AN mvzsmmw 0% ma mwaow 0F mummy-:23 'EATONH, 5; snow: 50mg CAS'E‘ANEA; Am: 5: IOMENTQSA I Thurs-ts fas- Hm Dun-ca of M. 3. , ' MECHLQIKN STATE UNEVERSITY - Julie Hitt 195-6 no-O-I LIBRARY Michigan State University THESIS ABSTRACT AN INVESTIGATION OF THE TAXONOMY OF CHEILANTHES EATONII, Q, EATONII FORMA CASTANEA, AND E, TOMENTOSA by Julie Hitt Three xerophytic ferns, Cheilanthes eatonii, g, eatonii f. castanea, and Q, tomentosa have many similar characteris— tics and appear to be closely related. These three taxa were studied primarily by morphological comparison in an attempt to determine their proper taxonomic rank. Spores from living plants were sterilized and sown both on a mineral salt agar medium and soil for gametophytic study. Spores from living plants and herbarium Specimens were aceto- lyzed for detailed study. Measurements of the following characters were recorded: 1) Spore diameter in the equatorial plane 2) length of the blade and stipe 5) width of the blade 4) length and width of the cells of the upper and lower leaf epidermal surfaces 5) length and width of the stomates 6) length and width of the rhizome, stipe, rachis, and pinna midrib scales Julie Hitt The indusium and sporangium of each taxon were generally compared. Cheilanthes tomentosa is distinguished from the other two taxa by these characters: 1) longer and narrower stipe, rachis, and pinna midrib scales 2) longer and broader fronds 3) greater blade-length:stipe-length ratio The scales of Q, eatonii and g, eatonii f. castanea have a broad base and gradually taper toward the tip with those of “Q. eatonii being broader at the base than those of Q, eatonii f. castanea. Their frond measurements and blade-length:stipe- length ratio are similar. All three taxa were found to be triploid apogamous hybrids. Based on constant morphological characters, 9, tomentosa and g, eatonii may be properly classified as species. Cheilanthes eatonii f. castanea is classified as a forma be- cause of its close alliance to Q, eatonii in morphological characters. These conclusions are also supported by cyto- logical evidence. AN INVESTIGATION OF THE TAXONOMY OF CHEILANTHES EATONII, Q, EATONII FORMA CASTANEA, AND Q. TOMENTOSA BY Julie Hitt A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of BOtany and Plant Pathology 1966 f/ --/’-\ N. . , // I v . ' . . . ”I,” /‘ V "' ACKNOWLEDGMENT S To Dr. Irving W. Knobloch, I express my sincere appreci- ation for his inspiration, continued guidance.and help, and above all, his patience, which have made this study possible. Drs. William B. Drew, Gerald W. Prescott, and John H. Beaman, members of my guidance committee, have been most helpful in their suggestions and constructive criticisms of this work. I wish to thank Dr. Donovan S. Correll of the Texas Research Foundation and Mr. David B. Lellinger of the United States National Herbarium who have loaned me herbarium speci- mens and supplied additional information. Special thanks is given to Lenette Atkinson (Mrs. Goeffroy) for her valuable information concerning the gameto— phytes of this study. Dr. C. V. Morton's English translation of the original description of Cheilanthes tomentosa is greatly appreciated. Dr. Aureal T. Cross is to be thanked for his help in spore techniques. I should also like to acknowledge Dr. Warren H. Wagner, Jr., the University of Michigan, for his helpful suggestions and information throughout this study. The encouragement, suggestions, and criticisms of Mr. Dennis C. Jackson and Mr. Ronald W. Hoham throughout this study are deeply appreciated. I should like to especially thank Miss Janice M. Glime for her continued inspiration, suggestions, and tireless efforts throughout this study, particularly in the preliminary writing and preparation of this paper. ii TABLE OF CONTENTS Page INTRODUCTION . . . . . . . . . . . . . . . . . . . . 1 OCCURRENCE AND HABITAT . . . . . . . . . . . . . . . 7 COMPARATIVE MORPHOLOGY . . . . . . . . . . . . . . . 7 MATERIALS AND METHODS . . . . . . . . . . . . . 7 RESULTS AND DISCUSSION . . . . . . . . . . . . 10 SPORES . . . . . . . . . . . . . . . . . . 10 GAMETOPHYTE . . . . . . . . . . . . . . . 18 APOGAMY. . . . . . . . . . . . . . . . . . 21 MATURE SPOROPHYTE. . . . . . . . . . . . . 25 RHIZOME . . . . . . . . . . . . . . . 25 FROND . . . . . . . . . . . . . . . . 27 STIPE AND RACHIS . . . . . . . . 27 BLADE. . . . . . . . . . . . . . 28 EPIDERMAL CELLS . . . . . . . . . . . 31 HAIRS AND SCALES . . . . . . . . . . 35 INDUSIUM AND SPORANGIUM . . . . . . . 58 SUMMARY AND CONCLUSIONS. . . . . . . . . . . . . . . 40 PLATES . . . . . . . . . . . . . . . . . . . . . . . 57 BIBLIOGRAPHY . . . . . . . . . . . . . . . . . . . . 78 APPENDIX . . . . . . . . . . . . . . . . . . . . . . 82 iii LIST OF TABLES TABLE Page I. Spore Diameter . . . . . . . . . . . . . . . . 14 II. Rhizome Scale Measurements . . . . . . . . . . 26 III. Frond Measurements . . . . . . . . . . . . . . 50 IV. Measurements of Epidermal Cells and Stomata. . 54 V. Larger Scale Measurements. . . . . . . . . . . 36 APPENDIX TABLES I. Spore Diameter . . . . . . . . . . . . . . . . 87 II. Rhizome Scale Measurements . . . . . . . . . . 88 III. Frond Measurements . . . . . . . . . . . . . . 85 IV. Measurements of Epidermal Cells and Stomata. . 89 V. Larger Scale Measurements. . . . . . . . . . . 89 iv FIGURE LIST OF FIGURES . Cheilanthes tomentosa spore diameter . Cheilanthes eatonii spore diameter . . Cheilanthes eatonii f. castanea spore diameter . . . . . . . . . . . . . . . Composite polygonal graph. . . . . . . Cheilanthes eatonii polygonal graph. . Cheilanthes eatonii f. castanea polygonal graph. . . . . . . . . . . . . . . . . Cheilanthes tomentosa polygonal graph. Page 15 16 17 45 47 49 51 INTRODUCTION The genus Cheilanthes was originally described by Swartz in 1806 and includes 150-200 Species. The systematic posi- tion of this genus seems to be in a great state of confusion at present and the boundary of the genus varies according to the authority. This is evident when the position of the genus is compared in various systems of classification. Cheilanthes is usually placed in the Gymnogrammoid fern group of the family Polypodiaceae as designated by Bower (Vol. 3, 1928). The kinship of this group "is based partly on positive characters of external form and internal anatomy: partly upon the rather negative soral characters, and particularly upon the absence of a true indusium, with its corollary, an indefinite soral construction.. They may be held to represent a plexus 2£_phyletic lines all traceable back with probability to Ferns with marginal sporangia of larger size than theirs such as the present-day Osmundaceae and Schizaeaceae." More specifically Bower is referring to the advanced members Anemia and Mohria of the Schizaeaceae. The sporangia and vascular systems of the latter two genera are similar to those of Cheilanthes, and Mohria has dermal scales which are found in Cheilanthes and most other Gymnogrammoids. More specifi- cally Cheilanthes is placed in the fourth seetion of this group, which includes specialized xerophytic genera, such as_Pellaea, Doryopteris, Cheilanthes, Notholaena, Saffordia, and Trachypteris. Bower (Vol, 5, 1928) makes it explicit, that the derivation of the Cheilanthoid Ferns from the living Schizaeaceae is not clear-cut, but "the variety and cogency of the comparisons may be held as indicating a reasonable probability of its truth." Copeland (1947) states that Cheilanthes has long been a "puzzling genus" and places it in the family Pteridaceae. The ancestors of this family may be found in the Schizaeaceae and may involve more than a single line of descent. Concern- ing Cheilanthes Copeland (1947) contends that the Schizaeaceae ancestry should be an alternate choice with a Dennstaedtioid origin as the other choice. The latter choice would be based on soral affinity found in Hypolepis, Pteris, and Cheilanthes-- reflexed margins protecting the sori in all cases. In other words, Cheilanthes was derived from an indusiate ancestor according to this theory. Hypolepis is related to Dennstaedtia with a hiSpid stipe being common in primitive members of both genera. However, he does recognize that the generic affinities in the group are not well-defined. Pichi-Sermolli (1957-1960) agrees with Copeland in the possible derivation of Cheilanthes from the Schizaeaceae. He states that "the Gymnogrammoid, Cheilanthoid, Adiantoid and Vittarioid ferns may be held to represent phyletic lines which branched off early from a common