Genetic Structure Of Bacterial Communities Research Articles

Relative Weight of Organic Waste Origin on Compost and Digestate 16S rRNA Gene Bacterial Profilings and Related Functional Inferences.

Even though organic waste (OW) recycling via anaerobic digestion (AD) and composting are increasingly used, little is known about the impact of OW origin (fecal matters and food and vegetable wastes) on the end products’ bacterial contents. The hypothesis of a predictable bacterial community structure in the end products according to the OW origin was tested. Nine OW treatment plants were selected to assess the genetic structure of bacterial communities found in raw OW according to their content in agricultural and urban wastes and to estimate their modifications through AD and composting. Two main bacterial community structures among raw OWs were observed and matched a differentiation according to the occurrences of urban chemical pollutants. Composting led to similar 16S rRNA gene OTU profiles whatever the OW origin. With a significant shift of about 140 genera (representing 50% of the bacteria), composting was confirmed to largely shape bacterial communities toward similar structures. The enriched taxa were found to be involved in detoxification and bioremediation activities. This process was found to be highly selective and favorable for bacterial specialists. Digestates showed that OTU profiles differentiated into two groups according to their relative content in agricultural (manure) and urban wastes (mainly activated sludge). About one third of the bacterial taxa was significantly affected by AD. In digestates of urban OW, this sorting led to an enrichment of 32 out of the 50 impacted genera, while for those produced from agricultural or mixed urban/agricultural OW (called central OW), a decay of 54 genera over 60 was observed. Bacteria from activated sludge appeared more fit for AD than those of other origins. Functional inferences showed AD enriched genera from all origins to share similar functional traits, e.g., chemoheterotrophy and fermentation, while being often taxonomically distinct. The main functional traits among the dominant genera in activated sludge supported a role in AD. Raw OW content in activated sludge was found to be a critical factor for predicting digestate bacterial contents. Composting generated highly predictable and specialized community patterns whatever the OW origin. AD and composting bacterial changes were driven by functional traits selected by physicochemical factors such as temperature and chemical pollutants.

Intrinsic factors of Peltigera lichens influence the structure of the associated soil bacterial microbiota.

Definition of lichens has evolved from bi(tri)partite associations to multi-species symbioses, where bacteria would play essential roles. Besides, although soil bacterial communities are known to be affected by edaphic factors, when lichens grow upon them these could become less preponderant. We hypothesized that the structure of both the lichen microbiota and the microbiota in the soil underneath lichens is shaped by lichen intrinsic and extrinsic factors. In this work, intrinsic factors corresponded to mycobiont and cyanobiont identities of Peltigera lichens, metabolite diversity and phenoloxidase activity and extrinsic factors involved the site of the forest where lichens grow. Likewise, the genetic and metabolic structure of the lichen and soil bacterial communities were analyzed by fingerprinting. Among the results, metabolite diversity was inversely related to the genetic structure of bacterial communities of lichens and soils, highlighting the far-reaching effect of these substances; while phenoloxidase activity was inversely related to the metabolic structure only of the lichen bacterial microbiota, presuming a more limited effect of the products of these enzymes. Soil bacterial microbiota was different depending on the site and, strikingly, according to the cyanobiont present in the lichen over them, which could indicate an influence of the photobiont metabolism on the availability of soil nutrients.

Water-extractable organic matter linked to soil physico-chemistry and microbiology at the regional scale

A better understanding of the links between dissolved organic matter and biogeochemical processes in soil could help in evaluating global soil dynamics. To assess the effects of land cover and parental material on soil biogeochemistry, we studied 120 soil samples collected from various ecosystems in Burgundy, France. The potential solubility and aromaticity of dissolved organic matter was characterised by pressurised hot-water extraction of organic carbon (PH-WEOC). Soil physico-chemical characteristics (pH, texture, soil carbon and nitrogen) were measured, as was the δ13C signature both in soils and in PH-WEOC. We also determined bacterial and fungal abundance and the genetic structure of bacterial communities. Our results show that the potential solubility of soil organic carbon is correlated to carbon and clay content in the soil. The aromaticity of PH-WEOC and its δ13C signature reflect differences in the decomposition pathways of soil organic matter and in the production of water-extractable organic compounds, in relation to land cover. The genetic structure of bacterial communities is related to soil texture and pH, and to PH-WEOC, revealing that water-extractable organic matter is closely related to the dynamics of bacterial communities. This comprehensive study, at the regional scale, thus provides better definition of the relationships between water-extractable organic matter and soil biogeochemical properties.

