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Incidence of Xylella Fastidiosa in Oklahoma, a Survey of Potential Insect Vectors, and Identification of Potential Plant Reservoir Hosts

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Incidence of Xylella Fastidiosa in Oklahoma, a Survey of Potential Insect Vectors, and Identification of Potential Plant Reservoir Hosts INCIDENCE OF XYLELLA FASTIDIOSA IN OKLAHOMA, A SURVEY OF POTENTIAL INSECT VECTORS, AND IDENTIFICATION OF POTENTIAL PLANT RESERVOIR HOSTS By LISA MARIE OVERALL Bachelor of Science in Biology University of Central Oklahoma Edmond, OK 2005 Master of Science in Entomology and Plant Pathology Oklahoma State University Stillwater, OK 2008 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY May, 2013 INCIDENCE OF XYLELLA FASTIDIOSA IN OKLAHOMA, A SURVEY OF POTENTIAL INSECT VECTORS, AND IDENTIFICATION OF POTENTIAL PLANT RESERVOIR HOSTS Dissertation Approved: Dr. Eric Rebek Dissertation Adviser Dr. Phil Mulder Dr. Astri Wayadande Dr. Udaya DeSilva ii Name: LISA MARIE OVERALL Date of Degree: MAY, 2013 Title of Study: INCIDENCE OF XYLELLA FASTIDIOSA IN OKLAHOMA, SURVEY OF POTENTIAL INSECT VECTORS, AND IDENTIFICATION OF POTENTIAL PLANT RESERVOIR HOSTS Major Field: ENTOMOLOGY Abstract: The bacterium Xylella fastidiosa is the causative agent of specific plant diseases associated with peach, plum, oleander, almond, elm, oak, pecan, sycamore, and Pierce’s disease of grapes. The bacterium is transmitted by xylem sap-feeding insects. In recent years, Pierce’s disease was detected in eight counties in central and northeastern Oklahoma, prompting further investigation of the disease epidemiology in this state. I surveyed Oklahoma for xylem sap-feeding insects to determine the species composition within vineyards, nurseries, and pecan orchards and whether or not they harbored X. fastidiosa. A second objective involved determining the natural inoculativity of insect vectors commonly occurring in vineyards throughout the state. In the third objective, I surveyed common weeds in vineyards to identify potential reservoir hosts of X. fastidiosa. Weedy plants sampled in this study are known or suspected food sources of xylem sap-feeding insect vectors, are commonly found in Oklahoma vineyards, and have been implicated as potential reservoirs for X. fastidiosa. Immunocapture polymerase chain reaction was used to test all insect and plant samples in this study. I found the following sharpshooters in my survey: Xyphon flaviceps, Draeculacephala navicula, D. robinsoni, Graphocephala coccinea, G. versuta, G. hieroglyphica, Cuerna costalis, Oncometopia orbona, Homalodisca vitripennis, Prosapia bicincta, and Clastoptera xanthocephala. The three most frequently captured species in vineyards and tree nurseries included G. versuta, C. xanthocephala, and O. orbona. From a subsample of insects screened for X. fastidiosa, 2.4% tested positive for the bacterium. Field-collected G. versuta inoculated X. fastidiosa to susceptible indicator plants, ragweed and alfalfa. A higher percentage of alfalfa became infected than ragweed following inoculation periods. Very few plants tested positive and none of the plants testing positive are considered new hosts of X. fastidiosa. These results indicate that weedy reservoir hosts may not be a contributing factor to the epidemiology of Pierce’s disease in Oklahoma. From this study, I found that insect vectors were present and persistently captured from vineyards, nurseries and pecan orchards. The most predominant insect vector, G. versuta, was most abundant in June and July and was infective and competent to spread X. fastidiosa to susceptible plant host. iii ACKNOWLEDGEMENTS I thank my Advisor, Dr. Eric Rebek, for his guidance, support, and willingness to encourage me always. He was not only my advisor, but was a mentor and a friend. He encouraged a positive attitude and kept me motivated when I needed it most. To him I am thankful. I would like to thank my committee members, Drs. Phil Mulder, Astri Wayadande, and Udaya De Silva. Dr. Mulder has provided me unwaivering support throughout my education at Oklahoma State University. Dr. Wayadande provided me laboratory access and was always willing to share her experience and knowledge. Dr. DeSilva provided invaluable advice regarding my molecular methods. Dr. Damon Smith was initially on my committee and helped me to develop my molecular protocols. I thank him. I extend my gratitude to the Plant Disease and Insect Diagnostic Lab (PDIDL), Oklahoma State University. Jen Olson guided me through my protocols, allowed me use of clean bench space, and was a friend. To her and the PDIDL lab, I am thankful. I thank the faculty and staff in the Department of Entomology and Plant Pathology, Oklahoma State University. My funding was provided by the Department of Entomology and Plant Pathology and Oklahoma Department of Commerce, Viticulture and Enology Development Fund. Thank you. iv Acknowledgements