IntroductionPesticides are widely used in agriculture, yet their non-target effects on soil microbial communities remain poorly understood. This study investigates the short-term impact of five herbicides and three fungicides used for the protection of legumes on the composition and diversity of soil bacteria, with special focus on rhizobia.MethodsUsing three distinct soils from ecologically maintained fields, we assessed changes in bacterial communities and total bacterial abundance in response to different active substances under controlled conditions, 2 weeks after pesticide application. Bacterial diversity was analyzed by amplifying and sequencing the V4 region of the 16S rRNA gene via Illumina paired-end amplicon sequencing. Real-time PCR was used to assess total abundance of bacteria.ResultsOur results show that pesticide effects on bacteria are highly context-dependent, influenced significantly by soil and pH. Significant changes in bacterial diversity were detected only in one soil, whereas in another soil no significant differences among individual pesticides and the untreated control were found. In this soil, particularly the CORUM and pendimethalin-based products, Stomp 400 SC and Sharpen 40 SC, consistently reduced bacterial diversity, while some pesticides had a neutral effect. Rhizobial diversity remained largely unaffected, suggesting greater resilience compared to general bacterial communities. Regarding bacterial community composition, only some pesticides significantly affected bacterial community structure in each soil, and the pesticides showing this effect differed among soils. Redundancy analysis revealed that pH was a stronger driver of bacterial community structure than soil type or pesticide identity, explaining over 60% of community variability.ConclusionThese findings highlight the complex interactions between pesticides, soil characteristics, and microbial communities. Our results support considering soil pH when selecting pesticides to support sustainable soil management and minimize ecological disruption.

Biofilm and planktonic prokaryotic communities were studied using a glass fibre filter as trapping material immersed in field soil at different times of the year (January, April, July, September) and incubated there for different periods (3, 6, 9, 12 months). The composition of biofilm and planktonic communities fluctuated over time, likely shaped by succession processes and varying environmental factors. This highlights soil biofilms as dynamic structures whose microbial community differs from that of soil plankton. Additionally, quantification of the biofilm-to-plankton 16S rRNA gene copy number ratio indicated that soil prokaryotes occur mainly as biofilm components.

Sarcosphaera (Ascomycota: Pezizales) is a genus of rare spring fungi, currently encompassing a single species, S. coronaria, which is red-listed in many European countries. The genus is characterized by large bright-colored fruit-bodies and high arsenic content. This study investigates the genetic diversity across Europe and North America. Using integrated molecular techniques, including ITS and 28S rDNA sequencing, we aimed to resolve the taxonomic ambiguities and assess their arsenic hyperaccumulation capabilities, a rare trait among fungi. We identified the presence of two species in the Western Palearctic (S. coronaria, S. crassa) and at least 12 additional lineages in North America, indicating a more complex taxonomic structure than previously recognized. To consolidate the taxonomy and nomenclature of these fungi, we selected reference specimens for S. coronaria and S. crassa and discussed their morphological and ecological differences. Two genera, Caulocarpa (C. montana) and Hydnotryopsis (H. setchellii), a genus comprising hypogeous fungi, are newly placed in the genus Sarcosphaera. Sarcosphaera is thus another genus where the rapid evolution of hypogeous forms from their above-ground ancestors took place. Furthermore, two new Sarcosphaera species, S. columbiana and S. pacifica, are described from the Pacific Northwest. Investigation of arsenic content of Sarcosphaera species revealed that arsenic mass fractions in ascocarps vary significantly between species and even within species across different geographical locations. Our findings have important implications for the taxonomy and conservation of the S. coronaria species complex and also point to future directions for further research in the taxonomy, geomycology, and toxicology of this group of fungi.

Arbuscular mycorrhizal fungi (AMF) improve plant growth and may be useful in maintaining and even restoring soil. However, data on the latter function are sparse and only indirect, which is especially true for conventional management conditions with adequate nutrient availability. Our study focused on utilizing the prevalent AMF species, Funneliformis mosseae, to enhance Glycine max production, while also exploring its partly explored impact on soil aggregation. Working in greenhouse conditions, we examined whether, in a nutrient-sufficient environment, AMF would improve crop biomass accumulation and nutrition, as well as the stability of soil aggregates (SAS). We also looked for a synergistic effect of dual inoculation using AMF and symbiotic rhizobium. Plants were or were not inoculated with AMF or Bradyrhizobium japonicum in a two-factorial design. AMF inoculation increased soybean biomass, but AMF inoculation had no impact on P and N input to the shoots. Mycorrhiza did not affect either glomalin abundance or SAS. All the impacts were, however, independent from rhizobial inoculation, which was ineffective in this nutrient-available environment. Our assay suggests that arbuscular mycorrhiza may have a positive effect on soybean growth even under conventional management with adequate nutrition. The positive effects of AMF on soybean growth, together with the fact that AMF generally do not thrive in good nutrient availability, should be taken into account when planning mineral fertilization levels.

