Antagonism between Bacteria and Fungi on Decomposing Aquatic Plant Litter

Bacterial and fungal decomposers of aquatic plant litter may exhibit either synergistic or antagonistic interactions, which are likely to influence microbial growth as well as the decomposition of litter and, eventually, the carbon metabolism of aquatic systems. To elucidate such interactions, we inoculated decomposing Phragmites culms in microcosms with fungal isolates and with natural communities of bacteria and fungi in different combinations. The development of fungal and bacterial biomass and the carbon dynamics were studied during several months of degradation. The results show a bilateral antagonistic relationship between bacteria and fungi. After 3 months, fungal biomass accumulation was approximately 12 times higher in the absence than in the presence of bacteria. Bacterial biomass accumulation was about double in the absence of fungi compared to when fungi were present. Similar interactions developed between a natural assemblage of bacteria and five different fungal strains isolated from Phragmites litter (three identified hyphomycetes and two unidentified strains). Despite the great difference in biomass development between the treatments, the carbon metabolism was similar regardless of whether fungi and/or bacteria were present alone or in coexistence. We suggest that the antagonism between bacteria and fungi is an important controlling factor for microbial colonization and growth on aquatic plant litter.     

Identification of cellulose-responsive bacterial and fungal communities in geographically and edaphically different soils by using stable isotope probing

Many bacteria and fungi are known to degrade cellulose in culture, but their combined response to cellulose in different soils is unknown. Replicate soil microcosms amended with [(13)C]cellulose were used to identify bacterial and fungal communities responsive to cellulose in five geographically and edaphically different soils. The diversity and composition of the cellulose-responsive communities were assessed by DNA-stable isotope probing combined with Sanger sequencing of small-subunit and large-subunit rRNA genes for the bacterial and fungal communities, respectively. In each soil, the (13)C-enriched, cellulose-responsive communities were of distinct composition compared to the original soil community or (12)C-nonenriched communities. The composition of cellulose-responsive taxa, as identified by sequence operational taxonomic unit (OTU) similarity, differed in each soil. When OTUs were grouped at the bacterial order level, we found that members of the Burkholderiales, Caulobacteriales, Rhizobiales, Sphingobacteriales, Xanthomonadales, and the subdivision 1 Acidobacteria were prevalent in the (13)C-enriched DNA in at least three of the soils. The cellulose-responsive fungi were identified as members of the Trichocladium, Chaetomium, Dactylaria, and Arthrobotrys genera, along with two novel Ascomycota clusters, unique to one soil. Although similarities were identified in higher-level taxa among some soils, the composition of cellulose-responsive bacteria and fungi was generally unique to a certain soil type, suggesting a strong potential influence of multiple edaphic factors in shaping the community.     

Microbial decomposition of reed (Phragmites communis) leaves in a saline lake

Microbial colonization and its relation to the decomposition of reed (Phragmites communis) leaf litter were studied in the littoral area of a saline lake from autumn to summer using litter bag method. There was considerable fungal population on the leaves at the beginning of submergence. These fungi were probably terrestrial in origin. The fungal population rapidly disappeared few days after submergence, when bacteria, including cellulolytic and xylanolytic types, proliferated. Associated with this rapid colonization of bacteria, decomposition rates of cellulose and xylan increased. The rates declined from day 39 to day 100 with decreasing water temperature, though cellulolytic and xylanolytic bacteria maintained a sizeable population until day 150. A community of cellulolytic and xylanolytic fungi increased steeply after day 150. It coincided with a second increase in decomposition rate. These results suggest that the principal decomposers of reed leaf litter were bacteria in the initial phase and fungi in the later phase of the experiment.     

