Bacterial Diversity in the Rhizosphere of Cucumbers Grown in Soils Covering a Wide Range of Cucumber Cropping Histories and Environmental Conditions

Rhizosphere microorganisms in soils are important for plant growth. However, the importance of rhizosphere microorganisms is still underestimated since many microorganisms associated with plant roots cannot be cultured and since the microbial diversity in the rhizosphere can be influenced by several factors, such as the cropping history, biogeography, and agricultural practice. Here, we characterized the rhizosphere bacterial diversity of cucumber plants grown in soils covering a wide range of cucumber cropping histories and environmental conditions by using pyrosequencing of bacterial 16S rRNA genes. We also tested the effects of compost addition and/or bacterial inoculation on the bacterial diversity in the rhizosphere. We identified an average of approximately 8,883 reads per sample, corresponding to around 4,993 molecular operational taxonomic units per sample. The Proteobacteria was the most abundant phylum in almost all soils. The abundances of the phyla Bacteroidetes, Actinobacteria, Firmicutes, Acidobacteria, and Verrucomicrobia varied among the samples, and together with Proteobacteria, these phyla were the six most abundant phyla in almost all analyzed samples. Analyzing all the sample libraries together, the predominant genera found were Flavobacterium, Ohtaekwangia, Opitutus, Gp6, Steroidobacter, and Acidovorax. Overall, compost and microbial amendments increased shoot biomass when compared to untreated soils. However, compost addition decreased the bacterial α-diversity in most soils (but for three soils compost increased diversity), and no statistical effect of microbial amendment on the bacterial α-diversity was found. Moreover, soil amendments did not significantly influence the bacterial β-diversity. Soil organic content appeared more important than compost and microbial amendments in shaping the structure of bacterial communities in the rhizosphere of cucumber.     

Aerobic endospore-forming bacteria isolated from Antarctic soils as producers of bioactive compounds of industrial interest

Spore-forming bacteria are known to produce various enzymes and bioproducts valuable to different industries and to bear the harsh conditions found in the Antarctic environment. However, aerobic or facultative spore-forming bacterial communities found in maritime Antarctic soils yet remain poorly studied. In this study, 80 spore-forming and cold-adapted bacterial strains were isolated from nine different soil samples of King George Island, in maritime Antarctica, and further clustered into amplified ribosomal DNA restriction analysis groups within each soil. Representative strains were then identified as belonging to Bacillus, Rummeliibacillus, Paenibacillus and Sporosarcina by 16S rRNA gene sequencing. The ability to produce extracellular enzymes, antimicrobial substances and biosurfactants was determined in all isolates. The enzymatic activities most frequently found among the isolates were as follows: esterase (45 %), caseinase (30 %), amylase (16.2 %) and gelatinase (15 %). Biosurfactant production was detected in 25 % of the isolates. The growth inhibition of methicillin-resistant Staphylococcus aureus was observed in 13.7 % of the strains tested, but only two strains inhibited the growth of Candida albicans. The isolated spore-forming bacterial species were also compared with the characteristics of the different Antarctic soils sampled based on their physicochemical properties, showing that pH, C and P were the main factors correlated with the distribution of this group of bacteria in the Antarctic soils studied. These Antarctic endospore-forming bacterial strains may have a potential for industrial processes occurring at low temperatures.     

Identification of bacteria associated with underground parts of Crocus sativus by 16S rRNA gene targeted metagenomic approach

Saffron (Crocus sativus L), an autumn-flowering perennial sterile plant, reproduces vegetatively by underground corms. Saffron has biannual corm–root cycle that makes it an interesting candidate to study microbial dynamics in its rhizosphere and cormosphere (area under influence of corm). Culture independent 16S rRNA gene metagenomic study of rhizosphere and cormosphere of Saffron during flowering stage revealed presence of 22 genera but none of the genus was common in all the three samples. Bulk soil bacterial community was represented by 13 genera with Acidobacteria being dominant. In rhizosphere, out of eight different genera identified, Pseudomonas was the most dominant genus. Cormosphere bacteria comprised of six different genera, dominated by the genus Pantoea. This study revealed that the bacterial composition of all the three samples is significantly different (P < 0.05) from each other. This is the first report on the identification of bacteria associated with rhizosphere, cormosphere and bulk soil of Saffron, using cultivation independent 16S rRNA gene targeted metagenomic approach.     

