Quince Christopher, Curtis Thomas P, Sloan William T
Department of Civil Engineering, University of Glasgow, Glasgow, UK.
ISME J. 2008 Oct;2(10):997-1006. doi: 10.1038/ismej.2008.69. Epub 2008 Jul 24.
The exploration of the microbial world has been an exciting series of unanticipated discoveries despite being largely uninformed by rational estimates of the magnitude of task confronting us. However, in the long term, more structured surveys can be achieved by estimating the diversity of microbial communities and the effort required to describe them. The rates of recovery of new microbial taxa in very large samples suggest that many more taxa remain to be discovered in soils and the oceans. We apply a robust statistical method to large gene sequence libraries from these environments to estimate both diversity and the sequencing effort required to obtain a given fraction of that diversity. In the upper ocean, we predict some 1400 phylotypes, and a mere fivefold increase in shotgun reads could yield 90% of the metagenome, that is, all genes from all taxa. However, at deep ocean, hydrothermal vents and diversities in soils can be up to two orders of magnitude larger, and hundreds of times the current number of samples will be required just to obtain 90% of the taxonomic diversity based on 3% difference in 16S rDNA. Obtaining 90% of the metagenome will require tens of thousands of times the current sequencing effort. Although the definitive sequencing of hyperdiverse environments is not yet possible, we can, using taxa-abundance distributions, begin to plan and develop the required methods and strategies. This would initiate a new phase in the exploration of the microbial world.
尽管在很大程度上我们对面临任务的规模缺乏合理估计,但对微生物世界的探索仍是一系列令人兴奋的意外发现。然而,从长远来看,通过估计微生物群落的多样性以及描述它们所需的工作量,可以实现更有组织的调查。在非常大的样本中新微生物类群的恢复率表明,土壤和海洋中仍有更多类群有待发现。我们将一种强大的统计方法应用于来自这些环境的大型基因序列文库,以估计多样性以及获得该多样性给定比例所需的测序工作量。在海洋上层,我们预测约有1400个系统发育型,霰弹枪测序读数仅增加五倍就能产生宏基因组的90%,即所有类群的所有基因。然而,在深海、热液喷口和土壤中的多样性可能大两个数量级,仅基于16S rDNA 3%的差异,要获得90%的分类多样性就需要数百倍于当前样本数量的样本。要获得90%的宏基因组将需要数万倍于当前的测序工作量。虽然对超多样环境进行最终测序目前还不可能,但我们可以利用类群丰度分布开始规划和开发所需的方法和策略。这将开启微生物世界探索的新阶段。