Eisenia fetida (Oligochaeta, Lumbricidae) Activates Fungal Growth, Triggering Cellulose Decomposition During Vermicomposting

Cellulose is the most abundant polymer in nature and constitutes a large pool of carbon for microorganisms, the main agents responsible for soil organic matter decomposition. Cellulolysis occurs as the result of the combined action of fungi and bacteria with different requirements. Earthworms influence decomposition indirectly by affecting microbial population structure and dynamics and also directly because the guts of some species possess cellulolytic activity. Here we assess whether the earthworm Eisenia fetida (Savigny 1826) digests cellulose directly (i.e., with its associated gut microbiota) and also whether the effects of E. fetida on microbial biomass and activity lead to a change in the equilibrium between fungi and bacteria. By enhancing fungal communities, E. fetida would presumably trigger more efficient cellulose decomposition. To evaluate the role of E. fetida in cellulose decomposition, we carried out an experiment in which pig slurry, a microbial-rich substrate, was treated in small-scale vermireactors with and without earthworms. The presence of earthworms in vermireactors significantly increased the rate of cellulose decomposition (0.43 and 0.26% cellulose loss day⁻¹, with and without earthworms, respectively). However, the direct contribution of E. fetida to degradation of cellulose was not significant, although its presence increased microbial biomass (C_mic) and enzyme activity (cellulase and β-glucosidase). Surprisingly, as fungi may be part of the diet of earthworms, the activity of E. fetida triggered fungal growth during vermicomposting. We suggest that this activation is a key step leading to more intense and efficient cellulolysis during vermicomposting of organic wastes.     

不同树叶凋落物对人参土壤理化性质及微生物群落结构的影响

树种选择是林下山参护育成败的关键,研究树叶凋落物对人参土壤养分、微生物群落结构组成的影响,旨在为林下山参护育选择适宜林地及农田栽参土壤改良提供科学依据和理论指导。通过盆栽试验,研究添加5.0 g色木槭Acer mono.Maxim.var.mono(A)、赤松Pinus densiflora Sieb.et Zucc.(B)、胡桃楸Juglans mandshurica Maxim.(C)、紫椴Tilia amurensis Rupr.(D)、蒙古栎Quercus mongolica Fisch.ex Ledeb.(E)树叶凋落物到土壤中,种植人参(Panax ginseng C.A.meyer)后研究土壤理化性质以及微生物群落结构的变化。结果表明:添加不同树叶处理后人参土壤性质发生改变,土壤p H值显著高于对照土壤5.91(P0.05),土壤全氮、速效氮磷、微生物碳氮在所有树叶处理中显著增加(P0.05),而土壤容重、速效钾和C/N在添加树叶处理中降低。18个土壤样品基因组,经16S和ITS1测序分别得到6064和1900个OUTs。其中细菌涵盖了42门、117纲、170目、213科、225属,真菌涵盖了24门、98纲、196目、330科、435属。不同树叶处理人参土壤细菌和真菌地位发生改变,细菌Proteobacteria是树叶分解的关键微生物,添加树叶后其多样性显著高于对照(P0.05)。而细菌Bacteroidetes和真菌Basidiomycota可能是区别阔叶林和针叶林树种的关键微生物,针叶林中含量显著低于阔叶林(P0.05),而真菌Ascomycota是针叶林分解的关键微生物。进一步从不同分类水平上得到特定树叶凋落物的特异细菌和真菌。典型相关分析(CDA)表明细菌Bacteroidetes、Chloroflexi、Actinobacteria及真菌Basidiomycota、Zygomycota、Chytridiomycota及Ascomycota的位置及多样性的改变均与土壤因子SMBN、TN、AP、SOC、AK、C/N、p H有关。综上所述,添加不同树叶后不仅提高土壤微生物量碳氮、改善土壤理化性质,同时改变微生物群落结构组成,不同树叶处理土壤理化性质不同导致人参土壤微生物组成的差异,本结果对于林下参选地和农田栽参土壤微生物改良具有理论指导作用。     

Effect of Inorganic Nutrients on Relative Contributions of Fungi and Bacteria to Carbon Flow from Submerged Decomposing Leaf Litter

