响应面法优化葡萄微嫁接组培快繁体系

Optimization of grape micro-grafting tissue culture and rapid propagation system by response surface methodology

为提高鲜食葡萄苗木品质和良种繁育速度,以鲜食葡萄品种晶红宝为试验材料,探究培养基天然营养添加物的种类及浓度、外植体消毒时间、光周期、微嫁接方法和绑缚材料等因素对葡萄微嫁接苗成活率的影响,结合响应面法确定葡萄微嫁接苗组培最佳试验条件,优化葡萄微嫁接组培快繁体系。结果表明,MS+0.2 mg/L IBA+1.0 mg/L 6-BA+0.2 mg/L NAA+30.0 g/L蔗糖+7.0 g/L琼脂培养基(pH 5.8～6.0)中添加香蕉泥可显著降低微嫁接苗的褐变率和污染率,促进不定根的伸长。响应面分析进一步确定了葡萄微嫁接组培的最佳试验条件为氯化汞消毒7.1 min,光照17 h,香蕉泥浓度700 mg/L。在此基础上采用劈接法、锡箔纸作绑缚材料的葡萄微嫁接苗成活率显著高于其他处理。

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.