Arbuscular mycorrhizal fungi (AMF) provide crucial support for the establishment of plants in novel environments. We hypothesized that the OTU/genus richness and diversity of soil- and root-associated AMF associated with alien plant species in their exotic ranges are lower than those in their native ranges. We examined the root-associated and soil-dwelling AMF of 11 invasive plant species in their native and exotic ranges in the United States and Europe by DNA sequencing of the ITS2 locus. Examined root-associated AMF assemblages were simplified, which manifested as the loss of several AMF genera in the exotic ranges of the plants. These fungal assemblages were also characterized by greater dominance and simplification of the fungal assemblages. The dominant fungal genera were present regardless of whether their host plants were in their native or exotic ranges. Interestingly, both the native and invaded soils hosted diverse local AMF assemblages. Therefore, alien plant invasions were not limited to soils with low AMF diversity. Some AMF taxa could be context-dependent passengers rather than drivers of alien plant invasions. Further studies should identify functions of AMF missing or less abundant in roots of plants growing in exotic ranges.

Microbial community of the soil can be subdivided into a planktonic community of motile organisms and a community of sessile organisms, including those constituting biofilms on physical surfaces of different texture. Biofilm communities on surfaces of organic particles represent a crucial factor involved in decomposition of plant litter and soil particulate organic matter in general. In spite of their great ecological importance, the composition of soil biofilm communities is only scarcely studied due to methodological problems connected with distinguishing biofilm inhabitants from other soil organisms.

Biofilms are dynamic structures constituted by microorganisms that grow and die, and understanding these processes may be crucial to control biofilm development in various environments. Assuming a generally accepted first order decay kinetics of biofilm mass in time, mean residence time can be calculated. Using the initial labeling of the biofilm by 13C stable isotope, we were able to determine the residence time of the carbon in physiologically active biofilms. Our data indicate that the residence time is strongly affected by nutrition and differs substantially between biofilms formed by different bacterial isolates. Moreover, the biofilm formed from mixed soil inocula showed almost the same carbon residence time as the biofilms formed from both soil inocula applied separately. This does not indicate the existence of dramatic incompatibility between members of two interacting microbial communities. In the situation when the established biofilm biomass undergoes continuous replacement by newly appearing cells, the complex biofilm admit reluctantly the newly arriving microorganisms as components of the existing community. Our study represents a new insight into the biofilm dynamics in vitro.

Invasive plants often pose serious threats to the natural biodiversity of invaded ecosystems and in this way are likely to alter ecosystem services. This applies to arbuscular mycorrhizal (AM) fungi, in which the invaders have been reported to modify community structure, which facilitates their further intrusion. Information as to the impact of such invasion on AM fungal communities is insufficient and therefore needed. In particular, little is known about how AM fungal communities shift in response to individual invasive species. To ascertain whether invasion changes the structure of indigenous AM fungal communities, we examined changes in AM fungal community composition and diversity in soil and in roots of native neighboring plants in response to incursion of five invasive plant species from the family Asteraceae: Conyza canadensis, Erigeron annuus, Echinops sphaerocephalus, Solidago canadensis, and Symphyotrichum novi-belgii. We found that invasions of tested invasive plant species altered composition of the AM fungal community and reduced the diversity of AM fungi in soil and in the roots of some native plants. Statistical significance of the invasions' effects depended on composition of AM fungal communities in roots of the native plant species and/or site and was not connected with changes in soil parameters. Our results confirm the notable influence of plant invasion on indigenous AM fungal biodiversity and the need for further study in various environmental conditions.

