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以生姜的脫病毒試管苗在繁殖培養(yǎng)基上培養(yǎng)25d獲得的叢生芽為試材,探討生姜超低溫玻璃化法保存的最優(yōu)程序。結(jié)果表明:采用繼代培養(yǎng)45d的叢生芽,切取2~3mm,于0.5mol/L蔗糖濃度的MS培養(yǎng)基內(nèi)預(yù)培養(yǎng)2d,60%PVS2室溫處理5min,100%PVS2 0℃下處理30min,迅速投入液氮,48h后,37~40℃水浴快速化凍2min,1.2mol/L蔗糖的MS培養(yǎng)液洗滌20min,在MS+0.5mg/L BA+0.1mg/L NAA+0.3mg/L GA3+30g/L蔗糖+7g/L瓊脂的培養(yǎng)基上恢復(fù)培養(yǎng)的成活率最高,為57.7%。
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A procedure on cryopreservation of shoot-tips of Eriobotrya japonica Lindl.had been studied in vitro by two steps vitrification.The effects of cold-hardening,preculture,cryoprotecten treatment,thawing and reculture on survival rate of the shoot-tips were tested.Optimal media for shoot-tip induction and propagation were 1/2 MS+1.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1)NAA and MS+0.5 mg·L~(-1)6-BA+0.1 mg·L~(-1) NAA.Preculture for 30 min with 60% PVS_(2)(30% glycerol+15% ethylene glycol+15% DMSO+0.4 mol·L~(-1) sucrose) and treatment for 50 min by PVS_(2) were suitable to shoot-tip cryopreservation.After cryopreservationsome shoot-tips couldregenerate plantlets,the regenerate rate was 46.8%.There was no difference in regenerating plantlets and tube plantlets not stored by cryopreservation.
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