摘要
用生物技术手段分离鹧鸪菜(Caloglossa leprieurii)原生质体,筛选出成功的分离条件。原生质体的活性用伊文斯蓝确定,原生质体及其去壁情况用荧光增白剂、低渗、自融和电镜4种力法观察、确定。原生质体成活率主要受种藻健康状况、酶解时间和培养液组成影响。细胞分裂频率与激素相关。在合适培养条件下获得较高的再生频率,并培养成正常植株,并且产生了无性生殖器——四分孢子囊。
Protoplast, as an important medium in genetic engineering, have been isolated from some genera of seaweeds in the near decade. Using the combinative enzyme of 3% Sea snail enzyme, 3% Cellulase Onozuka R-10, 1% Macerozyme and 0.5% Pectinase, viable protoplasts were isolated from Calogtossa leprieurii (Mont.) J. Ag. (Ceramiales, Rhodophyta). The protoplast viability was confirmed by exclusion of Evans Blue stain. The protoplasts and its absence of cell well were observed and confirmed by Calofluor White, reduced osmolarity, electron microscopic examination and self-fusion. About half number of protoplasts survived and divided with high ratio when cultivated in thin liquid layer of modified medium No.5 (MES as basic medium, supplemented with MS medium vitamins and 1ppm IAA, 1.5ppm KT). Regenerated plantlet appeared after 45 days. Tetrasporangia appeared in the blade of some regenerated plants after 4 months.
Being big, polynueleal and regenerative, protoplasts of C. leprieurii show prospect in the field of cell fusion and genetic operation of seaweeds.
出处
《生物工程学报》
CAS
CSCD
北大核心
1989年第4期297-302,共6页
Chinese Journal of Biotechnology
关键词
鹧鸪菜
原生质体
培养
Caloglossa
enzyme
protoplast culture
plant regeneration
