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The Role of Gibberellic Acid in Aphid-Plant-Arbuscular Mycorrhizal Fungus Interactions

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The Role of Gibberellic Acid in Aphid-Plant-Arbuscular Mycorrhizal Fungus Interactions University of Northern Colorado Scholarship & Creative Works @ Digital UNC Master's Theses Student Research 5-8-2020 THE ROLE OF GIBBERELLIC ACID IN APHID-PLANT-ARBUSCULAR MYCORRHIZAL FUNGUS INTERACTIONS Drew Olson [email protected] Follow this and additional works at: https://digscholarship.unco.edu/theses Recommended Citation Olson, Drew, "THE ROLE OF GIBBERELLIC ACID IN APHID-PLANT-ARBUSCULAR MYCORRHIZAL FUNGUS INTERACTIONS" (2020). Master's Theses. 164. https://digscholarship.unco.edu/theses/164 This Dissertation/Thesis is brought to you for free and open access by the Student Research at Scholarship & Creative Works @ Digital UNC. It has been accepted for inclusion in Master's Theses by an authorized administrator of Scholarship & Creative Works @ Digital UNC. For more information, please contact [email protected]. © 2020 Drew Olson ALL RIGHTS RESERVED UNIVERSITY OF NORTHERN COLORADO Greeley, Colorado The Graduate School THE ROLE OF GIBBERELLIC ACID IN APHID- PLANT-ARBUSCULAR MYCORRHIZAL FUNGUS INTERACTIONS A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science Drew Olson College of Natural and Health Sciences School of Biological Sciences May 2020 This Thesis by Drew Olson Entitled: The role of gibberellic acid in aphid-plant-arbuscular mycorrhizal fungus interactions has been approved as meeting the requirement for the Degree of Master of Science in College of Natural and Health Sciences in School of Biological Sciences, Program of Biological Sciences Accepted by the Thesis Committee: Susana Karen Gomez, Ph.D., Chair Mitchell McGlaughlin, Ph.D., Committee Member Hua Zhao, Ph.D., Committee Member Accepted by the Graduate School Cindy Wesley, Ph.D. Interim Associate Provost and Dean Graduate School and International Admissions ABSTRACT Olson, Drew. The role of gibberellic acid in aphid-plant-arbuscular mycorrhizal fungus interactions. Unpublished Master of Science thesis, University of Northern Colorado, 2020. Arbuscular mycorrhizal (AM) fungi form beneficial associations with the roots of most terrestrial plants, which is mostly characterized by increased plant nutrient acquisition (phosphate and nitrogen) by the fungi in exchange for carbon and lipids from the plants. These associations can also enhance plant coping mechanisms to abiotic and biotic stresses. However, insect herbivores, such as aphids, can benefit from AM fungi- plant symbioses by altering plant defenses that are driven mainly by the phytohormones jasmonic acid (JA) and salicylic acid (SA). Nevertheless, other phytohormones can modulate plant defenses by altering the balance of the JA and SA signaling pathways, such as the growth promoting phytohormone gibberellic acid (GA). Although GA signaling plays an important role in modulating plant defenses during plant-pathogen interactions, its role in insect-plant and insect-plant-beneficial microbe interactions remains largely unknown. Therefore, the current study set out to investigate the following two objectives: 1) examine the role of exogenous GA application in regulating plant defenses during aphid-plant-AM fungus interactions, and 2) evaluate the role of GA signaling in modulating plant defenses during aphid-plant interactions. In objective 1, we found that exogenous GA negatively affected aphid fitness on GA-treated mycorrhizal plants 7-days post pea aphid feeding. Exogenous GA had a synergistic effect on JA- related plant gene expression in shoots, and increased JA and SA levels in leaves and iii petioles after 36 hours of aphid feeding. Pea aphid feeding for 36 hours induced SA- related defense gene expression and increased SA levels in leaves and petioles. In addition, root colonization by the AM fungus resulted in induced JA- and SA-related plant defense gene expression in shoots and roots, but decreased SA levels in leaves and petioles after 36 hours of aphid feeding. In objective 2, we found that GA signaling had an antagonistic effect on JA-related defense signaling after 36 hours of aphid feeding. In summary, this study serves as a foundation for future studies examining the role of GA in regulating plant defenses during insect-plant and insect-plant-AM fungus interactions. iv ACKNOWLEDGEMENTS I first and foremost want to thank my advisor, Dr. Karen Gomez for her amazing professional and personal support and commitment to graduate student success. Karen may not be the easiest on her students, but she expects the best from us, therefore bringing out the most of us. I have grown immensely over the past two years and could not have become who I am today without Karen pushing me to learn and grow. Although there were many tough times and long weeks, she