Spatial distribution of bacterial communities in high-altitude freshwater wetland sediment

Sediment microorganisms play a crucial role in a variety of biogeochemical processes in freshwater ecosystems. The objective of the current study was to investigate the spatial distribution of sediment bacterial community structure in Luoshijiang Wetland, located in Yunnan–Kweichow Plateau (China). Wetland sediments at different sites and depths were collected. Clone library analysis indicates bacterial communities varied with both sampling site and sediment depth. A total of fourteen bacterial phyla were identified in sediment samples, including Proteobacteria, Acidobacteria, Actinobacteria, Armatimonadetes, Bacteroidetes, Chlorobi, Chloroflexi, Cyanobacteria, Firmicutes, Gemmatimonadetes, Nitrospirae, Planctomycetes, Spirochaetes, and Verrucomicrobia. Proteobacteria (mainly Betaproteobacteria and Deltaproteobacteria) predominated in wetland sediments. Moreover, the proportions of Alphaproteobacteria, Acidobacteria, Bacteroidetes, Gemmatimonadete, and Planctomycetes were significantly correlated with chemical properties.     

Environmental Heterogeneity and Microbial Inheritance Influence Sponge-Associated Bacterial Composition of Spongia lamella

Sponges are important components of marine benthic communities. High microbial abundance sponges host a large diversity of associated microbial assemblages. However, the dynamics of such assemblages are still poorly known. In this study, we investigated whether bacterial assemblages present in Spongia lamella remained constant or changed as a function of the environment and life cycle. Sponges were collected in multiple locations and at different times of the year in the western Mediterranean Sea and in nearby Atlantic Ocean to cover heterogeneous environmental variability. Co-occurring adult sponges and offsprings were compared at two of the sites. To explore the composition and abundance of the main bacteria present in the sponge mesohyl, embryos, and larvae, we applied both 16S rRNA gene-denaturing gradient gel electrophoresis (DGGE) and sequencing of excised DGGE bands and quantitative polymerase chain reactions (qPCR). On average, the overall core bacterial assemblage showed over 60 % similarity. The associated bacterial assemblage fingerprints varied both within and between sponge populations, and the abundance of specific bacterial taxa assessed by qPCR significantly differed among sponge populations and between adult sponge and offsprings (higher proportions of Actinobacteria in the latter). Sequences showed between 92 and 100 % identity to sequences previously reported in GenBank, and all were affiliated with uncultured invertebrate bacterial symbionts (mainly sponges). Sequences were mainly related to Chloroflexi and Acidobacteria and a few to Actinobacteria and Bacteroidetes. Additional populations may have been present under detection limits. Overall, these results support that both ecological and biological sponge features may shape the composition of endobiont bacterial communities in S. lamella.     

Progressive and retrogressive ecosystem development coincide with soil bacterial community change in a dune system under lowland temperate rainforest in New Zealand

Background

It is established that plant communities show patterns of change linked to progressive and retrogressive stages of ecosystem development. It is not known, however, whether bacterial communities also show similar patterns of change associated with long-term ecosystem development.

Methods

We studied soil bacterial communities along a 6,500 year dune chronosequence under lowland temperate rain forest at Haast, New Zealand. Pyrosequencing of 16S rRNA genes was used to observe structural change in bacterial communities during the process of pedogenesis and ecosystem development.

Results

Bacterial communities showed patterns of change during pedogenesis, with the largest change during the first several hundred years after dune stabilization. The most abundant bacterial taxa were Alphaproteobacteria, Actinobacteria and Acidobacteria. These include taxa most closely related to nitrogen-fixing bacteria, and suggest heterotrophic nitrogen input may be important throughout the chronosequence. Changes in bacterial community structure were related to changes in several soil properties, including total phosphorus, C:N ratio, and pH. The Bacteroidetes, Actinobacteria, Cyanobacteria, Firmicutes, and Betaproteobacteria all showed a general decline in abundance as pedogenesis proceeded, while Acidobacteria, Alphaproteobacteria, and Plantctomycetes tended to increase as soils aged.

