金花茶松散型愈伤组织诱导与悬浮细胞系建立

采用组织培养及显微观察技术,以金花茶植株茎段、叶片、花药及离体培养的金花茶体细胞胚为试验材料,对松散型愈伤组织的获得及悬浮细胞系的建立进行了研究。结果表明:在黑暗条件下,以金花茶体细胞胚切块为外植体,在MS+KT 0.5 mg/L+NAA 8.0 mg/L+硝酸银5mg/L+蔗糖30 g/L培养基上诱导出愈伤组织后,将其转接到该诱导培养基上连续培养3个月,最后从培养物中挑选状态良好的松散型愈伤组织在分别含有肌醇与硝酸银的2种培养基上交替培养,可以获得金花茶松散型愈伤组织;方差分析表明:蔗糖添加量、肌醇添加量及硝酸银添加量均对金花茶悬浮培养液细胞密度有显著影响且后两者分别与光照条件存在相互作用;将松散型愈伤组织在分别添加100 mg/L肌醇与2.5 mg/L硝酸银的液体增殖培养基上交替培养,可以建立并保持金花茶悬浮细胞系。该研究结果可为金花茶大规模细胞培养提供科学依据。

Induction of Friable Callus and the Establishment of Suspension Cell Lines ofCamellia nitidissima Chi

he stem, leaf, anther and somatic embryo of Camellia nitidissima Chi were used as the materials in this study. With the methods of tissue culture and microscopic observation, the induction of friable callus and establishment of suspension cell lines were conducted. The study showed the optimal process to obtain friable callus from C. nitidissima chi. According to this work, the slices of somatic embryo were cultured on the MS medium supplemented with 0.5 mg/L KT, 8.0 mg/L NAA, 5 mg/L AgNO3 and 30 g/L sucrose under dark condition to induce callus. The newly induced calli were separated from somatic embryo and were cultured on the callus inducing medium for three months. Finally, the friable calli were subcultured on the media containing inositol or AgNO3 alternatively; According to analysis of variance, the content of sucrose, inositol and AgNO3 significantly affected the cell density in suspension cultured liquid medium of C. nitidissima Chi and the lighting condition had significant interaction with the last two; The optimal process to establish suspension cell lines of C. nitidissima was also reported in this paper. In this process the friable callus were cultured alternatively in liquid proliferation medium containing 100 mg/L inositol or 2.5 mg/L AgNO3 under dark condition. This work can be basis for large-scale cell culture of C. nitidissima Chi.