Identification and Expression Analysis of CAD Gene Family in Soybean（Glycine max （L.） Merr.）

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

Abstract

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

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.

Figures/Tables 8

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

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

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

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

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

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

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

Fig. 8 The CAD genes expression under drought stress

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