O'Donnell James L, Kelly Ryan P, Shelton Andrew Olaf, Samhouri Jameal F, Lowell Natalie C, Williams Gregory D
School of Marine and Environmental Affairs, University of Washington , Seattle , WA , United States of America.
Earth Resource Technology, Inc., Under Contract to the Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration , Seattle , WA , United States of America.
PeerJ. 2017 Feb 28;5:e3044. doi: 10.7717/peerj.3044. eCollection 2017.
In the face of increasing threats to biodiversity, the advancement of methods for surveying biological communities is a major priority for ecologists. Recent advances in molecular biological technologies have made it possible to detect and sequence DNA from environmental samples (environmental DNA or eDNA); however, eDNA techniques have not yet seen widespread adoption as a routine method for biological surveillance primarily due to gaps in our understanding of the dynamics of eDNA in space and time. In order to identify the effective spatial scale of this approach in a dynamic marine environment, we collected marine surface water samples from transects ranging from the intertidal zone to four kilometers from shore. Using PCR primers that target a diverse assemblage of metazoans, we amplified a region of mitochondrial 16S rDNA from the samples and sequenced the products on an Illumina platform in order to detect communities and quantify their spatial patterns using a variety of statistical tools. We find evidence for multiple, discrete eDNA communities in this habitat, and show that these communities decrease in similarity as they become further apart. Offshore communities tend to be richer but less even than those inshore, though diversity was not spatially autocorrelated. Taxon-specific relative abundance coincided with our expectations of spatial distribution in taxa lacking a microscopic, pelagic life-history stage, though most of the taxa detected do not meet these criteria. Finally, we use carefully replicated laboratory procedures to show that laboratory treatments were remarkably similar in most cases, while allowing us to detect a faulty replicate, emphasizing the importance of replication to metabarcoding studies. While there is much work to be done before eDNA techniques can be confidently deployed as a standard method for ecological monitoring, this study serves as a first analysis of diversity at the fine spatial scales relevant to marine ecologists and confirms the promise of eDNA in dynamic environments.
面对生物多样性受到的威胁日益增加,开发生物群落调查方法是生态学家的首要任务。分子生物技术的最新进展使从环境样本(环境DNA或eDNA)中检测DNA并进行测序成为可能;然而,eDNA技术尚未作为生物监测的常规方法得到广泛应用,主要是因为我们对eDNA在空间和时间上的动态变化缺乏了解。为了确定这种方法在动态海洋环境中的有效空间尺度,我们从潮间带至离岸四公里的样带采集了海洋表层水样。使用针对多种后生动物的PCR引物,我们从样本中扩增出线粒体16S rDNA区域,并在Illumina平台上对产物进行测序,以便使用各种统计工具检测群落并量化其空间格局。我们发现该栖息地存在多个离散的eDNA群落,并表明这些群落之间的距离越远,相似性越低。近海群落往往比近海群落更丰富,但均匀度更低,尽管多样性在空间上不存在自相关性。特定分类单元的相对丰度与我们对缺乏微观浮游生活史阶段的分类单元空间分布的预期一致,尽管检测到的大多数分类单元不符合这些标准。最后,我们使用精心重复的实验室程序表明,在大多数情况下,实验室处理非常相似,同时使我们能够检测到一个错误的重复样本,强调了重复对元条形码研究的重要性。虽然在eDNA技术能够作为生态监测的标准方法得到可靠应用之前还有很多工作要做,但这项研究首次分析了与海洋生态学家相关的精细空间尺度上的多样性,并证实了eDNA在动态环境中的前景。