水稻逆境响应基因OsMsr11的克隆与分析

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。

Cloning and Analysis of Stress Responsive Gene OsMsr11 in Rice

To discover new stress tolerance genes,OsMsr11(Oryza sativa L.multiple stress responsive gene 11) was chosen to be cloned and analyzed,based on the analysis of global gene expression profiles of rice Pei’ai 64S under heat,drought and cold stresses by using GeneChip Rice Genome Array(Affymetrix).OsMsr11 was a cold-,drought-and heat-inducible gene,and the expression level was highly up-regulated by cold stress at the heading stage.The cDNA of OsMsr11 was cloned through PCR amplification.Sequence analysis showed that the cDNA consists of an open reading frame of 234 bp,encoding a protein of 77 amino acid residues.The encoded protein contains a signal peptide and may be secreted into the periplasmic space.Known-function domain was not observed in the protein sequence.The OsMsr11 gene lacks intron.Analysis of the putative promoter region identified matches to cis-elements related to stress responses.To analyze its function,the OsMSR11 gene was transferred into rice 9311 by Agrobacterium-mediated method.The obtained data showed that over-expression of OsMsr11 in rice enhanced the tolerance of transgenic plants to salt and reduced their sensitivity to ABA.