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The benzyl alcohol O-acetyl transferase, anthocyanin O-hydroxycinnamoyl transferase, N-hydroxycinnamoyl anthranilate benzoyl transferase, and deacetylvindoline 4-O-acetyltransferase (BAHD) enzymes play a critical role in regulating plant metabolites and affecting cell stability. In the present study, members of the  BAHD  gene family were recognized in the genome of  Theobroma cacao  and characterized using various bioinformatics tools. We found 27 non-redundant putative  tcBAHD  genes in cacao for the first time. Our findings indicate that  tcBAHD  genes are diverse based on sequence structure, physiochemical properties, and function. When analyzed with BAHDs of  Gossypium raimondii  and  Corchorus capsularis  clustered into four main groups. According to phylogenetic analysis,  BAHD  genes probably evolved drastically after their divergence. The divergence time of duplication events with purifying selection pressure was predicted to range from 1.82 to 15.50 MYA. Pocket analysis revealed that serine amino acid is more common in the binding site than other residuals, reflecting its key role in regulating the activity of tcBAHDs. Furthermore, cis-acting elements related to the responsiveness of stress and hormone, particularly ABA and MeJA, were frequently observed in the promoter region of  tcBAHD  genes. RNA-seq analysis further illustrated that  tcBAHD13  and  tcBAHD26  are involved in response to  Phytophthora megakarya  fungi. In conclusion, it is likely that evolutionary processes, such as duplication events, have caused high diversity in the structure and function of  tcBAHD  genes.

The BAHD Gene Family in Cacao (Theobroma cacao, Malvaceae): Genome-Wide Identification and Expression Analysis