The red coloration of the mango 'Irwin' skin is an important factor determining its value in the Japanese domestic luxury fruit market. In the present study, to investigate the molecular mechanism underlying anthocyanin biosynthesis of mango fruit skin, UFGT-like genes were isolated and the expression profile of anthocyanin-related genes was determined. Several UFGT-like genes were identified in transcriptome data of red 'Irwin' mango skin and two genes, MiUFGT1 and MiUFGT3, were considered to be involved in mango skin coloration. Deduced amino acid sequences of these genes exhibited high similarity to other plant UFGTs and contained the conserved PSPG box common to the glycosyltransferase family. The presence of a glutamine and a histidine residue at the C-terminus end of the PSPG box in MiUFGT1 and MiUFGT3, respectively, implied that MiUFGT1 and MiUFGT3 use glucose and galactose, respectively, as a sugar donor; however, the actual function and sugar donor preference of these enzymes remain to be elucidated. Expression analysis of anthocyanin-related genes during skin coloration suggested that MiCHS and MiANS, as well as MiUFGT1 and MiUFGT3, play important roles in the anthocyanin biosynthesis of mango fruit skin and that the expression of these genes is regulated by the MYB transcription factor, as reported in other plant species.
Red mangos (Mangifera indica L.) accumulate appreciable amounts of cyanidin-based anthocyanins in the skin, and previous studies showed that these anthocyanins contain galactose as a sugar moiety. To date, two UDP:flavonoid 3-O-glycosyltransferase (UFGT)-like genes named MiUFGT1 and MiUFGT3 (MiUFGalT3) have been isolated from mango 'Irwin' peel as anthocyanin-related UFGT genes, but the function of the proteins of the genes have not yet been elucidated. In this study, we characterized recombinant MiUFGT1 and MiUFGalT3 expressed in Escherichia coli. In the presence of quercetin as an acceptor, rMiUFGT1 showed marginal glucosylation activity, while rMiUFGalT3 exhibited significant galactosylation activity 20-fold higher than its glucosylation activity. Specificity analysis using purified MiUFGalT3 found that rMiUFGalT3 almost equally accepts anthocyanidins and flavonols. The anthocyanins extracted from the 'Irwin' skins were cyanidin 3-O-galactoside and 7-O-methylcyanidin 3-O-galactoside by instrumental analyses, which is consistent with previous results obtained for other red mango cultivars. The results suggest that MiUFGalT3 is responsible for the red coloration of 'Irwin' mango fruit skins.
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