botanic Garden^ Part II, Canto IIII CARYO's sweet smile DIANTHUS proud admires. And gazing burns with unallow'd desires; With sighs and sorrows her compassion moves. And wins the damsel to illicit loves. The Monster-ofl"spring heirs the father's pride. Masked in the damask beauties of the bride. When Miller adopted the Linnaean system in 1759 he included the follow- ing under the heading '^Dianthus'^ : 3. The Mule, or Fairchild's Sweet Wil- liam, it hath narrower Leaves than either of the former, and is of that Variety called Sweet John: This was said to have been pro- duced from Seeds of a Carnation, which had been impregnated by the Farina of the Sweet William: The Flowers of this are of a brighter red Colour than either of the former, their Bunches are not quite so large, but the Flowers have an agreeable Odour. When Thomas Martyn revised the dictionary in 1807, the above passage was slightly condensed. The references thus far quoted do not give us a sufficient basis for judg- ing the character of Fairchild's work. Fairchild himself pufblished very little: The City Gardener in 1722 and a short account of some remarkable ex- periments in the Philosophical Transac- tions of 1724. By far the greater part of his investigations were recorded by his contemporaries: Blair (1720), Brad- ley (1717, 1721, 1726), Miller (1731) and Collinson (1740). Space forbids an account of Fairchild's many activi- ties. That he was the outstanding commercial florist of his time is shown by many easily obtainable refer- ences (Blair, 1720; Bradley, 1717, 1726; Pulteney, 1790; Nichols, 1817; Felton, 1830; Britten and Boulger, 1893). Bradley (1726) referred to Fairchild over twenty times and devoted fifteen pages (Vol. II, pp. 458-473) merely to listing the flowers in the gar- den at Hoxton with the times of bloom- ing, including in the list the Mule be- tween the Sweet William and Carnation. It was, however, as an experimental biologist that Fairchild made his chief contributions. At a time when the prestige of the Ancients was still high, when analogy was supposed to furnish proof, and when philosophers still knew enough of the Creator's purposes to arrive at the truth by contemplating the Nature of Things, Fairchild experi- n^^nted— both for himself and for the more dignified philosophers. His rela- tion to his co-workers is well shown by the honest acknowledgment of ^^^^13^ the preface to Botanical Essays (1720: ."Mr. Fairchild's {whom I have often mentioned, and to whom I owe all the practical observations I have advanc d concerning the Vegetation):' Fairchild also performed many of Bradley's ex- periments in addition to the most curi- ous and ingenious ones of his own de- vising (Phil. Trans., Number 384). To label the hybridization of Dian- thus by the leading experimenter of his generation as '*ein rein gartnerischer Versuch, der keine wissenschaftigen Bedeutung erlangte," in order to give Koelreuter a priority he neither needs nor deserves, would be mildly humorous if such mistatements were not repeated indefinitely. Philip Miller Philip Miller was the most widely known botanist of his generation. In his own lifetime his Gardener's Dic- tionary j>assed through eight full and six abridged editions, together with three editions in foreign languages. Yet in spite of this, his contributions toward establishing the theory of sexual repro- duction in plants have been noted only in part and his records of plant hybrid- ization have been ignored. Miller is credited with being the first investigator to describe insect pollina- tion. Sachs^^'^^ erroneously gives the date of this occurrence as 1751, al- though it had been recorded many times much earlier. From Sachs^^ "Von be- sonderem Interesse sind die spateren Versuche Miiller's (Miller's, in the English translation) von 1751, welche Koelreuter aus dem Gartner slexicon (II Theil p. 543) mittheilt, insofern hier zum ersten Mai die Insectenhiilfe bei der Bestaubung beobachtet wurde." Sachs, as usual, used Koelreuter's cita- tions and Koelreuter found the refer- ence in the German edition of the Gar- dener's Dictionary, Nurnberg, 1750-51. Green records Miller's observation by quoting directly from the so-called first edition published in 1731, while Rob- erts quotes from the seventh edition (1759). Miller is also credited with noting that when female spinach plants were raised apart from the male, they pro- duced seed which contained no embryos. However, his other investigations of sexual reproduction in plants (Cucum- bers and Melons) have been left out of the more recent records and his descrip- tion of spontaneous variety crosses in Cabbage is nowhere evident. This is all the more remarkable because he de- scribed his work on Cucumis and Bras- sica in many editions of his dictionary. Miller actually performed his experi- ments and reported his observations much earlier than o\^r current records would indicate. The date assigned by Sachs, 1751, is of course much too late. Green does not state that Miller's inves- tigations were first published in 1731 but merely quotes from the first folio edition of the dictionary. MialF'* quotes from the first octavo edition issued in 1724. Miller's first description of his work, however, did not appear in one of his own books. He sent a full ac- count of his observations on insect pollination and his experiments with spinach to Richard Bradley in a letter dated October 6, 1721. Bradley later published this letter in his Treatise of Husbandry and Gardening, etc., Lon- don, 1726, although an account of Mil- ler's discoveries had already appeared in print. Miller also informed Mr. Patrick Blair of his discoveries in two letters written on October 19th and November 11th, 1721. Blair recognized the im- portance of Miller's results and sent an account of them by letter to Sir Hans Sloane. This letter was published in the Philosophical Transactions, Num- ber 369. Observations ufon the Generation of Plants, in a letter to Sir Hans Sloane, Bart. Pr. Coll. Med. Patrick Blair, M.D., F.R.S. Boston, Dec. 31, 1721. Honoured Sir, It is no small Satisfaction, that what I advanced in my Botanick Esseys is now so fully confirmed by Experiments made by some curious Gardeners, among whom is Mr. Philif Miller, who writes me, November 11, 1721. 1. That in Pursuance of my Advice he separated the Male Plants of the Spinage from the Female; the Consequence was, that the Seeds did swell to the usual Big- ness; but when he sowM it, it did not grow afterwards. He searched Into the Seed, \ i Zirkle : Early Records of Plant Hybrids 439 I' PAUL DUDLEY— JURIST AND NATURALIST The naturalists of Colonial America were many of them ^f^^^^.^J" .^^l^j^\' I^^^Jj Paul Dudley, Attorney General of Massachusetts m 1702, and a f el ow of he Ko> Society was one of the first to discuss the effects of cross polhnation m maize. and found it wanted the Punctutn Vitae, which perhaps might have been the Case with Mr. Geoffrey; but if not, the female Embryones might have been impregnated another Way, as he experimented with twelve Tulips, which he set by themselves about six or seven Yards from any other, and as soon as they blew, he took out the Stamina so very carefully, that he scattered none of the Dust, and about two Days after- wards, he saw Bees working on the Tulips, in a Bed where he did not take out the Stamina^ and when they came out, they were loaded with the Dust on their Bodies and Legs: He saw them fly into the Tulips, where he had taken out the Stamina^ and when they came out, he went and found they had left behind them sufficient to im- pregnate their Flowers, for they were good ripe Seed; which persuades him, that the Farina may be carried from Place to Place by Insects, and when they happen upon a Flower, whose Uterus is capable to be im- pregnated by such a Dust, it may be thus effected. I am of Opinion, this will not suit with Mr. MorlantPs Scheme. For tho' we may suppose the Stamina of every Flower to be loaded with a due Proportion of the Farina^ vet this accidental Conveyance of it to a neighbouring Flower, may be rather less than greater than is necessary: So that, if wanting, then those Embryones^ which had not received its determined Particle into their Bosom, must be defective in Bulk, or barren in growing, but here all were equally fiird. 2. By a Second Letter, October 19, 1721, he informs me, that he bought a Parcel of Savoy Seeds of a Neighbour, which he sowed, and planted out the Plants; but was surprised to see the Production: For he had half of them red Cabbages, and some white Cabbages, and some Savoys with red Ribs, and some neither one Sort nor other, but a Mixture of all Sorts together in one Plant. He went to the Gardiner and told him his Tale, who shew'd him, that he was in the same Condition, but did not know how it should come to pass, for he was sure he took special Care in saving of the Seed. Being askM how and where he planted them for Seed, he shew'd him them under a South- West Hedge, and told him the Manner in which he planted them: First, a Dozen of white Cabbages, then a Dozen of Savoys, and then a Dozen of Red. Then he im- mediately thought how it came to pass, by the Effluvia impregnating the Uterus of one another; and it is very common for our Gardiners to plant white and red Cabbages together for Seed, and they are as often dis- appointed by having a Degeneracy of both Kinds, which they attribute to the Soil, and think that is the Cause: They send to Hol- land for a fresh Supply of Seeds, and say our Soil will not continue that Sort Good. He told them his Opinion, and they laugh at him for it, and will not be turnM out of their Road, although they should have never so many Experiments shew'd them But it is Time to proceed to another Ex- periment of my Correspondent, Mr. Miller, Being persuaded to it by an ingenious Gardiner, he pull'd off all the Male-Flowers of some Melon Plants so soon as they ap- peared; but instead of finding, as his Friend informed him, that these Flowers exhausted the Nourishment from the Fruit; he found that, without these Flowers none of the Melons would grow, for that he was de- prived of the Fruit which he expected. Paul Dudley One early record of hybridization in Zea Mays did receive publicity and was incorporated twice by Philip Miller in the Gardene/s Dictionary. London, 1731, under the headings "GENERA- TION" and "MAYS : Indian Wheat." The source of Miller's information con- cerning the spontaneous crossing of dif- ferent varieties of this New World form was a paper by Paul Dudley, writ- ten in New England and sent to the Royal Society. The communication was entitled "Observations on some of the Plants in New England, with remark- able Instances of the Nature and Power of Vegetation," and was published in the Philosophical Transactions, number 385. Paul Dudley was a member of a dis- tinguished Massachusetts family. In 1702 he l>ecame Attorney General of the Colony and in 1718 a Judge of the Superior Court. He was a Naturalist lit 440 The Journal of Heredity Zirkle: Early Records of Plant Hybrids 441 ■ i :i !i and a Fellow of the Royal Society. The following extract describing variety crosses in Zea Mays is taken from his paper of 1724. It has escaped the at- tention of the historians of botany. The mention of Indian Corn obliges me to take notice of an extra-ordinary Phae- nomenon in the Vegetation of that Gram, viz, the interchanging, or mixing, of Col- ours after the Corn is planted. For your better understanding this Matter, I must ob- serve, that our Indian Corn is of several Colours, as blue, white, red, and yellow; and if they are planted separately, or by them- selves, so that no other Sort be near them, they will keep to their own Colour, i, e.y the blue will produce blue, the white, white, etc. But if in the same Field, you plant the blue Corn in one Row of Hills (as we term them) and the white, or yellow, in the next Row, they will mix and interchange their Colours; that is, some of the ears of Corn, in the blue Corn Rows, shall be white, or yellow; and some again, in the white or yel- low Rows, shall be of a blue Colour. Our Hills of Indian Corn are generally about four Foot assunder, and so continued in a straight Line, or Row of Hills, and so on; and yet this mixing and interchanging of Colours has been observed, when the D'stance be- tween the Rows of Hills, has been several Yards; and a worthy Clergyman, of an Island in this Province {The Reverend Mr. May- hew, of Martha's Vineyard), assures me that the blue Corn has thus communicated, or exchanged, even at the Distance of four or five Rods; and, particularly in one Place, where there was a broad Ditch