基于SSR标记构建中国芍药品种资源分子身份证

Molecular identification of herbaceous peony cultivars based on SSR markers

【目的】利用SSR标记区分中国芍药品种,为芍药资源的保护与合理利用奠定基础。【方法】对山东菏泽和河南洛阳2个主要芍药栽培地的品种进行调查,明确中国芍药品种资源数量,在此基础上,提取各品种叶片DNA,利用21对SSR荧光引物进行PCR扩增,用四重荧光毛细管电泳技术检测扩增片段长度。利用Microsatellite和Toolkit GenALEx 6.41软件,分析数据中的等位基因数、多态性信息含量(PIC)和遗传多样性参数。采用数字与大小写字母结合的方法,对不同长度的扩增片段进行赋值,构建中国芍药品种分子身份证。【结果】山东菏泽和河南洛阳目前共有芍药品种268个,用21对引物在这些品种中共扩增条带613条,多态信息含量0.508～0.846,平均为0.707;有效等位基因数(Ne)为4.324±0.315,观测杂合度(Ho)为0.593±0.047,期望杂合度(He)为0.741±0.020。采用21对SSR引物可将所有268个芍药品种区分开。【结论】采用21对引物通过SSR标记建立了山东菏泽和河南洛阳268个芍药品种的分子身份证,有助于芍药品种鉴别、新品种培育及防止资源流失。

[Objective] This study used SSR markers to distinguish Chinese herbaceous peony cultivars to lay foundation for rational use and protection of the herbaceous peony resources. [Method] Herbaceous peony cultivars were investigated in Heze and Luoyang,two main herbaceous peony cultivated areas. Based on quadruple fluorescence capillary electrophoresis,21 pairs of fluorescence primers were used to amplify all samples. Microsatellite and Toolkit GenALEx 6.41 were used to analyze allele number, polymorphism information content （PIC） and genetic diversity parameters. The molecular ID was then created with num bets,case letters and lowercase letters. [Result] A total of 268 varieties were analyzed in central China,the amount of amplified bands was 613 and the polymorphism information content was 0. 508 0. 846 with the average of 0. 707. The Ne was 4. 324±0. 315,the Ho was 0. 593±0. 047 and the He was 0. 741±0. 020. Those 21 pairs of primers can distinguish all the cultivars. [Conclusion] The specific molecular identity of Chinese peony cultivars was constructed based on 21 pairs of primers,which was of great significance for identification of peony varieties, prevention of resource loss and cultivation of new varieties.