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USING NEW MOLECULAR TOOLS to EXPLORE HOW ORGANIC FARMING IMPACTS TOP-DOWN and BOTTOM-UP REGULATION of POTATO HERBIVORES by KARO

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USING NEW MOLECULAR TOOLS to EXPLORE HOW ORGANIC FARMING IMPACTS TOP-DOWN and BOTTOM-UP REGULATION of POTATO HERBIVORES by KARO USING NEW MOLECULAR TOOLS TO EXPLORE HOW ORGANIC FARMING IMPACTS TOP-DOWN AND BOTTOM-UP REGULATION OF POTATO HERBIVORES By KAROL LYNN KREY A dissertation submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY WASHINGTON STATE UNIVERSITY Department of Entomology JULY 2017 © Copyright by KAROL LYNN KREY, 2017 All Rights Reserved © Copyright by KAROL LYNN KREY, 2017 All Rights Reserved To the Faculty of Washington State University: The members of the Committee appointed to examine the dissertation of KAROL LYNN KREY find it satisfactory and recommend that it be accepted. William E. Snyder, Ph.D., Chair David W. Crowder, Ph.D. John P. Reganold, Ph.D. Paul D. Nabity, Ph.D. ii ACKNOWLEDGMENT I would like to thank Bill Snyder, my advisor and committee chair, for the opportunity to be a part of the lab and for his help throughout my graduate career. I have made leaps and bounds in progress toward becoming a great researcher, writer, and mentor. I would also like to thank my committee members Dave Crowder, John Reganold, and Paul Nabity, for providing me guidance and constructive criticisms on my dissertation projects. I am grateful for the many friends and comrades I have made in the entomology department and hope to always stay in touch. I am lucky to have had the support from the Snyder lab team, especially, Christine Lynch, Amanda Meadows, Carmen Castillo, Jake Asplund, Matt Jones, Joseph Taylor, Olivia Smith, and the super amazing post-docs, Daisy Fu and Carmen Blubaugh (your help and support was invaluable). Thanks to all the undergraduate workers that spent tireless hours helping me: Abbey Estep, Ashley Norberg, Trevor Snodgrass, Jen Madigan, and Samantha Beck. Last, but most certainly not least, I would like to thank my family for their constant encouragement throughout my dissertation work. They were always there with positive words of advice to help me achieve my goals. I am especially indebted to my husband, Jesse Burke, whose unwavering love and support made everything possible. iii USING NEW MOLECULAR TOOLS TO EXPLORE HOW ORGANIC FARMING IMPACTS TOP-DOWN AND BOTTOM-UP REGULATION OF POTATO HERBIVORES Abstract by Karol Lynn Krey, Ph.D. Washington State University July 2017 Chair: William E. Snyder Herbivorous agricultural pests are simultaneously threatened by predatory natural enemies and by toxic plant defenses. Both these “top down” and “bottom up” forces might be enhanced under organic farm management. For example, reductions in the use of broad-acting insecticides, typical of some organic systems, can lead to dramatically higher predator density and biodiversity on organic compared to conventional farms. Similarly, efforts by organic farmers to increase soil organic matter might lead to more robust plants better able to activate effective anti-herbivore defenses. We took advantage of modern molecular approaches to track predation and quantify the deployment of plant defenses for potatoes grown under organic or conventional practices in eastern Washington, USA. Chapter 2 reports significantly higher densities of predatory Geocoris and Nabis bugs in organic potato fields; because per-capita predation rates on spider mites were generally similar in the two farming systems, we would expect stronger top-down suppression of spider mites in organic than conventional potato fields. Chapter 3 reports greater defense-gene activity for potato foliage on organic than conventional fields of the potato variety Norkotah, although no such effect was seen for the variety Alturas. Chapter 4 reports heightened survivorship of Colorado potato beetles in the greenhouse on potato iv plants grown in organic than conventional soils, but was unaffected by the presence of aphids. In contrast, aphids reached significantly higher densities when reared alone than when paired with Colorado potato beetles, and were unaffected by soil type. Across the field and greenhouse studies reported in Chapters 3 and 4, we saw weakly heightened soil microbial activity in organic compared to conventional soils, but few differences in plant chemistry. Thus, further work is needed to explore the ecological basis of differences in plant defenses or herbivore survivorship mediated by soil management practices. Altogether, the work described in my dissertation suggests that the potential to alter and exploit organic versus conventional management practices to maximize plant deployment of natural defenses could form an exciting new approach to sustainable agriculture and host-plant resistance. v TABLE OF CONTENTS Page ACKNOWLEDGMENTS ........................................................................................................ iii ABSTRACT ............................................................................................................................... iv LIST OF TABLES ..................................................................................................................... ix LIST OF FIGURES .................................................................................................................... x CHAPTER CHAPTER ONE: INTRODUCTION ............................................................................. 1 Herbivore suppression from above and below .......................................................... 1 Top-down suppression of herbivores in agricultural systems................................... 3 Plant defense against herbivores – Evolution and types of defensive traits ............. 4 Plant defense costs and benefits ................................................................................ 5 Plant compounds related to defense .......................................................................... 7 Direct induced defense genes.................................................................................... 8 Soil resources related to plant health and defense against herbivory ....................... 9 Organic farming’s impact on top-down and bottom-up processes in potato crops 11 Dissertation overview ............................................................................................. 13 References ............................................................................................................... 15 CHAPTER TWO: COULD GENERALIST PREDATORS DIFFUSE SPIDER MITE OUTBREAKS ON ORGANIC FARMS? .................................................................... 37 ABSTRACT ............................................................................................................ 37 1. INTRODUCTION .............................................................................................. 38 2. MATERIALS AND METHODS ........................................................................ 41 vi Primer design ................................................................................................... 41 DNA detectability feeding trials ...................................................................... 42 Arthropod survey and collections in commercial potato fields ....................... 43 Molecular gut-content analysis ....................................................................... 45 Statistical analyses ........................................................................................... 45 3. RESULTS ........................................................................................................... 46 Two-spotted spider mite primers ..................................................................... 46 Gut retention times of spider mites in predator guts ....................................... 47 Predator and pest densities in commercial potato fields ................................. 48 Detection of prey DNA in predators ............................................................... 48 4. DISCUSSION ..................................................................................................... 49 5. REFERENCES ................................................................................................... 54 CHAPTER THREE: CAN ORGANIC FARMING SHARPEN PLANT DEFENSES AGAINST HERBIVORY? ........................................................................................... 72 ABSTRACT ............................................................................................................ 72 1. INTRODUCTION .............................................................................................. 73 2. MATERIALS AND METHODS ........................................................................ 76 Study Area ....................................................................................................... 76 Quantifying Gene-Expression Patterns in Potato ............................................ 76 Arthropod Sampling ........................................................................................ 78 Analysis of Soil Properties .............................................................................. 79 Analysis of Foliar Metabolites and Nutrients .................................................. 81 Statistical analyses ........................................................................................... 82 vii 3. RESULTS ........................................................................................................... 85 Plant transcriptomics and differentially expressed genes ................................ 86 Pest densities in potato fields .......................................................................... 87 Soil nutrients ...................................................................................................
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