美丽崖豆藤细胞培养及其多糖质量分数分析

Cell culture of Millettia speciosa Champ. and its polysaccharide content analysis

采用不同激素诱导美丽崖豆藤(Millettia speciosa Champ.)无菌苗下胚轴、叶片和茎尖产生愈伤组织,对适宜的愈伤组织进行了液体培养,并对培养物与美丽崖豆藤干燥根(牛大力)切片的多糖质量分数进行了比较.结果表明:美丽崖豆藤无菌苗下胚轴、叶片和茎尖在不同的培养基上均形成愈伤组织,无器官分化发生;下胚轴在含1 mg/L 2,4-D培养基上形成的乳白色、易碎愈伤组织在液体培养基MS+1mg/L 2,4-D+0.75 g/L酵母提取物+0.75 g/L水解酪蛋白中增殖速度很快,能以悬浮细胞形式增殖9代,之后培养物中开始出现较大的愈伤组织块;悬浮细胞及后期形成的愈伤组织团块多糖质量分数分别为2.01%和2.07%,高于牛大力切片多糖的1.04%,且差异极显著(P<0.01).表明美丽崖豆藤细胞培养可作为提取美丽崖豆藤多糖的重要途径.

Excised hypocotyl segments, leaves and shoot tips from sterile seedlings of Millettia speciosa were used to induce callus in MS solid medium with various phytohormones and the suitable callus was further cultured in MS liquid medium. Polysaccharide contents in different cultures and roots of M. speciosa were detected and compared by phenol-sulfuric acid method. The results showed that all explants （hypocotyl, leaves and shoot tips） formed callus without organogenesis in MS solid medium containing 1 mg/L 6 - Benzylaminopurine （ 6 - BA）, 1 mg/L Thidiazauron （TDZ） or 1 mg/L 2,4 - dichlorophenoxyacetic acid （2,4 -D） ; Excised hypocotyl segments cultured on MS medium containing 1 mg/L 2,4 - D produced creamy and fragile callus, which grew fast when transferred into MS liquid medium supplemented with 0. 75 g/L yeast extract, 0. 75 g/L acid hydrolyzed casein, 1 mg/L 2,4 - D. This callus dis- persed well and kept for 9th subcuhure. However, after 9th subculture, some large cell aggregates began to form. Well dispersed callus and large callus clumps contained 2. 01% and 2. 07% polysaccharides, respectively, more than that in roots of M. speciosa, which indicated that cell culture was a significant way to which was 1.04% Millettia speciosa polysaccharides.