葡萄愈伤组织制备原生质体的影响因素研究

【目的】为了获得高质量的葡萄原生质体,【方法】以"鄞红"葡萄茎段愈伤组织为材料,对葡萄原生质体制备过程中的若干影响因子,如酶解液成分、甘露醇浓度、酶解时间、离心条件等方面进行了研究。【结果】0.5 mol.L-1的甘露醇+1%纤维素酶+0.025%～0.05%果胶酶为最佳的酶解液组成;酶解14 h、120×g离心3 min条件下原生质体数量和质量最佳。【结论】研究获得的高质量葡萄原生质体,为下一步的植株再生及葡萄品种遗传改良提供了良好的材料。

Research on preparation method of protoplasts from callus of grape

【Objective】In order to obtain high quality protoplasts,【Method】the callus from stem of ‘Yinhong’ grape was used to study preparation method of protoplast.Some influence factors for isolation of protoplast,such as enzyme composition,concentration of mannitol in enzyme solution,duration of enzyme dissolve,and speed and minutes of centrifugation,and so on,were investigated and analyzed.【Result】The results showed that the optimal enzyme solution for protoplast isolation was 0.5 mol.L-1Mannitol + 1.0% Cellulase Onozuka R-10 + 0.025% to 0.05% Pectolyase Y-23;Highest yield and viability protoplasts were obtained when callus was incubated in enzyme dissolve for 14 h and centrifugated at 120×g for 3 min.【Conclusion】High quality protoplasts obtained in this study could be good materials for further research about plant regeneration and genetic improvement of grape.