Effects of Nutritional Conditions on Tobacco (Nicotianatcbfeum L) Cell Suspension Culture

담배세포 (Nicotiana tabacum) 의 액체배양에 관한 연구

For the preliminary experiments of mass Production of tobacco cells in tank culture, the effects of nutritional conditions on the growth of suspended cells were investigated ; 1. The tobacco cell growth was affected by concentrations of sucrose or inorganic phosphate, type of nitrogen source, and plant hormone, especially 2, 4-D. 2. The optimum level of sucrose concentration was 3% and the level of inorganic phosphate was 0.3mg /ml, which was about twice as high as the level of Linsmaier - Skoog medium. 3. The best growth was observed when the ratio of nitrate nitrogen to ammonium nitrogen was 2 : 1, where the total nitrogen content was equal to that of nitrogen source. 4. To find out the mechanism of promotive effects of 214-D and inorganic phosphate on the tobacco cell growth, the respiration and metabolism of $^{14}\textrm{C}$-91ucose were investigated. Addition of 2, 4 -D in culture medium increased if 2, 4-D (0.2ppm )was added to medium or the level of inorganic Phosphate was raised 2.5 times as high as standard. In cultures with high inorganic phosphate and 2, 4-D, the absorbed 14C-glucose was converted to amino acids and organic acids rather than remained as sugars.

담배 (Va 115) 細胞의 液 培養에서 담배細胞의 多量生産을 할 수 있는 Tank 培養에 關한 基礎調査로 培地造成의 果를 調査하였으며 細胞增殖率에 큰 影響을 미쳤던 2,4-D와 無機燐酸의 果에 關하여 調査하였다. 1. 培地造成에서 細胞增殖率에 많은 影響을 미쳤던 要素는 무糖, 無機燐酸의 濃度,窒素源의 形態 및 植物홀몬,特히 2, 4-D의 有無였다. 2. 무糖의 最適濃度는 3%였으며 3%以上의 濃度에서는 多少 細胞增殖率이 좋은 듯 하였으나 그 差異는 크지 않아 3% 程度로 足하였다. 無機燐酸濃度는 LS 培地內의 無機燐酸의 約 2.5 培인 0.30mg/ml 일 때 細胞增殖率이 가장 좋았다. 3. 液 培地의 窒素源은 암모니아態窒素와 硝酸態窒素가 1 : 2 일 때가 가장 좋았고 窒素源이 암모니아태 만으로 使用하였을 때 細胞增殖率이 가장 낮았으며 硝酸態만 使用되었을 때는 암모니아태만 쓰였을 때 보다는 좋았으나 암모니아태와 硝酸態의 比가 1 : 2 일 때 보다는 떨어졌다. 4. 液 培地에 2,4-D 添加와 無機燐酸濃度를 높이면 細胞增殖率이 增加되는 機作을 調査하기 爲하여 呼吸率과 14C - glucose 吸收利用을 調査하였다. 細胞의 吸收率은 2, 4-D를 添加하면 增加되며 14C-glucose의 吸收는 培地內에 2, 4-D가 包含되거나 (0.2 ppm) 燐酸濃度가 높아지면(對照의 2.5培) 더욱 많았고, 吸收된 14C-glucose는 糖 상태보다 다른 形態,特히 amino 酸이나 有機酸으로 많이 变하였다.