中国农业科学 ›› 2020, Vol. 53 ›› Issue (9): 1756-1772.doi: 10.3864/j.issn.0578-1752.2020.09.006

• 专题:限制性两阶段多位点全基因组关联分析法的应用 • 上一篇    下一篇

大豆重组自交系群体异黄酮含量QTL连锁定位与关联定位的比较研究

刘再东,孟珊,贺建波(),邢光南,王吴彬,赵团结,盖钧镒()   

  1. 南京农业大学大豆研究所/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室/作物遗传与种质创新国家重点实验室/江苏省现代作物生产协同创新中心,南京 210095
  • 收稿日期:2019-09-09 接受日期:2020-01-02 出版日期:2020-05-01 发布日期:2020-05-13
  • 通讯作者: 贺建波,盖钧镒
  • 作者简介:刘再东,E-mail:2714699171@qq.com。|孟珊,E-mail:mshan84@163.com。
  • 基金资助:
    国家自然科学基金(31701447);国家作物育种重点研发计划(2017YFD0101500);国家作物育种重点研发计划(2017YFD0102002);长江学者和创新团队发展计划(PCSIRT_17R55);教育部111项目(B08025);中央高校基本科研业务费项目(KYT201801);农业部国家大豆产业技术体系CARS-04;江苏省优势学科建设工程专项;江苏省JCIC-MCP项目

A Comparative Study on Linkage and Association QTL Mapping for Seed Isoflavone Contents in a Recombinant Inbred Line Population of Soybean

ZaiDong LIU,Shan MENG,JianBo HE(),GuangNan XING,WuBin WANG,TuanJie ZHAO,JunYi GAI()   

  1. Soybean Research Institute, Nanjing Agricultural University/National Center for Soybean Improvement/Key Laboratory of Biology and Genetic Improvement of Soybean (General), Ministry of Agriculture/State Key Laboratory for Crop Genetics and Germplasm Enhancement/Jiangsu Collaborative Innovation Center for Modern Crop Production, Nanjing 210095
  • Received:2019-09-09 Accepted:2020-01-02 Online:2020-05-01 Published:2020-05-13
  • Contact: JianBo HE,JunYi GAI

摘要:

【目的】异黄酮是大豆等豆类植物中富含的一类次生代谢产物,对食品和保健产业有重要作用。大豆籽粒可分离出12种异黄酮组分,可归为三大类:大豆苷类异黄酮、染料木苷类异黄酮和黄豆苷类异黄酮。通过鉴定大豆籽粒异黄酮总含量及3个组分含量性状的加性及上位性QTL,进而全面解析其复杂的遗传构成。【方法】利用先进2号和赶泰2-2双亲衍生的大豆重组自交系群体NJRSXG,在5个环境下测定4个异黄酮含量性状:异黄酮总含量(total isoflavone content,SIFC)、大豆苷类异黄酮总含量(total daidzin group content,TDC)、染料木苷类异黄酮总含量(total genistin group content,TGC)和黄豆苷类异黄酮总含量(total glycitin group content,TGLC)。选用混合模型复合区间作图法(mixed-model-based composite interval mapping,MCIM)和限制性两阶段多位点全基因组关联分析方法(restricted two-stage multi-locus genome-wide association analysis,RTM-GWAS)进行异黄酮含量QTL检测。【结果】2个亲本在4个异黄酮含量性状上均存在较大差异,重组自交系群体异黄酮含量在高值、低值2个方向上均出现超亲分离,低值方向分离趋势强于高值方向。利用连锁定位MCIM方法共检测到4个异黄酮含量性状的19个加性QTL和16对上位性QTL,分布于15条染色体上。第14染色体重要标记区间GNE186b—Sat020内检测到3个新加性QTL:qSifc-14-1qTdc-14-2qTgc-14-1,且表型变异解释率最高。利用关联定位RTM-GWAS方法分别检测到4个异黄酮含量性状的51、66、42和36个关联标记位点,表型变异解释率为39.7%—52.5%,检测到的位点中覆盖了MCIM方法检测的19个加性QTL中的11个以及11个上位性QTL。候选基因分析分别在加性QTL区域和上位性QTL区域检测到93和100个候选基因,富集分析显示在第14染色体重要标记区间GNE186b—Satt020内,Glyma14g33227Glyma14g33244Glyma14g33715的功能与异黄酮代谢有关。【结论】连锁定位和关联定位2种方法结合能相对全面地检测异黄酮含量QTL。与连锁定位方法MCIM相比,关联定位方法RTM-GWAS检测的QTL更多,总遗传贡献率更高,但尚不能检测上位性QTL,2种方法定位结果可相互验证补充,大豆籽粒异黄酮含量由大量QTL/基因控制。

关键词: 大豆, 重组自交系群体, 异黄酮, 连锁作图, 关联定位, QTL

Abstract:

【Objective】Isoflavones are a group of phenolic secondary metabolites which are relatively abundant in soybean and some other legumes, and are important for food and healthcare industry. A total of 12 kinds of components are isolated from soybean seed, and can be grouped into three categories: daidzin group, genistin group and glycitin group. To understand the complex genetic constitutions of isoflavone content in soybean, the additive and epistatic quantitative trait loci (QTLs) conferring the total isoflavone content and its component contents were detected in the present study. 【Method】The NJRSXG recombinant inbred line (RIL) population derived from Xianjin 2 and Gantai-2-2 were used in this study. Four isoflavone content traits, i.e. the total seed isoflavone content (SIFC), the total daidzin group content (TDC), the total genistin group content (TGC) and the total glycitin group content (TGLC) were tested in five environments. The mixed model composite interval mapping (MCIM) and restricted two-stage multi-locus genome-wide association analysis (RTM-GWAS) were used for QTL detection. 【Result】There was a large difference in isoflavone content between the two parental lines of NJRSXG population, and transgressive segregations were observed in NJRSXG population while the transgressive trend in low isoflavone content direction were stronger than that in high isoflavone content direction. A total of 19 additive QTLs and 16 pairs of epistatic QTLs for the four isoflavone traits on 15 chromosomes were detected by MCIM. Three novel additive QTLs, i.e. qSifc-14-1, qTdc-14-2 and qTgc-14-1, were detected in the same important marker interval GNE186b-Sat020 on chromosome 14, and explained the highest phenotypic variation. A total of 51, 66, 42 and 36 significantly associated markers were detected by RTM-GWAS for SIFC, TDC, TGC and TGLC, respectively. The phenotypic variation explained by these markers was ranged from 39.7% to 52.5%, covering 11 additive QTLs and 11 epistatic QTLs detected by MCIM. Furthermore, a total of 93 and 100 candidate genes were annotated in the additive and epistatic QTL regions, respectively. Gene enrichment analysis indicated that three genes located in the important marker interval GNE186b-Satt020 on chromosome 14, i.e. Glyma14g33227, Glyma14g33244 and Glyma14g33715, were related to isoflavone metabolism. 【Conclusion】A relatively thorough detection of isoflavone content QTLs was achieved by using linkage and association mapping. Compared with the linkage mapping method MCIM, the association mapping method RTM-GWAS can detect more QTLs with larger total contribution to phenotypic variation, but cannot detect epistatic QTLs as in MCIM. The QTLs detected from the two methods can be used for complementary verification from each other. A large number of QTLs/genes are involved in the seed isoflavone contents of soybean.

Key words: soybean [Glycine max (L.) Merr.], isoflavone, genetic dissection, linkage mapping, association mapping, QTL