【Objective】Cellulose synthase A (CesA) represents the core catalytic enzyme orchestrating cellulose biosynthesis in plants, directly governing cellulose accumulation and playing pivotal roles in wood formation, cell wall development, and lignification processes. Dalbergia odorifera, a rare and endemic timber species in China, holds significant ornamental, economic, and medicinal value. However, excessive logging has led to a drastic decline in its wild germplasm resources, pushing the species toward endangerment. Molecular breeding offers an efficient strategy to enhance wood yield and quality, making it imperative to elucidate the functions of key genes regulating wood development for genetic improvement. This study undertakes a genome-wide identification and systematic bioinformatic analysis of the CesA gene family in D. odorifera to delineate the structural characteristics, evolutionary relationships, and expression patterns of DoCesA genes. The findings aim to establish a theoretical foundation for wood trait improvement and elite cultivar breeding, while also advancing understanding of cellulose synthesis mechanisms in this non-model species. 【Method】In this study, comprehensive bioinformatics approaches were employed to systematically identify members of the DoCesA gene family. The investigation encompassed detailed characterizations including chromosomal localization mapping, physicochemical property profiling of encoded proteins, subcellular compartmentalization prediction, and structural feature analyses. Specifically, we conducted rigorous examinations of conserved motifs, functional domains, gene architectures, and transmembrane helix configurations. Promoter region dissection further revealed a repertoire of cis-regulatory elements potentially governing transcriptional regulation. To elucidate evolutionary dynamics, phylogenetic reconstruction and interspecies synteny analysis were performed, providing insights into lineage-specific expansion patterns and orthologous relationships. Expression pattern characterization and alternative splicing analysis were achieved through integration of high-throughput transcriptome sequencing data with experimental validation using quantitative real-time PCR (qRT-PCR). 【Result】Fifteen non-redundant DoCesA genes (DoCesA1-DoCesA15) were identified, exhibiting uneven distribution across eight chromosomes. The deduced proteins range from 976 to 1,311 amino acids, with average molecular weight of 121.12 kDa, theoretical pI values of 5.75-7.6, instability indices of 36.45-42.69, and aliphatic indices of 80.00-89.56. All DoCesA proteins were predicted to localize to the plasma membrane. Conserved motif analysis revealed ten shared motifs, while domain scans identified two signature CesA domains (Cellulose_synt and zf-UDP) in each isoform. Transmembrane helix predictions indicated six C-terminal and two N-terminal helices in most proteins. Promoter regions contained abundant cis-elements responsive to light, phytohormones, and abiotic stresses. Phylogenetic analysis classified the family into two subgroups: Group I (DoCesA8, DoCesA11-15) associated with primary cell wall synthesis, and Group II (DoCesA2-7, 9) linked to secondary cell wall biosynthesis. Synteny analysis identified 13, 27, 3, and 20 orthologous pairs between D. odorifera and A. thaliana, G. max, O. sativa, and P. trichocarpa, respectively. Transcriptome profiling demonstrated that primary-wall-related DoCesA genes showed higher expression in phloem, whereas secondary-wall-associated copies peaked in sapwood and xylem transitional zones. qRT-PCR confirmed DoCesA2 and DoCesA4 exhibited maximal transcriptional abundance in transitional zones. Alternative splicing analysis revealed seven DoCesA genes undergo four splicing variants: alternative 3′/5′ splice sites, intron retention, exon skipping, and mutually exclusive exons. Notably, DoCesA12 generated 12 distinct transcripts with tissue-biased expression: DoCesA12.4 (sapwood), DoCesA12.7/11 (leaves), DoCesA12.8/12 (transitional zone), and others enriched in phloem. 【Conclusion】This study provides the first genome-wide identification and systematic characterization of the DoCesA gene family in D. odorifera, comprehensively clarifying their structural features, evolutionary relationships, and expression dynamics. The integration of transcriptome analysis and qRT-PCR validation deepens understanding of DoCesA genes' roles in wood development and alternative splicing regulation. These results not only enrich CesA gene research in D. odorifera but also offer critical insights for functional genomics studies aimed at improving wood traits through molecular breeding strategies. Future work should focus on validating specific DoCesA genes' functions using overexpression or CRISPR/Cas9 systems, particularly those involved in secondary cell wall biosynthesis, to accelerate genetic improvement of this valuable species.
Key words
Dalbergia odorifera /
CesA family /
structural characteristics /
phylogenetic relationship /
expression pattern /
alternative splicing
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