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Abstract Jacobson, Alana Lynn ABSTRACT JACOBSON, ALANA LYNN. Tomato spotted wilt virus (TSWV) in North Carolina: Characterizing the Vector Competence of Thrips tabaci and Investigating the Use of Cyantraniliprole for Reducing Transmission of TSWV by Frankliniella fusca and Frankliniella occidentalis. (Under the direction of George G. Kennedy.) Tomato spotted wilt virus (TSWV), is a thrips transmitted plant virus that ranks among the most economically important insect-vectored plant viruses worldwide. In the Southeastern U.S. this virus causes economic losses in many crops, including tobacco, pepper, tomato, and peanut. In North Carolina the two primary thrips vectors implicated in primary and secondary spread of TSWV, respectively, are Frankliniella fusca Hinds and F. occidentalis Pergande. The role of the third vector species, Thrips tabaci Lindeman, in the epidemiology of TSWV in NC is unknown. T. tabaci has not been thought to contribute significantly to virus spread due to its localized importance as a vector in other parts of the world and documented variation in vector competence that exists among different populations. T. tabaci populations that efficiently transmit TSWV have been observed in the U.S.; however, the extent of variation in transmission efficiency that exists among NC populations is unknown. To better characterize the competence of NC populations of T. tabaci to transmit TSWV, T. tabaci collected at multiple locations in NC during 2010 were tested for their ability to transmit multiple TSWV isolates that were collected from the same locations. Results showed that vector competence of T. tabaci isofemale lines varied among TSWV isolates in a manner that was isolate specific. Moreover, transmissibility of isolates varied in a manner that was specific to T. tabaci isofemale line. On average transmission rates of the TSWV isolates were higher when they were transmitted by thrips collected from the same location, which suggests local adaptation between the thrips and the TSWV isolates. To better understand the role of the virus and thrips in this observed variation, a population genetic study of thrips populations was conducted, which included individuals from T. tabaci isofemale lines used in the transmission study. Microsatellite and mitochondrial COI DNA markers were used to examine population structuring in T. tabaci across NC, and the variation in TSWV transmission phenotypes among different clonal groups of T. tabaci. Geographic structuring of T. tabaci occurred across the locations sampled, and the distribution of clonal groups suggests that dispersal occurs among three of the four locations where thrips populations were collected. In addition, analysis of the probability of TSWV transmission by T. tabaci in relation to a specific ‘clonal assignment’ of individual thrips, based on microsatellite allele frequencies, suggests that the interaction of specific thrips genotypes with specific virus isolates is a better predictor of of TSWV transmission by T. tabaci than sympatry. In a separate study, the potential of cyantraniliprole, an anthranilic diamide insecticide, for reducing the spread of TSWV by F. fusca and F. occidentalis was investigated in a greenhouse study. Transmission of TSWV by F. fusca to Capsicum annuum L. seedlings was reduced in plants treated with Cyazypyr™, but transmission of TSWV by F. occidentalis was not. Mortality of F. fusca at 3 days post treatment did not differ significantly on excised foliage of cyantraniliprole treated and control plants, but feeding injury was significantly less on treated foliage. Overall, results indicate that T. tabaci has the potential to contribute to TSWV spread in NC; however, the role of T. tabaci in the epidemiology of TSWV can be expected to vary depending on the specific vector -virus isolate combinations that are present. In addition, cyantraniliprole may be a useful tool for TSWV management programs, and further studies investigating its potential to decrease TSWV incidence are warranted. © Copyright 2011 by Alana Lynn Jacobson All Rights Reserved Tomato spotted wilt virus (TSWV) in North Carolina: Characterizing the Vector Competence of Thrips tabaci and Investigating the Use of Cyantraniliprole for Reducing Transmission of TSWV by Frankliniella fusca and Frankliniella occidentalis by Alana Lynn Jacobson A dissertation submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the Degree of Doctor of Philosophy Entomology Raleigh, North Carolina 2012 APPROVED BY: ___________________________________ _________________________________ George G. Kennedy Hannah J. Burrack Committee Chair ___________________________________ _________________________________ Edward Vargo Christopher Gunter BIOGRAPHY Thrips, ‘tiny slender insects with fringed