Arbuscular mycorrhizal (AM) fungi establish symbiotic associations with many plant species, transferring significant amounts of soil nutrients such as phosphorus to plants and receiving photosynthetically fixed carbon in return. Functioning of AM symbiosis is thus based on interaction between two living partners. The importance of dead AM fungal biomass (necromass) in ecosystem processes remains unclear. Here, we applied either living biomass or necromass (0.0004 potting substrate weight percent) of monoxenically produced AM fungus (Rhizophagus irregularis) into previously sterilized potting substrate planted with Andropogon gerardii. Plant biomass production significantly improved in both treatments as compared to non-amended controls. Living AM fungus, in contrast to the necromass, specifically improved plant acquisition of nutrients normally supplied to the plants by AM fungal networks, such as phosphorus and zinc. There was, however, no difference between the two amendment treatments with respect to plant uptake of other nutrients such as nitrogen and/or magnesium, indicating that the effect on plants of the AM fungal necromass was not primarily nutritional. Plant growth stimulation by the necromass could thus be either due to AM fungal metabolites directly affecting the plants, indirectly due to changes in soil/root microbiomes or due to physicochemical modifications of the potting substrate. In the necromass, we identified several potentially bioactive molecules. We also provide experimental evidence for significant differences in underground microbiomes depending on the amendment with living or dead AM fungal biomass. This research thus provides the first glimpse into possible mechanisms responsible for observed plant growth stimulation by the AM fungal necromass.

Biofilm formation is a typical life strategy used by microorganisms populating acidic water systems. The same strategy might be used by microbes in highly acidic soils that are, however, neglected in this regard. In the present study, the microbial community in such highly acidic soil in the Soos National Nature Reserve (Czech Republic) has been investigated using high-throughput DNA sequencing and the organisms associated with biofilm life mode and those preferring planktonic life were distinguished using the biofilm trap technique. Our data show the differences between biofilm and planktonic microbiota fraction, although the majority of the organisms were capable of using both life modes. The by far most abundant prokaryotic genus was Acidiphilium and fungi were identified among the most abundant eukaryotic elements in biofilm formations. On the other hand, small flagellates from diverse taxonomical groups predominated in plankton. The application of cellulose amendment as well as the depth of sampling significantly influenced the composition of the detected microbial community.

The microbial biofilms are ubiquitous in nature and represent important biological entities that affect various aspects of human life. As such, they attracted considerable attention during last decades, with the factors affecting the biofilm development being among the frequently studied topics. In our work, the biofilm was cultivated on the surface of polypropylene fibers in a nutrient medium inoculated by the suspension of two unsterile soils. The effects of ionic strength and valence of salt on the amount of the produced biofilm and on composition of biofilm microbial communities were investigated. The effect of valence was significant in some OTUs: Arthrobacter/Pseudarthrobacter/Paenarthrobacter and Bacillus with positive response to monovalent salt (KCl) and Streptomyces, Lysinibacillus, Pseudomonas, and Ensifer with positive response to divalent salt (MgSO4). The significant preference for a certain concentration of salts was observed in the case of OTUs Agrobacterium, Bacillus (both 100 mM), and Brevundimonas (30 mM). A new quantification method based on measuring of oxidizable organic carbon in biofilm biomass, based on dichromate oxidation, was used. We compared the results obtained using this method with results of crystal violet destaining and measuring of extracted DNA concentration as proxies of the biofilm biomass. The dichromate oxidation is simple, inexpensive, and fast, and our results show that it may be more sensitive than crystal violet destaining. The highest biomass values tended to associate with high concentrations of the divalent salt. This trend was not observed in treatments where the monovalent salt was added. Our data confirm the importance of inorganic ions for biofilm composition and biomass accumulation.

Fungi from extreme environments, including acidophilic ones, belong to biotechnologically most attractive organisms. They can serve as a source of enzymes and metabolites with potentially uncommon properties and may actively participate within bioremediation processes. In respect of their biotechnological potential, extremophilic fungi are mostly studied as individual species. Nevertheless, microorganisms rarely live separately and they form biofilms instead. Living in biofilms is the most successful life strategy on the Earth and the biofilm is the most abundant form of life in extreme environments including highly acidic ones. Compared to bacterial fraction, fungal part of acidophilic biofilms represents a largely unexplored source of organisms with possible use in biotechnology and especially data on biofilms of highly acidic soils are missing. The functioning of the biofilm results from interactions between organisms whose metabolic capabilities are efficiently combined. When we look on acidophilic fungi and their biotechnological potential we should take this fact into account as well. The practical problem to be resolved in connection with extensive studies of exploitable properties and abilities of acidophilic fungi is the methodology of isolation of strains from the nature. In this respect, novel isolation techniques should be developed.

