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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

刘培, 卫志明, 许智宏, 褚晓明, 曹日强

中科院上海植物生理研究所植物分子遗传国家重点实验室.上海 200032；南京大学生物系.江苏,南京 210039

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44.06mg g~(-1)FW,而且其生产量在所测的80d内无明显下降。

关键词： 硬紫草; 原生质体; 克隆变异

Callus formation from cultured protoplasts of lithospermum erythrorhizon and variation of the somaclones obtained

Liu Pei, Wei Zhiming, Xu Zhihong, Zhu Xiaoming, Cao Riqiang

National Laboratory of Plant Molecular Genetics,Shanghai Institute of Plant Physiology,The Chinese Academy of Sciences.Shanghai 200032；China

Abstract

Cell line AR126 of Lithospermum erythrorhizon has been subcultured for 6 years and selected repeatedly by "small cell aggregates method", but the content of shikonin derivatives decreased rapidly with no selection. Protoplasts were enzymatically isolated from AR126 with a high protoplast yield of about 10~(6) g~(-1) FW. Only when protoplasts were cultured by agarose-liquid double layers method with glucose as osmoticum, were visible somaclones obtained. Among the numerous somaclones obtained, only 34 clones were chosen for the examination of growth rate and content of shikonin derivatives by twostage culture method. From the frequency distribution of somaclones with respect to growth rate at both growth and production stage and content of shikonin derivatives at production stage, it can be seen that it is possible to select for clones with both high growth rate and high content of shikonin derivatives. The production of shikonin derivatives of the best cell clone was 44.06 mg/g inoculum in 40 days which was almost 2.5 times that of the original cell line and kept stable during the 80 days under monitoring.

Key words: Lithospermum erythrorhizon;Protoplasts;Somaclonal variation

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引用本文：

刘培, 卫志明, 许智宏, 褚晓明, 曹日强. 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[J]. 生理学报 1996; 48 (3): .

Liu Pei, Wei Zhiming, Xu Zhihong, Zhu Xiaoming, Cao Riqiang. Callus formation from cultured protoplasts of lithospermum erythrorhizon and variation of the somaclones obtained. Acta Physiol Sin 1996; 48 (3): (in Chinese with English abstract).