Development of a Genomics-Based LAMP (Loop-Mediated Isothermal Amplification) Assay for Detection of Pseudomonas fuscovaginae from Rice.

Autor: Ash GJ; Graham Centre for Agricultural Innovation (an alliance between Charles Sturt University and NSW DPI), School of Agricultural and Wine Sciences, Charles Sturt University, Wagga Wagga 2678, NSW, Australia., Lang JM; Colorado State University, Department of Bioagricultural Sciences & Pest Management, Fort Collins 80523-1177., Triplett LR; Colorado State University, Department of Bioagricultural Sciences & Pest Management, Fort Collins 80523-1177., Stodart BJ; Graham Centre for Agricultural Innovation., Verdier V; Institut de Recherche pour le Développement, UMR Résistance des Plantes aux Bioagresseurs, IRD-CIRAD-UM2, 34394 Montpellier Cedex 5, France., Cruz CV; International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines., Rott P; CIRAD, UMR BGPI, F-34398 Montpellier, France., Leach JE; Colorado State University, Department of Bioagricultural Science & Pest Management.
Jazyk: angličtina
Zdroj: Plant disease [Plant Dis] 2014 Jul; Vol. 98 (7), pp. 909-915.
DOI: 10.1094/PDIS-09-13-0957-RE
Abstrakt: The vast amount of data available through next-generation sequencing technology is facilitating the design of diagnostic marker systems. This study reports the use of draft genome sequences from the bacterial plant pathogen Pseudomonas fuscovaginae, the cause of sheath brown rot of rice, to describe the genetic diversity within a worldwide collection of strains representing the species. Based on a comparative analysis with the draft sequences, primers for a loop-mediated isothermal amplification (LAMP) assay were developed to identify P. fuscovaginae. The assay reported here reliably differentiated strains of P. fuscovaginae isolated from rice from a range of other bacteria that are commonly isolated from rice and other plants using a primer combination designated Pf8. The LAMP assay identified P. fuscovaginae purified DNA, live or heat-killed cells from pure cultures, and detected the bacterium in extracts or exudates from infected host plant material. The P. fuscovaginae LAMP assay is a suitable diagnostic tool for the glasshouse and laboratory and could be further developed for in-field surveys.
Databáze: MEDLINE