Recent advances in exploring physiology and biodiversity of ectomycorrhizas highlight the functioning of these symbioses in ecosystems

Ectomycorrhizas, the dominating mycorrhizal symbiosis in boreal, temperate and some tropical forests, are formed by 5000-6000 species of the asco- and basidiomycetes. This high diversity of fungal partners allows optimal foraging and mobilisation of various nitrogen and phosphorus forms from organic soil layers. In this review, two approaches to study the functioning of this multitude of symbiotic associations are presented. On selected culture models, physiological and molecular investigations have shown that the supply of hexoses has a key function in controlling the plant^fungus interaction via partner-specific regulation of gene expression. Environmental factors which affect fungal carbon supply, such as increased nitrogen availability, also affect mycorrhiza formation. Based on such laboratory results, the adaptative capability of ectomycorrhizas to changing field conditions is discussed. The second approach consists of analysing the distribution of mycorrhizas in ecosystem compartments and to relate distribution patterns to variations of ecological factors. Recent advances in identification of fungal partners in ectomycorrhizas by analysing the internal transcribed spacer of ribosomal DNA are presented, which can help to resolve sampling problems in field studies. The limits of the laboratory and the field approaches are discussed. Despite some problems, this combined approach is the most promising. Direct investigation of gene expression, which has been introduced for soil bacteria, will be difficult in the case of mycorrhizal fungi which constitute organisms with functionally varying structure

Rotating disk electrodes to assess river biofilm thickness and elasticity

The present study examined the relevance of an electrochemical method based on a rotating disk electrode (RDE) to assess river biofilm thickness and elasticity. An in situ colonisation experiment in the River Garonne (France) in August 2009 sought to obtain natural river biofilms exhibiting differentiated architecture. A constricted pipe providing two contrasted flow conditions (about 0.1 and 0.45 m s−1 in inflow and constricted sections respectively) and containing 24 RDE was immersed in the river for 21 days. Biofilm thickness and elasticity were quantified using an electrochemical assay on 7 and 21 days old RDE-grown biofilms (t7 and t21, respectively). Biofilm thickness was affected by colonisation length and flow conditions and ranged from 36 ± 15 μm (mean ± standard deviation, n = 6) in the fast flow section at t7 to 340 ± 140 μm (n = 3) in the slow flow section at t21. Comparing the electrochemical signal to stereomicroscopic estimates of biofilms thickness indicated that the method consistently allowed (i) to detect early biofilm colonisation in the river and (ii) to measure biofilm thickness of up to a few hundred μm. Biofilm elasticity, i.e. biofilm squeeze by hydrodynamic constraint, was significantly higher in the slow (1300 ± 480 μm rpm1/2, n = 8) than in the fast flow sections (790 ± 350 μm rpm1/2, n = 11). Diatom and bacterial density, and biofilm-covered RDE surface analyses (i) confirmed that microbial accrual resulted in biofilm formation on the RDE surface, and (ii) indicated that thickness and elasticity represent useful integrative parameters of biofilm architecture that could be measured on natural river assemblages using the proposed electrochemical method

Microbial dynamics associated with leaves decomposing in the mainstem and floodplain pond of a large river

Aquatic habitats of forested floodplain systems receive large inputs of allochthonous plant litter. We examined the decomposition of, and microbial productivity associated with, leaves of a common floodplain tree, Populus gr. nigra, in the mainstem and floodplain pond of a seventh order river in 2 consecutive years. Litter bags were submerged at both sites, retrieved periodically, and analyzed for litter mass loss, bacterial and fungal biomass, growth rate and production, and sporulation rates of aquatic hyphomycetes. Litter decomposition rates were similar in both sites and years (leaf breakdown coefficients k of 0.0070 to 0.0085 d–1), although microbial dynamics partly differed between sites. Species diversity of aquatic hyphomycetes was lower on leaves submerged in the pond (16 species) than in the river (21 species). Mycelial biomass was also significantly lower in the pond, with values <20 mgCg–1 of detrital C, whereas peaks of 50 and 80 mgC g–1 were reached in leaves in the mainstem. These differences contrast with the comparable fungal productivity at both sites (peak rates of 1.4 mg of mycelial C per g of detrital C per day in both years). This suggests that fungi were equally productive in both habitats but experienced greater losses in the pond. Bacterial numbers and biomass also showed the same basic pattern at both sites, although somewhat higher levels were reached in the pond (maximum of about 10^10 cells and 0.5 mg g–1 of detrital C). Bacterial- specific production rates fluctuated between 0.06 and 1.5 d–1 with lower values occurring in the floodplain pond. Although bacteria on leaves were clearly outweighed by fungi in terms of biomass, they accounted for a sizeable fraction of the total biomass (up to 11%), and up to 32% of the total microbial production. Our comparison of bacterial and fungal productivity thus points to a critical role of fungi in litter decomposition in aquatic habitats of river floodplain systems, while suggesting that bacteria must not be overlooked as important agents of litter decompositon in riverine environments