The relative contributions of fungi and bacteria to carbon flow from submerged decaying plant litter at different levels of inorganic nutrients (N and P) were studied. We estimated leaf mass loss, fungal and bacterial biomass and production, and microbial respiration and constructed partial carbon budgets for red maple leaf disks precolonized in a stream and then incubated in laboratory microcosms at two levels of nutrients. Patterns of carbon flow for leaf disks colonized with the full microbial assemblage were compared with those colonized by bacteria but in which fungi were greatly reduced by placing leaf disks in colonization chambers sealed with membrane filters to exclude aquatic hyphomycete conidia but not bacterial cells. On leaves colonized by the full microbial assemblage, elevated nutrient concentrations stimulated fungi and bacteria to a similar degree. Peak fungal and bacterial biomass increased by factors of 3.9 and 4.0; cumulative production was 3.9 and 5.1 times higher in the high nutrient in comparison with the low nutrient treatment, respectively. Fungi dominated the total microbial biomass (98.4 to 99.8%) and cumulative production (97.3 and 96.5%), and the fungal yield coefficient exceeded that of bacteria by a factor of 36 and 27 in low- and high-nutrient treatments, respectively. Consequently, the dominant role of fungi in leaf decomposition did not change as a result of nutrient manipulation. Carbon budgets indicated that 8% of leaf carbon loss in the low-nutrient treatment and 17% in the high-nutrient treatment were channeled to microbial (essentially fungal) production. Nutrient enrichment had a positive effect on rate of leaf decomposition only in microcosms with full microbial assemblages. In treatments where fungal colonization was reduced, cumulative bacterial production did not change significantly at either nutrient level and leaf decomposition rate was negatively affected (high nutrients), suggesting that bacterial participation in carbon flow from decaying leaf litter is low regardless of the presence of fungi and nutrient availability. Moreover, 1.5 and 2.3 times higher yield coefficients of bacteria in the reduced fungal treatments at low and high nutrients, respectively (percentage of leaf carbon loss channeled to bacterial production), suggest that bacteria are subjected to strong competition with fungi for resources available in leaf litter.     

Isolation of fungal cellobiohydrolase I genes from sporocarps and forest soils by PCR

Cellulose is the major component of plant biomass, and microbial cellulose utilization is a key step in the decomposition of plant detritus. Despite this, little is known about the diversity of cellulolytic microbial communities in soil. Fungi are well known for their cellulolytic activity and mediate key functions during the decomposition of plant detritus in terrestrial ecosystems. We developed new oligonucleotide primers for fungal exocellulase genes (cellobiohydrolase, cbhI) and used these to isolate distinct cbhI homologues from four species of litter-decomposing basidiomycete fungi (Clitocybe nuda, Clitocybe gibba, Clitopilus prunulus, and Chlorophyllum molybdites) and two species of ascomycete fungi (Xylaria polymorpha and Sarcoscypha occidentalis). Evidence for cbhI gene families was found in three of the four basidiomycete species. Additionally, we isolated and cloned cbhI genes from the forest floor and mineral soil of two upland forests in northern lower Michigan, one dominated by oak (Quercus velutina, Q. alba) and the other dominated by sugar maple (Acer saccharum) and American basswood (Tilia americana). Phylogenetic analysis demonstrated that cellobiohydrolase genes recovered from the floor of both forests tended to cluster with Xylaria or in one of two unidentified groups, whereas cellobiohydrolase genes recovered from soil tended to cluster with Trichoderma, Alternaria, Eurotiales, and basidiomycete sequences. The ability to amplify a key fungal gene involved in plant litter decomposition has the potential to unlock the identity and dynamics of the cellulolytic fungal community in situ.     