Despite the crucial importance of arbuscular mycorrhizal fungi (AMF) for numerous processes within terrestrial ecosystems, knowledge of the determinants of AMF community structure still is limited, mainly because of the limited scope of the available individual case studies which often only include a few environmental variables. Here, we describe the AMF diversity of mid-European meadows (mown or regularly cut grasslands, or recently abandoned lands where grasslands established spontaneously) within a considerably heterogeneous landscape over a scale of several hundred kilometers with regard to macroclimatic, microclimatic, and soil parameters. We include data describing the habitat (including vegetation type), geography, and climate, and test their contribution to the structure of the AMF communities at a regional scale. We amplified and sequenced the ITS 2 region of the ribosomal DNA operon of the AMF from soil samples using nested PCR and Illumina pair-end amplicon sequencing. Habitat (especially soil pH) and geographical parameters (spatial distance, altitude, and longitude) were the main determinants of the structure of the AMF communities in the meadows at a regional scale, with the abundance of genera Septoglomus , Paraglomus , Archaeospora , Funneliformis , and Dominikia driving the main response. The effects of climate and vegetation type were not significant and were mainly encompassed within the geography and/or soil pH effects. This study illustrates how important it is to have a large set of environmental metadata to compare the importance of different factors influencing the AMF community structure at large spatial scales.

Amanita strobiliformis (European Pine Cone Lepidella) is an ectomycorrhizal fungus of the Amanitaceae family known to hyperaccumulate Ag in the sporocarps. Two populations (ecotypes) of A. strobiliformis collected from two urban forest plantations in Prague, Czech Republic, were investigated. The concentrations of Ag, Cu, Cd, and Zn were determined in the mushrooms. The metal mobility and fractionation in the soils was investigated by single extractions and sequential extraction. The soil distribution of A. strobiliformis mycelium was assessed by quantitative polymerase chain reaction (qPCR). The metal uptake from the soil into the mushroom sporocarps was traced by Pb isotopic fingerprinting. The findings suggested that A. strobiliformis (i) accumulates primarily Ag from the topsoil layer (circa 12cm deep) and (ii) accumulates Ag associated with the “reducible soil fraction”. The concentrations of all metals, particularly Ag and Cu, were significantly higher in the A. strobiliformis sporocarps from one of the investigated sites (Klíčov). The elevated concentrations of Ag in the sporocarps from Klíčov can possibly be attributed to the higher Ag content in the topsoil layer found at this site. However, the simultaneously elevated concentrations of Cu in A. strobiliformis from Klíčov cannot be explained by the differences in the geochemical background and should be attributed to biological factors.

Biofilms, the communities of sessile microbial forms, are hotspots of biological activity that coexist in soil together with free-living (planktonic) biota. Sessile and planktonic microbial communities may constitute functionally different groups of organisms with different roles in interactions with organic matter and plants. Nevertheless, soil microbiomes are regularly analyzed without distinguishing biofilm inhabitants and planktonic organisms. Consequently, basic information regarding taxa constituting both communities is severely lacking, which limits the understanding of the basic structure of soil microbiome and consequently also of the microbiome functioning.

Establishment of nonmycorrhizal controls is a "classic and recurrent theme" in mycorrhizal research. For decades, authors reported mycorrhizal plant growth/nutrition as compared to various nonmycorrhizal controls. In such studies, uncertainties remain about which nonmycorrhizal controls are most appropriate and, in particular, what effects the control inoculations have on substrate and root microbiomes. Here, different types of control and mycorrhizal inoculations were compared with respect to plant growth and nutrition, as well as the structure of root and substrate microbiomes, assessed by next-generation sequencing. We compared uninoculated ("absolute") control to inoculation with blank pot culture lacking arbuscular mycorrhizal fungi, filtrate of that blank inoculum, and filtrate of complex pot-produced mycorrhizal inoculum. Those treatments were compared to a standard mycorrhizal treatment, where the previously sterilized substrate was inoculated with complex pot-produced inoculum containing Rhizophagus irregularis SYM5. Besides this, monoxenically produced inoculum of the same fungus was applied either alone or in combination with blank inoculum. The results indicate that the presence of mycorrhizal fungus always resulted in stimulation of Andropogon gerardii plant biomass as well as in elevated phosphorus content of the plants. The microbial (bacterial and fungal) communities developing in the differently inoculated treatments, however, differed substantially from each other and no control could be obtained comparable with the treatment inoculated with complex mycorrhizal inoculum. Soil microorganisms with significant biological competences that could potentially contribute to the effects of the various inoculants on the plants were detected in roots and in plant cultivation substrate in some of the treatments.

