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为深入了解植物的逆境反应机理和发现新的水稻耐逆基因,采用Affymetrix水稻表达芯片(含51279个转录本)分析了超级稻两优培九母本培矮64S不同生长发育时期、不同组织器官全基因组在低温、干旱、高温逆境胁迫下的表达水平,筛选出众多表达水平显著升高与降低的基因。OsMsr1(Oryza sativa L.multiple stresses responsive gene 1,Gen Bank登录号为EU284112)是其中一个受高温、低温与干旱诱导、在各生长发育时期与组织器官均显著上调的基因,用实时定量PCR方法对其表达水平进行了进一步的分析,所得结果与基因芯片结果基本吻合,因此,此基因为一多逆境响应基因。用RT-PCR方法扩增获得了包含其完整ORF(open reading frame)的cDNA克隆。根据其ORF序列进行预测,此基因编码一个包含89个氨基酸残基的小分子蛋白,分子量约为10kD,pI约为5。搜索有关数据库,在水稻、玉米、小麦与拟南芥中找到有高相似性的基因,但功能未知,也未发现相同与类似的已知功能的基因保守结构域。对其可能的启动子序列分析,发现5个与逆境反应有关的顺式作用元件。因此,我们认为该基因为一新的水稻耐逆候选基因,进一步的研究正在进行。
In order to further understand the mechanism of plant stress response and find out new rice stress tolerance genes, Affymetrix rice expression chips (containing 51279 transcripts) were used to analyze the growth and development of Pei’ai Pei’ai 64S at super hybrid rice Liangyoupeijiu. The whole genome of different tissues and organs Low temperature, drought, high temperature stress under stress conditions, screening a large number of expression levels were significantly increased and decreased genes. OsMsr1 (Oryza sativa L. multiplex responsive gene 1, GenBank accession number EU284112) is one of the genes induced by high temperature, low temperature and drought, and significantly upregulated in all tissues and organs at different developmental stages. Real-time quantitative PCR The expression level was further analyzed, and the results were basically consistent with the results of the gene chip, therefore, this gene is a multi-stress response gene. The cDNA clone containing its complete ORF (open reading frame) was amplified by RT-PCR. According to its ORF sequence prediction, this gene encodes a small molecule protein containing 89 amino acid residues with a molecular weight of about 10 kD and a pI of about 5. In search of databases, genes with high similarity in rice, maize, wheat and Arabidopsis were found but their functions were unknown and no homologous and similar gene-conserved domains with known functions were found. Analysis of its possible promoter sequences revealed five cis-acting elements involved in adversity reactions. Therefore, we consider this gene as a new rice resistance candidate gene, and further research is underway.