A systematics for discovering the fundamental units of bacterial diversity

Bacterial systematists face unique challenges when trying to identify ecologically meaningful units of biological diversity. Whereas plant and animal systematists are guided by a theory-based concept of species, microbiologists have yet to agree upon a set of ecological and evolutionary properties that will serve to define a bacterial species. Advances in molecular techniques have given us a glimpse of the tremendous diversity present within the microbial world, but significant work remains to be done in order to understand the ecological and evolutionary dynamics that can account for the origin, maintenance, and distribution of that diversity. We have developed a conceptual framework that uses ecological and evolutionary theory to identify the DNA sequence clusters most likely corresponding to the fundamental units of bacterial diversity. Taking into account diverse models of bacterial evolution, we argue that bacterial systematics should seek to identify ecologically distinct groups with evidence of a history of coexistence, as based on interpretation of sequence clusters. This would establish a theory-based species unit that holds the dynamic properties broadly attributed to species outside of microbiology.     

Influence of plant developmental stage on microbial community structure and activity in the rhizosphere of three field crops

Seasonal shifts in rhizosphere microbial populations were investigated to follow the influence of plant developmental stage. A field study of indigenous microbial rhizosphere communities was undertaken on pea (Pisum satvium var. quincy), wheat (Triticum aestivum var. pena wawa) and sugar beet (Beta vulgaris var. amythyst). Rhizosphere community diversity and substrate utilization patterns were followed throughout a growing season, by culturing, rRNA gene density gradient gel electrophoresis and BIOLOG. Culturable bacterial and fungal rhizosphere community densities were stable in pea and wheat rhizospheres, with dynamic shifts observed in the sugar beet rhizosphere. Successional shifts in bacterial and fungal diversity as plants mature demonstrated that different plants select and define their own functional rhizosphere communities. Assessment of metabolic activity and resource utilization by bacterial community-level physiological profiling demonstrated greater similarities between different plant species rhizosphere communities at the same than at different developmental stages. Marked temporal shifts in diversity and relative activity were observed in rhizosphere bacterial communities with developmental stage for all plant species studied. Shifts in the diversity of fungal and bacterial communities were more pronounced in maturing pea and sugar beet plants. This detailed study demonstrates that plant species select for specialized microbial communities that change in response to plant growth and plant inputs.     

Use of quantum dot luminescent probes to achieve single-cell resolution of human oral bacteria in biofilms

Oral biofilms are multispecies communities, and in their nascent stages of development, numerous bacterial species engage in interspecies interactions. Better insight into the spatial relationship between different species and how species diversity increases over time can guide our understanding of the role of interspecies interactions in the development of the biofilms. Quantum dots (QD) are semiconductor nanocrystals and have emerged as a promising tool for labeling and detection of bacteria. We sought to apply QD-based primary immunofluorescence for labeling of bacterial cells with in vitro and in vivo biofilms and to compare this approach with the fluorophore-based primary immunofluorescence approach we have used previously. To investigate QD-based primary immunofluorescence as the means to detect distinct targets with single-cell resolution, we conjugated polyclonal and monoclonal antibodies to the QD surface. We also conducted simultaneous QD conjugate-based and fluorophore conjugate-based immunofluorescence and showed that these conjugates were complementary tools in immunofluorescence applications. Planktonic and biofilm cells were labeled effectively by considering two factors: the final nanomolar concentration of QD conjugate and the amount of antibody conjugated to the QD, which we define as the degree of labeling. These advances in the application of QD-based immunofluorescence for the study of biofilms in vitro and in vivo will help to define bacterial community architecture and to facilitate investigations of interactions between bacterial species in these communities.     

