贡蕉胚性悬浮细胞原生质体分离的研究

Study on Protoplast Isolation of Embryogenic Cell Suspensions of Musa acuminata cv.Mas(AA)

目的:研究不同方法对贡蕉胚性悬浮细胞原生质体分离的影响,筛选适合用于贡蕉胚性悬浮细胞原生质体分离的方案。方法:用不同的酶浓度、酶组合及不同的酶解时间对贡蕉胚性悬浮细胞进行原生质体分离,并对不同继代时间的胚性悬浮细胞的原生质体产量和活力进行研究。结果:贡蕉胚性悬浮细胞在酶组合为3.5%纤维素酶R-10、1%离析酶R-10和0.15%果胶酶Y-23的酶溶液中,酶解8h可获得高产量的原生质体,采用继代7d的贡蕉胚性悬浮细胞进行原生质体分离时获得的原生质体产量最高,达到1.2×107个/mL PCV ECS,原生质体活力达到85%以上。结论:合适的酶组合、酶浓度和酶解时间有利于贡蕉胚性悬浮细胞的原生质体分离,继代7d后的贡蕉胚性悬浮细胞最适合用于原生质体分离。

Objective： To study the effects of differem methods on protoplast isolation of embryogenic cell suspensions of Musa acuminata cv. Mas （AA）. Method： Different concentrations and compositions of enzymes were used for protoplast isolation of embryogeneic cell suspensions （ECS） of Musa acuminata cv. Mas （AA）, and the yield of protoplasts isolated at various length of time after subculturing were studied. Result：The results indicated that enzyme mixture of 3.5% cellulose R- 10, 1% macerozyme R- 10 and 0.15% pectinase Y - 23 for 8 h were most efficient for protoplast isolation, highest yield of protoplast were obtained from 7 - day - old embryogenic cell suspensions （ECS） with a yield of 1.2 ×10^7 protoplasts/mL packed cell volume （PCV） ECS. Conclusion： Suitable concentration and composition of enzyme, exposure time were beneficial for protoplast isolation, embryogenic cell suspensions after 7 days of subculture were most efficient for protoplast isolation.