百日草属EST-SSR信息分析及其雄性不育两用系鉴别引物筛选  

EST-SSR Resource Analysis of Zinnia and Primer Screening for the Identification of Its Male Sterile Two-type Lines

作　　者：胡妙 秦美姣 李娟[1] 叶要妹[1] 

机构地区：[1]华中农业大学园艺林学学院/湖北省林业信息工程技术研究中心/园艺植物生物学教育部重点实验室,湖北武汉430070

出　　处：《江西农业大学学报》2017年第5期976-982,共7页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(30771518);中央高校基本科研业务费项目(2662011PY101);国家级大学生创新创业训练计划项目(105042016033)

摘　　要：百日草雄性不育在F1代杂交育种中应用广泛,建立百日草雄性育性EST-SSR分子标记,为百日草分子标记辅助育种奠定科学基础。本研究通过分析NCBI数据库中百日草属EST序列20 667条,进行EST-SSR信息的分析。采用20μL红酶PCR反应体系,以百日草雄性不育两用系MJ16AB的DNA为模板,筛选引物。结果获得622条重复基元不少于5次且重复序列长度≥12 bp的SSR,其中二、三核苷酸出现次数最多的是AT/TA、CTA/GAT,分别占总数的9.5%、10.6%。随机合成的136对引物中,引物28、30、33、69和Ze M63在百日草可育和不育两池间扩增出清晰且稳定的差异性条带。本试验验证了基于百日草属EST数据开发SSR分子标记的可行性,筛选出的5对多态性引物可用于建立百日草雄性育性分子标记。Male sterility of Zinnia elegans is widely used in F1 generation. In order to establish the EST-SSR molecular markers of Zinnia elegans male fertility,provide the scientific foundation for the molecular marker assisted breeding of Zinnia elegans,the EST sequences of Zinnia from NCBI were analyzed,20 667 unigenes were obtained to develop new EST-derived SSR markers.Then with 20 μL red enzyme PCR reaction,taking the male sterile two-type lines MJ16 AB of Zinnia elegans as templates,primers were screened 622 SSRs,whose repeated motifs were not less than 5 times and repeated length greater than 12 bp,were obtained.Among them,the dominant types of the dinucleotide and trinucleotide repeats were AT/TA and CTA/GAT,accounting for 9.5% and 10.6% repectively.5 among 136 pairs of primers could produce stable and expected polymorphism PCR products between fertile pool and sterile pool of Zinnia elegans,they were primers 28,30,33,69 and Ze M63 respectively.The results indicated that it is a feasible way to develop EST-SSR markers from Zinnia elegans EST,5 polymorphic primers could be used for the establishment of molecular markers of Zinnia elegans male fertility.

关 键 词：百日草 雄性不育两用系 EST-SSR PCR反应体系 引物筛选 

分 类 号：S681.9[农业科学—观赏园艺]