Construction Of Infectious Clone Of Tomato Yellow Leaf Curl Virus And Its Application In Tomato Breeding

In this study, molecular identification of TYLCV has been made with DNA of two disease samples from Handan. Then, we created infectious clone of the tomato yellow leaf curl virus. Using of infectious clones created, we explore a method of indoor artificial inoculation resistance identification. A pair of specific primers PA1-F/R has been designed in the conserved sequence, meanwhile PCR amplification has been made in DNA of two samples with diseases. Sequencing results of specific fragment 651bp is obtained from samples’amplification, based on its characteristics, a pair of forward primers RTF/R have been redesigned, and then from its amplification we get a full-length sequence of DNA-A of viral samples. Design specific primer PCR gene, gene cloning build, get to the purpose of the sequence after sequencing. Virus infection of cloning is by building a carrier, then transform the fruit of infectious clone to agrobacterium tumefaciens and make infectious experiment by the method of artificial inoculation. Identified by building and finally be required after infection sex cloning tomato disease research,The best time for vaccination is on the stage of seeding-two-leaf age; Secondly, the best vaccination method is microinjection; Moreover, the finest liquid spawn is sputum induced by AS; Finally, OD value control is around 1.5(with wavelength of 600mm). With this method of vaccination, plants will show disease symptom after the latency period of about 20d at the temperature of 25-28℃. Therefore,greater contributions can be expected on both TYLCV virus resistance breeding and TYLCV viral research in future.