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In Vitro Regeneration of Cladrastis Kentukea (American Yellowwood) and Cornus Kousa (Kousa Dogwood)

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In Vitro Regeneration of Cladrastis Kentukea (American Yellowwood) and Cornus Kousa (Kousa Dogwood) University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Masters Theses Graduate School 5-2005 In Vitro Regeneration of Cladrastis kentukea (American yellowwood) and Cornus kousa (kousa dogwood) Denita Hadziabdic University of Tennessee - Knoxville Follow this and additional works at: https://trace.tennessee.edu/utk_gradthes Part of the Entomology Commons Recommended Citation Hadziabdic, Denita, "In Vitro Regeneration of Cladrastis kentukea (American yellowwood) and Cornus kousa (kousa dogwood). " Master's Thesis, University of Tennessee, 2005. https://trace.tennessee.edu/utk_gradthes/1864 This Thesis is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Masters Theses by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a thesis written by Denita Hadziabdic entitled "In Vitro Regeneration of Cladrastis kentukea (American yellowwood) and Cornus kousa (kousa dogwood)." I have examined the final electronic copy of this thesis for form and content and recommend that it be accepted in partial fulfillment of the equirr ements for the degree of Master of Science, with a major in Entomology and Plant Pathology. Robert N. Trigiano, Major Professor We have read this thesis and recommend its acceptance: Dr. Stephen Garton, Dr. Mark T. Windham Accepted for the Council: Carolyn R. Hodges Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) To the Graduate Council: I am submitting herewith a thesis written by Denita Hadziabdic entitled “In Vitro Regeneration of Cladrastis kentukea (American yellowwood) and Cornus kousa (kousa dogwood). “ I have examined the final electronic copy of this thesis for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Master of Science, with a major in Entomology and Plant Pathology. Robert N. Trigiano_______ Major Professor We have read this thesis and recommend its acceptance: Dr. Stephen Garton Dr. Mark T. Windham Accepted for the Council: Anne Mayhew______________ Vice Chancellor and Dean of Graduate Studies (Original signatures are on file with official student records.) In Vitro Regeneration of Cladrastis kentukea (American yellowwood) and Cornus kousa (kousa dogwood) A Thesis Presented for the Master of Science Degree The University of Tennessee Denita Hadziabdic May 2005 DEDICATION I would like to dedicate this work to my dad, the late Abdurahman Hadziabdic for his unconditional love, support, and never-ending faith in me. I wish you were here to share the joy of this incredible journey with me so I could, once again, make you proud. Thank you for beautiful memories. ii ACKOWLEDGMENTS I would like to express my sincere gratitude to my major professor, Dr. Robert N. Trigiano for his guidance, encouragement, and above all, his friendship during my graduate study here at the University of Tennessee. This would have been a totally different experience and definitely less enjoyable without you. I would like to thank my other committee members Dr. Stephen Garton and Dr. Mark T. Windham for their help on this project and valuable suggestions. It was my true pleasure to get to know you. Special thanks to Dr. William E. Klingeman for his help with statistical analysis. My deepest thanks to Melissa H. Ament for her friendship, guidance, and great sense of humor, which have made the lab a great working environment and wonderful place to be. I am also very thankful to David McCammon for his help in the greenhouse and willingness to assist anytime, regardless of how busy he was. Thanks to everyone in the department of Entomology and Plant Pathology, my fellow graduate students, and everyone who made my life wonderful, as well as miserable at the time. It has been a true pleasure working with all of you. Most importantly, I would like to thank my family, my mom, Sefika, for her selfless love and support throughout all these years, my sisters, Sanela Sahinovic and Enida Islamagaic, as well as their families. Without your iii encouragement, love and friendship I would not be writing these words today. I feel extremely blessed to have you in my life. Thank you for allowing me to follow my dreams. Thanks to Karen and Steve Case and Sue Ellen Carter for being my adopted American family whom I will always be deeply and sincerely grateful. Without you, my life would be completely different. I would like to sincerely thank Dr. Gary Bachman for his guidance during my undergraduate studies. I enjoyed working with you and loved your enthusiasm to share knowledge with students. You taught me to love