Opinion: Re-evaluating prokaryotic species

There is no widely accepted concept of species for prokaryotes, and assignment of isolates to species is based on measures of phenotypic or genome similarity. The current methods for defining prokaryotic species are inadequate and incapable of keeping pace with the levels of diversity that are being uncovered in nature. Prokaryotic taxonomy is being influenced by advances in microbial population genetics, ecology and genomics, and by the ease with which sequence data can be obtained. Here, we review the classical approaches to prokaryotic species definition and discuss the current and future impact of multilocus nucleotide-sequence-based approaches to prokaryotic systematics. We also consider the potential, and difficulties, of assigning species status to biologically or ecologically meaningful sequence clusters.     

A Proposed Genus Boundary for the Prokaryotes Based on Genomic Insights

Genomic information has already been applied to prokaryotic species definition and classification. However, the contribution of the genome sequence to prokaryotic genus delimitation has been less studied. To gain insights into genus definition for the prokaryotes, we attempted to reveal the genus-level genomic differences in the current prokaryotic classification system and to delineate the boundary of a genus on the basis of genomic information. The average nucleotide sequence identity between two genomes can be used for prokaryotic species delineation, but it is not suitable for genus demarcation. We used the percentage of conserved proteins (POCP) between two strains to estimate their evolutionary and phenotypic distance. A comprehensive genomic survey indicated that the POCP can serve as a robust genomic index for establishing the genus boundary for prokaryotic groups. Basically, two species belonging to the same genus would share at least half of their proteins. In a specific lineage, the genus and family/order ranks showed slight or no overlap in terms of POCP values. A prokaryotic genus can be defined as a group of species with all pairwise POCP values higher than 50%. Integration of whole-genome data into the current taxonomy system can provide comprehensive information for prokaryotic genus definition and delimitation.     

Environmental genomics, the big picture?

The enormous sequencing capabilities of our times might be reaching the point of overflowing the possibilities to analyse data and allow for a feedback on where to focus the available resources. We have now a foreseeable future in which most bacterial species will have an annotated genome. However, we know also that most prokaryotic diversity would not be included there. On the one hand, there is the problem of many groups not being easily amenable to culture and hence not represented in culture-centred microbial taxonomy. On the other hand, the gene pools present in one species can be orders of magnitude larger than the genome of one strain (selected for genome sequencing). Contrasting with eukaryotic genomes, the repertoire of genes present in one prokaryotic cell genome does not correlate stringently with its taxonomic identity. Hence gene catalogues from one environment might provide more meaningful information than the classical species catalogues. Metagenomics or microbial environmental genomics provide a different tool that gravitates around the habitat rather than the species. Such a tool could be just the right way to complement "organismal genomics". Its potential to advance our understanding of microbial ecology and prokaryotic diversity and evolution is discussed.     

The PhyloCode: a critical discussion of its theoretical foundation

The definition of taxon names as formalized by the PhyloCode is based on Kripke's thesis of "rigid designation" that applies to Millian proper names. Accepting the thesis of "rigid designation" into systematics in turn is based on the thesis that species, and taxa, are individuals. These largely semantic and metaphysical issues are here contrasted with an epistemological approach to taxonomy. It is shown that the thesis of "rigid designation" if deployed in taxonomy introduces a new essentialism into systematics, which is exactly what the PhyloCode was designed to avoid. Rigidly designating names are not supposed to change their meaning, but if the shifting constitution of a clade is thought to cause a shift of meaning of the taxon name, then the taxon name is not a "rigid designator". Phylogenetic nomenclature either fails to preserve the stability of meaning of taxon names that it propagates, or it is rendered inconsistent with its own philosophical background. The alternative explored here is to conceptualize taxa as natural kinds, and to replace the analytic definition of taxon names by their explanatory definition. Such conceptualization of taxa allows taxon names to better track the results of ongoing empirical research. The semantic as well as epistemic gain is that if taxon names are associated with natural kind terms instead of being proper names, the composition of the taxon will naturally determine the meaning of its name.
© The Willi Hennig Society 2006.     

