水稻白叶枯病菌和水稻细菌性条斑病菌的实时荧光PCR快速检测鉴定

成功建立了水稻白叶枯菌与水稻细菌性条斑病菌快速检测鉴定的实时荧光PCR方法。根据含铁细胞接受子基因设计两菌的通用引物PSRGF/PSRGR（扩增一个152bpDNA片段）和特异性探针（Baiprobe和Tiaoprobe），并对13种细菌和1种植原体进行实时荧光PCR。结果表明，两个特异性探针能分别特异性检测到目标病原菌产生荧光信号而其它参考菌不产生荧光信号。检测的绝对灵敏度是30.6fg/μL质粒DNA和103CFU/mL的菌悬浮液，相当于1个细菌细胞的基因，比常规PCR电泳检测高约100倍，相对灵敏度为105CFU/mL。整个检测过程只需2h，完全闭管，降低了污染的机会，无需PCR后处理。 用这两个特异性探针分别对自然感染白叶枯菌和条斑菌的叶片DNA提取液和种子浸泡液进行实时荧光PCR，结果均可特异性检测到目标菌的存在并完全可将两种病原细菌区分开来,且只需03g叶片和10g种子。

Detection and Identification of Xanthomonas oryzae pv.oryzae and Xanthomonas oryzae pv.oryzicola by Real-time Fluorescent PCR

A novel and sensitive real time PCR was developed to detection \%Xanthomonas ory zae\% pv.\%oryzae\% and \%Xanthomonas oryzae\% pv.\%oryzicola\%, which cause the bacteria leaf blight (BLB) and leaf streak respectively, Universal and specific TaqMan probes, which were designed based on the sequence of Putative siderophor e receptor gene cds were used to detect 13 bacteria and one phytoplasmas, only in \%X.oryzae \%pv. \%oryzae\% and \%X.oryzae \%pv. \%oryzicola\%, fluorescen t signal can be collected with their specific probes respectively. The level of detection of the probe was 30.6fg plasmid, roughly equaling to one cell and 100 times sensit ive than PCR gel electrophoresis detection.\% X.oryzae \%pv. \%oryzae\% and \% X.oryzae \%pv \%oryzicola\% were detected from seed washes and DNA extracted fro m the seed washes of naturally infected seeds and in fected leaves as small as 10g naturally infected seeds or 0.3g leaf. This method is little time consumption (only 2h) and without contamination from PCR product .