The Interaction Between P6 Protein And Other Proteins Of Rice Black Streaked Dwarf Virus As Well As Proteins Of Oryza Sativa

Rice black streaked dwarf disease which is caused by Rice black streaked dwarf virus (RBSDV) is one of the most important viruses of rice and maize. Rice black streaked dwarf disease is widely spread in China. P6 encoded by RBSDV S6 were reported to be potentially as silencing suppressor.Fujian and Jiangsu isolates were amplified using RT-PCR, whose products were cloned into clone vector,which were transformed into Escherichia coli DH5αand were sent for sequencing. The sequencing results were analyzed and showed that the pathogen of Jiangsu isolates were RBSDV and Fujian isolates were mix infected by RBSDV and South rice black streaked dwarf virus (RBSDV). Yeast two-hybrid system were adopted on the study of P6. pGADT7-S5A,pGADT7-S8,pGADT7-S9-1,pGADT7-S9-2 and pGBKT7-S6 were constructed, by which open reading frames were cloned into yeast expression vectors pGADT7 and pGBKT7 respectively. It turned out that no interaction was detected between pGBKT7-S6 and pGADT7-S5A,pGADT7-S8,pGADT7-S9-1,pGADT7-S9-2, which meant that S6 and S5A,S8,S9-1,S9-2 might have no interaction.pGBKT7-S6 which was chosen as bait in yeast two-hybrid system was in order to screen rice seedling cDNA library. Four positive clones were screened named as SR-1,SR-2,SR-3 and SR-4, whose sequence were alignment with homologous genes in GenBank library or TIGR Rice Genome Annotation library. With the annotation of the libraries and studies of other researchers, the functions of SR-1,SR-2,SR-3 and SR-4 were identified as unknown,thylakoid-bound APX (tAPX),immutans protein and aldose 1-epimerase (A1EP) respectively. The four positive clones were preliminarily presumed to be enzymes in the synthesis processes of rice, which played an important role in the physiological processes of rice. Metabolism pathway of the hosts might be changed after the infection of RBSDV.