Long-Term Nitrogen Amendment Alters the Diversity and Assemblage of Soil Bacterial Communities in Tallgrass Prairie

Anthropogenic changes are altering the environmental conditions and the biota of ecosystems worldwide. In many temperate grasslands, such as North American tallgrass prairie, these changes include alteration in historically important disturbance regimes (e.g., frequency of fires) and enhanced availability of potentially limiting nutrients, particularly nitrogen. Such anthropogenically-driven changes in the environment are known to elicit substantial changes in plant and consumer communities aboveground, but much less is known about their effects on soil microbial communities. Due to the high diversity of soil microbes and methodological challenges associated with assessing microbial community composition, relatively few studies have addressed specific taxonomic changes underlying microbial community-level responses to different fire regimes or nutrient amendments in tallgrass prairie. We used deep sequencing of the V3 region of the 16S rRNA gene to explore the effects of contrasting fire regimes and nutrient enrichment on soil bacterial communities in a long-term (20 yrs) experiment in native tallgrass prairie in the eastern Central Plains. We focused on responses to nutrient amendments coupled with two extreme fire regimes (annual prescribed spring burning and complete fire exclusion). The dominant bacterial phyla identified were Proteobacteria, Verrucomicrobia, Bacteriodetes, Acidobacteria, Firmicutes, and Actinobacteria and made up 80% of all taxa quantified. Chronic nitrogen enrichment significantly impacted bacterial community diversity and community structure varied according to nitrogen treatment, but not phosphorus enrichment or fire regime. We also found significant responses of individual bacterial groups including Nitrospira and Gammaproteobacteria to long-term nitrogen enrichment. Our results show that soil nitrogen enrichment can significantly alter bacterial community diversity, structure, and individual taxa abundance, which have important implications for both managed and natural grassland ecosystems.     

Links between soil microbial communities and plant traits in a species‐rich grassland under long‐term climate change

Climate change can influence soil microorganisms directly by altering their growth and activity but also indirectly via effects on the vegetation, which modifies the availability of resources. Direct impacts of climate change on soil microorganisms can occur rapidly, whereas indirect effects mediated by shifts in plant community composition are not immediately apparent and likely to increase over time. We used molecular fingerprinting of bacterial and fungal communities in the soil to investigate the effects of 17 years of temperature and rainfall manipulations in a species‐rich grassland near Buxton, UK. We compared shifts in microbial community structure to changes in plant species composition and key plant traits across 78 microsites within plots subjected to winter heating, rainfall supplementation, or summer drought. We observed marked shifts in soil fungal and bacterial community structure in response to chronic summer drought. Importantly, although dominant microbial taxa were largely unaffected by drought, there were substantial changes in the abundances of subordinate fungal and bacterial taxa. In contrast to short‐term studies that report high resistance of soil fungi to drought, we observed substantial losses of fungal taxa in the summer drought treatments. There was moderate concordance between soil microbial communities and plant species composition within microsites. Vector fitting of community‐weighted mean plant traits to ordinations of soil bacterial and fungal communities showed that shifts in soil microbial community structure were related to plant traits representing the quality of resources available to soil microorganisms: the construction cost of leaf material, foliar carbon‐to‐nitrogen ratios, and leaf dry matter content. Thus, our study provides evidence that climate change could affect soil microbial communities indirectly via changes in plant inputs and highlights the importance of considering long‐term climate change effects, especially in nutrient‐poor systems with slow‐growing vegetation.     

Differential response of high-elevation planktonic bacterial community structure and metabolism to experimental nutrient enrichment

