利用多重PCR技术快速检测4种水稻病原细菌

A multiplex PCR method for rapid detection of four rice pathogenic bacteria

对水稻中多种病原细菌的检测,使用常规方法往往耗时耗力,而多重PCR可以更加高效地进行多种细菌的检测。根据水稻细菌性谷枯病菌gyrB基因,水稻细菌性叶鞘褐腐病菌PfsI/R quorum sensing位点以及水稻细菌性条斑病菌和水稻白叶枯病菌含铁细胞接受因子基因设计引物,建立4种水稻病菌的多重PCR检测方法,对方法进行特异性和灵敏度测试,并对采自不同地区的水稻样本进行检测。结果显示,多重PCR方法能同步地快速检测出水稻细菌性谷枯病菌、水稻细菌性叶鞘褐腐病菌、水稻细菌性条斑病菌或水稻白叶枯病菌,检测灵敏度达到103 cfu/mL的菌液浓度,利用该方法对我国不同地区的58份水稻种子进行检测,其中17个样本检测出水稻细菌性条斑病菌或水稻白叶枯病菌,未检测到水稻细菌性谷枯病菌和水稻细菌性叶鞘褐腐病菌。

Multiplex PCR is an efficient and simple method to detect a variety of pathogenic bacteria in rice at the same time.A multiplex PCR detection method was established for four rice pathogenic bacteria,and the primers were designed based on the gyrB gene of Burkholderia glumae,the PfsI/R quorum sensing locus of Pseudomonas fuscovaginae and the iron-containing cell-receiving factor genes of Xanthomonas oryzae pv.oryzae and X.oryzae pv.oryzicola.The specificity and sensitivity of the multiplex PCR method were validated,and rice samples from different regions were tested using this method.The results showed that the multiplex PCR method can fast and accurately detect four rice pathogenic bacteria.The detectability of this method reached 103 cfu/mL in the bacterial solution.Among 58 rice samples from different regions,neither B.glumae nor P.fuscovaginae was detected,while X.oryzae pv.oryzae or X.oryzae pv.oryzicola were positive in 17 samples.