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快速被动收集环境 DNA 的材料比较。

Comparison of materials for rapid passive collection of environmental DNA.

机构信息

Commonwealth Scientific and Industrial Research Organisation, Indian Oceans Marine Research Centre, Oceans and Atmosphere, Crawley, Western Australia, Australia.

Commonwealth Scientific and Industrial Research Organization, Indian Oceans Marine Research Centre, Environomics Future Science Platform, Crawley, Western Australia, Australia.

出版信息

Mol Ecol Resour. 2022 Oct;22(7):2559-2572. doi: 10.1111/1755-0998.13640. Epub 2022 Jun 1.

Abstract

Passive collection is an emerging sampling method for environmental DNA (eDNA) in aquatic systems. Passive eDNA collection is inexpensive and efficient, and requires minimal equipment, making it suited to high-density sampling and remote deployment. Here, we compare the effectiveness of nine membrane materials for passively collecting fish eDNA from a 3-million-litre marine mesocosm. We submerged materials (cellulose, cellulose with 1% and 3% chitosan, cellulose overlayed with electrospun nanofibres and 1% chitosan, cotton fibres, hemp fibres, and sponge with either zeolite or active carbon) for intervals between 5 and 1080 min. We show that for most materials, with as little as 5 min of submersion, mitochondrial fish eDNA measured with qPCR, and fish species richness measured with metabarcoding, was comparable to that collected by conventional filtering. Furthermore, PCR template DNA concentrations and species richness were generally not improved significantly by longer submersion. Species richness detected for all materials ranged between 11 and 37 species, with a median of 27, which was comparable to the range for filtered eDNA (19-32). Using scanning electron microscopy, we visualized biological matter adhering to the surface of materials, rather than entrapped, with images also revealing a diversity in size and structure of putative eDNA particles. eDNA can be collected rapidly from seawater with a passive approach and using a variety of materials. This will suit cost- and time-sensitive biological surveys, and where access to equipment is limited.

摘要

被动采集是一种新兴的水生环境 DNA(eDNA)采样方法。被动 eDNA 采集成本低廉、效率高,所需设备最少,非常适合高密度采样和远程部署。在这里,我们比较了九种膜材料从 300 万升海洋中采集鱼类 eDNA 的效果。我们将材料(纤维素、含 1%和 3%壳聚糖的纤维素、覆盖有静电纺纳米纤维和 1%壳聚糖的纤维素、棉纤维、麻纤维以及沸石或活性炭海绵)浸入水中 5 至 1080 分钟。结果表明,对于大多数材料,只要浸入水中 5 分钟,用 qPCR 测量的线粒体鱼类 eDNA 和用 metabarcoding 测量的鱼类物种丰富度,与传统过滤法收集的 eDNA 相当。此外,更长时间的浸入并不会显著提高 PCR 模板 DNA 浓度和物种丰富度。所有材料检测到的物种丰富度在 11 到 37 种之间,中位数为 27,与过滤的 eDNA(19-32)相当。通过扫描电子显微镜,我们观察到生物物质粘附在材料表面,而不是被困在材料中,图像还揭示了 eDNA 颗粒的大小和结构多样性。通过被动方法和使用多种材料,可以从海水中快速采集 eDNA。这将适合于成本敏感和时间敏感的生物调查,以及设备有限的情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6868/9544503/ea1af06728e1/MEN-22-2559-g005.jpg

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