Buddleja alternifolia Exhibits Extensive CCD4 Gene Expansion and Diversification for Crocetin Biosynthesis
A recent study has revealed that the plant *Buddleja alternifolia* possesses a significantly expanded family of Carotenoid Cleavage Dioxygenase 4 (CCD4) enzymes, with 12 identified members, making it one of the largest CCD4 subgroups in the Lamiales order. This expansion is primarily driven by recent tandem duplication events, particularly on chromosome 10, with evidence of ongoing birth-and-death evolution, including the presence of pseudogenes. The research, which involved genomic, phylogenetic, transcriptional, structural analyses, and functional characterization of 11 *BaCCD4* paralogs, demonstrated extensive biochemical diversification among these enzymes. While some paralogs showed no detectable activity or produced asymmetric carotenoid cleavage products like citraurin, two specific enzymes, KAG8367281 and KAG8375220, were identified as capable of cleaving zeaxanthin to generate crocetin dialdehyde, a direct precursor of crocetin and crocins. These crocetin-producing enzymes belong to closely relate...