Effects of grassland species on decomposition of litter and soil microbial communities

Decomposition of litter is greatly influenced not only by its chemical composition but also by activities of soil decomposers. By using leaf litter from 15 plant species collected from semi-natural and improved grasslands, we examined (1) how interspecific differences in the chemical composition of litter influence the abundance and composition of soil bacterial and fungal communities and (2) how such changes in microbial communities are related to the processes of decomposition. The litter from each species was incubated in soil of a standard composition for 60 days under controlled conditions. After incubation, the structure of bacterial and fungal communities in the soil was examined using phospholipid fatty-acid analysis and denaturing gradient gel electrophoresis. Species from improved grasslands had significantly higher rates of nitrogen mineralization and decomposition than those from semi-natural grasslands because the former were richer in nitrogen. Litter from improved grasslands was also richer in Gram-positive bacteria, whereas that from semi-natural grasslands was richer in actinomycetes and fungi. Nitrogen content of litter also influenced the composition of the fungal community. Changes in the composition of both bacterial and fungal communities were closely related to the rate of litter decomposition. These results suggest that plant species greatly influence litter decomposition not only through influencing the quality of substrate but also through changing the composition of soil microbial communities.     

Aerobic cellulolytic bacterial flora associated with decomposing Phragmites leaf litter in a seawater lake

A litter bag experiment was carried out in a eutrophic seawater lake from autumn to summer in order to determine which bacterial genera play an important role in decomposition of Phragmites communis leaf litter. The count of cellulolytic bacteria and decomposition rate of litter cellulose increased rapidly during the initial month. In contrast, the count of cellulolytic fungi was lowest in this period. Pseudomonas accounted for 65–90% of total isolates of cellulolytic bacteria up to 5 months. These results suggest that Pseudomonas plays an important role in at least the initial decomposition stage of the litter.     

Responses of soil cellulolytic fungal communities to elevated atmospheric CO₂ are complex and variable across five ecosystems

Elevated atmospheric CO(2) generally increases plant productivity and subsequently increases the availability of cellulose in soil to microbial decomposers. As key cellulose degraders, soil fungi are likely to be one of the most impacted and responsive microbial groups to elevated atmospheric CO(2). To investigate the impacts of ecosystem type and elevated atmospheric CO(2) on cellulolytic fungal communities, we sequenced 10,677 cbhI gene fragments encoding the catalytic subunit of cellobiohydrolase I, across five distinct terrestrial ecosystem experiments after a decade of exposure to elevated CO(2). The cbhI composition of each ecosystem was distinct, as supported by weighted Unifrac analyses (all P-values; < 0.001), with few operational taxonomic units (OTUs) being shared across ecosystems. Using a 114-member cbhI sequence database compiled from known fungi, less than 1% of the environmental sequences could be classified at the family level indicating that cellulolytic fungi in situ are likely dominated by novel fungi or known fungi that are not yet recognized as cellulose degraders. Shifts in fungal cbhI composition and richness that were correlated with elevated CO(2) exposure varied across the ecosystems. In aspen plantation and desert creosote bush soils, cbhI gene richness was significantly higher after exposure to elevated CO(2) (550 μmol mol(-1)) than under ambient CO(2) (360 μmol mol(-1) CO(2)). In contrast, while the richness was not altered, the relative abundance of dominant OTUs in desert soil crusts was significantly shifted. This suggests that responses are complex, vary across different ecosystems and, in at least one case, are OTU-specific. Collectively, our results document the complexity of cellulolytic fungal communities in multiple terrestrial ecosystems and the variability of their responses to long-term exposure to elevated atmospheric CO(2).     

Microbial growth and detritus transformations during decomposition of leaf litter in a stream

SUMMARY. 1. Despite the widely accepted importance of bacteria and fungi in degrading detritus in aquatic ecosystems there is still very little quantitative information on the abundance and dynamics of these microorganisms. Using epifluorescent microscopy, we measured the biomass of bacteria and fungi during decomposition of three types of leaf detritus. Bacterial production was determined from the rate of incorporation of ³H-thymidine into DNA.
2. The transformation of leaf carbon into dissolved organic carbon and fine particulate organic carbon was followed in order to compare the amounts of leaf material that were converted into these 'end-products' of decomposition versus the amount converted into microbial biomass.
3. The amount of microbial carbon in the leaf-detritus complex never exceeded 5.2% of the total carbon, and fungal biomass was always much greater than bacterial biomass. Despite the greater standing stock of fungi, the rapid turnover of bacteria (doubling about once per day) implies that their role in degrading leaf litter or as a food source for detritivores might be as great as for fungi.
4. Removal of microbial biomass from leaf litter may occur as release of fungal spores and consumption or shedding of bacterial biomass. Fungal spores can be a significant part of the fine particulate organic carbon released from leaf detritus and potentially represent an important food resource for filter-feeding organisms.     

