| In this experiment,three new strains wine grape ’Beichun’ ’ Beichun 11’,’ Beichun 12’ and’Xinbeichun’ were used as experimental materials to explore the content of common viruses in different parts of the plant and establish virus-free and rapid propagation system for these three strains.The results showed that:1.RT-qPCR technique was used to detect different parts of three strains of plants selected for pathogen identification.The tested samples contained glrav-1,GLrav-2,GLrav-3,GLrav-4,GLrav-7,GVA,GVB,GVE,GFabV,GRSPaV,GPGV,GFkV and other 12 viruses.Different parts of the same strain have different virus species and content.There were significant differences in virus content in different parts of Beichun 11’.Among the 12 detected viruses,the whole plant was infected with different relative expression levels of viruses.Petiole had the most virus species which contains 10 kinds;New shoot contains 9 kinds of viruses,take the second place;The least species of viruses are leaves,which contain only two viruses,GRSPaV and GVE.Among the 12 detected viruses,’Xinbeichun’ has the most kinds of viruses in leaves,including 8 kinds.Followed by the root system,there are 6 kinds of viruses in the root system and 4 viruses in the branches.2.In the process of tissue culture of ’Beichun 11’,’Beichun 12’ and ’Xinbeichun’,different sampling time,disinfection methods and media with different hormone concentrations have different effects on the growth of explants.In the primary culture,the combination of alcohol disinfection for 1min and 15%hydrogen peroxide disinfection for 6min had the best disinfection effect.The best start-up medium for the three strains was MS medium+1.0 mg·L-1 6-BA+0.3 mg·L-1 NAA.The optimum medium for shoot tip culture is 1/2MS medium+2.0 mg·L-16-BA mg·L-1+0.1 mg·L-1NAA.The combination of NAA and IBA in rooting medium has a significant effect on rooting index,and the optimum hormone concentration is 1/2MS medium+0.2 mg·L-1IBA mg·L-1+0.4 mg·L-1NAA.3.In the process of detoxification,the combination of shoot tip culture and drug treatment was used to obtain better detoxification effect.After using shoot tip culture and 15mg/L ribavirin detoxification,the final detoxification rate of tested plants reached 100%,all viruses were eliminated.In the plants detected by shoot tip culture and 3g/L Radix Isatidis detoxification,the GVB virus in ’Beichun 11’ and GVE virus,GFkV virus in ’Beichun 12’ were not removed,all the viruses detected in ’Xinbeichun’ strain were removed. |
