Fungal communities in soils under global change.

Soil fungi play indispensable roles in all ecosystems including the recycling of organic matter and interactions with plants, both as symbionts and pathogens. Past observations and experimental manipulations indicate that projected global change effects, including the increase of CO2 concentration, temperature, change of precipitation and nitrogen (N) deposition, affect fungal species and communities in soils. Although the observed effects depend on the size and duration of change and reflect local conditions, increased N deposition seems to have the most profound effect on fungal communities. The plant-mutualistic fungal guilds - ectomycorrhizal fungi and arbuscular mycorrhizal fungi - appear to be especially responsive to global change factors with N deposition and warming seemingly having the strongest adverse effects. While global change effects on fungal biodiversity seem to be limited, multiple studies demonstrate increases in abundance and dispersal of plant pathogenic fungi. Additionally, ecosystems weakened by global change-induced phenomena, such as drought, are more vulnerable to pathogen outbreaks. The shift from mutualistic fungi to plant pathogens is likely the largest potential threat for the future functioning of natural and managed ecosystems. However, our ability to predict global change effects on fungi is still insufficient and requires further experimental work and long-term observations. Citation: Baldrian P, Bell-Dereske L, Lepinay C, Vetrovský T, Kohout P (2022). Fungal communities in soils under global change. Studies in Mycology 103: 1-24. doi: 10.3114/sim.2022.103.01.

When drought meets forest management: Effects on the soil microbial community of a Holm oak forest ecosystem.

The growth and survival of plants in semiarid Mediterranean forests can be improved through the benefits conferred by thinning, a forest management practice that removes trees and reduces the competition between the remaining ones. Here, we evaluate the impacts of induced drought (the exclusion of 25% of the natural rainfall for 5 years) and thinning, and their interaction, with the objective of determining whether the thinning of Holm oak (Quercus ilex L.) modulates the resistance of the soil microbial community to drought. Sequencing of 16S rRNA and ITS amplicons revealed that drought, thinning, and their interaction influenced the composition of the bacterial community, while the fungal community was exclusively affected by thinning. Thinning consisted of the removal of the aboveground parts of the Holm oak trees, which were thereafter left in forest stand. Thinning contributed to the C and N contents, with parallel increases in microbial biomass, particularly in summer. Drought increased the amounts of total organic C and total N, likely due to the reduced enzyme activities. Indeed, the composition of the bacterial community was modulated primarily by the indirect and long-term effects of drought - the accumulation of soil organic matter - rather than by the direct effect of the lower water content imposed by the drought treatments. Thinning under drought conditions did not increase soil organic C (SOC) content. However, the resistance of the soil microbial community to drought was fostered by thinning, particularly at the functional level, as indicated by the enzyme activities related to C, N and P cycles. These responses were associated to variations in the composition of the microbial communities in thinned, drought-exposed plots, in comparison to unthinned, drought-exposed plots. In conclusion, the interaction between forest management and drought influenced the soil microbial community of a Holm oak-dominated Mediterranean ecosystem.

Chemical and microbiological characterization of an aged PCB-contaminated soil.

This study was aimed at complex characterization of three soil samples (bulk soil, topsoil and rhizosphere soil) from a site historically contaminated with polychlorinated biphenyls (PCB). The bulk soil was the most highly contaminated, with a PCB concentration of 705.95 mg kg(-1), while the rhizosphere soil was the least contaminated (169.36 mg kg(-1)). PCB degradation intermediates, namely chlorobenzoic acids (CBAs), were detected in all the soil samples, suggesting the occurrence of microbial transformation processes over time. The higher content of organic carbon in the topsoil and rhizosphere soil than in the bulk soil could be linked to the reduced bioaccessibility (bioavailability) of these chlorinated pollutants. However, different proportions of the PCB congener contents and different bioaccessibility of the PCB homologues indicate microbial biotransformation of the compounds. The higher content of organic carbon probably also promoted the growth of microorganisms, as revealed by phospholipid fatty acid (PFLA) quantification. Tag-encoded pyrosequencing analysis showed that the bacterial community structure was significantly similar among the three soils and was predominated by Proteobacteria (44-48%) in all cases. Moreover, analysis at lower taxonomic levels pointed to the presence of genera (Sphingomonas, Bulkholderia, Arthrobacter, Bacillus) including members with reported PCB removal abilities. The fungal community was mostly represented by Basidiomycota and Ascomycota, which accounted for >80% of all the sequences detected in the three soils. Fungal taxa with biodegradation potential (Paxillus, Cryptococcus, Phoma, Mortierella) were also found. These results highlight the potential of the indigenous consortia present at the site as a starting point for PCB bioremediation processes.