Microbial species delineation using whole genome sequences

Increased sequencing of microbial genomes has revealed that prevailing prokaryotic species assignments can be inconsistent with whole genome information for a significant number of species. The long-standing need for a systematic and scalable species assignment technique can be met by the genome-wide Average Nucleotide Identity (gANI) metric, which is widely acknowledged as a robust measure of genomic relatedness. In this work, we demonstrate that the combination of gANI and the alignment fraction (AF) between two genomes accurately reflects their genomic relatedness. We introduce an efficient implementation of AF,gANI and discuss its successful application to 86.5M genome pairs between 13,151 prokaryotic genomes assigned to 3032 species. Subsequently, by comparing the genome clusters obtained from complete linkage clustering of these pairs to existing taxonomy, we observed that nearly 18% of all prokaryotic species suffer from anomalies in species definition. Our results can be used to explore central questions such as whether microorganisms form a continuum of genetic diversity or distinct species represented by distinct genetic signatures. We propose that this precise and objective AF,gANI-based species definition: the MiSI (Microbial Species Identifier) method, be used to address previous inconsistencies in species classification and as the primary guide for new taxonomic species assignment, supplemented by the traditional polyphasic approach, as required.     

Phylogenetic definitions and taxonomic philosophy

An examination of the post-Darwinian history of biological taxonomy reveals an implicit assumption that the definitions of taxon names consist of lists of organismal traits. That assumption represents a failure to grant the concept of evolution a central role in taxonomy, and it causes conflicts between traditional methods of defining taxon names and evolutionary concepts of taxa. Phylogenetic definitions of taxon names (de Queiroz and Gauthier 1990) grant the concept of common ancestry a central role in the definitions of taxon names and thus constitute an important step in the development of phylogenetic taxonomy. By treating phylogenetic relationships rather than organismal traits as necessary and sufficient properties, phylogenetic definitions remove conflicts between the definitions of taxon names and evolutionary concepts of taxa. The general method of definition represented by phylogenetic definitions of clade names can be applied to the names of other kinds of composite wholes, including populations and biological species. That the names of individuals (composite wholes) can be defined in terms of necessary and sufficient properties provides the foundation for a synthesis of seemingly incompatible positions held by contemporary individualists and essentialists concerning the nature of taxa and the definitions of taxon names.     

Towards a genome-based taxonomy for prokaryotes

The ranks higher than the species in the prokaryotic taxonomy are primarily designated based on phylogenetic analysis of the 16S rRNA gene sequences, but no definite standards exist for the absolute relatedness (measured by 16S rRNA or other means) between the ranks. Accordingly, it remains unknown how comparable the ranks are between different organisms. To gain insights into this question, we studied the relationship between shared gene content and genetic relatedness for 175 fully sequenced strains, using as a robust measure of relatedness the average amino acid identity (AAI) of the shared genes. Our results reveal that adjacent ranks (e.g., phylum versus class) frequently show extensive overlap in terms of genetic and gene content relatedness of the grouped organisms, and hence, the current system is of limited predictive power in this respect. The overlap between nonadjacent ranks (e.g., phylum versus family) is generally limited and attributable to clear inconsistencies of the taxonomy. In addition to providing means for standardizing taxonomy, our AAI-based approach provides a means to evaluate the robustness of alternative genetic markers for phylogenetic purposes. For instance, the 23S rRNA gene was found to be as good a marker as the 16S rRNA gene, while several of the widely distributed protein-coding genes, such as the RNA polymerase and gyrase subunits, show a strong phylogenetic signal, albeit less strong than the rRNA genes (0.78 > R2 > 0.69 for the protein-coding genes versus R2 = 0.84 for the rRNA genes). The AAI approach outlined here could contribute significantly to a genome-based taxonomy for all microbial organisms.     

我国原核微生物分类学七十年北大核心

2022年和2023年分别是中国微生物学会成立70周年和《微生物学报》创刊70周年,我国原核微生物分类学研究从白手起家、跟踪模仿到逐步走上国际舞台,从跟跑、并跑到整体处于国际先进行列,部分领域领跑国际同行,并为国际同行提供微生物数据服务,也已走过了70个岁月。这些成绩的取得是我国原核微生物分类领域学者几代人努力的结果,是与国家发展、民族复兴同频共振的结果。特别是近30年,我国原核微生物分类无论从理论、方法创新还是新物种发现方面都取得了令人瞩目的成绩,在国际上的地位和影响力日益提高,已经逐渐成为国际原核系统分类领域的主导力量。本文本着尊重历史、客观求实的态度梳理了70年来我国在细菌和古菌分类学及相关领域的发展脉络和取得的成绩,并结合该领域最新发展方向进行了展望。     

Dialogue on the nomenclature and classification of prokaryotes

The application of next generation sequencing and molecular ecology to the systematics and taxonomy of prokaryotes offers enormous insights into prokaryotic biology. This discussion explores some major disagreements but also considers the opportunities associated with the nomenclature of the uncultured taxa, the use of genome sequences as type material, the plurality of the nomenclatural code, and the roles of an official or computer-assisted taxonomy.     

