Brucea Javanica (L.) Merr Extract,Ailanthus Altissima Extract And Bruticine D To TMV,CMV-RNA Inhibit Action

Tobacco mosaic virus is distributed in the world. It can infect many kinds of plants of Cruciferae,Solanaceae,Compositae, is one of economical important plant viruses. Cucumber mosaic virus's host is very widespread, can infect 85 families 1000 kind plants. TMV and CMV can compound infection. They not only cause the plant quality poor but create the huge economic loss. TMV,CMV has many kinds host, the strong resistance and the great harm in the production.Brucea javanica (L.)Merr and Ailanthus altissima Swingle belongs simaroubaceae, is used in Chinese traditional medicine. In formerly research of our laboratory, we get to know, within 6-8 hours after inoculation, the inhibitory rate of Brucea javanica (L.)Merr extract and Ailanthus altissima Swingle extract (1mg/mL) was 90.9% and 89.4%. Antiviral activity of Bruticine D against TMV infection and replication was screened by means of local lesion and leaf discs assay, the results demonstrated that the inhibitory TMV infection rate of Y-D (0.025mg/mL) was above 75%, when tested at a concentration of 2×10^-3 mg/mL, the inhibitory TMV replication rate of Y-D was 60%.Bruticine D,Brucea javanica (L.)Merr extract and Ailanthus altissima Swingle extract can inhibit TMV infection. To research the effect of TMV RNA,CMV RNA replication in vivo, we using semi-quantitive RT-PCR, choose the suitable annealing temperature, Mg²⁺concentration, the number of cycles of TMV replicase gene,coating protein gene(CP),movement protein gene(MP),tabacco actin gene and CMV-RNA3 . Further research that the K₃₂₆ within 6h after inoculation TMV and Nicandra physalodes (L.) Gaertner inoculation within 12h after inoculation CMV, with Bruticine D,Brucea javanica (L.)Merr extract and Ailanthus altissima Swingle extract soaking the inoculated leaves for 48h, detect the three medicine inhibitory TMV-RNA,CMV-RNA replication. We also use quantitive real-time PCR to detect Y-D inhibitory TMV-RNA,CMV-RNA replication, the inhibitory rate to TMV replicase gene, TMV-CP,TMV-MP is 93.04%, 94.79% and 95.82%. The results of the two methods are the same, They can effectively inhibit TMV-RNA replication. We also get to know the result of semi-quantitative RT-PCR is correct, and confirm semi-quantitative RT-PCR Reliability. Because of the quantitive real-time PCR instrument is expensive, when the condition is limited. If operation appropriated, semi-quantitative RT-PCR can displace quantitive real-time PCR. From the experiment result, the medicine almost can not inhibit CMV-RNA replication. But we can not get the conclusion the medicine can not inhibit CMV-RNA replication. And also can not conclude the three medicine can't generally inhibit +ssRNA virus replication. We can futher research inhibit virus protein.