Littérature scientifique sur le sujet « Soil PLFA analysis »

ABSTRACT Microorganisms that oxidize atmospheric methane in soils were characterized by radioactive labelling with14CH4 followed by analysis of radiolabelled phospholipid ester-linked fatty acids (14C-PLFAs). The radioactive fingerprinting technique was used to compare active methanotrophs in soil samples from Greenland, Denmark, the United States, and Brazil. The 14C-PLFA fingerprints indicated that closely related methanotrophic bacteria were responsible for the oxidation of atmospheric methane in the soils. Significant amounts of labelled PLFAs produced by the unknown soil methanotrophs coeluted with a group of fatty acids that included i17:0, a17:0, and 17:1ω8c (up to 9.0% of the total 14C-PLFAs). These PLFAs are not known to be significant constituents of methanotrophic bacteria. The major PLFAs of the soil methanotrophs (73.5 to 89.0% of the total PLFAs) coeluted with 18:1 and 18:0 fatty acids (e.g., 18:1ω9, 18:1ω7, and 18:0). The 14C-PLFAs fingerprints of the soil methanotrophs that oxidized atmospheric methane did not change after long-term methane enrichment at 170 ppm CH4. The 14C-PLFA fingerprints of the soil methanotrophs were different from the PLFA profiles of type I and type II methanotrophic bacteria described previously. Some similarity at the PLFA level was observed between the unknown soil methanotrophs and the PLFA phenotype of the type II methanotrophs. Methanotrophs in Arctic, temperate, and tropical regions assimilated between 20 and 54% of the atmospheric methane that was metabolized. The lowest relative assimilation (percent) was observed for methanotrophs in agricultural soil, whereas the highest assimilation was observed for methanotrophs in rain forest soil. The results suggest that methanotrophs with relatively high carbon conversion efficiencies and very similar PLFA compositions dominate atmospheric methane metabolism in different soils. The characteristics of the methane metabolism and the 14C-PLFA fingerprints excluded any significant role of autotrophic ammonia oxidizers in the metabolism of atmospheric methane.

ABSTRACT Phenanthrene-metabolizing soil microbial communities were characterized by examining mineralization of [14C]phenanthrene, by most-probable-number (MPN) counting, by 16S-23S spacer DNA analysis of the numerically dominant, culturable phenanthrene-degrading isolates, and by examining incorporation of [13C]phenanthrene-derived carbon into sterols and polar lipid fatty acids (PLFAs). An unpolluted agricultural soil, a roadside soil diffusely polluted with polycyclic aromatic hydrocarbons (PAHs), and two highly PAH-polluted soils from industrial sites were analyzed. Microbial phenanthrene degraders were not detected by MPN counting in the agricultural soil and the roadside soil. In the industrial soils, phenanthrene degraders constituted 0.04 and 3.6% of the total number of CFU. 16S-23S spacer DNA analysis followed by partial 16S DNA sequencing of representative isolates from one of the industrial soils showed that one-half of the isolates belonged to the genus Sphingomonas and the other half were closely related to an unclassified beta-proteobacterium. The 13C-PLFA profiles of the two industrial soils were relatively similar and resembled the profiles of phenanthrene-degrading Sphingomonas reference strains and unclassified beta-proteobacterium isolates but did not match the profiles of Pseudomonas, Mycobacterium, or Nocardia reference strains. The 13C-PLFA profiles of phenanthrene degraders in the agricultural soil and the roadside soil were different from each other and different from the profiles of the highly polluted industrial soils. Only in the roadside soil were 10me/12me18:0 PLFAs enriched in 13C, suggesting that actinomycetes metabolized phenanthrene in this soil. The 13C-PLFA profiles of the unpolluted agricultural soil did not resemble the profiles of any of the reference strains. In all of the soils investigated, no excess 13C was recovered in the 18:2ω6,9 PLFA, suggesting that fungi did not contribute significantly to assimilation of [13C]phenanthrene.

