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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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Citing This Article：

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).