Research Center for Cell and Tissue Culture, Kyoto University, Sakyo-ku, 606, Kyoto, Japan.
Plant Cell Rep. 1986 Feb;5(1):50-3. doi: 10.1007/BF00269717.
Cultured R. serpentina cells have been maintained on modified Linsmaier-Skoog medium for over 13 years. These cultured cells produced much more ajmaline (0.005-0.012% dW) than reserpine (0-0.003% dW). Selection of callus which survived the stress induced by alteration of the medium composition including hormones, was repeated over several generations. Surviving callus was then transferred back to the original liquid growth medium and subculture continued, during which time the cells exhibited a return to their pre- stress rate of growth, enhanced reserpine production, and a decrease in ajmaline production. R. serpentina cell suspension cultures selected as described and serially subcultured in fresh growth medium every 3 weeks consistently produce reserpine at a yield of approximately 0.03-0.06% dW.
经过 13 年的时间,培养的 R. serpentina 细胞一直保存在改良的 Linsmaier-Skoog 培养基上。这些培养的细胞产生的阿马林(0.005-0.012% dw)比利血平(0-0.003% dw)多得多。通过选择在包括激素在内的培养基成分改变所诱导的应激下存活下来的愈伤组织,在几代中重复进行选择。然后将存活的愈伤组织转回原始的液体生长培养基中,并继续进行继代培养,在此期间,细胞恢复到应激前的生长速度,利血平的产量增加,阿马林的产量减少。如所述选择的 R. serpentina 细胞悬浮培养物,并在新鲜生长培养基中每 3 周进行连续继代培养,始终以约 0.03-0.06% dw 的产率产生利血平。