Department of Arctic and Marine Biology, University of Tromsø, Tromsø, Norway.
Mol Ecol. 2012 Aug;21(15):3647-55. doi: 10.1111/j.1365-294X.2012.05545.x. Epub 2012 Apr 16.
Ecosystems across the globe are threatened by climate change and human activities. New rapid survey approaches for monitoring biodiversity would greatly advance assessment and understanding of these threats. Taking advantage of next-generation DNA sequencing, we tested an approach we call metabarcoding: high-throughput and simultaneous taxa identification based on a very short (usually <100 base pairs) but informative DNA fragment. Short DNA fragments allow the use of degraded DNA from environmental samples. All analyses included amplification using plant-specific versatile primers, sequencing and estimation of taxonomic diversity. We tested in three steps whether degraded DNA from dead material in soil has the potential of efficiently assessing biodiversity in different biomes. First, soil DNA from eight boreal plant communities located in two different vegetation types (meadow and heath) was amplified. Plant diversity detected from boreal soil was highly consistent with plant taxonomic and growth form diversity estimated from conventional above-ground surveys. Second, we assessed DNA persistence using samples from formerly cultivated soils in temperate environments. We found that the number of crop DNA sequences retrieved strongly varied with years since last cultivation, and crop sequences were absent from nearby, uncultivated plots. Third, we assessed the universal applicability of DNA metabarcoding using soil samples from tropical environments: a large proportion of species and families from the study site were efficiently recovered. The results open unprecedented opportunities for large-scale DNA-based biodiversity studies across a range of taxonomic groups using standardized metabarcoding approaches.
全球各地的生态系统都受到气候变化和人类活动的威胁。新的快速生物多样性监测方法将极大地促进对这些威胁的评估和理解。我们利用新一代 DNA 测序技术,测试了一种称为 metabarcoding 的方法:基于非常短(通常<100 个碱基对)但信息量丰富的 DNA 片段进行高通量和同时的分类群识别。短 DNA 片段允许使用环境样本中降解的 DNA。所有分析均包括使用植物特异性通用引物进行扩增、测序和分类多样性估计。我们分三个步骤测试了来自土壤中死亡物质的降解 DNA 是否有潜力有效地评估不同生物群落中的生物多样性。首先,从位于两种不同植被类型(草地和石南地)的八个北方植物群落中扩增了土壤 DNA。从北方土壤中检测到的植物多样性与从常规地上调查中估计的植物分类和生长形式多样性高度一致。其次,我们使用来自温带环境中曾经耕种过的土壤的样本评估 DNA 的持久性。我们发现,回收的作物 DNA 序列数量与最后一次耕种以来的年份强烈相关,而附近未耕种的地块则没有作物序列。第三,我们使用来自热带环境的土壤样本评估了 DNA metabarcoding 的普遍适用性:研究地点的大量物种和科得到了有效回收。这些结果为使用标准化 metabarcoding 方法对一系列分类群进行大规模基于 DNA 的生物多样性研究开辟了前所未有的机会。