一种基于HRM技术的安全及高通量水稻光敏不育基因分型体系的创建与应用  

Development and Application of an HRM-based,Safe and High-throughput Genotyping System for Photoperiod Sensitive Genic Male Sterility Gene in Rice

作　　者：司浩杰 汪庆 刘洋洋 黄建中[1] 舒庆尧[1] 谭瑗瑗 

机构地区：[1]浙江大学农业与生物技术学院作物研究所/国家水稻生物重点实验室,浙江杭州310058 [2]无锡哈勃生物种业技术研究院,江苏无锡214100 [3]浙江大学新农村发展研究院,浙江杭州310058

出　　处：《核农学报》2017年第11期2081-2086,共6页Journal of Nuclear Agricultural Sciences

基　　金：浙江省农业(粮食)新品种选育重大科技专项(2016C02050-2);杭州市重大科技创新项目(2015012A09)

摘　　要：为建立适用于大规模应用功能性分子标记辅助选择(MAS)选育光周期敏感核雄性不育(PGMS)水稻的安全、高通量和廉价的光敏基因分型体系,本研究通过将基于竞争性扩增差异熔解扩增子(CADMA)的高分辨熔解曲线(HRM)分析与水稻基因组DNA快速提取方法相结合,创建了一种安全、廉价和高通量的PGMS基因分型体系。利用该基因分型体系,从约8 000株花培苗中检测出了2 027株PGMS水稻,并对117个DH_1光敏不育单株的DH_2材料进行跟踪检测,检出了异交单株。在该体系中,96个水稻材料的取样、DNA提取和PGMS基因分型可在2.5 h左右完成。从样本DNA提取到基因分型均不用有毒化学物质,所有操作均在基于96孔PCR板的操作平台进行,闭管操作、无交叉污染。本研究建立的PGMS基因分型体系必将极大提高水稻光敏不育系选育的效率。A safe, high throughput and low-cost genotyping system for PGMS gene was developed to facilitate the breeding of PGMS rice using molecular assisted selection （ MAS）. This paper established a genotyping system for PGMS gene by integrating a rapid DNA extraction into CADMA-HRM analysis. 2 027 PGMS plants were identified from about 8 000 plants generated from anther culture with this system. Besides the PGMS and homozygous wild-type, heterozygous plants were detected among DH2 progenies from 117 DH1 plants. DGMS genotyping of 96 samples could be completed within 2.5 h including sample collecting and DNA extraction. All the operations from DNA extraction to genotyping were performed in a 96 - well plate, no toxic chemicals and cross contamination. The established system for PGMS genotyping would remarkably enhance the efficiency of PGMS breeding.

关 键 词：水稻 PGMS HRM CADMA 

分 类 号：S511[农业科学—作物学]