缺氧胁迫小细胞团法选育高产黄酮苷悬浮细胞系

Research on the small cell groups with hypoxia stress to select suspension cell line of higher productivity of flavonol glycoside

以银杏优良品种的种苗诱导愈伤组织 ,考察了B5,SH ,MS ,White 4种培养基对银杏愈伤组织生长性状和黄酮苷合成能力的影响 ,采用缺氧胁迫小细胞团法从愈伤组织中筛选高产黄酮苷的细胞系 ,并对细胞系在悬浮培养时的条件进行了优化 .结果表明 :TZG 1培养周期 18d ,生长指数为 4.12 ,黄酮苷含量为 1.2 5 % ,比愈伤组织提高了2 5 7.1% ;MS培养基有利于形成质地松软的愈伤组织 ,当培养基中NH+4 与NO-3 的浓度比为 2 0 .6 / 39.4或 10 .3/ 2 9.1,蔗糖质量浓度为 30g/L或 40g/L ,pH值为 5 .6～ 6 .0时 ,有利于细胞生长和黄酮苷合成 .

The effects of media (B_5, SH, white, MS) on the growth and synthesis of flavonol glycoside of calli introduced from seedling of Ginkgo biloba, the cell lines for suspension culture selected by hypoxia stress for small cell groups, and the factors influencing cell growth and synthesis of flavonol glycoside in suspension culture were studied. The results show that MS media are suitable to introducing callus of Ginkgo biloba L, cell line selected has 4.12 of growth index and 1.25%(dry mass fraction of cells) of flavonol glycoside that is enhanced by 257.1% compared with calli, and the c(NH+_4)/c(NO-_3)(20.6/39.4 or 10.3/291), sucrose content (30～40 g/L), pH(5.6～6.0) are beneficial to cell growth and synthesis of flavonol glycoside.