银杏悬浮培养细胞的生长、分化与萜内酯化合物的积累

Studies on the Cell Growth, Differentiation and Terpene Lactone Accumulation in Ginkgo biloba Cell Suspension Cultures

研究了来源于银杏种子胚和幼苗茎的悬浮细胞的生长、分化和培养物中的白果内酯、银杏内酯A和B的含量变化。结果表明 :在悬浮培养中 ,细胞聚集而成的细胞团大小、细胞中叶绿体的分化、外植体来源都影响培养物中的萜内酯的种类和含量 ,胚来源的悬浮细胞培养物中 ,银杏内酯B仅存在于直径 <2mm的小细胞团悬浮培养中 ,且在 <1mm的细胞团中的含量最高 ,达 0 4 37mg g(DW) ;而直径 >3mm的细胞团悬浮培养物中只含有白果内酯和银杏内酯A。相同大小的悬浮细胞团中 ,胚来源的细胞中萜内酯含量高于茎来源的细胞。

To provide supports for Ginkgo biloba cell engineering for production of Terpene lactones(Ginkgolides and bilobalide),the cell suspension were established from calli induced from zygote embryos and stems of 30-day-old seedlings respectively. The relationship between cell growth, differentiation and the terpene lactone accumulation in these suspension cultures were investigated. HPLC determination indicated that,the ginkgolide B was found in the embryo derived cell suspension cultures at 0.044% of cell dry weight, and this result was the first time reported in this study. The accumulation of terpene lactone in the suspension cultures derived from both the embryo and seedling stems are effected by the level of the cell differentiation. The ginkgolide B was only found in small cell aggregates in the size smaller than 2mm, and the highest level of ginkgolide B was accumulated in cell aggregates in the size smaller than 1mm; however, the cell aggregates in the size bigger than 3mm could only produced bilobalide and ginkgolide A. In the same size aggregates of the suspension cultures the terpene lactone accumulation is strongly effected by the source of the explant. When the size of cell aggregates was in less than 1mm, the concentration of bilobalide, ginkgolide A and B in the cell suspension cultures derived from the embryos was 2, 1.4 and 0.56-fold, respectively, higher than that of cell cultures derived from seedling stems.