stock." Holttum (1947) supports the possible Schizaeoid ancestry of Cheilanthes. He states: "Several authors have pointed out the resemblance of Cheilanthes and its allies to Mohria, a genus of the Schizaeaceae. Mohria is advanced, as compared with other Schizaeaceae, in having scales instead of hairs, and in its vascular structure. It has single Sporangia at its vein-endings, with the margin of the lamina reflexed to cover them as in Cheilanthes. A remarkable primitive ally of Cheilanthes, the genus Singpteris, also has single terminal sporangia; this may well be an indication of Schizaeoid affinity." Holttum (1947) places Cheilanthes in the subfamily Gymnogrammoideae in the family Adiantaceae, which also in- cludes the subfamily Vittarioideae. He does not support a Pteroid derivation for the Cheilanthoid group, one of his main contentions being the difference in rachis shape. In the Pteroid group "the midribs of the leaflets are deeply grooved with raised edges, these edges being decurrent upon the edges of the groove of the rachis, which is interrupted to receive them." Such a rachis shape does not exist in the Cheilanthoid group. Christensen (1958) places Cheilanthes in the family Poly- podiaceae, subfamily Gymnogrammeoideae. He says the 27 genera in this subfamily "are of uncertain mutual relationship and probably representing more evolutionary lines, but most of them no doubt related to the Pteridoideae, with which they were associated by most authors." Cheilanthes is placed in the tribe Cheilantheae about which he says: "The 8 genera of this tribe (Adiantopsis, Cheilanthes, Notholaena, Sinop- teris, Pellaeathoryopteris, Saffordia, and Cheilanthopsis) run confusingly together and the classification of the species is quite uncertain." Diels (1902) also places Cheilanthes in a Pteroid series in Engler and Prantlwsclassification. In addition, Ching (1940) places Cheilanthes in the family Sinopteridaceae, a Pteroidn-Gymnogrammeoid series derivative. This series in- cludes the families Hypolepidaceae, Pteridaceae, Sinopterida— ceae, Gymnogrammaceae, Adiantaceae, and Ceratopteridaceae in a direct linear order of derivation. In addition to the question of the ancestry and classifi- cation of Cheilanthes, the specific as well as the generic affinities of the Cheilanthoid group are not well defined as Christensen (1958) and Copeland (1947), among others, have mentioned. The purpose of this research is to study several taxa of Cheilanthes which appear to be closely related in an attempt to determine the correct level of taxonomic classification. Cheilanthes tomentosa Link and g, eatonii Baker, ex Hook.& Bak. from investigation appear to be two separate Species, but Q, eatonii Baker f. castanea (Maxon) Correll is a problematical taxon which has been classified both as a species and as a form of g, eatonii. As is evident from an examination of the original descriptions below, these three taxa have many characteristics in common and are separated only by minute differences. Cheilanthes tomentosa Link (Link, 1855--English trans- lation by C. V. Morton) Frond tripinnatifid, the stipes tomentose and scaly- pilose, the pinnae thinly tomentose above and densely tomentose beneath, the crenations minute. We accept the name Cheilanthes tomentosa for a fern Specimen grown from seeds (spores) from Mexico. Fronds 6-8 inches long; stipes and rachis pale green, with a thin tomentum, the hairs flattened and thus scalelike; pinnae scarcely an inch long, triangular in outline, the pinnules crenate-pinnatifid, the crenations minute a line (i.e. 2 mm.) in diameter, tomentose above where soon the green color shines through, very densely tomentose beneath. Indusia greenish, glabrous. Cheilanthes Eatonii, Baker (Hooker and Baker, 1874) Stipe tufted, 5-6 inches long, wiry, erect, densely clothed with pale-brown linear subulate scales; frond 5-8 inches long, 1i—2 inches broad, ovate-lanceolate, tripinnatifid; lower pinnae distant, alternate or Opposite, deltoid; pinnules linear-oblong, pinnatifid; rachis rigid, covered with scales like the stipe which also cover thickly the midrib of the pinnae beneath; texture coriaceous: upper surface densely clothed with white woolly tomentum, lower also densely matted, the margin of the segments in- curved.--§, tomentosa, Hk. Sp. 2. p. 96. (in part), t. 109. A. non Link. Habitat. Gathered in an expedition from Western Texas to El Paso, New Mexico, October 1849, C. Wright, No. 816.--This differs from g, tomentosa by being coated with distinct scales instead of mere woolly hairs on the stipe, rachis, and midrib of the pinnae beneath, and by being matted with tomentum on the upper surface. Cheilanthes castanea Maxon, Sp. nov. (Maxon, 1919) Rhizome short-creeping, nodose or short-branching, the divisions 1 to 5 cm. long, less than 1 cm. thick, densely paleaceous, the scales oblique, imbricate, falcate, 5 to 5.5 mm. long, about 0.6 mm. broad at the base, subulate attenuate, rather lax, tawny, with a dis- tinct glossy, dark brown, sclerotic, median stripe extend- ing nearly to the filiform, flexuous tip. Fronds, few, very closely distichous, erect, 16 to 50 cm. long; stipe 9 u>18 cm. long, castaneous, sublustrous beneath a thin covering of appressed to rigidly ascending, pale tawny scales, the larger of these linear-attenuate, underlaid by minute acicular ones; lamina linear to linear-oblong, long-acuminate, 7 to 17 cm. long, 2 to 4 cm. broad, tripinnate, the rachis similar to the stipe but with larger and more numerous scales; larger pinnae 8 to 12 pairs, spreading (or with age oblique and involute), distant, sesSile, deltoid-oblong, acutish, slightly in- equilateral; secondary rachises persistently paleaceous, the scales rather large, flaccid, imbricate, linear- deltoid to ovate, long-acuminate, firmly attached at the cordate base, erose-denticulate, tawny; segments of the larger pinnules mostly 5 or 5, oblong, entire, unequal, the terminal ones the longest (up to 4 mm. long), with a cuneate base; segments loosely but copiously tomentose beneath with Spirally crispate, light castaneous hairs, glabrate above, the few similar but griseouS hairs easily deciduous; segments mostly fertile, the recurved margin gradually thinner, Slightly repand, minutely sinuate, pale, hardly forming a proper indusium; sporangia not concealed at maturity, the tomentum separating evenly from the indusiiform margin. Leaf tissue rather rigidly herbaceous, dull grayish green. Cheilanthes Eatonii Baker forma castanea (Maxon) Correll, 1949 Comb. nov. Cheilanthes castanea Maxon When extremes of typical 9, Eatonii and forma castanea are found they are so distinctive that one would immediately consider them to be specifically different. Unfortunately, very little material of extreme f. castanea exists in herbaria. An overwhelming amount of material does exist, however, which grades into one or the other of these two forms. It is simply an arbitrary matter as to which category they should be relegated. I have placed all plants which have all or part of their fronds tending to be glabrescent on the upper surface of their segments into f. castanea. I have designated those plants with segments hoary and densely tomentose above as the typical form. Habitat same as that of the typical form. OCCURRENCE AND HABITAT The distribution of these taxa is almost the same accord- ing to several authors (Wherry, 1961; Fernald, 1950; Morton, 1951; Gleason, 1965; Maxon, 1919) and as indicated by locality data on herbarium specimens. Q, tomentosa is found in Mexico, New Mexico, Arizona, Texas, Oklahoma, Missouri, Arkansas, Kansas, Alabama, and Georgia and extends northward to the mountains of western Virginia, North and South Carolina, West Virginia, Tennessee, and Kentucky. This Species has the widest distribution of the taxa included in the study, the ranges of the other two being included within this distribution. g, eatonii is found in Mexico, New Mexico, Arizona, Colorado, Utah, Oklahoma, and Texas. Cheilanthes eatonii f. castanea has approximately the same range as g, eatonii except for Colorado and Utah. These taxa are found primarily in dry areas on wooded hillsides, crevices of rocky Slopes and ledges, or often in Chaparral. They have been reported as high as 8000 feet in altitude (Morton, 1951). COMPARATIVE MORPHOLOGY MATERIALS AND METHODS For this study approximately 150 specimens from the Texas Research Foundation (LL), 60 Specimens from the Smithsonian Institution (US), and 50 specimens from the Michigan State University Herbarium (MSC) were examined. In addition, living plants of Q, eatonii 65e49B, Q, eatonii f. castanea 65-28, 65-50, 65-49D, 64-21, Q, tomentosa 2048, 65-60, and 64-16 were used as sources of Spores and epidermal surfaces of the leaves. Voucher Specimens of these plants have been deposited in the Michigan State University Herbarium. All specimens are cited in the appendix. The following characters were recorded: 1. Spores a. Diameter 2. Gametophyte a. General description b. Gametangia 5. Rhizome a. Scales--base and tip 4. Frond a. Stipe 1) length (comparison to blade also) 2) terete or sulcate 5) color 5. Blade a. Length and width b. Shape c. Descriptions of pinnae, pinnules, and ultimate segments d. Cells of the lower and upper leaf epidermides 6. Hairs and scales a. Stipe b. Rachis c. Pinna midrib 7. Indusium and sporangium a. General description The methods used in studying the individual characters are included at the beginning of each section under the results and discussion. RESULTS AND DISCUSSION SPORES Fronds containing mature sporangia were collected from living material in the greenhouse. These were placed in a 5% household chlorox solution containing a few drops of Tween 20 (wetting agent) and brushed on both sides for 5-10 seconds with a camel‘s hair brush. They were then rised in water and placed in a plant press between blotters for several days prior to sowing for growth studies. Spores to be studied morphologically were collected from herbarium Specimens with forceps and placed in a few cc. of glacial acetic acid. Spore preparations were then made following Erdtman's acetolysis technique (1952). The spores were placed directly into centrifuge tubes containing 5 cc. of acetolysis solution. This solution was prepared by adding drop by drop 1 part concentrated sulfuric acid to 9 parts acetic anhydride (chemically pure). A glass stirring rod was placed in each tube and the tubes were heated for several minutes in a water bath not exceeding 800C. Once this temperature was reached, the heating was stopped and each tube was stirred before centrifuging for several minutes. After centrifuging, the acetolysis solution was decanted and the spores were rinsed once with glacial acetic acid and once with water, centrifuging 10 11 and decanting after each rinse. Since the spores were a very dark brown, they were bleached for easier observation. This was accomplished by pouring a few cc. of household chlorox and leaving until Spores became a light brown. Water was added and the spores were then centrifuged, decanted, and rinsed again with water. The Spores were then stained with a 1% safranine in 50% alcohol solution and rinsed in water. The spores were suspended in a 1:1 glycerine and water prep- aration for at least 10 minutes or could be left for a day or so without affecting the Spores. When slide preparations were to be made, the spore suSpensions were centrifuged and decanted. A drOp of the spore residue was placed on a slide on a warming plate. A coverslip was applied and the Slide was sealed with Canada balsam or clear fingernail polish. The diameter of the spores was measured as the greatest distance in the equatorial plane excluding the perispore. Fifty Spores from each of five specimens of each taxon were measured. The spores were drawn in the same plane with the aid of a camera lucida. To determine the number of Spores per sporangium a single sporangium which had not dehisced was mounted on a slide in a drop of Hoyer's solution. The Spores of all three taxa appear very Similar and no distinguishing characteristics could be observed. The Spores are trilete and globose with crassimarginate smooth lasurae (Plate I, Figs. 7-9). They range in color from light tan to 12 dark brownish—black, the lighter color usually indicating younger Spores. Nayar (1965) reports that members of Cheilanthes have an exine which is composed of an inner and outer layer, the endo- and ecto—exine. In the three taxa of this study this demarcation can not be clearly observed. Although there seem to be two distinct layers in some Spores, it is not a constant character. The exine is smooth or nearly so in all Spores and ranges from 1.5 to 5.5u in thickness. The outline of the protoplast is easily observed. Another layer is present outside the exine which is called the perispore or perine. The exact concept of a periSpore has long been a point of confusion. Bower (1965) states: The plasmodium intrudes between the separating spore-mother-cells forming a rich nutritive medium, which is absorbed in the more primitive ferns in the developing spores; but in certain advanced types it may in part remain as a deposit on the outside of the wall, and is called the perispore (fig. 559). The wall of the Spore itself is often marked by characteristic sculptur- ing which at times gives a basis for systematic com- parison; but in this the perispore is more important. Ferns may in fact be divided into two groups according to the presence or absence of a perispore. None is seen in the Eusporangiate Ferns, nor in the Osmundaceae, Schizaeaceae, Hymenophyllaceae, Cyatheaceae, Davallieae, or in Ceratopteris. In fact, it is absent from all the more primitive ferns, and of the remaining Leptosporbx angiates it is wanting in the Vittarieae, Gymnogrammeae, Polypodieae, and Pterideae; but it is present in the Asplenieae and Aspidieae. The perispore thus possesses a certain value for comparison; but confidence in it as a safe criterion is shaken by the fact that while it is present in Blechnum and Woodwardia it is absent in the closely related Brainea and Doodia. It is clearly a feature adopted late in descent, and restricted to cer- tain circles of affinity. 15 However, Harris (1955) goes on to say, after quoting Bower: It is a common observation that whereas the mature spores of a given species may exhibit a characteristic ornamentation, Spores which are not fully mature lack this ornamentation. Between the smooth immature spores and the fully coated mature spores others may be ob- served which Show a partial accretion of the ornamental layer (Fig. 2g and h). If this arises by the gelation of plasmodial substance on the exterior of the spore, it is a perispore as defined by M011 (1954) and other writers. It is possible, then, that the occurrence of a perispore is more general than supposed by Bower. In some ferns it is difficult to determine whether or - not a distinct perispore is present. Therefore, the terms "sculptive" or "winged" spore have been adopted by workers when referring to a layer outside the exine of doubtful origin (Erdtman, 1952, and Harris, 1955). In the three taxa of this study the periSpore is well-defined and is usually slightly darker in color than the underlying spore. It is smooth and is approximately 1.5-5.5u in thickness. The peri- spore is usually slightly thicker than the exine, but some- times both appear to be about the same in thickness. The diametric measurements of the Spores did not prove to be valuable as a distinguishing characteristic (Table I). Graphs of the measurements Show Q, eatonii f. castanea and .Q. tomentosa to have a nearly normal curve (Figures 1 and 5) while Q, eatonii (Figure 2) tends toward a normal curve with a slight dip near the summit, indicating greater phenotypic variation than the other two taxa. It is to be noted that the measurement range of Q, eatonii f. castanea falls within that of Q, eatonii. The sporangia of each taxon contain 52 spores per sporangium. 14 The spores are uniform in appear- ance and none of them is aborted. Table I. Spore Diameter * Collection Diameter (u) Taxa Collector Number Mean Range .Q. tomentosa I.W.Knobloch 1955 57.8 (48.0-70.4) E.Castetter 65-60 58.9 (44.8-67.2) I.W.Knobloch 847 58.6 (51.2-70.4) I.W.Knobloch 2048 61.1 (48.0-70.4) D.Moore 64-16 61.5 (54.4-75.6) 57.6=Mean of the means .Q. eatonii R.M.Stewart 65-49B 59.1 (51.2-75.6) R.M.Stewart 1112 55.6 (41.6-67.2) I.W.Knobloch 752 51.9 (58.4-70.2) D.S.Correll 25291 51.8 (41.6-60.8) D.S.Correll & 21625 58.5 (41.6-75.6) I.M.Johnston 55.5=Mean of the means ‘Q. eatonii f. B.H.Warnock 65-28 57.4 (51.2-64.0) castanea G.Goodman 64-21 56.2 (51.2-64.0) Lefebure 1161 61.2 (48.0-70.4) J.K.Baker 65-49D 56.5 (48.0-64.0) B.H.Warnock 65-50 59.9 (51.2-67.2) 58.2=Mean of the means 9(- Specimen citations in appendix. 15 .umuOEmap muomm mmoucoEOu monucmHHmcu imp FL P -_,L P ion _ _ _ _ _ 13 E _ — :8 _ _ FF. 3 1mm r r8? 1m.