| 刘秀丽1, 2, 庞博1, 张金文1*, 王蒂1*, 张俊莲1.双重PCR检测马铃薯晚疫病和环腐病方法的建立[J].植物保护,2015,41(2):114-119. |
| 双重PCR检测马铃薯晚疫病和环腐病方法的建立 |
| Development of duplex PCR assay for detection of Phytophthora infestans and Clavibacter michiganensis subsp.sepedonicum |
| 投稿时间：2014-02-23 修订日期：2014-06-11 |
| DOI： |
| 中文关键词: 马铃薯 双重PCR 马铃薯晚疫病菌 马铃薯环腐病菌 |
| 英文关键词:potato duplex PCR Phytophthora infestans Clavibacter michiganensis subsp. sepedonicum |
| 基金项目:甘肃省科技重大专项(1102NKDA025); 国家自然科学基金(31260343) |
|
| 摘要点击次数: 1175 |
| 全文下载次数: 690 |
| 中文摘要: |
| 通过克隆马铃薯环腐病菌和晚疫病菌转录间隔区(ITS)序列, 并对测序结果进行同源性比较, 选取差异位点分别设计了两对引物 P.IN1/P.IN2和C.IN1/C.IN2, 并检测了引物的特异性及方法的灵敏度。引物 P.IN1/P.IN2可扩增出1 条 363 bp 马铃薯晚疫病菌的特异性条带, 在 DNA 水平上其灵敏度达18 fg/μL; 引物C.IN1/C.IN2可扩增出1条218 bp 马铃薯环腐病菌的特异性条带, 在细菌数上检测灵敏度为104 cfu/mL。混合这两对引物构建双重PCR反应体系, 能从马铃薯环腐病菌和晚疫病菌的混合 DNA 及感染这两种菌的马铃薯植株中同时扩增到 363 bp和218 bp的特异片段。实现了同时对马铃薯晚疫病菌和环腐病菌的快速可靠检测。 |
| 英文摘要: |
| Based on cloning and analysis of the internal transcribed spacer (ITS) sequences of Phytophthora infestans and Clavibacter michiganensis subsp. sepedonicum, two specific pairs of primer, P.IN1/P.IN2 and C.IN1/C.IN2, were designed to establish a duplex PCR system, with the optimized PCR parameters to ensure the sensitivity and specificity. The system could simultaneously detect the two pathogens of potato. Using P.IN1/P.IN2, a single unique PCR band of 363 bp was amplified from P.infestans and the sensitivity was 18 fg/μL of DNA. Using C.IN1/C.IN2, a single unique PCR band of 218 bp was amplified from C.michiganensis subsp. sepedonicum and the sensitivity was 104 cfu/mL of bacteria. With the duplex PCR system, the 363 bp and 218 bp PCR bands from P.infestans and C.michiganensis subsp. sepedonicum could be specifically amplified. So a duplex PCR system has been developed for simultaneously and rapidly detecting P.infestans and C.michiganensis subsp. sepedonicum. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