Use of Quantum Dot Luminescent Probes To Achieve Single-Cell Resolution of Human Oral Bacteria in Biofilms

Oral biofilms are multispecies communities, and in their nascent stages of development, numerous bacterial species engage in interspecies interactions. Better insight into the spatial relationship between different species and how species diversity increases over time can guide our understanding of the role of interspecies interactions in the development of the biofilms. Quantum dots (QD) are semiconductor nanocrystals and have emerged as a promising tool for labeling and detection of bacteria. We sought to apply QD-based primary immunofluorescence for labeling of bacterial cells with in vitro and in vivo biofilms and to compare this approach with the fluorophore-based primary immunofluorescence approach we have used previously. To investigate QD-based primary immunofluorescence as the means to detect distinct targets with single-cell resolution, we conjugated polyclonal and monoclonal antibodies to the QD surface. We also conducted simultaneous QD conjugate-based and fluorophore conjugate-based immunofluorescence and showed that these conjugates were complementary tools in immunofluorescence applications. Planktonic and biofilm cells were labeled effectively by considering two factors: the final nanomolar concentration of QD conjugate and the amount of antibody conjugated to the QD, which we define as the degree of labeling. These advances in the application of QD-based immunofluorescence for the study of biofilms in vitro and in vivo will help to define bacterial community architecture and to facilitate investigations of interactions between bacterial species in these communities.     

Tree diversity and functional leaf traits drive herbivore‐associated microbiomes in subtropical China

Herbivorous insects acquire microorganisms from host plants or soil, but it remains unclear how the diversity and functional composition of host plants contribute to structuring herbivore microbiomes. Within a controlled tree diversity setting, we used DNA metabarcoding of 16S rRNA to assess the contribution of Lepidoptera species and their local environment (particularly, tree diversity, host tree species, and leaf traits) to the composition of associated bacterial communities. In total, we obtained 7,909 bacterial OTUs from 634 caterpillar individuals comprising 146 species. Tree diversity was found to drive the diversity of caterpillar‐associated bacteria both directly and indirectly via effects on caterpillar communities, and tree diversity was a stronger predictor of bacterial diversity than diversity of caterpillars. Leaf toughness and dry matter content were important traits of the host plant determining bacterial species composition, while leaf calcium and potassium concentration influenced bacterial richness. Our study reveals previously unknown linkages between trees and their characteristics, herbivore insects, and their associated microbes, which contributes to developing a more nuanced understanding of functional dependencies between herbivores and their environment, and has implications for the consequences of plant diversity loss for trophic interactions.     

Differences in vegetation composition and plant species identity lead to only minor changes in soil-borne microbial communities in a former arable field

To examine the relationship between plant species composition and microbial community diversity and structure, we carried out a molecular analysis of microbial community structure and diversity in two field experiments. In the first experiment, we examined bacterial community structure in bulk and rhizosphere soils in fields exposed to different plant diversity treatments, via a 16S rRNA gene clone library approach. Clear differences were observed between bacterial communities of the bulk soil and the rhizosphere, with the latter containing lower bacterial diversity. The second experiment focused on the influence of 12 different native grassland plant species on bacterial community size and structure in the rhizosphere, as well as the structure of Acidobacteria and Verrucomicrobia community structures. In general, bacterial and phylum-specific quantitative PCR and PCR-denaturing gradient gel electrophoresis revealed only weak influences of plant species on rhizosphere communities. Thus, although plants did exert an influence on microbial species composition and diversity, these interactions were not specific and selective enough to lead to major impacts of vegetation composition and plant species on below-ground microbial communities.     

Bacterial species and evolution: Theoretical and practical perspectives

A discussion of the species problem in modern evolutionary biology serves as the point of departure for an exploration of how the basic science aspects of this problem relate to efforts to map bacterial diversity for practical pursuits—for prospecting among the bacteria for useful genes and gene-products. Out of a confusing array of species concepts, the Cohesion Species Concept seems the most appropriate and useful for analyzing bacterial diversity. Techniques of allozyme analysis and DNA fingerprinting can be used to put this concept into practice to map bacterial genetic diversity, though the concept requires minor modification to encompass cases of complete asexuality. Examples from studies of phenetically definedBacillus species provide very partial maps of genetic population structure. A major conclusion is that such maps frequently reveal deep genetic subdivision within the phenetically defined specles; divisions that in some cases are clearly distinct genetic species. Knowledge of such subdivisions is bound to make prospecting within bacterial diversity more effective. Under the general concept of genetic cohesion a hypothetical framework for thinking about the full range of species conditions that might exist among bacteria is developed and the consequences of each such model for species delineation, and species identification are discussed. Modes of bacterial evolution, and a theory of bacterial speciation with and without genetic recombination, are examined. The essay concludes with thoughts about prospects for very extensive mapping of bacterial diversity in the service of future efforts to find useful products. In this context, evolutionary biology becomes the handmaiden of important industrial activities. A few examples of past success in commercializing bacterial gene-products from species ofBacillus and a few other bacteria are reviewed.     

