摘要
该研究对高山杜鹃(Rhododendron L.)的总DNA提取方法进行了改良,然后综合利用高山杜鹃EST数据库和该实验室的马缨杜鹃高通量测序数据,引用已发表文献的SSR引物,从154对SSR引物中筛选出了26对多态性高、重复性好、条带清晰的SSR引物。再从中随机选择10对SSR引物进行荧光标记,对69份不同高山杜鹃的种质进行遗传多样性分析。结果表明,平均有效等位基因数6.959 2个;位点多态性信息含量(PIC)、观测杂合度(HO)、期望杂合度(HE)和Neis基因多样性(H)分别为0.795 2、0.543 5、0.826 5和0.820 2;杂交种间的非加权配对算术平均(UPGMA)聚类结果与其谱系分析结果基本一致,可实现对一些未知来源的育成品种资源进行祖先亲本类型的推测。
In this study,the total DNA extraction method was modified based on the conventional CTAB method for Rhododendron,and then many microsatellite sequences were chosen from the published Rhododendron EST database and unpublished genome database of R.delavayi in our lab to design SSR primer pairs.Moreover,adding many SSR primer pairs referred to the references,154 pairs of SSR primers were used to be screened.Finally,26 pairs of polymorphic primers were screened out,and all of PCR amplicons were specific with clearer band and better repeatability.Furthermore,10 pairs of polymorphic SSR primer fluorescently-labeled were used to genetic diversity analysis on 69 Rhododendrongermplasms.The results as following：mean effective allele number（Ne）,polymophism information content（PIC）,observed heterozygosity（HO）,expective heterozygosity（HE）,and Nei＇s gene diversity（H）for 10 SSR locus are 6.959 2,0.795 2,0.543 5,0.826 5and 0.820 2,respectively.Based on the consistency between the UPGMA clustering results and their pedigree analysis of Rhododendronhybrids,a speculation of ancestor parent types will be made for Rhododendron hybrids with unknown or unclear pedigree.
出处
《西北植物学报》
CAS
CSCD
北大核心
2016年第12期2421-2432,共12页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(31460217
31560225)
云南省应用基础研究计划(2016FB058)
云南省花卉育种重点实验室开放基金(FKL-201501)
云南省科技计划(2015BB013)
关键词
高山杜鹃
DNA提取
SSR引物筛选
系谱
亲缘关系
alpine Rhododendron
DNA extraction
SSR primer screening
pedigree
genetic relationship
