The anthocyanin formation of purple leaf is associated with the activation of LfiHY5 and LfiMYB75 in crape myrtle

Purple-leafed plants not only have a higher resistance to biotic and abiotic stresses, but also have higher ornamental value. Anthocyanins are vital for leaf color formation, growth and development of purple leaves. However, the molecular mechanism underlying purple leaf formation in Lagerstroemia indica remains unclear. Metabolomic and transcriptomic analysis of purple-leafed cultivar ‘Ebony Embers’ and green-leafed cultivar ‘Arapahoe’ showed that the high expression of anthocyanin structure genes induced hyperaccumulation of cyanidin and pelargonidin derivatives, making the leaves purple. LfiHY5, LfiMYB75 and LfibHLH1 were identified using correlation analysis and weighted gene co-expression network analysis. In ‘Arapahoe’ × ‘Ebony Embers’ population, LfiHY5 and LfiMYB75 showed significant positive correlation with leaf anthocyanin content. Transient expression of LfiMYB75 and LfiHY5 in tobacco and purple-leafed crape myrtle indicated that the two genes activated anthocyanin synthesis. Yeast two-hybrid analysis showed that LfiMYB75 and LfibHLH1 could form a complex that enhanced anthocyanin synthesis. Yeast monohybrid and dual-luciferase assays confirmed that LfiHY5 activated the expression of LfiMYB75, to activate the transcription of anthocyanin structural genes LfiCHS and LfiANS. Moreover, there were three alleles of LfiHY5 in crape myrtle, and the different sequences had different activation effects on LfiMYB75. In conclusion, the results showed that LfiHY5 led to upregulate the transcription of LfiMYB75, and LfiMYB75 formed a complex with LfibHLH1, which increased the transcription level of LfiCHS and LfiANS to affect anthocyanin synthesis in crape myrtle.