青阳参组织培养及愈伤组织的成分分析

用青阳参（Cynanchum otophyllum）的嫩枝和芽在MS + 2.0 mg/L 2,4-D + 0.1 mg/L KIN的培养基上诱导愈伤组织。通过不同的培养基和激素配比实验，发现 6,7-V + 2.0 mg/L 2,4-D + 0.3 mg/L KIN 最适合愈伤组织的生长。但在6,7-V + 1.0 mg/L 2,4-D + 0.1 mg/L KIN 培养基中的愈伤组织次生代谢物含量最高。愈伤组织的生长周期为27 d，但在33 d时次生代谢产物的含量最高。从愈伤组织中分离到7个化合物：⑴9,10,11-三羟基-十八碳-12(Z)-烯酸甲酯 (methyl 9,10,11-trihydroxy-12-octadecenoate)，(2) 胡萝卜甙 (daucosterol)，(3)β 谷甾醇 (β sitosterol)，（4） 华木酸 (betulinic acid)，（5）齐端果酸 (oleanolic acid)，（6）棕榈酸 (hexadecanoic acid)，（7）十八碳-9-烯酸 (9-octadecenoic acid)。首次报道从植物愈伤组织中分离到多羟基十八碳烯酸，并讨论了化合物（1）对植物细胞生长的可能影响。

Studies on the Tissue Culture of Cynanchum otophyllum and Calli Chemical Constituents

The calli of Cynanchum otophyllum were induced from the tender stems and young buds on MS medium with the supplement of 2.0 mg/L 2,4-D and 0.1 mg/L KIN. The result showed that 6,7-V medium was the optimal medium for callus growth. The tests using plant growth regulators with various concentrations showed that 6,7-V medium plus 2.0 mg/L 2,4-D and 0.3 mg/L KIN was the best combination for callus growth, whereas the production of secondary metabolites was higher on 6,7-V medium plus 1.0 mg/L 2,4-D and 0.1 mg/L KIN. The best period of culture was 27 d for callus growth, and 33 d for the maximal production of secondary metabolites. The isolation of nature products from the callus provided seven compounds including (1) fatty acid methyl ester named methyl 9,10,11-trihydroxy-12-octadecenoate and (2) daucosterol, (3) β sitosterol, (4) betulinic acid, (5) oleanolic acid, (6) hexadecanoic acid, (7) 9-octadecenoic acid. This is the first report of the isolation of poly-hydroxyl octadecenoic acid from a callus, and its proven biological function on the growth of plant cells was discussed.