Gwo Jin-Chywan, Chiu Ju-Yu, Chou Chin-Cheng, Cheng Hsien-Yu
Department of Aquaculture, Taiwan National Ocean University, Keelung 20224, Taiwan.
Cryobiology. 2005 Jun;50(3):338-43. doi: 10.1016/j.cryobiol.2005.02.001. Epub 2005 Apr 7.
The cryopreservation of algae could prevent genetic drift and minimize labor costs compared to the current method of maintenance and subculturing. Clear, simple protocols for cryopreservation of marine microalga, Nannochloropsis oculata were developed and cryoprotectant choice and concentration optimized. The viability of the microalga was assessed directly after thawing, and algal concentration was measured after 2-30 days of growth. Five cryoprotectants (dimethyl sulphoxide, Me2SO; ethylene glycol, EG; glycerol, Gly; methanol, MeOH; and propylene glycol, PG) at five concentrations (10, 20, 30, 40, and 50%; v/v) were evaluated to determine the toxicity of various cryoprotectants to N. oculata. The toxicity of cryoprotectant (Me2SO, EG, MeOH, and PG) was observed only at higher concentrations of CPAs: > 20% for EG, > 30% for Me2SO and methanol, and > 40% for PG. Direct freezing of algae in liquid nitrogen resulted in a severe loss of viability and a modified cryopreservation protocol proved to be more appropriate for the preservation of N. oculata. Cryopreservation protocols developed and tested in the present study might be applied to cryopreserving other strains, or species, in this genus.
与目前的藻类保存和继代培养方法相比，藻类的冷冻保存可以防止基因漂移并将劳动力成本降至最低。我们制定了清晰、简单的海洋微藻——眼点拟微球藻的冷冻保存方案，并对冷冻保护剂的选择和浓度进行了优化。微藻解冻后直接评估其活力，并在生长2至30天后测量藻类浓度。我们评估了五种浓度（10%、20%、30%、40%和50%；v/v）的五种冷冻保护剂（二甲基亚砜、Me2SO；乙二醇、EG；甘油、Gly；甲醇、MeOH；和丙二醇、PG），以确定各种冷冻保护剂对眼点拟微球藻的毒性。仅在较高浓度的冷冻保护剂下观察到冷冻保护剂（Me2SO、EG、MeOH和PG）的毒性：EG > 20%、Me2SO和甲醇 > 30%、PG > 40%。将藻类直接在液氮中冷冻导致活力严重丧失，而改良的冷冻保存方案被证明更适合眼点拟微球藻的保存。本研究中开发和测试的冷冻保存方案可能适用于该属其他菌株或物种的冷冻保存。
