野生阿宽蕉胚性细胞悬浮系原生质体分离及植株再生

Protoplast Isolation from Somatic Embryogenic Cell Suspension Cultures of Musa itinerans Cheesman and Plant Regeneration

本研究以野生阿宽蕉(Musa itinerans Cheesman)未成熟种子诱导的胚性细胞悬浮系为材料,对其原生质体的分离和植株再生进行研究。结果表明:胚性悬浮细胞在酶组合:3.0%纤维素酶R-10、1.0%离析酶R-10、0.2%果胶酶Y-23、1.52%KCl、0.78%CaCl2、0.01%MES和11%甘露醇中,酶解8h,原生质体的产量和活力达到最大值,分别为19.46×106个/mL和92.17%。采用看护培养法对原生质体培养60d后,可获得大量增生的小细胞团,细胞团悬浮培养15d后转至体胚诱导培养基上培养约30d,其体胚萌发率为45.97%,然后转至生根培养基上培养35d,其再生植株率为54.76%。

In this research,we employed embryogenic cell suspension（ECS） induced from immature seeds of Musa itinerans Cheesman to be as materials for studying the protoplast isolation and plant regeneration.The results showed that enzyme concentrations of embryogenic suspension cell contained 3.0% cellulose Onozuka R-10,1.0% macerozyme R-10,0.2% pectinase Y-23,1.52% KCl,0.78% CaCl2,0.01% MES and 11% mannitol,and en zymes solution for 8 hours,then the yield and viability of protoplasts reached the highest,19.46×106 protoplasts/mL and 92.17%,respectively.Feeder layer culture systems were used for protoplast culture,and a great deal of cell aggregate clusters were obtained after 60 days culture,and then transferred onto embryo induction medium for somatic embryogenesis.30 days after cultured 15 days later,germination rate of somatic embryos was 45.97%,finally,the germinated somatic embryos were transferred to rooting medium for 35 days,the rate of plant regeneration reached 54.76%.