School of Life Sciences, Research Center of Buckwheat Industry Technology, Guizhou Normal University, Guiyang 550025, China.
Int J Mol Sci. 2023 Dec 11;24(24):17368. doi: 10.3390/ijms242417368.
Tartary buckwheat () is an important plant, utilized for both medicine and food. It has become a current research hotspot due to its rich content of flavonoids, which are beneficial for human health. Anthocyanins (ATs) and proanthocyanidins (PAs) are the two main kinds of flavonoid compounds in Tartary buckwheat, which participate in the pigmentation of some tissue as well as rendering resistance to many biotic and abiotic stresses. Additionally, Tartary buckwheat anthocyanins and PAs have many health benefits for humans and the plant itself. However, little is known about the regulation mechanism of the biosynthesis of anthocyanin and PA in Tartary buckwheat. In the present study, a bHLH transcription factor (TF) FtTT8 was characterized to be homologous with AtTT8 and phylogenetically close to bHLH proteins from other plant species. Subcellular location and yeast two-hybrid assays suggested that FtTT8 locates in the nucleus and plays a role as a transcription factor. Complementation analysis in mutant showed that FtTT8 could not recover anthocyanin deficiency but could promote PAs accumulation. Overexpression of in red-flowering tobacco showed that inhibits anthocyanin biosynthesis and accelerates proanthocyanidin biosynthesis. QRT-PCR and yeast one-hybrid assay revealed that FtTT8 might bind to the promoter of and suppress its expression, while binding to the promoter of and upregulating its expression in K326 tobacco. This displayed the bidirectional regulating function of FtTT8 that negatively regulates anthocyanin biosynthesis and positively regulates proanthocyanidin biosynthesis. The results provide new insights on TT8 in Tartary buckwheat, which is inconsistent with TT8 from other plant species, and FtTT8 might be a high-quality gene resource for Tartary buckwheat breeding.
苦荞麦是一种重要的植物,兼具药用和食用价值。由于其富含对人体健康有益的类黄酮,因此成为当前的研究热点。矢车菊素(ATs)和原花青素(PAs)是苦荞麦中两种主要的类黄酮化合物,它们参与某些组织的着色,并赋予植物对多种生物和非生物胁迫的抗性。此外,苦荞麦花色苷和 PAs 对人类和植物本身都有许多健康益处。然而,人们对苦荞麦中花色苷和 PA 生物合成的调控机制知之甚少。本研究中,鉴定了一个 bHLH 转录因子(TF)FtTT8,其与 AtTT8 同源,且在系统发育上与其他植物物种的 bHLH 蛋白亲缘关系较近。亚细胞定位和酵母双杂交实验表明,FtTT8 定位于细胞核,作为转录因子发挥作用。在 突变体中的互补分析表明,FtTT8 不能恢复花色苷缺陷,但可以促进 PAs 积累。在红花烟草中的过表达表明, 抑制花色苷生物合成并加速原花青素生物合成。QRT-PCR 和酵母单杂交实验表明,FtTT8 可能与 启动子结合并抑制其表达,同时与 启动子结合并上调 K326 烟草中的表达。这显示了 FtTT8 的双向调控功能,即负调控花色苷生物合成,正调控原花青素生物合成。这些结果为苦荞麦中的 TT8 提供了新的见解,与其他植物物种中的 TT8 不一致,并且 FtTT8 可能是苦荞麦育种的优质基因资源。