Abstract: Early and rapid identification of F1 hybrid progeny and acquisition of true hybrids are of great significance to the breeding and genetic research of Trifolium repens superior strains. Six phenotypic traits of 2 Trifolium repens parent materials and 55 F1 hybrids were mapped by clustering heat map,and 3 pairs of specific SSR primers were selected to identify the authenticity of F1 hybrids. The results showed that the 57 materials in the cluster heat map could be divided into 2 classes according to the phenotypic characters of their parents. A total of 281 distinct bands and 137 polymorphic bands were obtained by amplification with 19 pairs of selected primers. The polymorphic sites accounted for 48.41%. The PIC of each SSR site ranged from 18.49% to 43.58%,with an average value of 34.73%. A total of 53 true-hybrids were identified by three pairs of specific primers,and the true-hybrid ratio was 96.36%,which was basically consistent with the results of UPGMA cluster analysis. The 19 pairs of SSR molecular marker primers have high polymorphism,which can be directly used for genetic diversity analysis of Trifolium repens and hybrid identification and other related studies. The identified F1 generation true hybrids of Trifolium repens can lay a foundation for subsequent breeding work.

Key words: Trifolium repens, Identification of Hybrid, SSR molecular marker, Genetic diversity, Phenotypic analysis

摘要： 早期快速鉴定白三叶(Trifolium repens L.)F1杂交子代，获取真杂种对白三叶优良品系的培育和遗传学研究具有重要的意义。本文针对两个白三叶亲本及55个杂交F1的6个表型性状进行分析，绘制聚类热图，再选用3对特异性SSR引物鉴定杂交F1代的真实性。结果表明，在聚类热图中57个株系可以根据亲本的表型性状划分为父本型和母本型。筛选出的19对SSR引物共扩增共得到清晰可辨条带281条，多态性条带137条，多态位点占比为48.41%，每个SSR位点的PIC在18.49%~43.58%之间，平均值为34.73%。其中3对特异性引物在55个杂交后代中共鉴定出53个真杂种，真杂种比率为96.36%，与UPGMA聚类分析结果基本一致。19对SSR分子标记引物具有较高的多态性，可直接用于白三叶遗传多样性分析、杂种鉴定等相关研究，鉴定到的白三叶F1代真杂种为后续育种工作奠定重要基础。

关键词: 白三叶, 杂种鉴定, SSR分子标记, 遗传多样性, 表型分析