>m 1% _ .fi musmflm 1m¢ __HP-L- mmmum>¢ Two ”HON Ilmh .uom sexods JO Jeqmnn 16 105 F _ .umumEmaU muomm HHSOumm monucmHfiwso 1mm :08 1mm :om L_r_r____p___p_r_____ 1m.mm n mmmnm>4 .N musmflm selods go Jeqmnu 17 .HmuOEMAU muomm mwsmummo .m Hflcoumm mmzucmHflmnu .m mnsmfim imp doe 1mm 108 1mm 10m :ma _ c 105 1mm __e___F__c__FbLS_P___T________ e._ ___ ___ lom 1mm 1N.mm n mmwum>¢ o sszods go Jeqmnu 18 Gametophyte In early January, 1965, spores for sowing were collected from living material and treated in the same manner described in the Spore discussion (p. 10). While the fronds were drying, potting soil was placed in flower pots and autoclaved for one hour under 15 pounds of steam pressure. After the fronds had dried, and the Sporangia had dehisced, the spores were col- lected and approximately half of them sprinkled on the soil. Petri dishes served as covers for the flower pots and these were sealed to the pots with masking tape. The pots were placed under 24-hour fluorescent lighting in trays containing approximately an inch of water. At the same time a nutrient agar solution was prepared using the following formula (Steeves, gt 1., 1955): Knudson's solution of mineral salts Water (glass redistilled) 100 cc. Ca(N03)g-4H20 500 mg. MgSO4-7H20 125 mg. K2HPO4 125 mg . To the above were added 1 cc. of ferric citrate, 1 cc. of trace elements, 50 g. of sucrose, and 1% agar. The solu- tion was heated to melt the agar and poured into 125 ml. Erlenmeyer flasks to a depth of about 5/4 inch. The flasks were stoppered and autoclaved for 20 minutes under 15 pounds of steam pressure. Spores were then sprinkled on the agar medium and the flasks were placed under fluorescent lights. 19 By the end of January most of the agar cultures were contaminated and of no value. At this time Whittier's (1964) method of Spore sterilization was tried. Remaining spores from fronds collected earlier were wet with a 0.1% solution of Tween 80 and placed in a 15% solution of household chlorox for 2 minutes. The spores were collected on filter paper and washed several times with sterile water. The filter papers were placed in envelopes and put between blotters for drying. After drying, the spores were scraped from the filter paper with a sterile knife and placed in a vial containing several cc. of sterile water. The vial contents were transferred aseptically to Knudson's agar medium. The vial contents were transferred aseptically to Knudson's agar medium. One advan- tage of this method is that it allows for a more even distri- bution of the spores over the surface medium. At this time no spores of Q, eatonii 65-49B were available for sowing and so gametophytes of this plant were not available for study. By the end of February prothalli were evident on soil and agar cultures. AS the agar dried in the cultures, the pro- thalli were transferred aseptically to new culture media by means of a transferring needle. By the end of May young sporophytes were evident and some were preserved in FAA. By the middle of June, sporophytes were evident on the soil cultures. The gametophytes of Q, eatonii f. castanea and Q, tgmgnf tosa were examined, but no material of Q, eatonii was 20 available as mentioned previously; hence, complete gameto- phytic comparisons could not be made. Lenette Atkinson (pers. comm.) also supplied additional information and sup- port to the investigator's findings. The gametophyte of Q, tomentosa is usually an elongated plate of cells at first which then broadens into a basically cordate-shaped prothallus with a very broad sinus (Plate II, Figure 1). The margins are slightly irregularly lobed. In many cases, one or both of the thallus wings send out ad- ditional filamentous thalli (Plate II, Figure 5), the result- ing gametophyte having a bizarre appearance. On the agar cultures the gametophytes grew in clumps with those on the upper surface being of the cordate nature and filamentous ones growing into the agar below (Plate II, Figure 2). The thalli are one cell in thickness except in the central area below the sinus where a cushion of cells several layers thick forms. The gametophyte of Q, eatonii f. castanea is similar to that of the previous taxon except that the margins are more deeply lobed, thus giving the entire thallus a much more irregular appearance than that of Q, tomentosa (Plate II, Figure 4). The thalli are often very wavy and somewhat urn- shaped making them difficult to flatten for observation. The filamentous growths of the thallus wings and lower sur- face may again be noted (Plate II, Figures 5 and 6). 21 Atkinson (pers. comm.) observed the thalli of Q, eatonii to be large cornucopia-shaped structures when grown in fibre- sand. In Q, tomentosa, Q, eatonii f. castanea, and Q, eatonii (Atkinson, pers. comm.) antheridia are found near the basal end of the prothallus away from the apical notch. These are subspherical and sessile on the thallus or on a one-celled stalk (Plate II, Figure 2; Plate II, Figure 5). No arche- gonia were observed on any thalli. Unicellular rhizoids occur on the underneath surface along the midrib cushion, but no hairs are observed on any of the thalli. The usefulness of the gametophyte in taxonomic studies is rather doubtful as Wagner (1952) points out: "AS has been shown widely in the literature, and as is probably true of all cordate gametophytes of leptosporangiate ferns, the form is exceedingly plastic, varying in accordance with lighting, temperature, nutrition, crowding, and other external condi- tions." Others, however, find the gametophyte stage of value in their studies. Apogamy All three taxa are apogamous. This is indicated by several observations which will be discussed in detail later: 1) a mound or cushion of cells forms on the lower surface below the sinus on the gametophyte which seems to be a pre- requisite for apogamy, 2) each Sporangium contains 52 22 well-formed spores instead of the usual 64 well-formed spores found in sexual Species, and 5) no archegonia have been observed on the gametophytes. Apogamy in ferns may be defined as the production of a sporophyte, the vascular plant of the asexual generation, from the gametophyte, the nonévascular plant of the sexual generation without the occurrence of syngamy (Whittier and Steeves, 1960). There are basically two types of apogamy: obligate and induced. Obligate apogamy refers to the formation of Sporophytes from the gametophytic tissue as a necessity because fertilization cannot take place. This necessity results from either the antheridia or the arche- gonia or both being absent from the gametophyte. Induced or facultative apogamy refers to the formation of sporophytes from gametophytic tissue by controlling the cultural condi— tions. Under normal conditions these plants would have functional archegonia and antheridia and fertilization could take place to sexually produce the sporophyte. Whittier (1965) describes the development of sporphytes apogamously in Q, tomentosa as follows: "After the cordate prothallus thickens, a small tan area appears in the cushion region behind the apical notch. This color results from a modification of chloroplasts in the meristematic cells of this region. The area increases in size and becomes brown. A mound of dark-green cells with sporophytic hairs is pro- duced from cells of the brown area. From this mound the leaf 25 appears first, with the root following shortly thereafter.“ This mound of darkened cells was observed also in the gametophytes investigated in this study. Whittier (1962) and other workers have concluded that a thickened thallial structure must be formed before apogamy can occur. In apogamy the central cytological problem is that of reconciling the absence of a sexual nuclear fusion with the presence of an apparently normal meiotic process in the development of spores in the sporophyte (Manton, 1950). There must be some compensating process to stabilize the number of chromosomes. Early work on this idea was done by Allen (1914) with Agpidium falcatum,$w. (now Cyrtomium falcatum Presl). He correctly stated that the meiotic process produced Spores with the same chromosOme number as the parent and that the sporophyte and gametophyte were the same in nuclear content and of the same chromosome number as the parent. He found Sporangia with eight spore mother cells, sixteen Spore mother cells, and some intermediates. In 1919 Steil studied intermediate nuclear stages as found in Nephrodium hirtipes Hk. [now Dryopteris atrata (Wall.) Ching]. These stages were the result of incomplete nuclear division im- mediately preceding meiosis causing the nuclear content of the Spore mother cell to be doubled momentarily. This was the key to the compensation mechanism. 