Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host
Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host
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DOI：
10.1371/journal.pone.0211755
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发表时间：
2019-01
期刊：
影响因子：
3.7
通讯作者：
Samantha R. Coy;A. Alsante;J. V. Van Etten;S. Wilhelm
中科院分区：
文献类型：
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作者：
Samantha R. Coy;A. Alsante;J. V. Van Etten;S. Wilhelm
实验室文化管理中的最佳实践通常包括对微生物的冷冻保存，但通过保留病毒隔离的病毒颗粒可以挑战研究人员。开发了一种冷冻保存的方法，即绿色 - 艾尔加感染病毒，囊虫病毒1（PBCV-1）探索了该病毒颗粒的感染的冷冻保存，从而在没有冷冻保护剂的情况下保持最高的感染（〜2.5％）。发现当感染宿主在感染的中期阶段被冷冻保存时，病毒生存率最高（69.5±16.5％）（即与病毒一致组装）。感染后240分钟，颗粒和病毒保存。感染宿主的冷冻保存可能是在环境中不增长的情况下（例如古代冻土）中病毒持久性的可靠机制。
Best practices in laboratory culture management often include cryopreservation of microbiota, but this can be challenging with some virus particles. By preserving viral isolates researchers can mitigate genetic drift and laboratory-induced selection, thereby maintaining genetically consistent strains between experiments. To this end, we developed a method to cryopreserve the model, green-alga infecting virus, Paramecium bursaria Chlorella virus 1 (PBCV-1). We explored cryotolerance of the infectivity of this virus particle, whereby freezing without cryoprotectants was found to maintain the highest infectivity (~2.5%). We then assessed the cryopreservation potential of PBCV-1 during an active infection cycle in its Chlorella variabilis NC64A host, and found that virus survivorship was highest (69.5 ± 16.5 %) when the infected host is cryopreserved during mid-late stages of infection (i.e., coinciding with virion assembly). The most optimal condition for cryopreservation was observed at 240 minutes post-infection. Overall, utilizing the cell as a vehicle for viral cryopreservation resulted in 24.9 – 30.1 fold increases in PBCV-1 survival based on 95% confidence intervals of frozen virus particles and virus cryopreserved at 240 minutes post-infection. Given that cryoprotectants are often naturally produced by psychrophilic organisms, we suspect that cryopreservation of infected hosts may be a reliable mechanism for virus persistence in non-growth permitting circumstances in the environment, such as ancient permafrosts.