玻璃化法及小液滴玻璃化法对香石竹茎尖超低温保存效果的影响

Effects on Cryopreservation by Vitrification and Droplet-Vitrification of Dianthus caryophyllus L.

[目的]探索更有效的香石竹茎尖超低温保存方法,为球宿根花卉的超低温保存提供相关技术支持。[方法]以香石竹茎尖为材料,研究预培养条件(蔗糖浓度、预培养时间)、玻璃化处理时间和玻璃化方法(常规玻璃化法、小液滴玻璃化法)对香石竹茎尖超低温保存效果的影响。[结果]在0.5 mol/L的蔗糖浓度培养基上预培养5 d香石竹茎尖成活率最高,为88.3%;玻璃化处理80 min成活率最高,可达86.7%;常规玻璃化法成活率及再生率在80%以上,小液滴玻璃化法成活率可达90%以上且全部再生,小液滴玻璃化法再生天数比常规玻璃化法少3~5 d。[结论]小液滴玻璃化法是一种有发展前景的超低温保存方法。

[Objective] To explore a more effective technique for Dianthus caryophyllus L. shoot-tips cryopreservation,and to provide technical basis for the bulbs flowers cryopreservation. [Method]With D. caryophyllus shoot-tips as the materials,the effects of pre-culture treatments（ sugar concentrations,pre-culture time）,vitrification time and vitrification methods on survival rate,regeneration rate and regeneration time of cryopreservation were studied. [Result] When pre-cultured in the medium with 0. 5 mol/L sucrose for 5 d,the survival rate was 88. 3%.The survival rate was up to 86. 7% under the vitrification treatment for 80 min. The survival rate and regeneration rate of conventional vitrification method were above 80%,the survival rate of droplet-vitrification method could reach more than 90% and regeneration rate was 100%.The number of regeneration days of droplet-vitrification was 3-5 d less than conventional vitrification. [Conclusion] Droplet-vitrification is a promising method for bulbs shoot-tips cryopreservation.