乌头及其同属近缘种的SSR指纹图谱研究

SSR Fingerprinting for Aconitum carmichaeli Debx. and Its Relatives

乌头(Aconitum carmichaeli Debx.)为中国重要的药用植物,种质资源丰富,同属近缘种繁多。该研究采用SSR分子标记技术对采自17个种群的51份乌头样本和13个同属近缘种的65份样本进行扩增和聚丙烯酰胺凝胶电泳检测,分析了乌头及其同属近缘种的遗传多样性、遗传分化和系统发育关系,筛选出稳定性好、多态性高的SSR引物构建乌头及其近缘种的指纹图谱。结果表明:(1)11对乌头微卫星引物均表现出高的多态性,共检测出109个复等位基因,平均每个位点9.91。(2)乌头在物种水平上遗传多样性丰富(A=3.090 9,I=0.889 7,h=0.540 2),种群间的遗传分化显著(Gst=0.277 4);乌头属14种植物表现出了较高的遗传多样性(A=9.909 1,I=1.526 2,h=0.690 5),物种间遗传分化显著(Fst=0.437),基因流微弱(Nm=0.451 8)。(3)聚类分析表明,同一种群的乌头样本首先聚在一起各自形成小支,17个小支聚为一支;13个乌头近缘种材料中,相同物种的样本分别聚为一支;乌头属14个物种聚为5个大支,与形态分类结果一致。(4)利用基因型丰富、多态性高的6对SSR引物(Tchin03、Tchin04Tchin20、Tchin26、Tchin29、Tchin32)可有效区分14种乌头属植物,并以此建立了乌头种质资源和近缘种的DNA指纹图谱。该研究为乌头的种质资源鉴定及混伪品鉴定提供了重要的技术支撑。

Aconitum carmichaeli Debx.is an important medicinal herb in China,it has abundance germplasm and relatives in the same genus.In this study,we employed SSR markers to analyses51individuals of A.carmichaeli.sampled from17populations and its65relatives from13species in genus Aconitum L..Genetic diversity,genetic differentiation and phylogenetic relationship of the individuals and species were revealed.As the results,(1)11SSR primer pairs used in this study are stable and polymorphic,109alleles were screened out totally,with the average of9.91per pair.(2)The genetic diversity of A.carmichaeli is abundance(A=3.0909,I=0.8897,h=0.5402),the genetic differentiation among populations is obvious(Gst=0.2774).High polymorphism(A=9.9091,I=1.5262,h=0.6905),pronounced genetic differentiation(Fst=0.437)and limited gene flow among species(Nm=0.4518)were detected in the14species in genus Aconitum.(3)Cluster analysis revealed that most of the A.carmichaeli individuals sampled from the same population clustered together formed17small clades,and then formed a clade;as for those relatives,samples of same species clustered together and form different clades individually;14species of genus Aconitum formed5big clades,which is consistent with the result of traditional taxonomy.(4)6primer pairs(Tchin03,Tchin04Tchin20,Tchin26,Tchin29and Tchin32),which performed higher polymorphic and more abundance genotype were chosen to construct the fingerprinting for A.carmichaeli germplasms and its relatives.The study provides reliable molecular basis for classification and identification for A.carmichaeli germplasm and its relatives in the same genus.