Multiple anthropogenic pressures eliminate the effects of soil microbial diversity on ecosystem functions in experimental microcosms.

Biodiversity is crucial for the provision of ecosystem functions. However, ecosystems are now exposed to a rapidly growing number of anthropogenic pressures, and it remains unknown whether biodiversity can still promote ecosystem functions under multifaceted pressures. Here we investigated the effects of soil microbial diversity on soil functions and properties when faced with an increasing number of simultaneous global change factors in experimental microcosms. Higher soil microbial diversity had a positive effect on soil functions and properties when no or few (i.e., 1-4) global change factors were applied, but this positive effect was eliminated by the co-occurrence of numerous global change factors. This was attributable to the reduction of soil fungal abundance and the relative abundance of an ecological cluster of coexisting soil bacterial and fungal taxa. Our study indicates that reducing the number of anthropogenic pressures should be a goal in ecosystem management, in addition to biodiversity conservation.

Fungal response to abruptly or gradually delivered antifungal agent amphotericin B is growth stage dependent.

Anthropogenic disturbances pose a multitude of novel challenges to ecosystems. While many experiments have tested effects using abrupt treatment applications, most environmental changes in fact are gradual. Since ecosystem responses might be highly dependent on the temporal nature of stressors, it is crucial to differentiate the effects of abrupt vs gradual treatment application. Antifungal agents, which are widely used in disease control both for humans and in agriculture, are becoming a new class of environmental contaminants. In this study, we examined the effect of a sub-lethal application of one antifungal agent, amphotericin B. We applied different rates of delivery, e.g. gradual and abrupt, and monitored biomass and sporulation of the model fungus Neurospora crassa in a batch culture. Our results demonstrate that: (i) the effect size difference between abrupt and gradual treatments is fungal growth stage dependent and (ii) the gradual treatment clearly had a higher sporulation level compared with all types of abrupt treatments. Our findings highlight the importance of considering the rate of change in environmental change research and point to a new research direction for future global change studies. Furthermore, our results also have important implications for avoiding treatment-induced spore production in agriculture and medical practise.

It Takes Two to Tango: A Bacterial Biofilm Provides Protection against a Fungus-Feeding Bacterial Predator.

Fungus-bacterium interactions are widespread, encompass multiple interaction types from mutualism to parasitism, and have been frequent targets for microbial inoculant development. In this study, using in vitro systems combined with confocal laser scanning microscopy and real-time quantitative PCR, we test whether the nitrogen-fixing bacterium Kosakonia radicincitans can provide protection to the plant-beneficial fungus Serendipita indica, which inhabits the rhizosphere and colonizes plants as an endophyte, from the fungus-feeding bacterium Collimonas fungivorans. We show that K. radicincitans can protect fungal hyphae from bacterial feeding on solid agar medium, with probable mechanisms being quick hyphal colonization and biofilm formation. We furthermore find evidence for different feeding modes of K. radicincitans and C. fungivorans, namely "metabolite" and "hyphal feeding", respectively. Overall, we demonstrate, to our knowledge, the first evidence for a bacterial, biofilm-based protection of fungal hyphae against attack by a fungus-feeding, bacterial predator on solid agar medium. Besides highlighting the importance of tripartite microbial interactions, we discuss implications of our results for the development and application of microbial consortium-based bioprotectants and biostimulants.

A non-invasive soil-based setup to study tomato root volatiles released by healthy and infected roots.

The role of root exudates in mediating plant-microbe interactions has been well documented. However, the function of volatile organic compounds (VOCs) emitted by plant roots has only recently begun to attract attention. This newly recognized relevance of belowground VOCs has so far mostly been tested using systems limited to a two-compartment Petri-dish design. Furthermore, many of the plant-microbe interaction studies have only investigated the effects of microbial VOCs on plant growth. Here, we go two steps further. First we investigated the volatile profile of healthy and pathogen (Fusarium oxysporum) infected tomato roots grown in soil. We then used a unique soil-based olfactometer-choice assay to compare the migration pattern of four beneficial bacteria (Bacillus spp.) towards the roots of the tomato plants. We demonstrate that the blend of root-emitted VOCs differs between healthy and diseased plants. Our results show that VOCs are involved in attracting bacteria to plant roots.

Soil Saprobic Fungi Differ in Their Response to Gradually and Abruptly Delivered Copper.