Conclusions

There were trends in the dynamics of bacterial community composition and structure in soil during ecosystem development. Bacterial communities changed in ways that appear to be consistent with a model of ecosystem progression and retrogression, perhaps indicating fundamental processes underpin patterns of below and above-ground community change during ecosystem development.     

Effects of variation in precipitation on the distribution of soil bacterial diversity in the primitive Korean pine and broadleaved forests

Patterns of precipitation have changed as a result of climate change and will potentially keep changing in the future. Therefore, it is critical to understand how ecosystem processes will respond to the variation of precipitation. However, compared to aboveground processes, the effects of precipitation change on soil microorganisms remain poorly understood. Changbai Mountain is an ideal area to study the responses of temperate forests to the variations in precipitation. In this study, we conducted a manipulation experiment to simulation variation of precipitation in the virgin, broad-leaved Korean pine mixed forest in Changbai Mountain. Plots were designed to increase precipitation by 30 % [increased (+)] or decrease precipitation by 30 % [decreased (?)]. We analyzed differences in the diversity of the bacterial community in surface bulk soils (0–5 and 5–10 cm) and rhizosphere soils between precipitation treatments, including control. Bacteria were identified using the high-throughput 454 sequencing method. We obtained a total 271,496 optimized sequences, with a mean value of 33,242 (±1,412.39) sequences for each soil sample. Being the same among the sample plots with different precipitation levels, the dominant bacterial communities were Proteobacteria, Acidobacteria, Actinobacteria, Planctomycetes, and Chloroflexi. Bacterial diversity and abundance declined with increasing soil depth. In the bulk soil of 0–5 cm, the bacterial diversity and abundance was the highest in the control plots and the lowest in plots with reduced precipitation. However, in the soil of 5–10 cm, the diversity and abundance of bacteria was the highest in the plots of increased precipitation and the lowest in the control plots. Bacterial diversity and abundance in rhizosphere soils decreased with increased precipitation. This result implies that variation in precipitation did not change the composition of the dominant bacterial communities but affected bacterial abundance and the response patterns of the dominant communities to variation in precipitation.     

Biodiversity and physiological characteristics of Antarctic and Arctic lichens-associated bacteria

The diversity and physiological characteristics of culturable bacteria associated with lichens from different habitats of the Arctic and Antarctica were investigated. The 68 retrieved isolates could be grouped on the basis of their 16S rRNA gene sequences into 26 phylotypes affiliated with the phyla Actinobacteria, Bacteroidetes, Deinococcus-Thermus, and Firmicutes and with the classes Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria. Isolates belonging to the Alphaproteobacteria were the most abundant, followed by those belonging to Actinobacteria, Betaproteobacteria, Gammaproteobacteria, Bacteroidetes, Firmicutes, and Deinococcus-Thermus. Phylogenetic analysis showed that approximately 21 % of the total isolates represented a potentially novel species or genus (≤97 % sequence similarity). Strains belonging to the genera Sphingomonas, Frondihabitans, Hymenobacter, and Burkholderia were recovered from lichen samples from both geographic locations, implying common and important bacterial functions within lichens. Extracellular protease activities were detected in six isolates, affiliated with Burkholderia, Frondihabitans, Hymenobacter, Pseudomonas, and Rhodanobacter. Extracellular lipase activities were detected in 37 isolates of the genera Burkholderia, Deinococcus, Frondihabitans, Pseudomonas, Rhodanobacter, Sphingomonas, and Subtercola. This is the first report on the culturable bacterial diversity present within lichens from Arctic and Antarctica and the isolates described herein are valuable resources to decode the functional and ecological roles of bacteria within lichens. In addition, the low similarity (≤97 %) of the recovered isolates to known species and their production of cold-active enzymes together suggest that lichens are noteworthy sources of novel bacterial strains for use in biotechnological applications.     

