欧白英悬浮细胞原生质体培养和植株再生  

Culture and Regeneration of Solanum dulcamara L.Protoplasts Derived from Cell Suspension

作　　者：王国英[1] 

机构地区：[1]北京农业大学生物学院

出　　处：《农业生物技术学报》1994年第1期64-69,共6页Journal of Agricultural Biotechnology

摘　　要：本文报道了欧白英细胞悬浮系的建立和原生质体培养技术。从欧白英悬浮细胞分离原生质体，以含２％ＲｈｏｚｙｍｅＨＰ１５０，２％ＭｅｉｃｅｌａｓｅＣＥＳＢ和０．０３％ＭａｃｅｒｏｚｙｍｅＲ－１０的混合酶液效果最好，平均每克细胞团可得到２．０５×１０６个原生质体。悬浮系在继代培养第４天分离的原生质体植板率最高，在液体培养基中原生质体接种密度以１．５×１０５个·ｍＬ－１最佳，植扳率可达到２４．４３％。培养５０天后，约有０．２１％的原生质体形成１～２ｍｍ大的细胞克隆。将它们转到Ｋ８培养基上，形成愈伤组织块。再转入ＭＳＺ培养基，２～４周即有３０％～４０％分化出不定芽，长出枝条。再将幼枝转入不含激素的ＭＳ培养基，２周后大部分枝条生根，长成植株。This paper reported the establishment of S.dulcamara L. cell suspension and the culture ofsuspention-derived protoplasts. The results showed that young leaves could produce more T and TG types ofcalli,which grew rapidly during subculture and later were apt to form lcose yellow calli and suitable for estab-lishment of cell suspension. The maximum amount of protoplasts2.05 × 106 protoplasts/gf.wt,was ob-tained by using the enzyme mixture consisting of 2.0%Rhozyme HP 150,2.0%Meicelase CESB and 0.03%Macerozyme R-10 in CPW 13M at pH 5.8. The experimental results also showed that the highest plating efficiency could be obtained from protoplas-ts isolated at the logarithmic growth phase(4 days after subculture).The optimum plating density of proto-plasts was 1.5 × 105/mL with a plating efficiency of 24.43%in liquid culture. Under the optimum cultureconditions,about 0.21%of protoplasts formed 1～2 mm microcolonies after 50 days of culture,which grewrapidly after being transferred to K8 medium with 0.6%of agar. Adventitious shcots were regenerated fromabout 30%～40%of the calli on MSZ medium after 2～4weeks of culture. Most of the shcots rooted on MSmedlum without the suppliment of plant growth regulators after 2 weeks of culture,

关 键 词：药用植物 欧白英 原生质体 培养 植株再生 

分 类 号：S567.239[农业科学—中草药栽培]