Ficetola Gentile Francesco, Miaud Claude, Pompanon François, Taberlet Pierre
Laboratoire d'Ecologie Alpine, CNRS-UMR 5553, Université Joseph Fourier, Grenoble Cedex 09, France.
Biol Lett. 2008 Aug 23;4(4):423-5. doi: 10.1098/rsbl.2008.0118.
The assessment of species distribution is a first critical phase of biodiversity studies and is necessary to many disciplines such as biogeography, conservation biology and ecology. However, several species are difficult to detect, especially during particular time periods or developmental stages, potentially biasing study outcomes. Here we present a novel approach, based on the limited persistence of DNA in the environment, to detect the presence of a species in fresh water. We used specific primers that amplify short mitochondrial DNA sequences to track the presence of a frog (Rana catesbeiana) in controlled environments and natural wetlands. A multi-sampling approach allowed for species detection in all environments where it was present, even at low densities. The reliability of the results was demonstrated by the identification of amplified DNA fragments, using traditional sequencing and parallel pyrosequencing techniques. As the environment can retain the molecular imprint of inhabiting species, our approach allows the reliable detection of secretive organisms in wetlands without direct observation. Combined with massive sequencing and the development of DNA barcodes that enable species identification, this approach opens new perspectives for the assessment of current biodiversity from environmental samples.
物种分布评估是生物多样性研究的首个关键阶段,对生物地理学、保护生物学和生态学等许多学科而言都至关重要。然而,有几种物种难以被检测到,尤其是在特定时间段或发育阶段,这可能会使研究结果产生偏差。在此,我们基于环境中DNA的有限持久性,提出一种新颖的方法来检测淡水中某一物种的存在。我们使用特定引物扩增线粒体短DNA序列,以追踪牛蛙(Rana catesbeiana)在受控环境和天然湿地中的存在情况。采用多次采样方法能够在该物种存在的所有环境中检测到它,即便其密度很低。通过使用传统测序和平行焦磷酸测序技术鉴定扩增的DNA片段,证明了结果的可靠性。由于环境能够保留栖息物种的分子印记,我们的方法无需直接观察就能可靠地检测湿地中的隐匿生物。结合大规模测序以及能够进行物种鉴定的DNA条形码的开发,这种方法为从环境样本评估当前生物多样性开辟了新的前景。