Abstract We studied soil PLFAs composition and specific soil properties among transect of small-scale fen in Stankovany, Slovakia. The aim of this study was to determine potential differences in the microbial community structure of the fen transect and reveal correlations among PLFAs and specific soil characteristics. PCA analyses of 43 PLFAs showed a separation of the samples along the axis largely influenced by i14:0, 16:1ω5, br17:0, 10Me16:0, cy17:0, cy17:1, br18:0 and 10Me17:0. We measured a high correlation of sample scores and distance from fen edge (Kendall’s test τ = 0.857, P < 0.01). Kendall’s test showed a negative correlation of PLFAs content (mol%) and distance from the fen border for Gram (+) bacteria, Actinomycetes, mid-chain branched saturated PLFAs and total PLFAs. The redundancy analysis of the PLFA data set for the eight samples using PLFAs as species and 21 environmental variables identified soil properties significantly associated with the PLFA variables, as tested by Monte Carlo permutation showing most significant environmental variables including dichlormethan extractables, water extractables, Klason lignin, acid-soluble lignin, holocellulose, total extractables, organic matter content, total PLFA amount, bacterial PLFA and total nitrogen negatively correlated to axis 1 and dry weight and carbonate carbon positively correlated to axis 1. The amounts of Klason lignin, acid-soluble lignin, holocellulose total extractables, total PLFA, bacterial PLFA and total nitrogen were significantly correlated positively to the distance from fen border while moisture and total carbonate carbon were correlated negatively.

Stable isotope probing of phospholipid fatty acids (PLFA-SIP) is useful when studying bacterial contributions to soil processes, and it is an effective way to separate fungal and bacterial activity by linking 13C enrichment to specific PLFAs. Distinguishing bacterial contributions to soil processes often employs selective inhibitors; however, studies demonstrating their efficacy when using PLFA-SIP are less common. Here, we determined the effect of the fungal inhibitor cycloheximide (4.8 mg g−1 dry soil) and the bacterial inhibitor bronopol (0.48 mg g−1 dry soil) on microbial communities white spruce [Picea glauca (Moench) Voss] forest floor by measuring the uptake of 13C-enriched glucose (2 mg g−1 dry soil) in microbial PLFAs. We targeted [13C]glucose uptake by the bacterial community conditioned to a stable soil environment of 23 °C for over 2 wk rather than new bacteria generated from active colony growth caused by glucose addition. Nearly all bacterial PLFAs exhibited pronounced inhibition of 13C enrichment in the presence of bronopol. Limited inhibition of 13C enrichment in the presence of cycloheximide was observed as bacterial PLFA affected by cycloheximide had roughly one third less 13C enrichment than samples emended with [13C]glucose alone. Inhibitory effects only reduced 13C enrichment and did not affect total PLFA concentrations, implying that the inhibitors in the concentrations applied were impeding bacterial activity without causing cell death. Based on this work, we conclude that bronopol is an effective inhibitor for bacteria. Additionally, non-targeted effects of cycloheximide on soil bacteria must be accounted for when it is used in soil incubations.

ABSTRACT Phospholipid fatty acid (PLFA) analysis of a soil microbial community was coupled with 13C isotope tracer analysis to measure the community’s response to addition of 35 μg of [13C]toluene ml of soil solution−1. After 119 h of incubation with toluene, 96% of the incorporated13C was detected in only 16 of the total 59 PLFAs (27%) extracted from the soil. Of the total 13C-enriched PLFAs, 85% were identical to the PLFAs contained in a toluene-metabolizing bacterium isolated from the same soil. In contrast, the majority of the soil PLFAs (91%) became labeled when the same soil was incubated with [13C]glucose. Our study showed that coupling13C tracer analysis with PLFA analysis is an effective technique for distinguishing a specific microbial population involved in metabolism of a labeled substrate in complex environments such as soil.

Under different soil salinity levels, diversity of soil microbial communities from Hetao irrigated land of Inner Mongolia was studied by phospholipid fatty acid (PLFA) analysis. The study found that PLFAs biomass in saline soil was significantly lower than those of strongly salinized soil and slight salinized soil. Microbes was bacteria-based from these soil. The bacterial PLFA loading in saline soil is significantly less than those of strongly salinized soil and slight salinized soil . Cluster analysis showed that changes had obviously taken place on soil microbial composition and quantity under different soil salinity levels.About 76.89% of variation in PLFA patterns explained by PC1(the first principal components),and 17:1, 16:0, 18:1w9c, 18:1w9t, 18:2, 18:3w3c, 12:0 were strongly negatively correlated with PC1.However,soil salinity and pH were positively correlated with PC1.We conclude that soil salinity has a profound affect on the microbial community structure.

