Molecular detection of vertebrates in stream water: a demonstration using Rocky Mountain tailed frogs and Idaho giant salamanders.
Molecular detection of vertebrates in stream water: a demonstration using Rocky Mountain tailed frogs and Idaho giant salamanders.
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DOI：
10.1371/journal.pone.0022746
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Waits LP
中科院分区：
文献类型：
--
作者：
Goldberg CS;Pilliod DS;Arkle RS;Waits LP
河流生态系统中隐藏着许多隐秘的濒危物种，对这些系统中脊椎动物的研究面临着相对低的检测率和高成本的挑战。环境DNA（eDNA）最近被证实是一个敏感和有效的工具，记录水生脊椎动物在湿地和大型河流和运河系统。然而，目前还不清楚这种工具是否可以用于检测快速移动的溪流中的低密度脊椎动物，其中脱落的细胞可能会迅速远离其来源。为了评估eDNA技术在溪流系统中的潜在效用，我们设计了靶向引物来扩增美国西北部地区两种分泌性溪流两栖动物物种（落基山尾蛙，Ascaphus montanus和爱达荷州大鲵，Dicamptodon aterrimus）的短的、物种特异性的DNA片段。我们测试了三种DNA提取和五种PCR方案，以确定我们是否可以检测到这些物种的eDNA在过滤后的水样品从五个流与这些物种的不同密度在美国中部爱达荷州。我们成功地扩增和测序的目标DNA区域的两个物种从流水过滤器样品。我们在所有样本中检测到爱达荷州的巨型蝾螈，在五条溪流中的四条中检测到落基山尾蛙，并发现一些迹象表明，这些物种在早春比在初秋更难用eDNA检测到。虽然这种方法在分类群中的灵敏度仍有待确定，但eDNA的使用可能会彻底改变对稀有和入侵物种的调查。通过这项研究，eDNA技术用于检测水生脊椎动物的实用性已在大多数淡水系统中得到证明，为水生研究方法的创新转型奠定了基础。
Stream ecosystems harbor many secretive and imperiled species, and studies of vertebrates in these systems face the challenges of relatively low detection rates and high costs. Environmental DNA (eDNA) has recently been confirmed as a sensitive and efficient tool for documenting aquatic vertebrates in wetlands and in a large river and canal system. However, it was unclear whether this tool could be used to detect low-density vertebrates in fast-moving streams where shed cells may travel rapidly away from their source. To evaluate the potential utility of eDNA techniques in stream systems, we designed targeted primers to amplify a short, species-specific DNA fragment for two secretive stream amphibian species in the northwestern region of the United States (Rocky Mountain tailed frogs, Ascaphus montanus, and Idaho giant salamanders, Dicamptodon aterrimus). We tested three DNA extraction and five PCR protocols to determine whether we could detect eDNA of these species in filtered water samples from five streams with varying densities of these species in central Idaho, USA. We successfully amplified and sequenced the targeted DNA regions for both species from stream water filter samples. We detected Idaho giant salamanders in all samples and Rocky Mountain tailed frogs in four of five streams and found some indication that these species are more difficult to detect using eDNA in early spring than in early fall. While the sensitivity of this method across taxa remains to be determined, the use of eDNA could revolutionize surveys for rare and invasive stream species. With this study, the utility of eDNA techniques for detecting aquatic vertebrates has been demonstrated across the majority of freshwater systems, setting the stage for an innovative transformation in approaches for aquatic research.
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DOI：
10.1111/j.1471-8286.2007.01678.x
发表时间：
2007-05-01
期刊：
Molecular ecology notes
影响因子：
--
作者：
Ratnasingham S;Hebert PD
通讯作者：
Hebert PD
影响因子：
2.6
作者：
Ryu, C;Lee, K;Oh, HB
通讯作者：
Oh, HB
影响因子：
4.9
作者：
Carstens, BC;Degenhardt, JD;Sullivan, J
通讯作者：
Sullivan, J
影响因子：
46.9
作者：
Rothberg, Jonathan M.;Leamon, John H.
通讯作者：
Leamon, John H.
影响因子：
--
作者：
Strayer, David L.;Dudgeon, David
通讯作者：
Dudgeon, David