Does adaptive radiation of a host lineage promote ecological diversity of its bacterial communities? A test using gut microbiota of Anolis lizards

Adaptive radiations provide unique opportunities to test whether and how recent ecological and evolutionary diversification of host species structures the composition of entire bacterial communities. We used 16S rRNA gene sequencing of faecal samples to test for differences in the gut microbiota of six species of Puerto Rican Anolis lizards characterized by the evolution of distinct ‘ecomorphs’ related to differences in habitat use. We found substantial variation in the composition of the microbiota within each species and ecomorph (trunk‐crown, trunk‐ground, grass‐bush), but no differences in bacterial alpha diversity among species or ecomorphs. Beta diversity analyses revealed subtle but significant differences in bacterial composition related to host phylogeny and species, but these differences were not consistently associated with Anolis ecomorph. Comparison of a trunk‐ground species from this clade (A. cristatellus) with a distantly related member of the same ecomorph class (A. sagrei) where the two species have been introduced and are now sympatric in Florida revealed pronounced differences in the alpha diversity and beta diversity of their microbiota despite their ecological similarity. Comparisons of these populations with allopatric conspecifics also revealed geographic differences in bacterial alpha diversity and beta diversity within each species. Finally, we observed high intraindividual variation over time and strong effects of a simplified laboratory diet on the microbiota of A. sagrei. Collectively, our results indicate that bacterial communities are only weakly shaped by the diversification of their lizard hosts due to the strikingly high levels of bacterial diversity and variation observed within Anolis species.     

Genetic difference but functional similarity among fish gut bacterial communities through molecular and biochemical fingerprints

Considering the major involvement of gut microflora in the digestive function of various macro-organisms, bacterial communities inhabiting fish guts may be the main actors of organic matter degradation by fish. Nevertheless, the extent and the sources of variability in the degradation potential of gut bacterial communities are largely overlooked. Using Biolog Ecoplate? and denaturing gradient gel electrophoresis (DGGE), we explored functional (i.e. the ability to degrade organic matter) and genetic (i.e. identification of DGGE banding patterns) diversity of fish gut bacterial communities, respectively. Gut bacterial communities were extracted from fish species characterized by different diets sampled along a salinity gradient in the Patos-Mirim lagoons complex (Brazil). We found that functional diversity was surprisingly unrelated to genetic diversity of gut bacterial communities. Functional diversity was not affected by the sampling site but by fish species and diet, whereas genetic diversity was significantly influenced by all three factors. Overall, the functional diversity was consistently high across fish individuals and species, suggesting a wide functional niche breadth and a high potential of organic matter degradation. We conclude that fish gut bacterial communities may strongly contribute to nutrient cycling regardless of their genetic diversity and environment.     

Both rare and common species make unique contributions to functional diversity in an ecosystem unaffected by human activities

Aim

Rare species typically contribute more to functional diversity than common species. However, humans have altered the occupancy and abundance patterns of many species—the basis upon which we define “rarity.” Here, we use a globally unique dataset from hydrothermal vents—an untouched ecosystem—to test whether rare species over‐contribute to functional diversity.

Location

Juan de Fuca Ridge hydrothermal vent fields, Northeast Pacific Ocean.

Methods

We first conduct a comprehensive review to set up expectations for the relative contributions of rare and common species to functional diversity. We then quantify the rarity and commonness of 37 vent species with relevant trait information to assess the relationship between rarity and functional distinctiveness—a measure of the uniqueness of the traits of a species relative to traits of coexisting species. Next, we randomly assemble communities to test whether rare species over‐contribute to functional diversity in artificial assemblages ranging in species richness. Then, we test whether biotic interactions influence functional diversity contributions by comparing the observed contribution of each species to a null expectation. Finally, we identify traits driving functional distinctiveness using a distance‐based redundancy analysis.