Multivariate analysis of soils: microbial biomass, metabolic activity, and bacterial‐community structure and their relationships with soil depth and type

AbstractA multivariate statistical approach based on a large data set of abiotic and biotic variables was used to classify four contrasting‐land‐use soils. Soil samples were collected at increasing depth from a calcareous agricultural soil, a temperate upland grassland soil, a moderately acidic agricultural soil, and an acidic pine forest soil. Analytical investigations were carried out by using a combination of conventional physical, chemical, and biochemical methods coupled with denaturing gradient gel electrophoresis (DGGE) community fingerprinting of PCR‐amplified 16S rRNA gene‐coding fragments from soil‐extracted total‐community DNA. The data set of soil physical, chemical, and biochemical variables was reduced in dimensionality by means of a principal‐component‐analysis (PCA) procedure. Compositional shifts in soil bacterial‐community structure were analyzed through a clustering algorithm that allowed identifying six main bacterial‐community clusters. DGGE fingerprinting clusters were further analyzed by discriminant analysis (DA) using extracted PCA components as explanatory variables. Soil organic matter–related pools (TOC, TN) and functionally related active pools (microbial biomass C and N, K2SO4‐extractable C) significantly decreased with soil depth, and resulted statistically linked to one other and positively related to enzymatic activities (acid phosphatase, arylsulfatase, β‐glucosidase, dehydrogenase, hydrolysis of fluorescein diacetate) and silt content. Besides organic‐C gradients, pedogenetic‐driven physico‐chemical properties, and possibly soil thermal and moisture regimes seemed to play a key role in regulating size and energetic ecophysiological status of soil microbial communities. DGGE analysis showed that contrasting horizons were conducive to the dominance of particular bacterial ribotypes. DA revealed that the bacterial‐community structure was mainly influenced by organic matter–related variables (TOC, TN, CEC, Cflush, Nflush, Extr‐C), chemical properties such as pH, CaCO3, and EC, together with textural properties. Results indicate that, beyond land use or plant cover, pedogenetic‐driven physico‐chemical conditions changing with soil type and depth are the key factors regulating microbial size and activity, and determining the genetic structure of bacterial community.

Intraspecific genotypic richness and relatedness predict the invasibility of microbial communities

Biological invasions can lead to extinction events in resident communities and compromise ecosystem functioning. We tested the effect of two widespread biodiversity measurements, genotypic richness and genotypic dissimilarity on community invasibility. We manipulated the genetic structure of bacterial communities (Pseudomonas fluorescens) and submitted them to invasion by Serratia liquefaciens. We show that the two diversity measures impact on invasibility via distinct and additive mechanisms. Genotypic dissimilarity of the resident communities linearly increased productivity and in parallel decreased invasion success, indicating that high dissimilarity prevents invasion through niche pre-emption. By contrast, genotypic richness exerted a hump-shaped effect on invasion and was linked to the production of toxins antagonistic to the invader. This effect peaked at intermediate richness, suggesting that high richness levels may increase invasibility. Invasibility could be well predicted by the combination of these two mechanisms, documenting that both genotypic richness and dissimilarity need to be considered, if we are to understand the biotic properties determining the susceptibility of ecosystems to biological invasions.

Chemical and microbial properties in a greenhouse loamy soil after steam disinfestation alone or combined with CaO addition

