基于SSR的刺槐无性系遗传多样性分析和指纹图谱构建

doi:10.11707/j.1001-7488.20171009

摘要/Abstract

摘要： [目的]对山东省刺槐无性系进行遗传多样性分析和指纹图谱构建，为刺槐新品种选育、遗传改良和品种鉴定提供可靠的依据，为中国其他省市刺槐遗传资源保存、评价与利用提供参考。[方法]采用荧光SSR引物进行PCR扩增，产物经毛细管电泳仪进行检测；利用所得结果分析49个无性系遗传多样性并构建其指纹图谱；采用非加权组平均法（UPGMA）进行基于亲缘关系的无性系聚类分析。[结果]8对SSR引物共检测到51个等位基因，每个位点的等位基因为2~15个，平均每对引物扩增出6.375个多态性等位基因。不同位点的PIC值变化范围很大，在0.092~0.879之间，平均PIC值为0.509 8。使用组间平均数联结法进行UPGMA聚类分析，结果表明49份无性系没有严格按照不同区域聚类到一起，无性系分组与现有栽培区没有明显的规律。来自临沂的‘鲁刺8’和‘鲁刺13’、青岛的‘鲁刺40’和‘鲁刺42’以及日照的‘鲁刺10’和‘鲁刺86’亲缘关系最近。本研究所使用的9对引物中，其中2对Rply109和rops16可以区分开45份无性系，鉴定率达到91.84%。检测到22个刺槐无性系在1个或者2个位点得到3个等位基因，表明这些无性系可能是发生自然变异的多倍体。[结论]山东省刺槐具有较高的遗传多样性。无性系分组与现有栽培区没有明显的规律。引物Rply109和rops16对49份无性系的分子鉴定率为91.84%，可作为刺槐指纹图谱构建和分子鉴定的高效分子标记。利用SSR标记构建的指纹图谱可为刺槐遗传资源管理、品种鉴定和知识产权保护奠定坚实的基础，同时为刺槐的引种和遗传育种亲本选择提供科学的理论依据。

关键词: 刺槐, SSR, 分子标记, 遗传多样性, 指纹图谱, 聚类分析

Abstract: [Objective]To analyze genetic diversity and to construct the fingerprints of Robinia pseudoacacia clones selected in Shandong province, which can lay a solid foundation for R. pseudoacacia new varieties selection, breeding and identification, and also provide a scientific basis for the conservation, assessment and utilization of R. pseudoacacia in other provinces of China.[Method]Fluorescent SSR primers were used for PCR amplification. The products were detected with capillary electrophoresis. The results were used to analyze genetic diversity and to construct fingerprints of 49 R. pseudoacacia clones.[Result]A total of 51 alleles were identified using 8 pairs of SSR primers, with a mean of 6.375 alleles per locus, ranging from 2-15. The mean value of PIC was 0.509 8, ranging from 0.092-0.879. Clustering analysis conducted with the method of average linkage between groups showed that clones ‘Luci 8’ and ‘Luci 13’ from Linyi, ‘Luci 40’ and ‘Luci 42’ from Qingdao, ‘Luci 10’ and ‘Luci 86’ from Rizhao displayed the closest genetic relationship, respectively. The 49 clones were not clustered together strictly in accordance with geographic areas. There were no obvious correlations between grouping and current growing regions of the clones. A total of 9 primer pairs were used for clonal identification, two of them namely Rply109 and rops16 can distinguish 45 clones, indicating an identification rate of 91.84%. Three alleles at one or two loci were detected in 22 clones, suggesting that these clones may be natural polyploidies.[Conclusion]R. pseudoacacia in Shandong province has relatively high genetic diversity. There were not obvious correlations between grouping and current growing regions of the clones. The two primers Rply109 and rops16 were determined to be efficient SSR markers for fingerprints construction and molecular identification of R. pseudoacacia clones, which can distinguish a proportion of 91.84% of the total number of clones. The fingerprints constructed with SSR markers can provide a basis for germplasm resources management, variety identification and intellectual property rights protection, it also provides a scientific basis for introduction, genetic improvement and breeding of R. pseudoacacia.

Key words: Robinia pseudoacacia, SSR, molecular marker, genetic diversity, fingerprint, cluster analysis

中图分类号:

毛秀红, 郑勇奇, 孙百友, 张元帅, 韩丛聪, 位晓, 荀守华. 基于SSR的刺槐无性系遗传多样性分析和指纹图谱构建[J]. 林业科学, 2017, 53(10): 80-89.

Mao Xiuhong, Zheng Yongqi, Sun Baiyou, Zhang Yuanshuai, Han Congcong, Wei Xiao, Xun Shouhua. Genetic Diversity and Fingerprints of Robinia pseudoacacia Clones Based on SSR Markers[J]. Scientia Silvae Sinicae, 2017, 53(10): 80-89.

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