A greenhouse pot test, in which wheat, cabbage, spinach were cultivated separately in petroleum contaminated soil with and without super absorbent polymer (SAP), was conducted to evaluated the effect of plants and SAP on soil microbiological properties. phospholipid fatty acids (PLFAs) profiles were analyzed to reveal the microbial communities. As a measure of the functional activity of soil microbial community, the ratio of degraded to total petroleum hydrocarbon (TPH) in soil was estimated. The results indicated that SAP had an important effect on the soil microbial community and its degrading TPH activities. First, the principal component analysis (PCA) of the PLFA signatures revealed marked changes between soil with SAP and without SAP. In addition, the total amount and the profile of PLFA were significantly different between the untreated and SAP-amended soils. Using PLFA patterns as a biomarker, it was found that gram-positive bacteria (G+) were more sensitive to SAP than gram-negative bacteria (G-), and the biomass of G+ was higher in soil with SAP than in that without SAP. Second, the crop could stimulate the growth of soil microorganisms; however, the differences depended clearly on the crop species. The G+ and G- biomass was increased in cabbage, spinach soil containing SAP, but was decreased in wheat soils. The population of fungi was increased in cabbage and spinach soils containing SAP, but was decreased in wheat soil with SAP. The population of actinomycetes was decreased in all soils with SAP. Third, the ratio of degraded to TPH was slightly increased in soil with SAP treatment, but a significant change depended on the crop species. In total, 12 different PLFAs were identified, including saturated, monounsaturated, branched, and polyunsaturated species. There was a clear difference in the PLFAs composition between soils with and without SAP.

A greenhouse pot experiment, in which 3 different crops (wheat, cabbage, spinach) were cultivated in soil with and without humics (HS), was conducted to evaluate the effect of HS on soil microbiological properties. Phospholipid fatty acid (PLFA) profiles were analyzed to reveal the microbial community structure. As a measure of the functional activity of soil microbial community, the ratio of degraded to total petroleum hydrocarbon in soil was estimated. The results indicated that HS had an important effect on the soil microbial community and its functional activities. First, the principal component analysis (PCA) of the PLFA signatures revealed marked changes between soil with HS and without HS. In addition, the total amount and the profile of PLFA were significantly different between the untreated and HS-amended soils. Using PLFA patterns as a biomarker, it was found that gram-positive bacteria (G+) were more sensitive to HS than gram-negative bacteria (G-), and the biomass of G+ was higher in soil with HS than in that without HS. Second, the crop could stimulate the growth of soil microorganisms; however, the differences depended clearly on the crop species. The G+ and G- biomass was increased in spinach soil containing HS, but was decreased in wheat and cabbage soils. The population of fungi was increased in wheat and spinach soils containing HS, but was decreased in cabbage soil. The population of actinomycetes was increased in cabbage soil, but was decreased in wheat and spinach soils containing HS. Third, the ratio of degraded to total petroleum hydrocarbon was also affected by the HS treatment. It was slightly increased in soil with HS treatment, but a significant change depended on the crop species. In general, fatty acids ranged from C13 to C19. In total, 25 different PLFAs were identified, including saturated (SAT), monounsaturated (MUFA), branched (BR), and polyunsaturated (PUFA) species. There was a clear difference in the PLFA composition between soils with and without HS.

