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转 rolC 基因葡萄细胞培养物中白藜芦醇含量和苯丙氨酸解氨酶及芪合酶基因的表达

Resveratrol content and expression of phenylalanine ammonia-lyase and stilbene synthase genes in rolC transgenic cell cultures of Vitis amurensis.

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. 2010 Oct;88(3):727-36. doi: 10.1007/s00253-010-2792-z. Epub 2010 Aug 4.

Abstract

Resveratrol, a naturally occurring polyphenol, has been reported to exhibit a wide range of valuable biological and pharmacological properties. In the present investigation, we show that transformation of Vitis amurensis Rupr. with the oncogene rolC of Agrobacterium rhizogenes increased resveratrol production in the two transformed callus cultures 3.7 and 11.9 times. The rolC-transformed calli were capable of producing 0.099% and 0.144% dry weight of resveratrol. We characterized phenylalanine ammonia-lyase (PAL) and stilbene synthase (STS) gene expression in the two rolC transgenic callus cultures of V. amurensis. In the rolC transgenic culture with higher resveratrol content, expression of VaPAL3, VaSTS3, VaSTS4, VaSTS5, VaSTS6, VaSTS8, VaSTS9, and VaSTS10 was increased; while in the rolC culture with lower resveratrol content, expression of VaPAL3 and VaSTS9 was increased. We suggest that transformation of V. amurensis calli with the rolС gene induced resveratrol accumulation via selective enhancement of expression of individual PAL and STS genes involved in resveratrol biosynthesis. We compared the data on PAL and STS gene expression in rolC transgenic calli with the previously obtained results for rolB transgenic calli of V. amurensis. We propose that the transformation of V. amurensis with the rolC and rolB genes of A. rhizogenes increased resveratrol accumulation through different regulatory pathways.

摘要

白藜芦醇是一种天然存在的多酚,已被报道具有广泛的有价值的生物学和药理学特性。在本研究中,我们表明,根癌农杆菌的 rolC 基因转化山葡萄(Vitis amurensis Rupr.)增加了两种转化愈伤组织培养物中白藜芦醇的产量 3.7 和 11.9 倍。rolC 转化的愈伤组织能够产生 0.099%和 0.144%的白藜芦醇干重。我们在山葡萄的两种 rolC 转基因愈伤组织培养物中表征了苯丙氨酸解氨酶(PAL)和芪合酶(STS)基因的表达。在 rolC 转基因培养物中,具有较高白藜芦醇含量的 VaPAL3、VaSTS3、VaSTS4、VaSTS5、VaSTS6、VaSTS8、VaSTS9 和 VaSTS10 的表达增加;而在 rolC 培养物中,VaPAL3 和 VaSTS9 的表达增加。我们认为,山葡萄愈伤组织的 rolC 基因转化通过选择性增强参与白藜芦醇生物合成的单个 PAL 和 STS 基因的表达诱导了白藜芦醇的积累。我们将 rolC 转基因愈伤组织中 PAL 和 STS 基因表达的数据与之前获得的山葡萄 rolB 转基因愈伤组织的结果进行了比较。我们提出,根癌农杆菌的 rolC 和 rolB 基因转化山葡萄通过不同的调控途径增加了白藜芦醇的积累。

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