Cryopreservation of gametophytic thalli of Porphyra yezoensis (Rhodophyceae) by one-step fast cooling
Cryopreservation of gametophytic thalli of Porphyra yezoensis (Rhodophyceae) by one-step fast cooling
复制标题
一步快速冷却法超低温保存条斑紫菜（红藻）配子体菌体
DOI：
10.1007/s10811-012-9887-0
复制
发表时间：
2013
影响因子：
3.3
通讯作者：
Kazuyoshi Kuwano
中科院分区：
文献类型：
--
作者：
Youn Hee Choi;Taek Jeong Nam;Kazuyoshi Kuwano
为建立条斑紫菜配子体一步快速冷冻保存技术，研究了不同冷冻保护剂的毒性和效果。二甲基亚砜（DMSO）在浸泡30秒后，浓度高达30%时无害。然而，超过40%的浓度增加是严重有害的，并且在60 s的浸泡的情况下，在较低浓度下发生有害影响。在所测试的冷冻保护剂中，二甘油是危害最小的，并且在高达50%的浓度下未发现其有害影响，即使在浸泡60 s后也是如此。在各种冷冻保护剂溶液中浸泡30秒后，将装载到吸管中的菌体直接投入液氮中。当向双甘油中加入5%DMSO时，在15- 25%的双甘油浓度下实现了高存活率（超过80%），并且在20%的双甘油浓度下最大存活率为94.6%。解冻后的菌体重新开始生长，与未冻菌体相比没有任何差异。
This study investigated the toxicity and effectiveness of various cryoprotectant solutions to develop a cryopreservation technique by one-step fast cooling for gametophytic thalli ofPorphyra yezoensis. Dimethyl sulfoxide (DMSO) was not harmful at concentrations up to 30 % after 30 s of immersion. However, an increase in concentration beyond 40 % was seriously detrimental, and the detrimental effect occurred at lower concentrations in the case of 60 s of immersion. Diglycerol was the least harmful among cryoprotectants tested, and its harmful effect was not found at concentrations up to 50 %, even after 60 s of immersion. The thalli loaded into a straw were directly plunged into liquid nitrogen after 30 s of immersion in various cryoprotectant solutions. When 5 % DMSO was added to diglycerol, high survival rates (more than 80 %) were achieved at diglycerol concentrations of 15–25 %, and the maximum survival was 94.6 % at diglycerol concentrations of 20 %. The thawed thalli restarted growth without any difference in comparison with unfrozen thalli.