Functional Analysis Of Rice Stress Assicoliate Protein OsiSAP1 Responsed To Salt-alkali Stress

OsiSAP1(Rice stress associated proteins)is a member of zinc finger protein transcription factor family,and contains A20/AN1 zinc finger domain.In plant,OsiSAP1 was the firstly identified gene which was related to stress.The OsiSAP1 was played role in the regulation mechanisms which were induced by salt/alkality,cold,desiccation,submergence,drought,heavy metals and mechanical wounding in rice.Also,it was involoved in the regulation of pathogenic mechanism of bacteria,hormonic and glycometabolism.In present study,the OsiSAP1 gene was cloned by RT-PCR technique from longjing11.The open reading frame(ORF)of OsiSAP1 is 495 bp,encoded 164 amino acids,and contains an A20/AN1 zinc finger domain.Phylogenetic analysis showed it had the highest homology with OsiSAP1 and Aegilops tauschii AgeSAP1 gene.Real-time PCR results showed that OsiSAP1 was expressed in root,leaf,panicle,shell,flower,stem,panicle stem and flag leaf.The OsiSAP1 expression levels in root and leaves are in an increasing trend during the salt(NaCl)and alkali(NaHCO3)stress condition.The binary vector(pBS-OsiSAP1-GFP)was imported into onion epidermal cells by gene gun,and the fluorescence microscope result showed that the OsiSAP1::GFP fusion protein was located in nucleus of onion epidemal cells.The overexpression OsiSAP1 Arabidopsis and Rice have been obtained.Treated with NaCl and NaHCO3 medium,the results revealed that the growing situation and fresh weight in overexpression OsiSAP1 Arabidopsis were better than that of wild-type at germination and late germination.At late germination,the MDA(malondialdehyde)content of overexpression OsiSAP1 Arabidopsis is less than that of wild-type,and the growing situation of overexpression OsiSAP1 Arabidopsis is better than that of wild-type.All of those indicated that OsiSAP1 was involved in the regultation of salinealkali resistance and improving the antioxidant capacity in plants.In the experiment of overexpression OsiSAP1 rice,the results showed that growing situation of transgene rice was better than that of wild-type in the four saline-alkali treated groups,and the stem height,root length and resh weight of overexpression OsiSAP1 rice were better than that of wild-type.The results indicated that overexpression OsiSAP1 could improve salt and alkality stress tolerance in rice.Moreover,the overexpression OsiSAP-FLAG rice was obtained in present study,which will be contribute to feather research.The protein,pyruvate kinase,was screened by yeast twohybrid which was verified to interact with OsiSAP1..