24 Manton (1950) describes the cytological processes taking place in the sporangia of the apogomously-produced sporophyte. The early stages of sporangial development are identical in a sexual or apogamous sporophyte. A very exact set of cell cleavages occurs in what was originally a single superficial cell. A short filament develops, the terminal cell of which undergoes four oblique cleavages. By this means the sporangium wall is separated from the central tetrahedral cell. This central cell next undergoes a set of cleavages which separates the nutritive tapetal layer from the archeSporium. The archeSporium then undergoes four equal mitoses resulting in a central unit of sixteen cells. These cells enlarge to become spore mother cells which undergo meiosis each giving rise to four spores. In apogamous ferns the sporongial development is the same until the onset of mitoses in the archeSporium. At this stage there are four possibilities for further development: (1) All four archesporial cleavages may take place and sixteen spore mother cells formed as in the sexual Species. (2) The first three cleavages may be perfect, but the fourth is imperfect. Metaphase starts in the eight arche- sporial cells as usual with the chromatids attached to the Spindle. At anaphase there is no chromatid movement to the poles and no cleavage of cytoplasm. The nucleus becomes a restitution one and eventually becomes a spore mother cell with meiosis occurring and resulting in only 52 spores with the 2g_number of chromosomes. 25 (5) A variation of (2) may occur in which the nuclei become irregularly lobed and have partial cross walls result- ing in unequal constricted portions. Because the distribution of chromosomes to the constricted portions is random, there is a good chance that the nuclei will be genetically un- balanced and the spores will probably abort. (4) Two cleavages of the archeSporium may be affected re— sulting in a four-celled state with four large mother cells at meiosis each with four times the normal number of chromo- somes. The pairing of chromosomes is restricted to sister chromosomes and the ripe sporangia yield sixteen large spores with twice the chromosome number of the parent. This may have some connection with polyploidy. The three taxa in this study all have 52 good spores per Sporangium, as was previously mentioned, and therefore seem to fit in the second developmental possibility. Mature SporOphyte Rhizome The rhizome scales were mounted on a slide in Hoyer's solution for study. The length was measured from the base to the tip of the scale and the width was measured at the widest point near the base of the scale. Measurements were made using scales from both the base and the tip of the rhizome. The rhizome is very similar in all three taxa in being stout with fronds arising in very close proximity to one another. 26 It is densely covered with imbricate scales which are linear to linear-lanceolate and tawny-colored. Those at the base usually have a central dark brownish-black stripe which may vary in occupying the central third of the scale up to a short distance from the margin. Scales at the tip may be uniformly tawny or have a small central stripe (Plate I, Figures 1-6). Measurements of the scales show only slight differences among the three taxa (Table II). Q, tomentosa has longer, narrower scales which are characteristic of its scales on other parts of the plant to be discussed later. Table II. Rhizone Scale Measurements W Average of Base * Base Tip and Tip Taxa L x w (mm) 'L x w (m) L x w (m) ‘Q. tomentosa 5.5 x .27 4 5 x .22 . 5.0 x .25 Range 4.5-6.0 (L) Range 5.5-6.0 (L) " .15-.54 (w) " .15-.27 (w) Q, eatonii 4.5 x .48 5.7 x .52 4.1 x .58 Range 5.2- 6. 0 (L) Range 5.0-4.5 (L) " .20—1.59(W) " .14-.55 (W) Q, eatonii f. 4.7 x .49 4.4 x .45 4.5 x .47 castanea Range 4.2—5.2 (L) Range 4.2-5.2 (L) " .15-1 0 (w) " .52-.62 (w) -)(- SpeCImen Citations in appendix. 27 Frond Stipe and Rachis The stipe of all three taxa is terete and dark reddish— brown to black. The rachis is Slightly lighter in color. Knobloch and Volz (in press) recently investigated the anatomy of the stipe and rachis of these three taxa as well as other members of Cheilanthes. They observed the following similari- ties in the genus: All stipes and rachises possess an epidermis, a cortex with an endodermis, and a stele. In the latter, one is able to discern varying amounts of pericycle, parenchyma, phloem and xylem. AS the axis matures, the basal cells of the epidermis become lignified, their lumina become small, and the cells apparently die; this series of events is of advantage to the plant in that a greater measure of support is gained. Stomates are present, being especially noticeable in the rachis. The cortex is similar in its mode of lignification. The high degree of lignification makes it difficult to obtain untorn sections. The outer cortex in some species becomes more aerenchymatous and chlorophyllous acropetal— ly. The inner cortex or endodermis is a one-celled layer of tissue with easily ruptured radial walls. The stele-cortex ratio changes acropetally with the stele occupying a proportionally smaller area up in the rachis. The vascular tissues are bounded by a pericycle which varies from one to several cells in depth. The phloem surrounds the centrally-placed xylem and consists of both Sieve and parenchyma cells. There are fewer sieve cells on the edges of the xylem than on either the adaxial or abaxial faces. Both xylem and phloem are primary tissues. Three anatomical areas of the stipe and rachis--the general cross-section, the stele, and the xylem within the stele--were examined and described by Knobloch and Volz (in press) for the taxa in this study as follows: 28 Q, eatonii- f. castanea Q, eatonii Q, tomentosa Cross-section sub-terete below, sub-terete at base sub- gradually becomes very base, terete, be- terete above changes to comes terete terete above above and in rachis Stele sulcate near the subcate below, sulcate above ground sub—sulcate and below above Xylem varies from 2- X or butter- initially 2- stranded or, at fly shape stranded or times, weakly X weak X shape condition at at base, be- base, to'X or comes strong butterfly shape X or butter- upward and into fly Shape rachis above and into rachis As Brown (1964) mentions, the Japanese worker, Ogura. (1958) "considers the 'Cheilanthes-type' of Stele to form a transitional stage between the group of ferns that he calls the ‘Monostelic' with only one stele in both the stipe and the rachis and 'Distelic' in which the stipe contains two steles that unite further up in the rachis." Thus, this may be part of the explanation for the 2-stranded xylem in some plants. Blade The length of the blade was measured from the base of the blade to the tip of the blade and the width was measured at the widest point of the blade. The stipe length was 29 measured from the base of the blade to the point of attach- ment of the stipe to the rhizome. Measurements of the blade length and width as well as the stipe length (Table III) indicate that Q. eatonii is usually shorter in length and is intermediate in width when compared to the other two taxa. Since these measurements are based on approximately 45 herbarium Specimens of each taxon, the measurements may be slightly smaller than normally found in living plants. The blade of Q, tomentosa is oblong-lanceolate to narrow- ly lanceolate and bipinnate-pinnatifid to tripinnate- tripinnatifid. The pinnae range from oblong-lanceolate to ovate-oblong with some being narrowly triangular-lanceolate. There are usually from 17 to 27 pairs of pinnae per frond. The pinnules are ovate—oblong to oblong-lanceolate with some being triangular-lanceolate. There are usually from 5 to 14 pairs of pinnules per basal pinna. The oldest pinnule arises on the side of the pinna midrib toward the apex of the frond and is therefore said to be anadromous (Mickel, 1962). The ultimate segments are suborbicular-obovate to elliptic with the terminal segment being larger than the other segments. (Plate III) The blade of Q. eatonii is oblong-lanceolate to narrowly lanceolate and tripinnate with a tripinnate-pinnatifid frond occasionally being observed. The pinnae number from 17 to 20 pairs per frond and are narrowly triangular-lanceolate to 50 ovate-oblong. The pinnules number from 5 to 9 pairs per basal pinna and are triangular-elliptic to oblong-lanceolate. The oldest pinnule is anadromous. The ultimate segments are suborbicular-oboviate to elliptic and the terminal segment is larger than the other segments (Plate IV). The blade of Q, eatonii f. castanea is oblong-lanceolate to narrowly lanceolate and tripinnate. The pinnae are narrow- ly triangular-lanceolate to triangular-ovate with some being ovate-oblong. The pinnae average from 19 to 27 pairs per frond while the pinnules average from 5 to 9 pairs per basal pinna. The pinnules are narrowly triangular-elliptic to narrowly triangular lanceolate, the oldest one being anadrom- ous. The ultimate segments are suborbicular-obovate, elliptic, or broadly ovate with the terminal segment being larger than the other segments (Plate V). Table III. Frond Measurements Blade Blade Stipe Ratio * Length Width Length Blade L: Taxa (cm) (cm) (cm) Stipe L ‘Q. tomentosa 20.2 5.8 11.6 1.7 .Q. eatonii 11.7 2.6 9.0 1.5 .Q. eatonii f. castanea 12.1 2.0 10.6 1.1 *- SpeCimen Citations in appendix. 