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.

Autor: Sumner-Kalkun JC; SASA, Edinburgh, United Kingdom., Sjölund MJ; SASA, Edinburgh, United Kingdom., Arnsdorf YM; SASA, Edinburgh, United Kingdom., Carnegie M; SASA, Edinburgh, United Kingdom., Highet F; SASA, Edinburgh, United Kingdom., Ouvrard D; Department of Life Sciences, Natural History Museum, London, United Kingdom.; Entomology and invasive plants Unit, Plant Health Laboratory, ANSES, Montferrier-sur-Lez Cedex, France., Greenslade AFC; Rothamsted Insect Survey, Rothamsted Research, Harpenden, Hertfordshire, United Kingdom., Bell JR; Rothamsted Insect Survey, Rothamsted Research, Harpenden, Hertfordshire, United Kingdom., Sigvald R; Department of Ecology, Swedish University of Agricultural Sciences, Uppsala, Sweden., Kenyon DM; SASA, Edinburgh, United Kingdom.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2020 Mar 26; Vol. 15 (3), pp. e0230741. Date of Electronic Publication: 2020 Mar 26 (Print Publication: 2020).
DOI: 10.1371/journal.pone.0230741
Abstrakt: 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.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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