The bacterial species dilemma and the genomic-phylogenetic species concept

The number of species of Bacteria and Archaea (ca 5000) is surprisingly small considering their early evolution, genetic diversity and residence in all ecosystems. The bacterial species definition accounts in part for the small number of named species. The primary procedures required to identify new species of Bacteria and Archaea are DNA-DNA hybridization and phenotypic characterization. Recently, 16S rRNA gene sequencing and phylogenetic analysis have been applied to bacterial taxonomy. Although 16S phylogeny is arguably excellent for classification of Bacteria and Archaea from the Domain level down to the family or genus, it lacks resolution below that level. Newer approaches, including multilocus sequence analysis, and genome sequence and microarray analyses, promise to provide necessary information to better understand bacterial speciation. Indeed, recent data using these approaches, while meagre, support the view that speciation processes may occur at the subspecies level within ecological niches (ecovars) and owing to biogeography (geovars). A major dilemma for bacterial taxonomists is how to incorporate this new information into the present hierarchical system for classification of Bacteria and Archaea without causing undesirable confusion and contention. This author proposes the genomic-phylogenetic species concept (GPSC) for the taxonomy of prokaryotes. The aim is twofold. First, the GPSC would provide a conceptual and testable framework for bacterial taxonomy. Second, the GPSC would replace the burdensome requirement for DNA hybridization presently needed to describe new species. Furthermore, the GPSC is consistent with the present treatment at higher taxonomic levels.     

Prokaryotic taxonomy and phylogeny in the genomic era: advancements and challenges ahead

Advancing prokaryotic taxonomy constitutes a contemporary academic challenge as well as practical necessity. Genome sequencing has greatly facilitated the evaluation of the current taxonomic system and the development of simpler, more portable and accurate, sequence-based alternatives to substitute for the traditional cumbersome methods. Studies based on the former genome-enabled methods reveal that existing taxonomic designations, including the species level, correspond frequently to a continuum of genetic diversity as opposed to natural groupings (e.g. biological species). Improving such artificial and often ambiguous taxonomic designations, however, will require larger genomic datasets and more carefully designed sampling of natural populations. Only then can the promise for a superior genome-based taxonomy materialize.     

Quantifying spatial classification uncertainties of the historical Wisconsin landscape (USA)

Landscape feature can be classified by creating categories based on aggregation of spatially explicit information. However, many landscape features appear continuous rather than discrete. The aggregation process likely involves loss of information and introduces a variety of uncertainties whose degree and extent may differ spatially. Since landscape classifications have found wide application in e.g. natural resource policies or ecological research, assessments of spatial classification uncertainties are required.
We present a quantitative framework to identify the degree of landscape continuity (fuzziness) and structure (categorization) based on fuzzy classification and offer measures to quantify uncertainties originating from aggregating features into categories. Fuzzy classification is a non-hierarchical, quantitative method of assessing class definitions using degrees of association between features and class. This results in classes which are well defined and compositionally distinct, as well as classes which are less clearly defined but which, to various degrees, share characteristics with some or all classes. The spatial variation in the degree of class definition on the landscape is used to assess classification uncertainties. The two aspects of uncertainty investigated are the degree of association of a feature with the overall class definitions (membership diffusion), and the class-specific degree of association of each pixel on the landscape with each class (membership saturation).
Three classification scenarios, one fuzzy and one discrete, of the historical landscape of Wisconsin (USA) were compared for spatial classification uncertainties. Membership diffusion is highest in topographically heterogeneous environments, or areas characterized by many species occupying similar ecological niches. Classification uncertainties for individual classes show that differentiated species distributions can be identified, not only distribution centers.     

