青枯菌为应对逆境胁迫,可进入活的但非可培养状态(viable but non-culturable,VBNC)。本文利用叠氮溴化丙锭(PMA)与PCR技术相结合,建立了一种快速有效区分青枯菌死活细胞的分子检测方法。基于hrcS基因序列,设计了一对青枯菌种特异性检测引物hrcSf/hrcSr;利用PMA对青枯菌Po82菌株的细胞悬浮液样品进行预处理,随后进行常规PCR扩增。结果表明,当样品中PMA质量浓度为3μg/mL、曝光时间大于5min时,PMA可有效抑制死亡菌体细胞中的DNA扩增;且对可培养和VBNC状态细胞中的DNA扩增没有影响;本试验建立的PMA-PCR方法能有效对包括VBNC状态在内的青枯菌活菌进行检测,避免了假阳性与假阴性结果的产生。
【英文摘要】
Ralstonia solanacearum can enter into the VBNC state(viable but non-culturable)in response to adverse environmental conditions.A novel method to differentiate viable/dead cells of R.solanacearum was established by using a DNA dye of propidium monoazide(PMA)in combination with the polymerase chain reaction(PMA-PCR).On the basis of hrcS gene sequence,a species-specific primer set hrcSf/hrcSr was designed for detection of R.solanacearum.Samples were pre-treated with PMA which bound DNA from dead cells so that ...
(1.College of Horticulture and Landscape,Tianjin Agricultural University,Tianjin 300384,China;2.Institute of Plant Protection,Chinese Academy of Agricultural Sciences,State Key Laboratory forBiology of Plant Diseases and Insect Pests,Beijing 100193,China)
| 
推荐 
点击下列相关研究机构和相关文献作者，可以直接查到这些机构和作者被《中国知识资源总库》收录的其它文献，使您全面了解该机构和该作者的研究动态和历史。
