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在模式植物拟南芥和烟草中过表达的茶叶中存在的四种类黄酮糖基转移酶,用于功能鉴定。

Four flavonoid glycosyltransferases present in tea overexpressed in model plants Arabidopsis thaliana and Nicotiana tabacum for functional identification.

机构信息

State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, Anhui, China.

School of Life Science, Anhui Agricultural University, Hefei, Anhui, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Nov 15;1100-1101:148-157. doi: 10.1016/j.jchromb.2018.09.033. Epub 2018 Oct 7.

Abstract

Tea possesses a distinctive flavor profile and can have health benefits owing to the high levels of flavonoids in its leaves. However, the mechanism of the flavonoid glycosylation hasn't been well studied in tea plants, especially glycosylation at the 7-OH site has rarely been reported. In this study, four UGT genes CsUGT73A20, CsUGT75L12, CsUGT78A14 and CsUGT78A15 were isolated from tea leaves and overexpressed in the model plants Arabidopsis thaliana and Nicotiana tabacum for the functional identification of genes in vivo. In order to characterize the CsUGT functions in model plants, flavonoids in seeds of Arabidopsis and the flowers of tobacco were identified first. In CsUGT73A20-overexpressing Arabidopsis and tobacco, the level of certain flavonol glycosides involved in glycosylation reactions at the 3-OH and 7-OH sites increased considerably, but the level of flavan-3-ols decreased. In CsUGT75L12 transgenic Arabidopsis, the level of flavonol glycosides exhibiting glucosyltransferase activity at the 7-OH position increased markedly, but the concentrations of quercetin and kaempferol and flavan-3-ols decreased. In both transgenic Arabidopsis and tobacco, CsUGT78A14 promoted the synthesis of more flavonol glucosides with UDP-glucose as a sugar donor at the 3-OH glycosylation site. In CsUGT78A15 transgenic plants, flavonol galactosides at the 3-OH glycosylation site with UDP-galactose as a sugar donor were increased. In the tea plant, the corresponding flavonoid glycosides such as kaempferol‑3‑O‑β‑d‑glucosides, kaempferol‑3‑O‑β‑d‑galactosides, kaempferol‑7‑O‑β‑d‑glucoside, and luteolin‑7‑O‑β‑d‑glucoside were identified. And it could be possible that they were products of CsUGT78A14, CsUGT78A15, CsUGT73A20 and CsUGT75L12, respectively.

摘要

茶具有独特的风味,并因其叶片中含有高水平的类黄酮而具有健康益处。然而,茶树中类黄酮的糖基化机制尚未得到很好的研究,特别是在 7-OH 位的糖基化很少有报道。在这项研究中,从茶叶中分离出四个 UGT 基因 CsUGT73A20、CsUGT75L12、CsUGT78A14 和 CsUGT78A15,并在模式植物拟南芥和烟草中过表达,以在体内鉴定基因的功能。为了在模式植物中表征 CsUGT 的功能,首先鉴定了拟南芥种子和烟草花中的类黄酮。在 CsUGT73A20 过表达的拟南芥和烟草中,涉及 3-OH 和 7-OH 位糖基化反应的某些黄酮醇糖苷的水平显著增加,但黄烷-3-醇的水平降低。在 CsUGT75L12 转基因拟南芥中,7-OH 位具有葡萄糖基转移酶活性的黄酮醇糖苷水平显著增加,但槲皮素、山柰酚和黄烷-3-醇的浓度降低。在转基因拟南芥和烟草中,CsUGT78A14 促进了以 UDP-葡萄糖为糖供体的 3-OH 糖基化位点上更多黄酮醇糖苷的合成。在 CsUGT78A15 转基因植物中,以 UDP-半乳糖为糖供体的 3-OH 糖基化位点上的黄酮醇半乳糖苷增加。在茶树中,鉴定到相应的类黄酮糖苷,如山柰酚-3-O-β-D-葡萄糖苷、山柰酚-3-O-β-D-半乳糖苷、山柰酚-7-O-β-D-葡萄糖苷和木樨草素-7-O-β-D-葡萄糖苷。它们可能分别是 CsUGT78A14、CsUGT78A15、CsUGT73A20 和 CsUGT75L12 的产物。

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