中国农业科学 ›› 2019, Vol. 52 ›› Issue (24): 4453-4469.doi: 10.3864/j.issn.0578-1752.2019.24.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

基于SSR标记的谷子主要农艺性状关联位点检测及等位变异分析

李剑峰1,张博1,全建章2,王永芳2,张小梅1,赵渊1,袁玺垒1,贾小平1(),董志平2()   

  1. 1 河南科技大学农学院,河南洛阳 471023
    2 河北省农林科学院谷子研究所/国家谷子改良中心,石家庄 050035
  • 收稿日期:2019-05-28 接受日期:2019-06-25 出版日期:2019-12-16 发布日期:2020-01-15
  • 通讯作者: 贾小平,董志平
  • 作者简介:李剑峰,E-mail:1299569468@qq.com。
  • 基金资助:
    国家自然科学基金(31471569);国家“十二五”科技支撑计划(2011BAD06B01-1)

Associated Loci Detection and Elite Allelic Variations Analysis of Main Agronomic Traits in Foxtail Millet (Setaria italica L.) Based on SSR Markers

JianFeng LI1,Bo ZHANG1,JianZhang QUAN2,YongFang WANG2,XiaoMei ZHANG1,Yuan ZHAO1,XiLei YUAN1,XiaoPing JIA1(),ZhiPing DONG2()   

  1. 1 College of Agriculture, Henan University of Science and Technology, Luoyang 471023, Henan
    2 Institute of Millet, Hebei Academy of Agriculture and Forestry Sciences/National Millet Improvement Center, Shijiazhuang 050035
  • Received:2019-05-28 Accepted:2019-06-25 Online:2019-12-16 Published:2020-01-15
  • Contact: XiaoPing JIA,ZhiPing DONG

摘要:

【目的】通过在海南省乐东县、河南省洛阳市、吉林省吉林市和公主岭市4个不同地理环境调查谷子10个主要性状,进行SSR标记与性状的关联分析,获得单一环境特异表达位点及多个环境共同表达位点,发掘优异等位变异,探究谷子可能的生态适应性形成机制,为开展谷子分子辅助选择育种奠定基础。【方法】连续2年在吉林省吉林市和公主岭市、河南省洛阳市、海南省乐东县调查102份谷子资源株高、穗长、叶片数、穗粗、抽穗期、穗码数、码粒数、穗重、穗粒重、千粒重10个主要农艺性状的基础上,首先利用SPSS19.0软件对每个地理环境进行性状间相关性分析,再用70个多态性SSR标记对谷子资源进行基因型鉴定,分析102份谷子资源的遗传多样性和群体遗传结构,最后用TASSEL5.0软件进行标记间的连锁不平衡分析,并用GLM和MLM 2种模型开展分子标记与表型性状的关联分析。【结果】除了千粒重,其余9个性状间在4个地理环境普遍存在显著或极显著正相关,千粒重只在在吉林市、公主岭市、洛阳市3个环境与穗粒重、株高、穗长、穗重呈显著或极显著正相关,在海南乐东县与其他9个性状没有显著相关性;70对SSR引物共检测到397个等位基因,平均每个标记的观测等位基因数、有效等位基因数、期望杂合度、Shannon指数分别为6、2.24、0.4637、0.7738;遗传多样性和群体结构分析均将102份谷子材料分为4个群,来自河南的谷子材料散布在4个群中,表现较丰富的遗传多样性;70对SSR标记间连锁不平衡分析未发现明显的连锁不平衡结构;GLM、MLM 2种模型共检测到10个关联标记,结合等位效应分析确定b115、MPGC13、b227、b194、p56在吉林市、公主岭市2个环境与穗重、穗长、叶片数、抽穗期4个性状关联,sigms9034、b125在洛阳市与穗长、穗码数关联,P18和p59在乐东县与穗重关联,p6在吉林市、公主岭市和洛阳市分别与叶片数和穗码数关联,单个标记对表型变异的平均贡献率在7.76%—34.05%;3个标记等位基因sigms9034-168、P18-166、p56-244分别能够显著增加穗长、穗重,缩短抽穗期。【结论】连续2年获得了5个在吉林市、公主岭市稳定表达的标记位点b115、MPGC13、b227、b194和p56,2个在洛阳市稳定表达的标记位点sigms9034、b125,2个在乐东县稳定表达的位点P18和p59,1个在洛阳市和吉林市、公主岭市同时稳定表达的标记位点p6;获得了3个可用于穗部性状、生育期标记辅助选择的优异等位基因sigms9034(168 bp)、P18(166 bp)、p56(244 bp)。

