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六种共生藻属的冷冻保存及其对盐度升高处理的脂肪酸谱的评估。

Cryopreservation of six Symbiodiniaceae genera and assessment of fatty acid profiles in response to increased salinity treatments.

机构信息

Cawthron Institute, Private Bag 2, Nelson, 7042, New Zealand.

School of Biological Sciences, Victoria University of Wellington, PO Box 600, Wellington, 6140, New Zealand.

出版信息

Sci Rep. 2022 Jul 20;12(1):12408. doi: 10.1038/s41598-022-16735-w.

DOI:10.1038/s41598-022-16735-w
PMID:35859115
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9300622/
Abstract

Symbiodiniaceae are a diverse group of dinoflagellates, the majority of which are free-living and/or associated with a variety of protists and other invertebrate hosts. Maintenance of isolated cultures is labour-intensive and expensive, and cryopreservation provides an excellent avenue for their long-term storage. We aimed to cryopreserve 15 cultured isolates from six Symbiodiniaceae genera using dimethyl sulfoxide (DMSO) as the cryoprotectant agent (CPA). Under 15% DMSO, 10 isolates were successfully cryopreserved using either rapid freezing or controlled-rate freezing. Cultures that failed or had low survival, were subjected to (1) a reduction of CPA to 10%, or (2) increased salinity treatment before freezing. At 10% DMSO, three further isolates were successfully cryopreserved. At 15% DMSO there were high cell viabilities in Symbiodinium pilosum treated with 44 parts per thousand (ppt) and 54 ppt culture medium. An isolate of Fugacium sp. successfully cryopreserved after salinity treatments of 54 ppt and 64 ppt. Fatty acid (FA) analyses of S. pilosum after 54 ppt salinity treatment showed increased saturated FA levels, whereas Fugacium sp. had low poly-unsaturated FAs compared to normal salinity (34 ppt). Understanding the effects of salinity and roles of FAs in cryopreservation will help in developing protocols for these ecologically important taxa.

摘要

虫黄藻是一类多样性的甲藻，其中大多数为自由生活或与各种原生生物和其他无脊椎动物宿主相关联。分离培养物的维持需要大量的劳动力和资金，而冷冻保存则为其长期储存提供了一个极好的途径。我们的目的是使用二甲亚砜 (DMSO) 作为冷冻保护剂 (CPA) 来冷冻保存来自六个虫黄藻属的 15 个培养物分离物。在 15% DMSO 下，使用快速冷冻或控速冷冻成功冷冻保存了 10 个分离物。未能成功或存活率低的培养物，经过（1）降低 CPA 至 10%，或（2）冷冻前增加盐度处理。在 10% DMSO 下，又成功冷冻保存了另外三个分离物。在 15% DMSO 下，处理 44 个千分比 (ppt) 和 54 ppt 培养基的 S. pilosum 细胞具有高细胞活力。Fugacium sp. 经过 54 ppt 和 64 ppt 盐度处理后成功冷冻保存。对经过 54 ppt 盐度处理的 S. pilosum 的脂肪酸 (FA) 分析表明，饱和 FA 水平增加，而 Fugacium sp. 的多不饱和 FA 水平较低与正常盐度（34 ppt）相比。了解盐度的影响和 FA 在冷冻保存中的作用将有助于为这些具有生态重要性的分类群开发方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/e8188be4b706/41598_2022_16735_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/ce92c7a733f8/41598_2022_16735_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/397bdf8c79c5/41598_2022_16735_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/3af31d6e9faa/41598_2022_16735_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/e3542b66b4af/41598_2022_16735_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/e8188be4b706/41598_2022_16735_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/ce92c7a733f8/41598_2022_16735_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/397bdf8c79c5/41598_2022_16735_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/3af31d6e9faa/41598_2022_16735_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/e3542b66b4af/41598_2022_16735_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d07/9300622/e8188be4b706/41598_2022_16735_Fig5_HTML.jpg

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