College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
Int J Mol Sci. 2022 Feb 12;23(4):2033. doi: 10.3390/ijms23042033.
Apples () are rich in flavonols, and 5-aminolevulinic acid (ALA) plays an important role in the regulation of plant flavonoid metabolism. To date, the underlying mechanism of ALA promoting flavonol accumulation is unclear. Flavonol synthase (FLS) is a key enzyme in flavonol biosynthesis. In this study, we found that ALA could enhance the promoter activity of in the 'Fuji' apple and improve its expression. With as bait, we screened a novel transcription factor MdSCL8 by the Yeast One-Hybrid (Y1H) system from the apple cDNA library which we previously constructed. Using luciferase reporter assay and transient GUS activity assay, we verified that MdSCL8 inhibits the activity of promoter and hinders expression, thus reducing flavonol accumulation in apple. ALA significantly inhibited expression. Therefore, ALA promoted the expression of and the consequent flavonol accumulation probably by down-regulating MdSCL8. We also found that ALA significantly enhanced the gene expression of MdMYB22 and MdHY5, two positive regulators of . We further demonstrated that MdMYB22 interacts with MdHY5, but neither of them interacts with MdSCL8. Taken together, our data suggest MdSCL8 as a novel regulator of and provide important insights into mechanisms of ALA-induced flavonol accumulation in apples.
苹果富含类黄酮,而 5-氨基乙酰丙酸(ALA)在植物类黄酮代谢的调节中起着重要作用。迄今为止,ALA 促进类黄酮积累的潜在机制尚不清楚。类黄酮合酶(FLS)是类黄酮生物合成的关键酶。在本研究中,我们发现 ALA 可以增强‘富士’苹果中 的启动子活性并提高其表达。我们以前构建了一个苹果 cDNA 文库,利用该文库,通过酵母单杂交(Y1H)系统作为诱饵筛选到一个新的转录因子 MdSCL8。通过荧光素酶报告基因检测和瞬时 GUS 活性检测,我们验证了 MdSCL8 抑制 启动子的活性并阻碍 表达,从而减少苹果中的类黄酮积累。ALA 显著抑制 表达。因此,ALA 通过下调 MdSCL8 促进 表达,进而促进类黄酮积累。我们还发现 ALA 显著增强了 正向调控因子 MdMYB22 和 MdHY5 的基因表达。我们进一步证明 MdMYB22 与 MdHY5 相互作用,但它们都不与 MdSCL8 相互作用。综上所述,我们的数据表明 MdSCL8 是 调控因子,为 ALA 诱导苹果中类黄酮积累的机制提供了重要的见解。