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用包埋脱水法冰冻保存小新月菱形藻
王起华1, 李 莹2, 刘艳萍1, 李大鹏3
1.辽宁师范大学生命科学学院;2.辽阳职业技术学院师范分院;3.中国科学院海洋研究所
摘要:
采用包埋脱水法研究了小新月菱形藻(Nitzschia closterium minutissima)的超低温保存,探讨了影响存活率的内、外因素:藻细胞年龄、脱水速率、胶球含水量和化冻后的恢复方法等。结果表明,将处于静止初 期的藻细胞包埋在含有0.5 mol/L蔗糖的3%褐藻酸钙胶球中,以含水量每小时减少0.9%的平均速率脱水至胶球含水量为40%,化冻后在冻存管中室温下暗恢复12 h的条件下冰冻存活率较高,可达到18.3%左右。与常规的两步法冰冻保存相比,包埋脱水法可大大简化操作程序,是保存小新月菱形藻的一种有潜力的方法。
关键词:  小新月菱形藻(Nitzschia closterium minutissima)  包埋脱水法  冰冻(超低温)保存
DOI:
分类号:
基金项目:国家自然科学基金资助项目(30170099,30470184);辽宁省教育厅科研基金资助项目(20041015)
Cryopreservation of Nitzschia closterium minutissima by encapsulation-dehydration
Abstract:
Nitzschia closterium minutissima was preserved in liquid nitrogen by encapsulation-dehydration. The main factors influencing the algal viabilitiy after cryopreservation, such as the age of algal cells, the dehydration rate, the water content of beads and the recovery method after thawing, were studied. The results showed that the algae viability after cryopreservation reached about 18.3%——a relatively high viable rate when the algal cells in a stationary phase were encapsulated in the beads of 3% Ca-alginate with 0.5 mol/L of sucrose and the beads were dehydrated at an average rate of water content reduction 0.9%/h to 40% of the water content. Compared with the conventional two-step freezing method, the encapsulation-dehydration method simplified the operation procedure of cryopreservation greatly and so did a potential method for preservation of N. closterium minutissima.
Key words:  Nitzschia closterium minutissima  encapsulation-dehydration  cryopreservation
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