Nutrient enrichment of high-elevation freshwater ecosystems by atmospheric deposition is increasing worldwide, and bacteria are a key conduit for the metabolism of organic matter in these oligotrophic environments. We conducted two distinct in situ microcosm experiments in a high-elevation lake (Emerald Lake, Sierra Nevada, California, USA) to evaluate responses in bacterioplankton growth, carbon utilization, and community structure to short-term enrichment by nitrate and phosphate. The first experiment, conducted just following ice-off, employed dark dilution culture to directly assess the impact of nutrients on bacterioplankton growth and consumption of terrigenous dissolved organic matter during snowmelt. The second experiment, conducted in transparent microcosms during autumn overturn, examined how bacterioplankton in unmanipulated microbial communities responded to nutrients concomitant with increasing phytoplankton-derived organic matter. In both experiments, phosphate enrichment (but not nitrate) caused significant increases in bacterioplankton growth, changed particulate organic stoichiometry, and induced shifts in bacterial community composition, including consistent declines in the relative abundance of Actinobacteria. The dark dilution culture showed a significant increase in dissolved organic carbon removal in response to phosphate enrichment. In transparent microcosms nutrient enrichment had no effect on concentrations of chlorophyll, carbon, or the fluorescence characteristics of dissolved organic matter, suggesting that bacterioplankton responses were independent of phytoplankton responses. These results demonstrate that bacterioplankton communities in unproductive high-elevation habitats can rapidly alter their taxonomic composition and metabolism in response to short-term phosphate enrichment. Our results reinforce the key role that phosphorus plays in oligotrophic lake ecosystems, clarify the nature of bacterioplankton nutrient limitation, and emphasize that evaluation of eutrophication in these habitats should incorporate heterotrophic microbial communities and processes.     

Soil Water Content and Organic Carbon Availability Are Major Determinants of Soil Microbial Community Composition

Exploration of environmental factors governing soil microbial community composition is long overdue and now possible with improved methods for characterizing microbial communities. Previously, we observed that rice soil microbial communities were distinctly different from tomato soil microbial communities, despite management and seasonal variations within soil type. Potential contributing factors included types and amounts of organic inputs, organic carbon content, and timing and amounts of water inputs. Of these, both soil water content and organic carbon availability were highly correlated with observed differences in composition. We examined how organic carbon amendment (compost, vetch, or no amendment) and water additions (from air dry to flooded) affect microbial community composition. Using canonical correspondence analysis of phospholipid fatty acid data, we determined flooded, carbon-amended (+C) microcosm samples were distinctly different from other +C samples and unamended (–C) samples. Although flooding without organic carbon addition influenced composition some, organic carbon addition was necessary to substantially alter community composition. Organic carbon availability had the same general effects on microbial communities regardless of whether it was compost or vetch in origin. In addition, flooded samples, regardless of organic carbon inputs, had significantly lower ratios of fungal to bacterial biomarkers, whereas under drier conditions and increased organic carbon availability the microbial communities had higher proportions of fungal biomass. When comparing field and microcosm soil, flooded +C microcosm samples were most similar to field-collected rice soil, whereas all other treatments were more similar to field-collected tomato soil. Overall, manipulating water and carbon content selected for microbial communities similar to those observed when the same factors were manipulated at the field scale.     

How Does Conversion of Natural Tropical Rainforest Ecosystems Affect Soil Bacterial and Fungal Communities in the Nile River Watershed of Uganda?

Uganda''s forests are globally important for their conservation values but are under pressure from increasing human population and consumption. In this study, we examine how conversion of natural forest affects soil bacterial and fungal communities. Comparisons in paired natural forest and human-converted sites among four locations indicated that natural forest soils consistently had higher pH, organic carbon, nitrogen, and calcium, although variation among sites was large. Despite these differences, no effect on the diversity of dominant taxa for either bacterial or fungal communities was detected, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Composition of fungal communities did generally appear different in converted sites, but surprisingly, we did not observe a consistent pattern among sites. The spatial distribution of some taxa and community composition was associated with soil pH, organic carbon, phosphorus and sodium, suggesting that changes in soil communities were nuanced and require more robust metagenomic methods to understand the various components of the community. Given the close geographic proximity of the paired sampling sites, the similarity between natural and converted sites might be due to continued dispersal between treatments. Fungal communities showed greater environmental differentiation than bacterial communities, particularly according to soil pH. We detected biotic homogenization in converted ecosystems and substantial contribution of β-diversity to total diversity, indicating considerable geographic structure in soil biota in these forest communities. Overall, our results suggest that soil microbial communities are relatively resilient to forest conversion and despite a substantial and consistent change in the soil environment, the effects of conversion differed widely among sites. The substantial difference in soil chemistry, with generally lower nutrient quantity in converted sites, does bring into question, how long this resilience will last.     

