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Use of Micropropagation Techniques to Improve Germination

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Use of Micropropagation Techniques to Improve Germination USE OF MICROPROPAGATION TECHNIQUES TO IMPROVE GERMINATION SUCCESS IN SIX SPECIES OF CACTI _____________ A Thesis Presented to The Faculty of the Department of Biological Sciences Sam Houston State University _____________ In Partial Fulfillment of the Requirements for the Degree of Master of Science _____________ by David Warren-Hammack May, 2019 USE OF MICROPROPAGATION TECHNIQUES TO IMPROVE GERMINATION SUCCESS IN SIX SPECIES OF CACTI by David Warren-Hammack ______________ APPROVED: Chris Randle, PhD Thesis Director John Pascarella, PhD Committee Member Chad Hargrave, PhD Committee Member John Pascarella, PhD Dean, College of Science and Engineering Technology DEDICATION I dedicate this thesis to my family. My dad, Stewart Simon, for being supportive and an influence in my efforts towards my life goals and my person. My sister, Lesha, for being my best friend while I was in my worst times. My mother, Cindy Simon, who gave part of herself for me, literally, so I could survive and for fighting for me so I could fight for myself. And my husband, Jeremy Warren, who, when I needed a dose of reality or a shoulder, I can always rely on for anything. iii ABSTRACT Warren-Hammack, David , Use of micropropagation techniques to improve germination success in six species of cacti. Master of Science (Biology), May, 2019, Sam Houston State University, Huntsville, Texas. Cacti have been a major contributor to the ecosystem of desert fauna as a constant food source during dry seasons. However, this resource has reached a point of dwindling due to many human interferences. In situ conservative efforts have not been stable enough to maintain the efforts of preserving population genetics. Therefore, in vitro techniques will be required to counter the effects. Following previous studies, micropropagation techniques were analyzed to optimize germination number, time, and rate around three variables; difference in nutrient media, gibberellic acids, and species. Two trials were run at intervals of eight weeks; the second trial a few weeks after the first had ended. Results showed a strong significance in emergence and germination affected by species type for both trials. There were other significant factors including interactions between variables. Overall, this experiment showed overwhelming evidence towards the need to treat species to separate protocols in micropropagation techniques. KEY WORDS: Cactus, Micropropagation, Botany, Conservation, Seed, Germination techniques, Sam Houston State University, Texas iv ACKNOWLEDGEMENTS I would like to acknowledge my team at the Randle lab at Sam Houston State University; specifically, Austin Brenek, Kayla Hankins, Christian McMurray, Sana Sadiq, Andrew Martinez, Jesse Rodriguez, and Reagan King for their assistance in plating and counting seeds and materials for this experiment. I would also acknowledge Dr. Joan Hudson for her knowledge and helping me acquire materials that I could not afford the time or the money to get. A huge thank you to Gloria Randle for her help in plating, counting seeds, and preparing materials for the experiment. Without you I could not have done this. v TABLE OF CONTENTS Page DEDICATION ................................................................................................................... iii ABSTRACT ....................................................................................................................... iv ACKNOWLEDGEMENTS ................................................................................................ v TABLE OF CONTENTS ................................................................................................... vi LIST OF TABLES ........................................................................................................... viii LIST OF FIGURES ........................................................................................................... ix CHAPTER I INTRODUCTION .................................................................................................. 1 Ecological Functions ............................................................................................... 2 Conservation ........................................................................................................... 4 Solutions ................................................................................................................. 7 Study Questions .................................................................................................... 11 REFERENCES ..................................................................................................... 12 II METHODS ........................................................................................................... 22 Substrate Preparation ............................................................................................ 25 Sterilization ........................................................................................................... 25 Measurements and Analysis ................................................................................. 26 REFERENCES ..................................................................................................... 29 III RESULTS ............................................................................................................. 32 Trial 1 .................................................................................................................... 34 Trial 2 .................................................................................................................... 45 vi IV DISCUSSION ....................................................................................................... 49 Emergence............................................................................................................. 50 Germination .......................................................................................................... 52 Problems ............................................................................................................... 53 Conclusions ........................................................................................................... 54 REFERENCES ..................................................................................................... 55 REFERENCES ................................................................................................................. 56 VITA ................................................................................................................................. 68 vii LIST OF TABLES Table Page 1 Experimental Variables in Type ........................................................................... 27 2 ANOVA results of emergence with Trial block ................................................... 33 3 ANOVA results of germination with Trial block ................................................. 33 4 ANOVA results of Trial 1 emergence .................................................................. 34 5 Emergence contrast between Species with respect to Media ................................ 36 6 Emergence contrast between Hormones with respect to Media ........................... 37 7 ANOVA results of Trial 1 germination ................................................................ 40 8 Germination contrast between Species with respect to Hormone ........................ 41 9 Germination contrast between Species with respect to Media ............................. 42 10 Germination contrast between Hormone with respect to Media .......................... 43 11 ANOVA results of Trial 2 emergence .................................................................. 46 12 ANOVA results of Trial 2 germination ................................................................ 47 viii LIST OF FIGURES Figure Page 1 Oreocereus celsianus (Cactaceae) cut latitudinally. ............................................... 3 2 Distribution of Trial 1 emergence by species. ...................................................... 35 3 Distribution of Trial 1 emergence by hormone concentration. ............................. 35 4 Species versus media least significant difference of Trial 1 emergence. ............. 37 5 Hormone versus media least significant difference of Trial 1 emergence. ........... 38 6 Species, media, and hormone least significant difference of Trial 1 emergence. ............................................................................................................ 39 7 Distribution of Trial 1 germination by species. .................................................... 40 8 Distribution of Trial 1 germination by media. ...................................................... 41 9 Species versus hormone least significant difference of Trial 1 germination. ....... 42 10 Species versus media least significant difference of Trial 1 germination. ........... 43 11 Hormone versus media least significant difference of Trial 1 germination. ......... 44 12 Species, media, and hormone least significant difference of Trial 1 germination. .......................................................................................................... 44 13 Emergence and Germination Differences Trials 1 and 2. ..................................... 45 14 Distribution of Trial 2 emergence by species. ...................................................... 46 15 Distribution of Trial 2 emergence by media. ........................................................ 47 16 Distribution of Trial 2 germination
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