| In this study,eight year old ’Beixi’,"BeiXiang’ and ’Beixin’ wine grape varieties in the eastern foot of Helan,Ningxia,were selected as the research objects."Beixi"," Beixiang " and "Beixin" are three new varieties of "beizihao" introduced in Ningxia in recent years.At present,relevant symptoms of grape leaf roll disease,grape spot disease and other viruses have appeared in the cultivation in Ningxia,such as yellow and red leaves,transparent spots near leaf veins and other symptoms.Therefore,in this study,fluorescence quantitative PCR was used to detect the virus infection in the six parts of the stem tip,tender leaf,old leaf,petiole,branch and root of the three varieties of ’Beixi’,’Beixiang’ and ’Beixin’,to explore the tissue culture rapid propagation lines suitable for the three varieties of Beixi’,’ Beixiang ’ and ’Beixin’,and to screen the appropriate virus-free treatment methods,The main results are as follows:1.The virus infection of 12 kinds of viruses in different parts of "Beixi","BEIXIANG" and "Beixin"was investigated.A total of 10 kinds of viruses were detected in the three varieties,of which " Beixiang "had the most types of viruses,with a total of 9 kinds,They are GLRaV-2,Glrav-3,Glrav-4,Glrav-7,GFkV,GVA,GVB,GVE and GFabv respectively.There are many kinds of viruses in leaves.Eight species were detected in ’Beixin’,which were grape leaf roll associated with type 3 virus,GVB,GVA,glrav-4,GFabv,GVE,GPGV and GFkV.There were many kinds of viruses in stem tip and root’ Four kinds of GVB,Glrav-7,Glrav-4 and GFkV were detected in Beixi ’,and the virus was mainly distributed in the stem tip.There were 4 species of rav-glv in the stem tip and GVB.2.Taking the stem segments of ’Beixi’,’BEIXIANG’ and ’Beixin’ taken in the field as the initial explants,the key steps such as disinfection method,initial medium,subculture medium and rooting medium of stem segments of different varieties were studied.The results showed that the most suitable disinfection method of Beixin’ and ’Beixi’ was 70%ethanol 50(s)+15%H2O2 for 6min,the best disinfection method of’Beixiang’ was 70%ethanol50(s)+15%H2O2 for 5min,the best initial medium of the three varieties was MS+1 mg/L 6-BA+0.3 mg/L NAA,and the best subculture medium of’Beixin’and ’Beixi’ was 1/2 MS+ 2 mg/L 6-BA+0.1 mg/L,1/2 MS+2 mg/L 6-BA+0.1 mg/L NAA,The subculture medium of ’Beixiang’ was 1/2 MS+2 mg/L6-BA+0.2 mg/L NAA.The rooting medium selected by the three varieties was 1/2 MS+0.4 mg/L IB A+0.1 mg/L NAA.3.Taking the aseptic tissue culture seedlings of ’Beixi’,’Beixin’ and ’Beixiang’ as test materials,the aseptic tissue culture seedlings were treated with two detoxification methods:stem tip culture detoxification and stem tip combined with medicament culture detoxification.The research showed that the best detoxification method of’Beixin’ and ’Beixiang’ was 0.8 mm,the stem tip combined with 20 mg/L Virazole solution,the regeneration rate was 43.3%and 57%respectively,all viruses detected by ’Beixin’were removed,and glrav-7 and GFkV in BEIXIANG’ were not removed.and the regeneration rate of’Beixi’ was 0.8 mm,and the stem tip combined with 10 mg/L Virazole solution,the regeneration rate was 66.66%,Virus removal rate of all viruses detected by "Beixin". |