of Water be- twixt them. Some of our People, but es- pecially the Ah-Origines, have been of the Opinion, that this Commixtion, and Inter- change, was owing to the Roots, and small Fibres reaching to and communicating with one another; but this must certainly be a Mistake, considering the great Distance of the Communication, especially at some Times, and cross a Canal of Water; for the smallest Fibres of the Roots of our Indian Corn, cannot extend above four or five Foot. 1 am therefore humbly of Opinion, that the Stamina, or Principles of this wonderful Copulation, or mixing of Colours, are car- ried thro' the Air by the Wind; and that the Time, or Season of it, is, when the Corn is in the Earing, and while the Milk is in the Grain, for at that Time, the Corn is in a Sort of Estuation, and emits a strong Scent. One Thing, which confirms the Air's being the Medium of this Communication of Colours in the Corn, is an Observation of one of my Neighbours, that a close, high board Fence, between two Fields of Corn that were of a different Colour, entirely prevented any Mixture or Alteration of Col- our, from that they were planted with. The above communication was the basis of all of Miller's accounts of the variety crosses in Zea Mays which ap- peared in the successive editions of The Gardener's Dictionaf y/. I have seen it in the 1st Octavo Edition (1724), 1st Foho edition (1731), 2nd edition (1733), 7th edition (1759) and 9th edition (1807), and also in the 4th and 5th Octavo (Abridged) Editions (1754, 1763). James Logan Another American to recognize the advantages offered by Zea Mays for an investigation of sex in plants was James Logan, a Colonial Governor of Penn- sylvania. He designed several scienti- fic experiments, duly ran the necessary controls, and succeeded in showing that Indian corn would not produce grain if pollen were kept away from the stig- mas. An account of this work was writ- ten in Latin and published in Holland (Experimenta et Maletemata de Plan- tarmn Generatione, Leiden, 1739), and later reprinted in London together with an English translation {Experiments and Considerations on the Generation of Plants, London, 1747). Logan's work was known to the European Botanists of his time and Sachs gave him full credit for his dis- coveries. However, Sachs states in a foot note that he was unacquainted with Logan's Essay and based his account of Logan's contributions on Koelreuter's citations. This essay has not been read- ily available. Harshberger^^ reprinted it in part, unfortunately with modern- ized spelling and punctuation, and Rob- erts^^ has quoted from it extensively. Logan actually performed his experi- ments some years before 1739, for on November 20, 1735, he sent a detailed description of them to Peter Collinson, who submitted the account to the Royal Society. His letter to Collinson was published in the Philosophical Trans- actions, number 440. The following ex- tract is the earliest record I have found of Logan's investigations. Some Experiments concerning the Im- pregnation of the Seeds of Plants, by James Logan, Esq; Communicated in a Letter from him to Mr. Peter Collinson, F. R. S. Philadelphia, Nov. 20, 1735. SIR, As the Notion of a Male Seed, or the Farina Foecundans in Vegetables is now very common, I shall not trouble you with any Observations concerning it, but such as may have some Tendency to what I have to men- tion—And, first, I find from Miller's Dic- tionary, that M. Geoffroy, 2i Name I think of Repute amongst Naturalists, from the Ex- periments he made on Mayze, was of Opin- ion, that Seeds may grow up to their full Size, and appear perfect to the Eye, without being impregnated by the Farina, which possibly, for ought I know may in some Cases be true; for there is no End of Varie- ties in Nature; — but in the Subject he has mentioned I have Reason to believe it's otherwise, and that he applied not all the Care that was requisite in the Management. When I first met with the Notion of this Male Seed, it was in the Winter Tinie, when I could do no more than think of it; but in the Spring I resolved to make some Experiments on the Mayze, or Indian Corn. In each Corner of my Garden, which is forty Foot in Breadth, and near eighty in Length, I planted a Hill of that Corn, and watching the Plants when they grew up to a proper Height, and were pushing out both the Tassels above, and Ears below; from one of those Hills, I cut off the whole Tassels, on the others I carefully open'd the Ends of the Ears, and from some of them I cut or pinch'd off all the silken Filaments; from others I took about half, from others one fourth and three fourths, etc., with some Variety, noting the Heads, and the Quan- tity taken from each: Other Heads again I tied up at their Ends, just before the Silk was putting out, with fine Muslin, but the Fuzziest or most Nappy I could find, to pre- vent the Passage of the Farina; but that would obstruct neither Sun, Air, or Rain. I fastened it also so very loosely, as not to give the least Check to Vegetation. The Consequence of all which was this, that of the five or six Ears on the first Hill, from which I had taken all the Tassels, from whence proceeds the Farina, there was only one that had so much as a single Grain in it, and that in about four hundred and eighty Cells, had but about twenty or twen- ty-one Grains, the Heads, or Ears, as they stood on the Plant, look'd as well to the Eye as any other; they were of their proper Length, the Cores of their full Size, but to the Touch, for want of the Grain, they felt light and yielding. On the Core, when di- vested of the Leaves that cover it, the Beds of Seed were in their Ranges, with only a dry Skin on each. In the Ears of the other Hills, from which I had taken all the Silk, and in those that I had cover'd with Muslin, there was not so much as one mature grown Grain, nor other than as I have mentioned in the first: But in all the others, in which I had left Part, and taken Part of the Silk, there was in each the exact Proportion of full Grains, according to the Quantity or Num- ber of the Filaments I had left on them. And for the few Grains I found on one Head in the first Hill, i immediately ac- counted thus: That Head, or Ear, was very large, and stood prominent from the Plant, pointing with its Silk Westward directly to- ward the next Hill of Indian Corn; and the Farina, I know, when very ripe, on shaking the Stalk, will fly off in the finest Dust, somewhat like Smoak. I therefore, with good Reason, judged that a Westerly Wind had wafted some few of these Particles from the other Hill, which had light on the Stiles of this Ear, in a Situation perfectly well fitted to receive them, which none of the other Ears, on the same Hill, had. And in- deed I admire that there were not more of the same Ear than I found impregnated in the same manner. As I was very exact in this Experiment, and curious enough in my Observations, and Zirkle: Early Records of Plant Hybrids 443 COLONIAL GOVERNOR AND EXPERIMENTALIST James Logan was among the first to put to the test of experiments the many con- jectures regarding sex in plants which were current in the early Eighteenth Lentury. By isolating maize and protecting the silks from pollen, Logan showed that larma from the maize tassel was necessary to produce viable seed. this, as I have related it, is truly Fact, 1 think it may reasonably be allowed, that not- withstanding what M. Geoffroy may have delivered of his Trails on the same Plant. I am positive, by my Experiment on those Heads, that the Silk was taken quite away, and those that were cover'd with Muslin, none of the Grains will grow up to their Size, when prevented of receiving the Fa- rina to impregnate them, but appear, when the Ears of Corn are disclosed, with all the Beds of the Seeds, or Grains, in their Ranges, with only a dry Skin on each, about the same Size as when the little tender Ears appear fill'd with milky Juice before it puts out its Silk. But the few Grains that were grown on the single Ear, were as full and as fair as any I had seen, the Places of all the rest had only dry empty Pellicles, as 1 have described them; and I much question whether the same does not hold generally in the whole Course of Vegetation, though, agreeable to what I first hinted, it may not be safe to pronounce absolutely upon it, without a great Variety of Experiments on different Subjects. But I believe there are few Plants that will afford so fine an Op- portunity of observing on them as the Mayze, or our Indian Corn; because its Stiles may be taken off or left on the Ear, in any Proportion, and the Grains be after- wards numbered in the Manner I have men- tioned. John Bartram I aiB indebted to Professor Rodney H. True for calling my attention to the following letters collected by V^illiam Darlington and included in his book, Memorials of John Bartram and Hum- phry Marshall, with Notices of their Botanical Contemporaries, published in Philadelphia in 1849. Darlington, un- fortunately, did not always print the letters in full and often omitted just the passages which today we consider the most important. Most of these letters are now in the possession of the Penn- sylvania Historical Society and I have been able to supplement Darlington's selections from his original source. I have not, however, been able to find what appears to be the crucial letter written by John Bartram to Peter Col- linson dated April 29, 1740. An undated Ms. in Bartram's handwriting probably includes the essential experiments de- scribed in this letter. It may even be a fragment of the letter itself, or more probably a rough draft from which the letter was copied. The writers and re- cipients of the letters were : 1. James Logan, "President of the Council, and Chief Justice of the Prov- ince of Pensilvania." 2. John Bartram, the celebrated Philadelphia Botanist; first to establish a Botanical Garden in America for the cultivation of both native and exotic vegetation. 3. Peter CoUinson, F. R. S. and F. S. A., London merchant and Quaker, an importer of plants. 4. Colonel William Byrd of Virginia, explorer, author and founder of the City of Richmond. 1736. James Logan to John Bartram (Darlington, p. 307) Friend J. Bartram: Last night, in the twilight, 1 received the inclosed, and opened it by mistake. Last year PETER sent me some tables, which I never examined till since I last saw thee. They are six very large sheets, in which the author (Linnaeus) digests all the productions of Nature in classes. Two of them he be- stows on the inanimate, as Stones, Minerals, Earths; two more on Vegetables, and the other two on Animals. His method in the Vegetables is altogether new, for he takes all his distinctions from the stamina and styles, the first of which he calls husbands, and the other wives. He ranges them, therefore, under those of 1 husband, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 20, and then of many hus- bands. He further distinguishes by the styles, and has many heads, under which he reduces all known plants. The performance is very curious, and at this time worth thy notice. I would send it to thee, but being in Latin, it will want some explanation, which, after I have given thee, thou wilt, 1 believe, be fully able to deal with it thyself, since thou generally knows the plants' names. If thou wilt step to town tomorrow, thou wilt find me there with them, at E. SHIPPEN'S, or J. PEM- BERTON'S, from 12 to 3. I want also to- 444 The Journal of Heredity Zirkle: Early Records of Plant Hybrids 445 say something further to thee, on micro- scopical observations. Thy real friend, J. Logan. Stenton, 19th of June, 1736. 