关键词: 谷子, SSR, 地理环境, 农艺性状, 关联分析

Abstract:

【Objective】Through the investigation of 10 major traits in foxtail millet at Ledong, Hainan province, Luoyang, Henan province, Jilin and Gongzhuling, Jilin province, totally four different geographical environments, association analysis between SSR markers and the ten traits was performed to obtain loci expressed in single environment or multiple environments, excavate elite allelic variations, explore the probable mechanism forming ecological adaptation and provide foundation for launching molecular-assistant selection breeding in foxtail millet.【Method】Based on the survey of ten traits (plant height, panicle length, number of leaves, panicle diameter, heading stage, spikelet number, grain number per branch, spike weight, grain weight per panicle and 1000-grain weight) from 102 foxtail millet varieties at Jilin and Gongzhuling, Jilin province, Luoyang, Henan province and Ledong, Hainan province for two consecutive years, correlation analysis of ten traits at each geographical environment was first performed by SPSS 19.0 software, then the 102 foxtail millet varieties were genotyped by 70 polymorphic SSR markers, and further genetic diversity and population genetic structure of these varieties were analyzed. Finally, linkage disequilibrium analysis among markers and association analysis between molecular markers and phenotypic traits were carried out by GLM and MLM models of TASSEL 5.0 software. 【Result】There existed significant or very significant positive correlations among most of the nine agronomic traits except 1000-grain weight across four geographical environments. Only significant or very significant positive correlations were found between 1000-grain weight and grain weight per panicle, plant height, panicle length, spike weight at Jilin, Gongzhuling and Luoyang, no significant correlations were found between 1000-grain weight and other nine traits at Ledong environment. Totally 397 alleles were detected in 70 pairs of SSR primers, giving average observed allele number, effective allele number, expected heterozygosity, Shannon index of 6, 2.24, 0.4637 and 0.7738 per marker respectively. Both genetic diversity analysis and population structure analysis divided 102 foxtail millet materials into 4 groups, and the varieties from Henan province scattered in each of the four groups, showing more abundant genetic diversity. Linkage disequilibrium analysis showed that no obvious LD structures were found among 70 SSR markers. Totally 10 associated markers were detected by GLM and MLM models, combined with allele effect analysis results, it could be determined that b115, MPGC13, b227, b194 and p56 were associated with spike weight, panicle length, number of leaves and heading stage at Jilin, Gongzhuling, sigms9034 and b125 were associated with panicle length and spikelet number at Luoyang, P18 and p59 were associated with spike weight at Ledong, p6 was associated with number of leaves and spikelet number at Jilin, Gongzhuling and Luoyang respectively. The average contribution rates of single marker to phenotypic variation ranged from 7.76% to 34.05%. Three alleles, sigms9034-168, P18-166 and p56-244, could increase panicle length and spike weight, shorten heading stage significantly, which would be used to improve panicle traits and shorten growth period by marker-assisted selection breeding.【Conclusion】Five marker loci (b115, MPGC13, b227, b194 and p56) were steadily detected at Jilin and Gongzhuling, two marker loci (sigms9034, b125), two marker loci (P18, p59) were steadily detected at Luoyang and Ledong respectively for two consecutive years. One marker locus (p6) was steadily detected at Jilin, Gongzhuling and Luoyang, three geographical environments for two consecutive years. Three elite alleles (sigms9034-168bp, P18-166bp, p56-244bp) that could be used to carry out marker-assisted selection for panicle traits and growth period were obtained.

Key words: foxtail millet, SSR, geographical environment, agronomic traits, correlation analysis