reflect the views of the author and are not endorsed by committee members or Oklahoma State University. There were many students that assisted me with my research including: Mark Lovelace, Derek Barchenger, Bailey Adams, Colby Gregg, Jackson Seibert, and Ashley Harris. Finally, I must thank my Grandma for always believing in me and teaching me to enjoy life. I thank my Father, Coy Overall, for his support and friendship throughout my graduate education. I thank Tres Harriman for his love, support, and friendship. v Acknowledgements reflect the views of the author and are not endorsed by committee members or Oklahoma State University. TABLE OF CONTENTS Chapter Page I. INTRODUCTION ......................................................................................................1 Objectives ................................................................................................................4 Literature Cited ........................................................................................................5 II. LITERATURE REVIEW ..........................................................................................9 Plant Colonization ..................................................................................................10 Bacterial Colonization Insect Foregut ...................................................................12 Insect Inoculation ...................................................................................................13 Plant Host Entry and Infection ...............................................................................14 Cell-Cell Signaling and Biofilm Formation ...........................................................16 Differentiation of Subspecies of Xylella fastidiosa ...............................................19 Background of Diseases Caused by Xylella fastidiosa ..........................................22 Insect Vectors and Their Biology ..........................................................................33 Literature Cited ......................................................................................................45 III. DISTRIBUTION, PEAK ABUNDANCE, AND INFECTION STATUS OF POTENTIAL INSECT VECTORS OF XYLELLA FASTIDIOSA, THE CAUSAL AGENT OF PIERCE’S DISEASE OF GRAPES AND BACTERIAL LEAF SCORCH OF SHADE TREES IN OKLAHOMA ................................................66 Abstract ..................................................................................................................66 Introduction ............................................................................................................67 Materials and Methods ...........................................................................................70 Results and Discussion ..........................................................................................75 Conclusions ............................................................................................................85 Literature Cited ......................................................................................................91 vi Chapter Page IV. THE NATURAL INOCULATIVITY OF INSECT VECTORS OF XYLELLA FASTIDIOSA COLLECTED FROM OKLAHOMA VINEYARDS ..................113 Abstract ................................................................................................................113 Introduction ..........................................................................................................114 Materials and Methods .........................................................................................117 Results and Discussion ........................................................................................121 Conclusions ..........................................................................................................125 Literature Cited ....................................................................................................126 V. IDENTIFICATION OF WEEDY PLANTS ASSOCIATED WITH OKLAHOMA VINEYARDS THAT COULD SERVE AS SOURCES OF XYLELLA FASTIDIOSA........................................................................................................139 Abstracts ..............................................................................................................139 Introduction ..........................................................................................................140 Materials and Methods .........................................................................................142 Results and Discussion ........................................................................................147 Conclusions ..........................................................................................................149 Literature Cited ...................................................................................................150
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