Regulation of Vitamin E Production in the Revised Cell Line of Safflower

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A better cell line, Caf -B2KC strain, in terms of both the growth rate and amounts of tocopherols was obtained by selection using various growth regulators and medium additives. In addition, the tocopherol production was stimulated by administration of biosynthetic precursors, especially phytol, which is a precursor of the side chain in tocopherol. The total tocopherol content increased by some 5-fold. Moreover, a light green strain, named Ca- 2, which was isolated from the seedling under illumination, produced by 1.4-fold in the total tocopherol in comparison with that of Caf-B2KC. However, the Ca-2 strain never became further dark green by any treatment and the tocopherol content also did not increase.The main object in this research was the increase of tocopherol content in safflower, Carthamus tinctorius, cultured cells by greening by means of the following 3 methods. (1 )Electric cell fusion between Caf -B2KC callus and mesophyll cell of original plant via every protoplasts using … More Electric cell fusion apparatus. (2)Introduction of chloroplasts isolated from leaf mesophyll into Caf-B2KC callus by an electroporation method (3)Injection of chloroplasts to Caf-B2KC callus by a microinjection method. The protoplasts were formed in a high yield from both the cultured cells and leaf mesophyll cells, but they were very unstable for physical change, such as osmotic pressure and the other shocks. Therefore, the dark green callus containing many chloroplasts could not be formed by above three methods, even if the protoplasts were isolated from leaf mesophyll of original plant.A new green strain Ca-3, which was cultured under illumination immediately after induction, produced tocopherols by 1.48 times as much as those in Ca-2, even in a static culture.By this research, it was clearly shown that the formation of "green cells, which were contained abundantly chloroplasts, is a prerequisite for the increase of tocopherols. Therefore, we now investigate the establishment of high producing strain by greening, i.e. the formation of chloroplast. Less