Microbial decomposers communities are mainly structured by trophic status in circumneutral and alkaline streams

In streams, the release of nitrogen and phosphorus is reported to affect microbial communities and the ecological processes they govern. Moreover, the type of inorganic nitrogen (NO3, NO2 or NH4) may differently impact microbial communities. We aimed to identify the environmental factors that structure aquatic microbial communities and drive leaf-litter decomposition along a gradient of eutrophication. We selected 5 circumneutral (Portuguese) and 5 alkaline (French) streams differing in nutrient concentrations to monitor mass loss of alder leaves, bacterial and fungal diversity by PCR-DGGE, fungal biomass and reproduction, and bacterial biomass during 11 weeks of leaf immersion. The concentrations of inorganic nutrients in the stream water ranged from 5 - 300 µg L-1 SRP; 0.30 – 5.50 mg L-1 NO3-N; 2 - 103 µg L-1 NO2-N; and < 4 - 7,100 µg L-1 NH4-N. Species richness was maximum in moderately anthropized (eutrophic) streams but decreased in the most anthropized (hypertrophic) streams. Different species assemblages were found in subsets of streams of different trophic status. In both geographic areas, the limiting nutrient, either nitrate or phosphate, stimulated the microbial activity in streams of intermediate trophic status. In the hypertrophic streams, fungal biomass and reproduction were significantly lower and bacterial biomass dramatically decreased at the site with the highest ammonium concentration. The limiting nutrients that defined the trophic status were the main factor structuring fungal and bacterial communities whatever the geographic area. Very high ammonium concentration in the stream water most probably has negative impacts on microbial decomposer communities

Electroactivity of phototrophic river biofilms and constitutive cultivable bacteria

Electroactivity is a property of microorganisms assembled in biofilms that has been highlighted in a variety of environments. This characteristic was assessed for phototrophic river biofilms at the community scale and at the bacterial population scale. At the community scale, electroactivity was evaluated on stainless steel and copper alloy coupons used both as biofilm colonization supports and as working electrodes. At the population scale, the ability of environmental bacterial strains to catalyze oxygen reduction was assessed by cyclic voltammetry. Our data demonstrate that phototrophic river biofilm development on the electrodes, measured by dry mass and chlorophyll a content, resulted in significant increases of the recorded potentials, with potentials of up to +120 mV/saturated calomel electrode (SCE) on stainless steel electrodes and +60 mV/SCE on copper electrodes. Thirty-two bacterial strains isolated from natural phototrophic river biofilms were tested by cyclic voltammetry. Twenty-five were able to catalyze oxygen reduction, with shifts of potential ranging from 0.06 to 0.23 V, cathodic peak potentials ranging from −0.36 to −0.76 V/SCE, and peak amplitudes ranging from −9.5 to −19.4 μA. These isolates were diversified phylogenetically (Actinobacteria, Firmicutes, Bacteroidetes, and Alpha-, Beta-, and Gammaproteobacteria) and exhibited various phenotypic properties (Gram stain, oxidase, and catalase characteristics). These data suggest that phototrophic river biofilm communities and/or most of their constitutive bacterial populations present the ability to promote electronic exchange with a metallic electrode, supporting the following possibilities: (i) development of electrochemistry-based sensors allowing in situ phototrophic river biofilm detection and (ii) production of microbial fuel cell inocula under oligotrophic conditions

Non-target effects of three formulated pesticides on microbially-mediated processes in a clay-loam soil

An experiment was performed to study non-target effects of difenoconazole (fungicide), deltamethrin (insecticide) and ethofumesate (herbicide) on microbial parameters in a clay-loam soil. Pesticides were applied as commercial formulations to soil samples at different concentrations (5, 50 and 500 mg kg−1 DW soil) and then incubated under laboratory conditions for 3 months. Throughout the incubation period, microbial parameters were determined at days 7, 30, 60 and 90. At 5 mg kg−1 DW soil, none of the three pesticides caused significant changes in soil microbial parameters. In contrast, at 500 mg kg−1 DW soil, pesticide application decreased overall soil microbial activity, negatively affecting the activity of soil enzymes. Similarly, at 500 mg kg−1 DW soil, difenoconazole and ethofumesate, but not deltamethrin, caused a pesticide-induced stress on soil microbial communities, as reflected by the respiratory quotient. Besides, deltamethrin and ethofumesate at 50 and 500 mg kg−1 DW soil resulted in lower values of denitrification potential. It was concluded that, although pesticide concentration had a somewhat inconsistent and erratic effect on soil microbial parameters, pesticide application at 500 mg kg−1 DW soil did have an impact on many of the microbial parameters studied here

