中国农业科学2009，Vol.42Issue(12)：4403-4410,8.DOI:10.3864/j.issn.0578-1752.2009.12.036
实时荧光定量PCR鉴定小麦矮腥黑穗菌技术研究
Detection of Tilletia controversa Kuhn by Real Time Quantitative PCR
摘要
Abstract
[Objective] To detection of Tilletia controversa Kühn (TCK) sensitively and accurately,real-time PCR systems were developed.[Method]The species-specific primer pair CQUTCK_4/CQUTCK_5 and probe CQUP_1 were designed based on a selected specific fragment (Ⅰ322 bp) specific for TCK,and the SYBR Green Ⅰ and TaqMan quantitative PCR detection systems were established with optimized reaction conditions.[Result]The detection limit of the two systems were 0.1fg,equal to 2.31×10~4 copies,which was 10~2-10~3 fold higher than conventional PCR.By the constructed detection systems,the TCK and Tilletia caries (DC)Tul (TCT) could be distinguished.The teliospore and mycelium of TCK in the infected wheat plant tissue also could be identified accurately and rapidly.[Conclusion]The earlier diagnosis approaches of wheat durwf bunt pathogen were set up using the twe real-time PCR systems.关键词
TCK/TCT/实时荧光定量PCRKey words
TCK/TCT/Real-time Fluorescent Quantitative PCR分类
农业科技引用本文复制引用
年四季,袁青,殷幼平,蔡俊,王中康..实时荧光定量PCR鉴定小麦矮腥黑穗菌技术研究[J].中国农业科学,2009,42(12):4403-4410,8.基金项目
国家科技部"863"项目(2006AA10Z434)、农业部重点攻关项目(2006-37) （2006AA10Z434）
