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林业科学 ›› 2016, Vol. 52 ›› Issue (4): 48-58.doi: 10.11707/j.1001-7488.20160406

• 论文与研究报告 • 上一篇    下一篇

基于SRAP分子标记的苦楝种质资源遗传多样性分析

廖柏勇1, 王芳1, 陈丽君1, 刘明骞1, 欧阳昆唏1, 阙青敏1, 惠文凯2, 李培2, 陈晓阳1   

  1. 1. 华南农业大学林学与风景园林学院 广东省森林植物种质创新与利用重点实验室 亚热带农业生物资源保护与利用国家重点实验室 广州 510642;
    2. 北京林业大学生物科学与技术学院 北京 100083
  • 收稿日期:2015-05-04 修回日期:2015-06-10 出版日期:2016-04-25 发布日期:2016-05-05
  • 通讯作者: 陈晓阳
  • 基金资助:
    广东省林业科技创新项目(2011KJCX002)。

Genetic Diversity of Germplasm Resources of Melia azedarach Revealed by SRAP Markers

Liao Boyong1, Wang Fang1, Chen Lijun1, Liu Mingqian1, Ouyang Kunxi1, Que Qingmin1, Xi Wenkai2, Li Pei2, Chen Xiaoyang1   

  1. 1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources Guangdong Provincial Key Laboratory of Forest Plant Germplasm Innovation and Utilization College of Forestry and Landscape Architecture, South China Agricultural University Guangzhou 510642;
    2. College of Biological Sciences and Technology, Beijing Forestry University Beijing 100083
  • Received:2015-05-04 Revised:2015-06-10 Online:2016-04-25 Published:2016-05-05

摘要: [目的] 利用相关序列扩增多态性(SRAP)分子标记对来自17个省(区)的31个苦楝种源进行遗传多样性分析,为苦楝种质资源的保存和育种计划的制订提供依据。[方法] 通过SRAP分子标记获得1/0数据矩阵,计算SRAP分子标记各项遗传参数,同时进行分子方差分析(AMOVA)。随后计算遗传距离矩阵并进行主坐标分析(PCoA)、邻接法聚类分析(Neighbour-Joining)以及遗传距离和地理距离Mantel相关性分析。[结果] 从783对SRAP引物中筛选出的20对引物可扩增出257条清晰的条带,其中145条具有多态性。引物多态性信息指数(PIC)为0.262~0.478,均值为0.385。PIC值较高的10对引物,可扩增出56条特异多态带,可以作为快速鉴别苦楝种源的工具。AMOVA结果显示,38.96%的遗传变异来源于种源间,61.04%的遗传变异来源于种源内。山区的贵州册亨和黎平,云南勐腊和麻栗坡,湖南东安、炎陵和浏阳,四川达州,河北保定,安徽滁州种源的遗传多样性高于其他种源。邻接聚类法根据Nei's遗传距离将31个种源分为东西2类,西部云南、四川、贵州和甘肃的8个种源构成类群Ⅰ,其他地区的23个种源构成东部类群Ⅱ。类群Ⅱ中北方的济南、保定、泰安、渭南和许昌5个种源聚为一小类,华南地区的屯昌、五指山、钦州和仁化种源聚为一小类,其他相邻和相近的种源大多聚在一起。PCoA分析的种源间双标图结果与邻接法聚类结果基本一致。种源内双标图显示相同种源内的单株大都聚在一起,且种源间和种源内双标图总体分布大体一致。Mantel检验结果表明,种源间遗传距离和地理距离相关性显著(r=0.256,P=0.003)。[结论] SRAP分子标记可以准确、有效地用于苦楝遗传多样性分析。苦楝遗传变异主要来源于种源内,而种源间基因交流有限。种源遗传多样性整体偏低,而部分山区种源遗传多样性较高。在选择高遗传多样性种源的基础上,苦楝选育应侧重种源内家系和单株选择。苦楝种源特征明显,地理环境差异对其遗传多样性具有一定的影响。31个种源大致可分为东西2类,东部种源从北到南有多个聚类中心,其种质资源应采用多点就地保存方式。我国苦楝遗传多样性起源中心或许存在于2类不同种源的生态过渡区。

关键词: 苦楝, SRAP, 遗传多样性, 亲缘关系

Abstract: [Objective] This study was aimed to analyze genetic diversity of 31 provenances of Melia azedarach from 17 provinces in China by Sequence Related Amplified Polymorphism(SRAP) molecular markers, and to provide a basis for conservation of germplasm resources and breeding of M. azedarach. [Method] We obtained SRAP 1/0 matrix via SRAP molecular markers, calculated the parameters of SRAP molecular markers, and conducted analysis of molecular variance(AMOVA). Then we computed the genetic distance matrix of SRAP data, and carried out principal coordinate analysis(PCoA), Neighbour-Joining cluster analysis and Mantel's test for the correlation between genetic distance matrix and geographic distance matrix.[Result] 20 pairs of primer combinations, screened from 783 pairs of primer combinations, amplified 257 clear bands, of which 145 were polymorphic. The polymorphism information content(PIC) values ranged from 0.262 to 0.478, with an average value of 0.385. The screened 10 primer combinations with the highest PIC values could distinguish almost all of the populations, so that they could be used to identify provenances of M. azedarach rapidly. The results of analysis of molecular variance(AMOVA)indicated that 38.96% genetic variation is derived from among populations and 61.04% is derived from within populations. Mountain provenances GZ2 and GZ3 from Guizhou, YN1 and YN2 from Yunnan, HUN1, HUN2 and HUN3 from Hunan, SC2 from Sichuan, HEB from Hebei, and AH from Anhui displayed higher genetic diversity than other provenances did. The Neighbour-Joining cluster analysis and principal coordinates analysis displayed similar results, 31 M. azedarach provenances were clustered into east and west groups. Provenances of Cluster Ⅰ were distributed along the west of China, including provenances from Yunnan, Guizhou, Sichuan and Gansu, and cluster Ⅱ contained other 23 provenances that were distributed in the east of China. Within the cluster Ⅱ, SD1, HEB, SD2, SX, and GS provenances from north area were clustered into sub-group Ⅱ 1. The four southern provenances, HAN2, HAN1, GX2, and GD2, were clustered into sub-group Ⅱ 4. The rest provenances either neighbouring or close to each other were mostly clustered together. According to the Mantel test, there was a significant correlation between genetic distance and geographic distance(r=0.256, P=0.003).[Conclusion] SRAP markers can be accurately and effectively used in genetic diversity analysis of M. azedarach. More genetic variation of M. azedarach existed within provenances, while gene exchange among provenances was limited. The overall genetic diversity of the provenances was relatively low, while some mountain provenances had higher genetic diversity. On the basis of selecting provenances with high genetic diversity, breeding of M. azedarach should be focused on selection of families and individuals within provenance. Different provenances had their own obvious characteristics, and differences in geographical environment had a certain impact on their genetic diversity. The 31 provenances were clustered into the east and the west groups, and the east groups had several diversity centres, so it was suggested that the germplasm resources be conserved in multiple sites in situ respectively. The centre of the origin of genetic diversity of M. azedarach may have an ecological transition zone between the two different groups of provenances in China.

Key words: Melia azedarach, SRAP, genetic diversity, genetic relationship

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