1737. Peter Collinson to John Bartram (Excerpt, Darlington, p. 107) London, December 20, 1737. Dear Friend: I shall now consider the remaining part of thine of July 19. The magic lantern is a contrivance to make sport with ignorant people. There is nothing extraordinary in it, so not worth thy further inquiry. Thee art still desirous of a magnifier for flowers. Pray make this complaint to J. LOGAN, and try his thoughts. As thy inquiries seem in some measure to be owing to him, and as thee art his pupil (which no man need be ashamed of), no doubt but he will furnish thee with suitable instruments for that purpose, in order to render thy dis- coveries more perfect — so undoubtedly more to his satisfaction. . . . The references in the following to Dr. Tschiffely and to Byrd's letter of March 23rd enable us to place Bartram's com- munication in 1739. The greater part of Bartram's letter has been published (Darlington, 1849). The most impor- tant portions, however, were omitted by the modest Darlington. These I have italicised as they contain one of the passages which enable us to identify the genus investigated by Bartram as Lychnis, 1739. John Bartram to Col. Wm. Byrd (Bartram Papers, Vol. I, fol. 19) Dear Friend Coll Byrd: I r'cived thy kind letter by ye post last winter; and another dated March ye 23d, which I received by ye hand of thy friend doctor Tschiffely, whom I received very kindly and made as welcome as my present Circumstance would afford for thy sake hav- ing no other acquaintance than thine and another recommendation. I have this spring made several microscopical observations upon ye malle and femall parts in vegetables to oblige some ingenious botanists in Leyden, -who requested that favour of mee which I hope I have performed to their satisfaction and as a mechanical demonstration of ye cer- taity of this hypothesis of ye different sex in all plants that hath come under my no- tice. / canl find that any vegetable hath fozver to f reduce ferfect seed able to frofa- gate without ye conjunction of malle seed any more than anim^als and by a good micro- scofe ye malle and fem^all organs is as flain- ly discovered. I have made several Suc- cessfull experiments of joyning several spe- cies of ye same genus whereby I have ob- tained curious mixed Colours in flowers never known before but this requires an ac- curate observation and judgment to know ye precise time when ye fem^all organs is disposed to receive ye Tuasculin seed and likewise when it is by ye masculin organs fully perfected for ejection, I hope by these practicall observations to open a gate into a very lar(g)e field of experimental knowledge which if judiciously improved may be a con- siderable addition to ye beauty of ye florists garden. The next letter from Peter Collinson to John Bartram has also been pub- lished by Darlington (1849) and again I have italicised the portion he omitted. 1740. Peter Collinson to John Bartram (Bartram Papers, Vol. II, fol. 53) London, July 23d, 1740. Dear Friend: I had the pleasure of thine, of April 29th, 1740. Thy experiment of the use- fulness of the Farina^ is very curious and en- tertaining, where plants of a class are grow- ing near together, they will mix and pro- duce a mingled species. An instance we have in our gardens, raised by the late THOMAS FAIRCHILD, who had a plant from seed, that was compounded of the Carnation and Sweet William. It has the leaves of the first, and its flowers double like the Carnation — the size of a Pink — but in clusters like the Sweet William. It is named a Mule — per analogy to the mule produced from the Horse and Ass. Writing on these matters, brings to mind the Papaw — an Indian fruit — which in our stoves is produced in great plenty. On this tree, is very remarkably distinct, male, fe- male, and hermaphrodite blossoms, which are very extraordinary to see: but whether the last is an assistant in generation, or is a sport in nature, is not yet agreed .... Dr. WITT'S hollow-leafed Lavender, is, no doubt, the Side-saddle flower; but what relation it has to Lavender^ I must leave to him. The plant with tricolor leaves, I am well assured, is your fine Clinofodium, Our late severe winter has carried all mine off"; so pray send me some more seed — and of the Lychnis with Crosswort laves .... The Flesh Colour*^ Lychnis does not affear but in its Roome One with a fale Blew Flower fhafs this m,ay be that from Susquehannah a new one and very sweet sented which I esteeme much — and I want when thee in- creases it^ that with a zvhite flower with a Red Sfott in the Center^ and that Lychnis with a small white flower .... What curious experiments did Bar- tram recount in his letter of April 29, 1740, which caused Collinson to com- ment, ''When plants of a class are growing near together, they will mix and produce a mingled species," and to follow the comment by a description of the then twenty-three year old hybridi- zation experiment of Thomas Fairchild ? The inference is obvious: i. e., that Bartram described his own work on species crossing, the same work to which he referred in his letter to Colonel William Byrd. The following is transcribed from an undated Ms. in Bartram's handwriting which is now in the possession of the Pennsylvania Historical Society (Bar- tram Papers, Vol. I, fol. 20). The Ms., consisting of a frayed sheet of paper with a large fragment torn from a cor- ner, shows signs of being very hastily written, i. e., words inserted between the lines, words duplicated, letters small and less clearly formed than usual. The paper had been torn before Bartram wrote on it, so it seems improbable that he ever intended to include it in a let- ter. The Ms. has the appearance rather of a preliminary draft, possibly of the missing letter to Collinson. .... have had for 3 year a lychnis which produced flesh colored flowers male female upon several plants, one large plant which produced malle over most part of summer but produced no seed which ingaged my perticular notice and I observed that that seed was produced which ye sent me distinct male & female plants a year after I had sent from eng(land) some of ye seed of white lichnic which produced male & female flow- ers distinct upon different plants. It hap- ened that one female plant flowered neare two weeks before any male flowered which I obsrved daly and observed that tho there was no stamina or anthera in ye flower yet ye capsula was filed with perfect seed which I sowed as soon as it was ripe which came up very well in a few days but by that time that ye capsula of this first flowering white lychnis was near full grown then several malle plants flowered in ye same bed. I then puled of all ye capsulas that was set before any of these male flowers of ye white lic(hnis) appeared & that which was pro- duced after ye male blosoms opened and shed their farina: for I concluded that either that this female white lychnis must be impregnated with ye male red lychnis which grewde 10 yards of it if so then it must pertake of ye nature & color of ye red one or else we should be pusled to reconcile ye ye hypothesis of nescesity of ye male & female parts as after it had happened ac- cording to my expectation for it produced flowers great deal paler then ye red & as much higher coloured then ye white. But ye seed of ye white lychnis that was pro- duced after ye male plant flowered produced plants which bore flowers as white as thair original. Moreover I gave doer witt one plant of ye female lychnic which flowered plentifuly with him & produced ye capisula but containing onely something like ye husks of ye seed but no vegitive life on them, by which it apears that ye male parts of vegi- tables is realy as nexcesary to vegitation. Chronological List of Important Con- tributions to Our Knowledge of Hybridization and Sex in Plants, 1676-1761 The following table lists the work of those botanists who discovered by ob- servation and experiment that plants re- *Not skin-color. In 1740 "flesh-colour" melon. meant light red, like the "meat" of the water- 446 The Journal of Heredity produced sexually and that it was pos- sible to cross different varieties and spe- cies and secure new forms. It is easier to evaluate the contributions of Cotton Mather, Paul Dudley and John Bartram and the re-dated work of Philip Miller and James Logan by placing them in chronological sequence with the inves- tigations of the other contributors. 1676 — Grew in an address before the Royal Society, recorded his own belief and that of Sir Thomas Millington, that the stamens are the male organs of the plant and that the pollen acts as vegetable sperm. 1694 — Camerarius In a letter to Valentin describes his experiments which proved that pollen is necessary for seed development In Mercurialis, Morus, Rianus and Zea Mays. 1703 — Moreland tried to discover how pol- len Influenced the ovules. 1714 — Geofl"roy reported that the seeds of Zea mays and Mercurialis are infer- tile when no pollen reaches the stigma. 1716 — Cotton Mather reported certain ob- servations on Zea Mays and recorded (1) wind pollination, (2) hybridi- zation (variety cross) and (3) the resemblance of some of the progeny to the male parent; he also reported a cross ( ? ) between Cucurbita Pefo and C. maxima ( ? ) 1717 — Bradley removed the anthers from Tulips, which consequently produced no seed. He also noted spontaneous hvbrldlzatlon In several varieties of Pyrus Malus and Auricula {Primula A uricula) , 1717 — Fairchlld crossed Dianthus caryo- fhyllus with D. barbatus and found that the progeny resembled both par- ents. This was the first recorded ar- tificial hybrid. 1721 — Miller described (1) several Instances of spontaneous hybridization in Bras- sica and the resulting mixed and var- iable progeny; (2) Insect pollina- tion In Tulip; and (3) sexual re- production In Sfinacio and Cucumis. 1723 — In a garden In Stenbrohuld, accord- ing to Linnaeus, all male flowers were removed from a plant of Cu- curbita Pefo which, consequently, bore no fruit. 1724 — Dudley described wind pollination and variety crosses in Zea Mays, 1735 — Linnaeus commented on the artifices resorted to by gardeners to obtain hybrids In Tulifa and Brassica, 1735 — Logan showed by controlled experi- ments that pollen Is necessary' for the formation of grain In Zea Mays. 1739 — Bartram, in a letter to Col. Wm. Byrd, states that he has crossed spe- cies {Lychnis), obtained strange hy- brid forms and hoped to develop new horticultural varieties. 1749 — Gleditsch fertilized a female palm tree {Chamaerofs humilis\ ) In Ber- lin with pollen brought from a male tree in Leipzig, whereupon the palm produced fertile seed for the first time. 1750 — Haartman records forms which he assumes on taxonomic grounds to be natural hybrids in the genera Ver- onicay Delfhinium,, Safonaria, Ac- taea, etc. 1761 — Koelreuter systematically investigated hybridization and produced artificial hybrids in Nicoliana, Kedmia, Dian- thuSy MatthioUy HyoscyamuSy etc. Summary A number of early records of plant hybridization made by English and American botanists have been unknown to the historians of the science, while others have been referred to in a man- ner which minimized their importance. Certain other observations which helped to prove that plants reproduced sexually have been recorded, but mis-dated. L The first record we have of spon- taneous hybridization is a letter written by Cotton Mather on September 24, 1716, to James Petiver, F. R. S. The account was printed by Mather in Re- ligio Philosophica: or The Christian Philosopher, London, 172L He de- scribed variety crossing in Zea Mays and the pollination of squash by gourds. 2. The first recorded artificial hybrid resulted from a cross, Dianthus caryo- phyllus X D. barbatus. The investiga- Zirkle: Early Records of Plant Hybrids 447 tor who made this cross should be known as Mr. Thomas Fairchild, of Hoxton, not as "a gardener of Lon- don.*' Fairchild's hybrid is described by Bradley (1717), Miller (1731), Collin- son (1740), and Darwin (1781). 3. Philip Miller was the first to de- scribe insect pollination (in tulips). The date of his observation was neither 1751 (Sachs) nor 1731 (Green), but was at least as early as 1721. In that year he wrote to Bradley (Oct. 6) and Blair (Oct. 19 and Nov. 11) and de- scribed not only insect pollination in tulips and sexual reproduction in spin- ach, cucumbers and melons, but also spontaneous hybridization in cabbage when different varieties were grown near one another. 4. Paul Dudley in 1724 described wind pollination and variety crosses in Zea Mays. 5. James Logan in a letter to Peter CoUinson dated November 20, 1735, de- scribed his famous experiments on pol- lination in Zea Mays. He later pub- lished the results of his investigations in Leiden (1739) and London (1747). 6. John Bartram stated in a letter written in 1739 to Col. Wm. Byrd that he had crossed species within a genus and obtained a mixed and variable hy- brid progeny. An undated manuscript in Bartram's handwriting contains an account of hybridization in Lychnis. Bibliography 1. Blair, Patrick. Botanick Essays, Lon- don, 1720. 2. -. Observation upon the Gen- eration of Plants, etc., Phil. Trans. Roy. Soc. London, 31:215-24. 1721. 3. Bradley, Richard. New Improve- ments of Planting and Gardening, both Philosophical and Practical, explaining the motion of Sap and Generation of Plants, London, 1717. 4. . A Philosophical Account of the Works of Nature, London, 1721. 5. . A general treatise on hus- bandry and gardening, 2 vols., London, 1726. 6. Britten, James and Boulger, G. S. A Biographical Index of British and Irish Botanists, London, 1893. 