利用多重PCR诊断水稻白叶枯病和细菌性条斑病的复合发生

Dignosis of complex infection of Xanthomanas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola by Multi-plex PCR

为准确检测水稻白叶枯病菌、细菌性条斑病菌及这两种病菌的复合发生,利用软件DNAStar分析比较这两种菌的部分核酸序列,设计了检测这两种病菌的特异性引物。引物Xoo F-Xoo R能特异性扩增出水稻白叶枯病菌中一条大小162 bp的条带;引物Xooc F1-Xooc R1和Xooc F2-Xooc R2能够分别特异性扩增出水稻细菌性条斑病菌中690 bp和945 bp的条带。通过优化PCR反应条件,成功建立了多重PCR技术,可以对不同国家的水稻白叶枯病菌和细菌性条斑病菌进行准确检测,对由这两种病菌引起的复合侵染实现了准确诊断。

In order to detect Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola and co-infection of these two pathogens,partial DNA sequences of X. oryzae pv. oryzae and X. oryzae pv. oryzicola were analyzed and the specific primers were designed by software DNAStar. A 162 bp fragment could be amplified from X. oryzae pv. oryzae with the primer pair Xoo F-Xoo R while 690 bp and 945 bp fragment could be amplified from X. oryzae pv. oryzicola with the primer pair Xooc F1-Xooc R1 and Xooc F2-Xooc R2 respectively. After optimization of the PCR condition,multi-plex PCR was successfully developed,with which could detect X. oryzae pv. oryzae and X. oryzae pv. oryzicola from different countries precisely. The result demonstrated that multi-plex PCR could detect and identify the co-infection of these two pathogens.