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为建立普通小球藻(Chlorella vulgaris)短期低温保存方法，优化低温保存条件，为水产经济动物提供优质生物饵料，试验以小球藻为受试藻株，甘油作为冷冻保护剂，藻细胞存活率为优化目标，作单因素试验筛选低温保存温度、藻细胞密度和时间共3个条件的最适范围，采用响应面法进一步优化短期低温保存条件并作验证试验。结果表明：通过单因素试验筛选出相对适宜小球藻短期低温保存温度、藻细胞密度和时间分别为-20～-196 ℃、2.5×105～5×105 cell/mL和12～144 h。响应面结果分析表明，对藻细胞存活率影响强弱依次为保存时间、保存温度和藻细胞密度；通过响应面法分析得到上述三个条件的最佳值为70.179 h、-78.91 ℃以及3.77×105 cell/mL；优化后的保存条件为藻细胞密度3.77×105 cell/mL，保存时间70 h，保存温度-80 ℃，小球藻复苏24 h细胞存活率70.11%。优化的条件为小球藻及其他淡水微藻细胞的低温保存提供了技术参考。
Abstract：To develop the short-term cryopreservation protocol for Chlorella vulgaris， optimize the associated conditions to ensure cell viability and provide high-quality biological feed for aquatic economic animals. The study used Chlorella vulgaris as the test strain， glycerol as the cryoprotectant， and the major performance indicator was the algal cell survival rate. After an initial one-factor test to establish the optimal range for short-term cryopreservation temperature， cell density， and duration， a response surface methodology was employed to refine these parameters. A one-factor experiment revealed that the temperature range of -20 ℃ to -196 ℃， the algal cell density of 2.5×105 to 5×105 cells/mL， and the length of 12 to 144 h were the comparatively ideal conditions for the low-temperature preservation of Chlorella vulgaris. Response surface analysis indicated that， in order of significance， preservation period， preservation temperature， and algal cell density all affect the survival rate of algal cells. Cell activity was significantly influenced by preservation duration， temperature， and cell density. The optimal conditions were determined using response surface methods to be 70 h， -78.91℃， with a cell density of 3.77×105 cells/mL. A cell density of 3.77×105 cells/mL， a preservation period of 70 h， and a temperature of -80 ℃ were further determined to be the optimal preservation parameters. After 24 hours， the post-thaw survival rate of algal cells was 70.11%. A technical framework for the cryopreservation of Chlorella and other freshwater microalgae is provided by these ideal conditions， which guarantee excellent cell viability after thawing.
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基本信息:
中图分类号:S963.213
引用信息:
[1]高金伟,张芮,姜青青,等.响应面法优化普通小球藻低温短期保存条件[J].经济动物学报().
2025-05-18
2025-05-18
2025-05-18