Analyzing The Interaction Between Tomato Mosaic Virus Coat Protein And Tobacco IP-L

Tomato mosaic virus (ToMV), a member of the Tobamovirus genus, has a positive-sense single-stranded RNA genome of about6400nucleotides in length, which encodes three non-structural proteins and a17.6-kDa coat protein (CP). ToMV can infect tomato via juice and seed dissemination, causing severe influence on the quality and yield of tomato. Over the past decade, researchers conducted a variety of studies to investigate the biologic and molecular biologic characteristics of ToMV and have accumulated abundant information for the prevention and control of this virus. However, the underlying interaction mechanism between ToMV and host plants was not completely understood. Previous work showed that CP-interacting protein-L (IP-L) can interact in vitro with ToMV CP. In view of this point, the interactions of ToMV CP with Tobacco IP-L were investigated in this study to gain better understanding on the molecular pathogenic mechanism of ToMV.However, the underlying interaction mechanism between ToMV and host plants was not completely understood. Previous work showed that CP-interacting protein-L (IP-L) can interact in vitro with ToMV CP. What’s the relation between this interaction and yellowing. In view of this point, the mechanism of the interaction of ToMV CP with Tobacco IP-L has become a primary problemt. To adically explorer the significance of this interaction, the point is to study the interaction of two proteins. In this study we give the preliminary analysis for the interaction to gain better understanding on the molecular pathogenic mechanism of ToMV.Since continuously,the study for IP-L is only in the leaf of tobacco,but never in other components. Besides that, whether the IP-L exists in other Solanaceae plants.To conferm whether IP-L is only exsit in the leaf of tobacco and whether it exsit in other Solanaceae plants,we identify IP-L through RT-PCR.The rusuts shows that IP-L exsit in root, stem and leaf of tobacco, not only in the leaf,and it exsits in eggplants, peppers, potatos and tomatos,not only in tobaccos. The amino acid sequence comparison of IP-L in peppers, tomatos, eggplants and potatos with tobaccos are94.2%、94.7%、 95.3%、94.9%。Besides key area of ineraction, helix a,is identical with the amino acid sequence of IP-L in tobaccos.To further examine the interaction between ToMV CP and IP-L in tobacco body, three plant expression plasmids, i.e., pUC-SPYCE-35S-△IP-L、pSPYNE-35S-△CP106and pSPYNE-35S-△CP105were constructed. Bimolecular fluorescence complementation (BiFC) technique was used to detect the interaction of these plasmids with ToMV CP. The results showed that AIP-L can interact with ToMV CP in the living cells of tobacco leaves dispite the fluorescence is weaker than the fluorescence of IP-L and CP.△CP106and△CP105can interact with IP-L in the living cells of tobacco leaves. Thus△IP-L (L9A, H2G) is not the necessary part of interaction.△CP106and△CP105are necessary part of the interaction.