was always there to lend out a helping hand. I have faced many challenges during my time at the University of Northern Colorado, however I was able to persevere through these because of Karen’s guidance and support. With that being said, thank you for everything! I would like to thank my committee members, Dr. Mitchell McGlaughlin and Dr. Hua Zhao for their patience, commitment, and guidance through this process. In addition, I could not have succeeded without the help of Dr. Cris Argueso and her Ph.D. candidate, Hannah Berry. Thank you, Cris for allowing me to use lab space and help fund the phytohormone analyses for my research. Thank you, Hannah Berry for taking the time to help me every step of the way through the phytohormone extraction and analyses. I cannot thank either of you two enough for being critical members of my success and I hope that I will be able to return the favor later down the road! I also could not have made it this far without the help of Dr. Jamie Riggs for helping me figure out which statistical analyses to use. I would have lost much more sleep over statistical analyses had she not provided me with her valuable assistance! v I want to thank one of my best friends, soon-to-be Dr. Tyler Sherman. Tyler has helped me through everything since the beginning of my time here at the University of Northern Colorado. Without him, I would never have had the chance to travel home or elsewhere with my family. Moreover, he has provided me with support throughout the entirety of this process whether it be personal and emotional support or in the lab. I know that I would not be the person that I am proud to be today without sharing friendship with this amazing individual. I also want to thank Christine Williams for helping me through a dark time early on in my graduate school career as well as being a great friend. I am not sure where I would be at had you not allowed me to open up to you as my first friend. You were the first person that I felt that I could trust after making the long and lonely journey here to Colorado. In addition, I want to thank my most recent of close friends Christopher Gregory and Lewis Eaton. These two remind me that there is more to life than being a scientist and spending all of my time towards my graduate school success. I am not sure that I would have made it through the past 8 months or so without them pulling me out of the dens of the Gomez lab, the teaching assistant office, or my apartment. In summary, my friends make up more than a large part of my success, but more importantly my happiness during these strenuous times. I hope that I have in some way, shape, or form have provided you all something of personal value as you all have provided to me. I want to thank my parents, Joel and Kelly Olson, my brother, Jaron Olson, and my grandparents, Don and Karen Olson, for their amazing support through thick and thin. Even when I began to shut myself out to the world, you all made sure to reach out to me and remind me that all things work out in the end. vi I want to thank the entire faculty, staff, graduate students, and undergraduate students at the University of Northern Colorado that have helped me succeed as well as provide an amazing place to grow and thrive. In, addition I want to thank the College of Natural and Health Sciences and Graduate Student Association for their financial support. I want to thank David Shimokawa for his undying commitment to graduate student success and making sure that students get the funding they need for research and for travel. I know that my time here would have been a much greater challenge without the funding. David also commits himself to graduate student happiness and health by organizing many events on and off campus to get people together and connected, and for that I am forever grateful. Finally, I want to thank The Plant Cell journal for providing me permission for the non-profit use of the figure provided in Fig. 1.4. vii TABLE OF CONTENTS CHAPTER I: INTRODUCTION AND REVIEW OF LITERATURE .............................. 1 Aims ........................................................................................................................ 1 Plant Interactions with Soil Beneficial Microbes: Arbuscular Mycorrhizal Symbioses ............................................................................................................... 3 Establishment of the Arbuscular Mycorrhizal Symbiosis ...................................... 4 Plant Interactions with Phloem-Feeding Insects ..................................................... 8 The Role of Phytohormones in Plant Biotic Interactions ..................................... 10 Jasmonic Acid Signaling and Plant Defenses ....................................................... 10 Salicylic Acid Signaling and Plant Defenses .......................................................
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