Bacterial community analysis in chlorpyrifos enrichment cultures via DGGE and use of bacterial consortium for CP biodegradation

The organophosphate pesticide chlorpyrifos (CP) has been used extensively since the 1960s for insect control. However, its toxic effects on mammals and persistence in environment necessitate its removal from contaminated sites, biodegradation studies of CP-degrading microbes are therefore of immense importance. Samples from a Pakistani agricultural soil with an extensive history of CP application were used to prepare enrichment cultures using CP as sole carbon source for bacterial community analysis and isolation of CP metabolizing bacteria. Bacterial community analysis (denaturing gradient gel electrophoresis) revealed that the dominant genera enriched under these conditions were Pseudomonas, Acinetobacter and Stenotrophomonas, along with lower numbers of Sphingomonas, Agrobacterium and Burkholderia. Furthermore, it revealed that members of Bacteroidetes, Firmicutes, α- and γ-Proteobacteria and Actinobacteria were present at initial steps of enrichment whereas β-Proteobacteria appeared in later steps and only Proteobacteria were selected by enrichment culturing. However, when CP-degrading strains were isolated from this enrichment culture, the most active organisms were strains of Acinetobacter calcoaceticus, Pseudomonas mendocina and Pseudomonas aeruginosa. These strains degraded 6–7.4 mg L^?1 day^?1 of CP when cultivated in mineral medium, while the consortium of all four strains degraded 9.2 mg L^?1 day^?1 of CP (100 mg L^?1). Addition of glucose as an additional C source increased the degradation capacity by 8–14 %. After inoculation of contaminated soil with CP (200 mg kg^?1) disappearance rates were 3.83–4.30 mg kg^?1 day^?1 for individual strains and 4.76 mg kg^?1 day^?1 for the consortium. These results indicate that these organisms are involved in the degradation of CP in soil and represent valuable candidates for in situ bioremediation of contaminated soils and waters.     

Bacterial communities associated with the pitcher fluids of three Nepenthes (Nepenthaceae) pitcher plant species growing in the wild

Nepenthes pitcher plants produce modified jug-shaped leaves to attract, trap and digest insect prey. We used 16S rDNA cloning and sequencing to compare bacterial communities in pitcher fluids of each of three species, namely Nepenthes ampullaria, Nepenthes gracilis and Nepenthes mirabilis, growing in the wild. In contrast to previous greenhouse-based studies, we found that both opened and unopened pitchers harbored bacterial DNA. Pitchers of N. mirabilis had higher bacterial diversity as compared to other Nepenthes species. The composition of the bacterial communities could be different between pitcher types for N. mirabilis (ANOSIM: R = 0.340, p < 0.05). Other Nepenthes species had similar bacterial composition between pitcher types. SIMPER showed that more than 50 % of the bacterial taxa identified from the open pitchers of N. mirabilis were not found in other groups. Our study suggests that bacteria in N. mirabilis are divided into native and nonnative groups.     

Niche partitioning of bacterial communities in biological crusts and soils under grasses,shrubs and trees in the Kalahari

The Kalahari of southern Africa is characterised by sparse vegetation interspersed with microbe-dominated biological soil crusts (BSC) which deliver a range of ecosystem services including soil stabilisation and carbon fixation. We characterised the bacterial communities of BSCs (0–1 cm depth) and the subsurface soil (1–2 cm depth) in an area typical of lightly grazed Kalahari rangelands, composed of grasses, shrubs, and trees. Our data add substantially to the limited amount of existing knowledge concerning BSC microbial community structure, by providing the first bacterial community analyses of both BSCs and subsurface soils of the Kalahari region based on a high throughput 16S ribosomal RNA gene sequencing approach. BSC bacterial communities were distinct with respect to vegetation type and soil depth, and varied in relation to soil carbon, nitrogen, and surface temperature. Cyanobacteria were predominant in the grass interspaces at the soil surface (0–1 cm) but rare in subsurface soils (1–2 cm depth) and under the shrubs and trees. Bacteroidetes were significantly more abundant in surface soils of all areas even in the absence of a consolidated crust, whilst subsurface soils yielded more sequences affiliated to Acidobacteria, Actinobacteria, Chloroflexi, and Firmicutes. The common detection of vertical stratification, even in disturbed sites, suggests a strong potential for BSC recovery after physical disruption, however severe depletion of Cyanobacteria near trees and shrubs may limit the potential for natural BSC regeneration in heavily shrub-encroached areas.     

