Remediation Of Soil Contaminated By Tobacco Major Viruses And Application Of Disinfectants

Tobacco virus disease is an important disease that harms the tobacco industry.Tobacco mosaic virus(TMV),Cucumber mosaic virus(CMV)and Potato virus Y(PVY)have brought serious economic losses to the tobacco industry.Virus infection at seeding period and soil poisoning in tobacco fields are the main causes of tobacco virus disease.Therefore,screening effective and safe disinfectants for major viruses and developing green remediation measures for toxic soils in tobacco fields are of great significance for the prevention and control of tobacco virus diseases and reduction of economic losses.In this study,eight kinds of chemical disinfectants were selected to disinfect the devices with TMV and CMV in the seed nursing period,and the best disinfectants for plant viruses were screened.At the same time,the use of solar energy disinfection,biological agents,and Chinese herbal medicine-tobacco intercropping were used to repair the virus-contaminated soil,and their inhibitory effects on the virus in the soil were analyzed.The main results are as follows:1.Using ELISA technology,8 kinds of chemical disinfectants were treated for disinfection of TMV and CMV.The results showed that the treatment with 2% of complex phenol for 30 minutes and 500 mg/L of sodium chlorite for 30 minutes had strong antigenic damage to both TMV and CMV.Yongjiekang disinfectant II(100 mg/L),povidone iodine solution(400 mg/L),2% glutaraldehyde disinfectant and sodium trichloroisocyanurate powder(800 mg/L)only for CMV The antigenic destructive effect reached 100%.2.Quantitative PCR was used to detect the range of gene of CP and Rep in TMV and CMV after treatment with composite sodium chlorite powder(500 mg/L).The results showed that the accumulation of four genes was reduced,and there was a significant difference with the positive control(P<0.01).3.Using large-step walking RT-PCR to detect the damage of the respective genomes after treatment of TMV and CMV with composite sodium chlorite powder(500 mg/L).The results show that the composite sodium chlorite first destroys the 183 kDa-2,183 kDa-4,and CP and MP regions of TMV and then disrupts the 183 kDa-1,183 kDa-3,and 54 kDa regions.First destroy the 1a-3 and 2b regions of CMV,then destroy the 1a-1,2a-1,and CP regions.Based on the results of ELISA,it can be determined that the composite sodium chlorite powder first destroys the coat protein of TMV and CMV and then destroys its internal genome.4.Observe the virus particles after treated with composite sodium chlorite powder(500mg/L)by electron microscopy.The observation results show that the rod-like granular structure of TMV is destroyed into pieces;the spherical structure of CMV has blurred edges.Viral bodies accumulate in clusters.5.Use of solar energy disinfection,biological agent spraying and medicinal plants-tobacco intercropping 3 ways to deal with virus soil.Quantitative PCR was used to detect the accumulation of gene of TMV CP in the leaves of cultivated tobacco after treatment.The results show that the solar disinfection treatment has a significant inhibitory effect on the virus in the soil.Spraying gallic acid,chitosan oligosaccharides and composite sodium chlorite separately in the poisoned soil can significantly inhibit the accumulation of gene of TMV CP in tobacco leaves.The accumulation of TMV in Radix Isatidis,Lithospermum erythrorhizon,Radix Glycyrrhizae and Sophora flavescens leaves and tobacco intercropping were significantly different from those of positive controls(P<0.05): they were reduced by 27.5,159.10,374.04,and 1161.14 folds,respectively.Among them,the anti-virus effect of Sophora flavescens is the most significant.