猕猴桃EPIC标记开发及其在猕猴桃属植物系统发育分析中的应用

Development of EPIC Markers in Actinidia and Their Application for Phylogenetic Analysis of Genus Actinidia

采用生物信息学方法通过比较ESTs数据和基因组数据开发了猕猴桃的双亲遗传单拷贝的EPIC标记以用于猕猴桃属植物系统发育研究。通过比较中华猕猴桃（Actinidiachinensis）和美味猕猴桃（4．deliciosa）的ESTs数据和葡萄（Iqtisvinifera）基因组数据，以一致性85％为标准共检测到129个EPIC标记，并从中设计96对引物。其中21对EPIC引物在中华猕猴桃和美味猕猴桃两个商业化栽培品种中呈现稳定扩增，核酸多态性（Pj）为0．003～0．069，变异位点为0．7％-14．8％，简约性信息位点为0～9．4％。用其中的EPIC．1标记重建中华猕猴桃、美味猕猴桃、毛花猕猴桃（Aeriantha）、绵毛猕猴桃（Afulvicom口var．1anata）和金花猕猴桃（4．chrysantha）的系统发育关系，显示其跨种间扩增的实用性并获得分辨率较高的系统发育树。本研究中开发的EPIC标记可以作为通用引物用于猕猴桃属植物系统发育分析。

In the present study, a set ofbiparental and single-copy EPIC markers were developed by blasting ESTs database of Actinidia chinensis and A. deliciosa and genome database of lOtis vinifera. A total of 129 EPIC markers were identified with an average identity /〉 85% in the flanking exon regions. A total of 96 EPIC primer pairs were designed and tested, of which, 21 successfully amplified the target regions in A. chinensis and A. deliciosa. The nucleotide diversity （Pi）, variable sites, parsimony informative sites varied from 0.003 to 0.069, 0.7% to 14.8%, and 0 to 9.4%, respectively. Phylogeny of five Actinidia species using EPIC-1 marker was reconstructed, showing the primers developed here could be successfully amplified in A. eriantha, A. fulvicoma var. lanata and A. chrysantha. The results indicatedthat EPIC markers have good cross-species transferability and can be applied to phylogenetic analysis in Actinidia as universal primers.