The functional and taxonomic richness of wastewater treatment plant microbial communities are associated with each other and with ambient nitrogen and carbon availability

The number of functional traits of a wastewater treatment plant (WWTP) microbial community (i.e. functional richness) is thought to be an important determinant of its overall functional performance, but the ecological factors that determine functional richness remain unclear. The number of taxa within a community (i.e. taxonomic richness) is one ecological factor that might be important. Communities that contain more taxa are more likely to have more functional traits, and a positive association is therefore expected between functional and taxonomic richness. Empirical tests for this positive association among WWTP communities, however, are lacking. We address this knowledge gap by measuring the functional and taxonomic richness of 10 independent WWTP communities. We demonstrate that functional and taxonomic richness are positively associated with each other. We further demonstrate that functional and taxonomic richness are negatively associated with the effluent NH₄‐N and BOD₅ concentrations. This led us to hypothesize that correlated variation in functional and taxonomic richness is likely related to variation in ambient nitrogen and carbon availability. We finally demonstrate that this hypothesis is consistent with the functional and taxonomic attributes of the WWTP communities. Together, our results improve our basic understanding of the ecology and functioning of WWTP communities.     

海拔梯度变化对中亚热带黄山松土壤微生物生物量和群落结构的影响

全球变暖对陆地生态系统造成一系列生态问题,使这些问题将随着全球平均气温的升高而进一步加剧。海拔梯度变化是研究气候变暖对陆地生态系统影响的一种重要手段。目前为止利用海拔梯度对微生物影响的研究尚未定论,其主要原因是忽略了植被类型的影响。因此,以中亚热带戴云山的3个海拔(1300、1450、1600 m)的黄山松(Pinus taiwanensis)林为研究对象,探究沿海拔梯度的变化,森林土壤微生物生物量和微生物群落结构的响应变化。结果表明:土壤碳氮磷养分(SOC、TN、TP)、微生物生物量氮(MBN)、微生物生物量磷(MBP)和丛枝菌根真菌(AMF)、革兰氏阴性菌(GN)、真菌(Fungi)、总磷脂脂肪酸(T_(PLFA)),细菌∶真菌(F∶B)均随海拔升高显著下降,而革兰氏阳性菌∶革兰氏阴性菌(GP∶GN)随海拔升高呈相反的趋势。冗余分析(RDA)表明,温度(T)和可溶性有机氮(DON)是影响微生物群落结构的最重要的环境因子。研究表明:与1600 m海拔相比,1300 m海拔温度较高,土壤有机质矿化作用较强,土壤速效养分及微生物生物量随之增加,从而提高(Fungi)、细菌(Bacteria)等。因此,未来气候变暖将通过改变土壤碳氮磷养分来影响本区域微生物群落组成结构。这对进一步深入了解气候变化对山地生态系统土壤养分循环过程具有重要意义。     

A Coexisting Fungal-Bacterial Community Stabilizes Soil Decomposition Activity in a Microcosm Experiment

How diversity influences the stability of a community function is a major question in ecology. However, only limited empirical investigations of the diversity–stability relationship in soil microbial communities have been undertaken, despite the fundamental role of microbial communities in driving carbon and nutrient cycling in terrestrial ecosystems. In this study, we conducted a microcosm experiment to investigate the relationship between microbial diversity and stability of soil decomposition activities against changes in decomposition substrate quality by manipulating microbial community using selective biocides. We found that soil respiration rates and degradation enzyme activities by a coexisting fungal and bacterial community (a taxonomically diverse community) are more stable against changes in substrate quality (plant leaf materials) than those of a fungi-dominated or a bacteria-dominated community (less diverse community). Flexible changes in the microbial community composition and/or physiological state in the coexisting community against changes in substrate quality, as inferred by the soil lipid profile, may be the mechanism underlying this positive diversity–stability relationship. Our experiment demonstrated that the previously found positive diversity–stability relationship could also be valid in the soil microbial community. Our results also imply that the functional/taxonomic diversity and community ecology of soil microbes should be incorporated into the context of climate–ecosystem feedbacks. Changes in substrate quality, which could be induced by climate change, have impacts on decomposition process and carbon dioxide emission from soils, but such impacts may be attenuated by the functional diversity of soil microbial communities.     

