Genomics‐informed design of loop‐mediated isothermal amplification for detection of phytopathogenic Xanthomonas arboricola pv. pruni at the intraspecific level. (2nd July 2012)

• Record Type: Journal Article
• Authors: Bühlmann, A.; Pothier, J. F.; Tomlinson, J. A.; Frey, J. E.; Boonham, N.; Smits, T. H. M.; Duffy, B.
• Is Part Of: Plant pathology. Volume 62:Number 2(2013:Apr.)
• Page Start: 475
• Page End: 484
• Publication Date: 2012-07-02
• Subjects: Agricultural pests -- Periodicals; Plant diseases -- Periodicals; 632
• DOI: 10.1111/j.1365-3059.2012.02654.x
• Languages: English
• ISSNs: 0032-0862

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The objective of this study was to develop a rapid, sensitive detection assay for the quarantine pathogen <italic>Xanthomonas arboricola</italic> pv. <italic>pruni</italic>, causal agent of stone fruit bacterial spot, an economically important disease of <italic>Prunus</italic> spp. Unique targets were identified from <italic>X. arboricola</italic> pv. <italic>pruni</italic> genomes using a comparative genomics pipeline of other <italic>Xanthomonas</italic> species, subspecies and pathovars, and used to identify specific diagnostic markers. Loop‐mediated isothermal amplification (LAMP) was then applied to these markers to provide rapid, sensitive and specific detection. The method developed showed unrivalled specificity with the 79 tested strains and, in contrast to previously established techniques, distinguished between phylogenetically close subspecies such as <italic>X. arboricola</italic> pv. <italic>corylina</italic>. The sensitivity of this test is comparable to that of a previously reported TaqMan^™ assay at 10³ CFU mL⁻¹, while the unrivalled speed of LAMP technology enables a positive result to be obtained in &lt;15 min. The developed assay can be used with real‐time fluorescent detectors for quantitative results as well as with DNA‐staining dyes to function as a simplified strategy for on‐site pathogen detection.</p> </abstract>