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园艺学报 ›› 2004, Vol. 31 ›› Issue (1): 64-68.

• 研究论文 • 上一篇    下一篇

铁皮石斛类原球茎空气干燥法超低温保存中的脱水蛋白分析

林伟强;边红武;王君晖*;朱睦元   

  1. (浙江大学生命科学学院, 杭州310012)
  • 收稿日期:2003-05-06 修回日期:2003-07-14 出版日期:2004-02-25 发布日期:2004-02-25

Analysis of Dehydrins in Cryopreservation of Protocorm-like-bodies of Dendrobium candidum by the Air-drying Method

Lin Weiqiang;Bian Hongwu ;Wang Junhui*; Zhu Muyuan   

  1. ( College of Life Sciences , Zhejiang University , Hangzhou 310012 , China)
  • Received:2003-05-06 Revised:2003-07-14 Online:2004-02-25 Published:2004-02-25

摘要: 用空气干燥法将铁皮石斛类原球茎脱水至不同含水量, 进行超低温保存和热稳定蛋白的SDS-
PAGE 分析, 并进行脱水蛋白的Western-blot 检测。结果表明: 在本试验条件下, 含水量以g/ g DM表示时,铁皮石斛类原球茎脱水时间(x) 对含水量(y) 的回归方程式为y = - 0.1208 x + 9.9636。含水量下降为0.1~0.5 g/ g DM时, 冻后存活率达到最大值。当含水量达1.0 g/ g DM时, 热稳定蛋白的含量明显增加, 经免疫检测其中分子量分别为28.7 和34.3 kD 的两个热稳定蛋白为脱水蛋白。认为含水量是铁皮石斛超低温保存成功的关键因素, 也是引起脱水蛋白大量积累的重要条件。

关键词: 铁皮石斛, 类原球茎, 脱水蛋白, 含水量, 超低温保存

Abstract: Protocorm2like2bodies of ( PLBs) Dendrobium candidum were desiccated to different water contents by an air-drying method. Samples at different water contents were cryoperserved , and the heat-stable proteins of samples were immunological detected by anti-dehydrin antibody. Water content was based on dry mass , its linear equation was expressed as y = - 0.1208 x + 9.9636. The optimal water content for cryopreservation seemed to be at the range of 0.1 g/ g DM to 0.5 g/ g DM. The amount of heat-stable proteins increased significantly when water content decreased to 1.0 g/ g DM. Results from immunological detection showed that two bands of the heat-stable proteins with respective molecular masses of 28.7 and 34.3 kD were dehydrins , which appeared when water content dropped to 1.0 g/ g DM. Our work revealed that water content might be a key factor for successful cryopreservation as well as for accumulation of dehydrins.

Key words: Dendrobium candidum, Protocorm-like-bodies, Dehydrin, Water content, Cryopreservation

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