Thirty-seven years of soil nitrogen and phosphorus fertility management shapes the structure and function of the soil microbial community in a Brown Chernozem

We tested whether levels of soil available nitrogen (N) and phosphorus (P) control the composition and function of the soil microbial community in a Brown Chernozemic soil on the Canadian Prairie. Soil dissolved organic carbon, N and P, and microbial communities structure (phospholipid fatty acid profile) and function (enzyme activity) were evaluated in the fallow and first wheat (Triticum aestivum L. cv. AC Eatonia) phases of fallow-wheat-wheat rotations where the wheat received soil test recommended rates of mineral N and P fertilizers (+N+P), or where N (?N+P) or P (+N?P) fertilizer use was withheld for 37 years. Differential fertilization modified soil N and P availability, and microbial community structure. Low N level was a major constraint when a rapidly growing wheat crop (heading stage) was drawing on the resource, reducing both plant N uptake and soil microbial biomass-C in ?N+P soils. Available P level in +N?P soils was about half that measured in P-fertilized soils, but P did not limit plant productivity or microbial development at that time. Changes in the microbial community structure seemingly buffered the impact of lower P availability in +N?P soils. Phosphatase activity was not involved, but increased abundance of arbuscular mycorrhizal fungi might be associated with this effect. Low soil N availability explained lower specific denitrification and higher specific nitrogenase activities in ?N+P soil growing wheat. Higher denitrification activity in +N+P soil could be attributed to higher soil C level and fertilization-induced shifts observed in the structure of the soil microbial community. Irrespective of the fertility level of the soil, all microbial communities grew at the relative growth rate of 17% day^?1 in a nutrient limitation assay that revealed no C, N or P limitation in these communities. We conclude that mineral fertilization, which modifies soil available N and P fertility, can be a selective force causing structural and functional shifts in the soil microbial community with a resulting impact on soil quality and nutrient fluxes.     

Rock structure drives the taxonomic and functional diversity of endolithic microbial communities in extreme environments

Endolithic (rock-dwelling) microbial communities are ubiquitous in hyper-arid deserts around the world and the last resort for life under extreme aridity. These communities are excellent models to explore biotic and abiotic drivers of diversity because they are of low complexity. Using high-throughput amplicon and metagenome sequencing, combined with X-ray computed tomography, we investigated how water availability and substrate architecture modulated the taxonomic and functional composition of gypsum endolithic communities in the Atacama Desert, Chile. We found that communities inhabiting gypsum rocks with a more fragmented substrate architecture had higher taxonomic and functional diversity, despite having less water available. This effect was tightly linked with community connectedness and likely the result of niche differentiation. Gypsum communities were functionally similar, yet adapted to their unique micro-habitats by modulating their carbon and energy acquisition strategies and their growth modalities. Reconstructed population genomes showed that these endolithic microbial populations encoded potential pathways for anoxygenic phototrophy and atmospheric hydrogen oxidation as supplemental energy sources.     

Microbial nitrogen and phosphorus co-limitation across permafrost region

The status of plant and microbial nutrient limitation have profound impacts on ecosystem carbon cycle in permafrost areas, which store large amounts of carbon and experience pronounced climatic warming. Despite the long-term standing paradigm assumes that cold ecosystems primarily have nitrogen deficiency, large-scale empirical tests of microbial nutrient limitation are lacking. Here we assessed the potential microbial nutrient limitation across the Tibetan alpine permafrost region, using the combination of enzymatic and elemental stoichiometry, genes abundance and fertilization method. In contrast with the traditional view, the four independent approaches congruently detected widespread microbial nitrogen and phosphorus co-limitation in both the surface soil and deep permafrost deposits, with stronger limitation in the topsoil. Further analysis revealed that soil resources stoichiometry and microbial community composition were the two best predictors of the magnitude of microbial nutrient limitation. High ratio of available soil carbon to nutrient and low fungal/bacterial ratio corresponded to strong microbial nutrient limitation. These findings suggest that warming-induced enhancement in soil nutrient availability could stimulate microbial activity, and probably amplify soil carbon losses from permafrost areas.     

