摘要
以‘过山香’香蕉的胚性悬浮细胞(Embryogenic cell suspensions,ECS)为起始材料分离原生质体,酶解液的组成为:3.5%纤维素酶R-10、1%离析酶R-10、0.15%果胶酶Y-23、204mmol·L-1KCl、67mmol·L-1CaCl2和0.41mol·L-1甘露醇,原生质体产量为3.1×107.mL-1PCVECS(PCV:packed cell volume,细胞密实体积)。分别以培养基‘A’和‘B’为培养成分在液体浅层培养和看护培养两种培养系统中进行原生质体培养。结果表明:在液体浅层培养系统中,采用培养基‘B’比培养基‘A’效果好,原生质体的细胞分裂频率和细胞团形成频率分别约是采用培养基‘A’的3倍和10倍;所获得的培养物为只能增殖而不能进一步分化的非胚性细胞团。在看护培养系统中,采用培养基‘A’与培养基‘B’时,细胞分裂频率和细胞团形成频率没有显著差异;所获得的细胞团具有典型的胚性细胞特征。将从看护培养中获得的细胞团转移到体胚诱导培养基上,培养45d后,从105个原生质体获得1550个体胚。继续在体胚诱导培养基上培养30d,7.8%的体胚能萌发。萌发的体胚在MS+0.1%活性炭的培养基中发育成健壮植株,移栽后成活良好。
A protocol for plant regeneration from protoplast of Musa AAB Silk ＇Guoshanxiang＇ via somatic embryogenesis was developed.Viable protoplasts were isolated from embryogenic cell suspension（ECS）in a enzyme mixture of 3.5% cellulose R-10,1% macerozyme R-10,0.15% pectinase Y-23 and 0.41 mol·L^-1 manntitol,the yield was 3.1×10^7 protoplasts per mL packed cell volume（PCV）ECS.Liquid and feeder layer culture systems with medium ＇A＇ and medium ＇B＇ were used respectively for protoplast culture.In liquid culture system,medium ＇B＇ was more efficient for inducing cell division and colony formation which was about 3-fold and 10-fold respectively,compared to that with medium ＇A＇.However,all protoplast-derived cell colonies obtained from liquid culture system could not develop further.In feeder layer culture system,there was no significant difference between medium ＇A＇ and medium ＇B＇ on cell division and colony formation of the cultured protoplasts.Protoplast-derived cell colonies from feeder layer culture system were then transferred onto embryo induction medium for somatic embryogenesis.After forty-five days the cell colonies were transferred on embryo induction medium,1 550 mature embryos were obtained from 105 protoplasts.After another 30 d of culture,7.8% of mature embryos germinated.Normal plantlets were obtained from MS basal medium supplemented with 0.1% activated charcoal and the plantlets were transferred into pots and grew well.
出处
《园艺学报》
CAS
CSCD
北大核心
2008年第6期873-878,共6页
Acta Horticulturae Sinica
基金
国家青年自然科学基金项目(30400287)
广东省自然科学基金博士启动项目(04300538)
广东省自然科学基金项目(06023159)
广东省科技攻关项目(2006B20101014)
广州市科技计划项目(2006Z3-E0281)
关键词
香蕉
胚性悬浮细胞
原生质体
植株再生
banana
protoplast
somatic embryogenesis
plant regeneration
