摘要
以‘富士’苹果(Malus×domestica Borkh.‘Fuji’)成熟花粉为试材,采用"萌发—酶解二步法",初步探讨了影响原生质体分离的关键因子,获得了花粉原生质体分离的最佳条件:用萌发后45 min的花粉,转入含1%纤维素酶和1%离析酶的混合酶液中,以18%甘露醇调节渗透压,静置酶解6 h,花粉原生质体分离效率可达6.83%,用0.1%荧光增白剂检测表明脱壁完全,用0.01%FDA检测表明其具有生活力,可以作为后续细胞融合等的材料。
Little information is available about the research on plant pollen protoplasts,especially the isolation of apple pollen protoplasts. In this study,the protoplasts from mature pollen of Malus × domestica Borkh.‘Fuji'by the method of‘germination-enzymatic treatment'were isolated. To screen the most effective condition of isolation,some key factors were analyzed. The results indicated that:The isolation percentage was up to 6.83% when the pollens germinating for 45 min and then treated with mixed enzyme solution including 1% Cellulase Onozuka R-10,1% Macerozyme R-10 and 18% Mannitol for 6 h. The cell walls of isolated protoplasts were degraded entirely according to the examination of 0.1% calcoflower white staining. 0.01% fluorescein diacetate(FDA)staining indicated the protoplasts were viable. Therefore,the proroplasts can be used for further research.
出处
《园艺学报》
CAS
CSCD
北大核心
2015年第6期1167-1174,共8页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(31171941
31372035)
关键词
苹果
花粉
原生质体
分离
apple
pollen
protoplast
isolation
