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包埋脱水法超低温保存萱藻(Scytosiphon lomentaria)丝状体的研究
庄琰,宫相忠,高伟,张文健
中国海洋大学海洋生命学院 青岛 266003,中国海洋大学海洋生命学院 青岛 266003,中国海洋大学海洋生命学院 青岛 266003,中国海洋大学海洋生命学院 青岛 266003
摘要:
以本实验室保存的萱藻(Scytosiphon lomentaria)丝状体为材料, 用包埋脱水法超低温保存萱藻丝状体, 研究了蔗糖预培养浓度和时间、胶球含水量、化冻温度以及胶球恢复时间对萱藻丝状体存活率和冻存后生长发育能力的影响。结果显示: (1) 萱藻丝状体胶球在0.4mol/L的蔗糖溶液中预培养6h能获得最高的存活率; (2) 萱藻丝状体胶球的最佳含水量较低, 为15%左右, 当含水量高于27%时, 冻存后的存活率为0; (3) 40°C为最适的化冻温度; (4) 化冻后, 将萱藻丝状体胶球在黑暗条件下恢复18h能显著提高存活率; (5) 在最佳条件下, 萱藻丝状体经冻存后的存活率最高可达54.79%, 恢复后的丝状体与冻存前丝状体并无区别, 且经诱导后能够产生正常的孢子囊, 放散的孢子可以发育成叶状体。
关键词:  萱藻(Scytosiphon lomentaria)  丝状体  包埋脱水法  超低温保存
DOI:10.11693/hyhz20131200204
分类号:
基金项目:国家高技术研究发展计划(863 计划), 2012AA10A413 号
CRYOPRESERVATION OF FILAMENTS OF SCYTOSIPHON LOMENTARIA BY ENCAPSULATION-DEHYDRATION
zhuang yan,gong xiangzhong,gao wei and zhang wenjian
ocean university of china,ocean university of china,ocean university of china,ocean university of china
Abstract:
Filaments of Scytosiphon lomentaria cyropreserved were used as the materials to study the impacts of encapsulation-dehydration in terms of sucrose concentration, preculture time, water content of the beads, thawing temperature and the recovery time of the beads, on the survival rate and the growth performance of the filaments. The results are followed. The beads of S. lomentaria had highest survival rate after precultured in 0.4mol/L sucrose for 6 hours. The optimal water content of the beads was relatively low, only about 15%; the survival rate was zero when water content was higher than 27%. Temperature at 40°C was best for thawing the beads. The survival rate of S. lomentaria thawed could be enhanced significantly after recovery in dark for 18h. Under the optimal conditions, the survival rate of the filaments thawed could reach as high as 54.79%, and the recovered filaments were the same to those before cryopreservation. The induced filaments could produce normal sporangia, and the spores released from the sporangia were able to grow into thalli.
Key words:  Scytosiphon lomentaria  filaments  encapsulation-dehydration  cryopreservation
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