The Research Of Replicase Gene Different Partial Sequences Mediates Resistance To Alfalfa Mosaic Virus

The full length of AlMV replicase gene cDNA sequence, as well as 168bp cDNA sequence at the 3'-end were respectively fused with the plant binary expression vector pROKII, correspondingly, the recombinated vectors pROKAMVF pR0KAMV3N were obtained, The half part of cDNA at 5'-end & 3'-end have been recombinated into binary vector pROK II during the former work of our lab, and recombinated vectors pR0KAMV5 pR0KAMV3 were obtained.According to this experiment, the four recombined expression vectors were mated into Agrobacterium tumefacient LBA4404 and then transformed into tobacco. PCR analysis was employed to detect the four ontogenious sequence of the gained transformants. The result show that 90 pROKAMV5 transgenic plants, 69 pROKAMV3 transgenic plants, 4 pROKAMVF transgenic plants 3 pROKAMV5 transgenic plants were obtained.Detected the transgenic plants by ELISA after inoculate AlMV 60 days, 10 thereinto 24 transgenic plants chains transformed with pROKAMV5 confer AlMV virus resistance, 12 thereinto 18 transgenic plants chains transformed with pROKAMV3 confer AlMV virus resistance.The research indicated that different sequence of AlMV replicase gene transformants perform resistance to virus infection in different level.