水稻品种耐冷适应性与ICE1基因甲基化变异关系的研究

发布时间：2018-03-17 14:09

本文选题：水稻　切入点：ICE基因　出处：《农业生物技术学报》2017年09期 　论文类型：期刊论文

【摘要】：水稻(Oryza sativa)起源于热带,随着人们对粮食需求的增长其种植区不断向北扩展,这就要求选择耐冷能力强的水稻品种。但是目前,对水稻耐冷性的分化及其机制的研究尚少见报道。本研究以辽宁、江苏和广东3个不同气候地区水稻为实验材料,通过冷害症状及叶绿素荧光指数比较水稻耐冷性差异,采用DNA甲基化亚硫酸盐测序法(bisulfite sequencing PCR,BSP),研究CBF(C-repeat binding factor)冷响应转录通路的ICE1(inducer of CBF expression 1)基因区域与启动子区域甲基化水平影响不同品种水稻冷害差异的分子机制。结果表明ICE1基因区域甲基化水平无差异,但是启动子区域甲基化有显著差异(P0.05)。辽宁丹粳17(LN004)的ICE1基因启动子区域2个胞嘧啶被甲基化,耐冷指数最高;江苏的南粳5055(JS013)的ICE1基因启动子区域7个胞嘧啶被甲基化,耐冷指数居中;广东的粤新占2号(GD008)的ICE1基因启动子区域11个胞嘧啶被甲基化,耐冷指数为最低。定量PCR检测结果显示,ICE1和其下游调控的CBF1(C-repeat DRE binding factor 1)、CBF3(C-repeat DRE binding factor 3)的基因表达量与耐冷性显著正相关(P0.05),但是与ICE1启动子区域甲基化水平显著负相关(P0.05)。这表明辽宁(LN004)、江苏(JS013)和广东(GD008)水稻的耐冷性与地区气候相适应,且耐冷性受ICE1基因甲基化水平决定的CBF冷响应转录通路调节。本研究结果为进一步解释水稻耐冷的分子机制提供了理论依据。
[Abstract]:Rice Oryza sativa originated in the tropics, and its growing area is expanding northward as the demand for food increases, which requires the selection of rice varieties with strong cold tolerance. Studies on the differentiation and mechanism of cold tolerance in rice were rarely reported. In this study, rice chilling tolerance was compared by chilling injury symptoms and chlorophyll fluorescence index in three different climatic regions of Liaoning, Jiangsu and Guangdong. DNA methylated sulfite sequencing was used to study the molecular mechanism of CBF(C-repeat binding factor cold response ICE1(inducer of CBF expression 1) gene region and promoter region methylation level affecting chilling injury difference in different rice varieties. The results show that the molecular mechanism of the methylation level of the ICE1(inducer of CBF expression 1 gene region in the cold response transcription pathway of CBF(C-repeat binding factor is studied. There was no difference in methylation level of ICE1 gene region. However, there was a significant difference in promoter region methylation (P0.05N). Two cytosines were methylated in the promoter region of ICE1 gene in Danjing, Liaoning Province, and 7 cytosines were methylated in the promoter region of ICE1 gene in Nanjing5055JS 013 (Jiangsu Province). The cold tolerance index was in the middle. 11 cytosines were methylated in the promoter region of ICE1 gene in Yuexin No. 2, Guangdong, China. The results of quantitative PCR analysis showed that the gene expression of ICE1 and its downstream CBF1(C-repeat DRE binding factor (CBF3C-repeat DRE binding factor 3) was positively correlated with cold tolerance (P0.05A), but negatively correlated with the methylation level of ICE1 promoter region (P0.05). This indicates that the cold tolerance of rice in Liaoning (LN004), Jiangsu (JS013) and Guangdong (GD008) adapts to the regional climate. The cold tolerance is regulated by the CBF cold response transcriptional pathway, which is determined by the methylation level of the ICE1 gene. This study provides a theoretical basis for further explanation of the molecular mechanism of cold tolerance in rice (Oryza sativa L.).
【作者单位】：南京农业大学生命科学学院;
【基金】：国家自然科学基金(No.31572066) 转基因生物研究与商业化重大专项(No.2016ZX08011-001) 教育部高等学校博士点基金优先发展领域课题(No.20130097130006)
【分类号】：S511

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