Annals of Applied Biology (2013) 163, 378-393

M. Slawiak, R. van Doorn, M. Szemes, A.G.C.L. Speksnijder, M. Waleron, J.M. van der Wolf, E. Lojkowska and C.D. Schoen (2013)
Multiplex detection and identification of bacterial pathogens causing potato blackleg and soft rot in Europe, using padlock probes
Annals of Applied Biology 163 (3), 378-393
Abstract: The objective of this study was to develop a multiplex detection and identification protocol for bacterial soft rot coliforms, namely Pectobacterium wasabiae (Pw), Pectobacterium atrosepticum (Pba) and Dickeya spp., responsible for potato blackleg and tuber soft rot. The procedures were derived from the phylogenetic relationships of these and other Enterobacteriaceae based on recA sequences. The group of Pw strains was highly homogeneous and could be distinguished from the other species. A ligation-based method for detection of Pw was developed. Five padlock probes (PLPs) were designed, targeting recA sequences to identify the Pw, Pba or Dickeya spp., whereas a sixth probe recognised recA sequences of all soft rot coliforms including Pectobacterium carotovorum subsp. carotovorum (Pcc). Two PLP-based applications were developed: one using real-time PCR and one using universal microarrays. Assay sensitivity and specificity were demonstrated using 71 strains of Pw, Pcc, Pba and Dickeya spp. Both multiplex methods can be potentially used for seed testing and in ecological studies, but further validation is required.
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Link to article at publishers website
Database assignments for author(s): Ewa Lojkowska, Jan M. van der Wolf, Malgorzata Waleron

Research topic(s) for pests/diseases/weeds:
surveys/sampling/distribution
identification/taxonomy

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Pectobacterium carotovorum	Potato (Solanum tuberosum)
	Pectobacterium atrosepticum	Potato (Solanum tuberosum)
	Pectobacterium wasabiae	Potato (Solanum tuberosum)