岷江干旱河谷优势灌丛对土壤微生物群落组成的影响

通过调查岷江干旱河谷两河口、飞虹、撮箕和牟托4个样地优势灌丛及其灌丛间空地的表土土壤物理化学性质和微生物群落组成,探讨植物灌丛群落对土壤微生物群落组成的影响。研究发现不同灌丛种类对土壤微生物群落组成以及土壤物理化学性质并没有显著影响,而同一样地灌丛与空地间的差异却较为显著。灌丛下比空地土壤中具有更高的有机质、养分含量,更高的土壤含水量和更低的容重,而灌丛下相对富集的养分资源是造成灌丛与空地间微生物群落组成差异的主要原因。不同样地影响微生物群落的主要因子存在一定差异,但与氮相关的因子(总氮、有效氮、碳/氮比)对土壤微生物群落着非常重要的影响,特别是对土壤微生物群落总生物量和细菌类群(革兰氏阳性菌、革兰氏阴性菌、细菌等)。虽然不同灌丛和空地下土壤中细菌群落都没有显著地变化,但真菌和菌根真菌却明显的在灌丛下富集。在飞虹和牟托样地,总磷和碳/磷比与真菌类群,主要指真菌和菌根真菌,表现出显著正相关性,这或许反映了真菌类群对于该区域磷循环的重要作用。研究结果揭示了灌丛植被在干旱河谷地区地下生态系统中的重要作用,以及氮、磷这两种养分元素对土壤微生物群落的重要影响。同时,未来对于干旱河谷地区植物-土壤关系的研究应该关注真菌和菌根真菌类群的作用。     

Cascading influence of inorganic nitrogen sources on DOM production,composition, lability and microbial community structure in the open ocean

Nitrogen frequently limits oceanic photosynthesis and the availability of inorganic nitrogen sources in the surface oceans is shifting with global change. We evaluated the potential for abrupt increases in inorganic N sources to induce cascading effects on dissolved organic matter (DOM) and microbial communities in the surface ocean. We collected water from 5 m depth in the central North Pacific and amended duplicate 20 liter polycarbonate carboys with nitrate or ammonium, tracking planktonic carbon fixation, DOM production, DOM composition and microbial community structure responses over 1 week relative to controls. Both nitrogen sources stimulated bulk phytoplankton, bacterial and DOM production and enriched Synechococcus and Flavobacteriaceae; ammonium enriched for oligotrophic Actinobacteria OM1 and Gammaproteobacteria KI89A clades while nitrate enriched Gammaproteobacteria SAR86, SAR92 and OM60 clades. DOM resulting from both N enrichments was more labile and stimulated growth of copiotrophic Gammaproteobacteria (Alteromonadaceae and Oceanospirillaceae) and Alphaproteobacteria (Rhodobacteraceae and Hyphomonadaceae) in weeklong dark incubations relative to controls. Our study illustrates how nitrogen pulses may have direct and cascading effects on DOM composition and microbial community dynamics in the open ocean.     

Taxonomic and regional patterns in benthic macroinvertebrate elemental composition in streams

1. Ecological stoichiometry has been used to better understand dynamics in consumer growth and the role of consumer‐recycled nutrients because it focuses on more than one element. Most research has focused on pelagic rather than benthic consumers. Variation in elemental composition among benthic consumer taxa would suggest that taxa differ in their susceptibility to nutrient limitation or in their role in recycling nutrients. 2. We collected benthic macroinvertebrates from streams in two regions (Indiana–Michigan and Wisconsin, U.S.A.) to examine taxonomic and regional variation in benthic macroinvertebrate body carbon (C), nitrogen (N), and phosphorus (P) concentrations and ratios. 3. Elemental composition varied little within taxa common to both regions. In contrast, elemental composition differed greatly among taxa and appeared to be related to phylogeny. The elemental composition of macroinvertebrates clustered into three distinct groups: insects, mollusks, and crustaceans. To a lesser extent, insects and mollusks also differed in elemental composition among genera. 4. Functional feeding groups (FFGs) differed in elemental composition, with predators having a higher N content than other groups. Substantial elemental imbalances between C and N were found between most primary consumers and their likely food sources, and the magnitude of the imbalance depended in part on the FFG. 5. Our results support an assumption of most ecological stoichiometry models that, within a species, the elemental composition of aquatic invertebrates is relatively constant. Variation in elemental composition among taxa at various higher taxonomic levels suggests that susceptibility of stream invertebrates to nutrient limitation and their role in nutrient cycling will strongly depend on phylogeny.     

