关键词: 野葛, 茎尖, 玻璃化法, 超低温保存, 再生率

Abstract: The cryopreservation technique of Pueraria lobata shoot-tip was studied in this paper.The results showed that the better procedure of cryopreservation by vitrification was as follows.The aseptic plantlets of H^# Pueraria lobata subcultured in vitro for 30 days were treated at 4℃ for 5 days.The shoot-tips 1.5~2.5 mm in length with one or two leaf primordium of H^# Pueraria lobata were cut and precultured at 4℃ for 1 day in MS supplemented with 5% DMSO+5% sucrose.They were dehydrated with 60% and 100% PVS₂(PVS₂:30% glycerol+15% glycol+15% dimethyl sulfoxide+13.7% sucrose) at 0℃ for 30 minutes, respectively.The shoot-tips were immersed immediately into liquid nitrogen directly and conserved for 24 hours.After rapidly thawing in a water bath at 40℃ for 90 seconds, the shoot-tips were washed two times with MS supplemented with 41.1% sucrose (10 minutes each time), then transferred and re-cultured on the MS medium supplemented with BA 2 mg/L and NAA 1mg/L in dark for 7 days and then in light.The survival rate was up to 60%.The regenerated plantlets of Pueraria lobat were the same as the normal plantlets in morphology.

Key words: Pueraria lobata, shoot-tip, vitrification, cryopreservation, regeneration rate