为了对软枣猕猴桃栽培品种进行分子鉴别及分析不同品种间的亲缘关系,采用对叶片直接进行PCR扩增的方法,利用EST-SSR分子标记技术构建了常见的42个软枣猕猴桃品种的数字指纹图谱和模式指纹图谱数据库(cluster project),并进行了遗传多样性分析.结果表明:利用21对软枣猕猴桃核心引物扩增所有品种的基因组,共检测到132个等位位点,包括122个多态性位点,多态性比率为92.4%.每对引物扩增出的等位位点数为3~8个,平均每对引物扩增6.28个基因型,扩增条带大小100~300bp,多态信息含量(polymorphism information content,CPIC)为0.721,基因流为1.454,平均杂合率为0.691.其中,7对引物,即ACAR2,ACAR13,ACAR53,ACAR75,ACAR92,ACAR112和ACAR120可作为指纹图谱构建的首选引物,利用其中的ACAR2,ACAR13,ACAR53和ACAR75以引物组合方式构建指纹图谱可区分42个品种.聚类分析表明在遗传距离0.64处42个品种被分为一类,在遗传距离0.81处分为4类.聚类结果与品种的特性及来源具有较高的一致性.
In order to identify the cultivars of Actinidia arguta and analyze the genetic relationship among different cultivars at a molecular level,we constructed the digital fingerprint and model fingerprint(ClusterProject)for 42 common A.arguta cultivars using EST-SSR(expressed sequence tag-simple sequence repeats)molecular markers.The 21 pairs of primers amplified a total of 132 alleles including 122 polymorphic alleles with a polymorphism rate of 92.4%.Each pairs of primers can amplify 3~8alleles,with an average of 6.28 alleles per locus.The length of amplified products was100~300bp,with the mean value of CPIC(polymorphism information content)being 0.721,Nm(gene flow)being 1.454,and the mean value of Ho(heterozygosity)being 0.691.Among the 21 primers,7primers can be used as a preferred choice for fingerprinting,namely ACAR2,ACAR13,ACAR53,ACAR75,ACAR92,ACAR112 and ACAR120,in which 4primers(ACAR2,ACAR13,ACAR53 and ACAR75)can be employed in combination to identify the 42 cultivars.Cluster analysis showed that the42 cultivars can be divided into one group at a genetic distance of 0.64,and four groups at a distance of0.81.The genetic relationship as indicated by the cluster completely reflected the characteristics and the source of the cultivars.