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本文将鱼类抗冻蛋白应用于植物细胞的超低温保存。结果表明,在水稻悬浮细胞的两步法保存中,浓度为0.01mg/ml的抗冻蛋白具有特别的负作用,相对高浓度的抗冻蛋白则能减小细胞存活率的波动性。在玻璃化法保存中,浓度为0.2mg/ml的抗冻蛋白能改善保存效果,更高浓度的抗冻蛋白(>5mg/ml)反而会降低保存效果。环境冰晶量、抗冻蛋白浓度、低温保护剂浓度和细胞膜组成等是影响抗冻蛋白使用效果的几大因素。作者在机理分析中认为,一方面,抗冻蛋白能和冰晶作用,抑制重冰晶,防止去玻璃化;另一方面,抗冻蛋白也能和细胞膜作用,诱发膜表面冰晶形成。
In this paper, fish antifreeze protein is applied to the cryopreservation of plant cells. The results showed that antifreeze protein with a concentration of 0.01 mg / ml had a special negative effect in the two-step preservation of rice suspension cells, whereas the relatively high concentration of antifreeze protein could reduce the fluctuation of cell survival rate. Antifreeze protein at a concentration of 0.2 mg / ml can improve the preservative effect in the vitrification method. Higher concentrations of antifreeze protein (> 5 mg / ml) will reduce the preservative effect. Environmental ice volume, antifreeze protein concentration, low temperature protective agent concentration and cell membrane composition are several factors that affect the antifreeze protein use. In the mechanism analysis, the author believes that on the one hand, antifreeze protein can act with ice crystals to inhibit heavy ice crystals and prevent de vitrification; on the other hand, antifreeze proteins can also act on the cell membrane to induce ice crystal formation on the membrane surface.