DNA甲基化修饰在马铃薯响应低温胁迫中的调控作用

马铃薯生育周期较长，低温危害严重影响着其产量和质量。DNA 甲基化修饰在植物逆境胁迫中起重要作用，其通过对DNA序列的甲基化或去甲基化来调控基因表达进而响应逆境胁迫。但DNA甲基化修饰如何调控马铃薯响应低温胁迫的分子机制尚不清楚。本项目以国内马铃薯主栽品种‘大西洋’为研究对象，利用甲基化敏感扩增多态性（MSAP）技术分析了低温处理前后基因甲基化状态的变化，比较分析后得到12条可能参与马铃薯响应低温胁迫的差异片段，在此基础上，拟利用甲基化特异性PCR技术，确定参与响应马铃薯低温胁迫的受甲基化调控的基因，通过VIGS和RNAi转基因技术在烟草和马铃薯中确定基因功能，并利用real-time PCR和Confocal确定基因表达模式及其在细胞中作用场所及位点，确定参与调控马铃薯响应低温胁迫的受DNA甲基化调控的基因及其作用机制，为马铃薯耐受低温育种与生产提供基础，保证我国马铃薯高产丰收。

The lower temperature lays negative impacts on potato (Solanum tuberosum L.) production for it needs a long growth period to grow and mature. DNA methylation plays an important roles in regulation of plant adaptation to enviromental stresses. It regulates gene expression by methylation or de methylation of DNA sequence and then responds to stress. However, the molecular mechanism of DNA methylation modification is not clear. In this study, 12 different fragments may be involved in the response to low temperature stress of potato cv. Atlantic were isolated by methylation sensitive amplified polymorphism (MSAP). According to the above data, the further experiments will be designed to determinate the methylation regulated genes involved in response to low temperature stress in potato by methylation-specific PCR (MS-PCR). Virus induced gene silencing (VIGS) and RNAi will be used in this research to explore the roles and functions of these methylation regulated DNA bands play in response to lower temperature in potato. Real-time PCR and Confocal will be used to determinate expression patterns and action site of genes in cells. The aims of this research are to identify genes modulated by DNA methylation and to explore the roles of these genes playing under lower temperature stress in potato, and eventually to benefit the development of potato molecular genetic breeding and production.