Subarctic winter warming promotes soil microbial resilience to freeze-thaw cycles and enhances the microbial carbon use efficiency.

Climate change is predicted to cause milder winters and thus exacerbate soil freeze-thaw perturbations in the subarctic, recasting the environmental challenges that soil microorganisms need to endure. Historical exposure to environmental stressors can facilitate the microbial resilience to new cycles of that same stress. However, whether and how such microbial memory or stress legacy can modulate microbial responses to cycles of frost remains untested. Here, we conducted an in situ field experiment in a subarctic birch forest, where winter warming resulted in a substantial increase in the number and intensity of freeze-thaw events. After one season of winter warming, which raised mean surface and soil (-8 cm) temperatures by 2.9 and 1.4°C, respectively, we investigated whether the in situ warming-induced increase in frost cycles improved soil microbial resilience to an experimental freeze-thaw perturbation. We found that the resilience of microbial growth was enhanced in the winter warmed soil, which was associated with community differences across treatments. We also found that winter warming enhanced the resilience of bacteria more than fungi. In contrast, the respiration response to freeze-thaw was not affected by a legacy of winter warming. This translated into an enhanced microbial carbon-use efficiency in the winter warming treatments, which could promote the stabilization of soil carbon during such perturbations. Together, these findings highlight the importance of climate history in shaping current and future dynamics of soil microbial functioning to perturbations associated with climate change, with important implications for understanding the potential consequences on microbial-mediated biogeochemical cycles.

Heat wave-induced microbial thermal trait adaptation and its reversal in the Subarctic.

Climate change predictions suggest that arctic and subarctic ecosystems will be particularly affected by rising temperatures and extreme weather events, including severe heat waves. Temperature is one of the most important environmental factors controlling and regulating microbial decomposition in soils; therefore, it is critical to understand its impact on soil microorganisms and their feedback to climate warming. We conducted a warming experiment in a subarctic birch forest in North Sweden to test the effects of summer heat waves on the thermal trait distributions that define the temperature dependences for microbial growth and respiration. We also determined the microbial temperature dependences 10 and 12 months after the heat wave simulation had ended to investigate the persistence of the thermal trait shifts. As a result of warming, the bacterial growth temperature dependence shifted to become warm-adapted, with a similar trend for fungal growth. For respiration, there was no shift in the temperature dependence. The shifts in thermal traits were not accompanied by changes in α- or ß-diversity of the microbial community. Warming increased the fungal-to-bacterial growth ratio by 33% and decreased the microbial carbon use efficiency by 35%, and both these effects were caused by the reduction in moisture the warming treatments caused, while there was no evidence that substrate depletion had altered microbial processes. The warm-shifted bacterial thermal traits were partially restored within one winter but only fully recovered to match ambient conditions after 1 year. To conclude, a summer heat wave in the Subarctic resulted in (i) shifts in microbial thermal trait distributions; (ii) lower microbial process rates caused by decreased moisture, not substrate depletion; and (iii) no detectable link between the microbial thermal trait shifts and community composition changes.

Variation in Temperature Dependences across Europe Reveals the Climate Sensitivity of Soil Microbial Decomposers.

Temperature is a major determinant of biological process rates, and microorganisms are key regulators of ecosystem carbon (C) dynamics. Temperature controls microbial rates of decomposition, and thus warming can stimulate C loss, creating positive feedback to climate change. If trait distributions that define temperature relationships of microbial communities can adapt to altered temperatures, they could modulate the strength of this feedback, but if this occurs remains unclear. In this study, we sampled soils from a latitudinal climate gradient across Europe. We established the temperature relationships of microbial growth and respiration rates and used these to investigate if and with what strength the community trait distributions for temperature were adapted to their local environment. Additionally, we sequenced bacterial and fungal amplicons to link the variance in community composition to changes in temperature traits. We found that microbial temperature trait distributions varied systematically with climate, suggesting that an increase in mean annual temperature (MAT) of 1°C will result in warm-shifted microbial temperature trait distributions equivalent to an increase in temperature minimum (Tmin) of 0.20°C for bacterial growth, 0.07°C for fungal growth, and 0.10°C for respiration. The temperature traits for bacterial growth were thus more responsive to warming than those for respiration and fungal growth. The microbial community composition also varied with temperature, enabling the interlinkage of taxonomic information with microbial temperature traits. Our work shows that the adaptation of microbial temperature trait distributions to a warming climate will affect the C-climate feedback, emphasizing the need to represent this to capture the microbial feedback to climate change. IMPORTANCE One of the largest uncertainties of global warming is if the microbial decomposer feedback will strengthen or weaken soil C-climate feedback. Despite decades of research effort, the strength of this feedback to warming remains unknown. We here present evidence that microbial temperature relationships vary systematically with environmental temperatures along a climate gradient and use this information to forecast how microbial temperature traits will create feedback between the soil C cycle and climate warming. We show that the current use of a universal temperature sensitivity is insufficient to represent the microbial feedback to climate change and provide new estimates to replace this flawed assumption in Earth system models. We also demonstrate that temperature relationships for rates of microbial growth and respiration are differentially affected by warming, with stronger responses to warming for microbial growth (soil C formation) than for respiration (C loss from soil to atmosphere), which will affect the atmosphere-land C balance.

