文献类型：期刊文献

中文题名：油菜茎基溃疡病菌的实时荧光PCR检测

英文题名：Detection of Leptosphaeria maculans by real-time PCR

作者：周国梁[1];尚琳琳[2];林泓[1];印丽萍[1];徐殿胜[2];易建平[1]

机构：[1]上海出入境检验检疫局,上海200135;[2]华东理工大学生物工程学院,上海200237

年份：2011

卷号：41

期号：1

起止页码：10

中文期刊名：植物病理学报

外文期刊名：Acta Phytopathologica Sinica

收录：CSTPCD;;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

基金：上海出入境检验检疫局科研项目(HK069-2010);上海市科委项目(09DZ0580201)

语种：中文

中文关键词：油菜茎基溃疡病菌;实时荧光PCR

外文关键词：Leptosphaeria maculans; real-time PCR

摘要：根据油菜茎基溃疡病菌Leptosphaeria maculans与其近似种ITS序列的差异,设计了检测L.maculans的引物Lmb3/R2和探针Probe-M,建立了L.maculans的实时荧光PCR检测方法。试验结果表明,来自加拿大、澳大利亚和乌克兰等国的22株L.maculans菌株都能得到阳性扩增,而供试的30株L.biglobosa菌株和6株其他菌株以及空白对照没有荧光信号的增加。该检测方法的灵敏度达到4 pg菌丝DNA,整个检测过程控制在4 h内,其快速、特异和灵敏的特点可以满足进境油菜籽样品的快速初检以及病菌分离物的快速鉴定。
Real-time PCR was used for detection of Leptosphaeria maculans, the causative agent of oilseed phoma stem canker. Specific primers Lmb3/R2 and a TaqMan-MGB probe （Probe-M） specific to L. macu- lans were designed from the differentiation of ITS sequences between L. maculans and related species in the GenBank database. The results showed that all 22 isolates of L. maculans from oilseed samples from Austra- lia, Ukraine and Canada could get positive amplification with the limit of 4 pg DNA, and negative results for 30 isolates of L. biglobosa and the other 6 isolates of related species from oilseed samples. Real-time PCR could be especially useful for mass screening of oilseed sample and diagnosis of suspected isolate.