基于ISSR标记的山茱萸核心种质构建

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摘要山茱萸(Cornus officinalis)是我国独有的木本中药材，良种培育对提高山茱萸产业效益有重要意义，种质资源是良种培育的基础。为了更加高效地开展山茱萸种质资源收集评价和保存利用，本研究从自全国11个省(市)采集129份山茱萸样品，利用11条简单重复序列区间(inter-simple sequence repeat, ISSR)引物进行PCR扩增，使用随机取样和位点优先取样两种聚类分析策略，分别采用简单匹配(simple matching, SM)遗传相似系数、Jaccard遗传相似系数和Nei & Li遗传相似系数3种遗传距离，进行核心种质的选择。用SPSS 19.0软件对原种质、核心种质和保留种质的群体遗传参数进行T检验，以评价核心种质的代表性和异质性，对原种质和核心种质的形态指标进行异质性检验，来确认核心种质的代表性。11条引物共从129个样品扩增出87个条带，全部为多态性位点，群体观测等位基因数(number of alleles, Na)为2.000 0，平均有效等位基因数(number of effective alleles, Ne)为1.438 8，平均Nei's遗传多样性指数(Nei's genetic diversity index, H)为0.259 3，平均香农信息指数(average Shannon's information index, I)为0.400 6，表明收集的山茱萸材料在DNA分子水平上有较高的遗传多样性。聚类分析的结果，认为不同地区的居群间种质资源存在一定的地理隔离和遗传分化。根据6种聚类取样方法结果的比较，采用位点优先取样策略和Nei & Li遗传距离的S1D3组，选取出的34份样品，是6组初选种质中群体遗传多样性参数最高的，有效等位基因数、Nei's遗传多样性指数和香农信息指数均显著大于保留种质，遗传多样性较保留种质丰富。对比原种质和核心种质的叶片和果实的13个表型指标，除叶片宽、叶柄长和果干重3个性状有差异外，其他10个性状指标核心种质与原种质均无显著差异，所选的核心种质能够从表型指标上代表原种质资源。本研究从来自全国11个省(市)的129份山茱萸材料中抽取出34份，构建的核心种质，包含了原种质26.36%的样本。通过对核心种质和原种质、保留种质的群体遗传多样性参数进行比较，结合对13个表型性状的数据进行确认，构建的核心种质可以代表原种质的遗传多样性，研究的结果可以为山茱萸种质资源收集保存和新品种培育作参考。

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	赵宁
	杨孟莉
	李继东
	郑先波
	谭彬
	叶霞
	赵志浩
	冯建灿

关键词 ：山茱萸, 核心种质, ISSR标记, 位点优先策略, 随机取样策略

Abstract：Germplasm resources are foundation for superior cultivar breeding, and essential for improve the economic efficiency for Cornus officinalis, a special herb tree species in China. The current study select C. officinalis core collection in the samples from all over the country by inter-simple sequence repeat (ISSR) markers, for the collection, evaluation, preservation and usage of germplasm. One hundred and twenty nine samples were collected from 11 provinces all over China. Total DNA were extracted by modified cetrimonium trimethyl ammonium bromide (CTAB) method and amplified by 11 ISSR markers. To select the core construction, cluster sampling adopts 2 strategies: Random sampling and site sampling, while genetic distances were estimated in 3 ways: Similar matching (SM) genetic similarity coefficient, Jaccard genetic similarity coefficient or Nei & Li genetic similarity coefficient. T test was carried out on the genetic parameters of the original collection, core collection and germplasm collection, to evaluate the representative and heterogeneity. The heterogeneity of the original germplasm and core collection was tested to confirm the representative of the core collection. Eleven primers amplified 87 bands from 129 samples, all of which were polymorphic, and the average loci number of each primer was 7.91. For the whole 129 samples, the number of alleles (Na) was 2.000 0, the average effective number of alleles (Ne) was 1.438 8, average Nei's genetic diversity index (H) was 0.259 3, the average Shannon's information index (I) was 0.400 6. These parameters showed the samples had high genetic diversity at the DNA level. Cluster analysis showed that geographical isolation and genetic differentiation existed in the populations from different areas. Cluster sampling results under 2 sampling strategies and 3 types of genetic distance were compared, the S1D3 group selected 34 samples by site preferred sampling strategy and Nei & Li genetic distance clustering showed the highest genetic diversity parameters. The number of effective alleles, Nei 's genetic diversity index and Shannon information index of the S1D3 group were significantly higher than the initial collection in the 0.01 level of probability, and the genetic diversity was more abundant than the reserve germplasm. Thirteen phenotypic indicators of the original germplasm and the core collection were compared, except for leaf width, petiole length and dried fruit weight differed at the 0.05 level, other 10 traits had no significant difference with the original germplasm. The phenotypic indicators of the selected core germplasm could represent the original germplasm resources. A C. officinalis core collection including 34 samples were selected from 129 germplasms collected from 11 provinces all over China, contained 26.36% of the original germplasm. By comparing the genetic diversity parameters of the core collection，original germplasm and the reserve germplasm, along with the confirmation by 13 phenotypic traits data, the core collection can represent the genetic diversity of the original germplasm. The result can be used in C. officinalis germplasm resource conservation and superior cultivar breeding.

Key words： Cornus officinalis Core collection ISSR markers Allele preferred sampling strategy Random sampling strategy

收稿日期: 2016-10-09 出版日期: 2017-03-31

ZTFLH:

S727.3

基金资助:河南省高校科技创新团队支持计划;林业公益性行业科研专项

通讯作者: 冯建灿 E-mail: jcfeng@henau.edu.cn

引用本文:

赵宁杨孟莉李继东郑先波谭彬叶霞赵志浩冯建灿. 基于ISSR标记的山茱萸核心种质构建[J]. , 2017, 25(4): 579-587.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I4/579

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