滇皖产区铁皮石斛居群SSR分子身份证的构建

摘要

目的:建立滇皖产区铁皮石斛SSR指纹图谱,并构建不同居群的SSR分子身份证.方法:使用生物信息学方法分析铁皮石斛(Dendrobium officinale)全基因组SSR(Simple sequence repeat)序列,获得SSR位点,使用Primer Premier 50软件设计引物并进行PCR扩增,选择高多态性引物进行遗传多样性分析并建立SSR指纹图谱代码,使用二维码生成器表述铁皮石斛居群的分子身份证.结果:共获得91 653个SSR位点,平均分布距离为1544 kb.据此开发了75对SSR引物,筛选出32对多态性丰富、带型清晰的引物,对11个滇、皖产区的铁皮石斛居群进行遗传多样性分析,共得到117个多态信息位点,平均每个SSR引物多态信息含量(PIC)为062,变化范围为022~080;铁皮石斛不同居群进行亲缘关系聚类表明11个铁皮石斛居群可分为三支,与种质来源分布相一致.筛选出4对多态性高、带型清晰易于统计的引物组合,建立不同居群铁皮石斛的SSR指纹图谱库和不同居群的分子身份证.结论:SSR分子身份证可为今后铁皮石斛的品种鉴别提供准确、可靠的方法和溯源信息平台.%Objective:To establish a molecular ID(mID)for Dendrobium officinale rigin from Yunnan and AnhuiMethods:Genome of Dofficinale was analyzed and SSRs (Simple Sequence Repeats) were isolatedPrimers were designed by Primer Premier 50 software,high polymorphism primers were isolated and genetic diversity analysis was performed,and thenSSR fingerprinting database and mIDs of 11 populations of Dofficinale were constructedResults:A total of 91 653 SSRs were isolated,the average distribution density was 1544 kb75 pairs of SSR primers were designed and screened,32 high polymorphismprimer pairs were selected to generate polymorphic fingerprintsThe results showed that the 32 pairs of primers could generate 117 polymorphic alleles bandsThe average polymorphism information content (PIC)as 062 with a range of 022-08011 populations of Dofficinale could be clustered to 3 branches,which accordance with the geographical distribution of its germplasmFingerprinting database of 11 populations of Dofficinale were established with 4 pairs of primers,and in consequence the molecular ID were established for the 11 populationsConclusion:SSR marker is suitable for construction of DNA fingerprinting database of Dofficinale and the database could provide reference for population identification of Dofficinale.