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为了克隆和研究逆境诱导型水稻启动子的功能,根据前期工作所获得的高温条件下水稻幼穗的表达谱数据,结合对GEO数据库中有关水稻逆境芯片数据的检索与分析,选取了3个对高盐和高温显著响应的基因(Os11g0453900,Os09g0526600,Os05g0381400)。基于这3个基因ATG上游1.5kb的启动子序列的生物信息学分析,克隆了这3个启动子中包含有多个ABA、高温、高盐等逆境响应元件的DNA片段,分别命名为Rab16Dp、OsHsfB2cp和PM19p,并通过农杆菌介导转化烟草叶盘进行了瞬时表达分析。结果表明:高温、高盐对Rab16Dp和OsHsfB2cp启动子有较明显的诱导效应,而对PM19p的诱导效果不明显。实验为这3个启动子功能的进一步研究奠定了基础,也将为逆境诱导型水稻启动子的挖掘和鉴定提供重要线索。
In order to clone and study the function of adversity-inducible rice promoters, three pairs of primers were selected according to the expression profiles of rice panicle at high temperature obtained in the previous work, and the retrieval and analysis of the data of rice adversity chips in GEO database High salt and high temperature significant response of genes (Os11g0453900, Os09g0526600, Os05g0381400). Based on the bioinformatics analysis of the 1.5 kb upstream ATG of these three genes, we cloned the DNA fragments of these three promoters which contained multiple ABA, high temperature and high salt stress response elements, named Rab16Dp, OsHsfB2cp and PM19p, and transient expression analysis of tobacco leaf disks mediated by Agrobacterium was carried out. The results showed that high temperature and high salt had obvious inducing effects on Rab16Dp and OsHsfB2cp promoters, but no obvious effect on PM19p. The experiment lays the foundation for the further study on the function of these three promoters and will also provide important clues for the discovery and identification of adversity-inducible rice promoters.