Local distribution of ectomycorrhizae-associated basidiomycetes in forest soil correlates with the degree of soil organic matter humification and available electrolytes.

Spatial distribution of ectomycorrhizae-associated basidiomycetes was determined in oakbirch forest using terminal restriction fragment length polymorphism (T-RFLP) analysis. The data were correlated with actual soil humidity, pH, electric conductivity of the soil extract, absorbance A(465) and A(665) of water and alkali soil extracts and with the ratio A(465)/A(665) (parameter A4/A6). Natural non-homogeneity of the soil parameters was used as experimental gradient. Distance-based redundancy analysis of the T-RFLP data (with soil parameters being taken as environmental parameters) provided significant results when ITS1F-terminanted restriction fragments were analyzed. Among other fungi, a Mycena galericulata related fungus was observed to correlate negatively with A4/A6, indicating its association with highly humified soil organic matter. Positive association of other, unidentified fungi with A4/A6 was also observed. Several other unidentified fungi negatively correlated with electric conductivity of the soil extract. The results may explain nonhomogeneity of the spatial distribution of the fungi associated with ectomycorrhizae as a result of their interaction with non-homogeneous soil environment.

Production and regulation of lignocellulose-degrading enzymes of Poria-like wood-inhabiting basidiomycetes.

The wood-decomposing fungal species Antrodia macra, A. pulvinascens, Ceriporiopsis aneirina, C. resinascens and Dichomitus albidofuscus were determined for production of laccase (LAC), Mn peroxidase (MnP), lignin peroxidase (LiP), endo-l,4-P-beta-glucanase, endo-l,4-beta-xylanase, cellobiohydrolase, 1,4-beta-glucosidase and 1,4-beta-xylosidase. The results confirmed the brown-rot mode of Antrodia spp. which did not produce the activity of LAC and MnP. The remaining species performed detectable activity of both enzymes while no strain produced LiP. Significant inhibition of LAC production by high nitrogen was found in all white-rot species while only MnP of D. albidofuscus was regulated in the same way. The endoglucanase and endoxylanase activities of white-rotting species were inhibited by glucose in the medium while those of Antrodia spp. were not influenced by glucose concentration. The regulation of enzyme activity and bio-mass production can vary even within a single fungal genus.

Influence of mineral and organic fertilization on soil fungi, enzyme activities and humic substances in a long-term field experiment.

Changes in microfungal communities, fungal activities and humic substances (HS) in agricultural soils kept under different fertilization regimes were observed and their causal relationships were investigated in a long-term field experiment. Fertilization did not change the abundance of HS-utilizing microfungi and, except for organic amendment alone, total culturable microfungi were also unaffected by this factor. Organic fertilization increased activities of manganese peroxidase (MnP) and proteinase, but decreased endo-1,4-beta-glucanase activity compared to the corresponding control without organic fertilization. In soils treated with mineral fertilizers, the activities of MnP, endo-1,4-beta-glucanase and proteinase were higher than in control without any mineral treatment. Both the aromaticity of fulvic acid and the molar mass of humic acid was lower in soil with organic fertilization, which may be a result of oxidative degradation mediated by higher MnP activity observed in treatments with organic fertilization.

Temperature affects the production, activity and stability of ligninolytic enzymes in Pleurotus ostreatus and Trametes versicolor.

Enzyme activity was determined in cultures of Pleurotus ostreatus and Trametes versicolor with cellulose as a sole C source and high C/N ratio. The fungi were able to grow and produce laccase and Mn-peroxidase (MnP) at 5-35 degrees C, the highest production being recorded at 25-30 degrees C in P. ostreatus and at 35 degrees C in T. versicolor. Production of both enzymes at 10 degrees C accounted only for 4-20% of the maximum value. Temperature optima for enzyme activity were 50 and 55 degrees C for P. ostreatus and T. versicolor laccases, respectively, and 60 degrees C for MnP. Temperatures causing 50% loss of activity after 24 h were 32 and 47 degrees C for laccases and 36 and 30 degrees C for MnP from P. ostreatus and T. versicolor, respectively.

