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在 和 中 CoHMGS 的表达分析及其编码酶的亚细胞定位。

Expression Analysis of CoHMGS in and Subcellular Localization of the Enzyme It Encodes.

机构信息

Agricultural College, Henan University of Science and Technology, Luoyang, China.

The Luoyang Engineering Research Center of Breeding and Utilization of Dao-di Herbs, Luoyang, China.

出版信息

DNA Cell Biol. 2023 Feb;42(2):91-96. doi: 10.1089/dna.2022.0527. Epub 2023 Feb 2.

Abstract

is a perennial deciduous tree or shrub. Its mature fruits are extracted and used in Traditional Chinese Medicine, called Shanzhuyu. The characteristic active components of include loganin and morroniside, which belong to iridoid glycosides. 3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is a key enzyme in the cytoplasmic mevalonate pathway providing the precursor molecules isopentenyl pyrophosphate and dimethylallyl pyrophosphate for isoprenoid biosynthesis such as sterols, triterpenes, and their derivatives such as iridoid glycosides. Different concentrations of methyl jasmonate (MeJA) and ethephon (ETH) solutions were sprayed on seedlings, and the effect of hormones on gene expression was detected by real-time fluorescence quantitative PCR. The quantitative real-time PCR results showed that 750 mg/L ETH treatment had the most significant induction effect on gene expression. The HPLC analysis of extracts revealed that the treatment could also significantly increase the content of morroniside and loganin in the leaves of . By use of a CoHMGS-green fluorescent protein (GFP) fusion construct for heterologous expression in tobacco, laser scanning confocal microscopy revealed a cytoplasmic localization. This preliminary study of the gene could prepare the ground for more precisely elucidating the synthesis of secondary metabolite in .

摘要

是一种多年生落叶乔木或灌木。其成熟果实经提取后用于中药,称为山茱萸。 的特征性活性成分包括马钱苷和莫诺苷,它们属于裂环环烯醚萜苷。3-羟-3-甲基戊二酰基辅酶 A 合酶(HMGS)是细胞质甲羟戊酸途径中的一种关键酶,为异戊烯基焦磷酸和二甲基烯丙基焦磷酸等前体分子提供萜类化合物、三萜及其衍生物如裂环环烯醚萜苷的生物合成。不同浓度的茉莉酸甲酯(MeJA)和乙烯利(ETH)溶液被喷洒在 幼苗上,通过实时荧光定量 PCR 检测激素对 基因表达的影响。定量实时 PCR 结果表明,750mg/L ETH 处理对 基因表达的诱导作用最显著。提取物的 HPLC 分析表明,该处理还可以显著增加 叶片中叶苷和马钱苷的含量。通过使用 CoHMGS-绿色荧光蛋白(GFP)融合构建体在烟草中进行异源表达,激光共聚焦显微镜显示其定位于细胞质。对 基因的初步研究为更精确地阐明 中次生代谢物的合成奠定了基础。

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