Fungal lysis by a soil bacterium fermenting cellulose

Recycling of plant biomass by a community of bacteria and fungi is fundamental to carbon flow in terrestrial ecosystems. Here we report how the plant fermenting, soil bacterium Clostridium phytofermentans enhances growth on cellulose by simultaneously lysing and consuming model fungi from soil. We investigate the mechanism of fungal lysis to show that among the dozens of different glycoside hydrolases C. phytofermentans secretes on cellulose, the most highly expressed enzymes degrade fungi rather than plant substrates. These enzymes, the GH18 Cphy1799 and Cphy1800, synergize to hydrolyse chitin, a main component of the fungal cell wall. Purified enzymes inhibit fungal growth and mutants lacking either GH18 grow normally on cellulose and other plant substrates, but have a reduced ability to hydrolyse chitinous substrates and fungal hyphae. Thus, C. phytofermentans boosts growth on cellulose by lysing fungi with its most highly expressed hydrolases, highlighting the importance of fungal interactions to the ecology of cellulolytic bacteria.     

Microbial Dynamics Associated with Multiphasic Decomposition of 14C-Labeled Cellulose in Soil

Abstract Recent emphasis on residue management in sustainable agriculture highlights the importance of elucidating the mechanisms of microbial degradation of cellulose. Cellulose decomposition and its associated microbial dynamics in soil were investigated in incubation experiments. Population dynamics of actinomycetes, bacteria, and fungi were monitored by direct counts. Populations of oligotrophic bacteria in cellulose-amended soil were determined by plate count using a low C medium containing 4 mg C liter⁻¹ agar, and copiotrophs using a high C medium. Cumulative ¹⁴CO₂ evolution from ¹⁴C-labeled cellulose was best described by a multiphasic curve in a 28-day incubation experiment. The initial phase of decomposition was attributed mainly to the activity of bacterial populations with a low oligotroph-to-copiotroph ratio, and the second phase mainly to fungal populations. An increase in oligotroph-to-copiotroph ratio coincided with the emergence of a rapid ¹⁴CO₂ evolution stage. Streptomycin reduced ¹⁴CO₂ evolution during the first phase and prompted earlier emergence of the second phase, compared to the control. Cycloheximide initially promoted ¹⁴CO₂ evolution but subsequently had a lasting negative effect on ¹⁴CO₂ evolution. Cycloheximide addition significantly increased bacterial biomass and resulted in substantially stronger oscillation of active bacterial populations, whereas it initially reduced, and then stimulated, active fungal biomass. The observed changes in ¹⁴CO₂ evolution could not be explained by observed shifts in fungal and bacterial biomass, probably because functional groups of fungi and bacteria could not be distinguished. However, it was suggested that oligotrophic bacteria prompted activation of cellulolytic enzumes in fungi and played an important role in leading to fungal dominance of cellulose decomposition. Received: 2 October 1995; Accepted: 10 February 1996     

Fungal diversity within the phyllosphere of Pinus massoniana and the possible involvement of phyllospheric fungi in litter decomposition

Fungi play key roles in forest ecosystems and help to shape the forest’s diverse functions. However, little is known about the diversity of phyllospheric fungi or their possible relationships with fungal communities residing in different micro-environments of Pinus massoniana forests. We investigated seven different sample types: mature needles (NM), dead needles (ND), needles falling as litter (L), fermenting needles (F), humus (H), top soil (0–20 cm) (TS), and secondary soil (20–40 cm) (SS). These seven fungal communities were examined and compared with ITS amplicons using a high-throughput sequencing technique. A total of 1213 fungal operational taxonomic units (OTUs) were obtained at a 97% sequence similarity level. Distinct fungal communities were associated with different sample types. A greater number of OTUs were present in both NM and F samples than those shared by both NM and TS samples, indicating that phyllospheric fungi may play crucial roles in litter decomposition. Sixty OTUs (the core microbiome) were found in all sample types, and they may probably play different ecological roles in different sample types. These findings extend our knowledge of the fungal diversity of the phyllosphere and its possible interactions with fungal communities found in distinct forest micro-habitats.     