7. Camerarius, R. J. De Sexu Plantar- urn, Epistola, Tubingen, 1694. Also in Ost. Klass exak Wiss, No. 105, Leipsig, 1899. 8. Darlington, Wm. Memorials of John Bartram and Humphry Marshall, Philadel- phia, 1849. . ^ 9. Darwin, Erasmus. The Botanic Gar- den. A Poem in two parts. Part II, The Loves of the Plants, 2nd Amer. Ed., New York, 1807. 10. Dudley, Paul. Observations on some of the Plants in New England, with remark- able Instances of the Nature and Power of Vegetation, Phil. Trans. Roy. Soc. London, 33:194-200. 1724. ^ ^ 11. Ellis, Sir Henry. History and An- tiquities of the Parish of Saint Leonard, Shoreditch, London, 1798. 12. Fairchild. Thomas: The City Gar- dener, London, 1722. 13. . An Account of same new Experiments, relating to the different, and sometimes contrary Motion of the Sap m Plants and Trees. Phil. Trans. Roy. Soc, London, 33:127-129. 1724. 14. Felton, S.: Portraits of English Au- thors on Gardening, 2nd Ed., London, 1830. 15. FocKE, WiLHELM Olbers : Pflanzen- mischlinge, ein Beitrag zur Biologic der Gewachse, Berlin, 1881. 16. Green, J. Reynolds: A History of Botany in the United Kingdom, London, 1914. 17. Grew, Nehemiah: Anatomy of Plants, London, 1682. 18. Harshberger, J. W.: James Logan, an early contributor to the doctrine of sex in plants, Bot. Gas., 19:307-312. 1894. ^ 19. KiTTRiDGE, G. L.: Cotton Mathers Scientific Contributions to the Royal Society, Amer. Antiquarian Soc. Proc, 26 : 18-57. 1916. 20. Koelreuter, J. G. : Volaufige Nach- tricht von einigen das Geschlecht der Pflan- zen, Leipzig, 1761, '63, '64, '66; also in Ost. Klass. exak. IViss. No. 41, Leipsig, 1893. ^ . ^ 21. Logan, James: Some Expenments concerning the Impregnation of the Seeds of Plants. Phil. Trans. Roy. Soc. London, 39:192-195. 1736. ^ ^ ., 22. . Experiments and Consid- erations on the Generation of Plants, London, 1747. 23 Mather, Cotton: Religio Philoso- phical or. The Christian Philosopher, Lon- don, 1721. ^ , >T 1 .L 24 MiALL, L. C. : The Early Naturalists, their Lives and Works (1530-1789), London, 1912 25! MaLER, Philip: Letter to Mr. Rich- ard Bradley dated Oct. 6, 1721. Published bv Bradley in "Treatise of Husbandry and Gardening," Vol. I, 330-332. London^ 1726. 26. . The Gardener's Diction- arv : 1st Octavo Ed., London, 1724 1st Folio Ed.. London, 1731. 2nd Folio Ed., London, 1733. 4th Abridged Ed., London, 1754. 7th Folio Ed., London, 1759. 5th Abridged Ed., London, 1763. 448 The Journal of Heredity 9th Folio Ed., edited by Thomas Mar- tyn, London, 1807. 27. Nichols, John : Literary History of the Eighteenth Century, 8 vols., London, 1817-185L 28. Pfeffer, W.: Ostw, Klcuss, cxak Wiss, No. 41, 1893. 29. PuLTENEY, Richard: Historical and Biographical Sketches of the Progress of Botany in England, 2 vols., London, 1790. 30. Roberts, H. F. : Plant Hybridization before Mendel, Princeton, 1929. 31. Sachs, Julius von: Geschichte der Botanik von 16. Jahrhundert bis I860,. Munich, 1875. 32. History of Botany (Au- thorized translation by H. E. F, Garnsey),. Oxford, 1890. VACUOLES IN PRIMARY MERISTEMS. By Conway Zibkle. Department of Botany, University of Pennsylvania. With Plates I— IV. ( Eingegangen am 6. April 1932.) During the middle of the nineteenth century the belief arose that cytoplasmic accumulation of water in quantities sufficient to form visible droplets causes the formation of vacuoles de novo. Undifferentiated or meristematic cells were held to be non- vacuolate and vacuoles were considered to develop as concomitants of cell enlargement and tissue differentiation. Between 1885 and 1890 a series of papers appeared in which generally accepted views were challenged. De Vries (48) argued that vacuoles develop from plastid-like bodies, tonplasts, in the apical meri- stems. Van Tieghem (47) accepted de vries' conclusion, but renamed the tonoplasts, hydroleucites. Went (49) , having demonstrated the presence of vacuoles in the reproductive cells of algae and in tissues where their presence had hitherto been unsuspected, came to the broad conclusion that *'all plant cells contain vacuoles" and that "all vacuoles arise by the division of pre-existing vacuoles and never originate de novo.'' Unfortunately the methods used by these investigators in demon- strating the presence of vacuoles were open to criticism. The technique of DE Vries and of Went for rendering vacuoles distinct was to place cells in a 10 to 15% solution of potassium nitrate. As Klebs (29) very emphatically stated cells so treated are not normal. Furthermore, Went's demonstration of vacuoles in cells of marine algae by placing the plants in distilled water was unsound. For this reason, his observations on Uving, untreated cells were ignored, and when Pfeffer (40) experi- mentally caused vacuoles to arise de novo in the plasmodium of a Myxomy- cete by introducing into it soluble granules of Asparagin, the tonoplast theory was generally held to be untenable. Thus, meristems are figured and described as nonvacuolate in most botanical text-books. The work of the Dangeards (8) and of Guilliermond (19 — 22) has again focused attention upon the origin and development of vacuoles. These investigators maintain that typical liquid vacuoles of differentiated tissues arise by the hydration of minute, viscous bodies resembUng Conway Zirkle: Vacuoles in primary Meristems. 27 mitochondria which occur characteristically in meristems. They classify the unhydrated, mitochondria-like bodies as vacuoles and conclude that all plant cells are vacuolated. Such a modification of the classical conception of the plant vacuole is not accepted by Priestley (41) who concludes from physiological evidence that there are three distinct types of cells in the stem growing point: (1) the meristematic cell, (2) the vacuolating dividing cell and (3) the vacuolated extending cell. He states that no vacuoles are recognizable in the meristematic cell. Bailey (2) working with living tissue cells has recently shown that the initials of the cambium are vacuolated. Indeed, certain of them are as highly vacuolated as are plant hairs. He finds a striking seasonal variation in the size, shape and number of vacuoles. In many plants the volume of vacuolar material is greatest during the period of rapid cell division. In the case of the secondary tissues the vacuoles obviously do not develop by the hydration of minute, viscous mitochondria -like bodies. The establishment of these facts has made it desirable to reinvestigate root tips, stem growing points and bases of embryonic leaves, to determine, if possible, whether there are fundamental structm-al differences between terminal and lateral meristems. In studying the distribution and development of vacuoles in different tjrpes of primary meristems and in their differentiating derivates, the writer selected the following plants for investigation: Pinus strobus, Robinia Pseudo-Acacia, Phaseolus vulgaris, Polygonum sachalinense, Fraxinus americana, Zea Mays, Osmunda Claytoniana, and Lumularia vulgaris. As the smaller vacuoles are completely destroyed by the usual cytological methods, it was essential to perfect new techniques for preserving the vacuoles in as unmodified a condition as possible. The work was done in collaboration with Professor Bailey, who made and examined living preparations of the several growing points. Thus, it was possible to evaluate the different fixatives by comparing the various fixation images with the visible structure of the living cells. Fixation of Vacuoles. Ten years before Golgi (16, 17) reduced silver in the "reticular apparatus" and thus discovered the structure which bears his name, BoKORNY (4) reduced silver in the vacuoles of Spirogyra from a very weak solution (1 : 100000) of ammoniacal silver nitrate. Three years previous to this, de Vries (48) reduced osmium in plant vacuoles from a 1% solution of osmium tetroxide in 10% potassium nitrate. It is inter- esting to note here that the essentials of the two most usual methods for demonstrating the Golgi apparatus in animal cells were first used to '^ stain" the plant vacuole. Bokorny interpreted the reaction as indicating the presence of an active (reducing?) protein in the cell sap and for this 1 28 Conway Zirkle: reason classified the proteins into two groups, active and inactive. Klemm (30) objected to Bokornys interpretation and very rightly pointed out that before the presence of an "active" protein could be demonstrated, it would first be necessary to prove that the vacuoles contained no tannin or other reducing agent. De Vries considered the reduction of osmium tetroxide to be due to tannin. He also discovered that vacuoles were fixed with salts of other heavy metals: HgClg, AgNOg, CUSO4, etc. Loew and Bokorny (32) showed that the contents of the vacuoles were precipitated in a granular form by bases. The usual cjrtological prepara- tions fixed by acid showed the vacuoles as hollow spaces. In describing the fixation images of the vacuoles it is essential to use certain terms in a very rest icted sense. As the vacuoles are so intima- tely associated with the cytoplasm, a cell in which the latter is accura- tely preserved will show a very clear "negative" of the vacuoles. This is the easiest method of maintaining the form of the vacuoles, except in those cases where their contents form insoluble compounds with certain heavy metals, — iron, copper, chromium, nickel, etc. In general, emphasis is here placed upon the fixation of the cytoplasm in such a manner as to have its observable mass and Umits unaltered. In describing the images it is convenient to use two terms which have been employed in con- nection with the alveolar theory of cytoplasmic structure. When the cytoplasm is fixed as a smooth mass with clearly defined limits but with no visible micellae of fibrils, it will be called hyaloplasm; when it is fixed as a reticulum or as tangled strings, the typical acetic acid fixation simage, it will be referred to as spongioplasm. Whether hyaloplasm and spongioplasm are different substances or different fixation images of ihe same substance is at present of no consequence. Whenever reference is made to a clear, limiting membrane about a vacuole, it will be called a tonoplast without any commitment to the hypothesis of de Vries that vacuoles evolve from plast id-like bodies. When root tips or stem growing points are treated with the usual nuclear fixatives, the cytoplasm appears as spongioplasm. The amount of stainable material in the tangled strands which form a crude reticulum is roughly proportional to the amount of cytoplasm in the cell. It has been a biological convention to illustrate this cjrtoplasm by stippUng in a reticulum and leaving colorless the numerous "vacuoles" bounded by the various strands. Sometimes the strands of spongioplasm seem to permeate a single large vacuole. Nothing resembUng these vacuoles exists in the Uving cell. They are fixation artifacts, and Hardy (23) has produced them by treating known homogeneous substances with the fixatives in question. These artifical vacuoles are produced when the cell is fixed with chromic acid or with bichromates on the acid side of Pjj 4,2 (Zirkle, 52). They are also produced by acetic acid and by both acid and basic acetates (Zirkle, 53). All of the lower fatty acids Vacuoles in primary Meristems. 29 and their salts cause this same artifact, even when combined with chro- mates or formaldehyde. In so far as the cytoplasm is concerned there is no observable difference between the fixation of formic, acetic, propionic, butyric, and valeric acids, and their copper and nickel salts. The mitochondrial fixatives, on the other hand, give a much more accurate image of the vacuoles (Bensley, 3). Bichromates on the basic side of the range pn 4.2 to pn 5.2, depending on the specific cation, fix the hyaloplasm and preserve the true vacuoles, though not always in the exact form that they have in the Uving cell. There is a tendency for the cytoplasm to shrink slightly and to acquire a granular boundary. It also appears a trifle thin and washed out and is not well mordanted. These faults of bichromate fixation can be overcome by the addition of formaldehyde to the mixtiu-e, although by so doing, additional compli- cations are introduced. The fixative is very unstable, as the formalin reduces the bichromate. In order to prevent the reaction from taking place too rapidly, it is necessary to make the solution quite dilute ; the mixture should never contain more than 4% formalin and 3% bichromate. As a rule, the more basic the bichromate, the more slowly it is reduced. The most successful mixture used is: K2Cr206 ^-^^^ (NH,)A20, l-2f^^^ CUSO4 1-^g Formalin (40% Formaldehyde) 10.00 c.c. Water 90.00c.e. The formalin is, of course, added immediately before fixation, which should be completed in 48 hoiu^s. The disadvantages of using an unstable fixing fluid can be obviated if the chromium is added in a reduced form, — i. e., as a chromic salt. Chromic sulphate Cr2(S04)3-15 H^O gave the best results. Formalm can be used with it in any desired concentration. Good results were obtained by the formula: Cr2( 804)3. 