中国农业科学 ›› 2023, Vol. 56 ›› Issue (1): 17-30.doi: 10.3864/j.issn.0578-1752.2023.01.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

干旱胁迫下甘蓝型油菜籽粒含油量和蛋白质含量变化的全基因组关联分析

胡盛(),李阳阳,唐章林,李加纳,曲存民,刘列钊()   

  1. 西南大学农学与生物科技学院,重庆400715
  • 收稿日期:2022-08-22 接受日期:2022-10-21 出版日期:2023-01-01 发布日期:2023-01-17
  • 通讯作者: 刘列钊
  • 作者简介:胡盛,E-mail:916340758@qq.com
  • 基金资助:
    重庆市研究生科研创新项目(CYS21122);国家自然科学基金(31971902);国家自然科学基金(32001509)

Genome-Wide Association Analysis of the Changes in Oil Content and Protein Content Under Drought Stress in Brassica napus L.

HU Sheng(),LI YangYang,TANG ZhangLin,LI JiaNa,QU CunMin,LIU LieZhao()   

  1. College of Agronomy and Biotechnology, Southwest University, Chongqing 400715
  • Received:2022-08-22 Accepted:2022-10-21 Online:2023-01-01 Published:2023-01-17
  • Contact: LieZhao LIU

摘要:

【目的】干旱是甘蓝型油菜生长发育过程中一种常见的非生物胁迫,严重影响了其产量和品质。联合全基因组关联分析和转录组差异表达分析,筛选干旱胁迫条件下影响甘蓝型油菜籽粒含油量和蛋白质含量变化的候选基因,为解释干旱胁迫下甘蓝型油菜籽粒含油量及蛋白质含量的变化提供理论基础。【方法】利用旱棚盆栽方式模拟干旱胁迫环境,使用183份甘蓝型油菜构成的自然群体,于2019和2020年在模拟干旱胁迫条件下收获2年的籽粒,并进行籽粒含油量以及蛋白质含量的测定,将得到的表型数据与60K芯片的基因型数据(包含34 103个SNP)进行全基因组关联分析,同时,结合相同处理下花后不同干旱时间段(30、40和50 d)的籽粒转录组差异基因数据,筛选共有基因,并利用甘蓝型油菜和拟南芥数据库注释信息、已报道的相关文献和转录组差异基因表达水平鉴定与干旱胁迫下甘蓝型油菜籽粒含油量及蛋白质含量变化相关的候选基因。【结果】通过对2年间籽粒含油量和蛋白质含量的分析,发现材料重复性较好,干旱胁迫造成甘蓝型油菜籽粒的含油量下降,蛋白质含量上升;全基因组关联分析得出的最佳模型主要为一般线性模型下的Q或naïve模型,共检测出38个显著关联位点(P<1/31597或P<1/31278),主要分布在A03(6个)、A04(8个)和C03(8个)染色体,各位点分别对含油量和蛋白质含量变化的贡献率均超过10%,其中含油量变化位点最大贡献率为23.97%,蛋白质含量变化位点最大贡献率为22.21%。通过整合全基因组关联分析和转录组分析,筛选出256个共有基因,根据基因注释和文献报道鉴定出与干旱胁迫下含油量及蛋白质含量变化相关的25个候选基因,其中包含转录因子(如bZIP transcription factor GBF6、TALE transcription factor ATH1、MYB-like Domain transcription factor MYBD、NAC transcription factor ANAC029和ERF transcription factor ERF111)、相关激酶(如与油脂相关的蛋白激酶CIPK9、水分胁迫响应激酶PIP5K1和代谢相关激酶PFK7)、相关蛋白(如与油脂相关转运蛋白ABCA9、与蛋白相关的存储蛋白CRU3、胁迫响应蛋白HUP26和M10、叶绿体蛋白DG238和CP12)等,涉及生长发育、光合反应、物质运输等多个生物学过程。【结论】鉴定出25个相关候选基因,可能响应干旱胁迫,并影响油菜籽粒蛋白质和含油量的积累。

关键词: 甘蓝型油菜, 干旱胁迫, 含油量, 蛋白质含量, 候选基因

Abstract:

【Objective】 Drought is one of the most adverse abiotic stresses that hinder growth and development of Brassica napus L., threatening its yield and quality. In this study, we identified the candidate genes that influence oil and protein content changes according to the results of the genome wide association mapping (GWAS) and differentially expressed genes (DEGs) under drought stress in B. napus. 【Method】 A natural population comprising 183 B. napus lines under drought stress across two years was genotyped with a Brassica 60K Illumina Infinium SNP array to perform GWAS. Combined with drought stressed seed transcriptome data from different treatment periods, the candidate genes related to the changes in oil and protein contents under drought stress were identified. 【Result】 The analysis showed that the best model for phenotypic data analysis in 2 years was mainly the Q or naïve model, and a total of 38 significant associated loci were detected (P<1/31597 or P<1/31278). By integrating GWAS and transcriptome results, 256 common genes were selected, and by comparing the database information of B. napus and Arabidopsis thaliana, 25 related candidate genes were identified. These genes, including transcription factors (e.g., bZIP transcription factor GBF6, TALE transcription factor ATH1, MYB-like Domain transcription factor MYBD, NAC transcription factor ANAC029, ERF transcription factor ERF111), related kinases (e.g., PIP5K1, PFK7), related proteins (e.g., seed stores protein CRU3, chloroplast protein DG238 and CP12, stress-responsive proteins HUP26 and M10), are involved in multiple biological processes such as photosynthetic reaction, substance transport, and stress response. For instance, oil-related protein kinase CIPK9, ABCA9 transporter, storage-related protein CRU3, may provide basis for explaining the changes in oil and protein contents of B. napus under drought stress. 【Conclusion】 The 25 identified candidate genes may affect the accumulation of protein and oil contents under drought stress during the seed development in B. napus.

Key words: Brassica napus L., candidate genes, drought stress, oil content, protein content