A bHLH gene NnTT8 of Nelumbo nucifera regulates anthocyanin biosynthesis

Lotus is an important aquatic ornamental plant, whose flower color is one of the key horticultural traits that determines its ornamental value. Previous studies revealed that anthocyanins largely determined the red color of lotus flower, which are also the main component that has beneficial effects on human health. However, the regulation mechanism of flower pigmentation in lotus flower remains unclear. In the present study, in order to further understand the regulatory mechanism underlying the anthocyanin biosynthesis, a bHLH gene NnTT8 was characterized to be phylogenetically close to AtTT8 and the bHLH proteins from other plant species that have been indicated to be involved in the positive regulation of anthocyanin biosynthesis. Complementation analysis in Arabidopsis tt8 mutant showed that NnTT8 could function similarly to AtTT8 in regulating anthocyanin and proanthocyanin biosynthesis. An MYB transcription factor capable of interacting with NnTT8 was also characterized from lotus. The identification of a bHLH transcription factor playing regulatory roles in anthocyanin biosynthesis is crucial, as it might help to obtain more in-depth insight into the coloration of lotus and help in breeding high anthocyanin content lotus variety that can be explored for lotus flower beverages.