金线莲RAPD-SCAR标记的开发和种质遗传多样性评价

目的研究不同种质资源金线莲Anoectochilus roxburghii的遗传进化关系,开发高效、实用的分子标记方法。方法在20个不同产地金线莲种质资源的基础上开发RAPD标记,并将其换成特异SCAR标记。结果从100条RAPD随机引物中筛选出28条具有显著多态性的引物,扩增得到135条多态性位点。RAPD聚类分析显示,当20个金线莲种质资源平均遗传距离为0.748时,可以聚为6类,同一区域起源金线莲基本归为一类,表明不同区域金线莲种质资源存在着显著遗传差异。在此基础上,从多态性RAPD条带中挑选出5个特异位点转换为SCAR标记,并在不同金线莲种质资源中进行验证,结果显示5条SCAR标记具有显著的种质资源特异性和扩增模式。结论所筛选的特异RAPD-SCAR标记为加快金线莲优良品种选育进程奠定坚实基础。

Development of RAPD-SCAR marker for Anoectochilus roxburghii and their utilization to assess genetic diversity of germplasm

Objective To reveal the genetic relationship of germplasms of Anoectochilus roxburghii and develop an effective and valuable molecular marker. Methods In this study the polymorphic random amplified polymorphic DNA(RAPD) and specific SCAR markers were developed based on 20 different germplasms from various places. Results Twenty-eight 100 RAPD primers have significant polymorphism, generated 135 polymorphic bands among 20 germplasms. On the basis of RAPD results, 20 germplasms were clustered into six groups on genetic distance of 0.748. Conclusion Clustering analysis shows that there are the significant genetic differences among germplasms derived from different regions. A total of five specific bands from RAPD results are transferred into sequence characterized amplified region(SCAR) markers. Amplified results of SCAR markers among different germplasms show that SCAR markers are significant specific to different germplsms. This study has laid a solid foundation for accelerating the breeding of A. roxburghii.