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Cryopreservation methods are effective for long-term storage of Labyrinthula cultures

机译:冷冻保存方法对于迷宫培养物的长期保存有效

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Marine heterotrophic protists of the Labyrinthulomycota are of interest for their biotechnological (e.g. thraustochytrid production of lipids) and ecological (e.g. wasting disease and rapid blight by pathogens of the genus Labyrinthula) applications; culture-based laboratory studies are a central technique of this research. However, maintaining such microorganism cultures can be labour- and cost-intensive, with a high risk of culture contamination and die-off over time. Deep-freeze storage, or cryopreservation, can be used to maintain culture back-ups, as well as to preserve the genetic and phenotypic properties of the microorganisms; however, this method has not been tested for the ubiquitous marine protists Labyrinthula spp. In this study, we trialled 12 cryopreservation protocols on 3 Labyrinthula sp. isolates of varying colony morphological traits. After 6 mo at -80?°C storage, the DMSO and glycerol protocols were the most effective cryoprotectants compared to methanol (up to 90% success vs. 50% success, respectively). The addition of 30% horse serum to the cryoprotectant solution increased Labyrinthula sp. growth success by 20-30%. We expect that these protocols will provide extra security for culture-based studies, as well as opportunities for long-term research on key Labyrinthula sp. isolates.
机译:迷路藻菌的海洋异养原生生物因其生物技术(例如脂质的破囊壶菌生产)和生态学(例如浪费疾病和迷路菌属病原体的快速枯萎)而受到关注;基于文化的实验室研究是这项研究的核心技术。然而,维持这种微生物培养可能是劳动和成本密集的,随着时间的流逝,培养物污染和死亡的风险很高。可以使用深冻保存或冷冻保存来维持培养备用,并保留微生物的遗传和表型特性。但是,该方法尚未针对普遍存在的海洋生物Labyrinthula spp进行过测试。在这项研究中,我们在3个迷宫藻上试验了12种冷冻保存方案。不同菌落形态特征的分离株。在-80°C下储存6个月后,与甲醇相比,DMSO和甘油方案是最有效的冷冻保护剂(分别成功率高达90%和50%)。向防冻剂溶液中添加30%的马血清会增加迷宫菌。增长成功20-30%。我们希望这些协议将为基于文化的研究提供额外的安全性,并为关键迷宫菌的长期研究提供机会。隔离株。

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