响应面法优化普通小球藻低温短期保存条件OA

This study aimed to optimize the short-term cryopreservation protocol for Chlorella vulgaris and improve the post-resuscitation survival rate of cells,so as to provide high-quality biological feed for economically important aquatic animals.With C.vulgaris used as the test strain and glycerol adopted as the cryoprotectant,the preliminary optimal ranges of three key parameters including cryopreservation temperature,algal cell concentration and preservation duration were determined through single-factor experiments,and the protocol was further optimized by response surface methodology.The results showed that the optimal single-factor conditions for cryopreservation of cells are-80℃,5.0×105 cells/mL,and 12 hours,respectively.The three parameters ranked in descending order of their influence on the survival rate of algal cells as follows:preservation duration>storage temperature>algal cell concentration.Through response surface analysis,the theoretically optimal preservation conditions were identified as a preservation duration of 70.179 h,a storage temperature of-78.91℃,and an algal cell concentration of 3.77×105 cells/mL.Considering the operability in practical experiments,the optimized conditions were adjusted to a preservation duration of 70 h,a storage temperature of-80℃and an algal cell concentration of 3.77×10 ⁵ cells/mL.Under this adjusted protocol,the survival rate of C.vulgaris cells reached 70.11%after 24 h of resuscitation.These findings provided a reliable reference for the establishment of cryopreservation protocols for Chlorella vulgaris and other freshwater microalgal cells.