In Vitro Expression Of Southern Rice Black-streaked Dwarf Virus P6 And Its Interaction With Antiviral Compounds

Southern rice black-streaked dwarf virus(SRBSDV)is belong to a reovirus family,Fijivirus.The SRBSDV is globular and contains a total of 10 RNA genes,of which the S6-encoded non-structural protein P6 is predicted to be a silencing suppressor.The silencing suppressor protein of the virus plays an important role in the viral infection cycle,and the viral silencing suppressor protein is widely used as a bait for screening host interaction factors.In the field,dufulin is a kind of drug with good control effect on southern rice black-streaked dwarf disease and to some certain extent,it controls a large-scale outbreak of southern rice black-streaked dwarf virus disease.In this study,the conditions of expression and purification of P6 protein in vitro were studied by molecular cloning and protein purification.The binding ability of antiviral agent duflin to SRBSDV P6 protein was studied by MST and ITC.In vitro,the effect of duflin on the expression of SRBSDV P6 protein in Nicotiana benthamiana was studied by Agrobacterium-mediated method.The effect of duflin on the expression of SRBSDV P6 protein in rice was studied in vivo.The results showed that the SRBSDV P6 protein with high purity was successfully purified by in vitro expression of SRBSDV P6 protein.The strong binding ability of duflin to P6 was studied by MST and ITC technology.The concentration of SRBSDV P6 protein decreased gradually,which revealed that the mechanism of action of duflin was to control the virulence of southern rice black-streaked dwarf virus by inhibiting the expression of P6 protein.