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7d可这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.
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