Monique Carnol

Biotic homogenization can decrease landscape-scale forest multifunctionality

Significance Numerous studies have demonstrated the importance of biodiversity in maintaining multiple ecosystem functions and services (multifunctionality) at local spatial scales, but it is unknown whether similar relationships are found at larger spatial scales in real-world landscapes. Here, we show, for the first time to our knowledge, that biodiversity can also be important for multifunctionality at larger spatial scales in European forest landscapes. Both high local (α-) diversity and a high turnover in species composition between locations (high β-diversity) were found to be potentially important drivers of ecosystem multifunctionality. Our study provides evidence that it is important to conserve the landscape-scale biodiversity that is being eroded by biotic homogenization if ecosystem multifunctionality is to be maintained. Many experiments have shown that local biodiversity loss impairs the ability of ecosystems to maintain multiple ecosystem functions at high levels (multifunctionality). In contrast, the role of biodiversity in driving ecosystem multifunctionality at landscape scales remains unresolved. We used a comprehensive pan-European dataset, including 16 ecosystem functions measured in 209 forest plots across six European countries, and performed simulations to investigate how local plot-scale richness of tree species (α-diversity) and their turnover between plots (β-diversity) are related to landscape-scale multifunctionality. After accounting for variation in environmental conditions, we found that relationships between α-diversity and landscape-scale multifunctionality varied from positive to negative depending on the multifunctionality metric used. In contrast, when significant, relationships between β-diversity and landscape-scale multifunctionality were always positive, because a high spatial turnover in species composition was closely related to a high spatial turnover in functions that were supported at high levels. Our findings have major implications for forest management and indicate that biotic homogenization can have previously unrecognized and negative consequences for large-scale ecosystem multifunctionality.

Nitrosomonas europaea-like bacteria detected as the dominant β-subclass Proteobacteria ammonia oxidisers in reference and limed acid forest soils

Net nitrification in intact soil cores and the community of ammonia-oxidising bacteria were studied in acid Norway spruce (Picea abies (L.) Karst) and sessile oak (Quercus petraea (Matt. Lieb.)) soils (Haute Ardenne, east Belgium) 18 months after treatment with 5tha1 dolomite lime. Liming caused a significant increase in nitrification in the upper soil layers (0.15m) of both stands. DGGE (denaturing gradient gel electrophoresis) profiling after -subclass ammonia oxidiser-specific polymerase chain reaction (PCR), combined with hybridisation and sequencing of excised DGGE bands revealed a dominance of Nitrosomonas europaea-like sequences, independent of soil horizon, tree species and lime treatment. A minority Nitrosospira-like population was detected, which showed affinity to nitrosospiras previously detected in acid soil. These results contrast with several reports suggesting a dominance of Nitrosospira-like organisms among ammonia oxidiser communities in acid soils. [KEYWORDS: 16S rDNA; DGGE; Picea abies; Quercus petraea; Nitrification; Dolomite lime]

The effects of ammonium sulphate deposition and root sinks on soil solution chemistry in coniferous forest soils

The effects of enhanced (NH42SO4deposition on soil solution cation and anion concentrations and annualionic fluxes were followed using a standardised experimental protocolin six European coniferous forests with contrasting soil types, pollutioninputs and climate. Native soil cores containing a ceramic suction cupwere installed in the field, roofed and watered every two weeks withlocal throughfall or local throughfall with added(NH4)2SO4 at 75 kgNH4+-N ha-1 a-1. Livingroot systems were established in half of the lysimeters.Untreated throughfall NH4+-N deposition at thesites ranged from 3.7 to 29 kg ha-1 a-1Soil leachates were collected at two weekly intervalsover 12 months and analysed for volume, andconcentrations of major anions and cations. Increasesin soil solution NO3- concentrations inresponse to N additions were observed after 4–9months at three sites, whilst one sandy soil with highC:N ratio failed to nitrify under any of thetreatments. Changes in NO3- concentrationsin soil solution controlled soil solution cationconcentrations in the five nitrifying soils, withAl3+ being the dominant cation in the more acidsoils with low base saturation. The acidification responses ofthe soils to the (NH42SO4additions were primarily related to the ability of thesoils to nitrify the added NH4+. pH and soiltexture seemed important in controllingNH4+ leaching in response to the treatments,with two less acidic, clay/clay loam sites showingalmost total retention of added NH4+, whilstnearly 75% of the added N was leached asNH4+ at the acid sandy soils. The presenceof living roots significantly reduced soil solutionNO3- and associated cation concentrations attwo of the six sites. The very different responses of the sixsoils to increased (NH4)2SO4deposition emphasise that the establishment of N critical loadsfor forest soils need to allow for differences in N storagecapacity and nitrification potential.

