Panax ginseng C. A. Meyer is a desirable medicinal plant due to its health benefits. With the growing interest in ginseng products, the safe preservation of genetically engineered material with unique attributes is increasingly important. In the present study, an effective protocol for cryopreservation of the ginseng embryogenic callus by vitrification is reported. After 2-wk subculture, the calluses were cold-acclimated at 4°C for 3 wk, then precultured for 2 d with a preculture medium containing 0.3 M sucrose. The precultured embryonic callus was then treated with a loading solution, followed by exposure to plant vitrification solution 2 at 0°C for 90 min. The dehydrated embryogenic calluses were then transferred into fresh 1.5 mL plant vitrification solution 2 and directly immersed in liquid nitrogen. With this protocol, an average of 68.3% regrowth rate for cryopreserved material was obtained, and the number of somatic embryos differentiated from embryogenic callus was significantly increased. Proteomic analysis showed that proteins related to carbohydrate metabolism, the stress response, and oxidative metabolism all significantly up-regulated after cryopreservation. Our results indicate that cold acclimation and preculture may be linked to enhanced cryopreservation tolerance. The up-regulated proteins play an important role in ginseng stress responses and may contribute to cryopreservation tolerance. Carbohydrate metabolism–related protein may be associated with the recovery growth of the cryopreserved embryogenic callus. The increased expression of HSP and 14-3-3-like protein play important roles in the response to stress and could be related with the improvement of somatic embryo production after cryopreservation.

人参由于其对健康的好处，CA Meyer是理想的药用植物。随着人参产品的兴趣日益浓厚，具有独特属性的基因工程材料的安全保存变得越来越重要。在本研究中，报道了通过玻璃化冷冻保存人参胚性愈伤组织的有效方案。2周传代培养后，将愈伤组织在4°C下冷驯化3周，然后用含有0.3 M蔗糖的预培养基预培养2天。然后用加载溶液处理预培养的胚愈伤组织，然后在0℃下暴露于植物玻璃化溶液2 90分钟。然后将脱水的胚性愈伤组织转移至新鲜的1.5 mL植物玻璃化溶液2中，并直接浸入液氮中。使用此协议，平均为68​​。低温保存的材料的再生长率为3％，从胚性愈伤组织分化出来的体细胞胚的数量显着增加。蛋白质组学分析表明，与糖代谢，应激反应和氧化代谢相关的蛋白质在冷冻保存后均显着上调。我们的结果表明，冷驯化和预培养可能与提高冷冻保存耐受性有关。上调的蛋白质在人参应激反应中起重要作用，并可能有助于冷冻保存的耐受性。碳水化合物代谢相关蛋白可能与冷冻保存的胚性愈伤组织的恢复生长有关。