Soil erosion is prevalent in karst areas, but few studies have compared the differences in the drivers for soil microbial communities among karst ecosystems with different soil depths, and most studies have focused on the local scale. To fill this research gap, we investigated the upper 20 cm soil layers of 10 shallow–soil depth (shallow–SDC, total soil depth less than 100 cm) and 11 deep–soil depth communities (deep–SDC, total soil depth more than 100 cm), covering a broad range of vegetation types, soils, and climates. The microbial community characteristics of both the shallow–SDC and deep–SDC soils were tested by phospholipid fatty acid (PLFAs) analysis, and the key drivers of the microbial communities were illustrated by forward selection and variance partitioning analysis. Our findings demonstrated that more abundant soil nutrients supported higher fungal PLFA in shallow–SDC than in deep–SDC (p < 0.05). Furthermore, stronger correlation between the microbial community and the plant–soil system was found in shallow–SDC: the pure plant effect explained the 43.2% of variance in microbial biomass and 57.8% of the variance in the ratio of Gram–positive bacteria to Gram–negative bacteria (G+/G−), and the ratio of fungi to total bacteria (F/B); the pure soil effect accounted for 68.6% variance in the microbial diversity. The ratio of microbial PLFA cyclopropyl to precursors (Cy/Pr) and the ratio of saturated PLFA to monounsaturated PLFA (S/M) as indicators of microbial stress were controlled by pH, but high pH was not conducive to microorganisms in this area. Meanwhile, Cy/Pr in all communities was >0.1, indicating that microorganisms were under environmental stress. Therefore, the further ecological restoration of degraded karst communities is needed to improve their microbial communities.

We investigated soil microbial community structure by phospholipid fatty acid (PLFA) analysis in a mature boreal spruce forest landscape in southern Norway, with low diversity of vascular plants. We investigated the spatial variation in PLFAs and the importance of environmental variables in 10 plots (each 13 samples) in a study area of 1 km × 1 km. The scales investigated were 15 cm to 10 m within study plots and 100 m to 1 km between study plots. Soil microbial biomass varied 10-fold and we found a large variation in microbial community structure, even at distances of 15 cm. Samples aggregated into plots when PLFAs were subjected to a principal components analysis. Plot identity explained 36.3% of the variation in the PLFAs and geostatistical analysis showed that the microbial community structure displayed spatial dependence at within-plot distances. Environmental variables differed significantly between all plots but explained only minor parts of the variation in the overall PLFA pattern. The vegetation variables were, however, the best at explaining the PLFA pattern, and up to 60% of within-plot variation in individual plots, respectively, could be explained by vegetation variables, pH, and soil depth.

Global climate change is becoming a central issue in contemporary science as well as politics. There is a long-lasting debate about the cause of the climate change: anthropogenic activity versus the natural cycle. However, a scientific consensus is coming a conclusion that the contemporary climate change is mainly caused by anthropogenic emissions of the greenhouse gases (GHG), including carbon dioxide (CO2), nitrous oxide (N2O) and methane (CH4). The main objective of the thesis is the monitoring of such GHG emissions from two ecosystem types: a forest and a rice paddy ecosystem. The forest site is a EMEP experimental station, taking part of the activity of GHG-AGOLU of FP7-JRC project, while the agricultural ecosystem was included in the CarboEurope project and represents also a Level 3 site in the frame of NitroEurope project. The gas monitoring was carried out in 2008. The thesis is composed by 4 chapters, corresponding to specific objectives. The first chapter is relative to the study of the spatial variability of the main soil chemical and physical properties on the basis of which the gas monitoring points were selected. The second and the third chapters are relative to a cropland site. In particular, the second chapter includes monitoring data of CH4, N2O and CO2 fluxes from the paddy field, both during the crop growth season and the fallow period, and the validation results of the DeNitrification DeComposition (DNDC) model, a process-oriented biogeochemical model used for simulating soil gas emissions from the paddy field, are reported. The third chapter contains the study of characterization of microbial community composition using phospholipid fatty acid analysis (PLFA), at eight sampling dates representative of different soil conditions and crop stages and consequently characterized by distinct soil greenhouse emission rates. The fourth and last chapter includes the monitoring study of soil respiration in a forest site and its partitioning into autotrophic and heterotrophic components, applying the indirect linear regression method.