15 H2O l^ ^^Q * Slight excess Formalm*(40% Formaldehyde) i^'^r^n^'*'* ^ . 90.00 to 50.00 c.c. The copper oxide brings the Ph of the mixture to 4.6 The best concentration of formaUn depends upon the material to be fixed and must be determined empirically. Too great a concentration causes some plasmolysis and a swelling of the vacuoles; if too little is used, it is so diluted by the contents of the tissue that the mixture gives an erratic fixation image. ,.,,..• j t- Even at their best, however, these mitochondrial fixatives do not preserve the vacuoles unaltered. The tonoplast, so conspicuous m 30 Conway Zirkle; Vacuoles in primary Meristems. 31 living cells, is completely destroyed, and the vacuoles are delineated only by the fixed, faintly granular hyaloplasm. No reliable fixative for the tonoplast has yet been found. Although it can be fixed by many different mixtures, unfortunately they distort the shape of the vacuoles. This erratic behavior of the tonoplasts to fixation is doubtless due to some uncontrolled factor, either in the cell or in the fixative. A list of the mixtures which have preserved the tonoplasts may be of interest in itself, even though the mixtures themselves have little in common. The tono- plasts have thus far been fixed with (1) a 2% solution of trichloracetic acid ; (2) a mixture of 1% formic acid and 4% formaldehyde; (3) a solution of 3 % propionic acid ; (4) a mixture of 5 % copper butyrate and 4 % f ormaHn ; and (5) chrome alum titrated to pg 4.6 with ammonium hydroxide. While it is relatively simple to compare fixation images with sections of living tissue wherein the cells may be kept alive for several hours, it is not so easy to prove that the vacuoles are hot abnormal. Nemec (34) has recorded the extreme vacuolization that follows wounding. Checking the fixation image in the epidermis of very small roots does not meet with this difficulty, as here cutting is not necessary. Differences in the fixation image of the peripheral and central regions of root tips have been recorded for several fixing fluids (Zibkle, 52). When treated with the mixture of chromium salts and formaldehyde, however, the entire root tip, except in the cases described in the text of this paper, has a uniform fixation image which is essentially like the uninjured cells in sections of living tissue immediately after they are cut. This image is also like the epidermal layer of living unsectioned tips. The vacuolization due to injury, noted by Nemec, does not occur immediately, but follows the cutting of the section with a definite lag. The fact that certain vacuoles contain tannins ^ makes it possible to fix them directly and not as negative images in the cytoplasm. The iron salts are very valuable in this connection, as Dangeard (9) has pointed out. Cationic chromium and copper can also be used ; but the iron com- pounds, by forming conspicuous blue-black or green-black precipitates, obviate the necessity of subsequent staining (Fig. 1). The iron salts are far superior to the silver or osmium compounds for blackening tannin- filled vacuoles, as the reaction is more nearly specific than that of the other two heavy metals. The silver precipitate may be caused by a chloride in the vacuole and the subsequent exposure of the specimen to light. An osmium precipitate may indicate no more than the presence of an oil globule. In fact, osmium tetroxide is reduced to an insoluble black precipitate by so many different substances that it is very dangerous to draw any inferences from its abihtiy to blacken various cell organs. ^ The word "tannin" is here used in its most general sense. It includes all compounds containing aromatic substances which form green-black or blue-black prcsipitates with iron salts. Fortunately, some of the chemicals used as specific tests for the tannins can be used as fixing fluids. Thus, formaldehyde acidified with a little hydrochloric acid precipitates the so-called catechol tannins and is a fair fixing agent. The iron chlorides and sulphates, when combined with formaldehyde, fix the cells with very little distortion. It is necessary to take the vacuolar contents into consideration in any study of fixation. For example, when the root tips of Pinus are fixed with a mixture of potassium and ammonium bichromates, a fixative which normally destroys all chromatin, enough tannic acid is released from the vacuoles and added to the mixture to give certain cells a typical tannic acid fixation image. The chromosomes are well preserved. Before the mitochondria can be demonstrated in these cells, enough alkali must be added to the fixative to neutralize the acid contents of the vacuoles. It would be well to emphasize here certain technical difficulties in investigating both the size of the vacuoles and the nucleo-cytoplasmic ratio in all root tips of the Zea type. The vacuolar fixatives penetrate the calyptrogen very slowly and the fixation image is apt to vary greatly in different specimens, especially if the root cap has an outer gelatinous layer. Thus far, the only fixatives found which penetrate the tissues readily are those which contain some one of the lower fatty acids, and these fixatives, of course, destroy all details of the vacuoles. The cells, too, are under considerable tension in the living root and, during a rela- tively slow fixation, are apt to be disrupted completely by the changes in volume of the outer cell layers. Often the entire root tip has a fixation image which checks perfectly with the living structure, except for the apical cells of the dermatogen and periblem, which have collapsed. Obviously, any attempt to measure the nucleo-cytoplasmic ratio in these cells is very difficult, as there is almost certain to be a change in the cell volume with fixation. Those fixatives which seemingly cause the least change in cell-size, i. e., those which contain one of the lower fatty acids, destroy the vacuoles, i id, of couse, no nucleo-cytoplasmic ratio can be obtained if the cjrtoplasm contains an unknown volume of vacuolar material. Vacuoles in Initials and their Derivates. The plants investigated by the writer were selected as representing the commoner types of apical growth, both in the phanerogams and in the higher cryptogams. The cryptogams, Osmunda and Lunularia, elongate through the division of a single tetrahedral apical cell. The initial of Osmunda is relatively large (Fig. 16) and forms daughter cells on all four surfaces. The anterior daughter cell gives rise to the root cap, whereas the posterior daughter cells divide to form the root proper. In Lunularia, the initial is relatively small and divides in such a manner as to form daughter cells on the posterior surfaces only. 1 82 Conway Zirkle: These cells produce the thallus. Pinus has the typical gymnosperm root tip, a periblem which produces the root cap, epidermal layer and cortex, and a plerome which forms the central cylinder. Robinia and Phaseolus have but a single set of initials which cut off anteriory, cells to form the root cap, and posteriory, cells which constitute the root proper. In Polygonum, the dermatogen divides in two planes, the root cap arising through transverse, and the epidermis through longi- tudinal divisions. The rest of the root is derived from the periblem and plerome. Zea has the typical root tip of monocotyledons; distinct dermatogen, periblem and plerome which together give rise to the root, and a calyptogen which produces the root cap. Three recent papers have reported spherical vacuoles in the apical meristems of plants as far apart phylogenetically as the Muscineae (Motte 35), Selaginella (Emberger 13) and Elodea (GoNgALVES 18). Spherical vacuoles were also found in the promeristem of every root tip examined by the writer. Other forms of vacuoles also occur in this tissue but they seemed to be only temporary deviations from the spherical which may be considered the typical shape of these vacuoles. There are no traces in any initials of tonoplasts (in the sense originally used by de Vries), of mitochondria-like bodies which develop into vacuoles or of any other vacuolar primordia. In none of the primary meristems examined did vacuoles arise through the development of any other cell component hut regularly increased in number through the division of pre-existing vacuoles. As would be expected, there is considerable variation in the size of the vacuoles in the initial cells of the several species examined. In Pinus they range normally between 2 and 5 // in diameter. They are larger in Polygommi with diameters varying from 5 to 15 // (Fig. 14). The cytoplasm seems quite dense in the root initials of Robinia and Phaseolus and here the vacuoles are a trifle smaller on an average than those of Pinus. The vacuoles in the calyptogen and plerome of Zea are from 4 ^1 to 8 /^ in diameter and are somewhat smaller in the dermatogen and periblem. The vacuoles in the apical cell of the root tip of Osmunda, a cryptogam, are of an entirely different order of magnitude from those in the root tips of the phanerogams described above. This large tetrahedral cell (Fig. 16) has a volume comparable with the cambial initials of the Dicotyledons. It is so extremely vacuolate that sections ^ ft to 10 /i in thickness showed no cytoplasmic structure. Figure 16 represents a section 15 // thick. Here the cytoplasmic vacuolar ratio resembles that of the secondary meristem described by Bailey (2). The daughter cells also contain much more vacuolar material than cytoplasm. It seems very significant that the embryonic regions of certain relatively primitive plants are so highly vacuolate and are thus more like the secondary than the primary meristems of the Spermatophyta in their cell contents. Vacuoles in primary Meristems. 33 This might indicate that the cambial structure is not so highly specia- lized as it is generally considered to be. No account of the vacuoles in the root initials would be complete without a description of their extreme lability. In living sections they can be observed to elongate, fuse and again fragment. When there is cytoplasmic streaming, the vacuoles may at times be modified passively by the movement of the cell contents. In Pinus the smaller vacuoles sometimes partially fuse into chains, and- rarely, when the cells are in active cyclosis, they take the canalicular form. Under direct observations, spherical vacuoles in quiescent cells elongate, become filamentous and anastomose to form a reticulum upon the initiation of protoplasmic streaming. When streaming occurs in the meristematic initials of Poly- gonum, the vacuoles are also drawn out to form bead-like strings (Fig. 14). They are, however, always larger than Holmgren canals. In the initial of Robinia and Phaseolus vacuoles rarely, if ever, fuse to form canals although they are frequently distorted into prolate spheroids and hooked rods (Fig. 22). Zea generally has iso-diametric initials which regularly have only spherical vacuoles. Occasionally, the apical cells of the der- matogen and periblem become greatly elongated, whereupon their vacuoles become canal-like (Fig. 26). The vacuoles in the stem initials differ in no important aspect from those in the root tips. They are regularly spheroidal although at times they are distorted into various asymetrical forms. The stem growing point of Robinia is shown in Fig. 28, that of Fraxinus in Fig. 12. In the early spring the shoot apex of Polygonum is subterranean and may grow several inches through the ground before emerging to become a normal aerial shoot. The vacuoles in the apex are globular ranging from 5 /i to 15 // in diameter both before and after the stem reaches the surface. In Pinus there is an interesting difference between the vacuoles in the ini- tials of the long shoot (Fig. 11) and those in the short shoot (Fig. 13). In both, the vacuoles are spherical when separate. Frequently they are joined into irregular bead -like chains which are always too large to be mistaken for Holmgren canals. In the long shoot they fix just as do those in the root initials; on the contrary, in all three embryonic layers of the short shoot they contain tannin and fix as shown in Fig. 13. It should be recorded, however, that the specimens here described were obtained near the end of May, when the short shoot had completed its year's growth but while the long shoot was still rapidly elongating. In view of the large seasonal changes which occur in vacuoles (Bailey 2) we must grant the possibility of shoots fixed at another time of year showing a very different picture. The large fleshy rhizome of Osmunda has a very wide growing point The anterior laver is composed of a single, large, vacuolate, tetrahedral apical cell together with the daughter cells, which are themselves almost 3 Z. f . Zellforschung u. mikr. Anatomie. Bd. 16. 