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林业科学 ›› 2018, Vol. 54 ›› Issue (7): 51-61.doi: 10.11707/j.1001-7488.20180706

• 论文与研究报告 • 上一篇    下一篇

97个山杏无性系的遗传多样性及SSR指纹图谱

金玲, 刘明国, 董胜君, 吴月亮, 张欣   

  1. 沈阳农业大学林学院 沈阳 110161
  • 收稿日期:2017-07-26 修回日期:2017-12-18 出版日期:2018-07-25 发布日期:2018-08-11
  • 基金资助:
    国家林业公益性行业科研专项(201004034);辽宁特聘教授基金项目(2012)。

Genetic Diversity and Fingerprints of 97 Armeniaca sibirica Clones Based on SSR Markers

Jin Ling, Liu Mingguo, Dong Shengjun, Wu Yueliang, Zhang Xin   

  1. College of Forestry, Shenyang Agricultural University Shenyang 110161
  • Received:2017-07-26 Revised:2017-12-18 Online:2018-07-25 Published:2018-08-11

摘要: [目的]山杏是北方半干旱地区重要的生态经济型树种,种质资源异常丰富,形态鉴定与分类难度大。利用SSR分子标记研究山杏优选无性系的遗传多样性并构建指纹图谱,以期为山杏种质鉴定提供科学依据。[方法]根据山杏简化基因组测序结果,合成600对SSR引物,利用4个山杏无性系进行引物筛选,选出155对扩增条带清晰的引物,对97个山杏优选无性系进行PCR扩增。应用引物组合法构建指纹图谱,采用非加权组平均法(UPGMA)进行聚类分析。[结果]利用155对SSR引物对97个山杏无性系进行扩增,共扩增出933个等位基因,每个位点的等位基因数为3~11个,平均为6.019个;各位点的多态性信息含量(PIC)为0.476~0.885,平均值为0.681,具有较高的多态性。50个山杏无性系在59个位点上具有特异等位基因,89个无性系在131个位点上具有特异基因型。采用5对引物(L56、X47H、L79H、P40H和X47)的组合可区分全部97个无性系,构建了指纹图谱。对97个山杏无性系进行亲缘关系分析,无性系间的遗传相似系数变化范围为0.669~0.943,平均值为0.757;基于遗传相似系数进行聚类分析,将97个无性系分为5大类,第1大类和第2大类下又分为3个亚类,分类结果与无性系的来源区域有明显的相关性。[结论]从600对SSR引物中筛选出155对,其扩增位点多态性较高、重复性较好。发现89个山杏无性系具有特异基因型,其中50个既具有特异基因型也具有特异等位基因。应用引物组合法构建了基于5对SSR引物扩增位点的山杏无性系指纹图谱。供试山杏无性系遗传差异较小,亲缘关系较近。研究结果可为山杏种质鉴别提供科学依据,也为山杏的育种工作奠定基础。

关键词: 山杏, 无性系, SSR, 遗传多样性, 指纹图谱

Abstract: [Objective] Armeniaca sibirica is an important ecological and economic tree species. Its germplasm resources are very abundant. Identification and classification through morphological characteristics are difficult. In this paper, genetic diversity of superior clones of A. sibirica was analyzed and the fingerprints were constructed using SSR markers in order to provide a scientific basis for germplasm identification.[Method] Based on the results of simplified genome sequencing of A. sibirica, 600 pairs of SSR primers were synthesized, and screened for four A. sibirica clones. A total of 155 pairs of primers with clear bands were selected and used for PCR amplification for the 97 A. sibirica clones. The fingerprints were constructed by primer combination method and cluster analysis was carried out using UPGMA.[Result] Using 155 pairs of SSR primers, a total of 933 alleles were generated in the 97 A. sibirica clones. The number of alleles at each locus ranged from 3 to 11, with an average of 6.019. PIC of these loci ranged from 0.476 to 0.885, with an average of 0.681, indicating that the 155 loci were highly polymorphic. A total of 50 A. sibirica clones had specific alleles at 59 loci, and 89 clones had specific genotypes at 131 loci. All the 97 clones could be distinguished by the combination of primer L56, X47H, L79H, P40H and X47, and the fingerprints were constructed with the combinations. Genetic relationships of the 97 A. sibirica clones were analyzed, and the genetic similarity coefficients between the clones ranged from 0.669 to 0.943, with a mean of 0.757. Clustering analysis based on genetic similarity coefficients showed that the 97 clones could be divided into five categories, and the first category and the second category could be divided into three sub-categories respectively, which was consistent to a considerable extent with the clone origins.[Conclusion] In this study, 155 pairs of SSR primers were selected from 600 pairs, and the amplified loci had high polymorphism and good repeatability. It was found that 89 clones had specific genotypes, of which 50 had both specific genotypes and specific alleles. The fingerprints were constructed by the combination of 5 pairs of primers. The genetic differences among the clones were not significant, and the relationships among them were close. The results provide a scientific basis for germplasm identification of A. sibirica and lay a foundation for breeding of the species.

Key words: Armeniaca sibirica, clone, SSR, genetic diversity, fingerprint

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