Steam disinfestation represents an increasingly attractive strategy to control soil-borne pathogens and weeds both in greenhouses and field crops. Beyond pest control, steam injection has the potential to alter soil nutrient dynamics and composition of bacterial communities. This study investigated the impact of a soil-steaming method (the Bioflash™ system) on main chemical (total organic C, TOC; total N, TN; cation exchange capacity, CEC; exchangeable Na, K, Mg and Ca; KCl-extractable ammonium and nitrate, DTPA-exchangeable Mn) and microbial (microbial biomass C, MBC; genetic structure of bacterial communities) properties in a greenhouse loamy soil. Treatments were carried out by using a self-propelled soil-steaming machine operating at two steaming modes (deep or shallow) in combination or not with addition of an exothermically reacting compound (CaO at a 1000 kg ha −1 rate). Soils from a five-treatment (NS, non-steamed soil, DS, deep steam injection; DS + E, deep steam injection plus CaO; SS, shallow steam injection; SS + E, shallow steam injection plus CaO) plot block were sampled before steaming and after 8, 19, 54 and 91 days. Compositional shifts in the structure of soil bacterial communities were monitored by denaturing gradient gel electrophoresis (DGGE) fingerprinting of soil-extracted 16S rRNA gene fragments, using primers specific for Bacteria and Actinobacteria. Maximum soil temperature reached 70 °C (DS) and 82 °C (DS + E) in the 10–20 cm layer; and 50 °C (SS) and 53 °C (SS + E) in the 0–10 cm layer. TOC, TN, CEC, exchangeable Na, Mg and Ca were not statistically affected by soil steaming, CaO addition, depth of injection or their interactions. Conversely, SS promoted a large release of exchangeable K, notwithstanding CaO addition. Steam disinfestation significantly stimulated ammonium release, which further increased in CaO-treated steamed soils. Generally ammonium and nitrate pools varied oppositely, being the latter markedly reduced at the end of the observation period. Available Mn was significantly increased by soil steaming, but it was depressed when steaming was combined with CaO addition. Soil steaming markedly reduced the MBC, especially in DS-treated and SS-treated plots. The genetic structure of soil bacterial and actinobacterial communities was largely unresponsive to the steaming treatments. Even though the Bioflash™ system did not cause bacterial eradication or compositional shifts in bacterial community structure, soil steaming stimulated an increased release of soluble nutrients (K +, Mn 2+, NH 4 +–N). Over a longer perspective, excessive soil resources exploitation and potential risks of Mn toxicity should be cautiously considered especially in repeatedly (and eventually acid) steamed soils.

Bacteria diversity and microbial biomass in forest, pasture and fallow soils in the southwestern Amazon basin

It is well-known that Amazon tropical forest soils contain high microbial biodiversity. However, anthropogenic actions of slash and burn, mainly for pasture establishment, induce profound changes in the well-balanced biogeochemical cycles. After a few years the grass yield usually declines, the pasture is abandoned and is transformed into a secondary vegetation called "capoeira" or fallow. The aim of this study was to examine how the clearing of Amazon rainforest for pasture affects: (1) the diversity of the Bacteria domain evaluated by Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis (PCR-DGGE), (2) microbial biomass and some soil chemical properties (pH, moisture, P, K, Ca, Mg, Al, H + Al, and BS), and (3) the influence of environmental variables on the genetic structure of bacterial community. In the pasture soil, total carbon (C) was between 30 to 42 % higher than in the fallow, and almost 47 % higher than in the forest soil over a year. The same pattern was observed for N. Microbial biomass in the pasture was about 38 and 26 % higher than at fallow and forest sites, respectively, in the rainy season. DGGE profiling revealed a lower number of bands per area in the dry season, but differences in the structure of bacterial communities among sites were better defined than in the wet season. The bacterial DNA fingerprints in the forest were stronger related to Al content and the Cmic:Ctot and Nmic:Ntot ratios. For pasture and fallow sites, the structure of the Bacteria domain was more associated with pH, sum of bases, moisture, total C and N and the microbial biomass. In general microbial biomass in the soils was influenced by total C and N, which were associated with the Bacteria domain, since the bacterial community is a component and active fraction of the microbial biomass. Results show that the genetic composition of bacterial communities in Amazonian soils changed along the sequence forest-pasture-fallow.

