目的 选育高产银杏内酯悬浮细胞系，并对其在继代培养过程中银杏内酯B合成的稳定性进行研究。方法以银杏优良品种的种子萌发幼苗诱导愈伤组织，考察了不同培养基对愈伤组织生长和银杏内酯B产生的影响，采用缺氧胁迫法从诱导的愈伤组织中选育高产银杏内酯B悬浮细胞系，并进行驯化培养。结果 选出了7个细胞系，其合成银杏内酯B的能力比选育前的愈伤组织有了显著提高，其中细胞系MH－3培养周期18d，细胞生物量增加3．71倍，银杏内酯B含量达到302μg/gDW，比选育前提高了172．7%，为国内领先水平。在摇瓶培养时其合成银杏内酯的能力稳定，连续培养6代，银杏内酯B的平均含量为291μg/gDW，变异系数为0．131。结论 采用该路线选育的银杏悬浮细胞系银杏内酯B产量高，在继代培养过程中合成代谢较稳定。

Object To select the cell line of Ginkgo biloba L. that may produce high yield of ginkgolide B (GB) in suspension culture and to study the stability of GB in subculture. Methods Calculus was induced with stem, root and leaf of selected elite species and high yield suspension cell lines chosen by hypoxia stress. Results Seven suspension cell lines with improved yield of GB were obtained. Among which cell line M H-3 gave a 3.71 fold increase of cellular bioproduct with an increased content up to 302μg/gDW after culturing for 18d, a leading record country wide. In shaking bottle culture, it showed a consistent yield of GB with a content of 291μg /g DW in 6 successive subcultures, coefficient of variation=0.131. Conclusion The suspension cell line selected by hypoxia stress gave a higher yield and stability in successive transfer culture.

广东省自然科学基金(No．990865)；中南大学科研基金资助项目