Detection And Molecular Identification Of Three Virus And Tomato Yellow Leaf Curl Virus Associated Satellite Infecting Solanaceae Vegetables

Plant virus is one of the most important negative factors in vegetables planting industry, plant viruses nowadays become the main restraining factor to sustainable development of vegetable industry.Tomato yellow leaf curl virus, TYLCV is a DNA virus from the genus "Begomovirus" and the family Geminiviridae,TYLCV can lead to devastating injury of vegetable industry by cause destructive Tomato yellow leaf curl disease through insect vector. Broad bean wilt virus, BBWV is a kind of pandemic virus in worldwide, it has wide host range and the infections and damages caused by this virus can be found in many vegetable planting areas. Tomato chlorosis virus, ToCV is a kind of newly found virus in China which spread and transmitted by whiteflies, it can infect a variety of plants.The main object of this research is to do investigations in Shandong, Henan, Hebei and Liaoning planting areas based on PCR technology about three viruses (BBWV, Tocv, and TYLCY), find out the infection areas and host range of BBWV, ToCV and TYLCY associate satellites.The research results showed that when TYLCV positive samples from Shandong, Henan, Hebei, Liaoning test with universal primers betaO1/betaO2 of betasatellite of geminivirus, approximately 1.3Kb specific bands of DNA?can be amplified in those susceptible samples from Shandong Changyi, Henan Jiaozuo. Through BLAST and the sequence comparison, the amplified sequence is 88% consistency with Vietnamese yellow leaf curl virus satellite DNA? (TYLCVNB). This result confirmed that TYLCV is associated betasatellite in tomato planting area in Shandong and Henan Province.This is also the first report of TYLCV viral DNA-A and DNA? (TYLCVNB) exist in complex forms.55 pepper samples were collected from Shandong province, using BBWVspecific primer pairs BBWV-R1F/R1R, R2F-1F/R2F-1R to do BBWV RT-PCR amplification, the results showed 570bp,1.3Kb,600bp,1.4Kb BBWV-RNA1 specific bands and two BBWV2-RNA2 specific bands contains SCP and LCP sequence were amplified, three positive samples screened out by the test, the results showed that the pepper samples collected from Shandong Changyi, Qingzhou was infected with BBWV. Wherein, BBWV2-RNA1 sequence and Korea isolate RP4 have a high sequence homology of 94.61%, consistency with the Korean isolate AF104335 and BBWV2-RNA2 is 96.03 percent.290 samples of tomato,48 weed samples were collected from 9 areas in HeBei province, and use three pairs of specific primers include ToCV coat protein and heat shock protein to test ToCV suspected samples, and amplified to about 1032bp,943bp and 911bp specific bands, found that 132 tomato samples were infected by ToCV, and none in the weeds samples. Then the ToCV positive samples were test with TYLCV specific primers, the amplified sequence is about 833bp, proved that is the exist of ToCV and TYLCV mixed infectionin fields. Comparison of Sequences obtained, ToCV sequence identity within isolates of Hebei is 99.7%, and is up 99.5% versus 99.7% with those isolates from Japan and Beijing.