Study On Increasing Tomato Anti - TMV Infection Ability By Using RNA Interference Technology

Tobacco mosaic virus is a RNA-virus, that infects plants, especially tobacco and other solanaceous plants, which serious threat to crop quality and yield. Methods to control widely used is breeding disease-resistant varieties, but it has some limitations. RNA interference technology as a new molecular biology technical has opened up a new road to controlling virus. On the basis of the RNA interference-mechanism, the sequence of TMV replicase-gene as been as the target sequence of RNAi in this experiment. The inverted repeat structure of target sequence was inserted into RNAi vector which containing 35 S super promoter. Than the RNAi expression vector infected into Agrobacterium tumefaciens by freeze thawing method, which infected tomato Sterile seedlings. based tissue culture we obtained three RNAi transgenic tomato which containing target sequence for RNAi. After Artificial inoculation TMV, The results showed that: compared with the normal control of tomato,there are two transgenic tomato which resisting TMV-virus. The total RNA of all samples has been extracted used for fluorescence quantitative PCR reaction. result analysis showed that: transgenic tomato shows resistance ability to TMV, the copy number of the replicase-gene m RNA in the process of growth was significantly lower than the other control tomato. the m RNA of replicase-gene were almost reduced to zero in the transgenic tomato which showing high resistance. This confirmed that the disease resistance of transgenic tomato was from the effect of RNAi, than RNAi has the application value in the virus disease prevention and control.