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为研究在水、旱稻差异表达的抗旱候选基因、解释旱稻抗旱的分子机理,本研究选取已获得的水、旱稻干旱胁迫处理基因表达谱中在旱稻中高表达的候选基因,对其编码区和启动子区进行测序分析,以期比较可能的抗旱相关基因在水、旱稻基因组水平上的异同。根据水、旱稻差异表达结果,筛选出8个旱稻高表达基因。通过测序分析发现7个基因在水、旱稻间没有差异,只有一个候选基因OsMIP在典型水稻(日本晴、越富)和典型旱稻(IRAT109、毫格劳)启动子区之间存在18个SNP和1个Deletion的序列差异,说明在干旱胁迫下,旱稻高表达基因中只有少数存在序列上的差异。为进一步验证OsMIP的启动子区在水、旱稻间的序列差异,利用国内、外86份水、旱稻材料对该基因的启动子区进行测序分析,所得到的所有启动子序列可分为4种单倍型H1、H2、H3和H4,在单倍型H2中获得与抗旱相关的8个SNP和1个Indel,推测该基因的启动子区域的一个或多个序列差异可能是引起水、旱稻OsMIP基因差异表达的原因。
In order to study the drought resistance genes differentially expressed in water and upland rice and to explain the molecular mechanism of drought resistance in upland rice, we selected the candidate genes that were highly expressed in upland rice during drought stress treatment. Sub-region sequencing analysis, in order to compare the possible drought-related genes in water, upland rice genomics level similarities and differences. According to the differential expression results of water and upland rice, eight high-expression genes of upland rice were screened out. There was no difference between the seven genes in water and upland rice by sequencing analysis. Only one candidate gene OsMIP had 18 SNPs and 1 between the typical rice (Nipponbare and Yuefu) and typical upland rice (IRAT109, Sequence differences of Deletion indicated that there were only a few sequence differences between up-regulated genes in upland rice under drought stress. In order to further verify the sequence differences of promoter regions of OsMIP in water and upland rice, 86 promoter regions of OsMIP and Upland rice were sequenced and analyzed, and all the promoter sequences were classified into 4 types Haplotypes H1, H2, H3 and H4, eight SNPs and one Indel related to drought resistance were obtained in haplotype H2, suggesting that one or more sequence differences in the promoter region of the gene may be caused by water, The reason of OsMIP gene differential expression.