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为了挖掘新的耐逆基因,本文在应用Affymetrix水稻表达芯片分析超级稻亲本培矮64S在不同逆境(高温、干旱、低温)胁迫下、不同发育时期叶片和穗中全基因组表达模式的基础上,对其中一个多逆境响应基因OsMsr11(Oryza sativa L.multiple stressresponsive gene 11)进行了克隆与分析。OsMsr11是一个受低温、干旱、高温多逆境诱导的基因,在孕穗期低温胁迫下表达水平显著上调。通过PCR扩增获得了包含其完整开放阅读框的cDNA序列。序列分析表明,其ORF为234 bp,编码77个氨基酸,有信号肽序列,无典型的保守基因结构域,预测其为分泌通路信号肽,分泌到细胞周质中,功能未知;OsMsr11基因不含内含子,对其启动子区域进行分析,发现含有多种与逆境响应相关的顺式作用元件。为研究其功能,采用农杆菌侵染法转化超级稻父本9311,初步分析表明:在水稻中过量表达OsMsr11,增强了转基因水稻的耐盐性,降低了对外源ABA的敏感性。
In order to explore new gene of stress tolerance, we analyzed the whole genome expression patterns of leaves and ears at different developmental stages under different stress (high temperature, drought and low temperature) stress using Affymetrix rice expression chip, One of the multi-stress response genes, OsMsr11 (Oryza sativa L. mplex stress responsive 11) was cloned and analyzed. OsMsr11 is a gene induced by multiple stresses of low temperature, drought and high temperature. The expression of OsMsr11 was significantly up-regulated under low temperature stress at booting stage. The cDNA sequence containing its complete open reading frame was obtained by PCR amplification. Sequence analysis showed that the ORF was 234 bp and encoded a polypeptide of 77 amino acids with a signal peptide sequence and no typical conserved domain. It was predicted to be a secretory pathway signal peptide and secreted into the periplasm of cells with unknown function. OsMsr11 gene contained no Analysis of its promoter region revealed that it contains multiple cis-acting elements that are associated with adverse stress responses. In order to study its function, Agrobacterium tumefaciens was used to transform male parent 9311. Preliminary analysis showed that overexpression of OsMsr11 in rice enhanced the salt tolerance of transgenic rice and reduced its sensitivity to exogenous ABA.