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作者	单位
杨苑，肖磊，汪宇迪，谢利萍，黄媛媛，郭蔼光，徐虹	（1.西北农林科技大学生命科学学院，陕西杨凌 712100；2.旱区作物逆境生物学国家重点实验室，陕西杨凌 712100）

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中文摘要:

为建立小麦叶绿体基因启动子甲基化的检测体系，检测低温胁迫下小麦叶绿体编码基因甲基化的变化规律并分析叶绿体编码基因表达的调控模式，以低温敏感型小麦返白系及其对照矮变1号为材料，选取4个在低温条件下两个材料间表达有差异的叶绿体基因petN、trnC、petD和rrn16S，通过亚硫酸盐测序法(BSP-seq) 分别测定低温处理后这些基因(TaMET1, TaDRM和TaCMT)启动子的甲基化率，并用qPCR的方法对这4个基因以及预测定位于叶绿体的三个DNA甲基转移酶基因进行表达量检测。结果显示，叶绿体基因组总的甲基化水平在低温条件下持续增加，基因间启动子甲基化水平变化并不完全一致，基因型之间也略有差异。4个基因的表达量与甲基化水平的相关性不同，petN的表达对甲基化比较敏感，低温诱导甲基化率增加，基因的表达量则下调，但其余3个基因的表达与甲基化率的变化无直接相关性。返白系中三个DNA甲基转移酶基因的表达在低温条件下都呈上调趋势，但矮变1号中的 TaMET1与TaCMT基因表达下调。结果为深入研究低温下小麦叶绿体基因甲基化的变化规律及其对叶绿体基因的调控机理奠定了基础。

英文摘要:

In order to explore the variation of promoter methylation in chloroplast coding genes, and understand the regulatory mode of chloroplast gene expression under low temperature stress, we used the low temperature-sensitive Albinism line and Aibian 1 as materials.Four chloroplast genes(petN, trnC, petD and rrn16S) were selected, which were differentially expressed in Albinism line and Aibian 1 under cold stress.We measured the promoter methylation by the Bisulfite sequencing PCR(BSP-seq) at low temperature, and detected the expression of these chloroplast genes and three DNA methylation transferase genes(TaMET1, TaDRM and TaCMT) by qPCR, which are predicted to be located in the chloroplast.The results showed that the total methylation level of chloroplast genes continued to increase at low temperature, but the changes of methylation level among the promoters are not completely consistent, and there is also a slight difference between the two genotypes.The differences of the expression of four chloroplast genes are different from the changes of methylation.The expression of petN is more sensitive to methylation.Low temperature induced methylation rate increase and gene expression decrease.However, the expression of the other three genes have no direct correlation with changes of methylation rate.The expression of three DNA methyltransferase genes in the Albinism line is up-regulated in cold, but the expression of TaMET1 and TaCMT is down-regulated in Aibian 1.These results laid the foundation for further study of the changes of chloroplast gene methylation in wheat at low temperature and the methylation regulation mechanism of chloroplast genes.

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