参与山茱萸铱苷合成的关键酶基因 CoGPPS 的克隆和表达分析

DNA and cell biology Pub Date : 2024-03-01 Epub Date: 2024-02-13 DOI:10.1089/dna.2023.0335
Jiaxi Chen, Xinjie Tan, Guangyang Guo, Panpan Wang, Hongxiao Zhang, Shufang Lv, Huawei Xu, Dianyun Hou
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引用次数: 0

摘要

山茱萸鸢尾甙(CIGs),包括山茱萸素和山茱萸苷,是山茱萸的主要活性成分。作为 CIGs 生物合成过程中的关键酶之一,焦磷酸香叶酯合成酶(GPPS)催化焦磷酸香叶酯的形成,而焦磷酸香叶酯是 CIGs 的直接前体。本研究克隆并分析了 C. officinalis 的焦磷酸香叶酯合成酶(CoGPPS)序列。CoGPPS 基因的 cDNA 序列为 915 bp(GenBank 编号:OR725699)。系统进化分析表明,CoGPPS 与放线菌和山茶的 GPPS 序列亲缘关系密切,但与芍药和三尖杉的 GPPS 序列亲缘关系相对较远。实时定量 PCR 的结果显示了 CoGPPS 的时空表达模式,即 CoGPPS 在果实中特异性表达。亚细胞定位检测证明,CoGPPS特异性地存在于叶绿体中。通过高效液相色谱法检测了组织中的洛神花素和吗啉苷含量,发现果实中这两种化合物的含量均高于叶片。因此,本研究为进一步研究 CIG 的合成途径奠定了基础。
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Cloning and Expression Analysis of Key Enzyme Gene CoGPPS Involved in Iridoid Glycoside Synthesis in Cornus officinalis.

Cornus iridoid glycosides (CIGs), including loganin and morroniside, are the main active components of Cornus officinalis. As one of the key enzymes in the biosynthesis of CIGs, geranyl pyrophosphate synthase (GPPS) catalyzes the formation of geranyl pyrophosphate, which is the direct precursor of CIGs. In this study, the C. officinalis geranyl pyrophosphate synthase (CoGPPS) sequence was cloned from C. officinalis and analyzed. The cDNA sequence of the CoGPPS gene was 915 bp (GenBank No. OR725699). Phylogenetic analysis showed that CoGPPS was closely related to the GPPS sequence of Actinidia chinensis and Camellia sinensis, but relatively distantly related to Paeonia lactiflora and Tripterygium wilfordii. Results from the quantitative real-time PCR showed the spatiotemporal expression pattern of CoGPPS; that is, CoGPPS was specifically expressed in the fruits. Subcellular localization assay proved that CoGPPS was specifically found in chloroplasts. Loganin and morroniside contents in the tissues were detected by high-performance liquid chromatography, and both compounds were found to be at higher levels in the fruits than in leaves. Thus, this study laid the foundation for further studies on the synthetic pathway of CIGs.

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