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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. and Sjolund, M. J. and Arnsdorf, Y. M. and Carnegie, M. and Highet, F. and Ouvrard, D. and Greenslade, A. F. C. and Bell, J. R. and Sigvald, Roland and Kenyon, D. M. (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. PLoS ONE. 15 , e0230741 , 1-18
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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.

Authors/Creators:Sumner-Kalkun, J. C. and Sjolund, M. J. and Arnsdorf, Y. M. and Carnegie, M. and Highet, F. and Ouvrard, D. and Greenslade, A. F. C. and Bell, J. R. and Sigvald, Roland and Kenyon, D. M.
Title: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
Year of publishing :2020
Volume:15
Article number:e0230741
Number of Pages:18
Publisher:PLoS
ISSN:1932-6203
Language:English
Publication Type:Journal article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 401 Agricultural, Forestry and Fisheries > Agricultural Science
URN:NBN:urn:nbn:se:slu:epsilon-p-106520
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-106520
Additional ID:
Type of IDID
DOI10.1371/journal.pone.0230741
Web of Science (WoS)000535934200051
ID Code:17229
Faculty:NJ - Fakulteten för naturresurser och jordbruksvetenskap
Department:(NL, NJ) > Dept. of Ecology
(S) > Dept. of Ecology
Deposited By: SLUpub Connector
Deposited On:10 Jul 2020 09:48
Metadata Last Modified:30 Aug 2020 17:55

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