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

A - Papers appearing in refereed journals

Sumner-Kalkun, J. C., Sjolund, M. J., Arnsdorf, Y. M., Carnegie, M., Highet, F., Ouvrard, D., Greenslade, A. F. C., Bell, J. R., Sigvald, R. and Kenyon, D. M. 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), p. e0230741. https://doi.org/10.1371/journal.pone.0230741

AuthorsSumner-Kalkun, J. C., Sjolund, M. J., Arnsdorf, Y. M., Carnegie, M., Highet, F., Ouvrard, D., Greenslade, A. F. C., Bell, J. R., Sigvald, R. and Kenyon, D. M.
Abstract

The accurate and rapid identification of many 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 Central and Northern America and New Zealand. 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 successfully detected B. cockerelli genomic DNA from adults (100% accuracy, n=72); immatures (100% accuracy, n=26) and eggs (100% accuracy, n=25) . This assay also detected DNA from cloned plasmids containing the ITS2 region of B. cockerelli (100% accuracy, n=24). The assay showed 0% false positives when tested on genomic and cloned DNA from 73 other psyllid species collected from across Europe, New Zealand and Mexico. 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.

KeywordsBactericera cockerelli ; Biosecurity; Phytosanitary; Diagnostic; Pest; Vector; 15 ‘Candidatus Liberibacter solanacearum’; Real-time PCR
Year of Publication2020
JournalPLOS ONE
Journal citation15 (3), p. e0230741
Digital Object Identifier (DOI)https://doi.org/10.1371/journal.pone.0230741
Open accessPublished as green open access
FunderBiotechnology and Biological Sciences Research Council
Scottish Government
Funder project or codeThe Rothamsted Insect Survey - National Capability [2017-2022]
RRL/001/14
Publisher's version
Output statusPublished
Publication dates
Online26 Mar 2020
Publication process dates
Accepted06 Mar 2020
PublisherPublic Library of Science (PLOS)
ISSN1932-6203

Permalink - https://repository.rothamsted.ac.uk/item/9755x/a-diagnostic-real-time-pcr-assay-for-the-rapid-identification-of-the-tomato-potato-psyllid-bactericera-cockerelli-ulc-1909-and-development-of-a-psyllid-barcoding-database

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