The overwhelming majority of studies examining environmental change deliver treatments abruptly, although, in fact, many important changes are gradual. One example of a gradually increasing environmental stressor is heavy metal contamination. Essential heavy metals, such as copper, play an important role within cells of living organisms but are toxic at higher concentrations. In our study, we focus on the effects of copper pollution on filamentous soil fungi, key players in terrestrial ecosystem functioning. We hypothesize that fungi exposed to gradually increasing copper concentrations have higher chances for physiological acclimation and will maintain biomass production and accumulate less copper, compared to fungi abruptly exposed to the highest copper concentration. To test this hypothesis, we conducted an experiment with 17 fungal isolates exposed to gradual and abrupt copper addition. Contrary to our hypothesis, we find diverse idiosyncratic responses, such that for many fungi gradually increasing copper concentrations have more severe effects (stronger growth inhibition and higher copper accumulation) than an abrupt increase. While a number of environmental change studies have accumulated evidence based on the magnitude of changes, the results of our study imply that the rate of change can be an important factor to consider in future studies in ecology, environmental science, and environmental management.

The interaction between Rhizoglomus irregulare and hyphae attached phosphate solubilizing bacteria increases plant biomass of Solanum lycopersicum.

The synergistic interaction between arbuscular mycorrhizal fungi (AMF) and phosphate solubilizing bacteria (PSB) can enhance growth and phosphorous uptake in plants. Since PSBs are well known hyphal colonizers we sought to understand this physical interaction and exploit it in order to design strategies for the application of a combined microbial inoculum. Phosphate-solubilizing bacteria strongly attached to the hyphae of Rhizoglomus irregulare were isolated using a two compartment system (root and hyphal compartments), which were separated by a nylon mesh through which AMF hyphae could pass but not plant roots. Allium ampeloprasum (Leek) was used as the host plant inoculated with R. irregulare. A total of 128 bacteria were isolated, of which 12 showed stable phosphate solubilizing activity. Finally, three bacteria belonging to the genus Pseudomonas showed the potential for inorganic and organic phosphate mobilization along with other plant growth promoting traits. These PSBs were further evaluated for their functional characteristics and their interaction with AMF. The impact of single or co-inoculations of the selected bacteria and AMF on Solanum lycopersicum was tested and we found that plants inoculated with the combination of fungus and bacteria had significantly higher plant biomass compared to single inoculations, indicating synergistic activities of the bacterial-fungal consortium.

Mimicking climate warming effects on Alaskan soil microbial communities via gradual temperature increase.

Climate change can trigger shifts in community structure and may therefore pose a severe threat to soil microbial communities, especially in high northern latitudes such as the Arctic. Arctic soils are covered by snow and ice throughout most of the year. This insulation shields them from high temperature variability and low surface temperatures. If this protective layer thaws, these soils are predicted to warm up at 1.5x to 4x the rate of other terrestrial biomes. In this study, we sampled arctic soils from sites with different elevations in Alaska, incubated them for 5 months with a simulated, gradual or abrupt temperature increase of +5 °C, and compared bacterial and fungal community compositions after the incubation. We hypothesized that the microbial communities would not significantly change with a gradual temperature treatment, whereas an abrupt temperature increase would decrease microbial diversity and shift community composition. The only differences in community composition that we observed were, however, related to the two elevations. The abrupt and gradual temperature increase treatments did not change the microbial community composition as compared to the control indicating resistance of the microbial community to changes in temperature. This points to the potential importance of microbial dormancy and resting stages in the formation of a "buffer" against elevated temperatures. Microbial resting stages might heavily contribute to microbial biomass and thus drive the responsiveness of arctic ecosystems to climate change.

Myristate and the ecology of AM fungi: significance, opportunities, applications and challenges.

A recent study by Sugiura and coworkers reported the non-symbiotic growth and spore production of an arbuscular mycorrhizal (AM) fungus, Rhizophagus irregularis, when the fungus received an external supply of certain fatty acids, myristates (C:14). This discovery follows the insight that AM fungi receive fatty acids from their hosts when in symbiosis. If this result holds up and can be repeated under nonsterile conditions and with a broader range of fungi, it has numerous consequences for our understanding of AM fungal ecology, from the level of the fungus, at the plant community level, and to functional consequences in ecosystems. In addition, myristate may open up several avenues from a more applied perspective, including improved fungal culture and supplementation of AM fungi or inoculum in the field. We here map these potential opportunities, and additionally offer thoughts on potential risks of this potentially new technology. Lastly, we discuss the specific research challenges that need to be overcome to come to an understanding of the potential role of myristate in AM ecology.

