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VaCPK20基因的过表达显著增加了山葡萄细胞培养物中白藜芦醇的含量以及芪合酶基因的表达。

VaCPK20 gene overexpression significantly increased resveratrol content and expression of stilbene synthase genes in cell cultures of Vitis amurensis Rupr.

作者信息

Aleynova-Shumakova O A, Dubrovina A S, Manyakhin A Y, Karetin Y A, Kiselev K V

机构信息

Laboratory of Biotechnology, Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, Vladivostok, 690022, Russia.

出版信息

Appl Microbiol Biotechnol. 2014 Jun;98(12):5541-9. doi: 10.1007/s00253-014-5625-7. Epub 2014 Mar 4.

Abstract

Resveratrol, a naturally occurring plant phenol, has been reported to exhibit a wide range of valuable biological and pharmacological properties. In the present investigation, we show that transformation of a Vitis amurensis Rupr. cell suspension with the gene VaCPK20 for a calcium-dependent protein kinase (CDPK) under the control of double CaMV 35S promoter increased resveratrol production in five independently transformed cell lines in 9-68 times compared with control cells. The VaCPK20-transformed calli were capable of producing 0.04-0.42 % dry wt. of resveratrol, while the control calli produced up to 0.008 % dry wt. of resveratrol Also, we characterized expression of stilbene synthase (STS) genes in the five VaCPK20-transgenic cell lines of V. amurensis. In all VaCPK20-transgenic cell lines, expression of VaSTS7 increased; while expression of VaSTS1 decreased. We suggest that transformation of V. amurensis calli with the VaCPK20 gene induced resveratrol accumulation via enhancement of expression of the VaSTS7 gene involved in resveratrol biosynthesis. The obtained data first demonstrate that overexpression of a CDPK gene resulted in increased accumulation of a stilbenoid phytoalexine in transgenic plant cells. We propose that the VaCPK20 gene could play an important role in the regulation of resveratrol biosynthesis in grape cells.

摘要

白藜芦醇是一种天然存在的植物酚类物质,据报道具有广泛的重要生物学和药理学特性。在本研究中,我们发现,在双CaMV 35S启动子的控制下,用钙依赖蛋白激酶(CDPK)的VaCPK20基因转化山葡萄(Vitis amurensis Rupr.)细胞悬浮液,与对照细胞相比,五个独立转化的细胞系中白藜芦醇的产量提高了9至68倍。经VaCPK20转化的愈伤组织能够产生0.04 - 0.42%干重的白藜芦醇,而对照愈伤组织产生的白藜芦醇最高为0.008%干重。此外,我们还对山葡萄五个VaCPK20转基因细胞系中芪合酶(STS)基因的表达进行了表征。在所有VaCPK20转基因细胞系中,VaSTS7的表达增加;而VaSTS1的表达下降。我们认为,用VaCPK20基因转化山葡萄愈伤组织通过增强参与白藜芦醇生物合成的VaSTS7基因的表达诱导了白藜芦醇的积累。所获得的数据首次证明,CDPK基因的过表达导致转基因植物细胞中芪类植保素的积累增加。我们提出,VaCPK20基因可能在葡萄细胞中白藜芦醇生物合成的调控中发挥重要作用。

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