Effects of polycyclic aromatic hydrocarbons on microbial community structure and PAH ring hydroxylating dioxygenase gene abundance in soil

Development of successful bioremediation strategies for environments contaminated with recalcitrant pollutants requires in-depth knowledge of the microorganisms and microbial processes involved in degradation. The response of soil microbial communities to three polycyclic aromatic hydrocarbons, phenanthrene (3-ring), fluoranthene (4-ring) and benzo(a)pyrene (5-ring), was examined. Profiles of bacterial, archaeal and fungal communities were generated using molecular fingerprinting techniques (TRFLP, ARISA) and multivariate statistical tools were employed to interpret the effect of PAHs on community dynamics and composition. The extent and rate of PAH removal was directly related to the chemical structure, with the 5-ring PAH benzo(a)pyrene degraded more slowly than phenathrene or fluoranthene. Bacterial, archaeal and fungal communities were all significantly affected by PAH amendment, time and their interaction. Based on analysis of clone libraries, Actinobacteria appeared to dominate in fluoranthene amended soil, although they also represented a significant portion of the diversity in phenanthrene amended and unamended soils. In addition there appeared to be more γ-Proteobacteria and less Bacteroidetes in soil amended with either PAH compared to the control. The soil bacterial community clearly possessed the potential to degrade PAHs as evidenced by the abundance of PAH ring hydroxylating (PAH-RHDα) genes from both gram negative (GN) and gram positive (GP) bacteria in PAH-amended and control soils. Although the dioxygenase gene from GP bacteria was less abundant in soil than the gene associated with GN bacteria, significant (p < 0.001) increases in the abundance of the GP PAH-RHDα gene were observed during phenanthrene and fluoranthene degradation, whereas there was no significant difference in the abundance of the GN PAH-RHDα gene during the course of the experiment. Few studies to-date have examined the effect of pollutants on more than one microbial community in soil. The current study provides information on the response of soil bacterial, archaeal and fungal communities during the degradation of three priority pollutants and contributes to a knowledge base that can inform the development of effective bioremediation strategies for contaminated sites.     

Particulate matter and bacteria characteristics of the Middle East Dust (MED) storms over Ahvaz,Iran

The Middle East Dust storms have greatly affected the south and west parts of Iran during the last decade. The main purpose of this study was to examine and compare culturable airborne bacteria concentration in particulate matter (PM) during normal, semi-dust, and dust event days in different places and seasons in Ahvaz from November 2011 to May 2012. Sampling was performed every 6 days and on dust event days at different sampling stations. The overall mean concentrations of PM₁₀, PM_2.5, and PM₁ for the entire study period were 598.92, 114.8, and 34.5 μg/m³, respectively. The PM concentrations during the dust event days were much higher than normal and semi-dust event days. The highest mean PM concentrations were observed in March 2011. The low PM_2.5/PM₁₀ ratios indicate that these PM are mostly originating from natural sources such as dust storms. The overall mean concentration of total bacteria during the study period was 620.6 CFU/m³. The greatest bacterial concentrations were observed during dust event days and at areas with high traffic and more human activities compared with normal days and greener areas. The percentage of gram-positive bacteria was significantly higher than that during the study period (89 vs 11 %). During this study, 26 genera of culturable bacteria were identified from all the sampling stations. The most dominant genera in all sampling stations were Streptomyces, Bacillus, Kocuria, Corynebacterium, and Paenibacillus. The results also showed that there were positive correlations between PM and bacterial concentrations during the study period (p < 0.05).     

