深渊中的真核生物多样性:来自焦磷酸测序的新认识。
Eukaryotic richness in the abyss: insights from pyrotag sequencing.
机构信息
Department of Genetics and Evolution, University of Geneva, Geneva, Switzerland.
出版信息
PLoS One. 2011 Apr 4;6(4):e18169. doi: 10.1371/journal.pone.0018169.
BACKGROUND
The deep sea floor is considered one of the most diverse ecosystems on Earth. Recent environmental DNA surveys based on clone libraries of rRNA genes confirm this observation and reveal a high diversity of eukaryotes present in deep-sea sediment samples. However, environmental clone-library surveys yield only a modest number of sequences with which to evaluate the diversity of abyssal eukaryotes.
METHODOLOGY/PRINCIPAL FINDINGS: Here, we examined the richness of eukaryotic DNA in deep Arctic and Southern Ocean samples using massively parallel sequencing of the 18S ribosomal RNA (rRNA) V9 hypervariable region. In very small volumes of sediments, ranging from 0.35 to 0.7 g, we recovered up to 7,499 unique sequences per sample. By clustering sequences having up to 3 differences, we observed from 942 to 1756 Operational Taxonomic Units (OTUs) per sample. Taxonomic analyses of these OTUs showed that DNA of all major groups of eukaryotes is represented at the deep-sea floor. The dinoflagellates, cercozoans, ciliates, and euglenozoans predominate, contributing to 17%, 16%, 10%, and 8% of all assigned OTUs, respectively. Interestingly, many sequences represent photosynthetic taxa or are similar to those reported from the environmental surveys of surface waters. Moreover, each sample contained from 31 to 71 different metazoan OTUs despite the small sample volume collected. This indicates that a significant faction of the eukaryotic DNA sequences likely do not belong to living organisms, but represent either free, extracellular DNA or remains and resting stages of planktonic species.
CONCLUSIONS/SIGNIFICANCE: In view of our study, the deep-sea floor appears as a global DNA repository, which preserves genetic information about organisms living in the sediment, as well as in the water column above it. This information can be used for future monitoring of past and present environmental changes.
背景
深海海底被认为是地球上最多样化的生态系统之一。最近基于 rRNA 基因克隆文库的环境 DNA 调查证实了这一观察结果,并揭示了深海沉积物样本中存在的大量真核生物多样性。然而,环境克隆文库调查只能获得少量序列,用于评估深海真核生物的多样性。
方法/主要发现:在这里,我们使用 18S 核糖体 RNA(rRNA)V9 高变区的大规模平行测序来检查北极和南大洋深部样本中真核生物 DNA 的丰富度。在非常小体积的沉积物中,每个样本的体积从 0.35 克到 0.7 克不等,我们从每个样本中回收了多达 7499 个独特的序列。通过聚类具有多达 3 个差异的序列,我们观察到每个样本中有 942 到 1756 个操作分类单元(OTU)。对这些 OTU 的分类分析表明,所有主要真核生物群体的 DNA 都存在于深海海底。甲藻、原生动物、纤毛虫和眼虫动物占主导地位,分别占所有分配 OTU 的 17%、16%、10%和 8%。有趣的是,许多序列代表光合作用类群,或与从地表水环境调查中报告的序列相似。此外,尽管采集的样本体积很小,但每个样本中仍包含 31 到 71 个不同的后生动物 OTU。这表明,相当一部分真核生物 DNA 序列可能不属于活生物体,而是代表游离的、细胞外的 DNA 或浮游物种的残余物和休眠阶段。
结论/意义:鉴于我们的研究,深海海底似乎是一个全球 DNA 储存库,它保存了生活在沉积物中和其上方水柱中的生物的遗传信息。这些信息可用于未来监测过去和现在的环境变化。