利用SSR技术鉴定玉米杂交种“家佳荣2号”的种子纯度  

Seed Purity Identification for Maize Hybrid 'Jiajiarong No.2' by SSR Marker Technique

作　　者：李阳[1] 范梦伟 季晓坤 贾相初 吴彬 赵自仙[1] 王德海[5] LI Yang;FAN Meng-wei;JI Xiao-kun;JIA Xiang-chu;WU Bin;ZHAO Zi-xian;WANG De-hai(College of Agriculture and Biotechnology,Yunnan Agricultural University,Yunnan Kumning 650201,China;Yunnan Shengyan Seed Co.Ltd.,Yunnan Yuxi 653100,China;Zhenxiong Seed Management Station,Yunnan Zhenxiong 657200,China;Kaiyuan Plant Pro-tection Station,Yunnan Kaiyuan 661699,China;Yunnan Seed Management Station,Yunnan Kumning 650031,China)

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]云南盛衍种业有限公司,云南玉溪653100 [3]镇雄县种子管理站,云南镇雄657200 [4]开远市植检植保站,云南开远661699 [5]云南省种子管理站,云南昆明650031

出　　处：《西南农业学报》2018年第7期1349-1354,共6页Southwest China Journal of Agricultural Sciences

基　　金：云南省技术创新人才培养项目(2011CI061);云南农业大学百名人才培养项目(2012PY03);云南农业大学博士启动基金项目(A2002276)

摘　　要：【目的】玉米杂交种子的纯度直接影响农作物的产量、品质和农户的收成,建立一套简单、快速、准确、可靠的纯度鉴定方法十分必要。【方法】本试验以‘家佳荣2号’玉米杂交种及其父母本为材料,采用幼叶CTAB法和碱煮法提取DNA两种方法,利用SSR标记技术从20对玉米纯度鉴定标准SSR引物中筛选应用于‘家佳荣2号’种子纯度鉴定的引物,同时人为掺入其他品种籽粒以验证特异引物的可靠性。此外,同步采用公认的田间小区种植鉴定法研究‘家佳荣2号’的种子纯度,比较室内和田间种植鉴定的纯度差异。【结果】采用幼叶CTAB法提取得到的DNA比碱煮法效果更好;筛选得到引物phi072k4和bnlg2291k4是‘家佳荣2号’种子纯度鉴定的特异性引物,掺杂验证结果可靠,‘家佳荣2号’室内SSR鉴定纯度为96.4%,田间种植鉴定的纯度为97.5%,2种方法结果高度一致。【结论】幼叶CTAB法提取DNA更适合玉米种子纯度的分子鉴定;SSR方法可用于玉米杂交种纯度的室内快速鉴定。【Objective】Maize hybrid seed purity affected yield,grain quality and farmers＇ income directly. It was necessary to establish a simple,rapid,accurate and reliable seed purity testing method for maize hybrids.【Method】Three materials（ Hybrid ‘Jiajiarong No. 2 ＇,male and female seeds）,two DNA extraction methods（ Leaves CTAB DNA extraction,Alkali Cooking DNA extraction） were used in this study. The primers were screened out to identify seed purity of the maize hybrid‘Jiajiarong No. 2＇from 20 SSR standard primers,and then another hybrid seeds were mixed deliberately into‘Jiajiarong No. 2＇to verify the reliability of specific primers. An authoritative seed purity identification method,field plot identification method,was applied to compare the purity identification result with laboratory testing.【Result】Leaves CTAB DNA extraction method was appropriate for maize SSR seed purity identification than Alkali Cooking DNA extraction,and there were two pair primers phi072 k4 and bnlg2291 k4 were specific primers for‘Jiajiarong No. 2＇,other hybrid seeds which were mixed into‘Jiajiarong No. 2＇could be identified successfully by this two specific primers. The purities of‘Jiajiarong No. 2＇were 96. 4 % and 97. 5 %respectively by SSR markers and field plot identification.【Conclusion】Leaves CTAB DNA extraction method is more appropriate for maize seed purity identification,SSR method can be used to identify fleetly of maize hybrid purity.

关 键 词：玉米 SSR标记 纯度 DNA提取 

分 类 号：S513[农业科学—作物学]