Biochar has been heralded as a multipurpose soil amendment to sustainably increase soil fertility and crop yields, affect soil hydraulic properties, reduce nutrient losses, and sequester carbon. Some of the most spectacular results of biochar (and organic nutrient) inputs are the terra preta soils in the Amazon, dark anthropogenic soils with extremely high fertility sustained over centuries. Such soil improvements have been particularly difficult to achieve on a short run, leading to speculations that biochar may need to age (weather) in soil to show its best. Further, interaction of biochar with arbuscular mycorrhizal fungi (AMF), important root symbionts of a great majority of terrestrial plants including most agricultural crops, remains little explored. To study the effect of aged biochar on highly mycotrophic Andropogon gerardii plants and their associated AMF, we made use of softwood biochar, collected from a historic charcoal burning site. This biochar (either untreated or chemically activated, the latter serving as a proxy for freshly prepared biochar) was added into two agricultural soils (acid or alkaline), and compared to soils without biochar. These treatments were further crossed with inoculation with a synthetic AMF community to address possible interactions between biochar and the AMF. Biochar application was generally detrimental for growth and mineral nutrition of our experimental plants, but had no effect on the extent of their root colonized by the AMF, nor did it affect composition of their root-borne AMF communities. In contrast, biochar affected development of two out of five AMF (Claroideoglomus and Funneliformis) in the soil. Establishment of symbiosis with AMF largely mitigated biochar-induced suppression of plant growth and mineral nutrition, mainly by improving plant acquisition of phosphorus. Both mycorrhizal and non-mycorrhizal plants grew well in the acid soil without biochar application, whereas non-mycorrhizal plants remained stunted in the alkaline soils under all situations (with or without biochar). These different and strong effects indicate that response of plants to biochar application are largely dependent on soil matrix and also on microbes such as AMF, and call for further research to enable qualified predictions of the effects of different biochar applications on field-grown crops and soil processes.

Arbuscular mycorrhizal (AM) fungi can significantly contribute to plant nitrogen (N) uptake from complex organic sources, most likely in concert with activity of soil saprotrophs and other microbes releasing and transforming the N bound in organic forms. Here, we tested whether AM fungus (Rhizophagus irregularis) extraradical hyphal networks showed any preferences towards certain forms of organic N (chitin of fungal or crustacean origin, DNA, clover biomass, or albumin) administered in spatially discrete patches, and how the presence of AM fungal hyphae affected other microbes. By direct 15N labeling, we also quantified the flux of N to the plants (Andropogon gerardii) through the AM fungal hyphae from fungal chitin and from clover biomass. The AM fungal hyphae colonized patches supplemented with organic N sources significantly more than those receiving only mineral nutrients, organic carbon in form of cellulose, or nothing. Mycorrhizal plants grew 6.4-fold larger and accumulated, on average, 20.3-fold more 15N originating from the labeled organic sources than their nonmycorrhizal counterparts. Whereas the abundance of microbes (bacteria, fungi, or Acanthamoeba sp.) in the different patches was primarily driven by patch quality, we noted a consistent suppression of the microbial abundances by the presence of AM fungal hyphae. This suppression was particularly strong for ammonia oxidizing bacteria. Our results indicate that AM fungi successfully competed with the other microbes for free ammonium ions and suggest an important role for the notoriously understudied soil protists to play in recycling organic N from soil to plants via AM fungal hyphae.

Although arbuscular mycorrhizal symbiosis is common in many plants with either C3 or C4 photosynthesis, it remains poorly understood whether photosynthesis type has any significant impact on carbon (C) fluxes in mycorrhizal plants. Thus, we compared mycorrhizal and non-mycorrhizal (NM) plants belonging to Panicum bisulcatum (C3) to its congeneric P. maximum (C4).