Screening And Study Of Laodelphax Striatellus Proteins Interacting With Rice Stripe Tenuivirus Pc2

The pathogen of rice stripe disease is rice stripe tenuivirus(RSV).RSV is mainly transmitted by small brown planthopper(Laodelphax striatellus)in a persistent,propagative way.Once infected,the virus can be permanently carried and spread,which brings difficulties to viral prevention and control.Therefore,the research on RS V is conducive to the prevention and treatment of rice stripe disease.RSV genome can encode 7 proteins,among which there are few reports on Pc2 proteins.Pc2-N can be detected in both rice and insect infected by RSV,indicating that it may play a role in the process of virus transmission and inducion of symptom in host plants.In this paper,we studied the interaction of Pc2 with the protein in the small brown planthopper.In this paper,Laodelphax striatellus proteins interacted with RSV Pc2 was screened by yeast two-hybrid.In this section,the bait vector of the yeast two-hybrid nuclear system and the cDNA library of Laodelphax striatellus were respectively constructed and tested,indicating that they could be used for subsequent experiments.Five proteins that may interact with Pc2-N were screened,and one of them,Laodelphax striatella actin 1(LsATl)was selected for further study.And then the bait vector and the required cDNA library of Laodelphax striatellus were constructed respectively for the separation of ubiquitin-yeast two-hybrid system.After detection,three potential interacting proteins were obtained,and two potential interacting proteins LsSTl and LsHSC70 were selected for further study.The transmembrane domain and signal peptide analysis of the three possible interacting proteins showed that there are 12 transmembrane domains in the Laodelphax striatella sugar transporter 1(LsSTl),Laodelphax striatella actin 1 and Laodelphax striatella heat shock protein(LsHSC70)has no transmembrane region.After that,the interaction between Pc2-N and LsATl,LsST1 and LsHSC70 were further confirmed by the bimolecular fluorescent complimentaion and co-immunoprecipitation.The interaction between Pc2-N and LsAT1,Pc2-M2N and LsST1 was proved to be interaction with each other,and Pc2-M2N and LsHSC70 have a great possibility to be interaction with each other.Finally,we analyzed the distribution of the interacted proteins in the Sf9 cells by subcellular localization,and found that Pc2-N and LsAT1,Pc2-M2N and LsST1,Pc2-M2N and LsHSC70 had co-localization,and LsST1 changed the location of Pc2.Moreover,where there is little or no expression of LsST1,Pc2-M2N is overexpressed.These results showed that they might play a certain role in the mediator transmission of RSV,laying a foundation for further study of the mechanism of vector-mediated transmission.