Screening Of Rice Host Factors Interacting With The P6 Of Rice Black-streaked Dwarf Virus And White-backed Planthopper Vector Factors Interacting With The CP Of Southern Rice Black-streaked Dwarf Virus

Rice viruses, one of the most destructive pathogens in rice production, are prevalent in rice growing areas in China and cause great economic losses in rice production. To elucidate the mechanism of interaction between virus and host and vector, the silencing suppressor protein P6 of Rice black-streaked dwarf virus (RBSDV) and the coat protein (CP) of Southern rice black-streaked dwarf virus (SRBSDV) were respectively used as baits to screen the cDNA libraries of rice and white-backed planthopper, and some rice host and white-backed planthopper vector interaction factors were obtained in this study.1 Screening of rice host factors interacting with RBSDV P6The recombination plasmid pGBKT7-P6 (RBSDV) used as a bait for yeast two-hybride assay was constructed, and confirmed that RBSDV P6 had no toxicity and no self-activated activity in yeast. Then the plasmid pGBKT7-P6 and rice cDNA library plasmid were co-transformed into yeast strain AH109. After screened on the SD/-His/-Leu/-Trp and SD/-His/-Leu/-Trp/-Ade mediums, eight different rice protein segments interacting with the RBSDV P6 in yeast were obtained. These candidates were listed as a protein containing the PHD-type domain, aldehyde dehydrogenase, COP9 signalosome complex subunit 5b, an unknown function protein (named as P6-100), a protein containing the conserved SNase (Staphylococcal nuclease) domain, putative ATP synthase delta chain, Ubiquitin-conjugating enzyme E2 (catalytic) and UDP-glucuronic acid decarboxylase. The full-length gene of P6-100 from rice was cloned and the interaction between P6-100 protein and RBSDV P6 was further confirmed by yeast two-hybrid assay and bimolecular fluorescence complementation experiment. These preliminary works lay a foundation for further understanding the mechanism of interaction between RBSDV and rice host.2 Screening of vector factors interacting with SRBSDV CPThe recombination plasmid pGBKT7-CP (SRBSDV) used as a bait for yeast two-hybrid assay was constructed, and confirmed that SRBSDV CP had no toxicity and no self-activated activity in yeast. Then the plasmid pGBKT7-CP and white-backed planthopper cDNA library plasmid were co-transformed into yeast strain AH109. After screened on the SD/-His/-Leu/-Trp and SD/-His/-Leu/-Trp/-Ade mediums, six different protein segments interacting with the SRBSDV CP were obtained. These candidates were listed as actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH3), the coat protein 2 (CP2) of Himetobi P virus (HIPV), a mitochondrial protein, GMP synthase and TCP 1 subunit gamma. The above preliminary results will help us to further understand the mechanism of virus-vector interaction and control Southern rice black-streaked dwarf virus disease.