An eDNA approach to detect eastern hellbenders (Cryptobranchus a. alleganiensis) using samples of water
An eDNA approach to detect eastern hellbenders (Cryptobranchus a. alleganiensis) using samples of water
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DOI：
10.1071/wr12114
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发表时间：
2012-01-01
影响因子：
1.9
通讯作者：
Williams, Rod N.
中科院分区：
文献类型：
--
作者：
Olson, Zachary H.;Briggler, Jeffrey T.;Williams, Rod N.
上下文环境DNA或eDNA方法是非侵入性遗传采样的新应用，其中通过水或土壤采样检测来自生物体的DNA，通常用于确定生物体的存在或不存在的目的。eDNA方法有可能彻底改变稀有或濒危类群的研究。我们评估了有效的eDNA采样检测种群的两栖动物保护的关注，东部hellbender（隐鳃亚目。alleganiensis），间接来源于其水生环境。我们开发了物种特异性引物，验证了它们的特异性和灵敏度，并在计算机和实验室试验中评估了我们的方法的实用性。在现场，我们从三个地点与已知的密度hellbenders，并从一个网站，hellbenders不发生的水样。我们过滤水样，从过滤器中提取DNA，并通过聚合酶链反应（PCR）和凝胶电泳分析提取产物中的Helbender DNA。关键结果。我们的方法检测到密度接近最低的报告自然密度的hellbenders。低密度（0.16只/100 m2）的样地有2个阳性扩增，中密度（0.38只/100 m2）的样地有8个阳性扩增，高密度（0.88只/100 m2）的样地有10个阳性扩增。当考虑河流流量时，密度和检测之间的明显关系变得模糊。所有阴性对照均无扩增。eDNA的方法可能是一个具有成本效益的手段，通过它来建立大规模的占有模式的hellbenders. Implications。eDNA可以被认为是一种有价值的工具，用于检测许多难以研究的物种。
Context. Environmental DNA, or eDNA, methods are a novel application of non-invasive genetic sampling in which DNA from organisms is detected via sampling of water or soil, typically for the purposes of determining the presence or absence of an organism. eDNA methods have the potential to revolutionise the study of rare or endangered taxa.Aims. We evaluated the efficacy of eDNA sampling to detect populations of an amphibian of conservation concern, the eastern hellbender (Cryptobranchus a. alleganiensis), indirectly from their aquatic environments.Methods. We developed species-specific primers, validated their specificity and sensitivity, and assessed the utility of our methods in silico and in laboratory trials. In the field, we collected water samples from three sites with known densities of hellbenders, and from one site where hellbenders do not occur. We filtered water samples, extracted DNA from filters, and assayed the extraction products for hellbender DNA by using polymerase chain reaction (PCR) and gel electrophoresis.Key results. Our methods detected hellbenders at densities approaching the lowest of reported natural densities. The low-density site (0.16 hellbenders per 100m(2)) yielded two positive amplifications, the medium-density site (0.38 hellbenders per 100m(2)) yielded eight positive amplifications, and the high-density site (0.88 hellbenders per 100m(2)) yielded 10 positive amplifications. The apparent relationship between density and detection was obfuscated when river discharge was considered. There was no amplification in any negative control.Conclusion. eDNA methods may represent a cost-effective means by which to establish broad-scale patterns of occupancy for hellbenders.Implications. eDNA can be considered a valuable tool for detecting many species that are otherwise difficult to study.