摘要
为探究适宜豆腐柴(Premna microphylla Turcz)细胞悬浮培养条件,以豆腐柴叶片为材料,进行了愈伤组织的诱导、继代培养、愈伤组织分散、悬浮培养的研究,建立了摇瓶悬浮培养体系。结果表明,豆腐柴叶片诱导得到的愈伤组织形态各异,其中诱导最适合悬浮培养的松散型胚性愈伤组织的培养基为MS+0.4 mg/L 2,4-D+0.4 mg/L NAA+3%蔗糖+0.8%琼脂。通过纤维素酶和果胶酶分散愈伤组织块可以得到较好的分散细胞系,最适合悬浮细胞生长的培养基条件为MS+0.4 mg/L 2,4-D+0.8 mg/L KT+3%蔗糖,其悬浮培养细胞鲜重增加量平均每7 d可达6.46倍,暗培养相对更适合细胞的增殖。经最佳培养条件下培养21 d的豆腐柴悬浮细胞中果胶和蛋白质平均含量达6.57%和0.12%,远低于豆腐柴天然叶片中的含量。
In order to find the suspension culture conditions of Premna microphylla Turca,the leaf blades of the experimented Premna microphylla Turca,induced embryogenic callus and subculture,which established a good dispersion and a high dynamic suspension cell line. The results showed that the callus induction from leaves were different. Which induces the most suitable medium for suspension culture of loose embryogenic callus contained MS＋0.4 mg/L 2,4-D＋0.4 mg/L NAA＋3% sucrose＋0.8%agar. Cell lines can be dispersed better by cellulase and pectinase dispersed callus block. The most suitable medium for suspension cell growth contains MS＋0.4 mg/L 2,4-D＋0.8 mg/L KT＋3% sucrose. And in this medium,the fresh weight of the suspension cultured cells could be increased by 6.46 times per 7 days. Dark culture was more suitable for cell proliferation. The average contents of pectin and protein were 6.57% and 0.12% in the suspended cells of Premna microphylla Turca,which had been cultured for 3 weeks under the optimum culture conditions.
出处
《湖北农业科学》
2017年第1期171-174,177,共5页
Hubei Agricultural Sciences
基金
湖北省自然科学基金计划项目(2008CDZ085)