Effects of inorganic nanoparticles on viability and catabolic activities of Agrobacterium sp. PH-08 during biodegradation of dibenzofuran

This study investigated the cytotoxicity, genotoxicity, and growth inhibition effects of four different inorganic nanoparticles (NPs) such as aluminum (nAl), iron (nFe), nickel (nNi), and zinc (nZn) on a dibenzofuran (DF) degrading bacterium Agrobacterium sp. PH-08. NP (0–1,000 mg L^?1) -treated bacterial cells were assessed for cytotoxicity, genotoxicity, growth and biodegradation activities at biochemical and molecular levels. In an aqueous system, the bacterial cells treated with nAl, nZn and nNi at 500 mg L^?1 showed significant reduction in cell viability (30–93.6 %, p < 0.05), while nFe had no significant inhibition on bacterial cell viability. In the presence of nAl, nZn and nNi, the cells exhibited elevated levels of reactive oxygen species (ROS), DNA damage and cell death. Furthermore, NP exposure showed significant (p < 0.05) impairment in DF and catechol biodegradation activities. The reduction in DF biodegradation was ranged about 71.7–91.6 % with single NPs treatments while reached up to 96.3 % with a mixture of NPs. Molecular and biochemical investigations also clearly revealed that NP exposure drastically affected the catechol-2,3-dioxygenase activities and its gene (c23o) expression. However, no significant inhibition was observed in nFe treatment. The bacterial extracellular polymeric materials and by-products from DF degradation can be assumed as key factors in diminishing the toxic effects of NPs, especially for nFe. This study clearly demonstrates the impact of single and mixed NPs on the microbial catabolism of xenobiotic-degrading bacteria at biochemical and molecular levels. This is the first study on estimating the impact of mixed NPs on microbial biodegradation.     

Kinetics of nitrification in a fixed biofilm reactor using dewatered sludge-fly ash composite ceramic particle as a supporting medium

A mathematical model system was derived to describe the kinetics of ammonium nitrification in a fixed biofilm reactor using dewatered sludge-fly ash composite ceramic particle as a supporting medium. The model incorporates diffusive mass transport and Monod kinetics. The model was solved using a combination of the orthogonal collocation method and Gear’s method. A batch test was conducted to observe the nitrification of ammonium-nitrogen ( \({\text{NH}}_{4}^{ + }\) -N) and the growth of nitrifying biomass. The compositions of nitrifying bacterial community in the batch kinetic test were analyzed using PCR–DGGE method. The experimental results show that the most staining intensity abundance of bands occurred on day 2.75 with the highest biomass concentration of 46.5 mg/L. Chemostat kinetic tests were performed independently to evaluate the biokinetic parameters used in the model prediction. In the column test, the removal efficiency of \({\text{NH}}_{4}^{ + }\) -N was approximately 96 % while the concentration of suspended nitrifying biomass was approximately 16 mg VSS/L and model-predicted biofilm thickness reached up to 0.21 cm in the steady state. The profiles of denaturing gradient gel electrophoresis (DGGE) of different microbial communities demonstrated that indigenous nitrifying bacteria (Nitrospira and Nitrobacter) existed and were the dominant species in the fixed biofilm process.     

The effect of soil water status on fertiliser, topsoil and subsoil phosphorus utilisation by wheat

Background and aims

Crop phosphorus (P) content is controlled by P uptake from both banded P fertiliser and from P throughout the soil profile. These P supply factors are in turn controlled by soil, climatic and plant factors. The aim of this experiment was to measure the contribution of fertiliser, topsoil and subsoil P to wheat plants under wet and dry growing season conditions.

Methods

An isotopic tracer technique was used to measure P uptake from fertiliser at seven agricultural field sites under wet and dry growing season conditions. At three of these sites a dual isotopic technique was used to distinguish between wheat uptake of P from fertiliser, topsoil (0–15 cm) and subsoil (below 15 cm).

Results

The amount of P fertiliser used by wheat was in the order of 3–30% of the P applied and increased with increasing rainfall. Topsoil P was the most important P source, but when sufficient P was present in the subsoil, P fertiliser addition stimulated the use of subsoil P.

Conclusions

Most crop P uptake was from the topsoil, however P fertiliser banded below the seed increased plant P uptake and stimulated the use of subsoil P in one soil type in a decile 7 (above average rainfall) growing season.     

