Function Of Pip2-1Gene Of Rice In The Process Of RSV Infection

Eukaryotic translation initiation factor is a kind of important proteins involved in proteintranslation initiation process. It plays key roles in the whole proteins translation process. Alarge number of studies have shown that many translation initiation factors are also involvedin the plant resistance pathway, besides protein translation initiation process.Eukaryotic translation initiation factor has a large protein family. The Pip2-1protein inthis study is one of the family members. In this work, we identified interaction betweenPip2-1protein and RSV P2protein through the Yeast two hybrid and Bimolecularfluorescence complementation experiments. Subcellular localization analysis showed that theprotein locates on the cell membrane. When we used virus induced gene silencing (VIGS)technique to induce Pip2-1gene silencing in plant.Plants showed dwarf, leaf malformation,blossom anomalies and low seed setting rate, which indicated that Pip2-1gene was essentialin the process of plant growth and development. The results of Real-time PCR showed thatthe Pip2-1gene expression level was down-regulated in RSV infected tobacco and rice plant,indicating that Pip2-1might have certain biological function in the process of RSV infection.These results provide clues for the RSV infection mechanism as well as the host resistancemechanism.In order to do further investigation of Pip2-1gene in the growth and developementprocess of Nicotiana benthamiana and rice as well as the relationship during virus infectioncycle, we cloned NbPip2-1and OsPip2-1gene sequences from Nicotiana benthamiana andrice, and constructed the over-expression vector and the inhibition expression vector ofNbPip2-1and OsPip2-1. By agrobacterium-mediated transformation, transgenic Nicotianabenthamiana and rice plants were obtained. Real-time PCR and Western blot showed that thePip2-1gene had been successfully integrated into the genome of transgenic plants as well asexpressed effectively. These results laid an important foundation for the later study, such asthe pHenotype analysis, function identification and resistance reseach of the transgenicplants.