黄淮产区地黄种质遗传多样性分析及SSR指纹图谱构建

Genetic Diversity Analysis and SSR Fingerprint Construction of Germplasms of Rehmannia glutinosa in Huang-Huai Region

为明确我国黄淮产区地黄种质间遗传多样性,解析种质间的亲缘关系,为该区地黄品种选育和种质鉴定提供依据,利用毛细管电泳技术和108对SSR引物对地黄种质进行分析,以获取种质间遗传相似系数、多态性引物、种质分辨率,利用软件TASSEL 3.0和Fig Tree(V 1.4.3)进行聚类分析和作图,选取多态性好、稳定性强的引物构建地黄种质的SSR指纹图谱。结果表明,共获得多态性引物50对,其片段长度在107~365 bp,种质间遗传相似系数在0.552~0.984,平均为0.729,野生种质的平均遗传相似系数明显低于地方品种和当前主栽品种。聚类结果显示,17个地黄种质分为3个亚群。选取并利用多态性强和分辨率高、扩增稳定的15对引物构建了地黄种质的SSR指纹图谱,实现了对每个地黄种质的快速鉴定。综上,SSR分析是实现地黄鉴真、聚类、DNA指纹图谱构建的一种有效方法。

The main objectives of this study were to clarify the genetic diversity among germplasms of Rehmannia glutinosa in Huang-Huai production area of China,and understand the genetic relationship among germplasms or accessions,so as to provide a basis for breeding and germplasm identification of Rehmannia glutinosa.108 pairs of SSR primers were used to amplify germplasms and the results were detected by capillary electrophoresis to obtain the genetic similarity coefficients,and count the polymorphic primer information and resolution.SSR fingerprints of those germplasm were constructed by selecting primers with good polymorphism and stability.The softwares TASSEL 3.0 and Fig Tree(V 1.4.3)were used for cluster analysis and mapping.The results showed that 50 pairs of polymorphic primers were obtained and fragment length ranged from 107 bp to 365 bp.The variation range of genetic similarity coefficient among germplasms was 0.552—0.984,and the average value was 0.729.The average genetic similarity coefficient of wild germplasms was significantly lower than that of landraces and cultivated varieties.The clustering results showed that 17 germplasms could be divided into three subgroups.By using 15 pairs of primers with stable polymorphism and high resolution,the SSR fingerprint of Rehmannia glutinosa Germplasms was constructed,and the rapid identification of germplasms from the DNA level was realized.SSR analysis is an effective method to distinguish,cluster Rehmannia glutinosa and construct DNA fingerprint.