EST-SSR构建益母草栽培种指纹图谱及遗传多样性

DNA Fingerprinting Establishment and Genetic Diversity Analysis of Leonurus japonicus Cultivars with EST-SSR

为了对益母草品种进行分子鉴别及分析不同品种间的亲缘关系,试验采用EST-SSR(Expressed Sequence Tag-Simple Sequence Repeats)分子标记技术构建了生产上常用的10个益母草栽培品种的数字指纹图谱及遗传多样性分析。试验共筛选出13对益母草EST-SSR分子标记,检测等位位点61个,其中含多态性位点52个,多态性比率为85.2%。EST-SSR引物扩增2~11个等位位点,平均每对引物扩增4.69个基因型,扩增条带100~600 bp,多态信息含量(PIC)0.551,基因流(Nm)1.319,平均杂合率0.647。从13对引物中,筛选出5对核心引物,即LJ17、LJ21、LJ29、LJ47和LJ62,以引物组合方式构建指纹图谱可区分10个品种;聚类分析表明在遗传距离0.66处10个品种被分为3类,聚类结果与品种的来源具有较高的一致性。

To identify the cultivars of Leonurus japonicus and analyze the genetic relationship among different cultivars at a molecular level,the authors constructed the digital fingerprint for 10 common Leonurus japonicus cultivars using expressed sequence tag-simple sequence repeats(EST-SSR)molecular markers.The 24 pairs of primers amplified a total of 61 alleles including 52 polymorphic alleles with a polymorphism rate of 85.2%.Each pairs of primers can amplify 2-11 alleles,with an average of 4.69 alleles per locus.The length of amplified products was 100-600 bp,with the mean value of polymorphism information content(PIC)being 0.654,Nm(gene flow)being 1.319,and the mean value of Ho(heterozygosity)being 0.647.Among the 13 primers,5 primers can be used as a preferred choice for fingerprinting,namely LJ17,LJ 21,LJ 29,LJ 47 and LJ 62,in which 5 primers can be employed in combination to identify the 10 cultivars.Cluster analysis show that the 10 cultivars can be divided into three groups at a genetic distance of 0.66;the genetic relationship as indicated by the cluster completely reflected the place of origin of the cultivars.