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过量表达 通过增强儿茶素竞争促进原花青素途径来抑制葡萄()细胞中的花青素生物合成。

Overexpression of Suppresses Anthocyanin Biosynthesis by Enhancing Catechin Competition Leading to Promotion of Proanthocyanidin Pathway in Spine Grape () Cells.

机构信息

Institute of Food Science and Technology, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China.

College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China.

出版信息

Int J Mol Sci. 2024 Nov 11;25(22):12087. doi: 10.3390/ijms252212087.

Abstract

Proanthocyanidins (PAs) are a class of polyphenolic compounds recognized for their potent antioxidant, anti-cancer, anti-inflammatory, and cardioprotective properties. However, the production of PAs from natural sources is often limited by high costs, resource wastage, and environmental damage. In this study, we investigated the overexpression of , along with phenotypic observation, metabolite determination, light quality treatment, and RT-qPCR analysis, in spine grape cells. The results demonstrated a significant increase in the contents of proanthocyanidins and flavonoids in p-overexpressing transgenic cell lines, while anthocyanin levels showed a decreasing trend. Furthermore, the treatment with white and blue light on the T5 cell line resulted in enhanced accumulation of proanthocyanidins, catechins, and flavonoids, whereas anthocyanins and epicatechins exhibited a declining pattern. Thus, short-wavelength light promoted the accumulation of metabolites, with the proanthocyanidin content in the T5 transformed cell line reaching 2512.0 μg/g (FW) during blue light incubation. RT-qPCR analysis revealed that the key genes involved in the biosynthesis of proanthocyanidin and anthocyanin were upregulated in the transgenic spine grape cell lines, with expression increasing by several hundredfold, far surpassing the expression levels of and . The overexpression markedly improved substrate competitiveness within the metabolic pathway, promoting catechin biosynthesis while inhibiting the production of epicatechins and anthocyanins. This finding provides compelling evidence that is a crucial gene for catechin biosynthesis. This research establishes both theoretical and practical foundations for the regulation and development of natural proanthocyanidins, addressing issues related to high costs, safety concerns, resource wastage, and environmental damage associated with their production.

摘要

原花青素(PAs)是一类具有强大抗氧化、抗癌、抗炎和心脏保护作用的多酚化合物。然而,从天然来源生产 PAs 常常受到高成本、资源浪费和环境破坏的限制。在这项研究中,我们调查了在刺葡萄细胞中过量表达 ,以及表型观察、代谢物测定、光质处理和 RT-qPCR 分析。结果表明,在过表达转基因细胞系中,原花青素和类黄酮的含量显著增加,而花青素水平呈下降趋势。此外,对白光和蓝光处理 T5 细胞系导致原花青素、儿茶素和类黄酮的积累增加,而花青素和表儿茶素则呈下降趋势。因此,短波长光促进了代谢物的积累,在蓝光孵育下,T5 转化细胞系中原花青素的含量达到 2512.0 μg/g(FW)。RT-qPCR 分析显示,参与原花青素和花青素生物合成的关键基因在转基因刺葡萄细胞系中上调, 表达增加了数百倍,远远超过 和 的表达水平。 的过表达显著提高了代谢途径中的底物竞争力,促进儿茶素的生物合成,同时抑制表儿茶素和花青素的产生。这一发现为 是儿茶素生物合成的关键基因提供了有力证据。这项研究为天然原花青素的调控和开发奠定了理论和实践基础,解决了与生产相关的高成本、安全性问题、资源浪费和环境破坏等问题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e492/11594265/8ec250513e6c/ijms-25-12087-g001.jpg

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