Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1983 A light and electron microscopic study of the development of antheridia in Onoclea sensibilis L Jane Louise Kotenko Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Botany Commons Recommended Citation Kotenko, Jane Louise, "A light and electron microscopic study of the development of antheridia in Onoclea sensibilis L " (1983). Retrospective Theses and Dissertations. 8494. https://lib.dr.iastate.edu/rtd/8494 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. 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Uni International 300 N. Zeeb Road Ann Arbor, Ml 48106 8407093 Kotenko, Jane Louise A LIGHT AND ELECTRON MICROSCOPIC STUDY OF THE DEVELOPMENT OF ANTHERIDIA IN ONOCLEA SENSIBILIS L (VOLUMES I AND II) Iowa State University PH.D. 1983 University Microfilms IntGrnâtiOnâl 300 N. Zeeb Road, Ann Arbor, Ml 48106 PLEASE NOTE: In all cases this material heis been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark V . 1. Glossy photographs or pages 2. Colored illustrations, paper or print 3. Photographs with dark background 4. Illustrations are poor copy 5. Pages with black marks, not original copy 6. Print shows through as there is text on both sides of page 7. Indistinct, broken or small print on several pages \X 8. Print exceeds margin requirements 9. Tightly bound copy with print lost in spine 10. Computer printout pages with indistinct print 11. Page(s) lacking when material received, and not available from school or author. 12. Page(s) seem to be missing in numbering only as text follows. 13. Two pages numbered . Text follows. 14. Curling and wrinkled pages 15. Other University Microfilms International A light and electron microscopic study of the development of antheridia in Onoclea sensibilis L. by Jane Louise Kotenko Volume 1 of 2 Â Dissertation Submitted to the Graduate Faculty in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY Major: Botany Approved : Signature was redacted for privacy. In Charge of Major Work Signature was redacted for privacy. For the Major Depaj^roient Signature was redacted for privacy. For the Gradi fee College Iowa State University Ames, Iowa 1983 ii TABLE OF CONTENTS Page PREFACE iv INTRODUCTION 1 LITERATURE REVIEW 5 Taxonomic Classification 5 Gametophyte Morphology 5 Cellular Anatomy of Fern Gametophytes 10 Antheridiogens and Antheridium Formation 15 Location of Antheridium Initials 18 Antheridium Morphology 21 Spermatogenesis 32 MATERIALS AND METHODS 35 Culture Conditions 35 Light Microscopy (LM) 40 Transmission Electron Microscopy (TEM) 47 Scanning Electron Microscopy (SEM) 47 RESULTS 49 Preliminary Experiments 49 Gametophyte Growth Data 55 Antheridium Developmental Data 72 Vegetative Cell Development and Structure 113 Cellular Changes during the Division Phase of Antheridium Initiation and Development 121 Cellular Changes during the Differentiation of Spermatids into Spermatozoids 161 DISCUSSION AND CONCLUSIONS 171 ill Growth of Gametophytes 171 Induction and Location of Initials In Relation to Gameto- phyte Physiology 174 Formation of Initials from Asymmetric Cell Divisions 182 Comparative Antheridium Morphology 191 Spermatogenesis 209 BIBLIOGRAPHY 226 ACKNOWLEDGMENTS 241 APPENDIX A: KEY TO LABELING 242 APPENDIX B: FIGURES 244 APPENDIX C: CULTURE MEDIUM PREPARATION 419 APPENDIX D: GAMETOPHYTE CLEARING TECHNIQUE AND METHODS OF DATA COLLECTION 421 Clearing Technique and Slide Preparation 421 Gametophyte Collection 421 Data Collection and Analysis 421 APPENDIX E: FORMULAE AND SCHEDULES 425 APPENDIX F: MEAN VALUES AND CONFIDENCE INTERVALS 431 iv PREFACE This dissertation is dedicated to the memory of Gregory Bateson, anthropologist/psychologist/cyberneticist, son of William Bateson, geneticist. Bateson spent his whole life searching for the "pattern which connects" all living creatures with each other. My interests, either directly or indirectly, have involved studying the continuum or cycling of life. A fundamental question concerns how an organism can recreate itself by producing two very specialized cells, the sperm and the egg. This present study was conducted to investigate part of a fern life cycle; in particular, the formation of male gametangia and the eventual formation of sperms. I have experienced similar feelings to those ex­ pressed by Lewis Thomas (1974), Chancellor of Sloan-Kettering Cancer Institute, as he wrote about human embryology. "For the real amaze­ ment, if you want to be amazed, is the process. You start out as a single cell derived from the coupling of a sperm and an egg, this divides into two, then four, then eight, and so on, and at a certain stage there emerges a single cell which will have as all its progeny, the human brain. The mere existence of that cell should be one of the great astonishments of the earth. People ought to be walking around all day, all through their waking hours, calling to each other in endless wonderment, talking of nothing except that cell." To be realistic, my amazement of living systems is often tempered with everyday frustrations and pressures, spawning self-doubts about the importance of science. Lewis Thomas (1974) has been able to place V into perspective for me the phenomena of science and scientists. "The most mysterious aspect of... science is the way it is done. Not the routine, not just the fitting together of things that no one had guessed at fitting, not the making of connections; these are merely the workaday details, the methods of operating. They are interesting, but not as fascinating as the central mystery, which Is that we do it at all, and that we do it under such compulsion.... It sometimes looks like a lonely activity, but it is as much the opposite of lonely as human behavior can be. There is nothing so social, so communal, so inter­ dependent.... It is instinctive behavior in my view, and I do not under­ stand how it works.... While it is going on, it looks and feels like aggression: get at it, uncover it, bring it out, grab it, it's mine I It is like a primitive running hunt, but there is nothing at the end to be injured. More probably the end is a sigh. But then, if the air is right and the science is going well, the sigh is immediately inter­ rupted, there is a yawping new question, and the wild, tumbling activity begins once more, out of control all over again." I believe that being a scientist is Instinctual, for I have ex­ perienced total dissolution of frustrations during the discovery of a new developmental stage. When I wonder whether my work is significant or not, I think of the answer given by Gandhi. "Whatever you do will be insignificant, but it is very Important that you do it." 1 INTRODUCTION This study was designed to investigate, using light and electron micros­ copy, the morphological and anatomical changes which occur during the ini­ tiation and subsequent development of sperm-producing structures known as antheridia. All sexually reproducing homosporous plants form antheridia during the gametophytic stage of their life cycle, but antheridium morphol­ ogy and mode of development vary greatly among different taxonomic groups. Onoclea sensibilis L.. an advanced, leptosporangiate fern was selected as the experimental organism in this study due to the simplicity of its antheridia, composed of only three sterile, jacket cells sur­ rounding the spermatogenous tissue. Each antheridium arises from an asymmetric cell division in one of the vegetative cells of a morpho­ logically simple gametophyte plant. The fast-growing gametophytes are less than 1 mm in length and width at sexual maturity and are composed of only one layer of chlorenchymatous vegetative cells.