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A SEARCH for SOURCES of NONPERSISTENT VIRUS VECTORS and RESERVOIRS at LOCAL and REGIONAL SCALES Gina Marie Angelella Purdue University

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A SEARCH for SOURCES of NONPERSISTENT VIRUS VECTORS and RESERVOIRS at LOCAL and REGIONAL SCALES Gina Marie Angelella Purdue University Purdue University Purdue e-Pubs Open Access Dissertations Theses and Dissertations January 2015 TRACKING PLANT VIRUS INFECTIONS THROUGH MULTIPLE DIMENSIONS: A SEARCH FOR SOURCES OF NONPERSISTENT VIRUS VECTORS AND RESERVOIRS AT LOCAL AND REGIONAL SCALES Gina Marie Angelella Purdue University Follow this and additional works at: https://docs.lib.purdue.edu/open_access_dissertations Recommended Citation Angelella, Gina Marie, "TRACKING PLANT VIRUS INFECTIONS THROUGH MULTIPLE DIMENSIONS: A SEARCH FOR SOURCES OF NONPERSISTENT VIRUS VECTORS AND RESERVOIRS AT LOCAL AND REGIONAL SCALES" (2015). Open Access Dissertations. 1084. https://docs.lib.purdue.edu/open_access_dissertations/1084 This document has been made available through Purdue e-Pubs, a service of the Purdue University Libraries. Please contact [email protected] for additional information. Graduate School Form 30 Updated 1/15/2015 PURDUE UNIVERSITY GRADUATE SCHOOL Thesis/Dissertation Acceptance This is to certify that the thesis/dissertation prepared By Gina M. Angelella Entitled TRACKING PLANT VIRUS INFECTIONS THROUGH MULTIPLE DIMENSIONS: A SEARCH FOR SOURCES OF NONPERSISTENT VIRUS VECTORS AND RESERVOIRS AT LOCAL AND REGIONAL SCALES For the degree of Doctor of Philosophy Is approved by the final examining committee: Ian Kaplan Chair Jeffrey Holland Christian Krupke Joseph Anderson To the best of my knowledge and as understood by the student in the Thesis/Dissertation Agreement, Publication Delay, and Certification Disclaimer (Graduate School Form 32), this thesis/dissertation adheres to the provisions of Purdue University’s “Policy of Integrity in Research” and the use of copyright material. Approved by Major Professor(s): Ian Kaplan Steve Yaninek 12/9/2015 Approved by: Head of the Departmental Graduate Program Date TRACKING PLANT VIRUS INFECTIONS THROUGH MULTIPLE DIMENSIONS: A SEARCH FOR SOURCES OF NONPERSISTENT VIRUS VECTORS AND RESERVOIRS AT LOCAL AND REGIONAL SCALES A Dissertation Submitted to the Faculty of Purdue University by Gina M. Angelella In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy December 2015 Purdue University West Lafayette, Indiana ii For little Nadia iii ACKNOWLEDGEMENTS I would like to thank J. Anderson, C. Krupke, and J. Holland for their guidance and helpful comments regarding experimental design and chapter edits. C. Williams, J. Namachek, and D. Egel also collaborated on Chapter 1, V. Nalam, P. Nachappa, and J. White on Chapter 4, and A. Michel on Chapter 3. I would also like to thank the numerous extension agents, private pumpkin and alfalfa growers, and the staff at Purdue University’s Meigs Farm who made specimen collections possible. C. Blubaugh, J. Carillo, M. Garvey, L. Ingwell, E. Long, P. Olaya, and E. Rowen gave helpful comments for chapter edits. Chapter 4 is dedicated to the memory of Peter Saya, whose EPG expertise was instrumental in initiating experimental assays. C. Williams, J. Nemachek, and D. Egel collaborated on Chapter 1, V. Nalam, P. Nachappa, and J. White collaborated on Chapter 4, and A. Michel on Chapter 3. J. White and A. Dehnel provided Aphis craccivora colonies and rearing expertise, D. Lagos and D. Voegtlin provided aphid identification expertise and training, S. Pearce, D. Egel, and S. Hoke assisted with sample collection, C. Michel with RNA extraction, S. Nouri and M. Deb provided advice regarding RNA extraction and/or primer design, B. Peterson assisted with nucleic acid extraction techniques, and J.L. White provided technical assistance with LINUX and STACKS installation. Funding support was provided by the USDA North Central Region IPM Grants Program, grant 11-34103-30723, the Indiana Vegetable Growers Association Grant Program, the Indiana Academy of Sciences Professional Grant Program, and NIFA USDA Predoctoral Fellowship grant 107483. iv TABLE OF CONTENTS Page ABSTRACT .............................................................................................................................. vi CHAPTER 1 ............................................................................................................................... 1 Introduction ........................................................................................................................ 1 Methods .............................................................................................................................. 3 Results ................................................................................................................................ 7 Discussion ......................................................................................................................... 17 References......................................................................................................................... 22 CHAPTER 2 ............................................................................................................................. 28 Introduction ...................................................................................................................... 28 Methods ............................................................................................................................ 29 Results .............................................................................................................................. 32 Discussion ......................................................................................................................... 35 References......................................................................................................................... 39 CHAPTER 3 ............................................................................................................................. 43 Introduction ...................................................................................................................... 43 v Page Methods ............................................................................................................................ 44 Results .............................................................................................................................. 47 Discussion ......................................................................................................................... 52 References......................................................................................................................... 55 CHAPTER 4 ............................................................................................................................. 58 Introduction ...................................................................................................................... 58 Methods ............................................................................................................................ 60 Results & Discussion ......................................................................................................... 62 References......................................................................................................................... 67 APPENDICES Appendix A ....................................................................................................................... 70 Appendix B ....................................................................................................................... 91 Appendix C ....................................................................................................................... 96 VITA ...................................................................................................................................... 106 vi ABSTRACT Angelella, Gina M. Ph.D., Purdue University, December 2015. Tracking Plant Virus Infections Through Multiple Dimensions: A Search for Sources of Nonpersistent Virus Vectors and Reservoirs at Local and Regional Scales. Major Professor: Ian Kaplan. My dissertation explores the ecology of aphid-vectored viruses in Midwestern cucurbits; in particular, it focuses on identifying source populations of vectors and virus reservoirs within fields and interactions of vector aphids with land cover surrounding and within fields. Initially, I identified the most commonly occurring viruses and aphid species associated with virus infections in pumpkin fields located across Indiana. This was done by assaying cucurbit leaf tissue with multiplex-rt-PCR targeting all four aphid-vectored, nonpersistent viruses found in cucurbits (papaya ringspot virus type-W, watermelon mosaic virus type-2, zucchini yellow mosaic virus, cucumber mosaic virus) and concurrently monitoring aphid species alightment in fields throughout Indiana. Watermelon mosaic virus (WMV) was the most common infection, detected in all but one field across both years. Papaya ringspot virus (PRSV) was also detected in many fields, but not zucchini mosaic virus or cucumber mosaic virus. Therioaphis trifolii and Aphis craccivora were positively associated with PRSV, and Aphis nerii with WMV. Rhopalosiphum padi was negatively associated with WMV. No relationship between total aphid or noncolonizing species counts was found with virus infection, and an increase in total colonizing species (Aphis gossypii) trapped in fields predicted a decrease in PRSV infection. Negative relationships between aphid species alightment and virus infection in fields could result from a virus-induced reduction in pumpkin plant quality, thereby reducing abundance of a colonizing species within the field. Other virus-mediated
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