期刊
JOURNAL OF APPLIED MICROBIOLOGY
卷 120, 期 5, 页码 1357-1367出版社
WILEY
DOI: 10.1111/jam.13094
关键词
comparative genomics; detection; multiplex PCR; quantitative real-time PCR; rice bacterial pathogens
资金
- Special Fund for Agro-Scientific Research in the Public Interest [201303015]
- National Natural Science Foundation of China [31200003]
- 12th Five Years Key Programs for Science and Technology Development of China [2012BAK11B02, 2012BAK11B06]
- Zhejiang Provincial Nature Science Foundation of China [LY12C14007]
AimsThe aim of this study was to develop a multiplex PCR (mPCR) assay for rapid, sensitive and simultaneous detection of six important rice pathogens: Xanthomonas oryzae pv. oryzae, X.oryzae pv. oryzicola, Pseudomonas fuscovaginae, Burkholderia glumae, Burkholderia gladioli and Acidovorax avenae subsp.avenae. Methods and ResultsSpecific 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 invitro and in simulated diseased seeds and the parameters were optimized for an mPCR assay. A total of 150 bacterial strains were tested for specificity. The mPCR assay accurately predicted the presence of pathogens among 44 symptomatic and asymptomatic rice seed, sheath and leaf samples. ConclusionsThis study confirmed that this mPCR assay is a rapid, reliable and simple tool for the simultaneous detection of six important rice bacterial pathogens. Significance and Impact of the StudyThis study is the first report of a method allowing simultaneous detection of six major rice pathogens. The ability to use crude extracts from plants without bacterial isolation or DNA extraction enhances the value of this mPCR technology for rapid detection and aetiological/epidemiological studies.
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