34 Conway Zirkle: as large and quite as vacuolate as the apical. Indeed as far as size and vacuolar contents are concerned, these cells are more comparable with the cambium than with the primary meristems of the phanerogams. The leaves of Osmunda, unlike those of the other plants investigated, elongate through apical accretions and have the same type of growing point as the root tips and rhizomes. Lunularia also grows through the activity of a single apical initial. The cell is tetrahedral like the apical cell of Osmunda but differs in that it is much smaller and contains rela- tively more cytoplasm. The vacuoles may be either sperical or, as is generally the case, drawn out into canals (Figs. 21, 23). HoF (25) has shown that the apical cell of Pteris has spherical vacuoles. Development of Vacuoles during Cellular Differentiation. The classical picture of the development of vacuoles in primary meristems shows, — (1) numerous small globular vacuoles originating in the zone of the cell division, well posterior to the initials ; (2) a gradual enlargement and fusion of vacuoles in the region of elongation; and (3), the final amalgamation of all vacuoles within a protoplast into the single large central vacuole typical of the fully differentiated plant cell. This concept of vacuolar development has been proven inadequate by the recent work of Guilliermond, the Dangeards, Pensa, Parat and their students. The workers have clearly demonstrated that certain vacuoles originate from primordia which, seemingly, have little in common with their mature forms. Great emphasis has been placed upon the super- ficial resemblance of these primordia to other cell inclusions such, for example, as mitochondria. Guilliermond (22) has traced the successive phases in the growth of the anthocyanin containing vacuoles in the young rose leaf. In the marginal cells they are filamentous, in the sub- marginal they thicken into rods, and finally swell into spheres and fuse to form the large central vacuole of the mature cell. All meristematic cells investigated by the writer were vacuolate and to this extent his findings substantiate the much earlier work of Went (49). The course of vacuolar development in the differentiating cells would thus begin in the promeristem. The sequence of developmental forms from the plerome to the pith follows the classical view very closely. Small globular vacuoles enlarge and fuse as the cells mature. The same stages occur in the vacuoles of the cortex and in nearly all iso-diametric cells in the embryonic regions of the phanerogams. Those embryonic cells, however, which pass through an elongated phase in the course of their development, contain a different series of vacuolar shapes. When they are first cut off from the initials their vacuoles are spherical, as they elongate their vacuoles tend to become filamentous, form Holmgre n canals and may even anastomose, producing a single reticulum. Examples Vacuoles in primary Meristems. 35 of such cells may be found in the epidermis of the root tip of Zea and in the procambial region of Zea, Pinus, Robinia, Phaseolus, and Polygonum. It is noteworthy that cytoplasmic streaming is much more active in these long cells than in the iso-diametric. In living sections spherical vacuoles can be observed to elongate into separate filaments and fuse into a composite network following the initiation of cyclosis. It is possible that the thread-Uke vacuoles owe their form to the distortion produced by the movements of the cell contents. The presence of reti- culate vacuoles in fixed preparations, however, is not an indication that cyclosis was active when the fixative reached the cell. In many cells which have dense cytoplasm, the reticulate form of the vacuoles persists long after the cytoplasm has become quiescent. Apparently the visco- sity either of the cytoplasm or of the vacuolar contents is great enough to prevent an immediate rounding up of the vacuoles. A reticulum may also be formed from the fusion of separate spherical vacuoles when there is an extremely slow movement of the cytoplasm. Cells which regularly possess reticulate vacuoles include the procambium of Polygonum (Fig. 15), Zea (Fig. 26) and Robinia (Fig. 23), and the cells in the base of the embryonic leaves of Pinus (Fig. 24). Filamentous vacuoles occasionally occur in certain root initials of Zea (Fig. 25) and more frequently in the apical cell of Lunularia (Fig. 21, 27). In general, though, the Holmgren canal is not the primitive form of the apical vacuole but only a temporary stage through which certain vacuoles pass. When cyclosis ceases or when the cell so increases in size and aqueous content that the vacuoles are not broken up or distorted by cytoplasmic streaming, they regain their spherical form, and, as the cell ages, fuse to form the typical large, central vacuole of the plant cell. Vacuolar development in the Osmunda root tip differs in detail from that in the phanerogams described above although the general sequence of homologous forms is parallel in the two types. The apical cell of Osmunda and its daughter cells contain numerous large vacuolos and but little cytoplasm (Fig. 16) and thus resemble the cambial cells described by Bailey (2). These large vacuoles are divided passively as the daughter cells divide and are further broken up by streaming cytoplasm. The small cubical cells derived ultimately from the daughter cells thus contain many small spherical vacuoles. As these latter cells differentiate and increase in size, their vacuoles enlarge and fuse. The evolution of vacuoles is here obviously from very large, through small to very large again. There is no question of vacuoles originating from plastid or mitochon- dria like bodies. The same sequence holds in the vacuolar development in the apex of the rhizome. The anterior layer of the thick stem growmg point is composed of a single apical cell surrounded by large vacuolate daughter cells. Back of this layer the cells and their contamed vacuoles grow progressively smaller until the small iso-diametric cells typical 3* t 36 Conway Zirkle: of most stem growing points is reached. On differentiating, the cells and vacuoles enlarge. While the apical cell of Lunularia is shaped like that of Osmunda, it differs in that it is much smaller and is filled with cytoplasm. The vacuoles are sometimes spherical although generally they are drawn out into canals (Fig. 21, 27). The daughter cells which also contain filamentous vacuoles give rise to cells of three types ; the epidermal cells, the peculiar palisade cells characteristic of the Marchantiales, and the mesophyll cells of the thallus. The development of vacuoles is essentially alike in all three kinds of cells and their sequence of forms resembles that described by GuiLLiERMOND in the young rose leaf. The gemmae bud off from a line of small undifferentiated cells extending directly from the apical cell. All contain small vacuoles either spherical or reticulate. Every cell of the gemma, during the entire embryonic development, contains vacuoles. As Lunularia reproduces exclusively by gemmae, there is here an instance of a complete, if somewhat abridged, life cycle in which vacuoles exist in all of the cells and apparently reproduce exclusively by the division of pre-existing vacuoles. Very little is known at present concerning the contents of the vacuoles in the primary meristem. The substances which can most easily be recog- nized definitely in the cell sap are the tannins and these are localized in cells somewhat removed from the initials. Most vacuoles can concentrate certain basic dyes (neutral red, brilliant cresyl blue, etc.) from very dilute solutions, although the vacuoles in the initials take the stains much less readily than do those which contain tannin. Several investigators have noted that different vacuoles assume very different colors when tested with a single dye and have interpreted this phenomenon as evidence of the existence of a special vacuolar substance with metachromatic properties. As the dyes in question are hydrogen ion indicators, this **metachromasy" may be due primarily to differences in pjj. Unfor- tunately the stain which is most reliable for vacuoles and which shows most clearly this color variation is neutral red, a mediocre indicator subject to relatively large salt and protein errors. The tannins form insoluble compounds with the fixatives used by the writer and consequently all tannin bearing vacuoles contain precipi- tates. In some instances precipitates are formed in vacuoles which contain no tannin. These occur, although rarely, in the root tip of Zea. Robinia and Phaseolus are likewise free from tannin, yet when their root tips are fixed with chromic sulphate they are rendered opaque throughout by a green precipitate, which extends even to the vacuoles of the initials (Fig. 22). At times there are precipitates in the vacuoles of the apical cell of Lunularia (Figs. 21, 27). The presence of such precipitates in meristematic initials may have been responsible for certain misconceptions concerning the origin of Vacuoles in primary Meristems. 37 oles. In accurately fixed specimens of Robinia and Lunularia these vacu- precipitates are clearly limited to the inner surface of the tonoplasts. When the vacuoles are filamentous the precipitates are of the size and shape of mitochondria. If the cells have been only slightly plasmolyzed the small vacuoles of the initials disappear leaving the precipitates which are easily mistaken for mitochondria. The differentiating cells still show both vacuoles and precipitates although with the size of the former somewhat reduced. Under these conditions it is possible to trace the development of vacuoles from mitochondria-like primordia, an error, almost unavoidable in investigating badly fixed material. The tannin containing cells in the stem apex of Pinus and Polygonum and in the underground rhizomes of Osmunda form very definite patterns, the significance of which is obscure. In the root tips of these plants the tannin is mainly confined to the peripheral regions leaving the central cylinder free except for a few isolated cells. The tannin filled vacuoles have a cytological importance aside from a consideration of any function they may serve. They have (1) essentially the fixing properties of the GoLGi apparatus in the animal cell ; (2) among them may be found all of the varied forms observed by Bailey (2) in the cambium and ray cells; and in addition (3) certain of them assume shapes possessed by no other vacuoles. Indeed some of these structures are so atypical that the general concept of the vacuole and its role in the economy of the cell will have to be broadened if they are to be included. It is evident that these aberrant vacuoles have sometimes been mistaken for various cytoplasmic inclusions which supposedly had no connection with the easily recognized vacuole of the mature plant cell. The root tip and embryonic leaf of Pinus contain representative samples of the tannin containing vacuoles and a description of the forms found there will show almost the complete range of variation so far found in these cell organs. Figure 1 represents a median longitudinal section through the root tip of Pinus, fixed with a mixture of formaUn and ferrous sulphate and unstained except for the precipitated iron tannate. The peripheral localization of tannin shows clearly. The only cells free from precipitate are the meristematic initals, the central cylinder and a core piercing the root cap and extending from the periblem to the apex of the root. At the anterior end of the central cylinder are two "barbs" which, while not entirely tanninless, contain too little to show at the magnification of Figure 1. These "barbs" contain numerous small spherical vacuoles which are faintly outUned in tannin. It would be well to emphasize that when cells containing but slight traces of tannin are fixed with the salts of the heavier metals, the insoluble tannates are always precipitated in the periphery of the