Bacillus amyloliquefaciens BNM122, a potential microbial biocontrol agent applied on soybean seeds, causes a minor impact on rhizosphere and soil microbial communities

The increase in soybean productivity has contributed to a greater use of agrochemicals, which cause major problems, such as soil and water pollution and reduction of biodiversity, and have a negative impact on non-target species. The development of microbial biocontrol agents for soybean diseases can help to reduce pesticide abuse. Bacillus amyloliquefaciens BNM122 is a potential microbial biocontrol agent able to control the damping-off caused by Rhizoctonia solani when inoculated in soybean seeds, both in a plant growth chamber and in a greenhouse. In this study, we report the effect of soybean seed treatments with strain BNM122 or with two fungicides (thiram and carbendazim) on the structure and function of the bacterial community that colonizes the soybean rhizosphere. Also, soybean root nodulation by Bradyrhizobium japonicum, mycorrhization by arbuscular mycorrhizal fungi and plant growth were evaluated. We used the r- and K-strategist concept to evaluate the ecophysiological structure of the culturable bacterial community, community-level physiological profiles (CLPP) in Biolog™ EcoPlates to study bacterial functionality, and the patterns of 16S RNA genes amplified by PCR and separated by denaturing gradient gel electrophoresis (PCR–DGGE) to assess the genetic structure of the bacterial community. Neither the ecophysiological structure nor the physiological profiles of the soybean rhizosphere bacterial community showed important changes after seed inoculation with strain BNM122. On the contrary, seed treatment with fungicides increased the proportions of r-strategists and altered the metabolic profiles of the rhizosphere culturable bacterial community. The genetic structure of the rhizosphere bacterial community did not show perceptible changes between treated and non-treated seeds. Regarding the bacterial and fungal symbioses, seed treatments did not affect soybean nodulation, whereas soybean mycorrhization significantly decreased ( P < 0.05) in plants obtained from seeds treated with strain BNM122 or with the fungicides. However, a higher negative effect was observed in plants which seeds were treated with the fungicides. Plant growth was not affected by seed treatments. It can be concluded that soybean seed treatment with B. amyloliquefaciens BNM122 had a lesser effect on soil microbial community than that with the fungicides, and that these differences may be attributed to the less environmental persistence and toxic effects of the strain, which deserve further studies in order to develop commercial formulations.

Impact of a new biopesticide produced by Paenibacillus sp. strain B2 on the genetic structure and density of soil bacterial communities

The effect of paenimyxin, a new biopesticide produced by Paenibacillus sp. strain B2, on the density of soil bacterial communities was assessed by colony counting and by 16S rDNA and nirK quantitative polymerase chain reaction (PCR). Paenimyxin had a negative effect on the bacterial colony-forming unit (CFU) number, which was significantly reduced 2 and 4 days after treatment. The effect of paenimyxin on cultivatable bacteria was negligible 7 days after treatment. Approximately 10(7) 16S rDNA sequences per gram of soil (dry weight) were detected by quantitative PCR in all samples. Paenimyxin did not affect the quantification of 16S rDNA or of the denitrifying bacterial community. In addition, RISA fingerprinting showed that the genetic structure of the bacterial communities was significantly modified 2 days after paenimyxin application at 50 microM and 4 days after treatment at lower concentrations (0.5 and 5 microM). The impact of paenimyxin treatment on the genetic structure of soil bacterial communities was transient, as no effect could be observed after 7, 14 and 28 days when compared with the untreated control.

Dynamics and identification of soil microbial populations actively assimilating carbon from 13C‐labelled wheat residue as estimated by DNA‐ and RNA‐SIP techniques

This work is the first report on the use of DNA-, RNA-SIP approaches to elucidate the dynamics and the diversity of bacterial populations actively assimilating C derived from plant residues labelled at more than 90% (13)C. Wheat-residues, were incorporated and incubated into soil microcosms for 28 days. At the end of the incubation time, no more than 55% of the total CO(2) released was (13)C-labelled, suggesting the occurrence of an important priming effect process. After 7 days, more than 30% of the whole DNA extracted were labelled, allowing an efficient separation of labelled from unlabelled DNA using density gradient centrifugation. The genetic structure of bacterial community, assessed by Automated Ribosomal Intergenic Spacer Analysis technique, was deduced from the (13)C- and (12)C-fractions of control and enriched conditions, over the time course of the experiment. Dynamics showed that wheat residues directly induced a rapid and durable stimulation of fresh organic matter (FOM) degrading populations ((13)C), while specific soil organic matter (SOM) degrading populations ((12)C) seemed to be indirectly stimulated only at the early time point (t7d). After 14 days of incubations, 16S rRNA clone libraries were elaborated on (12)C- and (13)C-RNA extracted from enriched microcosms, as well as (12)C-RNA extracted from control condition. Stimulation of the beta- and gamma-subgroups of proteobacteria, where numerous populations were previously described as r-strategists or copiotrophic organisms, was recorded in the (13)C-fraction. In the mean time, several phyla like Actinobacteria, Cyanobacteria, Candidate, Gemmatimonadetes and Planctomycetes were only present in (12)C fractions. Surprisingly, several sequences affiliated to species characterized as oligotrophic organisms were retrieved in both types of fraction. Trophic relationships between soil bacteria involved in FOM and SOM degradation were discussed on the basis of different hypotheses of Fontaine and colleagues (2003) concerning the mechanisms of the priming effect induction.

