State Key Laboratory of Tea Plant Biology and Utilization , Anhui Agricultural University , Hefei , Anhui 230036 , China.
College of Horticulture , Qingdao Agricultural University , Qingdao 266109 , China.
J Agric Food Chem. 2019 Feb 6;67(5):1418-1428. doi: 10.1021/acs.jafc.8b06207. Epub 2019 Jan 28.
Proanthocyanidins (PAs, also called condensed tannins), are an important class of secondary metabolites and exist widely in plants. Tea ( Camellia sinensis) is rich in PAs and their precursors, (-)-epicatechin (EC) and (+)-catechin (C). The biosynthesis of PAs is constantly regulated by many different MBW complexes, consisting of MYB transcription factors (TFs), basic-helix-loop-helix (bHLH) TFs, and WD-repeat (WDR) proteins. These regulatory factors can be environmentally affected, such as by biotic and abiotic stresses. In this study, we revalidated the effect of sucrose treatment on tea branches, and a sucrose-induced MYB (SIMYB) TF was screened and studied. Phylogenetic analysis indicted that this SIMYB TF belonged to MYB subgroup 5, named CsMYB5b. Heterologous expression of CsMYB5b in tobacco strongly induced PA accumulation, through up-regulating the key target genes LAR or ANRs. In addition, CsMYB5b restored PA production in the seed coat of A. thaliana tt2 mutant and rescued its phenotype. Yeast two-hybrid assay demonstrated CsMYB5b can interact directly with CsTT8 (an AtTT8 ortholog) and CsWD40 protein. Linking to the expression profiling of CsMYB5b and the PA accumulation pattern in tea plants suggest that the CsMYB5b acts as an important switch for the synthesis of monomeric catechins and PAs. Therefore, these data provide insight into the regulatory mechanisms controlling the biosynthesis of PAs.
原花青素(PAs,也称为缩合单宁)是一类重要的次生代谢产物,广泛存在于植物中。茶(Camellia sinensis)富含 PAs 及其前体物质,(-)-表儿茶素(EC)和(+)-儿茶素(C)。PAs 的生物合成不断受到许多不同的 MBW 复合物的调节,这些复合物由 MYB 转录因子(TFs)、碱性螺旋-环-螺旋(bHLH)TFs 和 WD-重复(WDR)蛋白组成。这些调节因子可能会受到环境的影响,例如生物和非生物胁迫。在本研究中,我们重新验证了蔗糖处理对茶树枝条的影响,并筛选和研究了一种蔗糖诱导的 MYB(SIMYB)TF。系统发育分析表明,这种 SIMYB TF 属于 MYB 亚组 5,命名为 CsMYB5b。CsMYB5b 在烟草中的异源表达强烈诱导了 PA 的积累,通过上调关键靶基因 LAR 或 ANRs。此外,CsMYB5b 恢复了拟南芥 tt2 突变体种皮中 PA 的产生,并挽救了其表型。酵母双杂交实验表明 CsMYB5b 可以与 CsTT8(AtTT8 的同源物)和 CsWD40 蛋白直接相互作用。与 CsMYB5b 的表达谱和茶树中 PA 积累模式的关联表明,CsMYB5b 作为单体儿茶素和 PAs 合成的重要开关。因此,这些数据为调控 PAs 生物合成的机制提供了新的见解。