Can heavy metal pollution induce bacterial resistance to heavy metals and antibiotics in soils from an ancient land-mine?

Microbial resistance to antibiotics is a growing challenge to human health. Recent evidence has indicated that antibiotic resistance can be co-selected for by exposure to heavy metals in agricultural soils. It remains unknown if this is a concern in other environments contaminated by metals. We here investigated soil microbial activities, composition and tolerance to heavy metals and antibiotics in a mining soil survey. We found that microbial respiration, growth, and biomass were affected by available metal concentrations. Most of the variation in microbial PLFA composition was explained by differences in heavy metal and pH. Additionally, pollution-induced bacterial community tolerance to toxicants including Cu, Pb, Zn, tetracycline and vancomycin was determined. Although only bacterial tolerance to Pb increased with higher levels of metals, the links between bacterial metal tolerance and soil metal concentrations were clear when considered together with previously published reports, suggesting that bacterial metal tolerance were universally elevated in the surveyed soils. The induced levels of heavy metal tolerance coincided with elevated levels of tolerance to vancomycin, but not to tetracycline. Our study showed that heavy metals can co-select for resistance to clinically important antibiotics also in ecosystems without manure input or antibiotic pollution.

Isotope Labeling to Study Phosphorus Uptake in the Arbuscular Mycorrhizal Symbiosis.

Isotope labeling enables the detection and quantification of nutrient fluxes between soil and plants through arbuscular mycorrhizal (AM) fungi. Here we describe the use of radioactive isotopes, 33P and 32P, to study the uptake of P from soil by AM fungal mycelium and its transfer to the host plant through the mycorrhizal pathway.

Effect of ash application on the decomposer food web and N mineralization in a Norway spruce plantation.

In the face of global climate change there is an increasing demand for biofuel, which exerts pressure on production and thus management of biofuel plantations. The intensification of whole-tree harvest from biofuel plantations increases export of nutrients. Returning ash from biofuel combustion to the forest plantations can amend the soil nutrient status and thus facilitate sustainable forest management. However, ash affects the forest floor decomposer food web, potentially changing organic matter turnover, carbon sequestration and nitrogen availability. Our aim was to examine the response of decomposer organisms, food web structure and nitrogen mineralization function after ash application. In a coniferous forest plantation amended with 0, 3, 4.5 or 6 t ash ha-1, we sampled in several depths of the forest floor for key organisms of the decomposer food web (fungal biomass, 0-12 cm; bacteria, protozoa, nematodes and enchytraeids, 0-3 cm and 3-6 cm; microarthropods and earthworms, 0-5 cm), 2, 14 and 26 months after ash application. We used structural equation modelling (SEM) to detangle the direct and indirect effects of ash application on organisms in the decomposer food web and on nitrogen availability. We found that ash increased the abundance of bacteria and protozoa, as well as the inorganic nitrogen pool at 0-3 cm depth, whereas the effect of ash was negligible at 3-6 cm depth. Earthworm abundance increased, whereas enchytraeid abundance decreased 2 years after ash application. The structural equation modelling showed that ash application stimulated the bacterial feeding pathway and increased nitrogen mineralization. Contrary, ash had a negative effect on fungal biomass at the first sampling, however, this effect subdued over time. Our results suggest that as the soil decomposer food web is resilient to ash application, this is a viable option for sustainable management of biofuel plantations.

Suppression of arbuscular mycorrhizal fungal activity in a diverse collection of non-cultivated soils.

Most plants form symbiotic associations with arbuscular mycorrhizal fungi (AMF). AMF increase the uptake of plant nutrients by extending their extra-radical mycelium (ERM) in the soil where other groups of microorganisms may suppress the activity of the ERM. However, little is known about such suppression in natural soils. This work aimed to investigate the incidence of AMF suppression among soils sampled from highly variable natural ecosystems, and used 33P uptake by the ERM to evaluate AMF activity. A second aim was to identify factors behind the observed AMF-suppression. We found that AMF-suppressiveness varied markedly among natural soils and occurred more frequently in low pH than in high pH soils. A previous study for cultivated soils revealed a strong biological component of suppressiveness against AMF, and in accordance we found that the composition of both fungal and bacterial communities differed significantly between AMF-suppressive and non-suppressive natural soils. Acidobacteria, Acidothermus, Xanthomonadaceae, Archaeorhizomyces sp., Mortierella humilis and some Mycena spp. were significantly more abundant in AMF-suppressive soils and may therefore be direct antagonists of AMF. This implies that the functioning of AMF in natural ecosystems is strongly modulated by specific soil microbes.

Suppression of the activity of arbuscular mycorrhizal fungi by the soil microbiota.

