Identification And Functional Analysis Under Low Temperature Stress Of Transcription Factor GATA16 In Rice

The transcription factors of GATA family in rice are involved in growth and development processes such as flowering,leaf formation,however the regulation roles of abiotic stress is not yet clear.In this study,we identified a unreported GATA transcription factor gene Os GATA16,and performed the research of function and molecular mechanism under low temperature stress.The results are as follows:1.Bioinformatics analysis results showed that Os GATA16 belongs to Os GATA family,subfamily-II,class-A,that protein has representative GATA-type zinc finger protein domain,nuclear localization signal(NLS),conserved LLM domain,and predict Os GATA16 has eleven cold-related phosphorylation sites.In GATA16 of different species,GATA-type zinc finger protein domains showed highly conserved in each species;in addition,Os GATA16 keeps the most homologous with Os GATA11.2.Phenotype analysis results showed that Os GATA16 overexpression transgenic lines(OEs)showed better in cold tolerance and difference of survival rate than wild type(WT)under cold treatment at 3-leaf stage in rice.Whereas under normal growth conditions,there no obvious difference between OEs and WT.3.Os GATA16 protein is a transcriptional suppressor as transcriptional regulator performs functions in nucleus.4.Expression pattern analysis results showed that Os GATA16 showed expressing in whole plant,and higher expression in young root,stem and panicle.For diverse treatments,Os GATA16 showed induced by cold,abscisic acid(ABA);suppressed by drought,cytokinin and jasmonic acid(JA),there no obvious change in salt treatment.5.Target protein Os VOZ1 was found by screening yeast c DNA library,and then Yeast Two-Hybrid and Bimolecular Fluorescence Complementation(BIFC)testified Os GATA16 interaction with Os VOZ1.6.Four tentative genes Os WRKY45-1,Os SRFP1,Os CYL4 and Os MYB30 that possible as target gene of Os GATA16 were found by q RT-PCR,and then Os GATA16 increases cold tolerance through binding and suppressing expression of Os WRKY45-1 in rice testified by Yeast One-Hybrid and Dual-Luciferase Reporter Assay System.