Results

Across functional diversity metrics and species richness levels, we find that both rare and common species can contribute functional uniqueness. Some species always offer unique trait combinations, and these species host bacterial symbionts and provide habitat complexity. Moreover, we find that contributions of species to functional diversity may be influenced by biotic interactions.

Main conclusions

Our findings show that many common species make persistent, unique contributions to functional diversity. Thus, it is key to consider whether the abundance and occupancy of species have been reduced, relative to historical baselines, when interpreting the contributions of rare species to functional diversity. Our work highlights the importance of testing ecological theory in ecosystems unaffected by human activities for the conservation of biodiversity.     

Density compensation can cause no effect of biodiversity on ecosystem functioning

The role of density compensation (the decline of species density with increasing diversity), in the context of biodiversity and ecosystem functioning, has not been explicitly explored. I used aquatic microbial communities containing bacterivorous consumers (protozoans and rotifers) to investigate whether competition can lead to density compensation and whether density compensation can contribute to the relationship between biodiversity and ecosystem functioning. The experiment employed a nested design in which the consumer diversity gradient (0, 1, 2 or 4 species) was constructed by drawing all possible species or species combinations at each diversity level from a five-species pool. All consumer species coexisted but there was little evidence for overyielding or species dominance, suggesting weak complementarity and sampling effects. Rather, increasing number of consumer species resulted in community-wide density compensation, such that aggregate consumer biomass was unaffected by consumer diversity. Whereas culturable bacterial density declined as consumer diversity increased, total bacterial density showed no discernible response to changes in consumer diversity, a result probably due in part to heterogeneity in bacterial edibility. This study demonstrates the potential for density compensation to shape the relationship between biodiversity and ecosystem functioning.     

Dominant plant species shape soil bacterial community in semiarid sandy land of northern China

Plant species affect soil bacterial diversity and compositions. However, little is known about the role of dominant plant species in shaping the soil bacterial community during the restoration of sandy grasslands in Horqin Sandy Land, northern China. We established a mesocosm pots experiment to investigate short‐term responses of soil bacterial diversity and composition, and the related soil properties in degraded soils without vegetation (bare sand as the control, CK) to restoration with five plant species that dominate across restoration stages: Agriophyllum squarrosum (AS), Artemisia halodendron (AH), Setaria viridis (SV), Chenopodium acuminatum (CA), and Corispermum macrocarpum (CM). We used redundancy analysis (RDA) to analyze the association between soil bacterial composition and soil properties in different plant species. Our results indicated that soil bacterial diversity was significantly lower in vegetated soils independent of plant species than in the CK. Specifically, soil bacterial species richness and diversity were lower under the shrub AH and the herbaceous plants AS, SV, and CA, and soil bacterial abundance was lower under AH compared with the CK. A field investigation confirmed the same trends where soil bacteria diversity was lower under AS and AH than in bare sand. The high‐sequence annotation analysis showed that Proteobacteria, Actinobacteria, and Bacteroidetes were the most common phyla in sandy land irrespective of soil plant cover. The OTUs (operational taxonomic units) indicated that some bacterial species were specific to the host plants. Relative to bare sand (CK), soils with vegetative cover exhibited lower soil water content and temperature, and higher soil carbon and nitrogen contents. The RDA result indicated that, in addition to plant species, soil water and nitrogen contents were the most important factors shaping soil bacterial composition in semiarid sandy land. Our study from the pot and field investigations clearly demonstrated that planting dominant species in bare sand impacts bacterial diversity. In semiarid ecosystems, changes in the dominant plant species during vegetation restoration efforts can affect the soil bacterial diversity and composition through the direct effects of plants and the indirect effects of soil properties that are driven by plant species.     

Bacterial evolution: chromosome arithmetic and geometry

Recent sequencing projects have characterized bacterial genomes that are organized onto elements of various sizes, shapes and numbers. Aside from its biological relevance and curiosity, this diversity calls into question the way that we define bacterial chromosomes.     

Santa Rosalia revisited: Why are there so many species of bacteria?