Does disturbance and restoration of alpine grassland soils affect the genetic structure and diversity of bacterial and N2‐fixing populations?

Responses of bacterial communities to disturbance and restoration processes were investigated on alpine grassland soil. Bulk soil, rhizosphere soil and two soil separates, i.e. sand-size (2000-200 microm) and silt-size (50-2 microm) were sampled from undisturbed grassland soil to soil under restoration for 1 month, 1 year, 4 years and 13 years after disturbance. Automated ribosomal intergenic spacer analysis (ARISA) and restriction fragment length polymorphism (RFLP) of nifH gene pools were used to assay genetic structure of the bacterial communities and N2-fixing guild. According to the distribution of ARISA band length in bacterial phyla, the dominance of ARISA bands below 400 bp showed that Gram-positive bacteria would be predominant in the studied grassland soil when not disturbed. Disturbance affected the genetic structure of bacterial community and of N2-fixing guild in relation to their location within the selected habitats. Shifts in IGS and nifH profiles of bulk soil metagenome were larger than those observed from sand-size- and silt-size-fractions, accounting for 40-50% of the variance in the profiles. Restoration of the genetic structure of telluric bacteria community and N2-fixing populations was found to be influenced by the spatial heterogeneity of the soil and niche diversification. Particular bacterial genetic structure within distinct habitats were evidenced and must be defined as subdivisions of the meta-community of bulk soil. Scale of soil microbial diversity/stability relationships is discussed with special attention to disconnected bacterial habitat compared with whole soil with multiple niches.

2,4-D impact on bacterial communities, and the activity and genetic potential of 2,4-D degrading communities in soil

The key role of telluric microorganisms in pesticide degradation is well recognized but the possible relationships between the biodiversity of soil microbial communities and their functions still remain poorly documented. If microorganisms influence the fate of pesticides, pesticide application may reciprocally affect soil microorganisms. The objective of our work was to estimate the impact of 2,4-D application on the genetic structure of bacterial communities and the 2,4-D-degrading genetic potential in relation to 2,4-D mineralization. Experiments combined isotope measurements with molecular analyses. The impact of 2,4-D on soil bacterial populations was followed with ribosomal intergenic spacer analysis. The 2,4-D degrading genetic potential was estimated by real-time PCR targeted on tfdA sequences coding an enzyme specifically involved in 2,4-D mineralization. The genetic structure of bacterial communities was significantly modified in response to 2,4-D application, but only during the intense phase of 2,4-D biodegradation. This effect disappeared 7 days after the treatment. The 2,4-D degrading genetic potential increased rapidly following 2,4-D application. There was a concomitant increase between the tfdA copy number and the 14C microbial biomass. The maximum of tfdA sequences corresponded to the maximum rate of 2,4-D mineralization. In this soil, 2,4-D degrading microbial communities seem preferentially to use the tfd pathway to degrade 2,4-D.

