温度胁迫下侧耳DNA甲基化动态变化及其生物学效应研究

项目名称： 温度胁迫下侧耳DNA甲基化动态变化及其生物学效应研究

项目编号： No.31201667

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 黄晨阳

作者单位： 中国农业科学院农业资源与农业区划研究所

项目金额： 20万元

中文摘要： 高温和低温是食用菌经常遭受的两种主要环境胁迫。DNA甲基化和去甲基化是生物体应对各种逆境的重要机制之一。本项目拟以糙皮侧耳、肺形侧耳、白灵侧耳为材料，设置不同温度、时间、次数、菌株等条件，采用甲基化敏感扩增多态性(MSAP)技术对温度胁迫后DNA甲基化水平变化进行研究，解析温度胁迫后DNA甲基化的群体一致性、稳定性、生物学效应和在后代中的遗传规律。采用切胶回收、序列测定和网络比对，对DNA甲基化的差异片段进行定性分析。采用Bio-COBRA(联合亚硫酸氢钠限制性内切酶分析法和Agilent 2100 Bioanalyzer结合)，对特定片段进行定量分析。通过研究食用菌中DNA甲基化/去甲基变化，探讨低温胁迫与菌种的遗传不稳定的内在联系，以及高温胁迫与子实体农艺性状等恶化的关系。本研究将为菌种保藏研究提供科学数据，为菌种质量监测、高效生产提供技术支撑。

中文关键词： 表观遗传；甲基化敏感扩增多态性；平菇；全基因组；

英文摘要： The growth of edible mushroom is susceptible to environmental stress, especially heat stress and cold stress. DNA methylation and demethylation is one of the important mechanisms in response to environmental stress for many organisms. Pleurotus ostreatus, Pleurotus pulmonarius and Pleurotus eryngii var. tuoliensis will be used as materials in this study. Different temperatures, time, times and strains will be investigated, Methylation-sensitive Amplification Polymorphism (MSAP) technology will be adopted to detect DNA methylation level under different conditions. By analysis, uniform, stability, biology effects and inheritance in offspring of Pleurotus DNA methylation will be figure out. Differential bands gained by gel extraction will be sequenced and qualitative analyzed by Blast. Quantitative analysis will be done by Bio-COBRA. The relation between cold stress and genetically instability would be discussed based on changes of DNA methylation and demethylation in edible mushroom as well as the relation between heat stress and deterioration of agronomic traits of edible mushrooms. This study is supposed to provide fundamental data for culture preservation of edible mushrooms, and it can give technological support for monitoring of spawn quality and high-yields cultivation.

英文关键词： Epigenetics；Methylation sensitive amplification polymorphism；Pleurotus ostreatus；whole genome；