Microbial community responses to nitrogen- and phosphorus- deficient nutrient inputs: microplankton dynamics and biochemical characterization

Natural plankton communities from Masnou, a locality 20 km northof Barcelona (NW Mediterranean coast), were enclosed in 30 lmicrocosms to test the effect of different availability of nitrogen(N) and phosphorus (P) on the biomass of the main microplanktongroups, and the biochemical composition (DNA, protein and chlorophyllconcentration) of microbial communities. Immediately after enclosurein microcosms, three different nutrient enrichments were performed:N-deficient, P-deficient and nutrient-balanced. N and P deficienciesaffected the structure and the biochemical composition of themicrobial communities. Phytoplankton assemblages showed similartemporal patterns under the three nutrient treatments, althoughthe relative contribution of the different groups was notablyaffected. The lowest DNA concentration was measured in the P-deficienttreatment, suggesting that P availability imposes the limitson the DNA levels in the ecosystem. The availability of N inthe P-deficient microcosms allowed relatively high synthesisof chlorophyll and protein until the end of the experiment.Significantly high chlorophyll: DNA and protein: DNA ratioscharacterized the P-deficient treatment (where N was available)compared to the N-deficient microcosms. From the results obtained,we suggest that the protein: DNA ratios may constitute a biochemicalindicator of the P versus N availability in natural ecosystems.     

GeoChip 4: a functional gene‐array‐based high‐throughput environmental technology for microbial community analysis

Micro‐organisms play critical roles in many important biogeochemical processes in the Earth's biosphere. However, understanding and characterizing the functional capacity of microbial communities are still difficult due to the extremely diverse and often uncultivable nature of most micro‐organisms. In this study, we developed a new functional gene array, GeoChip 4, for analysing the functional diversity, composition, structure, metabolic potential/activity and dynamics of microbial communities. GeoChip 4 contained approximately 82 000 probes covering 141 995 coding sequences from 410 functional gene families related to microbial carbon (C), nitrogen (N), sulphur (S), and phosphorus (P) cycling, energy metabolism, antibiotic resistance, metal resistance/reduction, organic remediation, stress responses, bacteriophage and virulence. A total of 173 archaeal, 4138 bacterial, 404 eukaryotic and 252 viral strains were targeted, providing the ability to analyse targeted functional gene families of micro‐organisms included in all four domains. Experimental assessment using different amounts of DNA suggested that as little as 500 ng environmental DNA was required for good hybridization, and the signal intensities detected were well correlated with the DNA amount used. GeoChip 4 was then applied to study the effect of long‐term warming on soil microbial communities at a Central Oklahoma site, with results indicating that microbial communities respond to long‐term warming by enriching carbon degradation, nutrient cycling (nitrogen and phosphorous) and stress response gene families. To the best of our knowledge, GeoChip 4 is the most comprehensive functional gene array for microbial community analysis.     

Soil microorganisms respond to five years of climate change manipulations and elevated atmospheric CO2 in a temperate heath ecosystem

Background and aims

Soil microbial responses to global change can affect organic matter turnover and nutrient cycling thereby altering the overall ecosystem functioning. In a large-scale experiment, we investigated the impact of 5 years of climate change and elevated atmospheric CO₂ on soil microorganisms and nutrient availability in a temperate heathland.

Methods

The future climate was simulated by increased soil temperature (+0.3 °C), extended pre-summer drought (excluding 5–8 % of the annual precipitation) and elevated CO₂ (+130 ppm) in a factorial design. Soil organic matter and nutrient pools were analysed and linked to microbial measures by quantitative PCR of bacteria and fungi, chloroform fumigation extraction, and substrate-induced respiration to assess their impact of climate change on nutrient availability.

Results

Warming resulted in higher measures of fungi and bacteria, of microbial biomass and of microbial growth potential, however, this did not reduce the availability of nitrogen or phosphorus in the soil. Elevated CO₂ did not directly affect the microbial measures or nutrient pools, whereas drought shifted the microbial community towards a higher fungal dominance.