The diversity of bacteria in the world is very poorly known. Usually less than one percent of the bacteria from natural communities can be grown in the laboratory. This has caused us to underestimate bacterial diversity and biased our view of bacterial communities. The tools are now available to estimate the number of bacterial species in a community and to estimate the difference between communities. Using what data are available, I have estimated that thirty grams of forest soil contains over half a million species. The species difference between related communities suggests that the number of species of bacteria may be more than a thousand million. I suppose that the explanation for such a large number of bacterial species is simply that speciation in bacteria is easy and extinction difficult, giving a rate of speciation higher than the rate of extinction, leading to an ever increasing number of species over time. The idea that speciation is easy is justified from the results of recent experimental work in bacterial evolution.     

Consumer vs environmental productivity control of bacterial diversity and bacteria-mediated organic matter decomposition

We experimentally investigated the determinants (consumer vs environmental productivity) and functional consequences of bacterial diversity using a simple laboratory bacterial assemblage and the bacterivorous ciliated protist Colpidium striatum . A two-way factorial design was used, with two productivity levels crossed with the presence/absence of Colpidium striatum . The effects of productivity and predation on bacterial diversity were similar to those previously reported for plant diversity in systems characterized by high productivities, with increased productivity reducing diversity and predation increasing diversity. When Colpidium was present, changes in trophic level biomass and individual bacterial species abundance in response to increased productivity were largely consistent with the keystone predation model. Importantly, Colpidium predation also resulted in an increase in particulate organic matter decomposition, largely due to an increase in the abundance of one single bacterial species that appeared resistant to predation. These results suggest that changes in community structure as a result of trophic interactions and other factors may have profound consequences for important ecosystem functions. We suggest that future research should take an integrative approach and study causes and consequences of biodiversity simultaneously.     

Effects of maternal genotypic identity and genetic diversity of the red mangrove Rhizophora mangle on associated soil bacterial communities: A field‐based experiment

Loss of plant biodiversity can result in reduced abundance and diversity of associated species with implications for ecosystem functioning. In ecosystems low in plant species diversity, such as Neotropical mangrove forests, it is thought that genetic diversity within the dominant plant species could play an important role in shaping associated communities. Here, we used a manipulative field experiment to study the effects of maternal genotypic identity and genetic diversity of the red mangrove Rhizophora mangle on the composition and richness of associated soil bacterial communities. Using terminal restriction fragment length polymorphism (T‐RFLP) community fingerprinting, we found that bacterial community composition differed among R. mangle maternal genotypes but not with genetic diversity. Bacterial taxa richness, total soil nitrogen, and total soil carbon were not significantly affected by maternal genotypic identity or genetic diversity of R. mangle. Our findings show that genotype selection in reforestation projects could influence soil bacterial community composition. Further research is needed to determine what impact these bacterial community differences might have on ecosystem processes, such as carbon and nitrogen cycling.     

Effects of Different Re-vegetation Patterns on Soil Physicochemical Properties and Bacterial Community in Kunyang Phosphate-mine

Soil microorganisms are applied to evaluate soil quality and significant in restoration ecology for their important roles on nutrient cycle and sensitivity to environment changes. To investigate the effects of re vegetation pattern on soil physicochemical properties and soil bacterial community and the reasons for soil bacterial community discrepancy, the soils of three re vegetation patterns (grass of Miscanthus sinensis, the mixed forest of Alnus nepalensis, Cupressus torulosa and Quercus acutissima, the mixed forest of Cupressus torulosa and Alnus nepalensis) in Kunyang phosphate mine, near Kunming, Yunnan province of China were studied. Polymerase chain reaction denaturing gradient gel electrophoresis (PCR DGGE) and soil physicochemical indices were used to analyze soil bacterial communities (diversity and species composition) and soil physicochemical properties. The results indicated that re vegetation contributed to soil nutrients and soil physicochemical properties were different in these three patterns. As for soil bacterial community, the diversity and species composition varied in different patterns. Pearson correlation showed that soil bacterial community diversity was correlated with re vegetation pattern significantly but not with any soil physicochemical properties determined in this research. Soil bacterial species composition was strongly correlated with soil available nitrogen and re vegetation pattern but no others through Canonical Correspondence Analysis (CCA). It is concluded that both soil physicochemical properties and bacterial community (diversity and species composition) vary in different re vegetation patterns and the reason for diversity difference is re vegetation pattern while the affecting factors of soil bacterial community composition are soil available nitrogen and re vegetation pattern orderly.     