Arbuscular mycorrhizal fungi (AMF) colonise roots of most plants; their extra-radical mycelium (ERM) extends into the soil and acquires nutrients for the plant. The ERM coexists with soil microbial communities and it is unresolved whether these communities stimulate or suppress the ERM activity. This work studied the prevalence of suppressed ERM activity and identified main components behind the suppression. ERM activity was determined by quantifying ERM-mediated P uptake from radioisotope-labelled unsterile soil into plants, and compared to soil physicochemical characteristics and soil microbiome composition. ERM activity varied considerably and was greatly suppressed in 4 of 21 soils. Suppression was mitigated by soil pasteurisation and had a dominating biotic component. AMF-suppressive soils had high abundances of Acidobacteria, and other bacterial taxa being putative fungal antagonists. Suppression was also associated with low soil pH, but this effect was likely indirect, as the relative abundance of, e.g., Acidobacteria decreased after liming. Suppression could not be transferred by adding small amounts of suppressive soil to conducive soil, and thus appeared to involve the common action of several taxa. The presence of AMF antagonists resembles the phenomenon of disease-suppressive soils and implies that ecosystem services of AMF will depend strongly on the specific soil microbiome.

Risk assessment of replacing conventional P fertilizers with biomass ash: Residual effects on plant yield, nutrition, cadmium accumulation and mycorrhizal status.

Reutilizing biomass ashes in agriculture can substitute inputs of P from finite primary sources. However, recycling of ashes is disputed due to their content of toxic substances such as heavy metals. This study evaluates the potential risk of replacing easily soluble inorganic P fertilizer with P in biomass ashes in a barley crop grown on soil with adequate P status. Two contrasting doses of three different types of ashes were applied to an agricultural field with spring barley and compared to similar doses of triple-superphosphate fertilizer. In the second growing season after biomass ash application, grain, straw and root dry matter yield, and P and Cd uptake were determined. Resin-extractable P was measured in soil and the symbiotic arbuscular mycorrhizal fungal activity, colonization, and community composition were assessed. Crop yield was not affected by ash application, while P-uptake and mycorrhizal status were slightly enhanced with high ash applications. Changes to the mycorrhizal community composition were evident with high ash doses. Cadmium uptake in aboveground plant tissue was unaffected by ash treatments, but increased in roots with increasing doses. Consequently, we conclude that fertilization with biomass ashes can replace conventional fertilizers without risk to barley crops in the short term.

Antimicrobial, antiparasitic, anti-inflammatory, and cytotoxic activities of Lopezia racemosa.

The present study investigates the potential benefits of the Mexican medicinal plant Lopezia racemosa (Onagraceae). Extracts and fractions from aerial parts of this plant were assessed to determine their antibacterial, antifungal, antiparasitic, anti-inflammatory and cytotoxic activities in vitro. Aerial parts of the plant were extracted with various solvents and fractionated accordingly. Extracts and fractions were tested against a panel of nine bacterial and four fungal species. The antiparasitic activity was tested against Leishmania donovani, whereas the anti-inflammatory activity of the compounds was determined by measuring the secretion of interleukin-6 from human-derived macrophages. The same macrophage cell line was used to investigate the cytotoxicity of the compounds. Various extracts and fractions showed antibacterial, antifungal, antiparasitic, and anti-inflammatory activities. The hexanic fraction HF 11-14b was the most interesting fraction with antimicrobial, and anti-inflammatory activities. The benefit of L. racemosa as a traditional medicinal plant was confirmed as shown by its antibacterial, antifungal and anti-inflammatory activities. To the best of our knowledge, this is the first study reporting the biological activities of L. racemosa, including antiparasitic and anti-inflammatory activities.

Antimicrobial, anti-inflammatory, antiparasitic, and cytotoxic activities of Galium mexicanum.

ETHNOPHARMACOLOGICAL RELEVANCE: To study the potential benefit of the traditional Mexican medicinal plant Galium mexicanum Kunth (Rubiaceae). Hexane, chloroform, and methanol extracts as well as various fractions from these extracts were tested to determine antibacterial, antifungal, antiparasitic or anti-inflammatory activities in vitro. MATERIALS AND METHODS: Aerial parts of the plant were extracted with various solvents and fractionated accordingly. Their antibacterial and antifungal activities were assessed on nine bacterial and four fungal strains. Leishmania donovani was used as a protozoan strain for antiparasitic activity. The anti-inflammatory activity of the compounds was investigated by measuring the secretion of interleukin-6 when macrophages were exposed to lipopolysaccharide. RESULTS: Various extracts and fractions obtained from this plant exhibit antibacterial, antifungal, antiparasitic, and anti-inflammatory activities. Of special interest was the hexane fraction HE 14 b, which show antibacterial (ranging between 67 and 666 µg/ml) and antifungal (at concentrations of 333 µg/ml) activities. Also the hexane fraction HE 5 exhibited antiparasitic activity (at concentrations of 260 µg/ml), whereas the methanol fraction ME 13-15 showed a potent anti-inflammatory activity when compared to dexamethasone. Chemical analyses of the chloroform extract show the presence of triterpenes, saponins, flavonoids, sesquiterpene lactones, and glucosides, but no tannins were detected in the assayed extract. CONCLUSIONS: The benefit of Galium mexicanum as a traditional medicinal plant was confirmed using antibacterial and antifungal assays in vitro. We also report for the first time, and to the best of our knowledge, antiparasitic and anti-inflammatory activities of this plant.