基于T2T基因组鉴定大豆R2R3-MYB基因家族及干旱和盐胁迫下的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0745

摘要/Abstract

摘要：

目的 R2R3-MYB转录因子在植物响应逆境胁迫过程中发挥着重要作用，旨在对大豆R2R3-MYB转录因子进行完整鉴定以及研究大豆R2R3-MYB基因在逆境胁迫中的作用。方法利用大豆无gap端粒到端粒（T2T）基因组对大豆R2R3-MYB家族基因进行系统鉴定和进化分析，并使用RT-qPCR对该家族的基因在干旱和盐胁迫下进行表达分析。结果大豆基因组中包含324个R2R3-MYB基因，新鉴定到77个，分布在20条染色体上。进化分析结果显示大豆R2R3-MYB家族基因可分为4个亚组，共线性基因对在进化树的分支上距离更近。通过启动子分析发现，与ABA和抗旱相关的顺式作用元件最多。通过表达分析，推测10个基因参与了大豆耐盐过程，其中GmMYB19和GmMYB76已被报道，GmMYB89和GmMYB214是首次鉴定并发现可能与大豆抗旱耐盐性有关。结论在大豆中鉴定到321个R2R3-MYB家族基因，通过干旱和盐胁迫诱导的表达模式分析确定了10个抗旱耐盐的候选基因。

关键词: 大豆, R2R3-MYB基因家族, 生物信息学分析, 干旱和盐胁迫, 表达分析, T2T

Abstract:

Objective R2R3-MYB transcription factors play an important role in plant response to stress conditions. It is aimed to fully identify the R2R3-MYB transcription factors in soybean and understand the role of soybean R2R3-MYB genes in stress responses. Method The soybean R2R3-MYB family genes were systematically identified and evolutionarily analyzed using the soybean Telomere-to-Telomere (T2T) genome. Real-time fluorescence quantitative PCR (RT-qPCR) was used to analyze the expression of this family of genes under drought and salt stress conditions. Result The soybean genome contains 324 R2R3-MYB genes, 77 of which were newly identified and distributed on 20 chromosomes. The results of evolutionary analysis showed that soybean R2R3-MYB family genes can be divided into four subgroups, and the colinear gene pairs were closer on the branches of the evolutionary tree. Promoter analysis revealed that most cis-acting elements related to ABA and drought resistance were present. Through expression analysis, 10 genes were speculated to be involved in the process of soybean salt tolerance, of which GmMYB19 and GmMYB76 have been reported, and GmMYB89 and GmMYB214 were identified for the first time and found to be related to soybean drought and salt tolerance. Conclusion A total of 321 R2R3-MYB family genes were identified in soybean, and 10 candidate genes for drought and salt tolerance were identified through analysis of their expression patterns under drought and salt stress conditions.

Key words: Glycine max, R2R3-MYB gene family, bioinformatics analysis, drought and salt stress, expression analysis, T2T

李志强, 罗正乾, 徐琳黎, 周国慧, 屈丝雨, 刘恩良, 顼东婷. 基于T2T基因组鉴定大豆R2R3-MYB基因家族及干旱和盐胁迫下的表达分析[J]. 生物技术通报, 2025, 41(5): 141-152.

LI Zhi-qiang, LUO Zheng-qian, XU Lin-li, ZHOU Guo-hui, QU Si-yu, LIU En-liang, XU Dong-ting. Identification of the Soybean R2R3-MYB Gene Family Based on the T2T Genome and Their Expression Analysis under Drought and Salt Stress[J]. Biotechnology Bulletin, 2025, 41(5): 141-152.

图/表 9

表1 使用的引物信息

Table 1 Primer information used

引物名称 Primer name	正向引物 Forward sequence （5'-3'）	反向引物 Reverse sequence （5'-3'）
Gm_MYB114	GGACCATACCTCCTTGGCAG	GGTGATGGTGGTGTGTCCAA
Gm_MYB137	AGCTGCCAGGAAGAACAGAC	ACGCAACGTGTCTCTGAACT
Gm_MYB169	TCCTCTGATGGAAGCTGCAC	TGGCTAAACCACTCACCAGC
Gm_MYB19	ATCGGTGGGCCAAAATAGCA	TTAGCCTGCTTCACCACTCG
Gm_MYB208	CCCAGTGACACACAGTCCTC	TACTGGTGCATGCCAAGGAG
Gm_MYB214	TGGTTCAACAGGAAGTGCGT	GCTTGAGGCCAGGATGTAGG
Gm_MYB249	GGCATGTGATGATGGCAAGG	GTGACACAAGGGTGGCTGTA
Gm_MYB51	GGCAAAGGACATGGATCGGA	AACCAACCTCCGTAACGCTT
Gm_MYB76	GCCAGGCAGAACAGACAATG	GGCATAGGAGTTGCCTGGTT
Gm_MYB89	GATGGTCTACCATTGCGGCT	GTGTGGTGTGCCAATGGTTC
GmActin	CGGTGGTTCTATCTTGGCATC	GTCTTTCGCTTCAATAACCCTA

图1 大豆R2R3-MYB基因的染色体位置红色代表已鉴定的，黑色代表新鉴定的

Fig. 1 Chromosome location of soybean R2R3-MYB geneRed indicates those that have been identified, and black indicates those newly identified

图2 大豆和拟南芥R2R3-MYB基因的系统进化树不同的颜色表示不同的分组，红色星星代表大豆R2R3-MYB基因，绿色星星代表拟南芥R2R3-MYB基因

Fig. 2 Phylogenetic tree of soybean and Arabidopsis thalianaR2R3-MYB genesDifferent colors indicate different groups. The red star indicates the soybean R2R3-MYB gene, and the green star indicates the Arabidopsis thalianaR2R3-MYB gene

图3 大豆R2R3-MYB基因的共线性分析

Fig. 3 Collinearity analysis of soybean R2R3-MYB genes

图4 R2R3-MYB基因家族基因结构和motif基序分析

Fig. 4 Gene structure and motif analysis of the R2R3-MYB gene family

图5 大豆R2R3-MYB基因家族启动子顺式作用元件分析A：不同类别启动子顺式作用元件数量；B：逆境胁迫相关顺式作用元件分类；C：生长发育相关顺式作用元件分类；D：激素相关顺式作用元件分类

Fig. 5 Analysis of cis-acting elements in the promoters of soybean R2R3-MYB gene familyA: Number of cis-acting elements in different categories of promoters. B: cis-acting elements related to stress. C: cis-acting elements related to growth and development. D: cis-acting elements related to hormones

图6 大豆R2R3-MYB基因家族基因干旱和盐胁迫下的表达分析A：干旱胁迫下的表达量热图；B：盐胁迫下的表达量热图；C：干旱和盐胁迫下的共有和特异的差异表达R2R3-MYB基因数量

Fig. 6 Expression analysis of genes in soybean R2R3-MYB gene family under drought and salt stressA: Expression calorimetry under drought stress. B: Expression calorimetry under salt stress. C: Number of common and specific differentially expressed R2R3-MYB genes under drought and salt stress

图7 10个候选基因在干旱胁迫的表达模式结果以平均值±SD（n=3，*P<0.05，**P<0.01），下同

Fig. 7 Expression patterns of 10 candidate genes under drought stressThe results are presented as means±SD (n=3, *P<0.05, **P<0.01), the same below

图8 10个候选基因在盐胁迫的表达模式

Fig. 8 Expression patterns of 10 candidate genes under salt stress

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