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This thesis is part of a multiple scientific effort seeking to support the replacement of degraded brazilian pastures by systems which integrate different land use types such as crop, pasture, and forest plantation (collectively known as iCLF systems). Here, the focus was also to discuss the potentialities of an unusual statistical multivariate approach called ?Procrustes Analysis? in the plant and soil ecology framework. The current thesis has three chapters through which details of the Procrustes analysis are presented on both technically e intuitively manner. The first chapter describes roadmaps showing how the procrustean residual vector (so-called PAM: Procrustean association metric), representing the multivariate correlation between two or more data tables, can be used as an univariate variable in more user-traditional statistical approaches such as ecological ordination, regression analysis and ANOVA followed by mean comparisons. The second chapter discussed a case study and had as the general objective to use PAMs, depicting the relationships between distance matrices from individual soil microbial structure (PLFA: Phospholipids Fatty Acid) and distance matrices form soil properties variables (chemical and physic), as response variables in an ANOVA framework with land use type as categorical predictor (degraded pasture, improved pasture, native fragment and iCLF system). The hypothesis in this case was that the fungi:bacteria ratio given by PLFA analysis, a good index of changes in microbial structure as response to land use alteration and associated to more conservative soils in terms of carbon mineralization, is favored by the man ? introduced vegetal heterogeneity which characterizes the integration crop ? livestock ? forest. The last chapter was entirely dedicated to answer some technical questions which arose after the publication of the first chapters. Basically the two most common questions were: i) Does the increasing number of columns/variables within a data table affect Procrustes outcomes? ii) Can the procrustean residual vector, the PAM, translate differences between treatments in terms of multivariate correlation as it is used in mean comparisons? Specifically for these questions, Procrustes was useful in supporting iCLF systems as potential alternative to degraded pasture by raising insights that the man ? introduced vegetal heterogeneity in such integrated agroecosystem, favor shifts in microbial structure toward fungal dominance.
A presente tese fez parte do esfor?o multinstitucional buscando sustentar a substitui??o de pastagens degradas por sistemas que integrem diferentes tipos de uso da terra, mais especificamente aqueles integrando lavoura, pastagem, e floresta plantada, coletivamente: sistemas iLPF. Aqui, o foco foi a explora??o das potencialidades da abordagem estat?stica denominada an?lise Procrutes, ou simplesmente Procrustes, na seara de ecologia de planta e solo. Basicamente, a tese foi composta por tr?s cap?tulos onde ? descrito com detalhes os principais nuances dessa abordagem multivariada ainda pouco utilizada por ecologistas de planta e solo. O primeiro cap?tulo descreve roteiros esquem?ticos mostrando como o vetor de res?duos derivado da correla??o e duas tabelas de dados pela an?lise Procrustes (chamado PAM: Procrustes association metric) pode ser utilizado como representante univariado da correla??o em outras abordagens estat?sticas (ordena??o ecol?gica, regress?o, e ANOVA seguida de teste de m?dias). O segundo cap?tulo da tese, utilizando sugest?es do primeiro cap?tulo, tratou de um estudo de caso. Neste caso, fazenda experimental situada no munic?pio de Cachoeira dourada ? GO, e contendo quatro diferentes tipos de uso da terra, dentre os quais um sistema iLPF, foi escolhida para a condu??o do estudo de caso. O objetivo geral foi acessar como correla??es, no formato de PAM, entre tabelas de dados representadas por vari?veis individuais de estrutura microbiana (dada por an?lise de lip?dios oriundos do solo; PLFA: Phospholipids Fatty Acid) e propriedades individuais de qu?mica e f?sica de solo, eram moduladas pelo tipo de uso da terra: pastagem degradada, pastagem melhorada, fragmento de mata nativa, e sistema iLPF. A hip?tese para o estudo de caso foi a de que a rela??o fungo: bact?ria, comumente associada a ambientes mais conservativos, era promovida pelo sistema iLPF uma vez que tais sistemas s?o caracterizados pelo aumento da heterogeneidade vegetal oriunda da sistematizada introdu??o de especies arb?reas em meio a pastagem. O terceiro e ?ltimo cap?tulo da tese foi estritamente dedicado a responder questionamentos t?cnicos referentes ? abordagem procrusteana e surgidos depois das publica??es dos dois primeiros cap?tulos da tese. Neste caso, dois dos questionamentos mais comuns foram abordados. Foram eles: i) quais s?o os efeitos da correla??o entre colunas/vari?veis dentro de uma tabela de dados sobre os resultados da an?lise Procrustes? ii) Pode o vetor de res?duos procrusteanos, a PAM, traduzir diferen?as entre tratamentos em termos da for?a de correla??o multivariada entre duas tabelas de dados? Para o estudo de caso os resultados da corrente tese suportaram os sistemas iLPF como potencial alternativa para substitui??o de pastagens degradadas ao levantar ind?cios de que a heterogeneidade vegetal introduzida nos sistemas iLPF pode favorecer o deslocamento da estrutura microbiana em dire??o ao dom?nio de fungos.

