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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Sustainable Perennial Crops Laboratory » Research » Publications at this Location » Publication #394382

Research Project: Development of Pathogen- and Plant-Based Genetic Tools and Disease Mitigation Methods for Tropical Perennial Crops

Location: Sustainable Perennial Crops Laboratory

Title: A new hundred-genomes pipeline enables rapid duplex LAMP detection of two bacterial spot pathogens

item BERAN, PAVEL - University Of South Bohemia
item STEHLIKOVA, DAGMAR - University Of South Bohemia
item Cohen, Stephen
item ROST, MICHAEL - University Of South Bohemia
item BERANOVA, KRISTINA - University Of South Bohemia
item CURN, VLADISLAV - University Of South Bohemia

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/22/2022
Publication Date: 11/26/2022
Citation: Beran, P., Stehlikova, D., Cohen, S.P., Rost, M., Beranova, K., Curn, V. 2022. A new hundred-genomes pipeline enables rapid duplex LAMP detection of two bacterial spot pathogens. Plant Disease.

Interpretive Summary: Correctly identifying plant diseases in a timely manner is crucial for managing crops. Here, we demonstrate our newly developed method for correctly and rapidly identifying disease-causing bacteria. Using a software-based approach, we were able to analyze hundreds of publicly available bacterial DNA sequences. This let us quickly design a field test that can be used to monitor for two bacteria that cause disease in tomato and pepper plants. Our results give a new disease-monitoring strategy to growers and extension agents.

Technical Abstract: Xanthomonas euvesicatoria and X. vesicatoria are two economically important causal agents of bacterial spot (BS) of tomato and pepper. Management of BS in the field requires rapid and accurate detection. To enable molecular detection of these two pathogens in the field, we developed a new pipeline for producing duplex LAMP assays. We analyzed 109 publicly available bacterial genomic sequences to design an assay suitable for field detection of the two pathogens. We confirmed both specificity and sensitivity of the assay by testing the complex of bacterial strains pathogenic to tomato and pepper. We also confirmed that the assay was suitable for use on plants infected by the pathogens. The assay detection limit was 1 pg of genomic DNA with an assay duration of 40 minutes on a mobile detection platform. The high specificity, sensitivity, and speed of the assay will allow it to be deployed, contributing to successful management of the BS.