巨菌草不同生长时期对砒砂岩地区土壤理化性质及细菌群落结构的影响

[目的]揭示巨菌草种植前及其不同生长时期对砒砂岩地区土壤基本理化性质及细菌群落结构的影响,为更好应用巨菌草改良砒砂岩地区土壤提供理论依据。[方法]运用Illumina MiSeq高通量测序技术,在门、属水平上比较砒砂岩地区巨菌草不同生长时期对土壤细菌结构及多样性的影响。[结果]巨菌草种植后,三块样地的含水量在巨菌草分蘖期最低,在成熟期达到最高,土壤有机含量有所增加,土壤pH值变化不明显。细菌菌群结构：各样地的土壤细菌在巨菌草种植前及其生长的四个时期,门水平上的优势菌群均为放线菌门（Actinobacteria）、变形菌门（Proteobacteria）;在属水平上,含量最高的是未分类（unidentified）的细菌。Alpha多样性及群落组成丰度分析表明,三块样地种植巨菌草后,其细菌多样性和物种丰度均比种植前高,农田地土壤细菌多样性和物种丰度在巨菌草拔节期达到峰值,在分蘖期土壤细菌数量最大,比种植前土壤细菌数量高出8倍多,达2.09×10⁷ CFU/g干土,山顶地土壤细菌多样性和物种丰度在巨菌草分蘖期达到峰值,河滩地土壤细菌多样性在巨菌草苗期达到峰值,其丰富度在拔节期达到最大。[结论]砒砂岩地区土壤细菌群落结构在巨菌草不同生长时期相似,巨菌草能明显促进砒砂岩地区土壤细菌菌群多样性和丰富度,能有效提高土壤有机质含量,但砒砂岩地区各样地土壤细菌多样性、数量和有机质含量达到峰值的时期不同。     

Multilocus Sequence Analysis of Xanthomonads Causing Bacterial Spot of Tomato and Pepper Plants Reveals Strains Generated by Recombination among Species and Recent Global Spread of Xanthomonas gardneri

Four Xanthomonas species are known to cause bacterial spot of tomato and pepper, but the global distribution and genetic diversity of these species are not well understood. A collection of bacterial spot-causing strains from the Americas, Africa, Southeast Asia, and New Zealand were characterized for genetic diversity and phylogenetic relationships using multilocus sequence analysis of six housekeeping genes. By examining strains from different continents, we found unexpected phylogeographic patterns, including the global distribution of a single multilocus haplotype of X. gardneri, possible regional differentiation in X. vesicatoria, and high species diversity on tomato in Africa. In addition, we found evidence of multiple recombination events between X. euvesicatoria and X. perforans. Our results indicate that there have been shifts in the species composition of bacterial spot pathogen populations due to the global spread of dominant genotypes and that recombination between species has generated genetic diversity in these populations.     

树种多样性对土壤微生物群落结构和元素生物地球化学循环的影响研究进展

森林是陆地生态系统的重要组成部分,其巨大的生产力和生态服务功能对人类的生存和发展至关重要。森林树种多样性增加能够显著提高森林生产力,关于树种多样性如何影响地下生物多样性及生态功能逐渐受到国内外学者的广泛关注。从土壤微生物及其介导的元素生物地球化学循环这一视角出发,综述了树种多样性对土壤细菌和真菌多样性、群落结构及功能的影响,提出需要进一步深入研究的方向。总体来说,树种多样性有利于增加土壤细菌生物量和多样性,是预测病原性真菌和菌根真菌多样性及群落结构的重要生物因子。树种多样性能增加土壤有机碳储量,增强森林土壤的甲烷氧化能力,并提高土壤磷周转速率及有效磷含量。关于树种多样性对森林土壤氮循环的影响需考虑多样性假说和质量比假说的相对贡献。今后应加强树种多样性对多个营养级之间相互作用的研究;关注树种多样性对生态系统多功能的影响;加强学科交叉,引入微生物种群动态模型和气候模型等模型预测方法,研究树种多样性对全球气候变化的应对机制,以期促进地上植物多样性与地下生态系统功能关系的研究,增强森林生态系统应对未来全球环境变化的能力。