Contributions of molecular phylogenetics to foraminiferal taxonomy: General overview and example of Pseudoeponides falsobeccarii

Molecular phylogenetics gives new insights into the taxonomy of foraminifera, independent of their morphology. After a survey of the present knowledge on how molecular phylogeny can contribute to foraminiferal taxonomy, we present an applied example. The comparison of ribosomal DNA (rDNA) sequences belonging to the SSU (Small Subunit) and LSU (Large Subunit) genes of Pseudoeponides falsobeccarii with other similar sequences of rotaliids available in GenBank shows that this species actually belongs to the genus Ammonia, because it groups inside the other Ammonia sequences instead of forming a distinct clade. Moreover, Ammonia falsobeccarii forms a clade well separated from other Ammonia phylotypes, meaning that it can be considered as a distinct species, and not as an ecophenotype of one of the other Ammonia species.     

Numerical taxonomy: criticisms and critiques: Presidential address to the Systematics Association, November 1970

Some recent criticisms and critiques of numerical taxonomy are reviewed, together with some of its present shortcomings. It is pointed out that most of the problems are equally severe for orthodox taxonomy, and many of them can only be investigated by numerical techniques. The reasons for the general success of numerical methods in bacterial classification are discussed. Besides bringing deeper insights into taxonomy as a whole, numerical taxonomy is entering a new and heuristic phase, which includes potential applications to the study of evolution.     

Digest: Propagule pressure might not matter in the establishment of invasive prokaryotes*

Lavigne et al. developed models to investigate the adaptive colonization of sink environments by asexual organisms. Their results have clear relevance to the spread of infectious disease, but they may also provide insights into prokaryotic invasions into natural communities. Their results show that propagule pressure might not be a good predictor of invasion success in prokaryotes, suggesting that more work is needed to understand how microbial invasions differ from those of plants and animals.     

Extinction models and conservation

In their seminal book, The Theory of Island Biogeography, MacArthur and Wilson (1) proposed that '(the principles derived from island biogeography] apply, and will apply to an accelerating extent in the future, to formerly continuous natural habitats now being broken up by the encroachment of civilization'. Debate over the validity of this statement has raged for a decade, focusing particularly on reserve design 'rules' and whether a single large reserve or several small reserves will protect more species? These arguments have led to some new insights, if not a fresh perspective, on some old but fundamental topics in ecology and evolution. Most notable among these is extinction.     

An integrative taxonomic revision of the Tarentola geckos (Squamata,Phyllodactylidae) of the Cape Verde Islands

Recent phylogeographical analyses using mitochondrial DNA (mtDNA) sequences indicate that the Tarentola geckos from the Cape Verde archipelago originated from a propagule that dispersed from the Canary Islands approximately 7.7 Mya and that underwent a fast evolutionary radiation. Molecular analyses carried out to date clearly show some incongruences with the current taxonomy of Tarentola from the Cape Verde Islands, with some species being paraphyletic or polyphyletic, and several independently evolving lineages needing formal taxonomic recognition. The aim of this study was to clarify the systematics of this group to unravel its taxonomy by applying an integrative approach based on information from three independent sources: mtDNA, nuclear genes, and morphology. As a result of this taxonomic revision, two novel species for the islands of S. Nicolau and Fogo are described and eight subspecies are upgraded to species level. Moreover, an identification key for the genus Tarentola from the Cape Verde archipelago is presented. This study reconciles taxonomy and phylogeny in this group, provides a better understanding of diversity patterns, new insights on evolutionary hypotheses, and supports the basic framework for the future management and conservation of this unique reptile radiation. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 328–360.     

The tree, the network, and the species

To enrich the Hennigian internodal conception of species, a new formalization of the definition of the species concept is proposed. This rigorous definition allows for considerable unification of the various, and sometimes conflicting, techniques of species delimitation used in practice. First, the domain of such a definition is set out, namely, the set of all organisms on Earth, past, present, and future. Next, the focus is on the genealogical relationship among organisms, which provides the key to analysing the giant or global genealogical network (GGN) connecting all these organisms. This leads to the construction of an algorithm revealing the topological structure of the GGN, from families to lineages, ending up with a definition of species as equivalence classes of organisms corresponding to branches of the 'tree of life'. Such a theoretical definition of the species concept must be accompanied by various recognition criteria to be operational. These criteria are, for example, the ill-named 'biological species concepts', 'phylogenetic species concepts', etc., usually, but wrongly, presented as definitions of the species concept. Besides clarifying this disputed point, the definition in the present study displays the huge diversity of the scales (time-scale and population size) involved in actual species, thus explaining away the classical problems raised by previous attempts at defining the species concept (uniparental reproduction, temporal depth of species, and hybridization).  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 509–521.