AIMS: The aim of this study is to develop a multiplex PCR (mPCR) assay for rapid, sensitive and simultaneous detection of 6 important rice pathogens: Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, Pseudomonas fuscovaginae, Burkholderia glumae, B. gladioli and Acidovorax avenae subsp. avenae. METHODS AND RESULTS: Specific primers were designed through a bioinformatics pipeline. Sensitivity of detection was established using both traditional PCR and quantitative real-time PCR on isolated DNA and on bacterial cells both in vitro and in simulated diseased seeds and parameters were optimized for an mPCR assay. A total of 150 bacterial strains was tested for specificity. The mPCR assay accurately predicted the presence of pathogens amongst 44 symptomatic and asymptomatic rice seed, sheath and leaf samples. CONCLUSIONS: This study confirmed that this mPCR assay is a rapid, reliable and simple tool for the simultaneous detection of 6 important rice bacterial pathogens. This article is protected by copyright. All rights reserved.
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