Nowadays, plastic waste poses one of the greatest risks to the environment. Plastics affect the environment at all stages of their life cycle. Bioplastics have become widely used as a substitute for conventional plastics, without detailed examination of their behavior in real environmental conditions. As a result, it is assumed that they can accumulate in the environment and the question arose as to how to identify them. The main goal of this thesis is to develop a method based on sample pyrolysis that is suitable for the identification and determination of the amount of PLA microplastics in soil and other solid matrices. Three types of soils and sludge were used for analysis. These matrices were spiked to obtain concentration ranges 0,2% - 5,0%. The pyrolysis resulted in evolution of gases with the signals m/z 29, 43 and 44, witch originated from PLA and are suitable for qualitative and quantitative analysis. Analysis of PLA in sludge was more complicated due to similarity of gases evolved from pure matrices. We tested three approaches based on analysis of signal´s peak areas, intensities and temperatures of gas evolution. While the first approach failed, the last two approaches appeared to be promising for qualitative and quantitative analysis of PLA in the sludge. Several methods suitable for qualitative and quantitative analysis of even very small amounts of PLA in soils and sludge have also been designed/developed. These methods were based on analysis of the composition and dynamics of the released gases and the characterisctic degradation temperatures.

A Floresta Ombrófila Mista também chamada de Floresta de Araucária representa um dos mais ricos remanescentes de florestas pluviais subtropicais brasileiras, tendo como principal representante a Araucaria angustifolia, espécie considerada ameaçada de extinção. A diversidade microbiana possui um importante papel no funcionamento e manutenção do equilíbrio dos ecossistemas florestais, mas é desconhecida em solos com araucária. O presente estudo teve como objetivo avaliar a diversidade, funcionalidade e estrutura das comunidades microbianas em florestas de Araucaria angustifolia natural, introduzida e impactada pela queima acidental. O estudo foi realizado no Parque Estadual de Campos de Jordão, SP, tendo como áreas de estudo: floresta nativa com predomínio de araucária (FN), reflorestamento de araucária (RF), e reflorestamento de araucária com queima acidental em julho de 2001 (RQ). Em cada área, foram escolhidas ao acaso dez árvores de araucária e ao redor de cada árvore coletou-se uma amostra composta de três subamostras. Foram avaliados atributos químicos, microbiológicos, estrutura das comunidades de Bacteria e Archaea por PCR-DGGE e seqüenciamento parcial do gene rRNA 16S de Bacteria, análise da capacidade de utilização de substratos de carbono (Biolog) e perfis de ácidos graxos ligados a ésteres de fosfolipídios (PLFAs). As áreas são caracterizadas por solos ácidos, com elevado conteúdo de matéria orgânica (MO) e baixa disponibilidade de cátions metálicos básicos. A área FN apresentou maiores teores de carbono da biomassa microbiana (CBM), atividade respiratória basal (C-CO2) e relação carbono da biomassa microbiana: carbono orgânico total (CBM:COT), comparada ao RF e RQ. Os maiores valores de quociente metabólico (qCO2) foram encontrados no RQ quando comparado a FN e RF. A análise canônica discriminante identificou o atributo microbiano qCO2 e químicos, Mg e pH, como sendo responsáveis pela discriminação das áreas, seguidos do teor de P. A análise de PCRDGGE revelou que as estruturas das comunidades bacterianas de FN e RQ foram mais similares entre si do que em RF. A análise de escala multidimensional (NMDS) com ANOSIM baseada nos perfis de amplicons de Bacteria mostrou que as três áreas são diferentes entre si, enquanto para Archaea não houve diferença entre as áreas estudadas. A afiliação taxonômica das seqüências de clones do gene rRNA 16S mostrou que o solo de FN apresenta uma maior diversidade de táxons. Os filos Proteobacteria e Actinobacteria foram os mais freqüentes nas três áreas. A maior diversidade estimada pelo índice de Shannon foi encontrada em RQ, em comparação a FN e RF. A análise por Biolog mostrou que área FN apresenta a maior taxa de utilização de substratos, em relação RF e RQ, as quais não diferiram entre si. Os perfis de PLFAs não apresentaram diferenças entre as áreas estudadas. Observou-se uma maior biomassa bacteriana, principalmente de Gram-positivas, quando comparada a biomassa fúngica de PLFAs das áreas estudadas. As análises multivariadas apresentaram-se como importantes ferramentas no estudo de diversidade microbiana, sendo os atributos químicos e microbiológicos do solo úteis para a interpretação dos resultados obtidos.