Effects of herbicide mixtures on freshwater microalgae with the potential effect of a safener

International audienceFreshwater microalgae are primary producers and cosmopolitan species subjected to the effects of herbicides. In this work, the in vitro algal growth inhibitory effects of 11 agrochemicals (9 herbicides, 1 metabolite, and 1 safener) were quantified. Chemical compounds were applied singly and in specific mixtures. Three species were used in axenic condition: the green alga Desmodesmus subspicatus (Chodat), the diatoms Nitzschia palea (Kützing) W. Smith and Navicula pelliculosa (Kützing) Hilse. When exposed to single compounds, N. palea and N. pelliculosa were only sensitive to atrazine/desethylatrazine and the safener benoxacor (BE), respectively. D. subspicatus was equally sensitive to four herbicides including atrazine and its metabolite and significantly more sensitive to iodosulfuron-methyl-sodium (IODO). The mixture of these five compounds induced a significantly higher growth inhibition of about 1.5-fold than IODO alone, which could be attributed to the four other herbicides. The mixture of all compounds was twofold less toxic than IODO on D. subspicatus. A halogen atom is present in IODO as in the herbicides to which the safener BE À known to induce glutathione-S-transferases À is associated in agrochemical preparations. We then showed that IODO was less toxic when combined with non-toxic concentrations of BE. These results indicated that the toxicity of the most active herbicide studied was decreased by a non-herbicide compound present in agrochemical formulations of other herbicides. These results suggest the importance to take into account the chemistry and the mechanisms of action for each compound in a risk assessment approach of a complex mixture

Light interception principally drives the understory response to boxelder invasion in riparian forests

Since several decades, American boxelder (Acer negundo) is replacing white willow (Salix alba) riparian forests along southern European rivers. This study aims to evaluate the consequences of boxelder invasion on understory community in riparian areas. We determined the understory species richness, composition and biomass in boxelder and white willow stands located in three riparian forests, representative of three rivers with distinct hydrological regimes. We investigated correlation of these variables to soil moisture and particle size, main soil nutrient stocks, potential nitrification and denitrification, tree canopy cover and photosynthetic active radiation (PAR) at the ground level. A greenhouse experiment was then conducted to identify the causal factors responsible for changes in the understory. The effect of soil type, PAR level and water level on the growth and the biomass production of Urtica dioica were examined. A lower plant species richness and biomass, and a modification of community composition were observed for boxelder understory in all sites, regardless of their environmental characteristics. The strongest modification that follows boxelder invasion was the decline in U. dioica, the dominant species of the white willow forest understory. These differences were mainly correlated with a lower incident PAR under boxelder canopy. The greenhouse experiment identified PAR level as the main factor responsible for the changes in U. dioica stem number and biomass. Our results indicate that adult boxelder acts as an ecosystem engineer that decreases light availability. The opportunistic invasion by boxelder leads to important understory changes, which could alter riparian ecosystem functioning

Effect of Culture Conditions on Ergosterol as an Indicator of Biomass in the Aquatic Hyphomycetes

Ergosterol is a membrane component specific to fungi that can be used to estimate fungal biomass using appropriate factors of conversion. Our objectives were to determine the limits of use of ergosterol content as a measure of biomass for aquatic hyphomycetes, and to evaluate a previously established ergosterol-to-biomass conversion factor. We varied inoculum quality, growth medium, and degree of shaking of four aquatic hyphomycete species. In cultures inoculated with homogenized mycelium, we found a significant effect of shaking condition and culture age on ergosterol content. In liquid cultures with defined medium, ergosterol content reached 10 to 11 μg/mg of mycelium (dry mass) and varied by factors of 2.2 during exponential growth and 1.3 during stationary phase. The increase in ergosterol content during exponential phase could be attributed, at least in part, to rapid depletion of glucose. Oxygen availability to internal hyphae within the mycelial mass is also responsible for the differences found between culture conditions. Ergosterol concentration ranged from 0.8 to 1.6 μg/mg in static cultures inoculated with agar plugs. Ergosterol content varied by a factor of 4 in two media of different richnesses. For different combinations of these parameters, strong (r(2) = 0.83 to 0.98) and highly significant (P ≪ 0.001) linear relationships between ergosterol and mycelial dry mass (up to 110 mg) were observed. Overall, the ergosterol content varied by a factor of 14 (0.8 to 11 mg/g). These results suggest that care must be taken when the ergosterol content is used to compare data generated in different field environments