2019 Research Progress Reports Research Progress Reports Pierce’s Disease and Other Designated Pests and Diseases of Winegrapes - December 2019 - Compiled by: Pierce’s Disease Control Program California Department of Food and Agriculture Sacramento, CA 95814 2019 Research Progress Reports Editor: Thomas Esser, CDFA Cover Design: Sean Veling, CDFA Cover Photograph: Photo by David Köhler on Unsplash Cite as: Research Progress Reports: Pierce’s Disease and Other Designated Pests and Diseases of Winegrapes. December 2019. California Department of Food and Agriculture, Sacramento, CA. Available on the Internet at: https://www.cdfa.ca.gov/pdcp/Research.html Acknowledgements: Many thanks to the scientists and cooperators conducting research on Pierce’s disease and other pests and diseases of winegrapes for submitting reports for inclusion in this document. Note to Readers: The reports in this document have not been peer reviewed. 2019 Research Progress Reports TABLE OF CONTENTS Section 1: Xylella fastidiosa and Pierce’s Disease REPORTS • Addressing Knowledge Gaps in Pierce’s Disease Epidemiology: Underappreciated Vectors, Genotypes, and Patterns of Spread Rodrigo P.P. Almeida, Monica L. Cooper, Matt Daugherty, and Rhonda Smith ......................2 • Testing of Grapevines Designed to Block Vector Transmission of Xylella fastidiosa Rodrigo P.P. Almeida ..............................................................................................................11 • Field-Testing Transgenic Grapevine Rootstocks Expressing Chimeric Antimicrobial Protein and Polygalacturonase Inhibitory Protein Abhaya M. Dandekar ...............................................................................................................15 • Field Evaluation of Cross-Graft Protection Effective Against Pierce's Disease by Dual DNA Constructs Expressed in Transgenic Grape Rootstocks David Gilchrist .........................................................................................................................24 • Transgenic Rootstock-Mediated Protection of Grapevine Scion by Introduced Single and Dual DNA Constructs David Gilchrist, James Lincoln, Abhaya M. Dandekar, David Tricoli, and Bryan Pellissier ...................................................................................................................................31 • Biological Control of Pierce's Disease of Grape with an Endophytic Bacterium Steven Lindow .........................................................................................................................41 • Optimizing Biological Control of Pierce's Disease with Paraburkholderia phytofirmans Steven Lindow .........................................................................................................................57 • Geographic Distribution of Isolate Virulence in Xylella fastidiosa Collected from Grape in California and Its Effect on Host Resistance Rachel Naegele and Leonardo De La Fuente ..........................................................................66 • Characterization of the Lipopolysaccharide-Mediated Response to Xylella fastidiosa Infection in Grapevine Caroline Roper and Dario Cantu..............................................................................................73 • Generating Pierce’s Disease Resistant Grapevines Using CRISPR/Cas9 and Traditional Transgenic Approaches Caroline Roper and Dario Cantu..............................................................................................84 - i - 2019 Research Progress Reports • Grape Protoplast Isolation and Regeneration of Plants for Use in Gene Editing Technology David Tricoli ............................................................................................................................92 • Breeding Pierce’s Disease Resistant Winegrapes Andrew Walker ......................................................................................................................104 • Molecular Breeding Support for the Development of Pierce's Disease Resistant Winegrapes Andrew Walker and Dario Cantu ..........................................................................................116 ABSTRACTS • Cultivar Susceptibility and Temperature-Dependent Recovery of Xylella fastidiosa Infected Grapevines Lindsey Burbank ....................................................................................................................130 • Searching for Endosymbionts in Kolla paulula, a Vector of Xylella fastidiosa Causing Pierce’s Disease of Grapevine in Taiwan Jianchi Chen, Hsien-Tzung Shih, Chiou-Chu Su, and Chung-Jan Chang .............................131 • Insights for Understanding Nemaguard Resistance Mechanism Against Xylella fastidiosa Rodrigo Krugner, Craig Ledbetter, Elizabeth