cDNA Cloning, Bioinformatics, and Expression Analysis of ApsANS in Acer pseudosieboldianum

Anthocyanin synthetase (ANS), a key enzyme in the final step of the anthocyanin synthesis pathway, catalyzes the conversion of leucoanthocyanidins to anthocyanins. In this study, an ANS structural protein (TRINITY_DN18024_c0_g1) was found to be associated with anthocyanin accumulation in <i>Acer pseudosieboldianum</i> leaves, named <i>ApsANS</i>. Real-time quantitative fluorescence PCR analysis revealed that the expression of <i>ApsANS</i> was significantly higher in red-leaved (variant) than green-leaved (wild-type) strains, which was consistent with the transcriptome data. The UPLC results showed that the cyanidin metabolites may be the key substance influencing the final color formation of <i>Acer pseudosieboldianum</i>. The <i>ApsANS</i> gene was cloned and analyzed through bioinformatics analysis. <i>ApsANS</i> has a total length of 1371 bp, and it encodes 360 amino acids. Analysis of the structural domain of the <i>ApsANS</i> protein revealed that <i>ApsANS</i> contains a PcbC functional domain. Protein secondary structure predictions indicate that α-helix, irregularly coiled, and extended chains are the major building blocks. Subcellular localization predicted that <i>ApsANS</i> might be localized in the nucleus. The phylogenetic tree revealed that <i>ApsANS</i> is relatively closely related to <i>ApANS</i> in <i>Acer palmatum</i>. The prediction of miRNA showed that the <i>ApsANS</i> gene is regulated by miR6200. This study provides a theoretical reference for further analyzing the regulatory mechanism of leaf color formation in <i>Acer pseudosieboldianum</i>.