中国农业科学 ›› 2015, Vol. 48 ›› Issue (16): 3121-3131.doi: 10.3864/j.issn.0578-1752.2015.16.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

糜子骨干种质遗传多样性和遗传结构分析

董俊丽1,2,王海岗1,陈凌1,王君杰1,曹晓宁1,王纶1,乔治军1   

  1. 1山西省农业科学院农作物品种资源研究所/农业部黄土高原作物基因资源与种质创制重点实验室/杂粮种质资源发掘与遗传改良山西省重点实验室,太原 030031
    2山西大学,太原030006
  • 收稿日期:2015-02-12 出版日期:2015-08-16 发布日期:2015-08-16
  • 通讯作者: 乔治军,E-mail:nkypzs@126.com
  • 作者简介:董俊丽,E-mail:296998342@qq.com。王海岗,E-mail:nkywhg@126.com。董俊丽和王海岗为同等贡献作者。
  • 基金资助:
    国家现代农业产业技术体系建设专项(CARS-07-12.5-A12)

Analysis of Genetic Diversity and Structure of Proso Millet Core Germplasm

DONG Jun-li1,2, WANG Hai-gang1, CHEN Ling1, WANG Jun-jie1, CAO Xiao-ning1, WANG Lun1, QIAO Zhi-jun1   

  1. 1Institute of Crop Germplasm Resources of Shanxi Academy of Agricultural Sciences/Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture/Shanxi Key Laboratory of Genetic Resources and Genetic Improvement of Minor Crops, Taiyuan 030031
    2Shanxi University, Taiyuan 030006
  • Received:2015-02-12 Online:2015-08-16 Published:2015-08-16

摘要: 【目的】糜子生育期短、耐干旱、耐瘠薄、水分利用效率高,了解糜子资源的遗传多样性和遗传结构,为今后糜子杂交育种、种质创新、挖掘抗旱基因及资源的高效利用提供理论依据。【方法】采用表型鉴定和SSR分子标记对糜子资源进行遗传多样性检测。利用模糊隶属函数法分析糜子种质的株高、主穗长、叶片长、叶片宽、主茎节数、主茎粗、单株穗重、单株粒重和千粒重9个表型性状的分布情况。利用DPS7.05软件进行表型性状的遗传多样性分析、相关性分析和主成分分析,综合评价糜子种质资源的优劣。利用CTAB法提取糜子嫩叶基因组DNA,并利用SSR分子标记技术对不同地区的96份糜子种质资源的基因组DNA进行PCR扩增,后经8%聚丙烯酰胺凝胶电泳分离,银染后显色。利用PowerMaker 3.25软件计算每对引物的等位基因数(A)、主要等位基因频率(M)、基因多样性指数(He)和多态性信息含量指数(PIC),并进行N-J遗传距离的统计分析;利用Structure 2.3.1分析群体遗传结构。【结果】糜子表型遗传多样性分析表明:9个表型性状分布集中,且绝大部分呈极显著相关;单株粒重和单株穗重遗传变异最丰富,不同省份的资源在表型性状上表现出不同的遗传多样性,山西资源表型遗传多样性最丰富。采用主成分分析法和综合评价法表明,内糜1号的综合性状表现最差,宁糜15号的综合性状表现最好。采用19对SSR引物对96份糜子种质资源进行遗传多样性分析,共检测出112个等位变异;每个位点的等位变异数为3—9个,平均5.9个;平均主要等位基因频率为0.7045;平均基因多样性指数为0.4097;平均多态性信息含量位点百分数为39.2%。不同地理来源糜子种质资源的遗传多样性分析表明,各省份间糜子资源的亲缘关系均较近;山西省资源的基因多样性指数及多态性信息含量百分数最高,分别为0.357和33.01%。基于模型的遗传结构分析和基于遗传距离的聚类分析将试验材料划分为3个类群,两种分类结果有一定相似性,皆与生态环境密切相关。【结论】糜子遗传变异较为丰富,遗传多样性高,尤其是山西糜子资源的遗传多样性最丰富;不同地理来源的糜子种质资源亲缘关系均较近,且其遗传多样性与生态环境密切相关。

关键词: 糜子, 综合评价, 群体遗传结构, 聚类分析, 遗传多样性

Abstract: 【Objective】 The objective of this study is to understand the genetic diversity of proso millet germplasm from different regions and to provide an important theoretical foundation for crossbreeding, excavating drought genes and germplasm creation for future, because of its characters, such as short growth period, bearing drought and barren land. 【Method】Genetic diversity detection was done for proso millet resources by phenotypic identification and SSR markers. Distribution of nine phenotypic traits, including plant height, main spike length, leaf length, blade width, number of main stem section, the main stem diameter, spike weight per plant, grain weight per plant and thousand kernel weight were analyzed by subordinate function. Genetic diversity analysis, correlation analysis and principal component analysis of phenotypic traits were done by software DPS7.05; comprehensive evaluation on proso millet germplasm. Good genomic DNA was extracted from young leaves of 96 proso millet germplasm resources from different areas following the CTAB method, and then were amplified by simple sequence repeat molecular markers to analyze genetic diversity and genetic structure. Separation of the amplified fragments was performed on 8% denaturing polyacrylamide gels. The gels were stained with AgNO3 for visualizing the SSR fragments. The No. of allele (A), Major allele frequency(M), Gene diversity(He), polymorphism information content(PIC) were estimated by software PowerMaker 3.25. Neighbor-joined cluster analysis was aslo carried by software PowerMaker 3.25. Population genetic structure was analyzed by software Structure 2.3.1.【Result】Through genetic diversity analysis of the nine phenotypes of proso millet, it was known that the spike weight and grain weight per plant of genetic variation were the most abundant. Phenotypic traits were more concentrated and most of which were apparently interrelated. Genetic diversity of phenotypic traits of germplasm from Shanxi was the richest. Genetic diversity of phenotypic traits was different in different provinces. Principal component analysis and comprehensive evaluation were done for 9 phenotypic traits of 96 copies of proso millet germplasm resources. Comprehensive evaluation value of Neimi No.1 was the lowest, while that of Ningmi No.15 was the highest. Genetic diversity of 96 copies of proso millet germplasm resources was detected by 19 pairs of SSR primers and 112 allelic variation genes were found. The number of alleles per locus ranged from 3 to 9 and the average was 5.9, the average major allele frequency was 0.7045, the average gene diversity index was 0.4097, and the average percentage of polymorphism information content sites was 39.2%. Genetic diversity analysis of germplasm from different regions showed a close relationship among the regions. Genetic diversity index and polymorphic information content percentage of proso millet germplasm resources in Shanxi were 0.357 and 33.01%, respectively. Experimental materials were divided into three groups by genetic structure analysis based on modules and cluster analysis based on genetic distance. Results of these two division methods have similarity. Resources in three groups were connected with geographical environment closely.【Conclusion】 Proso millet germplasm resources were analyzed systematically from aspects of phenotype and molecule. The results showed that proso millet is rich in genetic variation and has a high diversity. What’s more, genetic diversity of proso millet germplasm resources in Shanxi is the most abundant.

Key words: proso millet, comprehensive evaluation, population structure, cluster analysis, genetic diversity