Abstract: In order to improve the quality of fresh grape seedlings and the breeding speed of fresh varieties， the fresh grape variety Jinghongbao was used as the experimental material to explore the effects of factors such as the type and concentration of natural nutrient additives， disinfection time of explants， photoperiod， micro-grafting methods and binding materials on the survival rate of grape micro-grafted seedlings. The best experimental conditions for grape micro-grafted seedlings were determined by response surface methodology， and the grape micro-grafting tissue culture rapid propagation system was optimized. The results showed that the addition of banana mud in MS+0.2 mg/L IBA+1.0 mg/L 6-BA+0.2 mg/L NAA+30.0 g/L sucrose+7.0 g/L agar medium（pH 5.8～6.0） could significantly decrease the browning rate and pollution rate of micro-grafted seedlings and promote the elongation of adventitious roots. The response surface analysis further confirmed that the optimal experimental conditions for grape micro-grafting tissue culture were： HgCl2 disinfection time 7.1 min， light time 17 h， banana mud concentration 700 mg/L. On this basis， the survival rate of grape micro-grafted seedlings using the splicing method and tin foil as the binding material was significantly higher than other treatments.

Key words: grape, tissue culture, micrografting, response surface analysis