Laboratory of Biotechnology, Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, 690022, Vladivostok, Russia,
Plant Cell Rep. 2015 Feb;34(2):311-20. doi: 10.1007/s00299-014-1708-2. Epub 2014 Nov 25.
Salicylic acid (SA) treatment selectively reduced the cytosine DNA methylation of stilbene synthase ( STS ) genes and stimulated resveratrol production in cell cultures of Vitis amurensis. The effect of salicylic acid (SA) on plant growth, flowering time, and fruit number is known to correlate with the level of DNA methylation, while the potential correlation between SA-induced changes in DNA methylation and biosynthesis of secondary metabolites has not been studied. Trans-resveratrol, a naturally occurring plant phenol, has been reported to exhibit a wide range of valuable biological and pharmacological properties. In this study, cell cultures of Vitis amurensis capable of producing t-resveratrol were used as a model system to study whether the SA-induced increase in t-resveratrol production is associated with changes in DNA methylation of stilbene synthase (STS) genes. T-resveratrol is synthesized via the phenylpropanoid pathway, in which STS genes are the key enzymes. Treatment of V. amurensis callus cultures with SA significantly increased t-resveratrol production and the expression of certain STS genes (e.g., VaSTS2 and VaSTS10). A marked decrease in the methylation of the VaSTS2 and VaSTS10 genes in response to SA was demonstrated using bisulfite sequencing, while no considerable changes were detected in the methylation of VaSTS1, a constitutively and highly expressed STS gene. The obtained results show that SA treatment selectively reduced cytosine methylation of VaSTS genes. The data suggest that selective DNA demethylation of particular STS genes could be necessary for the activation of t-resveratrol biosynthesis in response to SA. This finding provides an insight into the mechanism of SA action and biosynthesis of secondary metabolites in plant cells.
水杨酸(SA)处理选择性地降低了苯乙烯合酶(STS)基因的胞嘧啶 DNA 甲基化,并刺激了山葡萄细胞培养物中白藜芦醇的产生。已知水杨酸(SA)对植物生长、开花时间和果实数量的影响与 DNA 甲基化水平相关,而 SA 诱导的 DNA 甲基化变化与次生代谢物生物合成之间的潜在相关性尚未得到研究。反式白藜芦醇,一种天然存在的植物酚,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,使用能够产生反式白藜芦醇的山葡萄细胞培养物作为模型系统,研究 SA 诱导的反式白藜芦醇产量增加是否与苯乙烯合酶(STS)基因的 DNA 甲基化变化相关。反式白藜芦醇通过苯丙氨酸途径合成,其中 STS 基因是关键酶。SA 处理山葡萄愈伤组织培养物显著增加了反式白藜芦醇的产生和某些 STS 基因(如 VaSTS2 和 VaSTS10)的表达。亚硫酸氢盐测序表明,VaSTS2 和 VaSTS10 基因对 SA 的响应表现出明显的甲基化降低,而 VaSTS1 是一个组成型且高度表达的 STS 基因,其甲基化没有明显变化。所得结果表明,SA 处理选择性地降低了 VaSTS 基因的胞嘧啶甲基化。数据表明,特定 STS 基因的选择性 DNA 去甲基化可能是对 SA 响应激活反式白藜芦醇生物合成所必需的。这一发现为理解植物细胞中 SA 作用和次生代谢物生物合成的机制提供了新的见解。