丹参转录组的微卫星位点(SSR)特征及属内通用引物的开发

Development and Characterization of Microsatellite Markers for the East Asia Salvia Group using Transcriptome Sequencing

唇形科(Lamiaceae)鼠尾草属(Salvia)是一个具有丰富物种和高度多样性的类群,具有很高的科学研究与经济价值。该属可利用的通用SSR引物有限,因而从其代表物种丹参(Salvia miltiorrhiza)中开发新的通用性好的SSR引物十分必要。本研究利用丹参的15.99 Mb转录组数据,分析了含有Unigene的33438条序列,共获得2095个SSR候选位点,其包括了570个等位基因,每个位点的等位基因数范围在2~18个,平均每个位点7.92个。多态信息含量(PIC)范围在0.049至0.936之间,平均为0.707。根据设定条件选取了150个产物大小介于100~200 bp的引物,对属内20个物种进行通用性与多态性的检测,成功开发出通用性好,多态性丰富的72对引物。这些验证物种代表了鼠尾草属的3个不同亚属,可用于属内物种的遗传关系及其生态遗传学研究,具有较广泛的利用前景。

Salvia L.(Lamiaceae)is a rich and diverse taxonomic group that has important research and economic values.The limited availability of microsatellites for the genus Salvia necessitated the development of new markers from representative species(Salvia miltiorrhiza).Here,we examined 33438 Unigenes and 2095 simple sequence repeats(SSRs)containing candidate sequences were isolated from 15.99 Mb transcriptome profiling of S.miltiorrhiza.A total of 570 alleles were identified,with a number of alleles ranging from 2 to 18 and an average of 7.92 alleles per locus.The Polymorphism Information Content(PIC)ranged from 0.049 to 0.936 with a mean of 0.707.The study tested 150 loci primers that met primer design criteria and produced amplicons in the 100~200 bp range,and further examined their transferability and polymorphismin for 20 species.Seventy-two polymorphic EST-SSR markers were developed and could be amplified for all twenty Salvia species which belong to three subgenera:Sclarea Benth.,Salvia Benth.and Allagospadonopsis Briq.respectively.Our results high light the usefulness of mining pyrosequenced EST collections of gene-based SSRs to provide molecular markers that can examine genetic relationships and ecological genetics.