马来西亚淡水微藻菌株的低温保存方法

A. Afiqah, Siti Saizah, R. Eileen
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引用次数: 0

摘要

微藻在许多生物技术开发中在生产有价值的产品、服务和过程中发挥着重要作用。低温保存技术可用于长期保存许多能在低温保存后存活的微藻菌株。在本研究中,利用小球藻(Chlorella vulgaris),采用三种不同的方案来确定不同浓度的冷冻保护剂对淡水微藻菌株的保存效果。在微藻悬浮液中分别添加0%、10%、20%和30%浓度的二甲亚砜(DMSO)。方案1涉及直接投入液氮,方案2缓慢冷却至-80°C,方案3缓慢冷却至-20°C。以540nm处的吸光度值测定冷冻后的生长情况。方案2中30% DMSO记录的吸光度值最高,为0.387 + 0.015,与方案1和方案3相比,p<0.05显著。普通小球藻低温保存后生长的最佳方案为方案2,以30% DMSO作为冷冻保护剂,缓慢冷却至-80 °C。
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A Cryopreservation Protocol for the Malaysian Fresh Water Microalgal Strain
Microalgae is important in many biotechnological exploitations in producing valuable products, services and processes. The technology of cryopreservation is useful in the long-term storage of many microalgal strains able to survive post cryopreservation. In this study, Chlorella vulgaris was utilised to determine the effects of different concentrations of a cryoprotectant to preserve the fresh water microalgal strain employing three types of protocol. Dimethyl sulfoxide (DMSO) at different concentrations of 0%, 10%, 20% and 30% were added to the microalgal suspension in three types of protocol. Protocol 1 involved direct plunging into liquid nitrogen, Protocol 2 is slow cooling to -80°C and Protocol 3 is slow cooling to -20°C. The absorbance value at 540 nm was used as a measurement to determine the growth post cryopreservation. The absorbance value recorded with 30% DMSO in Protocol 2 was the highest at 0.387 + 0.015 and was significant at p<0.05 compared to Protocol 1 and 3. The best protocol for post cryopreservation growth of Chlorella vulgaris was Protocol 2 using slow cooling to -80 °C treated with 30% DMSO as the cryoprotective agent.
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