硬紫草悬浮细胞原生质体培养形成愈伤组织及其体细胞克隆变异  

Callus Formation from Cultured Protoplasts of Lithospermum erythrorhizon and Variation of the Somaclones Obtained

作　　者：刘培[1] 卫志明[1] 许智宏[1] 褚晓明[1] 曹日强[1] 

机构地区：[1]中国科学院上海植物生理研究所植物分子遗传国家重点实验室,南京大学生物系

出　　处：《植物生理学报（0257-4829)》1996年第3期243-250,共8页Acta Phytophysiologica Sinica

摘　　要：从硬紫草悬浮细胞系ＡＲ１２６分离原生质体，只有用葡萄糖作渗透剂经琼脂糖－液体双层培养才能获得可见的原生质体克隆，从中选择３４个克隆，用两阶段法生产紫草素及其衍生物，测量了它们在两阶段的细胞生长量及生产阶段的色素含量，并比较了其分布规律，其中最好的原生质体克隆色素生产量是起始悬浮系的２．５４倍，经培养４０ｄ达４４．０６ｍｇｇ－１ＦＷ，而且其生产量在所测的８０ｄ内无明显下降。Cell line AR126 of Lithospermum has been subcultured for 6years and selected repeatedly by 'smallcell aggregates method',but the contentof shikonin derivatives decreased rapidly with no selection. Protoplasts wereenzymatically isolated from AR126with a high protoplast yield of about 106g-1 FW. Only when protoplasts werecultured by agarose-liquid double layersmethod with glucose as osmoticum,were visible somaclones obtained.Among the numerous somaclonesobtained, only 34 clones were chosenfor the examination of growth rate andcontent of shikonin derivatives by twostuge culture method. From the frequency distribution of somaclones withrespect to growth rate at both growthand production sarge and content ofshikonin derivatives at productionsarge,it can be seen that it is possible toselect for clones with both high growthrate and high content of shikoninderivatives. The production of shikoninderivatives of the best cell clone was44. 06 mg/g inoculum in 40 dayswhich was almost 2. 5 times that of theoriginal cell line and kept stable duringthe 80 days under monitoring.

关 键 词：硬紫草 原生质体培养 克隆变异 体细胞 

分 类 号：Q949.777.4[生物学—植物学]