research in a different and strange way, and I am here because of your faith in me. I would also like to use this opportunity and thank everyone who helped me in any way during my undergraduate and graduate studies. I am extremely grateful for all your assistance and hopefully, one day I will be able to help someone eager to chase their dreams, just like I did. Lastly, thanks to Matthew T. Johnson for his love, friendship and above all, his unwavering moral support during times I needed him the most. I am very appreciative of many hours you spent proofreading this work. Thank you for being there for me through all this time. Without you, this experience would not be as enjoyable and pleasurable as it is. iv ABSTRACT Selection of superior individuals followed by clonal vegetative propagation is a very important strategy for plant improvement. Cloning via tissue culture can produce a population of an identical genotype without limits. A single specimen tree of Cladrastis kentukea and five different Cornus kousa cultivars were selected for tissue culture studies. These trees exhibited superior performance in horticultural trials, including disease resistance and drought tolerance, which are highly important to the green industry. Axillary buds from a single C. kentukea tree were initially cultured on either Woody Plant Medium (WPM) or Murashige and Skoog (MS) containing 0, 1, 2, or 4 µM 6–benzylaminopurine (BA). Cultures were transferred to fresh medium every four weeks. Elongated shoots were harvested after thirty-nine weeks and transferred to half-strength MS medium supplemented with following concentrations of IBA: 0, 3, 30, 100, and 300 µM for three days then returned to half-strength MS without growth regulators. Explants exposed to 300 µM of IBA produced significantly more roots (75%) compared to explants exposed to other treatments. Fifty- four and forty- six percent of the microshoots rooted when exposed to 100 and 30 µM IBA, respectively. Only 4% of the microshoots rooted when exposed to 3 µM IBA and none of the microshoots in the control treatment (0 IBA) rooted. Although 300 µM treatment yielded the most rooted plantlets, there was significantly higher terminal meristem abortion compared to other treatments. There were no statistical differences between the numbers of roots v and total root length among all treatments. Additionally, all microshoots that rooted had lenticels, suggesting that presence of lenticel cambial activity can possibly predict rooting abilities of selected microshoots. Rooted microshoots were gradually acclimatized to non-sterile environment. Axillary and apical buds from five Cornus kousa cultivars (‘Little Beauty’, ‘Samaritan’, ‘Heart Throb’, ‘Rosabella’ and ‘Christian Prince’) were initially established on either WPM or one-half Woody Plant Medium/Broad Leaved Tree Medium (BW), amended with the following concentrations of 6– benzylaminopurine (BA): 0, 2, 4, and 8 µM. After explants were transferred at four-week intervals for 28 weeks beginning in April, only microshoots of ‘Samaritan’, ‘Heart Throb’, and ‘Rosabella’, were harvested from proliferating cultures and placed on rooting media. ‘Little Beauty’ and ‘Christian Prince’ did not perform well in multiplication phase of tissue culture and were excluded from further studies. Rooting media contained WPM or BW supplemented with either 1- naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), or indole-3-acetic acid (IAA) at the following concentrations: 0, 0.5, 1.5, 4.5, and 13.5 µM. Six weeks following rooting experiment, preliminary data was collected and results indicated a total of nine plants rooted on both WPM and BW media supplemented with IBA, 17 plants rooted on media supplemented with NAA, and 14 plants rooted plants supplemented with IAA. NAA and IAA appeared to be better for root production on C. kousa cultivars microshoots than IBA. Moreover, both WPM and BW media supported rooting of C. kousa microshoots. However, WPM appears to support more root production compared to BW. A greater vi number of ’Samaritan’ and ‘Heart Throb’ microshoots rooted on WPM amended with a wide range of NAA concentrations, whereas more ‘Rosabella’ microshoots rooted on BW medium amended with various concentrations of IAA. Since ‘Rosabella’ and ‘Heart Throb’ are very closely related and should have rooted with similar treatments, further research is needed to confirm this finding. Additionally, microshoots placed on either basal media supplemented with NAA produced significant amount of callus compared to microshoots exposed to other growth regulator treatments. The highest mean number of roots per rooted microshoot was recorded on ‘Samaritan’ when exposed to various NAA concentrations.
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