The use of hydroxyl-radical-generating systems for the treatment of olive mill wastewaters.

Three hydroxyl-radical producing biomimetic systems, composed of CuII, hydrogen peroxide and pyridine, glucaric or succinic acid, were able to perform decolorization of olive mill wastewaters (OMW) >85 % within 3 d combined with a significant removal of total phenols (>75 %). The systems consisting of 50 mmol/L succinic acid, 5-10 mmol/L CuSO4 and 100 mmol/L H2O2 were the most effective at OMW treatment, and led to the reduction of phenol contents to <1 % along with high decolorization (>88 %) and acceptable values of chemical oxygen demand.

Production of lignocellulose-degrading enzymes and changes in soil bacterial communities during the growth of Pleurotus ostreatus in soil with different carbon content.

The extracellular enzyme activity and changes in soil bacterial community during the growth of the ligninolytic fungus Pleurotus ostreatus were determined in nonsterile soil with low and high available carbon content. In soil with P. ostreatus, the activity of ligninolytic enzymes laccase and Mn-peroxidase was several orders of magnitude higher than in soil without the fungus. Addition of lignocellulose to soil increased the activity of cellulolytic fungi and the production of Mn-peroxidase by P. ostreatus. The counts of heterotrophic bacteria were more significantly affected by the presence of lignocellulose than by P. ostreatus. The effects of both substrate addition and time (succession) were more significant factors affecting the soil bacterial community than the presence of P. ostreatus. Bacterial community structure was affected by fungal colonization in low carbon soil, where a decrease of diversity and changes in substrate utilization profiles were detected.

Purification and characterization of laccase from the white-rot fungus Daedalea quercina and decolorization of synthetic dyes by the enzyme.

The white-rot fungus Daedalea quercina produced the ligninolytic enzymes laccase and Mn-dependent peroxidase. Laccase was purified using anionexchange and size-exclusion chromatographies. SDS-PAGE showed the purified laccase to be a monomeric protein of 69 kDa (71 kDa using gel filtration) with an isoelectric point near 3.0. The optimum pH for activity was below 2.0 for 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (K(m)=38 microM), 4.0 for 2,6-dimethoxyphenol (K(m)=48 microM), 4.5 for guaiacol (K(m)=93 microM) and 7.0 for syringaldazine (K(m)=131 microM). The temperature optimum was between 60 and 70 degrees C depending on the pH and buffer used. The enzyme was stable up to 45 degrees C, and stability was higher at alkaline pH. Enzyme activity was increased by the addition of Cu(2+) and inhibited by Mn(2+), sodium azide, dithiothreitol, and cysteine. Laccase from Daedalea quercina was able to decolorize the synthetic dyes Chicago sky blue, poly B-411, remazol brilliant blue R, trypan blue and reactive blue 2.

Variability of laccase activity in the white-rot basidiomycete Pleurotus ostreatus.

The production of laccase in liquid cultures of the white-rot fungus Pleurotus ostreatus was highly variable. During the first days of cultivation, the relative variability was as high as 80-100% and it decreased to 30% in the course of cultivation. The main source of variability was assumed to be the independent development of enzyme activity in individual cultures. Cultures with high laccase production showed also high production of the other ligninolytic enzyme--Mn-dependent peroxidase. The variability was probably due to the source of inoculum, deactivation of the enzyme in culture liquid and genetic variations among the cultures. Variability of laccase activities was lower during solid-state fermentation on wheat straw and during the growth in nonsterile soil.

Decolorization of synthetic dyes by the Fenton reagent and the Cu/pyridine/H2O2 system.

Representative azo, triphenylmethane, heterocyclic and polymeric synthetic dyes have been decolorized by two biological non-ezymatic systems, copper/pyridine/H2O2 and the Fenton reagent. With the former system, intensive decolorization measured after 1 h was obtained with phenol red (89%), tropaeolin 00 (58%), Evans blue (95%), eosin yellowish (84%), and Poly B-411 (92%). The rate of decolorization was not affected by pH in the range of 3-9 and increased with increasing temperature. The use of the radical scavengers thiourea and superoxide dismutase showed that hydroxyl radicals rather than superoxide anions are involved in the reaction. Omission of pyridine led to a substantial decrease in the extent of decolorization (20-50% decolorization). The use of organic peroxide instead of H2O2 resulted in slightly slower decolorization, similar values of decolorization being obtained only after a 2-h incubation. Decolorization of the dyes by the Fenton reagent was also very effective but slower than that obtained with the first system. Except for phenol red and eosin yellowish, (decolorization 8% and 52%, respectively) the dyes were decolorized up to 99% after 1-day incubation.

