Cryopreservation of gametophytes of Laminaria japonica (Phaeophyta) using encapsulation-dehydration with two-step cooling method

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• 作者：Quansheng Zhang (1) (2)
  Yizhou Cong (3)
  Shancun Qu (3)
  Shiju Luo (3)
  Guanpin Yang (1)
  
• 关键词：Laminaria japonica ; gametophyte ; encapsulation ; dehydration with two ; step cooling ; cryopreservation
• 刊名：Journal of Ocean University of China
• 出版年：2008
• 出版时间：February 2008
• 年：2008
• 卷：7
• 期：1
• 页码：65-71
• 全文大小：755KB
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• 作者单位：Quansheng Zhang (1) (2)
  Yizhou Cong (3)
  Shancun Qu (3)
  Shiju Luo (3)
  Guanpin Yang (1)
  
  1. College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, P. R. China
  2. Ocean School, Yantai University, Yantai, 264005, P. R. China
  3. Shandong Oriental Ocean Sci-tech Co., Ltd., Yantai, 264005, P. R. China
  
• ISSN：1993-5021

文摘

Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10°C and under continuous irradiance of 30 μmol m? s? for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molL? sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10°C and 70% relative humidity for 4 h, pre-frozen at either ?0°C or ?0°C for 30 min, and stored in liquid nitrogen for >24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40°C seawater and re-hydrated in 0.05 molL? citrate sodium prepared using 30-NaCl 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.