vacuoles, the interiors appearing empty. The more I 38 Conway Zirkle: tannin present, the heavier the precipitate. In cross section these fixed vacuoles appear as rings, (Fig. 19) or occasionally as discs. It should be unnecessary to point out that when root tips are fixed whlth mixtures containing a heavy metal, the presence of rings, plates, rods, etc., is no indication that such structures exist in living cells. As might be expected, there is a great variation in the vacuoles in different specimens. Such factors as the temperature of the roots prior to fixation, the amount and availability of water and oxygen in the surrounding soil; etc., were not controlled, so the conditions necessary for the vacuoles to assume a given form are not known. In spite of a pre- sumptive variation in external conditions, however, the vacuoles in the several parts of the root tip tend to assume standard forms. Figure 19 shows cells in the anterior end of the periblem; perhaps the youngest in which tannin is recognizable. Typical of this region are elongated vacuoles which may be either straight or bent, and spherical vacuoles in which the tannin appears as a peripheral ring. Figure 10 shows numerous spherical vacuoles, arranged oftentimes in rows, which increase in size and fuse as the cells are pushed towards the exterior of the root. Figure 7 shows elongated cells nearer the surface of the root which, though nearly filled with tannin, retain it in separate tube-like vacuoles. Nearby cells contain vacuoles which are full of tannin, yet they seem entirely normal and are undergoing rapid cell division (Fig. 6). It seems remarkable that cells which contain so much tannin should be able to live. The fact that they do remain functional suggests that the tonoplast is able to keep the cytoplasm separate from the vacuolar contents ; otherwise the cell would be tanned. The last statement is not wholly hypothetical, for when the permeability of the cells is altered by numerous fixatives, such cells acquire the typical tannic acid fixation image. A trifle farther back, and adjoining the tanninless central region, a complete series of vacuoles are formed which show the effects of the amount of their contained tannin upon their fixation forms (Fig. 4). Next to the interior region they are merely outlined with a precipitate, then come transitional forms, and in the outer regions they are solidly black. In cross section some of these vacuoles appear as hollow circles completely surrounding the nucleus; in cell division the "circle" splits into two "semicircles" (Fig. 4), one going to each daughter cell. The division line between the tanninferous and tanninless region is often very abrupt (Fig. 3). Tannin-bearing vacuoles in the embryonic leaves of Pinus show an enormous variation in number, size, and shape. The most common form, found near the bases of the leaves, is a reticular apparatus (Figs. 18, 24). This form has been very accurately depicted by Dangeard (9). The reti- culum is quite delicate, and the strands are either ribbons or chains of tiny beadlike vacuoles (Fig. 18, 14). Sometimes larger vacuoles will Vacuoles in primary Meristems. 39 be joined by chains of smaller ones (Fig. 20). Farther along the leaf, the strands of the reticulum become much heavier (Figs. 8, 17), and at a point two to three millimeters from the stem apex these strands fuse to form a single, large, tannin-filled vacuole. The cells of the vascular bundles are nearly all free from tannin, but most of the other cells are well filled, particularly those which contain numerous chloroplasts. The Differentiated Core in certain Root Caps, In the root cap of Pinus there is a tannin-less core, four to five cells in diameter, extending through the tannin-bearing layer to the region of the most active cell division (Fig. 1). The fact that the cells surroun- ding the anterior portion of this core contain tannin serves to bring it into sharp relief when otherwise it might have escaped attention, for the cells in this core are differentiated in ways other than in their lack of tannin. If the root of Pinus is fixed as described and stained with Acid- Fuchsin, the core can be traced from the apex of the plerome to the exact tip of the root. Its anterior region can easily be differentiated from the siu-roimding tannin-bearing cells (Fig. 1), and its posterior region from the other cells of the periblem by the fact that it contains less cytoplasm and consequently is more lightly stained. It contains throughout the greater part of its length numerous large starch grains which presumably stimulate the geotropic responses of the root. There is no evidence, however, that this is the exclusive function of these differentiated cells, as the tannin-bearing cells are equally rich in starch grains. There is no possibility of interpreting this core as an artifact of fixation, as it is quite evident in living sections of the root tip stained with Neutral Red. The vacuoles in the core are much less acid than those filled with tannin found in the surrounding cells. While the core just described is more conspicuous in Pinus than in the other plants here investigated, is by no means limited to the Gymno- sperms. Is is very clearly shown in Polygonum sachalinense, as a zone containing very lightly staining cytoplasm which penetrates the peri- pheral tannin-bearing layers just as it does in Pinus (Fig. 2). It can also be shown to exist, though not so sharply delineated, in the root tip of the seedling of Robinia Pseudo-Acacia (Fig. 2). Here there are no clearly defined tannin-filled cells to mark its boundary, but it can be recognized as a region extending from the apex of the central cylinder to the tip of the root, the cells of which contain little cytoplasm. It is evident that the existence of this core in root tips is very widespread, although its distribution in the higher plants has not as yet been adequately vinestigated. It does not exist in the fern, Osmunda Chytoman^, al- though the peripheral tannin-bearing region is here as distinct as m Pinus. Nor does it exist in the Monocotyledon, Zea Mays, i 40 Conway Zirkle: |l: Discussion. The investigation of the several types of primary meristems here recor- ded has shown that the cells are, without exception, vacuolate. There is great variation in the size of the cells in different species, in the volume of the vacuoles, and also in the ratio of the volume of the vacuoles to that of the cells which contain them. Bailey (2) has shown that the cells of the secondary meristem are also vacuolated, and that the vacuoles have similar forms in both the cambium and apical growing regions. There seems to be no essential morphological difference between primary and secondary meristems except those attendant upon differences in cell size, the cambial cells being, in general, larger than those in root tips and stem growing points. While the larger cells may have larger nuclei and more cytoplasm than the smaller (Bailey, 1), the greater part of their size difference is due to the comparative volume of their vacuoles. Thus the vacuoles of the cambium are relatively much larger than those of most apical meristems. Certain plants, however, such as Osmunda, have large apical cells, the volume of which is close to that of many cambial cells. It is interesting to note that here the cytoplasmic vacuolar ratio is of the same order as that in the cambium, and that the mere presence of large vacuoles cannot serve to differentiate the secondary from the primary meristems. It cannot be urged in the case of Osmunda that the vacuoles in the growing points of the Archegonateae, where there is a distinct apical cell, are aberrant and not comparable with those in the Spermatophyta. When the apical cell is small, as in Lunularia, its vacuoles may resemble those in the primary meristem of higher plants. Thus large vacuoles in the primary meristem are not concomitant with the growing point derived from a single apical cell, but with the presence in the growing point of large apical and daughter cells (Fig. 16). There has been a tendency among morphologists to consider the apical growing regions undifferentiated, while the cambium has been looked upon as more or less highly specialized. If we assume that the ancestors of the higher plants were essentially like the simpler plants of to-day, it would follow that both primary and secondary meristems were derived from much more complex cells, and that their relative simpHcity is not primitive but has come about as consequence of their assuming the entire burden of growth. Both are specialized in that their cell inclusions are embryonic. It should be noted that the cambium, both in regard to cell size and vacuolate condition, resembles more closely the dividing cells of the Conjugateae and Chlorophyceae, and even certain apical cells of the Archegonateae, than do the small cells with dense cytoplasm of the primary meristem. The presence of vacuoles in vegetative cells is a factor which must be considered in all investigations of the nucleo-cytoplasmic ratio in Vacuoles in primary Meristems. 41 plants. The enormous variation in both nuclear and cell size in Pinus Strobus has been shown by Bailey (1). — Obviously, if the greater part of the volume of the larger cells is made up of vacuoles, a mere com- parison of nuclear size with total cell size gives no indication of the nucleo- cytoplasmic ratio. Nor can any estimates be made of the effective working sphere of the nucleus without a careful study of such factors as cyto- plasmic streaming and the cytoplasmic -vacuolar ratio. No evidence whatever was found indicating that vacuoles develop from tonoplasts, hydroleucites, mitochondria, or any other cytoplasmic inclusion. All of the cells examined contained vacuoles which continually changed their shape during cytoplasmic streaming, fused, and again fragmented. The only mode of origin that could be discovered for any individual vacuole was the division of a preexisting vacuole, and, to this extent, the work substantiates Went (49). No evidence was found of vacuoles originating de novo, which, of course, does not mean that this never occurs ; for it is quite possible that a new vacuole could be mistaken for a fragment of a dividing one in a cell whose contents are in a rapid motion. Guilliermond (21) has reported the neoformation of vacuoles in Saprolegnia. While it frequently happens that beaded, filiform, and rod-like vacuoles grow and become spherical and finally fuse to form the large central vacuole characteristic of most fully differentiated plant cells, this is not the usual developmental sequence. The vacuoles in the derma- togen, periblem, and plerome are regularly spherical and become thread- like or form Holmgren canals during cyclosis. This latter condition is apparently the rule only in the procambial cells, where, incidentally, the volume of the vacuoles in any one cell is much less than in a cell of the apex. The derivation of small vacuoles from large ones and their subsequent enlargement, which is so obvious in Osmunda, does not constitute an isolated instance. It is but a striking example of a general tendency. In recent years a number of cytologists, beginning with Bensley (3), have reported that the plant vacuole is homologous with the Golgi apparatus found in animals. If this conclusion holds, — and much evidence in favor of it has been brought to light by the investigations of Guilliermond (22), Dangeard (9), Parat et Painleve (38) and others, it would shed a great deal of light upon the Golgi apparatus, for, com- pared with this cell organ, our knowledge of vacuoles is very extensive and precise. Vacuoles have long been recognized as perfectly normal parts of living plant cells. Their behavior in Uving tissue and reaction to vital stains have been recorded by Guilliermond (19—22), Dangeard (9), Bailey (2), and others; their contents in certain cenocytic forms (Valonia, Nttella) have been analyzed by numerous workers, among others Brooks (5), Crozier (7), Hoagland and Davis (24), Irwin (28), f 42 Conway Zirkle: Vacuoles in primary Meristems. 