wings’, were the center of Alana Jacobson’s focus while pursuing a Ph.D. researching onion thrips’ role in the epidemiology of a Tomato spotted wilt virus in North Carolina. Born and raised in Phoenix, Arizona, she developed an interest in entomology and insect pest management in an agricultural biotechnology class and through her participation in the FFA Entomology Contest during high school. The experiences she gained through these programs fueled her desire to continue her studies at New Mexico State University where she received a B.S in Agricultural Biology and a B.A. in Spanish. In addition to class work, she was employed in departmental laboratories, gained valuable research experience in nematology and entomology, and participated in a year-long study abroad program in Mexico. In 2005 she moved to West Point, IN and spent the next three years completing a M.S. in Entomology at Purdue University investigating insecticide resistance in the corn earworm. The next leg of Alana’s eastward journey brought her to her current position here at NC State working under the direction of George Kennedy on insect vectored plant diseases. Upon completion of her Ph.D. she looks forward to entering a career researching and solving contemporary pest management problems. In her free time she enjoys horseback riding, hunting, quilting, and looking for cool rocks. ii ACKNOWLEDGMENTS The past four years have been great, mainly because of the wonderful people who have been part of my life, and who have helped me through this challenging and exciting time. First, I would like to thank my advisor, George Kennedy, for giving me the opportunity to work on this exciting and diverse project. You are an exemplary mentor, a good friend, and I have learned so much working with you on this project. Input from my committee members, Hannah Burrack, Ed Vargo, and Chris Gunter, has also been invaluable to the success of this project and enabled me to apply for, and attain postdoctoral funding. Dr. Gunter, thank you for your input and encouragement throughout this entire project. Dr. Vargo, thank you for your advice and support, especially during the population genetic study. Dr. Burrack, thank you for your advice, encouragement, friendship, and many conversations, but most of all, thanks for introducing me to Wisconsin cheese curds. Next I would like to thank the numerous people around our lab and the department who have helped with various aspects of this project: Carol Berger for teaching me the ropes of thrips rearing, TSWV isolate maintenance, and for being the one person in the lab I could always talk into a doughnut or milkshake run; Damon D’Ambrosio for his invaluable help with thrips colony maintenance, for help conducting numerous research projects in the lab (there are too many to list), his quick wit and dry sense of humor; Dan “the thrips-ID man” Grist for his help with slide mounting and thrips identification; Amanda Beudoin and Shannon Morsello for helping me get started on my lab work and advising me on classes; Warren Booth for helping me with various aspects of conducting molecular work and analyzing data; Paul Labadie for his help with lab work during the population genetics study; Mark Abney for his help and insights, good conversation, and for giving me the opportunity to practice shooting moving targets; Fred Gould for his comments and feedback on my grant proposal; Joy Smith and Consuello Arellano for their help with statistical analyses; and most importantly, Gene Dupree, Pat Bachelor, Jean Carter and Joyce Taylor, the four people who were really running the department. iii I am also thankful for the many friends who have helped balance my life, and provided encouragement and pleasant distractions from the demands of grad school. A special thanks goes to Diane Silcox for her friendship, beagle-sitting, the numerous meals she cooked for me, and late-night help with thrips wrangling on several occasions; Kelly Oten for fiestas; and Eleanor Spicer for many successful days. I am also grateful to Linda and Jeff Mullen for giving me the opportunity to ride horses at their farm, and for all of the Carousel Farms Crew, especially Cindy Gammon, Mennette Price, Rhonda and Christian Chalmers, Brooke Biddle, Meg Tucker, Darby Hollenbach, Remy Metzger, Meghan Giles, Dana and Teresa Jones, Weldon Gammon, and Danny Young. Last but not least, thank you to my family and friends back home for their encouragement and support throughout my career as a student. I love you all. iv TABLE OF CONTENTS LIST OF TABLES ......................................................................................................... vi LIST OF FIGURES ......................................................................................................vii
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