Flavobacterium ahnfeltiae sp. nov., a new marine polysaccharide-degrading bacterium isolated from a Pacific red alga

A Gram-negative, aerobic, rod-shaped, motile by gliding and yellow-pigmented bacterium, designated strain 10Alg 130^T, that displayed the ability to destroy polysaccharides of red and brown algae, was isolated from the red alga Ahnfeltia tobuchiensis. The phylogenetic analysis based on 16S rRNA gene sequence placed the novel strain within the genus Flavobacterium, the type genus of the family Flavobacteriaceae, the phylum Bacteroidetes, with sequence similarities of 96.2 and 95.7 % to Flavobacterium jumunjiense KCTC 23618^T and Flavobacterium ponti CCUG 58402^T, and 95.3–92.5 % to other recognized Flavobacterium species. The prevalent fatty acids of strain 10Alg 130^T were iso-C_15:0, iso-C_15:0 3-OH, iso-C_17:0 3-OH, C_15:0 and iso-C_17:1ω9c. The polar lipid profile consisted of phosphatidylethanolamine, two unknown aminolipids and three unknown lipids. The DNA G+C content of the type strain was 34.3 mol%. The new isolate and the type strains of recognized species of the genus Flavobacterium could strongly be distinguished by a number of phenotypic characteristics. A combination of the genotypic and phenotypic data showed that the algal isolate represents a novel species of the genus Flavobacterium, for which the name Flavobacterium ahnfeltiae sp. nov. is proposed. The type strain is 10Alg 130^T (=KCTC 32467^T = KMM 6686^T).     

Comparative analysis of bacterial community-metagenomics in coastal Gulf of Mexico sediment microcosms following exposure to Macondo oil (MC252)

The indigenous bacterial communities in sediment microcosms from Dauphin Island (DI), Petit Bois Island (PB) and Perdido Pass (PP) of the coastal Gulf of Mexico were compared following treatment with Macondo oil (MC252) using pyrosequencing and culture-based approaches. After quality-based trimming, 28,991 partial 16S rRNA sequence reads were analyzed by rarefaction, confirming that analyses of bacterial communities were saturated with respect to species diversity. Changes in the relative abundances of Proteobacteria, Bacteroidetes and Firmicutes played an important role in structuring bacterial communities in oil-treated sediments. Proteobacteria were dominant in oil-treated samples, whereas Firmicutes and Bacteroidetes were either the second or the third most abundant taxa. Tenericutes, members of which are known for oil biodegradation, were detected shortly after treatment, and continued to increase in DI and PP sediments. Multivariate statistical analyses (ADONIS) revealed significant dissimilarity of bacterial communities between oil-treated and untreated samples and among locations. In addition, a similarity percentage analysis showed the contribution of each species to the contrast between untreated and oil-treated samples. PCR amplification using DNA from pure cultures of Exiguobacterium,  Pseudoalteromonas,  Halomonas and Dyadobacter, isolated from oil-treated microcosm sediments, produced amplicons similar to polycyclic aromatic hydrocarbon-degrading genes. In the context of the 2010 Macondo blowout, the results from our study demonstrated that the indigenous bacterial communities in coastal Gulf of Mexico sediment microcosms responded to the MC252 oil with altered community structure and species composition. The rapid proliferation of hydrocarbonoclastic bacteria suggests their involvement in the degradation of the spilt oil in the Gulf of Mexico ecosystem.     

Horizon-Specific Bacterial Community Composition of German Grassland Soils,as Revealed by Pyrosequencing-Based Analysis of 16S rRNA Genes

