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园艺学报 ›› 2014, Vol. 41 ›› Issue (3): 585-592.

• 新方法 • 上一篇    下一篇

黄瓜棒孢叶斑病菌PCR检测方法的建立

陈 璐*,石延霞*,谢学文,柴阿丽,李宝聚**   

  1. 中国农业科学院蔬菜花卉研究所,北京 100081
  • 出版日期:2014-03-25 发布日期:2014-03-25
  • 基金资助:

    国家现代农业产业技术体系建设专项资金项目(CARS-25);农业部园艺作物生物学与种质创制重点实验室项目

PCR Assay for Detection of Corynespora cassiicola,the Causal Agent of Corynespora Leaf Spot of Cucumber

CHEN Lu*,SHI Yan-xia*,XIE Xue-wen,CHAI A-li,and LI Bao-ju**   

  1. Invstitute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China
  • Online:2014-03-25 Published:2014-03-25

摘要: 根据黄瓜棒孢叶斑病菌多主棒孢(Corynespora cassiicola)与棒孢属下女贞棒孢(Corynespora
lieustri)、威尔士棒孢(Corynespora cambrensis)和其他常见病原真菌Actin 基因序列差异,设计多主棒孢
的特异性引物Caa5F/Caa5R,建立了黄瓜棒孢叶斑病菌的PCR 检测方法,可对引起黄瓜棒孢叶斑病的病
原菌扩增出160 bp 的特异性条带,检测灵敏性为4 pg · μL-1 DNA,并可从接种后发病的黄瓜叶片总DNA
中检测到特异条带。该引物的PCR 检测方法灵敏性较高,可直接检测植株总DNA,无需病原菌的分离培
养,适用于对黄瓜棒孢叶斑病特异快速的检测。

关键词: 黄瓜棒孢叶斑病菌, 检测方法, Actin 基因, PCR 检测

Abstract: A rapid PCR detection assay of Corynespora cassiicola,the causal agent of Corynespora
leaf spot of cucumber,was developed based on the Actin gene of the pathogen. Primer pair Caa5F/Caa5R
were designed on the different bases of Actin gene sequences between C. cassiicola and other common
plant fungal pathogens,including Corynespora lieustri and Corynespora cambrensis,and only C.
cassiicola performed a 160 bp amplication. C. cassiicola could be detected in infected cucumber leaves
without isolation or cultivation by the PCR assay. The assay with a high sensitivity of 4 pg · μL-1 DNA per
reaction could be a useful tool of rapid detection of Corynespora leaf spot of cucumber.

Key words: Corynespora cassiicola, Actin gene, PCR

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