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Host Plant Use by Insect Herbivores Mediated by Microorganisms

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Host Plant Use by Insect Herbivores Mediated by Microorganisms HOST PLANT USE BY INSECT HERBIVORES MEDIATED BY MICROORGANISMS A Dissertation by JOSEPHINE BEMA ANTWI Submitted to the Office of Graduate and Professional Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Chair of Committee, Raul F. Medina Co-Chair of Committee, Gregory A. Sword Committee Members, Micky Eubanks Mariana Mateos Jesus Esquivel Head of Department, David Ragsdale August 2015 Major Subject: Entomology Copyright 2015 Josephine B. Antwi ABSTRACT Geographic isolation plays a major role in population divergence, but host-plants can also generate selective forces that drive genetic differentiation. Host-plant associations of insects can also be mediated by their associations with microorganisms, some of which are linked to pest status on crops. However, the ecological and evolutionary roles these microorganisms play in plant-insect interactions are not fully understood, neither are their implications for pest management. My dissertation applied an interdisciplinary approach to explore the potential role of microorganisms in plant-insect interactions. Two important agricultural pests, cotton fleahopper (CFH) (Pseudatomoscelis seriatus), a cotton pest, and sugarcane aphid (Melanaphis sacchari), an invasive pest of sorghum, were used as model organisms in my studies. Using amplified fragment length polymorphisms (AFLPs) I sampled CFH from 12 host-plants. I found that CFH genotypes are distinct on only 2 out of 12 uncultivated plant species. My results suggest that several uncultivated hosts likely constitute a source of CFH for cotton. To determine whether bacterial symbionts correlated with genetic variation in CFH host-association, I compared bacterial communities within CFH using 454 pyrosequencing of the 16S rRNA gene. There was no correlation between the two. Although CFH is widely distributed across cotton-growing regions in the U.S., it is considered a major cotton pest only in certain regions while in others it is rarely a pest. I compared bacterial communities within CFH using 16S rRNA pyrosequencing to test the potential for bacterial symbionts to influence variation in CFH pest status. Results from this ii bacterial survey failed to find any correlation between variation in pest status of CFH and bacterial community composition. I also explored the roles of potentially beneficial symbiotic fungi (i.e., fungal endophytes) in improving plant development and providing resistance against herbivory. Soaking sorghum seeds in liquid suspensions of two fungal endophytes increased seedling height and fresh biomass. Additionally, I detected that endophyte treatment significantly affected the reproduction and behavior of sugarcane aphids, but performance was conditional on the identity of the endophyte and the part of the plant where aphids were located. Overall, findings from my studies improve our understanding of the ecological and evolutionary implications of plant-microbe-insect interactions. iii DEDICATION To my mother, Margaret Antwi, I would not have accomplished this if it hadn’t been for your support and encouragement. To my brothers, Alfred Antwi and Richard Owusu Banahene and my sister, Christina Tawiah, for their unfailing encouragement and love. iv ACKNOWLEDGEMENTS I would like to express my deep appreciation and gratitude to my PhD advisor, Raul Medina and co-advisor, Greg Sword. Throughout my PhD training they have provided me with the guidance and mentorship that have prepared me both intellectually and socially towards my career goals. Their commitment and hard work to research and the to the well- being of their students have been an inspiration to me. I am indebted to my committee members: Micky Eubanks, Mariana Mateos and Jesus Esquivel for their inputs and insightful comments to improve my dissertation research. They spent countless hours, during both our committee meetings and through personal conversations, providing constructive criticisms that have collectively advanced my thinking as a researcher. I am also grateful to the faculty at the Entomology Department especially Robert Wharton, Jeff Tomberlin, Kevin Heinz and Spencer Behmer for their insightful comments about my research and mentoring me as a teaching assistant during my PhD training at the Department. I would like to thank my collaborators, Mike Brewer (Entomology Department, TAMU), Peter Ellsworth (University of Arizona), Bill Rooney (Department of Soil and Crop Sciences, TAMU) and Apurba Barman for providing me with insect and plant samples for my research. I thank Dale Kruse (Tracy Herbarium, TAMU) for his assistance identifying plants used in my study. I would also like to thank the Texas EcoLab program for providing funding and research sites for my studies. I especially thank the landowners for giving me access to their properties to conduct my research. I am grateful to Cesar Valencia, Dr. Julissa Ek-Ramos and v Dr. Vijay Verma for their assistance during lab experiments and field work. I thank my lab mates: Kyle Harrison, Suhas Vyavhare, Diana Castillo-Lopez, Carrie Deans, Lauren Kalns and Wenqing Zhou for their support and assistance in the lab. I also thank all my friends, especially Claudia Castillo-Gonzalez and Diana Solis and many others who have been supportive in various aspects of my PhD training. vi TABLE OF CONTENTS Page ABSTRACT ....................................................................................................................... ii DEDICATION .................................................................................................................. iv ACKNOWLEDGEMENTS ............................................................................................... v TABLE OF CONTENTS ................................................................................................. vii LIST OF FIGURES .......................................................................................................... ix LIST OF TABLES ............................................................................................................. x CHAPTER I INTRODUCTION ........................................................................................................ 1 II HOST-ASSOCIATED DIFFERENTIATION IN A HIGHLY POLYPHAGOUS, SEXUALLY REPRODUCING INSECT HERBIVORE ............................................... 7 Synopsis ...................................................................................................................... 7 Introduction ................................................................................................................ 8 Materials and Methods ............................................................................................. 11 Results ...................................................................................................................... 18 Discussion ................................................................................................................ 26 III LACK OF CORRELATION BETWEEN HOST-ASSOCIATED DIFFERENTIATION (HAD) AND MICROBIOME OF COTTON FLEAHOPPER, PSEUDATOMOSCELIS SERIATUS (HEMIPTERA: MIRIDAE) ............................... 30 Synopsis .................................................................................................................... 30 Introduction .............................................................................................................. 31 Materials and Methods ............................................................................................. 34 Results ...................................................................................................................... 39 Discussions ............................................................................................................... 50 IV VARIATION IN MICROBIOME DOES NOT EXPLAIN THE PEST STATUS OF THE COTTON FLEAHOPPER ................................................................................... 54 Synopsis .................................................................................................................... 55 Introduction .............................................................................................................. 56 Materials and Methods ............................................................................................. 61 Results ...................................................................................................................... 71 vii Discussion ................................................................................................................ 78 CHAPTER V FUNGAL ENDOPHYTES, BEAUVERIA BASSIANA AND PURPUREOCILIUM LILACINUM, POSITIVELY AFFECT THE GROWTH OF SORGHUM AND IMPACT THE BEHAVIOR AND PERFORMANCE OF THE SUGARCANE APHID, MELANAPHIS SACCHARI .......................................................................................... 84 Synopsis .................................................................................................................... 84 Introduction .............................................................................................................. 85 Materials and Methods ............................................................................................. 88 Results ...................................................................................................................... 93 Discussion ...............................................................................................................
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