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ARS Home » Pacific West Area » Wapato, Washington » Temperate Tree Fruit and Vegetable Research » Research » Publications at this Location » Publication #404695
Research Project: Developing New Potatoes with Improved Quality, Disease Resistance, and Nutritional Content

Location: Temperate Tree Fruit and Vegetable Research

Title: A high-throughput plate method for nucleic acid extraction from beet leafhopper (Hemiptera: Cicadellidae) and potato psyllid (Hemiptera: Triozidae) for pathogen detection

Author
item GORMAN, CHRISTOPHER - Washington State University
item CROWDER, DAVID - Washington State University
item Swisher Grimm, Kylie

Submitted to: Journal of Economic Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/24/2023
Publication Date: 8/16/2023
Citation: Gorman, C., Crowder, D.W., Swisher Grimm, K.D. 2023. A high-throughput plate method for nucleic acid extraction from beet leafhopper (Hemiptera: Cicadellidae) and potato psyllid (Hemiptera: Triozidae) for pathogen detection. Journal of Economic Entomology. 116(5):1876-1884. https://doi.org/10.1093/jee/toad153.
DOI: https://doi.org/10.1093/jee/toad153

Interpretive Summary: Pathogens that are transmitted to plants by insect pests can cause serious damage to crops each year if the pathogen or pest is not adequately controlled by a grower. In collaboration with Washington State University Scientists, a USDA-ARS scientist from Prosser, Washington, developed a high-throughput nucleic acid extraction method capable of generating high quality DNA for subsequent pathogen detection. Success of this method was demonstrated on the potato psyllid and beet leafhopper, which transmit the economically devastating 'Candidatus Liberibacter solanacearum' and 'Ca. Phytoplasma trifolii' pathogens, respectively. Optimization and validation of this new method increased processing efficiency 2.5-fold and nearly cut the cost for processing a single insect specimen in half. This new tool will enable researchers to generate pathogen prevalence data in a timely fashion for proper dissemination of results to the grower for application in their insect pest management programs.

Technical Abstract: Plant pathogens that are transmitted by insect vectors cause considerable damage to crops when pests or pathogens are not detected early in the season and populations are not controlled. Knowledge of pathogen prevalence in insect pest populations can aid growers in their insect pest management decisions but requires the timely dissemination of results. This process requires that specimen capture, identification, nucleic acid extraction, and molecular detection of pathogen(s) occur alongside a platform for sharing results. The potato psyllid (Bactericera cockerelli, Sulc; Hemiptera: Triozidae) and beet leafhopper (Circulifer tenellus, Baker; Hemiptera: Cicadellidae) transmit pathogens to potato and other vegetable or seed crops each season in the northwestern United States. While the potato psyllid has been tested for pathogen occurrence for the past decade, testing of the beet leafhopper is a new endeavor and substantially increases the specimen number that must be tested by our laboratories each season. To aid in the rapid processing of these insect specimens, we optimized and validated a new high-throughput 96-well plate nucleic acid extraction method for use in place of a standard 1.5ml single tube extraction method. Processing efficiency improved 2.5-fold and the cost associated with processing a single sample was nearly cut in half with this newly developed plate nucleic acid extraction method. Overall, this method has proven to be an excellent tool for the rapid testing of large numbers of small insect vectors to enable timely dissemination of data on pathogen prevalence to growers.