提高银杏悬浮培养细胞内酯合成的研究

Studies on Increasing Ginkgolides Synthesized in Suspension Cell Culture of Ginkgo biloba L

对 12个不同品种的银杏树幼叶及其诱导的愈伤组织进行银杏内酯含量测定 ,发现不同基因型的银杏内酯含量存在较大差异 :圆铃和金坠两个品种含量最高 ,分别达 0 .43%和 0 .41%;七星果品种含量最低 ,为 0 .13%;金坠品种幼叶愈伤组织中银杏内酯含量最高 ,为 0 .0 334%;有 3个品种的幼叶愈伤组织未检测到银杏内酯。选取银杏内酯含量高、生长旺盛的愈伤组织在B5液体培养基中进行了细胞悬浮培养 ,研究多种生长因子对细胞生长和银杏内酯合成的影响。结果表明 ,在 15 0mL培养瓶中装 5 0mL培养液 ,接种量为 30～ 40g/L ,培养液初始pH为 5 .8,3 0 0 0～ 40 0 0lx的强光照 ,以 30g/L蔗糖和 15g/L葡萄糖为碳源最有利于细胞生长和银杏内酯的合成。用高效液相色谱检测结果显示 ,银杏圆铃品种的悬浮培养细胞中银杏内酯B最高含量可达细胞干样质量的 0 .0 75 8%。

After determination of the content of ginkgolides from varieties of Ginkgo biloba L.,it was shown that there were significantly different contents of ginkgolides from different genotypes of Ginkgo biloba L.,in which the highest were in Yuanling and Jinzhui,the contents reached 0.43% and 0.41% respectively,the lowest was in Qixingguo,the content was 0.13%,the highest content of ginkgolides in calli from varieties of Ginkgo biloba L.was in Jinzhui and reached 0.334‰.There were calli from 3 varieties producing no ginkgolide.The callus with high content of ginkgolides and growing well was cultured on B 5 media.The effects of different growth factors on growth of cell and content of ginkgolides were studied.The result showed that 50mL culture solution in the 150mL flask,the inoculation 30-40g/L,the initial pH in culture solution 5.8,3000-4000lx lumination in the intensity and the mixture of 30g/L sucrose and 15g/L glucose were optimum to synthesis of ginkgolides and growth of cell.The result by HPLC detection showed ginkgolides in suspension cells reached 0.0785% of dry mass of cells from the variety Yuanling.