利用DNA池技术提高基于InDel标记的种子纯度鉴定效率  

Improving Seed Purity Identification on InDel Pyrosequencing by DNA Pooling Technology

作　　者：兰青阔[1] 余景会[1] 赵新[1] 王永[1] 张桂华[2] 朱珠[1] 陈锐[1] 李欧静[1] 郭永泽[1] 程奕[1] 

机构地区：[1]天津市农业科学院中心实验室,天津300381 [2]天津科润农业科技股份有限公司黄瓜研究所,天津300192

出　　处：《种子》2012年第2期31-33,38,共4页Seed

基　　金：天津市应用基础及前沿技术研究计划(项目编号:10 JCYBJC09300);天津市科技支撑项目(项目编号:10 ZCGYNC00400)

摘　　要：为提高基于InDel-Pyrosequencing的黄瓜杂交种纯度检测通量,降低检测成本,本研究模拟10种DNA Pooling进行PCR、Pyrosequencing及等位基因频率分析。通过TTest分析不同Pooling间等位基因频率的差异性,确定3 Pooling为种子纯度检测最适Pooling数;建立3 Pooling-InDel-Pyrosequencing标准曲线,其R2达0.999 1;根据该标准曲线,检测黄瓜杂交品种"园中王"30粒杂交种种子纯度,结果为96.67%。本研究丰富了基于InDel-Pyrosequencing的黄瓜杂交种纯度检测技术体系。In order to reduce the seed purity identification cost,DNA pooling technology was combined with InDel-Pyrosequencing to improve the seed purity identification efficiency.The author simulated 10 DNA poolings for PCR,pyrosequencing.The result indicated that 3 pooling was suitable for seed purity identification.And 3 pooling-InDel-Pyrosequencing standard curve and its R2=0.999 1 were builded.Seed purity of Yuan Zhongwang were tested by 3 pooling-InDel-Pyrosequencing based on the standard curve,and the result was 96.67%.The study enriched the seed purity identification method based on InDel-Pyrosequencing.

关 键 词：DNA池 INDEL标记 焦磷酸测序技术 黄瓜 种子纯度鉴定 

分 类 号：S642.2[农业科学—蔬菜学]