Increasing Temperature and Microplastic Fibers Jointly Influence Soil Aggregation by Saprobic Fungi.

Microplastic pollution and increasing temperature have potential to influence soil quality; yet little is known about their effects on soil aggregation, a key determinant of soil quality. Given the importance of fungi for soil aggregation, we investigated the impacts of increasing temperature and microplastic fibers on aggregation by carrying out a soil incubation experiment in which we inoculated soil individually with 5 specific strains of soil saprobic fungi. Our treatments were temperature (ambient temperature of 25°C or temperature increased by 3°C, abruptly versus gradually) and microplastic fibers (control and 0.4% w/w). We evaluated the percentage of water stable aggregates (WSA) and hydrolysis of fluorescein diacetate (FDA) as an indicator of fungal biomass. Microplastic fiber addition was the main factor influencing the WSA, decreasing the percentage of WSA except in soil incubated with strain RLCS 01, and mitigated the effects of temperature or even caused more pronounced decrease in WSA under increasing temperature. We also observed clear differences between temperature change patterns. Our study shows that the interactive effects of warming and microplastic fibers are important to consider when evaluating effects of global change on soil aggregation and potentially other soil processes.

Trait Differentiation within the Fungus-Feeding (Mycophagous) Bacterial Genus Collimonas.

The genus Collimonas consists of facultative, fungus-feeding (mycophagous) bacteria. To date, 3 species (C. fungivorans, C. pratensis and C. arenae) have been described and over 100 strains have been isolated from different habitats. Functional traits of Collimonas bacteria that are potentially involved in interactions with soil fungi mostly negatively (fungal inhibition e.g.), but also positively (mineral weathering e.g.), affect fungal fitness. We hypothesized that variation in such traits between Collimonas strains leads to different mycophagous bacterial feeding patterns. We investigated a) whether phylogenetically closely related Collimonas strains possess similar traits, b) how far phylogenetic resolution influences the detection of phylogenetic signal (possession of similar traits by related strains) and c) if there is a pattern of co-occurrence among the studied traits. We measured genetically encoded (nifH genes, antifungal collimomycin gene cluster e.g.) as well as phenotypically expressed traits (chitinase- and siderophore production, fungal inhibition and others) and related those to a high-resolution phylogeny (MLSA), constructed by sequencing the housekeeping genes gyrB and rpoB and concatenating those with partial 16S rDNA sequences. Additionally, high-resolution and 16S rDNA derived phylogenies were compared. We show that MLSA is superior to 16SrDNA phylogeny when analyzing trait distribution and relating it to phylogeny at fine taxonomic resolution (a single bacterial genus). We observe that several traits involved in the interaction of collimonads and their host fungus (fungal inhibition e.g.) carry phylogenetic signal. Furthermore, we compare Collimonas trait possession with sister genera like Herbaspirillum and Janthinobacterium.

Methods for Baiting and Enriching Fungus-Feeding (Mycophagous) Rhizosphere Bacteria.

Mycophagous soil bacteria are able to obtain nutrients from living fungal hyphae. However, with exception of the soil bacterial genus Collimonas, occurrence of this feeding strategy has not been well examined. Evaluation of the importance of mycophagy in soil bacterial communities requires targeted isolation methods. In this study, we compared two different approaches to obtain mycophagous bacteria from rhizospheric soil. A short-term method based on baiting for bacteria that can rapidly adhere to fungal hyphae and a long-term method based on the enrichment of bacteria on fungal hyphae via repeated transfer. Hyphae-adhering bacteria were isolated, identified by 16S rDNA sequencing and tested for antifungal activity and the ability to feed on fungi as the sole source of carbon. Both methods yielded a range of potentially mycophagous bacterial isolates with little phylogenetic overlap. We also found indications for feeding preferences among the potentially mycophagous bacteria. Our results indicate that mycophagy could be an important growth strategy for rhizosphere bacteria. To our surprise, we found several potential plant pathogenic bacteria among the mycophagous isolates. We discuss the possible benefits that these bacteria might gain from colonizing fungal hyphae.