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

通过使用不同的生长调节剂和培养基添加剂进行筛选，获得了生长速度和生育酚含量均较高的Caf-B2KC细胞系。此外，生育酚的生产受到生物合成前体的刺激，特别是生育酚侧链的前体植醇。生育酚的总含量增加了约5倍。此外，从光照下的幼苗中分离到一株淡绿色菌株Ca-2，其总生育酚的产量是Caf-B2KC的1.4倍。然而，Ca-2菌株在任何处理下都没有进一步变深，生育酚含量也没有增加。本研究的主要目的是通过以下3种方法通过绿化提高红花培养细胞中生育酚的含量。(1)利用…技术，将CaF-B2KC愈伤组织与原植物叶肉细胞通过每一个原生质体进行电细胞融合更多的电子细胞融合设备。(2)用电穿孔法将叶肉叶绿体导入Caf-B2KC愈伤组织；(3)用微量注射法将叶绿体注入Caf-B2KC愈伤组织。培养细胞和叶肉细胞均能高产量地形成原生质体，但对渗透压和其他冲击等物理变化非常不稳定。因此，用上述三种方法都不能形成含有大量叶绿体的暗绿色愈伤组织，即使从原始植物的叶肉中分离出原生质体。一株绿色新菌株Ca-3在诱导后立即在光照下培养，即使在静态培养中，其产生的生育酚也是Ca-2的1.48倍。本研究清楚地表明，含有丰富叶绿体的绿色细胞的形成是增加生育酚的先决条件。因此，我们现在研究通过绿化建立高产菌株，即形成较少的叶绿体。

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