51 Epidermal Cells To study the upper epidermal cells, imprints of the leaf surface were made using clear nail polish. All hairs were removed from the surface and a thin layer of nail polish was applied. Approximately one—half hour later or when the nail polish was dry, the imprint was peeled off by means of forceps and a dissecting needle and placed on a slide, imprint side up. A cover Slip was applied and the slide sealed. The cells were viewed and drawn by means of a microprojector. In the case of herbarium specimens, the pinnae were soaked in water with a few drops of glycerine in it to soften them. After re- maining in the softening solution overnight, the Specimens were blotted to remove excess moisture and then treated the same aS fresh material. This method was not at all successful on the lower epidermal surface. Instead, an epidermal peel was made and the cells were drawn with the aid of a camera lucida. The epidermal cells were measured from one end of the longest axis to the other and from one end of the broadest axis to the other. The stomatal measurements were made from the outer border of one guard cell to the outer border of its partner. Measurements were based on an average of 10 cells from each of five plants of each taxon. There are no tangible differences in the epidermal sur- faces of the three taxa (Plates VI and VII). The cells of the upper surface have slightly thicker walls than those of 52 the lower epidermis and are 9 to lZu longer. The thicker walls may be expected since these plants are inhabitants of dry areas. In both surfaces each cell has contact with from 5 to 8 other cells. The cells in both surfaces have moderately convoluted walls giving the general appearance of a jigsaw puzzle (Plates VI and VII). In general, there is a great deal of variation in size and shape of the cells within the same plant. Stomates are found only in the lower epidermis and are uniform in size and Shape. There may be a great deal of variation in the measurements from plant to plant as Table IV shows. The explanation according to Wagner (1952) might be as follows: "In view of the well-known capacity of laminar cells to be modified by environmental factors, the characters of the laminar cells are rather dif- ficult to evaluate precisely. That textural differences in the laminae of various ferns«are important in taxonomy is well-known, and there is little doubt that, basically, these are genetically fixed." Knobloch and Volz (1964) have investigated the leaf blade anatomy of these taxa and their findings are summarized on the following page: Q, eatonii Species f. castanea Q, eatonii Q, tomentosa Palisade Well-defined layer Cells loosely Central cells layer with few inter- arranged with rectangular, cellular spaces numerous Spaces; loosely ar- H- and Y-shaped ranged, and cells common in subquadrate central palisade in Shape on region margins of cells; H-shap- ed cells here and there Mesophyll Cells loosely Cells loosely Cells loosely layer arranged and arranged and arranged and frequently armed armed armed Vascular Large with a bun- Bundle sheath Prominent bun- bundle dle sheath; many underlain with dle sheath; tracheids; few distinct peri- tracheids cen- phloem elements and bundle paren- chyma cycle; xylem area centrally located and greater in area than phloem or parenchyma areas trally located; phloem cells few and numer- ous parenchyma cells 54 .xflpcmmmm CH coaumufio COEHOOQm * em x me me x mm #5 x mm memos mnu wo smmz mm x we me x mm mm x mm Hmaa wusnmmmq mm x aw me x aoa me x em mmlmm xoocum3 .m .m mm x He me x mm me x moa amenmm umxmm .M .b on x ow ew x 03 me x mm «mlem cmEOOOO .6 mm x we me x mm aw x mm onlmm xoocum3 .m .m mmcmummo .m.l Hflcoumm .0 mm x me «e x mm mm x Sm names may no com: mm x 05 me x mm mm x we ammmm HHOHMOO .m .D an x me 05 x mm mm x Hm mmmam coumcson .2 .H pom HHOHMOU .m .Q em x we me x mm mm x mm mwom SOOHQOGM .3 .H em x ad oe x on mm x mm maelmm unmSOOm .2 .m .I mm x Ne «w x «3 mm x mm mmn BOOHQOGM .3 .H Hflcoumm .0 pm x we mm x mu «m x mm mcwwe Onu PU com: mm x ee me x Hm mm x mm mmma SOOHQOSM .3 .H em x we mm x as mm x mm new SOOHQOGM .3 .H mm x we em x mm He x em malam mnooz .9 mm x me an x mm mm x mm meow SOOHQOG& .3 .H .l mm x #4 me x am «m x om omlmm umuumummu .m mmoucmEOu .0 Adv Adv A3 x Av Adv A3 x Av Hmnfidz HODOOHHOO mxwe mOumEOum mwfiumpflmo mHEumUHQO COHDOOHHOO * HOBOA ., HmmmD wumEOum paw mHHmU Hmanmpflmm m0 mucmEmusmmmz .>H magma 55 Hairs and Scales Hairs and scales for study were collected from the stipe, rachis, and pinna midrib of the three taxa in this study and mounted on a slide in a drop of Hoyer‘s solution. Measurements of 50 larger scales from each of the 5 areas were made from each of 4 representative specimens of each taxon. The length was measured from the tip of the scale to the base and the width of the widest part of the scale. The length:width ratio for each taxon was determined from the final measurement averages. In this study hair is defined as a structure consisting of a single cell or a single row of cells. Scale is defined as a flat plate of cells being two or more cells wide at the base and one cell thick (Foster, 1949). The rachis of Q, eatonii is covered with large scales which have a broad cordate base and gradually taper to a point at the tip (Plate VIII, Figure 8). Comparative measure- ments are given in Table V. Many smaller scales and several- celled hairs also cover the rachis (Plate IX, Figures 7 and 8). Hairs and smaller scales of the same type are found on the stipe. However, the base of the larger scales located toward the lower part of the rachis and down the stipe becomes narrower and almost truncate. No scales are found on the adaxial surface of the pinna midrib. The abaxial surface of the pinna midrib has very few scales, these being similar in shape to the scales on the apical portion of the Larger Scale Measurements Table V. Pinna Midrib wa (mm) Ratio L:W Rachis L x‘W (mm) Taxa 2.8 m~ayo 5.... “"3? “1‘“! Nt4C> Range 9.4 535 o'am “a? N30H MBfiqm .mUHm 65 g .1; .v .i. I Iur O. In rt 00.0.07. C .1 I... If. 5...“. ,1 85 a a is a ‘ E1. .1. v IdlifiJlaflla :43.» x. :53 5.x. d. .8... >H MP {Q m .‘dffla ./ 2:1 . v at: .r .313 no {mom iL « 333......- 00:6 _ .326 so... .315 3 I358 no... ......:.,u 3.5;” ulna...“ Ices-Io .5 «13-. is"; 'ci.‘ H . 66 A.C am mxumm .m amv mmODCOEOu monucmHHmno .N Ammmm smsm>o2 mummmmv mmoucmEOu mOSquHHOSU .H > MBdflm .mOHh 67 m wusmflm Dual: :3 nan... 2. any! > MBEm '- I'JCD'I II-gl a musmflm Bo $.m FIGS. 68 PLATE VI 1. Q, eatonii (Knobloch 752) upper epidermal cells 2. C. eatonii (Knobloch 752) lower epidermal cells 5. Q, eatonii f. castanea (Knobloch 65-59) upper epidermal cells 4. Q. eatonii f. castanea (Knobloch §_-5Q) lower epidermal cells 5. Q, tomentosa (Knobloch 65-69) upper epidermal cells 6. Q, tomentosa (Knobloch 6_-§Q) lower epidermal cells 69 PLATE VI Seek”... e . a. fine e we Same. 585%., $.55 ‘RWWMWQAH F M‘rflawww fireflflwfimw thmmwa mm. flew/imam“)? @555 SW 5&2me ..§ ASSESS...” we {we 1 Seam. ItswmtaatRQA: Wmmwwwwa fiefleaamw Wfiwefie . femeflMwfiMw WWAWWWSWTAW a SASS Aime. FIGS. 2. 5. 5. 