43 OsTERHOUT (37), WoDEHOUSE (51). On the other hand, the ^'reticular apparatus" discovered by Golgi has been definitely recognized only in tissue which has been subjected to prolonged and complicated chemical treatments (Cowdry, 6). Parat and Painleve (38) have indeed found a reticular structure in living animal cells which stains with Neutral Red; and it is quite possible that this reticulum contains reducing sub- stances which react with silver or osmium salts to form the Golgi appa- ratus. The Golgi techniques depend primarily upon the reduction within the cell of some heavy metal, — generally silver or osmium. Chemically these tests are non-specific, and it is extremely difficult to make any precise inference concerning the nature of the apparatus from its reactions to the tests in question. Indeed, Ludford (33) defined the Golgi appa- ratus as " that region in the cytoplasm of cells which brings about the reduction of osmium tetroxide at a lower temperature, or in a shorter time than is required to produce a total blacking of the cell." If we accept the above definition, a tannin-filled vacuole would qualify as a Golgi apparatus, particularly if the vacuole were treated with osmium tetroxide when it had a reticular form (Figs. 8, 17, 24) ^. Such a ''reticular apparatus" in the base of the embryonic needles of Pinus has been described by Dangeard (9), who has identified it by much more specific tests than zoologists have applied to its possible homologue in the animal cell. But other inclusions besides tannin reduce osmium in the plant cell. If the reduction of osmium were the only criterion of the Golgi apparatus, oil droplets and lipoids would also qualify as would any cytoplasmic inclusion able to reduce osmium when the tetroxide is mixed with oxidizing agents such as potassium bichromate and chromic acid. Thus it is easy to understand why this osmium technique has given such capricious results when applied to plants. Obviously, much more work will have to be done on the animal cell and its reticulate metallic precipitates before the latter can be definitely identified as an animal vacuole. The distribution of tannin in certain root tips is strictly limited. Most of it is localized in a peripheral zone, while the central cylinder contains little or none. In Pinus and Polygonum the root cap is filled with tannin except for a differentiated core, about four cells in diameter, which extends from the tip of the root to the plerome. This core contains no tannin and is clearly distinct from its surrounding cells (Figs. 1, 2). There are, perhaps, relics of such a core in Robinia (Fig. 5) and Phaseolus. Eriksson (14) reported a distinct core in the root tip of Lupinus. No trace of such a core is visible in Zea or in Osmunda, although in the latter instance the root cap is filled with tannin. This core has quite a ^ It should be noted here that the reduction of osmium by the tannin filled vacuole is almost instantaneous, while Osmium is reduced by the animal Golgi apparatus very slowly. widespread phylogenetic distribution and would consequently be expected to serve some useful function. There is no evidence, as yet, as to what this function may be. As it is a tannin-free path leading from the region of most rapid cell division to the exterior of the root, it is strongly sug- gestive of a route for certain chemicals in the surrounding soil to reach the growing point without being conditioned by passing through tannin-filled vacuoles. Summary. 1. Vacuoles in the primary meristems of Pinus, Polygonum, Robinia, Phaseolus, Fraxinus, Zea, Osmunda, and Lunularia were investigated in both living and fixed preparations. 2. The usual acid fixing fluids completely destroy the true vacuoles and form artificial ones. Fixatives which contain acetic acid, any ace- tate, chromic acid, or a bichromate on the acid side of pn 4.2, give an entirely erroneous picture of the cytoplasm and Vacuoles. 3. When vacuoles contain tannin, they can be blackened with the Golgi techniques. They can also be fixed and stained by mixtiu-es of formaldehyde and iron salts. 4. Modifications of mitochondrial fixatives preserve the true form of the cytoplasm and vacuoles, in spite of the fact that the vacuolar mem- branes are destroyed. When the vacuoles contain no tannin, they appear as colorless regions clearly outUned by the faintly stained hyaloplasm. 5. Mixtures of ¥.^Qr^O^ and (NH4)2Cr207 with formaldehyde pene- trate well and preserve the size and shape of the vacuoles. However, the fluid is unstable. Equally good results were obtained by fixing the specimen with formaldehyde and Cr2(S04)3 brought to pn 4.6 with CuO. 6. All cells in the apical meristems contain vacuoles which are nor- mally spherical. On the initiation of cyclosis, however, these vacuoles may be drawn out into rods, threads, and even be made to anastomose to form a reticulum. Because of the viscosity of the cytoplasm, the canal- like or reticular form of the vacuoles may be retained some time after cytoplasmic streaming ceases. In the procambial cells the vacuoles are smaller than those in the more apical cells and are generally fixed as Holmgren canals. 7. In the apical cell of Osmunda, the vacuoles are large and of the same order as those found in the cambium during the growing season. These vacuoles are divided passively into much smaller ones by the series of cell divisions which produce the numerous small cells of the primary meristem. As these cells finally enlarge, the included vacuoles grow and fuse to form the typical large, central vacuole of the differentiated plant cell. Development is here from large, through small, back again to large vacuoles. This is, in general, the course of vacuolar evolution in all of the primary meristems investigated. i 44 Conway Zirkle: 8. In the primary meristem the vacuoles assumed the several forms found by Bailey (2) in the cambium. 9. In no case were vacuoles found to be derived from tonoplasts, hydroleucites, mitochondria, or any other form of cytoplasmic inclusion. They reproduce by the divison of preexisting vacuoles. 10. Vacuoles were never observed to originate de novo. However, in a streaming cell where the vacuoles are constantly fragmenting, changing their shape, and fusing again, neoformation of vacuoles may be obscured. 11. Resemblances between the Golgi appararatus of animal cells and various forms of the plant vacuole have been noted by numerous investigators. A reticulte, tannin-filled vacuole answers the present definition of the Golgi apparatus. Any decision as to whether or not the Golgi apparatus is completely homologous with the plant vacuole must await further investigation of the animal cell. 12. A tannin-free core extends through the root cap in Pinus and Polygonum (Figs. 1, 2). Possible traces of such a core are in the root tips of Robinia and Phaseolus. No such structure is found in the roots of Zea and Osmunda, although the latter has a tannin-filled root cap. The phylogenetic distribution of this core has not been adequately traced. It seems to be quite widespread and is so clear cut, regular, and distinct as to suggest that it performs some specific function. The author wishes to acknowledge his indebtedness to Miss Anna F. Faull, who made the drawings used in the paper. Arnold Arboretum and Bussey Institution, Harvard University. Literature cited. 1. Bailey, I. W.: Significance of the cambium in the study of certain physio- logical problems. J. gen. Physiol. 2, 519—533 (1920). — 2. Bailey, I. W.: The cam- bium and its derivative tissues. V. A reconnaissance of the vacuome in living cells. Z. Zellforsch. 10, 651—682 (1930). — 3. Bensley, R. R.: On the nature of the canalicular apparatus of animal cells. Biol. Bull. Mar. biol. Labor. Wood's Hole 19, 179—194 (1910). — 4. 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Explanation of Plates I, 11, III, IV. The photographs reproduced in Plates 1, 2, 3 were made with a Zeiss camera. Zettnow's solution of copper sulphate and potassium bichromate was used as a color screen. Unless otherwise stated, the material is fixed with a mixture of chromic sulphate and formaldehyde and stained w^ith Heidenhain's hamatoxylin. Plate /. Fig. 1. Longitudinal section of root tip of Pinus Strohus x 36, showing peri- pheral distribution of tannin and colorless core, fixed with a mixture of ferrous sulphate and formaldehyde, and unstained. Fig. 2. Longitudinal section of root tip of Polygonum sachalinense X 90. The black precipitate in certain cells is due to tannin. Fig. 3. Cells from longitudinal section of root tip of Pinus Strohus x 900. The row of cells on the left has 2 kinds of vacuoles, both containing tannin. The vacuoles in the row on the right contain no tannin. Fig. 4. Cells from border of central cylinder of root tip of Pinus Strohus x lOCK), immediately posterior to the "barbs" illustrated in Fig. 1, showing trnasitional region between tannin-filled cells in the left and cells which contain no tannin on the right. Fig. 5. Longitudinal section of root tip of Rohinia Pseudo- Acacia x 70, showing a precipitate in the cortical cells and light region in center of root cap. Plate 11. Fig. 6. Tannin-bearing cells in outer cortex of root tip of Pinus Strohus x 1000. Fig. 7. Elongated tannin-flled vacuoles in anterior portion of root tip of Pinus Strohus X 1000. Fig. 8. Reticular tannin-filled vacuoles in cells near base of embryonic leaf of Pinus Strohus x 2600. Fig. 9. Splitting metaphase chromosomes in tannin-bearing cells in root tip of Pinus Strohus x 2400. Fig. 10. Transitional cells between tannin-filled and tannin-free cells in anterior portion of root tip of Pinus Strohus x 750. Plate III, Fig. 11. Longitudinal section of apex of long shoot of Pinus Strohus X lOCK), showing nuclei, vacuoles, and immature plastids. Fig. 12. Longitudinal section of apex of shoot of Fraxinus americana x 800. Fig. 13. Longitudinal section of apex of short shoot of Pinus Strohus x 975, showing tannin-filled vacuoles in primary meristematic cells. Fig. 14. Longitudinal section through growing point of root tip of Polygonum sachalinense x 1000. The apez of dermatogen, periblem, and plerome are shown. The lightly staining cells at the top belong to the root cap. Fig. 15. Procambial cells in root tip of Polygonum sachalinense x 2400, showing chains of small vacuoles forming Holmgren canals. Fig. 16. Longitudinal section through root tip of Osmunda Claytoniana x 400, showing vacuolate apical cells. Vacuoles in primary Meristems. Plate IV, 47 Fig. 17. Reticulate tannin-filled vacuoles in base of embryonic leaf of Pinus Strohus X 1400. Fig. 18. Same as Fig. 17, x 1400. Fig. 19. Tannin-filled and tannin-outlined vacuoles in cells in root tip of Pinus Strohus X 1400. The black angular rods are tannin-filled vacuoles. Fig. 20. Same as Figs. 17 and 18, X 1400. Fig. 21. Longitudinal section of apical cell of Lunularia vulgaris x 1400, showing Holmgren canals and precipitate. Fig. 22. Optical longitudinal section through growing region in root tip of Rohinia Pseudo-Acacia x 1400, showing vacuoles of various shapes and precipitates. Fig. 23. Procambial cell in root tip of Rohinia Pseudo-Acacia x 1400, showing Holmgren canals. Fig. 24. Longitudinal section through base of embryonic leaf of Pinus Strohus X 1400, showing reticulate vacuoles, some tannin-filled, some partially filled, and some tannin-free. Fig. 25. Longitudinal section through root tip of Zea Mays x 1400, showing vacuoles of calyptrogen (upper cell), dermatogen (second layer), periblem (third layer), and plerome (bottom cell). Fig. 26. Procambial cell in root tip of Zea Mays x 1400, showing Holmgren canals. Fig. 27. Longitudinal section through apical and daughter cells of Lumilaria vulgaris x 1400, showing Holmgren canals and precipitates. Fig. 28. Longitudinal section through apex of shoot of Rohinia Pseudo-Acacia X 1400, showing vacuoles of apical meristem. t , Z. f. Zellforschung u. mikr. Anatomie. Bd. 16. Tafel I. Conway ZiRKLE, Vacuoles in primary Meristems. Verlag von Julius Springer in Berlin INTENTIONAL SECOND EXPOSURE Z. f. Zollforschuno; u. mikr. Anatoniio. Bd. 16. TiiM I. '^ Conway Zirkle, Vacuoles in primary :Moristeins. Vcrlag- von .Iiiliu> Spriiijier in Berlin Z. f. Zellforschung u. mikr. Anatomie. Bd. 16. Tafel II. Conway Zirkle, Vacuoles in primary Meristems. Verlag von Julius Springer in Berlin. Z. f. Zellforschung n. mikr. Anatomie. Bd. 10. Tafol II. * Conway Zirkle, Vacuoles iu primary Meristeius. Verlai? von Julius Spriimer in Berlin. INTENTIONAL SECOND EXPOSURE I Z. f . Zellforschung u. mikr. Anatomie. Bd. 16. Tafel III. Conway Zirkle, Vacuoles in primary Meristems. Verlag von Julius Springer in Berlin. Z. f. Zellforsclnmg u. mikr. Anatomie. Bd. 16. Tcifcl Tir. 1 r :/ i Conway Zikklk, V^acuoles in primary Meristems. Verlag von Julius Sprinf?er in Boi-lin. INTENTIONAL SECOND EXPOSURE Z. f . Zellforschung u. mikr. Anatomie. Bd. 16. Tafel IV. Conway Zirkle, Vacuoles in primary Meristems. Verlag von Julius Springer in Berlin. t Z. f. Zellforschung u. mikr. Anatomie. Bd. 16. Tafel IV. f Conway Zirkle, Vacuoles in primary Meristenis. Verlag von JuliuB Springer in Berlin. INTENTIONAL SECOND EXPOSURE