Skip to main content
Research article - Peer-reviewed, 2020

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

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.; 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 (Sulc, 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 (similar to 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.

Published in

PLoS ONE
2020, volume: 15, number: 3, article number: e0230741
Publisher: PUBLIC LIBRARY SCIENCE

Authors' information

Sumner-Kalkun, J. C.
SASA
Sjolund, M. J.
SASA
Arnsdorf, Y. M.
SASA
Carnegie, M.
SASA
Highet, F.
SASA
Ouvrard, D.
Nat Hist Museum
Ouvrard, D.
ANSES
Greenslade, A. F. C.
Rothamsted Res
Bell, J. R.
Rothamsted Res
Swedish University of Agricultural Sciences, Department of Ecology
Kenyon, D. M.
SASA

Associated SLU-program

SLU Network Plant Protection

UKÄ Subject classification

Agricultural Science

Publication Identifiers

DOI: https://doi.org/10.1371/journal.pone.0230741

URI (permanent link to this page)

https://res.slu.se/id/publ/106520