Expression and functional analysis of drought resistance related gene SsREMO-1a in Saccharum spontaneum L.
[Objective]The present paper aimed to obtain the complete sequence of S.sREMO-1a gene,conduct tissue-specific expression,qPCR expression analysis,sub-cellular localization study,and the phenotypic changes of Arabidopsis thaliana with SsREMO-1a gene transfer and the expression pattern of CAT-1 gene.[Method]The complete sequence of SsREMO-1a gene was obtained by PCR,purification,cloning and sequencing from Hainan sugarcane wild species(Sp-24).Drought stress was applied to the normal growth of the seedlings.Young leav-es,roots and stems at 0,1,2,3,4 and 5 days after the stress treatment were collected for tissue-specific expression analysis and qPCR a-nalysis.The tissue location and function of SsREMO-1a were analyzed by tobacco sub-cellular mapping and gene transformation techniques.pBI221-SsREMO-1a-GFP fusion expression vector was constructed and transformed into tobacco for cell localization analysis.The over-ex-pression vector pCAMBIA 1304-SsREMO-1a was constructed,and SsREMO-1a was introduced into A.thaliana by agrobacterium transforma-tion.The phenotypes of plants at 0,1,2,3,4 and 5 days after drought stress and the expression of CAT-1 gene were observed.[Result]The full length of SsREMO-1a gene was 738 bp,and the open code frame(ORF)was 564 bp,encoding 187 amino acids.Tissue-specific expression showed that SsREMO-1a was expressed in roots,stems and leaves.qPCR results showed that the gene was induced by drought stress and showed significantly up-regulated expression.The results of sub-cellular localization showed that SsREMO-1a protein was localized on the cell membrane.A total of 11 transgenic A.thaliana lines were obtained,and the drought resistance identification showed that the transgenic plants could improve the drought resistance of the plants.The expression of CAT-1 gene in transgenic plants was significantly in-creased.[Conclusion]SsREMO-1a can improve the drought resistance of plants and regulate CAT-1 gene to some extent.
万方数据
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