The diversity of bacteria in soil is enormous, and soil bacterial communities can vary greatly in structure. Here, we employed a pyrosequencing-based analysis of the V2-V3 16S rRNA gene region to characterize the overall and horizon-specific (A and B horizons) bacterial community compositions in nine grassland soils, which covered three different land use types. The entire data set comprised 752,838 sequences, 600,544 of which could be classified below the domain level. The average number of sequences per horizon was 41,824. The dominant taxonomic groups present in all samples and horizons were the Acidobacteria, Betaproteobacteria, Actinobacteria, Gammaproteobacteria, Alphaproteobacteria, Deltaproteobacteria, Chloroflexi, Firmicutes, and Bacteroidetes. Despite these overarching dominant taxa, the abundance, diversity, and composition of bacterial communities were horizon specific. In almost all cases, the estimated bacterial diversity (H′) was higher in the A horizons than in the corresponding B horizons. In addition, the H′ was positively correlated with the organic carbon content, the total nitrogen content, and the C-to-N ratio, which decreased with soil depth. It appeared that lower land use intensity results in higher bacterial diversity. The majority of sequences affiliated with the Actinobacteria, Bacteroidetes, Cyanobacteria, Fibrobacteres, Firmicutes, Spirochaetes, Verrucomicrobia, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria were derived from A horizons, whereas the majority of the sequences related to Acidobacteria, Chloroflexi, Gemmatimonadetes, Nitrospira, TM7, and WS3 originated from B horizons. The distribution of some bacterial phylogenetic groups and subgroups in the different horizons correlated with soil properties such as organic carbon content, total nitrogen content, or microbial biomass.Soil is probably the most complex microbial environment on Earth with respect to species richness and community size. The microbial richness in soils exceeds that of other environments (44) and is higher by orders of magnitude than the biodiversity of plants and animals. Cultivated soil or grassland soil contains an estimated 2 × 10⁹ prokaryotic cells per gram (12). Soil microbial communities are an important factor of agriculturally managed systems, as they are responsible for most nutrient transformations in soil and influence the above-ground plant diversity and productivity (53).To analyze the bacterial community in soils, most approaches target the 16S rRNA gene by PCR amplification and subsequent analysis employing sequencing of clone libraries (10, 24), denaturing gradient gel electrophoresis (DGGE) (38), or terminal restriction fragment length polymorphism (T-RFLP) (17, 52). Most of these approaches provided limited insights into the structure of soil bacterial communities, as the survey sizes and the number of compared sampling sites were small with respect to the enormous bacterial diversity present in different soil samples. For example, the reported clone libraries vary considerably in size, but small sample sizes (500 or fewer 16S rRNA gene sequences) are usually analyzed and employed for the theoretical estimation of species richness (39). This provides snapshots of the predominant bacterial community members, but phylogenetic groups that are present in a low abundance and which may possess important ecosystem functions are not assessed (47). In addition, it has been shown that rich sampling (several thousands of clones) of complex bacterial communities is required to perform robust measurements and estimations of community diversity parameters (37). Thus, the detection bias accompanying analyses of small sample sizes can lead to invalidated assumptions. Genetic profiling techniques such as DGGE and T-RFLP have high-throughput capability. These approaches allow researchers to unravel differences in community structure but are limited for assessing diversity (23, 40). To deeply survey the diversity and the composition of the bacterial communities within different soil samples, large-scale pyrosequencing of partial 16S rRNA genes has been employed recently. Previous pyrosequencing-based studies of soil (1, 30, 34, 43) have generated large data sets, which comprised 39,707 (30) to 152,359 (34) 16S rRNA partial gene sequences. Those studies provided comprehensive insights into the biogeography of bacterial soil communities and taxa that were present in a low abundance. However, all those studies focused on the analysis of microbial communities present in topsoil. The subsoil is also known to harbor an important part of the soil microbial biomass (18). It has been shown that the microbial population in the shallow subsurface is impacted by agricultural production to a similar extent as that in topsoil (5).In this study, we performed large-scale pyrosequencing-based analyses of 16S rRNA genes to assess the bacterial community composition in topsoil and the corresponding subsoil of nine different grassland sites in the Hainich region (Thuringia, Germany). To provide a high level of coverage at the species level (97% genetic distance) and minimize detection bias, we exceeded the above-described numbers of analyzed 16S rRNA gene sequences (752,838 in this study). To examine the impact of land use on bacterial diversity and community composition, the selected grassland sites covered a range of three different land use types, including samples from unfertilized pastures grazed by cattle, fertilized mown pastures grazed by cattle, and fertilized meadows. In many recent studies, surveys were focused on comprehensive analyses of a single soil or a few soil samples (1, 14, 37, 43). This allowed the determination of overall bacterial species richness and community composition, but the assessment of spatial patterns and environmental factors that drive these patterns is hampered by the limited number of examined soils. To assess spatial distribution and the impact of soil edaphic factors and land use on community structure, we used triplicate samples of each land use type from different locations. In addition, composite samples derived from five soil cores after the separation of soil horizons were employed.