我国目前处于濒危状态的药用植物近2100种，保护难度极大。玻璃化超低温保存是永久保存种质的理想方法，但植物种类、组织器官或生理状态不同，保存的具体方法也各不相同，材料冻存后再生率低。超低温保存机制不清已成为其技术发展的主要瓶颈。大量研究表明，组织器官类型及其生理状态是玻璃化超低温保存的重要内在因素，与材料的耐冻、耐脱水能力密切相关。休眠芽不仅抗逆性强，超低温保存后再生率高，还具有很强的形态发生能力。前期研究发现，低温锻炼时间是党参休眠芽超低温保存的关键因素。本项目首次以完整休眠芽作为玻璃化超低温保存材料，通过研究不同低温锻炼时间的休眠芽在超低温保存逆境中生理指标、蛋白质组、细胞超微结构和组织化学的差异，明确各指标变化与逆境响应的关系，为通过材料的选择及生理状态的调控提高保存效果提供理论支撑。进一步研究超低温保存逆境对再生植株形态、分子水平变异和农艺性状产生的影响，评价该技术方法的实用性。

At present, there are about 2100 species of medical plants in imminent danger and their protection is very hard. Cryopreservation is an ideal way to permanently preserve germplasm. However, different plants, organs and physiological state, procedure of cryopreservation are different. Many cryopreserved material still had low regeneration. Having little knowledge of mechanism of cryopreservation is the bottleneck of the technology. A lot of researches showed type of organs and physiological state were the important internal factors which relate to the tolerance to dehydration and freeze. Studies showed that dormant buds have great morphogenesis ability and strong stress resistance and high rate of cryopreservation survival. Preliminary study showed that cold acclimation is the key factor in cryopreservation of whole dormant bud of Codonopsis pilosula (Franch.) Nannf. In this project, a comparative research on antioxidant system, osmotic adjustment ability, proteome, histochemical localization and cell microstructure of dormant buds of different acclimation time will be studied in the process of vitrification cryopreservation to exploit the change of the parameters and their relationship with stress response. The purpose of this work is to provide theoretical guidance for improving the survival rate of plant material after cryopreservation by choosing suitable material and adjusting it to fitting physiological state. Moreover, phenotype, agronomic trait and moleculer genetics of regenerated plants will be studied, which can provide an objective evaluation of practicality of the technology.