Study On The Interaction Between Rice OsEIL1and OsEBFs

Studies have found that two kinds of Arabidopsis EIN3-binding F-box (EBF)1and2both have interaction with ETHYLENE INSENSITIVE3(EIN3) transcription factor. EBF1and EBF2overexpression can result in plants is not sensitive to ethylene. Instead, the EBF1and EBF2mutation plant has demonstrated continuous ethylene signal response and EIN3protein accumulation. It's reported that the tomato S1-EBF1and S1-EBF2play a key role during organ development. Overexpression of S1-EBF1and S1-EBF2during the flowering to fruit maturity. Silencing the S1-EBF1and S1-EBF2can speed ethylene phenotype up, such as accelerated senescence accelerated tomato fruit ripening. The above structure that EBF1and EBF2involved in plant ethylene signal transduction pathway, possibly through interactions protein in the signal transduction process and play an important role in plant ethylene signal transduction and plant growth and development.OsEIL1, which showed highest similarity with EIN3. Chuanzao Mao et had found that Ectopic expression of OsEIL1in Arabidopsis can partially complement the ein3-1mutant. The transgenic rice plants with overexpression of OsEIL1exhibit short root, coiled primary root and slightly short shoot phenotype and elevated response to exogenous ethylene. These results indicate that OsEIL1is involved in ethylene signal transduction pathway and acts as a positive regulator of ethylene response in rice.According to the above results, we have enough reasons to suspect that OsEBF1and OsEBF2mediate the ethylene signal transduction pathway, may through get interact with rice EIN3-like (OsEIL1) protein mediate ubiquitination pathways affected the stability of OsEIL1.In this study, the interactions between rice ethylene signal control factor OsEIL1and OsEBF1,OsEIL1and OsEBF2were examined via yeast two-hybrid system and GST pull-down. The main research results are as follows:(1) Three yeast expression vetors of rice ethylene signal control factor pGADT7-OsEIL1and pGBKT7-OsEBF1,pGBKT7-OsEBF2were successfully constructed. Co-transform pGADT7-T/pGBKT7-53,pGADT7-OsEIL1/pGBKT7, pGADT7-OsEIL1/pGBKT7-OsEBF1,pGADT7-OsEIL1/pGBKT7-OsEBF2to the Ah109, using the nutrition defects medium SD/-Trp-Leu-His-Ade and the X-a-gal color experiment, it prove that the pGADT7-OsEILl/pGBKT7-OsEBFland pGADT7-OsEIL1/pGBKT7-OsEBF2bacterial coloneise grow appear blue the same as the positive control, which indicate that the OsEBF1and OsEBF2both interact with OsEIL1in yeast two-hybrid system.(2) For further illustrate the interactions between OsEIL1and OsEBF1,OsEIL1and OsEBf2completely, the sudy intent to use GST pull-down determine interactions. Another three GST pull-down recombinant vector, pGEX-6p-1-OsEIL1,pET-30a-OsEBF1,pET-30a-OsEBF2, were successfully constructed. These tasks lay a solid foundation for further research on the interaction between OsEILl and OsEBF1, OsEBF2in rice by GST pull-down.