Study on algal growth factor in agar and production of bioactive substance by tissue culture

琼脂中藻类生长因子的研究及组织培养生产生物活性物质

• 批准号: 03660209
• 负责人: AMANO Hideomi
• 金额: $ 1.22万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03660209/
• 关键词: Seaweed; Tissue culture; Growth factor; Agar; Bioactive substance; Antitumor substance; Physicochemical property; マツモ; コブシミル; ミル; 有機酸

1. Identification of algal growth factor in agar. Gracilaria agar has an unknown algal growth factor having low molecular weight. Using reversed-phase (Shimadzu Shim-pack CLC-ODS) and silica gel (TOSO TSK gel Amido-80) HPLC, X-ray fluorescence analysis and atomic absorption spectrometry, sugars, organic acids, amino acids, nucleic acid-related compounds, inorganic elements were analyzed. Gracilaria agar contained tartaric, malonic and citric acids which are not detected in Gelidium agar. When those three acids were added to culture medium, growth rate of green alga Ulva pertusa increased to 149% of that of control. Those three organic acids, there for, were identified as the growth factor in Gracilaria agar. 2. Physicochemical properties of the agar for algal tissue culture. The 0.7% Gracilaria agar showed the best growth of algal tissue. The gel strength, pH, viscosity, SO_4 contents, syneresis after 200 h of the 0.7% agar were 24, 6.6, 7.9 cp, 0.79%, 2 g, respectively. The most suitable agar for algal tissue culture is considered to have moderate gel strength and small syneresis. 3. Production of antitumor substance by tissue culture. Axenic tissue from brown alga Heterochordaria abietina, green algae Codium pugniformis, and C. fragile were cultured in PESI medium at 15 C on a 14:10 L:D cycle under 1200 lx. Yield of antitumor glycoprotein was 4.7%, 2.1% and 2.2% in H. abietina, C. pugniformis and C. fragile, respectively. Inhibitory effects of the glycoprotein on growth of WI-38 VA-13 were examined. IC_<50> was 9.1, 300.8 and 274.9 in H. abietina, C. pugniformis and C. fragile, respectively. These values are almost the same as that of naturally grown algae in ocean. By the axenic tissue culture, therefore, antitumor glycoprotein was produced successfully.

1.琼脂中藻类生长因子的鉴定。江蓠琼脂含有一种未知的低分子量藻类生长因子。采用反相（Shimadzu Shim-pack CLC-ODS）和硅胶（TOSO TSK gel Amido-80）高效液相色谱法、X射线荧光分析法和原子吸收光谱法，对糖、有机酸、氨基酸、核酸相关化合物、无机元素进行了分析。江蓠琼脂含有石花菜琼脂中未检出的酒石酸、丙二酸和柠檬酸。在培养基中添加这三种酸，绿色孔石莼的生长速度提高到对照的149%。因此，这三种有机酸被鉴定为江蓠琼脂中的生长因子。2.藻类组织培养用琼脂的理化特性。0.7%江蓠琼脂对藻组织生长的影响最大。0.7%琼脂的凝胶强度为24，pH值为6.6，粘度为7.9 cp，SO_4含量为0.79%，200 h后的脱水率为2 g。认为适宜藻类组织培养的琼脂应具有中等的凝胶强度和较小的脱水收缩。3.通过组织培养生产抗肿瘤物质。本文报道了从棕树异索藻（Heterochordaria abietina）、绿色海藻松藻（Codium pugniformis）和松藻（C.在PESI培养基中于15 ℃、1200 lx下以14：10 L：D循环培养脆性细胞。抗肿瘤糖蛋白在H.冷杉C. pugniformis和C.脆弱，分别。检测了糖蛋白对WI-38 VA-13生长的抑制作用。对<50>H的IC_2分别为9.1、300.8和274.9。冷杉C. pugniformis和C.脆弱，分别。这些值与海洋中自然生长的藻类几乎相同。因此，通过无菌组织培养，成功地生产了抗肿瘤糖蛋白。

项目成果

Hideomi Amano: "Effect of plant growth regulators, organic acids, and sugars on growth and chemical compositions of tissue from sea lettuce Ulva pertusa" Nippon Suisan Gakkaishi. 59. (1993)

Hideomi Amano：“植物生长调节剂、有机酸和糖对海生菜 Ulva pertusa 组织生长和化学成分的影响”Nippon Suisan Gakkaishi。

H.Amano: "Effect of plant growth regulators,organic acids,and sugars on growth and chemical compositions of tissue from sea lettuce Ulva pertusa" 日本水産学会誌. 59. (1993)

H.Amano：“植物生长调节剂、有机酸和糖对海生菜 Ulva pertusa 组织的生长和化学成分的影响”，日本水产协会杂志 59。（1993 年）。

天野 秀臣: "海藻組織培養における生長促進物質に関する研究" 日本水産学会誌.

天野英臣：《海藻组织培养中生长促进物质的研究》日本水产学会会刊。

H.AMANO: "Effect of plant growth regulators,organic acids,and sugars on growth and chemical compositions of tissue from sea lettuce Ulva porusa" 日本水産学会誌. 59. (1993)

H.AMANO：“植物生长调节剂、有机酸和糖对海生菜 Ulva porusa 组织的生长和化学成分的影响”，日本水产科学学会杂志 59。（1993 年）。

M.Polne-Fuller: "Enhanced development and differentation of protoplasts and spores of green and red seaweeds by a Pterocladia agar from New Zealand" Hydrobiologia. (1993)

M.Polne-Fuller：“来自新西兰的 Pterocladia 琼脂增强了绿色和红色海藻原生质体和孢子的发育和分化”Hydrobiologia。