中国农业科学 ›› 2025, Vol. 58 ›› Issue (1): 10-29.doi: 10.3864/j.issn.0578-1752.2025.01.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

大豆LOX基因家族的全基因组鉴定及GmLOX15A1基因等位变异对百粒重的影响

王伟(), 吴传磊, 胡晓渝, 李佳佳, 白鹏宇, 王郭伋, 苗龙, 王晓波*()   

  1. 安徽农业大学农学院,合肥 230036
  • 收稿日期:2024-05-23 接受日期:2024-06-11 出版日期:2025-01-01 发布日期:2025-01-07
  • 通信作者:
    王晓波,E-mail:
  • 联系方式: 王伟,E-mail:1275621348@qq.com。
  • 基金资助:
    国家重点研发计划(2021YFD1201605); 安徽省自然科学基金(2308085MC88)

Genome-Wide Identification of Soybean LOX Gene Family and the Effect of GmLOX15A1 Gene Allele on 100-Seed Weight

WANG Wei(), WU ChuanLei, HU XiaoYu, LI JiaJia, BAI PengYu, WANG GuoJi, MIAO Long, WANG XiaoBo*()   

  1. College of Agriculture, Anhui Agricultural University, Hefei 230036
  • Received:2024-05-23 Accepted:2024-06-11 Published:2025-01-01 Online:2025-01-07

摘要:

【目的】基于全基因组鉴定分析大豆LOX基因家族成员,了解各成员的分类进化关系,研究各基因成员在不同组织中的表达特异性以及对非生物胁迫的响应,为进一步研究LOX基因家族的分子特征、进化历程以及功能研究提供理论基础。【方法】基于Ensembl数据库中水稻和拟南芥物种LOX蛋白序列,在大豆全基因组数据库中BLASTP比对同源LOX蛋白序列,使用MEGA X软件构建系统进化树;采用网站MEME进行蛋白保守基序分析;运用在线软件GSDS 2.0分析基因结构;运用TBtools进行染色体定位绘制;用McscanX分析大豆LOX家族复制基因;利用PlantCARE网站预测大豆LOX基因家族启动子元件;通过TBtools绘制不同组织及非生物胁迫下大豆基因表达热图,并对与百粒重显著相关的优异等位变异位点GmLOX15A1-G/A进行分子标记开发。【结果】大豆中共鉴定到43个LOX基因,不均匀地分布在13条染色体上。共线性分析表明,GmLOX基因在进化过程中经历了广泛的复制。同时,在LOX基因启动子中检测到39种不同类型的顺式调控元件,表明它们可能参与了不同的生长发育、光反应、应激反应与激素诱导等途径。表达模式分析发现LOX基因在大豆不同组织中具有不同的表达程度,表明该家族成员具有组织和时空表达特异性。干旱胁迫条件下,GmLOX基因在大豆根系和叶片中表达差异显著(P<0.05),其中,GmLOX3A3GmLOX7A1GmLOX20B1GmLOX13A1GmLOX20A2在根系和叶片中均显著上调或下调表达,推测GmLOX基因可能在逆境胁迫响应中发挥重要作用。同时,发现GmLOX15A1在籽粒组织中高表达且在该基因编码区第七外显子存在一处G/A优异等位变异,对该变异位点开发分子标记,并利用来自不同生态区1 200份大豆种质资源2年的百粒重数据,分析GmLOX15A1不同单倍型与百粒重的相关性,结果表明,相较于GmLOX15A1-A基因型,携带GmLOX15A1-G等位基因大豆种质的平均百粒重提高2.33 g(P<0.001)。【结论】在大豆中共鉴定到43个LOX家族成员,可分为3个亚家族。GmLOX基因启动子区存在大量激素和胁迫响应的顺式作用元件,在干旱胁迫响应中发挥不同的作用。其中,GmLOX15A1在籽粒组织中高表达且该基因编码区第七外显子存在一处G/A优异等位变异,相较于GmLOX15A1-A基因型,携带GmLOX15A1-G等位基因大豆种质的平均百粒重显著提高2.33 g,该位点可作为大豆籽粒大小遗传改良的优异单倍型。

关键词: 大豆, 脂肪氧合酶, 生物信息学, 表达分析, 逆境响应, 百粒重

Abstract:

【Objective】Based on whole genome identification and analysis of soybean LOX gene family members, to understand the taxonomic evolutionary relationships of each member, to study the expression specificity of each gene member in different tissues and their response to abiotic stress, which provided a theoretical basis for further research on the molecular characteristics, evolutionary process, and function of the LOX gene family. 【Method】Based on the LOX protein sequences of rice and Arabidopsis species in Ensembl database, BLASTP alignment of homologous LOX protein sequences in soybean whole genome database was performed, and MEGA X software was used to construct a phylogenetic tree; Using website MEME for protein conserved motif analysis; Using online software GSDS 2.0 to analyze gene structure; Using TBtools for chromosome localization drawing; Analyze soybean LOX family replication genes using McscanX; Using the PlantCARE website to predict the promoter elements of soybean LOX gene family; Draw gene expression heatmaps of soybean under different tissues and abiotic stress using TBtools, and develop molecular markers for the excellent allele variant GmLOX15A1-G/A significantly correlated with 100-seed weight.【Result】A total of 43 LOX genes were identified in soybean, unevenly distributed on 13 chromosomes. Collinearity analysis indicates that the GmLOX gene has undergone extensive replication during the evolutionary process. Meanwhile, 39 different types of cis regulatory elements were detected in the LOX gene promoter, indicating that they may be involved in different pathways such as growth and development, light response, stress response, and hormone induction. Expression pattern analysis revealed that the LOX gene has different levels of expression in different tissues of soybean, indicating that members of this family have tissue and spatiotemporal expression specificity. Under drought stress conditions, the GmLOX gene was significantly differentially expressed in soybean roots and leaves (P<0.05). Among them, GmLOX3A3, GmLOX7A1, GmLOX20B1, GmLOX13A1, and GmLOX20A2 were significantly upregulated or downregulated in roots and leaves, suggesting that the GmLOX gene may play an important role in response to stress. At the same time, it was found that GmLOX15A1 is highly expressed in grain tissue and there is an excellent G/A allele variation in the seventh exon of the gene coding region. Molecular markers were developed for this variant site, and the correlation between different haplotypes of GmLOX15A1 and 100-seed weight was analyzed using 1 200 soybean germplasm resources from different ecological regions over a period of 2 years. The results showed that compared to the GmLOX15A1-A genotype, the average 100-seed weight of soybean germplasm carrying the GmLOX15A1-G allele gene increased by 2.33 g (P<0.001). 【Conclusion】A total of 43 members of the LOX family were identified in soybeans, which can be divided into 3 subfamilies. The promoter region of the GmLOX gene contains a large number of cis acting elements that respond to hormones and stress, playing different roles in drought stress response. Among them, GmLOX15A1 is highly expressed in grain tissue and there is an excellent G/A allele variation in the seventh exon of the coding region of this gene. Compared with the GmLOX15A1-A genotype, the average 100-seed weight of soybean germplasm carrying the GmLOX15A1-G allele gene is significantly increased by 2.33 g. This locus can be used as an excellent haplotype for genetic improvement of soybean grain size.

Key words: soybean, LOX lipoxygenase, bioinformatics, expression analysis, stress response, 100-seed weight