Functional Analysis of Two Flavanone-3-Hydroxylase Genes from Camellia sinensis: A Critical Role in Flavonoid Accumulation.
Functional Analysis of Two Flavanone-3-Hydroxylase Genes from Camellia sinensis: A Critical Role in Flavonoid Accumulation.
复制标题
茶花中两个黄烷酮-3-羟化酶基因的功能分析：在类黄酮积累中的关键作用
作者：
Han Y;Huang K;Liu Y;Jiao T;Ma G;Qian Y;Wang P;Dai X;Gao L;Xia T
黄酮类化合物是茶树的主要次级代谢产物。黄烷酮-3-羟化酶（F3H）是植物中黄酮类化合物生物合成的关键酶。然而，它在中华茶树黄酮代谢中的作用还没有得到很好的研究。本研究克隆了两个来自中华皂苷的F3H，分别命名为CsF3Ha和CsF3Hb，其中CsF3Ha含有1107个碱基，编码368个氨基酸；CsF3Hb含有1071个碱基，编码357个氨基酸。酶活性分析表明，两种重组 CsF3H 酶在大肠杆菌中都能将柚皮苷和二碘柚醇分别转化为二氢山奈酚（DHK）和二氢槲皮素（DHQ）。表达谱显示，CsF3Ha和CsF3Hb在茶树嫩叶中高表达。在不同的非生物胁迫下，紫外线（UV）辐射、蔗糖和脱落酸（ABA）对这两种CsF3Hs有显著的诱导作用。在 CsF3Hs 转基因拟南芥的种子中，大多数黄酮醇苷和低聚原花青素的浓度显著增加，而单儿茶素衍生物的含量则下降。本研究揭示了CsF3Hs在茶树类黄酮生物合成中的关键作用。
Flavonoids are major secondary metabolites in Camellia sinensis. Flavanone-3-hydroxylase (F3H) is a key enzyme in flavonoid biosynthesis in plants. However, its role in the flavonoid metabolism in C. sinensis has not been well studied. In this study, we cloned two F3Hs from C. sinensis, named CsF3Ha and CsF3Hb, where CsF3Ha containing 1107 bases encoded 368 amino acids, and CsF3Hb containing 1071 bases encoded 357 amino acids. Enzymatic activity analysis showed both recombinant CsF3H enzymes in Escherichia coli could convert naringenin and eriodictyol into dihydrokaempferol (DHK) and dihydroquercetin (DHQ), respectively. The expression profiles showed that CsF3Ha and CsF3Hb were highly expressed in the tender leaves of tea plants. Under different abiotic stresses, the two CsF3Hs were induced remarkably by ultraviolet (UV) radiation, sucrose, and abscisic acid (ABA). In the seeds of CsF3Hs transgenic Arabidopsis thaliana, the concentration of most flavonol glycosides and oligomeric proanthocyanidins increased significantly, while the content of monocatechin derivatives decreased. The present study revealed that CsF3Hs played critical roles in flavonoid biosynthesis in tea plants.
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