沙坡头地区凋落物分解及其对土壤微生物群落的影响

以腾格里沙漠东南缘沙坡头人工固沙植被区典型植物种凋落物(小画眉草、藓类、油蒿叶片)为对象,运用凋落物分解袋法和高通量测序技术,分析了3种植物凋落物分解特征及其对土壤微生物群落的影响。结果表明: 分解时间和凋落物类型均显著影响分解速率,藓类分解最慢,13个月后质量损失比仅为15.4%,油蒿叶片和小画眉草的平均分解速率分别是藓类的4.9和3.4倍。经过11个月的分解,细菌群落的优势菌门为放线菌门和变形菌门,真菌群落的优势菌门是子囊菌门;藓类分解过程中,拟杆菌门和绿弯菌门的相对丰度显著增加,担子菌门的相对丰度显著降低。凋落物分解后,细菌和真菌群落物种多样性和丰富度显著增加,细菌群落组成在凋落物间变化不显著,真菌群落变化显著。凋落物的分解速率与细菌和真菌群落多样性及丰富度均呈负线性变化。植物多糖、全磷和土壤pH、微生物生物量氮、铵态氮含量是影响微生物群落结构的主要因子。凋落物分解改变了土壤微生物群落物种组成和种间相似性,显著增加了土壤中微生物群落的多样性和丰富度,促进了土壤生境的恢复。     

Characterization of trapped lignin-degrading microbes in tropical forest soil

Lignin is often the most difficult portion of plant biomass to degrade, with fungi generally thought to dominate during late stage decomposition. Lignin in feedstock plant material represents a barrier to more efficient plant biomass conversion and can also hinder enzymatic access to cellulose, which is critical for biofuels production. Tropical rain forest soils in Puerto Rico are characterized by frequent anoxic conditions and fluctuating redox, suggesting the presence of lignin-degrading organisms and mechanisms that are different from known fungal decomposers and oxygen-dependent enzyme activities. We explored microbial lignin-degraders by burying bio-traps containing lignin-amended and unamended biosep beads in the soil for 1, 4, 13 and 30 weeks. At each time point, phenol oxidase and peroxidase enzyme activity was found to be elevated in the lignin-amended versus the unamended beads, while cellulolytic enzyme activities were significantly depressed in lignin-amended beads. Quantitative PCR of bacterial communities showed more bacterial colonization in the lignin-amended compared to the unamended beads after one and four weeks, suggesting that the lignin supported increased bacterial abundance. The microbial community was analyzed by small subunit 16S ribosomal RNA genes using microarray (PhyloChip) and by high-throughput amplicon pyrosequencing based on universal primers targeting bacterial, archaeal, and eukaryotic communities. Community trends were significantly affected by time and the presence of lignin on the beads. Lignin-amended beads have higher relative abundances of representatives from the phyla Actinobacteria, Firmicutes, Acidobacteria and Proteobacteria compared to unamended beads. This study suggests that in low and fluctuating redox soils, bacteria could play a role in anaerobic lignin decomposition.     

