Maturation of litchi (Litchi chinensis) fruit is characterized by dramatic changes in pigments in the pericarp and flavor compounds in the aril. Among them, the biosynthesis of anthocyanins is most noticeable. Previous studies showed that LcMYB1 and LcbHLH transcription factors participated in regulating the anthocyanin biosynthesis in litchi. However, the roles of other MYB factors remain unclear. In this study, we cloned and characterized the function of LcMYB5, a novel R2R3-MYB identified from litchi transcriptome. Although LcMYB5 was constitutively expressed in litchi tissues and its expressions was not correlated with tissue coloration, overexpression of LcMYB5 resulted in enhanced biosynthesis of anthocyanins in tobacco and petunia concurrent with the up-regulation of their endogenous bHLHs and key structural genes in anthocyanin precursor biosynthesis. These results indicate that LcMYB5 is an R2R3 transcriptional factor regulates anthocyanin biosynthesis either by directly activating the expression of key structural genes such as DFR or by indirectly up regulating the expressions of endogenous bHLH regulators. More interestingly, the pH values in petals and leaves from transgenic lines were significant lower than those in both untransformed tobacco and petunia, indicating LcMYB5 is also associated with pH regulation. The expressions of LcMYB5 and its bHLH partner LcbHLH1 were consistent with the expression of putative tissue acidification gene LcPH1, and the changes in malic acid provided further evidence for the close relationship between LcMYB5 and tissue acidification. Taking together, our study indicated that LcMYB5 is involved in not only anthocyanin biosynthesis but also tissue acidification.

Genome-wide Identification and Expression Analyses of PAL Genes in Different Color Radish

Litchi (Litchi chinensis Sonn.) is a popular subtropical fruit with a red pericarp that is primarily determined by the accumulation of anthocyanins. The peel color and fruit quality are also influenced by proanthocyanins (PAs), which play roles in fruit development and postharvest quality. In this study, we identified LcMYB2 as a key regulator of both anthocyanin and PA biosynthesis in litchi. Phylogenetic analysis revealed that LcMYB2 belongs to the VvMYB5 subclade. Expression analysis showed that LcMYB2 is highly expressed in the early stages of fruit development. Its expression pattern was consistent with that of LcLAR and LcANR, two key genes in the PA biosynthetic pathway. Subcellular localization and protein–protein interaction assays confirmed that LcMYB2 localizes to the nucleus and interacts with LcbHLH3. Dual-luciferase reporter assays demonstrated that the LcMYB2-LcbHLH3 complex activates the promoters of LcLAR and LcANR, supporting its role in regulating PA biosynthesis. Furthermore, overexpression of LcMYB2 in tobacco resulted in the synthesis of anthocyanins and PAs in the flower, indicating that LcMYB2 can regulate anthocyanin and PA biosynthesis. Additionally, transgenic tobacco plants with LcMYB2 overexpression exhibited delayed anther dehiscence, suggesting a broader role in plant development. These findings highlight the multifunctional nature of LcMYB2 in regulating both anthocyanin and PA biosynthesis, as well as its involvement in reproductive development.