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Progress Report Nsf Research Grant G13261 The PROGRESS REPORT NSF RESEARCH GRANT G13261 THE BEARING OF CYTOLOGICAL AND CERTAIN PHYSIOLOGICAL DATA ON AN INTERPRETATION OF THE MORPHOLOGY AND TAXONOMY OF THE CHARACEAE 110 tziA k/s3, 7° DEPARTMENT OF BIOLOGY UNIVERSITY OF LOUISVILLE LOUISVILLE 8, KENTUCKY January 1, 1961 Progress Report G 13261 January 1, 1961 PROGRESS REPORT This report contains the data, mostly of a cytological nature, obtained during the first six months of the project period for this study of the Characeae. For convenience, the report has been divided into sections as follows: 1) Project Personnel and Cooperating Botanists 2) General Description of Plant Groups Being Studied, 3) Cytological Methods. 4) Preliminary Account of Chromosome Numbers 5) Polyploidy and GeneraliCytology in the Characeae 6) A Special Study of Nitella flexilis in Progress 7) A Study of Chars braunii Gmelin Principal Investigator Arland Hotchkiss PROGRESS REPORT G-13261 1) Project Personnel and Cooperating Botanists. The efforts of the principal investigator in carrying out the aims and objectives of this study of the Characeae were greatly enhanced and strengthened by the generous contributions of time, labor and guidance expended by quite a long list of individuals. The project was very fortunate in securing the part-time services of a competent cytologist in the person of my wife, who gave invaluable aid without pay, in the laborious task of preparing and analyzing chromosome counts in the home lab and on trips to the field. Mr. Donald Tindall (graduate student) in July, 1960, began research on the Characeae with particular emphasis on the ecology and the complete life history (including cytology) of Nitella flexilis in Doe Run, Kentucky. He aided in collect- ing in Kentucky and Indiana on numerous occasions, and in August 1960 brought back some 20 valuable collections as listed below from a first expedition to northern Mexico. He is planning additional trips to Mexico in 1961. Dr. Richard Wood, University of Rhode Island, contributed to the study from its beginning, and more recently sent the first shipments of many valuable specimens from Australia as noted below. Dr. and Mrs. F. K. Daily (Butler University) guided and fed us in the field, and contributed valuable specimens, and information on collecting in Indiana. Specimens have also been received from the following: Dr. V. Proctor - Texas Tech. Dr. G. Cole - Arizona State College Dr. L. A. Whitford - North Carolina State College Mr. Gene Blankenbaker - Michigan State University 2. Description of the Plant Groups The charophytes are simple aquatic plants, worldwide in distribution, usually in fresh water, agreeing with the green algae in pigmentation, food reserves and cell morphology, but because of the distinctive complexity of organization in their stem nodes and internodes, and whorled branchlets, and especi- ally their elaborate gametangial structures, they are placed in a separate Class Charophyceae of the Chlorophyta by some workers as G. M. Smith, or st 11 further removed from green algae into a separate Division Charophyta by others as Groves and Bullock-Webster, Zaneveld, following Sachs. Within the group of charophytes there is a single Order Charales usually considered to have a single extant family, Characeae, subdivided into two tribes Chareae and Nitelleae. -1- G.-13261 -2- Progress Report There is some evidence which might support raising the two tribes to the status of two or more subfamilies or even families but this has not been done formally. The Tribe Nitelleae contains two genera, one of which, Nitella with 153 species, is the largest in the entire group. The other genus, Tolypella, has but 13 species. The Nitelleae are entirely ecorticate and have two tiers of 5 crown cells sur- mounting the oogonium. In Nitella the branchlets are forked, stem branches two per node, the oospore is laterally com- pressed, the antheridium terminal in a branchlet furcation. In Tolypella the branchlets are monopodial, stem branches more than two per node, the oospore terete in section, the antheri- dium is lateral in position. Important specific characters in Nitella include the kinds of branchlets present, number of furdations in the branchlets, number of cells in the dactyls (ultimate rays), presence of heads, enveloping jelly, spore characters and monoecious or dioecious species. The Tribe Chareae contains four or five genera of which Chara with 116 species is the largest. The others have from one to four species apiece. The Chareae are mostly corticated but a few are entirely ecorticate in the stem and branchlet internodes, as in the Nitelleae. In every case the oogonium is capped by a single tier of 5 crown cells, the brandhlets monopodial and never forked. Important specific characters in the Genus Chara include the degree of cortication, number of whorls of stTFUlodes, arrangement of gametangia on the branchlets, spore characters, monoecious and dioecious species. A Key to the Extant Genera A, Cells of the coronula in two superimposed rows of five cells each .............................................. NITELLEAE 1. Antheridia terminal in the furcations of the branchlets; oogonia lateral; oospores elliptic in transverse section ............................ 