The microbial ecology of anaerobic cellulose degradation in municipal waste landfill sites: evidence of a role for fibrobacters

Cellulose is reputedly the most abundant organic polymer in the biosphere, yet despite the fundamental role of cellulolytic microorganisms in global carbon cycling and as potential sources of novel enzymes for biotechnology, their identity and ecology is not well established. Cellulose is a major component of landfill waste and its degradation is therefore a key feature of the anaerobic microbial decomposition process. Here, we targeted a number of taxa containing known cellulolytic anaerobes (members of the bacterial genus Fibrobacter, lineages of Clostridium clusters I, III, IV and XIV, and anaerobic fungi of the Neocallimastigales) in landfill leachate and colonized cellulose 'baits' via PCR and quantitative PCR (qPCR). Fibrobacter spp. and Clostridium clusters III, IV and XIV were detected in almost all leachate samples and cluster III and XIV clostridia were the most abundant (1-6% and 1-17% of total bacterial 16S rRNA gene copies respectively). Two landfill leachate microcosms were constructed to specifically assess those microbial communities that colonize and degrade cellulose substrates in situ. Scanning electron microscopy (SEM) of colonized cotton revealed extensive cellulose degradation in one microcosm, and Fibrobacter spp. and Clostridium cluster III represented 29% and 17%, respectively, of total bacterial 16S rRNA gene copies in the biofilm. Visible cellulose degradation was not observed in the second microcosm, and this correlated with negligible relative abundances of Clostridium cluster III and Fibrobacter spp. (≤ 0.1%), providing the first evidence that the novel fibrobacters recently detected in landfill sites and other non-gut environments colonize and degrade cellulose substrates in situ.     

Culture-Dependent and -Independent Methods Capture Different Microbial Community Fractions in Hydrocarbon-Contaminated Soils

Bioremediation is a cost-effective and sustainable approach for treating polluted soils, but our ability to improve on current bioremediation strategies depends on our ability to isolate microorganisms from these soils. Although culturing is widely used in bioremediation research and applications, it is unknown whether the composition of cultured isolates closely mirrors the indigenous microbial community from contaminated soils. To assess this, we paired culture-independent (454-pyrosequencing of total soil DNA) with culture-dependent (isolation using seven different growth media) techniques to analyse the bacterial and fungal communities from hydrocarbon-contaminated soils. Although bacterial and fungal rarefaction curves were saturated for both methods, only 2.4% and 8.2% of the bacterial and fungal OTUs, respectively, were shared between datasets. Isolated taxa increased the total recovered species richness by only 2% for bacteria and 5% for fungi. Interestingly, none of the bacteria that we isolated were representative of the major bacterial OTUs recovered by 454-pyrosequencing. Isolation of fungi was moderately more effective at capturing the dominant OTUs observed by culture-independent analysis, as 3 of 31 cultured fungal strains ranked among the 20 most abundant fungal OTUs in the 454-pyrosequencing dataset. This study is one of the most comprehensive comparisons of microbial communities from hydrocarbon-contaminated soils using both isolation and high-throughput sequencing methods.     

Degradation of cellulose by basidiomycetous fungi

Cellulose is the main polymeric component of the plant cell wall, the most abundant polysaccharide on Earth, and an important renewable resource. Basidiomycetous fungi belong to its most potent degraders because many species grow on dead wood or litter, in environment rich in cellulose. Fungal cellulolytic systems differ from the complex cellulolytic systems of bacteria. For the degradation of cellulose, basidiomycetes utilize a set of hydrolytic enzymes typically composed of endoglucanase, cellobiohydrolase and beta-glucosidase. In some species, the absence of cellobiohydrolase is substituted by the production of processive endoglucanases combining the properties of both of these enzymes. In addition, systems producing hydroxyl radicals based on cellobiose dehydrogenase, quinone redox cycling or glycopeptide-based Fenton reaction are involved in the degradation of several plant cell wall components, including cellulose. The complete cellulolytic complex used by a single fungal species is typically composed of more than one of the above mechanisms that contribute to the utilization of cellulose as a source of carbon or energy or degrade it to ensure fast substrate colonization. The efficiency and regulation of cellulose degradation differs among wood-rotting, litter-decomposing, mycorrhizal or plant pathogenic fungi and yeasts due to the different roles of cellulose degradation in the physiology and ecology of the individual groups.