提高银杏悬浮培养细胞内酯合成的研究

 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. 0334%; 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150 mL 培养瓶中装50 mL 培养液, 接种量为30~ 40 g/ L, 培养液初始pH 为5. 8, 3 000~ 4 000 lx 的强光照, 以30 g / L 蔗糖和15 g/ 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. 0758%。

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

After determination of the content of ginkgolides from varieties of Ginkgo biloba L. , it was shown that therewere 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 variet ies 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 B5 media. The effects of different growth factors on growth of cell and content of ginkgolides were studied. The result showed that 50 mL culture solution in the 150 mL flask, the inoculat ion 30- 40 g/L, the initial pH in culture solution 5. 8, 3 000- 4 000 lx lumination in theintensity and the mixture of 30 g/ L sucrose and 15 g/L glucosewere 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 cellsfrom the variety Yuanling.