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A Study of the Germination Process of Seeds of Heteranthera Limosa. James Earl Marler Louisiana State University and Agricultural & Mechanical College

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A Study of the Germination Process of Seeds of Heteranthera Limosa. James Earl Marler Louisiana State University and Agricultural & Mechanical College Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1969 A Study of the Germination Process of Seeds of Heteranthera Limosa. James Earl Marler Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Marler, James Earl, "A Study of the Germination Process of Seeds of Heteranthera Limosa." (1969). LSU Historical Dissertations and Theses. 1607. https://digitalcommons.lsu.edu/gradschool_disstheses/1607 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. This dissertation has been microfilmed exactly as received 70-254 MARLER, James Earl, 1939- A STUDY OF THE GERMINATION PROCESS OF SEEDS OF HETERANTHERA LIMQSA. The Louisiana State University and Agricultural and Mechanical College, Ph*D., 1969 Botany University Microfilms, Inc., Ann Arbor, Michigan A Study of the Germination Process of Seeds of Heteranthera limosa. A Dissertation Submitted to the Graduate School of the Louisiana State University Agriculture and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Botany and Plant Pathology by James Earl Marler B.S., University of Miami, 1962 M.A., University of Texas, 1965 May, 1969 ACKNOWLEDGEMENT The author wishes to express his gratitude to Dr. John B. Baker for his guidance, patience, and encouragement dur­ ing these investigations and also during the preparation of this dissertation. He also acknowledges the aid and encouragement of the members of his committee. Special thanks go to Angela S. Blaize and Linda J. Rebowe for their excellent technical assistance. The author extends special acknowledgement to his wife, Harriet, for her technical assistance in the preparation of this manuscript. ii TABLE OF CONTENTS Page ACKNOWLEDGEMENT............................... ii LIST OF TABLES ............................... v LIST OF FIGURES.......... vii ABSTRACT..................................... ix INTRODUCTION................................. 1 LITERATURE REVIEW............................. 3 METHODS AND MATERIALS......................... 17 General ................. ........ 17 Experimental organism.............. 17 Plant propagation and seed production....................... 17 Chemicals and Gases ..................... 18 Standard Germination Conditions and Procedures......................... 19 Respiration and Inhibitor Experiments . 19 Embryo Isolation Experiments............ 21 Oxygen and Nitrogen Mixture ............ 22 Light Response Study.......... 22 Light quality experiments.......... 22 Red/far-red experiments............ 22 Histological Study....................... 24 Fixing, embedding, sectioning. 24 Stains and staining procedure. 25 Phase-Contrast Microscope Study ........ 26 RESULTS....................................... 27 Studies on the Effect of Oxygen on Germination......................... 27 Studies on the Light Requirement for Germination . ....................... 28 Studies on the Effect of Cracking the Seed C o a t ................ 33 Studies on the Effect of Temperature on Germination......................... 36 iii Page Studies on the Induction of Germination by Various Chemicals . 38 Respiration Studies...................... 40 The Effect of Reducing Agents on Germination.......................... 45 Histological Studies .................... 47 DISCUSSION..................................... 67 SUMMARY ....................................... 78 LITERATURE CITED.............................. 81 VITA 92 LIST OF TABLES TABLE Page I. The influence of various gases on the germination of seed of H. limosa............... 29 2. The effect of photoperiod and relative light intensity on germination and the time required for maximum germination .... 29 3. The comparison of continuous and in­ terrupted red light in the stimulation of germination of seed of H. limosa under standard germination conditions .... 32 4. The effect of light wavelength ohjthe germination of seed of H. limosa. .......... 32 5. The effect of red (660 mp) and far- red (750 mp) light on the germination of seed of H. limosa......................... 34 6 . The minimal requirements of light and anaerobic conditions for germination of seed of H. limosa......................... 34 7. The effect of separation of the anaerobic and light requirement on the germination of seed of H. limosa......................... 35 8 . The effect of cracking the seed coat on the germination of seed of"H. limosa in air, light and d a r k ......................... 