Study On The Interaction Between Rice Ethylene Receptors And OsTPR1 Protein

Plant TPR proteins (AtTPR1 and S1TPR1) can interact with the specific ethylene receptors. And this interaction plays a very important role in plant ethylene signaling and the growth and development of plants. OsTPR1 is a rice TPR protein, which also may interacts with the specific rice ethylene receptors. And this interaction also may play a role in rice ethylene signaling and influence rice growth and development.In this study, the interactions between rice ethylene receptors and OsTPR1 were examined via yeast two-hybrid system and bimolecular fluorescence complementation (BiFC), the specific domains of their interactions were further identified. The main research results are as follows:(1) One bait plasmid pGBKT7-OsTPR1 and three yeast expression vetors of rice ethylene receptors including pGADT7-OsERS1,pGADT7-OsERS2,pGADT7-OsETR2 were successfully constructed. Only OsERS1 was found to interact with OsTPR1 in yeast two-hybrid system.(2) To future identify the specific domain of OsERS1 that interacts with OsTPRl, we used bioinformatics tools to analyze the structure domain information about OsERS1. Based on this information, four yeast expression vectors containing different domains of OsERS1 were constructed:pGADT7-OsERS1-1,pGADT7-OsERS1-2,pGADT7-OsERS1-3 and pGADT7-OsERS1-4. Yeast two-hybrid assay shows the specific interaction domains of OsERS1 were histidine kinase domain(OsERS1-3) and histidine kinase-like ATPases_c domain(OsERSl-4). These results provide a forceful support for further research on the influence of OsTPR1 on OsERS1.(3)Another two BiFC recombinant vector, pUC-SPYNE-OsERS1 and pUC-SPYCE-OsTPR1, were successfully constructed. These tasks lay a solid foundation for further research on the interaction between OsERS1 and OsTPR1 in rice protoplasts by BiFC.