1) Nitella 1. Antheridia and oogonia lateral at the branchlet- nodes; oospores terete in transverse section • . 2)To1ype1la AA. Cells of the coronula in one single row of five cells CHAPEAE 2. Stipulodes and bracteoles absent. 3. Bract-cells also absent. Branchlets of 3:4 segments ................................ 3)Protochara 3, Bract-cells 1-2, very long. Branchlets of 2-3 very long segments .................... 4)Nitellopsis 2. Stipulodes always present, sometimes rudi- mentary. Branchlets simple, of L. or more segments. Bract-cells normally L. or more, or rudimentary. Bracteoles also may be rudi- mentary or absent. 4. Oogonia and antheridia produced from separate peripheral cells of the node and situated side by side. Stem corticate ........................ 5) Lychnothamnus 4. Oogonia and antheridia produced from the same peripheral cell of the node. 5. Oogonium normally situated below the antheridium. Stem corticate ........ 6)Lamprothamnium 5. Oogonium situated above the antheridium. Stem corticate or ecorticate 7)Chara Progress Report G 13261 Fossil forms of the Charophytes are well known in paleo- botany and are described and figured in the standard texts by H. N. Andrews, C. A. Arnold, and J. Walton. Because many members of the charophytes secrete a limeshell around the oogonium, this portion of the plant is easily fossilized; other parts of the plant have also been found in the fossil state. Several genera are known from the Devonian, Paleo- chara with six spiral cells is from the Carboniferous, entire plants of the Jurassic genus Clavator were described by Harris from England. Although gyrogonites from the late Triassic up to recent times are well known, little has been done with recent Pleistocene forms which are the immediate ancestors of our present day species. 3) Cytological Methods Fruiting tips of plants are snipped off and either fixed immediately for examination later, or prepared for squashing directly in aceto-ortein. In the latter case, single anther- idia of an appropriate size (age) are teased off the plant branchlets into drops of stain. The slide is warmed slightly and the material is squashed directly under a cover-slip with appropriate pressure. Warming and squashing continue until a proper degree of spread is achieved. The slides are then examined, chromosomes are counted, drawings end photographs are made as required. Slides with good preparations are made permanent by trans- ferring to 90% alcohol for a few hours and then in 100% al- cohol for at least two hours. The coverslip is allowed to float off the slide inverted in alcohol or it may be carefully pried off with a razor blade. Mounting in Euparal is done directly from the absolute alcohol. Field collections which were fixed in either 3:1 (100 parts absolute alcohol: 33 parts glacial acetic acid) or Carnoy's (100 parts absolute alcohol: 16 parts glacial acetic acid; 50 parts chloroform) for at least 12 hours, then transferred to 70% alcohol and kept preferably refrigerated (at least cool) have proved eminently satisfactory provided the fixative was made up fresh for each collection. Australian and Mexican materials and controls from all collecting in 1960 have been prepared in this way and show perfect preservation when later transferred to aceto,-orcein stain for approximately an hour before squashing in the stain on a slide. 4) In the following tables, are shown our own counts of chromo- some numbers for the year 1960 and also, for comparison, the preceding years back to 1957. The perfect agreement with ourselves over the years merely reflects our practice of eliminating from the summary those results we later believed to be based on mistakes inPoounting or in identification of material. No attempt has been made here to make comparisons with numbers in the literature except our own. Other collec- tors names and people who aided in our own collections are noted. In several instances in Tables 2 and 3, complete identifi- cations of the specimens collected by Tindall and Wood have , -14- Progress Reiport G 13261 not yet been made. Table 1. Chromosome counts in the Characeae 1957-1960 Species Chromosome No. Location Dates Chareae 1. Chara aspera Deth. N 14 Lake Wawasee, Ind. 1960 with Tindall ex. N. Wild. 2. Chars braunii Gmelin 14 Near Lake Gauvreau, 1960 Quebec 14 Masson, Quebec 1957,58,59,60 14 Black Rapids, Ontario 1957,59,60 14 Picton, Ont. 1960 14 Keuka Lake, N.Y. 1959 14 Mecklenburg,N.Y. 1960 14 Cayuta Lake, N.Y. 1959,60 14 Lake Waramaug,Conm. 1958: Wood var. schweinitzii 114 Starve Hollow 1960 with Tindall Hatchery,Ind. 3. Chara brittonii 114 Cabin Creek Bog,Ind. 1960 Daily T.F. Allen ex Rob. 4. Chara contraria 28 Cayutaville, N.Y. 1960 Br. ex Kutz 28 Lansing ,Michigan 1960 Blankenbaker 28 Lake Wawasee, Ind.
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