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金属有机框架中环境DNA的捕获与保护

Capture and Protection of Environmental DNA in a Metal-Organic Framework.

作者信息

FitzGerald Laura I, Hahn Erin E, Wallace Mark, Stephenson Sarah A, Berry Oliver F, Doherty Cara M

机构信息

Manufacturing CSIRO Clayton South 3169 VIC Australia.

National Research Collections Australia CSIRO Canberra 2601 ACT Australia.

出版信息

Small Sci. 2024 Oct 1;4(12):2400432. doi: 10.1002/smsc.202400432. eCollection 2024 Dec.

Abstract

Environmental DNA (eDNA) is released by organisms into their surroundings, enabling non-invasive species detection and biodiversity assessments without the need for direct observation. However, collection poses challenges due to the generally low abundance of eDNA and the presence of degradation agents, including enzymes, UV radiation, and microorganisms, rendering samples unstable. Active filtration, which is frequently used to capture eDNA in water, can be time-consuming and cumbersome in field conditions. Herein, a filter-free one-pot procedure for capturing eDNA with the metal-organic framework (MOF), zeolitic imidazolate framework 8 (ZIF-8), is examined. The method is evaluated on 15 mL water samples from diverse sources (aquarium, river, and sea). ZIF-8 forms in all with high capture efficiency (>98%) using spiked salmon DNA to represent eDNA. The DNA is resistant to degradation by endonucleases and UV light. In addition, it remains stable over time as a species-specific salmon quantitative polymerase chain reaction detected genomic DNA in all samples captured with the MOF to a maximum of 28 days at 37 °C while the untreated control samples were below the assay detection limit by day 6. These results highlight the efficacy of ZIF-8 capture in overcoming challenges associated with the preservation of eDNA obtained from aquatic environments.

摘要

环境DNA(eDNA)由生物体释放到周围环境中,无需直接观察就能实现非侵入性物种检测和生物多样性评估。然而,由于eDNA的丰度通常较低,且存在包括酶、紫外线辐射和微生物在内的降解因子,导致样本不稳定,样本采集面临挑战。在野外条件下,常用于捕获水中eDNA的主动过滤可能既耗时又麻烦。在此,研究了一种使用金属有机框架(MOF)——沸石咪唑酯骨架8(ZIF-8)捕获eDNA的无过滤一锅法。该方法在来自不同来源(水族馆、河流和海洋)的15毫升水样上进行了评估。使用添加的鲑鱼DNA代表eDNA,ZIF-8在所有情况下均以高捕获效率(>98%)形成。该DNA对内切核酸酶和紫外线具有抗性。此外,随着时间的推移,它保持稳定,因为一种针对特定物种的鲑鱼定量聚合酶链反应在37°C下检测到用MOF捕获的所有样本中的基因组DNA最长可达28天,而未处理的对照样本在第6天就低于检测限。这些结果突出了ZIF-8捕获在克服与从水生环境中获取的eDNA保存相关挑战方面的有效性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1865/11935135/8793b72c82e8/SMSC-4-2400432-g006.jpg

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