Biodiversity and ecosystem functioning relations in European forests depend on environmental context

The importance of biodiversity in supporting ecosystem functioning is generally well accepted. However, most evidence comes from small-scale studies, and scaling-up patterns of biodiversity-ecosystem functioning (B-EF) remains challenging, in part because the importance of environmental factors in shaping B-EF relations is poorly understood. Using a forest research platform in which 26 ecosystem functions were measured along gradients of tree species richness in six regions across Europe, we investigated the extent and the potential drivers of context dependency of B-EF relations. Despite considerable variation in species richness effects across the continent, we found a tendency for stronger B-EF relations in drier climates as well as in areas with longer growing seasons and more functionally diverse tree species. The importance of water availability in driving context dependency suggests that as water limitation increases under climate change, biodiversity may become even more important to support high levels of functioning in European forests.

A Phenotypic and Genotypic Analysis of the Antimicrobial Potential of Cultivable Streptomyces Isolated from Cave Moonmilk Deposits

Moonmilk speleothems of limestone caves host a rich microbiome, among which Actinobacteria represent one of the most abundant phyla. Ancient medical texts reported that moonmilk had therapeutical properties, thereby suggesting that its filamentous endemic actinobacterial population might be a source of natural products useful in human treatment. In this work, a screening approach was undertaken in order to isolate cultivable Actinobacteria from moonmilk of the Grotte des Collemboles in Belgium, to evaluate their taxonomic profile, and to assess their potential in biosynthesis of antimicrobials. Phylogenetic analysis revealed that all 78 isolates were exclusively affiliated to the genus Streptomyces and clustered into 31 distinct phylotypes displaying various pigmentation patterns and morphological features. Phylotype representatives were tested for antibacterial and antifungal activities and their genomes were mined for secondary metabolite biosynthetic genes coding for non-ribosomal peptide synthetases (NRPSs), and polyketide synthases (PKS). The moonmilk Streptomyces collection was found to display strong inhibitory activities against a wide range of reference organisms, as 94, 71, and 94% of the isolates inhibited or impaired the growth of Gram-positive, Gram-negative bacteria, and fungi, respectively. Interestingly, 90% of the cave strains induced strong growth suppression against the multi-drug resistant Rasamsonia argillacea, a causative agent of invasive mycosis in cystic fibrosis and chronic granulomatous diseases. No correlation was observed between the global antimicrobial activity of an individual strain and the number of NRPS and PKS genes predicted in its genome, suggesting that approaches for awakening cryptic metabolites biosynthesis should be applied to isolates with no antimicrobial phenotype. Overall, our work supports the common belief that moonmilk might effectively treat various infectious diseases thanks to the presence of a highly diverse population of prolific antimicrobial producing Streptomyces, and thus may indeed constitute a promising reservoir of potentially novel active natural compounds.

Functional analysis of the three HMA4 copies of the metal hyperaccumulator Arabidopsis halleri

Highlight Expression of the Arabidopsis halleri hyperaccumulation gene HMA4 in A. thaliana reveals functional differentiation among the three AhHMA4 copies and non-polar localization of the AhHMA4 protein in root pericycle cells.

Zinc triggers a complex transcriptional and post-transcriptional regulation of the metal homeostasis gene FRD3 in Arabidopsis relatives

Highlight High expression of the FRD3 gene in the metal hyperaccumulator Arabidopsis halleri stems from complex zinc-regulated transcriptional and post-transcriptional mechanisms present in the non-hyperaccumulating Arabidopsis thaliana.

