实时荧光定量PCR技术在球毛壳ND35定殖检测中的应用  

Application of Quantitative Real-Time PCR in Colonization Detection of Chaetomium globosum ND35

作　　者：刘璇[1] 戴杨[1] 高克祥[1] 米士伟[1] 吴海燕[1] 何邦令[1] 

机构地区：[1]山东农业大学植物保护学院,山东泰安271018

出　　处：《山东农业科学》2012年第6期17-19,共3页Shandong Agricultural Sciences

基　　金：国家自然科学基金项目(30872024;30571498);公益性行业(农业)科研专项(201003004);山东省科技发展计划项目(2010GNC10911)

摘　　要：对球毛壳(Chaetomium globosum)ND35菌株的RAPD扩增特异性DNA片段进行分析,利用Prim-er 5.0软件设计了一对引物,即ND35-1:5'-GCTCGCAGCTCTGGTGCGGT-3',ND35-2:5'-GCCGATTGC-CCATGTCCACCTC-3',以球毛壳ND35菌株基因组DNA为模板,利用此引物建立并优化了SYBR GreenⅠ实时荧光定量PCR反应的最佳体系,并据此对杨树和黄瓜两种植物中ND35菌株的定殖情况进行了检测。结果表明,该引物可以用于球毛壳ND35的实时荧光定量PCR检测。Based on the analysis of RAPD specific DNA fragment of Chaetomium globosum ND35, one pair of primers （ ND35 - 1：5＇ - GCTCGCAGCTCTGGTGCGGT - 3＇, ND35 - 2 ： 5＇ - GCCGATYGCCCATGTC- CACCTC - 3＇） was designed by software Primer 5.0. Taking the genomic DNA of C. globosum ND35 as tem- plate, a system of quantitative real - time PCR with SYBR Green I was developed and optimized, and then it was used to detect the colonization of strain ND35 in cucumber and poplar. The results showed that this pair of specific primers could be used for the determination of C. globosum ND35.

关 键 词：球毛壳ND35 实时荧光定量PCR 定殖 检测 

分 类 号：Q503[生物学—生物化学]