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用于从环境DNA中检测难以捉摸的水禽的优化PCR检测方法。

Optimised PCR assays for detecting elusive waterfowl from environmental DNA.

作者信息

Honka Johanna, Kvist Laura, Olli Suvi, Laaksonen Toni, Aspi Jouni

机构信息

Ecology and Genetics Research Unit University of Oulu Oulu Finland.

Department of Biology University of Turku Turku Finland.

出版信息

Ecol Evol. 2024 Apr 1;14(4):e11224. doi: 10.1002/ece3.11224. eCollection 2024 Apr.

Abstract

For many aquatic and semiaquatic mammal, amphibian and fish species, environmental DNA (eDNA) methods are employed to detect species distribution and to monitor their presence, but eDNA is much less employed for avian species. Here, we developed primers for the detection of true geese and swan species using eDNA and optimised a PCR protocol for eDNA. We selected taiga bean goose () as our focal (sub)species and sampled water from lakes, from which the presence of taiga bean goose was visually confirmed. To test, if taiga bean goose DNA could be detected among DNA of other goose species, we similarly sampled eDNA from a zoo pond housing several Anatidae species. We were able to detect taiga bean goose DNA in all but one of the tested lakes, including the zoo pond. The primers developed are not species-specific, but rather specific to the genus , due to the close relatedness of species, which prevented the development of species-specific primers and the use of, for example, quantitative PCR. We also developed eDNA primers for species and species and tested these primers using the same samples. Canada goose () and barnacle goose () DNA were only detected in the zoo pond (in which they were present), as the sampled natural lakes fall outside the range of these species. We detected whooper swan () DNA in three lakes and the zoo pond (in which the species was present). The eDNA method presented here provides a potential means to monitor elusive goose species and to study the co-occurrence of large waterfowl.

摘要

对于许多水生和半水生的哺乳动物、两栖动物及鱼类物种,环境DNA(eDNA)方法被用于检测物种分布并监测它们的存在情况,但eDNA在鸟类物种中的应用要少得多。在此,我们开发了用于通过eDNA检测真鹅和天鹅物种的引物,并优化了一种用于eDNA的PCR方案。我们选择了豆雁()作为我们的重点(亚)物种,并从湖泊中采集水样,在这些湖泊中肉眼确认了豆雁的存在。为了测试是否能在其他鹅类物种的DNA中检测到豆雁的DNA,我们同样从饲养了几种鸭科物种的动物园池塘中采集了eDNA样本。除了一个测试湖泊外,我们在所有测试湖泊中都检测到了豆雁的DNA,包括动物园池塘。由于鹅类物种之间亲缘关系密切,所开发的引物不是物种特异性的,而是属特异性的,这阻碍了物种特异性引物的开发以及例如定量PCR的使用。我们还为物种和物种开发了eDNA引物,并使用相同的样本对这些引物进行了测试。加拿大雁()和黑雁()的DNA仅在动物园池塘(它们存在于其中)中被检测到,因为所采集的天然湖泊不在这些物种的分布范围内。我们在三个湖泊和动物园池塘(该物种存在于其中)中检测到了大天鹅()的DNA。本文介绍的eDNA方法为监测难以捉摸的鹅类物种以及研究大型水禽的共存提供了一种潜在手段。

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