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Cryopreservation of Gametophytes of Laminaria japonica (Phaeophyta) Using Encapsulation-Dehydration with Two-Step Cooling Method
作者姓名:ZHANG  Quansheng  CONG  Yizhou  QU  Shancun  LUO  Shiju  YANG  Guanpin
作者单位:[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
基金项目:国家重点基础研究发展计划(973计划),山东省科技发展基金
摘    要:Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m-2 s-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molL-1 sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ 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℃seawater and re-hydrated in 0.05 molL-1 citrate sodium prepared using 30‰ NaC1 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.

关 键 词:昆布属植物  配偶体  冷却方法  海洋植物
收稿时间:2006-11-09
修稿时间:2007-08-03

Cryopreservation of gametophytes of Laminaria japonica (Phaeophyta) using encapsulation-dehydration with two-step cooling method
ZHANG Quansheng CONG Yizhou QU Shancun LUO Shiju YANG Guanpin.Cryopreservation of gametophytes of Laminaria japonica (Phaeophyta) using encapsulation-dehydration with two-step cooling method[J].Journal of Ocean University of China,2008,7(1):65-71.
Authors:Quansheng Zhang  Yizhou Cong  Shancun Qu  Shiju Luo  Guanpin Yang
Institution:1. College of Marine Life Sciences, Ocean University of China, Qingdao 266003, P. R. China;Ocean School, Yantai University, Yantai 264005, P. R. China
2. Shandong Oriental Ocean Sci-tech Co., Ltd., Yantai 264005, P. R. China
3. College of Marine Life Sciences, Ocean University of China, Qingdao 266003, P. R. China
Abstract: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−2 s−1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molL−1 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 −40°C or −60°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−1 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.
Keywords:Laminaria japonica  gametophyte  encapsulation-dehydration with two-step cooling  cryopreservation
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