Copper sorption by native and modified pellets of wood-rotting basidiomycetes.

AIMS: The aim of this study was to investigate the biosorption of copper to the pellets of different wood-rotting fungal species. METHODS AND RESULTS: Copper sorption was studied in both batch and column arrangements. The optimum pH for copper sorption was between 3.5 and 4. In 100 mg l(-1) Cu (II), maximum qe values were found for Oudemansiella mucida (8.77 mg g(-1) dry wt), Lepista nuda (6.29 mg g(-1)), Pycnoporus cinnabarinus (5.08 mg g(-1)) and Pleurotus ostreatus (4.77 mg g(-1)). Both biomass yield and specific sorption were influenced by the composition of the fermentation broth. The results of column experiments showed that mycelial pellets of wood-rotting fungi can be considered as promising biosorbent material. CONCLUSIONS: Pellets of wood-rotting fungi showed the same or better copper sorption properties as those previously reported for lower fungi or filamentous bacteria, as well as good mechanical properties.

Influence of cadmium and mercury on activities of ligninolytic enzymes and degradation of polycyclic aromatic hydrocarbons by Pleurotus ostreatus in soil.

The white-rot fungus Pleurotus ostreatus was able to degrade the polycyclic aromatic hydrocarbons (PAHs) benzo[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, and benzo[ghi]perylene in nonsterile soil both in the presence and in the absence of cadmium and mercury. During 15 weeks of incubation, recovery of individual compounds was 16 to 69% in soil without additional metal. While soil microflora contributed mostly to degradation of pyrene (82%) and benzo[a]anthracene (41%), the fungus enhanced the disappearance of less-soluble polycyclic aromatic compounds containing five or six aromatic rings. Although the heavy metals in the soil affected the activity of ligninolytic enzymes produced by the fungus (laccase and Mn-dependent peroxidase), no decrease in PAH degradation was found in soil containing Cd or Hg at 10 to 100 ppm. In the presence of cadmium at 500 ppm in soil, degradation of PAHs by soil microflora was not affected whereas the contribution of fungus was negligible, probably due to the absence of Mn-dependent peroxidase activity. In the presence of Hg at 50 to 100 ppm or Cd at 100 to 500 ppm, the extent of soil colonization by the fungus was limited.

Screening of Pleurotus ostreatus isolates for their ligninolytic properties during cultivation on natural substrates.

Thirteen basidiospore-derived isolates of Pleurotus ostreatus f6 strain differing in the level of ligninolytic enzyme production and other characteristics (mycelium extension rate, colony morphology) from the parental strain were cultivated on natural substrates. Under these conditions ligninolytic enzyme activity, loss of organic mass, polycyclic aromatic hydrocarbons (PAHs) degradation and colonization of sterile and nonsterile soil were studied. The activity of ligninolytic enzymes was substantially higher in straw than in liquid culture, although the differences between the isolates were less pronounced on this substrate. Some of the isolates showed a very good ability to decompose the lignocellulosic substrate (straw) and a relatively high loss of organic mass was found after 50 days of cultivation in these strains. The original strain f6 and isolates B13 and B26 successfully degraded all seven tested PAH compounds present in experimental soil samples, but the higher or lower ligninolytic enzyme production of isolates tested had no substantial effect on the extent of the degradation. In our screening, six basidiospore-derived isolates growing well in nonsterile soil were found, which could be suitable for the prospective biotechnological exploitation.

Degradation of polycyclic aromatic hydrocarbons by the copper(II)-hydrogen peroxide system.

A non-enzymic system containing CuSO4 (10 mmol/L) and hydrogen peroxide (100 mmol/L) was used for the degradation of three polycyclic aromatic hydrocarbons: phenanthrene, fluoranthene, and pyrene (all at 10 mmol/L). The system degraded the compounds rapidly and efficiently. After 1 d at room temperature, more than 80% of pyrene, phenanthrene, and fluoranthene disappeared. Several products are formed during the reaction including a black precipitate.