2024年10月31日 星期四
凹叶厚朴愈伤组织的超低温保存
Calli cryopreservation of Magnolia biloba (Rehd et Wils) Cheng
1996年 第5卷 第1期 页码[9-13]    下载全文[0.6MB]  
摘要

采用单因子比较和均匀设计法,研究凹叶厚朴(Magnolia biloba(RehdetWils)Cheng)愈伤组织超低温(-196℃)保存的影响因素。结果表明:超低温保存的适宜材料是4周龄的愈伤组织。经4~6℃低温下预培养12天的愈伤组织抗冻能力最强。较好的冷冻保护剂是12.5%DMSO+5%Suc+0.25%LH。较佳的冷冻程序是以0.5℃/min的降温速度从0℃降到-30℃,停留1h,然后浸入液氮中贮存。30℃水浴中化冻与自来水流水(18.5±1℃)冲洗化冻效果一样良好。化冻后的愈伤组织在黑暗中培养生长较好,存活率高。

Abstract

The suitable factors in cryopreservation ( - 196℃) of the calli of Magnolia biloba  (Rehd et Wils) Cheng were studied by monofactorial and uniform. design. The experiments showed that four-weeked calli were the best material for cryopreservation. Calli treated for 12 days at 4 - 6℃were the best cryotolerant. The best cryoprective was the mixture of 12.5 % DMSO, 5 % Suc and 0.25 % LH. The best freezing procedure was to reduce the temperature from O℃ to - 32℃ at a stable speed by lowerring 0.5 each minute , leave the calli at - 3 2℃ for 1 h , then drop them in liquid nitrogen
( - 196℃ ) to be preserved. Warm-water bath at 30 was as good as flowing-water wash at 18.5± 1℃for melting the frozen material. Unfrozen calli were then cultured in darkness and grew well with a high survival rate.

关键词凹叶厚朴; 超低温保存; 愈伤组织;
Key wordsMagnolia biloba (Rehd et Wils) Cheng; cryopreservation; calli
作者刘贤旺,杜勤
所在单位江西中医学院,南昌330006
点击量8258
下载次数985
基金项目国家自然科学基金;