大豆CAD基因家族的鉴定及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0928

摘要/Abstract

摘要：

目的研究大豆肉桂醇脱氢酶（Cinnamyl Alcohol Dehydrogenase, CAD）基因家族（GmCADs）成员特征和在不同胁迫下的表达模式，为后续深入解析GmCADs基因家族生物学功能奠定基础。方法基于大豆基因组数据，利用生物信息学方法对GmCADs基因家族成员进行鉴定，对其编码蛋白质的特征、系统发育关系、基因结构、保守基序等进行分析。结果在大豆全基因组水平鉴定了18个CAD家族基因（GmCAD1-GmCAD18），分布在13条染色体上，氨基酸数量在219-364 aa之间。系统发育分析显示，GmCADs基因家族分为4个亚族，同一亚族基因具有相似的基因结构和保守基序。共线性分析表明片段复制是GmCADs基因家族扩张的主要形式。选择压力分析表明，GmCADs基因处于纯化选择。GmCADs基因家族的启动子上含有丰富的与大豆光响应、激素响应、逆境响应和生长发育过程相关的顺式作用元件。通过5个间接蛋白，所有GmCADs基因形成了一个复杂的蛋白质相互作用网络，且GO功能分析显著富集在CAD活性和木质素代谢过程等条目。GmCADs成员在不同大豆组织和逆境下（干旱、盐、冷、荫蔽和高温）表达具有一定的选择性，其中GmCAD11和GmCAD18表达量较高。RT-qPCR验证表明，在盐和干旱处理后，9个GmCADs基因表达量在不同时间显著改变，主要表现为上调表达。结论 18个大豆CAD成员在分布、结构及功能上存在多样性，且GmCADs基因可能在大豆植株生长发育过程中响应非生物胁迫。

关键词: 大豆, 肉桂醇脱氢酶, 基因家族, 蛋白互作网络, 表达分析

Abstract:

Objective The characteristics and expression analysis under different stresses of the Cinnamyl Alcohol Dehydrogenase (CAD) gene family (GmCAD) in soybean were studied, providing a theoretical basis for further research on the biological function of the GmCADs gene family. Method Based on the soybean genome data, the CAD gene family members were screened and identified using bioinformatics methods. The characteristics of the encoded proteins, phylogenetic relationships, gene structure and conserved motifs were analyzed. Result A total of 18 GmCADs (GmCAD1-GmCAD18) were identified in the whole soybean genome, which were unevenly distributed on 13 chromosomes, encoding 219 to 364 amino acids. Phylogenetic analysis revealed that the GmCADs gene family can be divided into four subfamilies, and the genes within the same subfamily share similar gene structures and conserved motifs. The collinearity analysis indicates that fragment duplication was the main form of expansion in the GmCADs gene family. Selection pressure analysis showed that the GmCADs gene was under purifying selection. The promoters of the GmCADs gene family containing abundant cis-acting elements were mainly associated with the light response, hormone response, stress response and growth development processes. The protein interaction network prediction results showed a complex protein interaction network, which was formed by all GmCADs genes through five indirect proteins, and GO functional analysis significantly enriched in terms such as CAD activity and lignin metabolism process. The GmCAD members demonstratecertain selectivity in expression under different tissues and stress conditions (such as drought, salt, cold, shade, and high temperature stress). RT-qPCR validation indicates the expressions of nine GmCAD significantly changed at different time points under salt and drought treatments, mainly showing upregulation. Conclusion The distribution, structure and function of the 18 soybean CAD members are diverse, and the GmCADs gene may respond to abiotic stress during the growth and development of soybean plants.

Key words: soybean, CAD, gene family, protein interaction network, expression analysis

苏燕竹, 李达, 张爱爱, 刘永光, 张秀荣, 薛其勤. 大豆CAD基因家族的鉴定及表达分析[J]. 生物技术通报, 2026, 42(4): 101-113.

