Cryopreservation of Gametophytes of Laminaria japonica (Phaeophyta) Using Encapsulation-Dehydration with Two-Step Cooling Method  

作　　者：ZHANG Quansheng CONG Yizhou QU Shancun LUO Shiju YANG Guanpin 

机构地区：[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

出　　处：《Journal of Ocean University of China》2008年第1期65-71,共7页中国海洋大学学报（英文版）

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 molLl 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‰ 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.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 en- capsulated 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‰ NaCl 7 d later. More cells of male gametophytes sur- vived 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：Laminaria japonica gametophyte encapsulation-dehydration with two-step cooling CRYOPRESERVATION 

分 类 号：Q178.53[生物学—水生生物学]