35 9. The growth of excised embryos on nutrient agar in air, Ng, light, and dark............ 37 10. The effect of germination stimulators on the germination of H. limosa under un­ favorable conditions......................... 37 11. The effect of metabolic inhibitors on the germination of whole and cracked seed .... 43 v TABLE Page 12. The effect of reducing agents on the germination of H. limosa in three con­ centrations of oxygen under standard light and temperature conditions............ 46 13. The germination of Typha latifolia (cattail) under various germination conditions ............................... 65 vi LIST OF FIGURES FIGURE Pa fee 1. The vessel used for the germination of seed of H. limosa........................... 20 2. Transmission spectra of various cellophane filters ......................... 23 3. The effect of various concentrations of oxygen (in N?) on the germination of H. limosa s e e d s ......................... 30 4. The effect of various constant tem­ peratures on the germination of seed of H. limosa................................ 39 5. The inhibition of anaerobic respira­ tion of seed of H. limosa with 10“3m iodoacetate................................. 41 6 . The uptake of oxygen by seeds in air, under standard light and temperature conditions................................. 42 7. The effect of metabolic inhibitors on the anaerobic respiration of seed of H. limosa under standard germination conditions ................. 44 8 . Whole germinating seed of H. limosa showing the emerging radicle and the trap-door through which it emerged ........ 48 9. Longitudinal section of the seed of H. limosa at 100 X ......................... 49 10. Plastic-embedded section of seed showing the structure of the seed coat ........................ 52 11. Dorsal view of a portion of the t r a p - d o o r ........................... 53 vii FIGURE Page 12. The trap-door of H-s. limosa at zero time showing the closely packed cells of the seed coat........................... 54 13. The trap-door of H. limosa after 24 hours incubation under germination conditions................................. 56 14. The trap-door of H. limosa after 48 hours incubation under germination conditions................................. 57 15. The trap-door of H. limosa at about 72 hours of' incubation..................... 58 16. The trap-door of H. limosa at 72-84 hours of incubation....................... 59 17. The aleurone grains of H. limosa........ 61 18. The chalazal end of the embryo of £• limosa showing the disappearance of the aleurone g r a i n s . ................... 62 19. Embryo of H. limosa about 12 hours after germination ....................... 63 20. Germinating seed of Typha latifolia In air showing the emergence of the radicle and the trap-door (arrow)........ 64 viii ABSTRACT The seed of the aquatic plant, Heteranthera limosa Willd., are seen to have two requirements for germination: anaerobic conditions and light. Dormancy involves the seed coat which acts as a mechanical barrier to germination. The light requirement appears to be a phytochrome mediated system which is unique in that it is unaffected by anaerobic conditions. The seed obtains its energy through fermentative re­ spiration and seedling growth is unaffected by 10“^M cyanide. Iodoacetate completely blocks germination and seedling growth. Reducing agents are able to substitute for anaerobic condition if the O2 concentration is below 10%. Reduced glutathione, cysteine, thiosulfate, mercaptoethanol, and mercaptoacetate were all capable of stimulating germination. Histological studies show a progressive breakdown of intercellular material along a cleavage line of a trap­ door through which the radicle emerges. The force re­ quired to open the weakened trap-door is provided by the expansion of certain aleurone grain containing cells of the embryo cotyledon. INTRODUCTION This dissertation originally began as a weed control study of an occasionally bothersome aquatic weed in rice, Heteranthera limosa. The seeds of this plant were field harvested and brought into the laboratory for seedling studies. However, it was immediately evident that the seeds would not germinate on moist filter paper at any temperature. When the seeds were placed on top of soil or sand and flooded with water, they geminated readily provided the water was not aerated. Further preliminary study indicated the seeds required an oxygen limited system and white light to geminate. A study was begun to determine the nature of dormancy involved and a possible physiological explanation of the process of gemination. Domancy can be imposed upon a seed by internal or ex­ ternal factors. Internal factors responsible
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