'过山香'香蕉胚性悬浮细胞原生质体分离的方法研究

Research on Protoplast Isolation of Embryogenic Cell Suspensions of Musa Silk cv. Guoshanxiang(AAB)

目的:研究不同的方法对'过山香'胚性悬浮细胞原生质体分离的影响,筛选最适合用于'过山香'香蕉胚性悬浮细胞原生质体分离的方案。方法:用不同浓度、不同组合的酶液对'过山香'原生质体进行分离,并对酶液的甘露醇含量、pH值进行调节。结果:3.0%纤维素酶R-10+0.2%果胶酶Y-23的是最佳酶组合;酶解8h、酶液中含0.41M甘露醇、酶液pH值为5.3时,获得原生质体产量最高。结论:合适的酶组合、酶解时间、酶液的渗透压和pH值对'过山香'香蕉胚性悬浮细胞原生质体的分离有明显的促进作用。

Objective： To study the effects of different methods on protoplast isolation of embryogenic cell suspensions of Musa Silk cv. Guoshanxiang （AAB）. Methods： Different enzyme concentrations, compositions, osmotic press and pH value of enzyme solution were used and adjusted for protoplast isolation of Musa Silk cv. Guoshanxiang （AAB）. Results： The results indicated that embryogenic cell suspensions were most efficient for protoplast isolation in the condition of enzyme mixture of 3.0 % cellulose R-10 and 0.20% pecti- nase Y-23 for 8 h. Enzyme solution with 0.41 mol/L manntitol and pH 5.3 resulted in the highest yield of 1.2 ×10^7 protoplasts/mL packed cell volume （PCV） ECS. Conclusions： Suitable concentration and composition of enzyme, ostimic press and pH value of enzyme solution were important for protoplast isolation of Musa Silk cv. Guoshanxiang （AAB）.