Impact of feeding by Arenicola marina (L.) and ageing of faecal material on fatty acid distribution and bacterial community structure in marine sediments: An experimental approach

The fate of ingested eukaryotic photoautotrophic fatty acids during gut transit in the lugworm Arenicola marina (L.) and the influence of A. marina's faeces on the evolution of fatty acid distribution and bacterial community structure in superficial sediments were studied under laboratory conditions. Dead phytoplanktonic cells (food portions) were fed to individual A. marina and subsequently incubated, or allowed to directly incubate in the presence of fresh egesta or non-ingested sediment. Changes in fatty acid composition and genetic structure of bacterial communities during gut transit and/or incubation were monitored using gas-chromatography/mass-spectrometry and a DNA fingerprint approach (RISA), respectively. Results, supported by principal component analyses, suggest that A. marina's feeding activity can directly and indirectly affect the lipid biomarker composition and the bacterial community structure of inhabited sediments. Faecal casts produced from food portions appeared qualitatively enriched in saturated fatty acids relative to (poly)unsaturated ones due, partly, to an increase of some bacterial fatty acids and to the preferential removal of some polyunsaturated fatty acids (PUFAs). The incubation of food portions in the presence of fresh A. marina's egesta (designed to study the indirect impact of feeding by A. marina) induced a significant increase in the concentrations of C 20 and C 22 polyunsaturated fatty acids (PUFAs), whereas these compounds almost disappeared following direct feeding and subsequent incubation, indicating that some dietary fatty acids may be more accessible to biodegradation following passage through the gut of A. marina. The aforementioned increase in PUFAs was attributed to a bacterial production during incubation, suggesting the presence of PUFA-producing bacteria in the fresh egesta of A. marina. Those bacteria were either enteric bacteria that were released with the egesta or ingested bacteria that have survived gut passage, as suggested by the variations of the bacterial community structure (i.e. RISA profiles) during incubation. The results suggest that aged faeces from A. marina might be, in some circumstances, of relatively high nutritional value to trophic levels which are unable to synthesize essential PUFAs de novo. The presence of PUFA-producing bacteria in guts of marine lugworms deserves further attention.

Ranking the magnitude of crop and farming system effects on soil microbial biomass and genetic structure of bacterial communities

Biological soil characteristics such as microbial biomass, community structures, activities, and functions may provide important information on environmental and anthropogenic influences on agricultural soils. Diagnostic tools and detailed statistical approaches need to be developed for a reliable evaluation of these parameters, in order to allow classification and quantification of the magnitude of such effects. The DOK long-term agricultural field experiment was initiated in 1978 in Switzerland for the evaluation of organic and conventional farming practices. It includes three representative Swiss farming systems with biodynamic, bio-organic and conventional fertilization and plant protection schemes along with minerally fertilized and unfertilized controls. Effects on microbial soil characteristics induced by the long-term management at two different stages in the crop rotation, i.e. winter wheat after potato or corn, were investigated by analyzing soil bacterial community structures using analysis of PCR-amplified rRNA genes by terminal restriction fragment length polymorphism and ribosomal intergenic spacer analysis. Application of farmyard manure consistently revealed the strongest influence on bacterial community structures and biomass contents. Effects of management and plant protection regimes occurred on an intermediate level, while the two stages in the crop rotation had a marginal influence that was not significant.

Dynamic of the genetic structure of bacterial and fungal communities at different developmental stages of Medicago truncatula Gaertn. cv. Jemalong line J5

The genetic structure of bacterial and fungal communities was characterized in the rhizosphere of Medicago truncatula Gaertn. cv. Jemalong line J5 at five developmental stages (three vegetative and two reproductive stages), and in three compartments (bulk soil, rhizosphere soil and root tissues). The genetic structure of microbial communities was determined by cultivation-independent methods using directly extracted DNA that was characterized by automated ribosomal intergenic spacer analysis (ARISA). Principal component analyses (PCA) indicate that, for all developmental stages, the genetic structure of microbial communities differed significantly by compartment, with a major shift in the community in root tissues corresponding to the most intimate compartment with the plant. Differences were also recorded during plant development, the most significant being observed during the transition between vegetative and reproductive stages. Throughout this period, plants were shown to establish the highest level of symbiotic association (mycorrhization, nodulation) with arbuscular mycorrhizal fungi and Rhizobia. During the reproductive stages, the dynamics of the genetic structure differed between bacterial and fungal communities. At the last reproductive stage, the genetic structure of bacterial communities became close to that recorded during the first vegetative stages, suggesting a resilience phenomenon, whereas the genetic structure of fungal communities remained different from the vegetative stages and also from the early reproductive stages, suggesting a persistence of the rhizosphere effect.