摘要
以中华猕猴桃(Actinidia chinensis Planch)矮型种质‘赣猕5号’为母本,普通型中华猕猴桃‘奉雄2号’为父本构建F1群体,采用优越而高效的基于表达序列标签(Expressed Sequence Tags,EST)的简单序列重复(Simple Sequence Repeats,SSR)标记技术,结合群体分离分析法(Bulked Segregant Analysis,BSA)在荧光DNA测序仪(ABI3130)上进行了‘赣猕5号’矮型性状连锁的EST-SSR分子标记研究。结果表明,通过本实验室开发并设计的85对EST-SSR荧光引物的扩增筛选,其中引物EST-Ad042在亲本及其矮型与普通型DNA池之间扩增出一条285bp的多态性片段,经对其F1代分离群体部分矮型与普通型单株的随机验证,该片段稳定出现,在矮型植株中表现为有带,在普通型植株中表现为无带。遗传连锁分析表明,该标记与矮型基因之间的连锁距离为8.8cM。EST-SSR荧光引物能够有效地筛选到中华猕猴桃的特异标记,可以作为鉴别矮型与普通型植株的标记引物。
High effective simple sequence repeats（SSR）marker based on expressed sequence tags （EST）and the method of bulked segregant analysis（BSA）on the automated fluorescent-labeled system （ABI3130）were employed to screen the molecular marker linked to dwarf gene in kiwifruit （Actinidia spp.） from the F1 separating progenies obtained from the cross between dwarf cultivar ‘Ganmi 5’（♀） and normal cultivar‘Fengxiong 2’（♂）of Actinidia chinensis Planch. Eighty-five pairs of fluorescent primers were designed and screened in the present study. The results showed that a polymorphic band of 285 bp amplified by primer EST-Ad042 was occured both in the dwarf parent and in the dwarf cultivars of separating DNA pools but not in normal cultivar. Furthermore,the polymorphic band had been approved by stochastic amplification in the F1 separating progenies including dwarf and normal genotypes, respectively. The genetic distance between the EST-SSR marker and dwarf gene could be 8.8 cM by genetic linkage analysis. The specific molecular marker of kiwifruit could be effectively mined by fluorescent EST-SSR primers,and the fluorescent primer EST-Ad042 could be utilized toidentify its dwarf plant. The screening of molecular markers linked to dwarf gene in Actinidia chinensis Planch had been provided a basis for the studies of dwarf gene mapping and cloning in kiwifruit breeding program.
出处
《园艺学报》
CAS
CSCD
北大核心
2010年第4期553-558,共6页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(30660113
30860167)
作者简介
E-mail：xiaobiaoxu@hotmail．com