Rogers, and Lindsey Burbank .......................133 • Molecular Characterization and Functional Analysis of PILH in the Pathogenicity of Xylella fastidiosa Hong Lin ................................................................................................................................134 Section 2: Glassy-winged Sharpshooter REPORTS • Riverside County Glassy-winged Sharpshooter Program: Temecula Valley and Coachella Valley Matt Daugherty, Carmen Gispert, and Christina Hoddle ......................................................136 • Management of Insecticide Resistance in Glassy-winged Sharpshooter Populations Using Toxicological, Biochemical, and Genomic Tools Richard Redak, Bradley White, and Frank Byrne .................................................................141 - ii - 2019 Research Progress Reports Section 3: Other Pests and Diseases of Winegrapes REPORTS • Grapevine Virus Management in Lodi: A Collaborative Research and Integrated Outreach Effort to Help Solve a Statewide Challenge Stephanie L. Bolton ...............................................................................................................151 • A Study on the Impact of Individual and Mixed Leafroll Infections on the Metabolism of Ripening Winegrape Berries Dario Cantu, Maher Al Rwahnih, Susan Ebeler, and Deborah Golino, ...............................167 • Improving Extension Outcomes: Identifying Drivers and Barriers to Adoption of Management Practices Using Leafroll and Red Blotch Disease as Model Systems Monica L. Cooper and Malcolm B. Hobbs ............................................................................174 • Seasonal Ecology and Transmission Efficiency of Three-Cornered Alfalfa Hopper and Other Novel Insect Vectors of Grapevine Red Blotch Virus Kent M. Daane, Houston Wilson, Alexis Billings, and Kelsey Schall ..................................182 • Quantifying Vine Mealybug Spatio-Temporal Dynamics: Assessing Invasion Risk to Refine Management Strategies Matt Daugherty, Monica Cooper, and Aubrey May ..............................................................193 • Ecology of Grapevine Red Blotch Virus Marc Fuchs, Keith Perry, and Deborah Golino .....................................................................200 • I. Resistance to Grapevine Leafroll-Associated Virus 3 and the Grape Mealybug Marc Fuchs, Angela Douglas, Greg Loeb, and Deborah Golino ...........................................209 • II. Resistance to Grapevine Leafroll-Associated Virus 3 and the Grape Mealybug Marc Fuchs, Angela Douglas, Greg Loeb, and Deborah Golino ...........................................222 • Virus-Based Delivery of Interfering RNAs Targeting Grapevine Leafroll- Associated Virus(es) and Associated Mealybugs Yen-Wen Kuo and Bryce W. Falk .........................................................................................229 • Timing of Field Transmission of Grapevine Red Blotch Virus Robert R. Martin ....................................................................................................................235 • Identification of Grape Cultivars and Rootstocks with Resistance to Vine Mealybug Rachel Naegele ......................................................................................................................241 - iii - 2019 Research Progress Reports • Investigation of the Impact of Grapevine Red Blotch Virus on Grape Ripening and Metabolism Anita Oberholster ...................................................................................................................246 • Effects of Grapevine Red Blotch Disease on Flavor and Flavor Precursor Formation in the Grape and on Wine Quality Michael Qian and Alexander D. Levin ..................................................................................262 • Structure-Function Studies on Grapevine Red Blotch Virus to Elucidate Disease Etiology Christopher D. Rock and Sunitha Sukumaran .......................................................................272 • Understanding Symptomology and Physiological Effects of Red Blotch Disease in Vineyards in Oregon’s Willamette Valley Patty Skinkis and Robert R. Martin .......................................................................................292 • Improved Understanding of Virus Transmission of Grapevine Red Blotch Virus Vaughn Walton, Rachele Nieri, and Rick Hilton ..................................................................300 • Biology and Role of Treehoppers in Grapevine Red Blotch Disease Frank Zalom and Mysore R. Sudarshana ...............................................................................303 ABSTRACTS • Monitoring Vine Mealybug Resistance to Imidacloprid Mark Sisterson, Lindsey Burbank, Rachel Naegele, and Christopher Wallis .......................317