Assessment of the Potential Role of Streptomyces in Cave Moonmilk Formation

Moonmilk is a karstic speleothem mainly composed of fine calcium carbonate crystals (CaCO3) with different textures ranging from pasty to hard, in which the contribution of biotic rock-building processes is presumed to involve indigenous microorganisms. The real microbial input in the genesis of moonmilk is difficult to assess leading to controversial hypotheses explaining the origins and the mechanisms (biotic vs. abiotic) involved. In this work, we undertook a comprehensive approach in order to assess the potential role of filamentous bacteria, particularly a collection of moonmilk-originating Streptomyces, in the genesis of this speleothem. Scanning electron microscopy (SEM) confirmed that indigenous filamentous bacteria could indeed participate in moonmilk development by serving as nucleation sites for CaCO3 deposition. The metabolic activities involved in CaCO3 transformation were furthermore assessed in vitro among the collection of moonmilk Streptomyces, which revealed that peptides/amino acids ammonification, and to a lesser extend ureolysis, could be privileged metabolic pathways participating in carbonate precipitation by increasing the pH of the bacterial environment. Additionally, in silico search for the genes involved in biomineralization processes including ureolysis, dissimilatory nitrate reduction to ammonia, active calcium ion transport, and reversible hydration of CO2 allowed to identify genetic predispositions for carbonate precipitation in Streptomyces. Finally, their biomineralization abilities were confirmed by environmental SEM, which allowed to visualize the formation of abundant mineral deposits under laboratory conditions. Overall, our study provides novel evidences that filamentous Actinobacteria could be key protagonists in the genesis of moonmilk through a wide spectrum of biomineralization processes.

Metal binding to the N-terminal cytoplasmic domain of the PIB ATPase HMA4 is required for metal transport in Arabidopsis.

PIB ATPases are metal cation pumps that transport metals across membranes. These proteins possess N- and C-terminal cytoplasmic extensions that contain Cys- and His-rich high affinity metal binding domains, which may be involved in metal sensing, metal ion selectivity and/or in regulation of the pump activity. The PIB ATPase HMA4 (Heavy Metal ATPase 4) plays a central role in metal homeostasis in Arabidopsis thaliana and has a key function in zinc and cadmium hypertolerance and hyperaccumulation in the extremophile plant species Arabidopsis halleri. Here, we examined the function and structure of the N-terminal cytoplasmic metal-binding domain of HMA4. We mutagenized a conserved CCTSE metal-binding motif in the domain and assessed the impact of the mutations on protein function and localization in planta, on metal-binding properties in vitro and on protein structure by Nuclear Magnetic Resonance spectroscopy. The two Cys residues of the motif are essential for the function, but not for localization, of HMA4 in planta, whereas the Glu residue is important but not essential. These residues also determine zinc coordination and affinity. Zinc binding to the N-terminal domain is thus crucial for HMA4 protein function, whereas it is not required to maintain the protein structure. Altogether, combining in vivo and in vitro approaches in our study provides insights towards the molecular understanding of metal transport and specificity of metal P-type ATPases.

AOB community structure and richness under European beech, sessile oak, Norway spruce and Douglas-fir at three temperate forest sites

Background and aimsThe relations between tree species, microbial diversity and activity can alter ecosystem functioning. We investigated ammonia oxidizing bacteria (AOB) community structure and richness, microbial/environmental factors related to AOB diversity and the relationship between AOB diversity and the nitrification process under several tree species.MethodsForest floor (Of, Oh) was sampled under European beech, sessile oak, Norway spruce and Douglas-fir at three sites. AOB community structure was assessed by PCR-DGGE and sequencing. Samples were analyzed for net N mineralization, potential nitrification, basal respiration, microbial biomass, microbial or metabolic quotient, pH, total nitrogen, extractable ammonium, organic matter content and exchangeable cations.ResultsAOB community structure and tree species effect on AOB diversity were site-specific. AOB richness was not related to nitrification. Factors regulating ammonium availability, i.e. net N mineralization or microbial biomass, were related to AOB community structure.ConclusionOur research shows that, at larger spatial scales, site specific characteristics may be more important than the nature of tree species in determining AOB diversity (richness and community structure). Within sites, tree species influence AOB diversity. The absence of a relation between AOB richness and nitrification points to a possibly role of AOB abundance, phenotypic plasticity or the implication of ammonia oxidizing archaea.