RNA Silencing Comparative Study Mediated By Tomato Yellow Leaf Curl Virus Six Functional Genes

Tomato yellow leaf curl virus (TYLCV) is a kind of geminivirus, spreading and diffusing depend on bemisia tabaci. In recent years, this kind of virus broke out in parts of tomato grown in China, serious prejudice and difficult to cure was the characteristics of this kind virus, has caused great harm to our country tomato production areas. With the development of RNA (RNA silencing) technology, It provide a new strategy for this disease prevention and control.This paper take TYLCCNV as research objects, design specific primers, AC1、 AC2、AC3、AC4、AV1、AV2six functional genes of TYLCCNV cloned by PCR. To copy these functional genes fragment by pEASY-T vector; Take PFGC-5941plasimd as objective vector, Linked functional genes fragment and PFGC-5941plasimd through four restriction enzymes named SmaI、XbaI、NcoI、AscI, builded the eukaryotic expression vector of inverted repeat sequence named AXF-PFGC-5941-AXR, test by PCR、digestion and sequencing, showed these functional genes fragment has been connected to PFGC-5941plasmid. Take the recombinant expression vector into agrobacterium LBA4404, Later take the recombinant expression vector into Nicotiana benthamiana through Agrobacturium-mediated leaf disk transformation method, Through kanamycin (Kan) and PCR checking, get a group of transgenic tobacco plants with resistance.Mediated by Agrobacterium tumefaciens inoculation method, proceed test to transgenic plant by toxicity attack, the experiment result indicated:these transgenic tobacco that contain AC1、AC3、AV1genes fragment respectively resistance ratio of83.3%、81.5%、89.6%, The three kinds of transgenic tobacco plants have high disease resistance.