A diagnostic real-time PCR assay for the rapid identification of the tomato-potato psyllid, Bactericera cockerelli (Šulc, 1909) and development of a psyllid barcoding database

Sumner-Kalkun, J.C.; Sjölund, M.J.; Arnsdorf, Y.M.; Carnegie, M.; Highet, F.; Ouvrard, D.; Greenslade, A.F.C.; Bell, J.R.; Sigvald, R.; Kenyon, D.M.

Abstract

The accurate and rapid identification of insect pests is an important step in the prevention and control of outbreaks in areas that are otherwise pest free. The potato-tomato psyllid Bactericera cockerelli (Šulc, 1909) is the main vector of ‘Candidatus Liberibacter solanacearum’ on potato and tomato crops in North America and New Zealand; and is considered a threat for introduction in Europe and other pest-free regions. This study describes the design and validation of the first species-specific TaqMan probe-based real-time PCR assay, targeting the ITS2 gene region of B. cockerelli. The assay detected B. cockerelli genomic DNA from adults, immatures, and eggs, with 100% accuracy. This assay also detected DNA from cloned plasmids containing the ITS2 region of B. cockerelli with 100% accuracy. The assay showed 0% false positives when tested on genomic and cloned DNA from 73 other psyllid species collected from across Europe, New Zealand, Mexico and the USA. This included 8 other species in the Bactericera genus and the main vectors of ‘Candidatus Liberibacter solanacearum’ worldwide. The limit of detection for this assay at optimum conditions was 0.000001ng DNA (~200 copies) of ITS2 DNA which equates to around a 1:10000 dilution of DNA from one single adult specimen. This assay is the first real-time PCR based method for accurate, robust, sensitive and specific identification of B. cockerelli from all life stages. It can be used as a surveillance and monitoring tool to further study this important crop pest and to aid the prevention of outbreaks, or to prevent their spread after establishment in new areas.

Citation

Sumner-Kalkun, J., Sjölund, M., Arnsdorf, Y., Carnegie, M., Highet, F., Ouvrard, D., …Kenyon, D. (2020). A diagnostic real-time PCR assay for the rapid identification of the tomato-potato psyllid, Bactericera cockerelli (Šulc, 1909) and development of a psyllid barcoding database. PloS one, 15(3), Article e0230741. https://doi.org/10.1371/journal.pone.0230741

• Journal Article Type: Article
• Acceptance Date: Mar 6, 2020
• Publication Date: Mar 26, 2020
• Deposit Date: Feb 6, 2024
• Journal: PLOS ONE
• Print ISSN: 1932-6203
• Publisher: Public Library of Science
• Peer Reviewed: Peer Reviewed
• Volume: 15
• Issue: 3
• Article Number: e0230741
• DOI: https://doi.org/10.1371/journal.pone.0230741
• Publisher URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0230741