黄瓜棒孢叶斑病菌PCR检测方法的建立

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

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**

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**

Invstitute of Vegetables and Flowers，Chinese Academy of Agricultural Sciences，Beijing 100081，China

Online:2014-03-25 Published:2014-03-25

摘要/Abstract

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

中图分类号:

S 642.2

陈璐, 石延霞, 谢学文, 柴阿丽, 李宝聚. 黄瓜棒孢叶斑病菌PCR检测方法的建立[J]. 园艺学报, 2014, 41(3): 585-592.

CHEN Lu, SHI Yan-Xia, XIE Xue-Wen, CHAI A-Li, LI Bao-Ju. PCR Assay for Detection of Corynespora cassiicola，the Causal Agent of Corynespora Leaf Spot of Cucumber[J]. ACTA HORTICULTURAE SINICA, 2014, 41(3): 585-592.

[1]	罗文彬，李华伟，汤浩，邱思鑫，纪荣昌，许泳清，刘中华，邱永祥. 马铃薯5种病毒多重PCR检测技术的建立及应用[J]. 园艺学报, 2015, 42(2): 280-288.
[2]	梁云, 袁素霞, 冯慧颖, 徐雷锋, 袁迎迎, 刘春, 明军. 百合肌动蛋白基因lilyActin 的克隆与表达分析[J]. 园艺学报, 2013, 40(7): 1318-1326.
[3]	周晓云, 游春平. 红掌根腐病病原鉴定及其PCR 检测方法[J]. 园艺学报, 2013, 40(5): 989-.
[4]	刘翠兰, 燕丽萍, 毛秀红, 孙超, 夏阳, 梁慧敏. 基因枪介导红叶石楠遗传转化因素分析[J]. 园艺学报, 2013, 40(11): 2280-2286.
[5]	赵英;牛建新. 梨叶片中苹果褪绿叶斑病毒的引物原位标记检测[J]. 园艺学报, 2009, 36(1): 15-20.