The Ombrophilic Mixed Forest, also called Araucaria Forest, represents one of the richest remainders of subtropical pluvial forests in Brazil. Its main representative species is the endangered Araucaria angustifolia. The microbial diversity plays an important role in functioning of forest ecosystems. However, the microbial diversity in soils with araucaria forests is mostly unknown. The aim of this work was to evaluate the diversity, structure of microbial communities in their possible functions in a natural preserved araucaria forest (FN), a planted araucaria forest (RF) and a planted araucaria forest impacted by accidental fire (RQ). The study was carried out at the State Park of Campos of Jordão (SP). For each area, ten araucaria trees were randomly selected and a sample composed of three sub-samples was collected at approximately one meter from the trunk of each tree. Chemical and microbiological attributes, as well as structures of bacterial and archaeal communities were evaluated using PCR-DGGE and the partial sequencing of the 16S rRNA gene from Bacteria, community level physiological profiles using Biolog, the phospholipid fatty acids profiles (PLFAs). The studied areas were characterized by acidic soils, with high content of organic matter (OM) and low availability of basic metallic cations. The area FN presented the highest contents of carbon in the microbial biomass (CBM), higher basal respiration activity (C-CO2) and higher microbial biomass carbon: total organic carbon ratio (CBM:TOC), compared to RF and RQ. The highest values of metabolic quotient (qCO2) were observed in RQ, when compared to FN and RF. Using canonical discriminant analysis (CDA), qCO2, Mg concentration and pH were identified as the main attributes responsible for the discrimination of the areas, followed by the P concentration. The PCR-DGGE analysis revealed that the bacterial community structures in FN and RQ share higher levels of similarity, as compared to RF. Non-metric multidimensional scale analysis (NMDS) and ANOSIM based on the profiles of bacterial 16S rRNA gene amplicons showed that the all three areas had different bacterial communities, whereas archaeal communities were similar, based on 16S rRNA genes amplicon profiles. The phylogenetic affiliation of 16S rRNA gene clone sequences showed that soil from FN presents higher taxa diversity, as compared to RF and RQ. The phyla Proteobacteria and Actinobacteria were the most frequent in the three areas studied. Higher Shannon index was observed in RQ soil than FN and RF soils. Biolog analysis showed that FN has the highest substrate utilization rates, when compared to RF and RQ, which did not show significant differences. In general, PLFAs profiles did not show differences for the areas studied. Estimated bacterial biomass was higher than fugal biomass, with predominance of Gram-positive bacteria. Integration of chemical and microbial attributes through multivariate analyses is essential for identifying the factors determining microbial community structure in forest soils.

The PLFA profiling method was adopted and used to determine changes in microbial community structure and abundance along natural and human-induced environmental gradients. The presented studies were based on field sampling campaigns combined with targeted laboratory experiments. According to the aims of particular studies, microbial PLFA fingerprinting was combined with the auxiliary below- and aboveground ecosystem characteristics to identify the drivers of microbial responses to environmental changes or with 13C-labelling and metagenomics to obtain more complex information about running processes and involved microorganisms.

Biochar is a charcoal like substance produced from organic biomass after pyrolysis. Biochar act as a good soil conditioner by increasing microbial activities, soil nutrition and soil structure. Soil microorganisms are involved in litter decomposition and soil nutrient mineralization which is important in the sustainable development of plants and trees. The functioning of an ecosystem is controlled by biogeochemical cycles driven by microorganisms. The cell membrane of all microorganisms is composed of phospholipids that are easily metabolized after the cell death. Hence, phospholipid fatty acid (PLFA) analysis of microorganisms can be used for the characterization of living microbial communities. PLFA analysis is a lipid based, culture independent biochemical technique. Therefore, PLFAs can be used for the characterization of soil microbial community structure that are not able to cultivated by the conventional methods. This profiling act as a biological register of soil health, and as an indicator of soil response to different field management systems like biochar.