7O PLATE VII Q, eatonii (Knobloch 65-49B) upper epidermal cells Q, eatonii (Knobloch 65-49B) lower epidermal cells Q, eatonii f. castanea (Knobloch 64-21) upper epidermal cells Q, eatonii f. castanea (Knobloch 64-21) lower epidermal cells Q, tomentosa (Knobloch 2048) upper epidermal cells Q, tomentosa (Knobloch 2048) lower epidermal cells 71 PLATE VII .4. Mi. .. wewaww a... e u. .3 » leeway We {5.5.5 5%.. W 5.5% MW... Newtaeeatwfiw 5% .53 1 0‘ WW H03 “55% M New w w 1 #5“ w... efiixgafiw (a fiwfiwfie :1 v) a. es. ”4:. it, a... C t m we. Seem w 2% «a .1, New... 5%... x a. . # wave, a. .1 em. 3 A a?» a J » were 9. Pinna 72 PLATE VIII .Q. tomentosa FIGS. 1. Large stipe scale 2. Large rachis scale 5. Pinna midrib scale 9, eatonii f. castanea FIGS. 4. Large stipe scale 5. Large rachis scale 6. Pinna midrib scale 9, eatonii FIGS. 7. Large stipe scale 8. Large rachis scale midrib scale 75 PLATE VIII 74 PLATE IX 9, eatonii f. castanea FIGS. 5. Pinna midrib hair Rachis hair Small rachis hair Stipe hair Small stipe scale 9, eatonii FIGS. Pinna midrib hair Rachis hair Small rachis hair Stipe hair Small stipe scale 76 PLATE X 9, tomentosa FIGS. 1. 2. 5. 4. 5. Small rachis hair Rachis hair Small stipe hair Stipe hair Pinna midrib hair BIBLIOGRAPHY Allen, R. F. 1914. Studies in spermatogenesis and apogamy in ferns. Trans. Wis. Acad. 17: 1-56. 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Some studies of apogamy in Pellaea atropurpurea (L.) Link. Trans. Am. Micro. Soc. 45: 119- 155. Holttum, R. E. 1947. A revised classification of the _Leptosporangiate ferns. Jour. Linn. Soc. Bot. 55: 125- 58. Hooker, William J. and Baker, John G. 1874. Synopsis Filicum. 2nd ed. Lond., Robert Hardwicke, p. 140. Hutchinson, A. H. 1956. The polygonal presentation of poly- phase phenomena. Proc. Trans. Roy. Soc. Canada 50: 19-26. . 1940. Polygonal graphing of ecological data. Ecology 21: 475—487. Knobloch, Irving W. 1966. Chromosome numbers in Cheilanthes and Polvpodium. Am. Jour. Bot. 55: 288-291. . In press. A preliminary review of Spore number and apogamy within the genus Cheilanthes. Am. Fern Jour. and Volz, P. A. 1964. Studies in the fern genus Cheilanthes Swartz--I. The Leaf blade anatomy of some species of the genus. Phytomorphology 14: 508-527. and . In press. Studies in the fern genus Cheilanthes Swartz--II. The anatomy of the stipes and rachises of some species. Phytomorphology. Lang, W. H. 1898. On apogamy and development of sporangia upon fern prothallia. Philos. Trans. Roy. Soc. London B. 190: 187-258. 80 Lawrence, George H. M. 1951. Taxonomy of vascular plants. New York, The Macmillan Company, 825 pp. Link, H. F. 1855. Hort. Berol. 2: 42. Manton, I. 1950. Problems of cytology and evolution in the Pteridophyta. Cambridge, University Press, 516 pp. Maxon, William R. 1919. A new Cheilanthes from Mexico. Proc. Biol. Soc. Wash. 52: 111-12. Mickel, John T. 1962. A monographic study of the fern genus Anemia, subgenus Coptophyllum. Iowa St. Jour. Sci. 56: 549-482. Morton, C. V. 1951. In_ T. H. Kearney and R. H. Peebles. Ariz. Fl. Berkeley and Los Angeles, Univ. of Calif. Press, 1052 pp. Nayar, B. K. 1965. The morphology of some species of Cheilanthes. Jour. Linn. Soc. Bot. 58: 449-460. Pichi-Sermolli, R. E. G. 1957. Adumbratio Florae Acthiopicea. 5.;Parkeriaceae, Adiantaceae, Vittariaceae. Webbia 12: 645-705. . 1960. Filicopsida. lg_Enciclopedia Agraria Italiana 4: 649-62. Steeves, Taylor A., Sussex, I. M., and Partanen, Carl R. 1955. ;g_vitro studies on abnormal growth of prothalli of the bracken fern. Am. Jour. Bot. 42: 252-244. Steil, W. N. 1919. A study of apogamy in Nephrodium hirtipes Hk. Ann. Bot., Lond. 5: 109. Stokey, Alma G. 1948. The gametophyte of Actiniopteris australis (L. fil.) Link. Jour. Indian Bot. Soc. 27: 40-49. Swanson, Carl P. 1957. Cytology and cytogenetics. Englewood Cliffs, N. J., Prentice-Hall, Inc., 596 pp. Swartz, O. 1806. Syn. Fil. 5: 196. Tyron, Rolla. 1960. A glossary of some terms relating to the fern leaf. Taxon 9: 104-109. Wagner, Warren H., Jr. 1952. The fern genus Diellia, its structure, affinities and taxonomy. Berkely and Los Angeles, Univ. of Calif. Press, 110 pp. 81 Wagner, Warren H., Jr. 1955. The genus Diellia and the value of characters in determining fern affinities. Am. Jour. Bot. 40: 54-40. . 1965. A biosystematic survey of United States ferns--preliminary abstract. Am. Fern Jour. 55: 1-16. Wherry, Edgar T. 1964. The southern fern guide. Southeastern and south-midland United States. Garden City, N. Y., Doubleday and Company, Inc., 549 pp. . 1961. The fern guide. Garden City, N. Y., Doubleday and Company, Inc., 518 pp. Whittier, D. P. 1962. The origin and development of apogamous structures in the gametophyte of Pteridium in sterile culture. Phytomorphology 12: 10-20. . 1964. The effect of sucrose on apogamy in Cyrtomium falcatum Presl. Am. Fern Jour. 54: 20-25. . 1965. Obligate apogamy in Cheilanthes tomentosa and g, alabamensis. Bot. Gaz. 126: 275-281. and Steeves, Taylor A. 1960. The induction of apogamy in the bracken fern. Canad. Jour. Bot. 58: 925-950. APPENDIX 82 APPENDIX Specimens examined - Table III, Frond Measurements* Cheilanthes eatonii UNITED STATES. Arizona: Cochise Co., limestone hill near Dos Cabezas, S, S, Phillips 2925 (US); Graham Co., Arivaipa Canyon in "The Box," S. S. Phillips 2948 (US). Colorado: El Paso Co., 17 mi. south of Colorado Springs, dry sandstone ledges east of highway south of Deadman Canyon, S, 3, Wherry S, S, (US): Las Animas Co., near Troy, S, S, Rogers 4902 (US). New Mexico: between Anton Chico and mouth of Gallinas River, S, S, Rose 17674 (US); Sandis Mts., S, Lr Herrick 987 US . Oklahoma: Cimarron Co., 4 mi. north of Kenton, S, T, Wherry s-a- (US). Texas: Brewster Co., Chisos Mts., summit of Mt. Emory, S, S, Correll 15685 (LL); mountain side of Green Gulch, C. L. Lundell 15212 (LL); Mt. Emory, S, S, Correll 15647 ITLLY; Green GuIEh} S, S, Correll 15664 (LL); near summit of Mt. Emory, S, S, Correll 15688 (LL); Basin, Chisos Mts., S, S, Correll 15615 (LL); Big Bend National Park, Cattail Falls, 2, S, Correll and S, S, Correll 50604 (LL); Basin, Chisos Mts., S, S, Correll 15614 (LL): alpine, S, S, Soxman g§1_(US): top of Pulliam Bluff Canyon, S, S, Warnock 1102 (US); Culberson Co., Sierra Diablo Mts., near summit of Victoria Peak, 2, S, Correll 15775 (LL); 10 mi. north of Van Horn, Beach Mt., S, S. Correll 15980 (LL); Sierra Diablo Mts., Victoria Canyon, S, S, Correll 15768 (LL); Guadloupe Mts., mt. stream above Pine Springs Camp, 2, S, Correll 15884 (LL); El Paso Co., Hueco Mts., Hueco Tanks, 2, S, Correll 15792 (LL); base of ledges, Hueco Tanks, 2, S, Correll 15095 (LL): Hueco Mts., Hueco Tanks, S, S, Correll 15807 (LL); Hudspeth Co., S, S, Parks S, S, (LL); Sierra Tinaja Pinta, Cornudas Range, 2, S, Correll and S, S, Correll 24705A (LL); S, S, Parks S, S, (LL); Jeff Davis Co., Davis Mts., summit of Saw- tooth Mt., S, S, Correll 15019 (LL); Morris Co., Daingerfield State Park, S, S, Correll and S, S, Correll 12480 (LL); Presidio Co., 8 mi. NW of Shafter, 2, S, Correll 15747, 15758, and 15759 (LL); Val Verde Co., near Langtry, Pump Canyon, 2, S, Correll and S, S, Correll 12915 and 12912 (LL); 100 mi. east of El Paso, Guadalupe canyon, S, S, Hitchcock 4514 (LL). *Table III is listed first because it includes most of the Specimens examined for morphological comparisons and, therefore, the specimens repeated are listed only by collector and number in the tables which follow Table III. 85 84 MEXICO. Chihuahua: 6 mi. SE of Sacramento, route #45, S, S, Correll and S, S, Johnston 217SS (LL); 5.5 mi. NW of Parral, S, S, Correll and S, S, Gentry 22716 (LL); 8 mi. NW of Parral, Minas Nuevas, 2:.§- Correll and S, S, Gentry 22749 and 22748 (LL); between Yepomera and Babicora, S, S, Correll and S, S, Johnston 21625 (LL); 15 mi. south of Encinillas, S, S. Correll 25291 (LL); 4 mi. SW of Villa Matqumoros, Sierra de Santa Barbara, S, S, Correll and S. S Gentry 22788 (LL); 12.7 mi. SW of Chihuahua on road to Cuahtemos, S. Soderstrom 916 (LL). Coahuila: 16 mi. west of Saltillo, S, S, Rollins and S, S, Tygon 58507 (US); Sierra de San Antonio, canyon at San Antonio de los Alamos, S, S, Johnston and S, S, Muller 905 (LL); Pichacho de San Jose, eastern foothills of Sierra de 1as Cruces, S, S, Stewart 1112 (LL); 1 mi. south of Carricito north facing ledge of basalt, S, S, Johnston and S, S, Muller 1&2. (LL). , Durango: Durango and vicinity, S, Palmer 895 (US). San Luis Potosi: Charcus, S, S. Lundell 5790 (US). Tamaulipas: Marcela, Stanford, Lauber, and Taylor SZL6 (US); 4 kilo. west of Miquihauana in canyon, l? S, Retherford and S, S, Northcraft 676 (US). Zacatecas: 54 mi. north of Fresnillo, S, S, Rollins andyS. S, Tyron 58257 (US); north slopes, S, S, Lloyd S, S, US . Cheilanthes eatonii f. castanea UNITED STATES. Arizona: Cochise Co., Huachuca Mts. Miller Canyon, S, S, Phillips 2801 (US). New Mexico: Carlsbad Caverns, y, Bailey §_ 2. (US). Oklahoma: Canadian Co., Devil's Canyon, S. S. Little, if: 5882 (US). Texas: Brewster Co., Chisos Mts., Green Gulch, S, S, Correll 15662, 15665.and 15667 (LL); Chisos Mts., north slope of Mt. Emory, S, S, Correll 15679, 15682, and 15654 (LL); Chisos Mts., Mt. Emory, in Basin, S, S, Correll 15646 (LL); Chisos Mts., near Basin, S. S. Correll 15642 and 15626 (LL): Chisos Mts., rimrock, C. S. Mueller 3. n. (LL); Chisos Mts., Boot Canyon, S. S. Correll 15718A (LL), _Chisos Mts., S, S, Mueller 8254 (US), Old Blue Glass Mts., S. S. Warnock 20892 (LL); Glass Mts., Gage Ranch, S. S. Warnock _20891 (LL); side Mt. Ord, Gage Estate, 11 mi. south of Alpine, S. S,N Warnock and S, McVaugh §,_S, (LL), Culberson Co.,Guadloupe Mts., SE slope of Pine Top Mt., S, S, Correll 15909 and 15887 (LL); Sierra Diablo Mts., Victoria Canyon, S. S. Correll 15758 (LL); Guadaloupe Mts., Guadaloupe Canyon, S. S. Correll 15849 (LL); Jeff Davis Co., Davis Mts., Sawtooth Mt., 2- £- Correll 15018 (LL); 10 mi. NW of Fort Davis, Sprowles Ranch, S, S. Correll 15560 (LL); Davis Mts., Limpia Canyon, S. S. Lundell and S, S, Lundell 14255 (LL); 10 mi. SE of Fort Davis, 85 tributary of Musquiz Canyon, S, S, Correll 15491A (LL); Morris Co., Daingerfield State Park, S, S, Correll and S. S. Correll 12458 (LL); Presidio Co., 8 mi. NW of Shafter, vicinity of "Elephant Rocks," S, S, Correll 15748 (LL). MEXICO. Chihuahua: other side of mts. to west from San Francisco de Oro, Arroyo de Granadefia, S, Soderstrom .SSS (LL); 15 mi. south of Encinillas, base of cliffs in canyon of large mt. mass west of route #45, S, S, Correll 25285 (LL). Coahuila: edges of Carneros Pass, S, S, Pringle S777 (LL); Saltillo, Cafion de S. Lorenzo, S. Lyonnet 5488 (LL); 11 kilo. NE of Jimulco, Ly S, Stanford, S, S. Retherford and S. S. Northcraft SS (LL); 10 mi. north of camp at La Noria, Sierra del Pino, S, S, Johnston and S, S, Muller 555 (LL): 12 mi. west of Hacienda de la Encantada, Cafion de Milagro, east side of the Sierra de los Guajes, S, S, Stewart 1711 (LL). ( ) Nuevo Leon: Hacienda Pablilla, Galeana, S, Tgylor 159 LL . Cheilanthes tomentosa UNITED STATES. Alabama: Chilton Co., ledges near Coosa River and between Mitchell Dam and Knight's Ferry, S. S. Harper 5576 (US); Cullmin Co., Mobile, S, Mohr §;_S, (US). Arizona: Santa Rita Mts., S, S, Pringle 259 (US), Bodoquivari Mts., Ly S, Goodding §-.fl- (US). Arkansas: Montgomery Co., S, Demaree 41757 (US). Georgia: Lincoln Co., 4 mi. SW of Lincolnton, Graves Mt., S, L. Wilbur and S. S.‘Webster 2801 (US); near summit of Graves Mt., S. S. Correll _and S. T. Wherry711051 (US). New Mexico: Organ Mts., S. T. S,1§.'S. (US). North Carolina: Rutherford Co., rock ridge NW of Chimney Rock Village, S, Wherry S, S, (US). Oklahoma: Johnston Co., 1 mi. east of Troy, S, S, Correll and S. S. Correll 25019 (LL); Comanche Co., Cache, S, S, Palmer 12588 (US). _South Carolina: Greenville Co., south side of Paris Mt., .S. S, Clausen and S, Trapido 5658 (US). Tennessee: Chilharvee Mts., S, Gattinger S, S, (US); Monroe Co., cliffs of Tellico River, S, S, Leeds S, S, (US). Texas: Bandera Co., head of Sabinal Canyon, S, S, Correll and S, S, Correll 12824 (LL); Brewster Co., Chisos Mts., Green Gulch, S, S, Correll 15667A (LL); Emory Peak, in basin above Laguna, C. L. Lundell 14616 (LL), Big Bend National Park, Chisos Mts., Pine Canyon, S. S. Correll and S. C. Wasshausen 27872 (LL); Chisos Mts., Mt. Emory, in basin, S. S. Correll 15648 (LL); Chisos Mts., Cat-tail Falls, S, S, -Correll and S, S, Warnock 14986 (US); Burnet Co., Inks Lake State Park, 86 .Q-.£- Lundell 15492 (LL); rock crevices on Granite Mt. near Marble Falls, S. S. Correll 15419 (LL), granite rocks near Granite Mt., S. S. Correll and S. S. Correll 12756 (LL); Cherokee Co., just beyond Jim Hogg State Park, S. S. Correll and S. S. Correll 27191 (LL); Edwards Co., on bluffs along Pulliam— Creek, near Real Co., S. S. Correll 15195 (LL); 12 mi. NW of Barksdale, "Blue Hole" on Cedar Creek, S, S_. Correll 15452 (LL), Gillespie Co., 5 mi. north of Fredericksburg, Bare #Face Rock, S. S. Correll 15255 (LL); Jeff Davis Co., 6 mi. SE of Fort —Davis, tributary of the Musquiz Canyon, S. S Correll 15502 (LL); Davis Mts., Madera Canyon, near Mt. Livermeor, S. J. Palmer 54259 (US), Davis Mts. upper slopes of Sawtooth— Mt., S, S, Correll 15011 (LL); Llano Co., NE of Baby Head, Wilbern's Glen, S, S, Correll and S, S, Correll 12700 (LL); Palo Pinto Co., 5 mi. north of Santo, S, McVaugh 8555 (LL); Tyler Co., Angelina National Forest, east of Rockland, S. S. Correll 15509 (LL); Walker Co., Huntsville, S. S. Parks S. 2- (LL) Virginia: Natural Bridge Station, S, Long and S, S, Bartram S, S, (US). West Virginia: 4 mi. SE of Charlestown, S, S, Wherry s. n. US). MEXICO. Chihuahua: Cuiteco, S, S, Knobloch 955 (MSC); 50 mi. south of Creel at Divisadero, S, S, Knobloch 847 (MSC); Moharachic, S. W. Knobloch 5972 (MSC). Nuevo Leon: Monterrey, S. Sanchez VIII-48(USL La Trinidad, Municipio De Montemorelos, S, S, Muller 2859 (LL); between Las Ajuntas and Potrero Redondo, Municipio Villa Santiago, S. S. Muller 2701 (LL), near Monterrey, ledges of Sierra Madri, _S. S. Pringle 2605 (LL) Sonora: Bavispe River, S. V. Hartman 560 (US). Tamaulipas: San Jose, above La Vegonia, S, S, Bartlett 10050 (US). 87 Specimens examined — Table I. Spore diameter Specimens previously listed under frond measurements are referred to only by collector and collection number. Cheilanthes tomentosa UNITED STATES. Alabama: Alabama Co., Blount Springs, I, y, Knobloch 2048 (MSC). * Arkansas: 2, Moore g, l,, g, 2:: accession no. 64-16 (MSG). New Mexico: Carlsbad Caverns National Park, Guadalupe Mts., E, Castetter s, 2:: accession no. 55-60 (MSC). Virginia: Ironto, I, E; Knobloch 1955 (MSC). MEXICO. ;, y; Knobloch 847 (MSC). Cheilanthes eatonii MEXICO. Q, S, Correll and I, M, Johnston 21625 (MSC): Q, S. Correll_25291 (MSC). Chihuahua: Picacho de San Jose, g, M, Stewart 1112, I.‘W. Knoblochls accession no. 65-49B (MSC); in canyon SE of Hidalgo de Parral. I, E} Knobloch 752 (MSC). Cheilanthes eatonii f. castanea UNITED STATES. New Mexico: Carlsbad Caverns National Park, g, 5, Baker 5, 2,, accession no. 64-21 (MSC). Oklahoma: Caddo Co., Caddo Canyons near Hinton, IQ. Goodman g, Q,, accession no. 65-49D (MSC). Texas: Davis Mts., g, g, Warnock s, g,, accession nos. 63-28 and 65-30 (MSC). MEXICO. Lefebure 1161, accession no. 63-52 (MSC). 88 Specimens examined - Table II. Rhizome Scale Measurements Cheilanthes tomentosa UNITED STATES. Texas: Llano Co., Turkey Peak, 2, g, Correll and H. B. Correll 12771 (LL); Gillespie Co., Bear Mt., H. B. Parks 5. n. (LL); 2, g, Correll and M, M, Warnock 14986- (LL); C. L. _Lundell 14616 (LL); 2, g, Correll and H. B. Correll 12700 (LL); Q..M, Muller 2101 (LL). Cheilanthes eatonii MEXICO. Coahuila: San Jose, base of Sierra de las Cruces, I. M. Johnston and C. H. Muller 980 (LL) Q. L. Lundell 13212 (LL): D. s. Correll 15647.15768, and 15759 (LL); C. H. Muller 162 (LL); H. M_. Parks s, 2: (LL); Q. L. Hitchcock :514 (LL) Cheilanthes eatonii f. castanea UNITED STATES. Texas: Brewster Co., north slope of Mt. Emory, Chisos Mts., M, M, Correll 15678 (LL). MEXICO. Sierra de la Madera, Cafion del Agua, Cuatro Ciengas Municipiode, Q, M, Muller 5252B (LL). D. S. Correll 15642, 15682, 15748, and 1§663 (LL); Q. L. _Lundell and A.‘ A. Lundell 14255 (LL): Q, Q, Pringle 2777 (LL). 89 Specimens examined - Table IV. Measurements of Epidermal Cells and Stomata Cheilanthes tomentosa MEXICO. Nuevo Leon: Cafion de San Francisco, ;, M, Knobloch 1966 (MSC). ;, M, Knobloch 2048, and Mil (MSC): M, Moore s. M., accession no. 61-16 (MSC); M. Castetter s, 2,, accession no. 63-60 (MSC). Cheilanthes eatonii MEXICO. Chihuahua: El Kilo Mt., south of Juarez, L, M, Knobloch 2062 (MSC) D. S. Qorrell and I. JM. Johnston 21625 (MSC); R. M. Stewart 1112 I. W. Knobloch' s accession no. 65- 49B (MSC); .1. QM. Knobloch 752 (MSC);'Q. .s. Correll 23291 (MSC)- Cheilanthes eatonii f. castanea Lefebure 1161 (MSC): M. M, Warnock s, 2:: accession nos. 65-28 and 65-50 (MSC): Q, Goodman g, 2:: accession no. 64-21 (MSC): g, M, Baker g, M,, accession no. 63-49B (MSC). Specimens examined - Table V. Larger Scale Measurements Cheilanthes eatonii D. S. Correll 25291 and 15759 (LL): T. Soderstrom 916 (LL); R. M. Stewart 1112 (MSC). Cheilanthes eatonii f. castanea B. H. Warnock 20891 (LL); D. s. Correll 13718A, 15682, and 25285 (LL) Cheilanthes tomentosa c. L. Lundell 14616 (LL); 2, 27872_ (LL): R. McVaugh 8555 (LL): Correll and Wasshausen 'M. Muller 2701 (LL). Rum MICHIGAN STATE UNIVERSITY LIB A I III H III III IIIIIIES O 3015 3497 3 1293