Conclusions

Although we were not able to show strong interactive effects of the global change factors, warming and drought changed both nutrient availability and microbial community composition in the heathland soil, which could alter the ecosystem carbon and nutrient flow in the long-term.     

VITCOMIC2: visualization tool for the phylogenetic composition of microbial communities based on 16S rRNA gene amplicons and metagenomic shotgun sequencing

Background

The 16S rRNA gene-based amplicon sequencing analysis is widely used to determine the taxonomic composition of microbial communities. Once the taxonomic composition of each community is obtained, evolutionary relationships among taxa are inferred by a phylogenetic tree. Thus, the combined representation of taxonomic composition and phylogenetic relationships among taxa is a powerful method for understanding microbial community structure; however, applying phylogenetic tree-based representation with information on the abundance of thousands or more taxa in each community is a difficult task. For this purpose, we previously developed the tool VITCOMIC (VIsualization tool for Taxonomic COmpositions of MIcrobial Community), which is based on the genome-sequenced microbes’ phylogenetic information. Here, we introduce VITCOMIC2, which incorporates substantive improvements over VITCOMIC that were necessary to address several issues associated with 16S rRNA gene-based analysis of microbial communities.

Results

We developed VITCOMIC2 to provide (i) sequence identity searches against broad reference taxa including uncultured taxa; (ii) normalization of 16S rRNA gene copy number differences among taxa; (iii) rapid sequence identity searches by applying the graphics processing unit-based sequence identity search tool CLAST; (iv) accurate taxonomic composition inference and nearly full-length 16S rRNA gene sequence reconstructions for metagenomic shotgun sequencing; and (v) an interactive user interface for simultaneous representation of the taxonomic composition of microbial communities and phylogenetic relationships among taxa. We validated the accuracy of processes (ii) and (iv) by using metagenomic shotgun sequencing data from a mock microbial community.

Conclusions

The improvements incorporated into VITCOMIC2 enable users to acquire an intuitive understanding of microbial community composition based on the 16S rRNA gene sequence data obtained from both metagenomic shotgun and amplicon sequencing.

     

Identification of General Patterns of Nutrient and Labile Carbon Control on Soil Carbon Dynamics Across a Successional Gradient

Carbon (C) inputs and nutrient availability are known to affect soil organic carbon (SOC) stocks. However, general rules regarding the operation of these factors across a range of soil nutrient availabilities and substrate qualities are unidentified. “Priming” (stimulated decomposition by labile C inputs) and ‘preferential substrate utilization’ (retarded decomposition due to shifts in community composition towards microbes that do not mineralize SOC) are two hypotheses to explain effects of labile C additions on SOC dynamics. For effects of nutrient additions (nitrogen and phosphorus) on SOC dynamics, the stoichiometric (faster decomposition of materials of low carbon-to-nutrient ratios) and ‘microbial mining’ (that is, reduced breakdown of recalcitrant C forms for nutrients under fertile conditions) hypotheses have been proposed. Using the natural gradient of soil nutrient availability and substrate quality of a chronosequence, combined with labile C and nutrient amendments, we explored the support for these contrasting hypotheses. Additions of labile C, nitrogen (N), phosphorus (P), and combinations of C and N and C and P were applied to three sites: 2-year fallow grassland, mature grassland and forest, and the effects of site and nutrient additions on litter decomposition and soil C dynamics were assessed. The response to C addition supported the preferential substrate hypothesis for easily degradable litter C and the priming hypothesis for SOC, but only in nitrogen-enriched soils of the forest site. Responses to N addition supported the microbial mining hypothesis irrespective of C substrate (litter or SOC), but only in the forest site. Further, P addition effects on SOC support the stoichiometric hypothesis; P availability appeared key to soil C release (priming) in the forest site if labile C and N is available. These results clearly link previously contrasting hypotheses of the factors controlling SOC with the natural gradient in litter quality and nutrient availability that exists in ecosystems at different successional stages. A holistic theory that incorporates this variability of responses, due to different mechanisms, depending on nutrient availability and substrate quality is essential for devising management strategies to safeguard soil C stocks.