SU Yan-zhu, LI Da, ZHANG Ai-ai, LIU Yong-guang, ZHANG Xiu-rong, XUE Qi-qin. Identification and Expression Analysis of CAD Gene Family in Soybean（Glycine max （L.） Merr.）[J]. Biotechnology Bulletin, 2026, 42(4): 101-113.

图/表 8

图1 GmCADs基因在大豆染色体上的分布（A）和系统进化分析（B）A：染色体上的颜色表示基因的密度，颜色越红，密度越高，颜色越蓝，密度越低

Fig. 1 Chromosomal distribution (A) and phylogenetic analysis (B) of GmCAD genesA: The colors on the chromosome indicate the density of the gene. The redder the color, the higher the density. The bluer the color, the lower the density

图2 GmCADs基因结构（A）和保守基序（B）

Fig. 2 Gene structure (A) and putative conserved motifs (B) analysis of GmCAD genes

图3 CAD基因家族共线性分析A：大豆CAD家族成员种内共线性分析。红色曲线连接片段复制的基因对；外侧部分指示基因在染色体上的对应位置，绿色框对应不同的染色体（Chr01-Chr20）；红色锯齿线突出显示了每条染色体上的基因密度高低；图的内侧框进一步强调了染色体内的基因密度，图例表示基因密度，黄色到红色表示基因密度由小变大。B：大豆与不同物种间CAD基因的共线性分析。红色高亮线为GmCADs基因的物种间共线性对，灰色线条为大豆与其他物种基因组的所有共线性区块

Fig. 3 Colinear analysis in the CAD gene familyA: Intraspecific colinear analysis of GmCAD genes family. Red curved line connects duplicated gene pairs. The outer part indicates the locations of these genes on the chromosome, and the green boxs correspond to different chromosomes (Chr 01-Chr 20). Red serrated lines highlight the gene density on each chromosome. The inner frames of the chart further emphasize the gene density in the chromosome, and with yellow to red indicating a change from small to large gene density. B: Colinear analysis of CAD gene between soybean and other species. The red lines highlight collinear pairs of GmCAD genes, while the gray lines in the background indicate collinear blocks between the genome of soybean and the other species

图4 GmCADs基因家族启动子的顺式作用元件分析

Fig. 4 Analysis of cis-acting elements in the promoter of GmCAD gene family

图5 GmCADs基因的蛋白质相互作用网络（A）和GO富集分析（B）BP：生物过程；CC：细胞组分；MF：分子功能

Fig. 5 Protein-protein interaction (PPI) network (A) and GO enrichment analysis (B) of GmCAD genesBP: Biological process. CC: Cellular component. MF: Molecular function

图6 大豆GmCADs家族基因组织表达特性（A）和不同胁迫条件下表达模式分析（B-F）B-F：分别是干旱、荫蔽、寒冷、高温和盐胁迫下GmCADs基因的表达模式。图中CK均表示对照，DT表示干旱胁迫，图C中“A”是耐荫大豆，“C”是不耐荫大豆，“T”是遮荫处理

Fig. 6 Tissue-specific expression characteristics (A) and expression patterns analysis under different stress conditions (B-F) of GmCAD family genes in soybean(Glycine max (L.) Merr.)B-F: the expression patterns of the GmCAD genes under conditions of drought (DT), shade, cold, heat and salt stress, respectively. In the figures, CK refers to the control, and DT to drought stress. In Figure C, ‘A’ refers to shade-tolerant soybeans, ‘C’ to shade-intolerant soybeans, and ‘T’ to shading treatment

图7 盐胁迫下CAD基因的表达量*和**分别表示盐胁迫处理后1-48 h与处理前0 h比较表达量具有显著（P<0.05）和极显著差异（P<0.01），下同

Fig. 7 The CAD genes expression under salt stress*and ** indicate that there are significant difference and extremely significant differences in expressions when comparing 1 h to 48 h after salt stress treatment with 0 before the treatment, with P<0.05 and P<0.01, the same below

图8 干旱胁迫下CAD基因的表达量

Fig. 8 The CAD genes expression under drought stress

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