华重楼超低温保存条件的优化研究  

Optimization of Cryopreservation Condition for Paris polyphylla var. chinensis

作　　者：王跃华[1] 刘银花[1,2] 陈燕 彭世明 唐旭[1,2] 何新友 任鹏飞 何诗红 

机构地区：[1]成都大学生物产业学院,四川成都610106 [2]四川师范大学生命科学学院,成都610101 [3]四川金土地中药材种植集团有限公司,成都611130

出　　处：《种子》2015年第4期13-16,共4页Seed

基　　金：四川省科技支撑计划项目(编号:2014 ZC 2103);成都市科技局项目(2014)

摘　　要：目的:获得华重楼地下茎超低温保存的关键技术。方法:选取预培养基中二甲基亚砜的浓度、保存材料的预培养时间、装载时间和脱水时间为正交设计的4因素,每个因素确立4个水平,并以细胞相对存活率为考察指标,对华重楼地下茎的超低温保存条件进行优化研究。结果:华重楼地下茎的最佳保存条件是在含5%二甲基亚砜的液体培养基中预培养5 d后,用60%玻璃化溶液装载45 min,再用100%玻璃化溶液在4℃条件下脱水处理60 min,其保存后细胞的相对存活率可高达92.10%。结论:华重楼地下茎种质资源的玻璃化超低温保存操作简单、成活率高,可广泛地用于生产实践。Objective： To establish the critical techniques for cryopreservation of Paris polyphylla var. chinensis.Method： The concentration of dimethyl sulfoxide,pre-cultivation time,loading time,and dehydration time were optimized in an orthogonal design. Four different levels were tested for each of the factor. The results were evaluated by the relative survival rate of the cells. Result： The optimal cryopreservation condition for Paris polyphylla var. chinensis was pre-cultivation in 5% DMSO for 5 days,followed by 45 minutes loading with 60%vitrified solution. The sample was then dehydrated with 100% vitrified solution for 1 hour. Under such condition,the relative survival rate was 92. 10%. Conclusion： The method of vitrification to cryopreserve the germplasm of P. polyphylla var. chinensisis simple in handling with high in survival and can be applied in practice.

关 键 词：华重楼 超低温保存 正交设计 

分 类 号：S567.239[农业科学—中草药栽培]