• Energy Research

The species of the genus Trichoderma are used successfully as biocontrol agents against a wide range of phytopathogenic fungi. Among them, Trichoderma harzianum is especially effective. However, to develop more effective fungal biocontrol strategies in organic substrates and soil, tools for monitoring the control agents are required. Real-time PCR is potentially an effective tool for the quantification of fungi in environmental samples. The aim of this study consisted of the development and application of a real-time PCR-based method to the quantification of T. harzianum, and the extrapolation of these data to fungal biomass values. A set of primers and a TaqMan probe for the ITS region of the fungal genome were designed and tested, and amplification was correlated to biomass measurements obtained with optical microscopy and image analysis, of the hyphal length of the mycelium of the colony. A correlation of 0.76 between ITS copies and biomass was obtained. The extrapolation of the quantity of ITS copies, calculated based on real-time PCR data, into quantities of fungal biomass provides potentially a more accurate value of the quantity of soil fungi.

AbstractClimate change will affect semiarid ecosystems through severe droughts that increase the competition for resources in plant and microbial communities. In these habitats, adaptations to climate change may consist of thinning—that reduces competition for resources through a decrease in tree density and the promotion of plant survival. We deciphered the functional and phylogenetic responses of the microbial community to 60 years of drought induced by rainfall exclusion and how forest management affects its resistance to drought, in a semiarid forest ecosystem dominated byPinus halepensisMill. A multiOMICapproach was applied to reveal novel, community‐based strategies in the face of climate change. The diversity and the composition of the total and active soil microbiome were evaluated by 16SrRNAgene (bacteria) andITS(fungal) sequencing, and by metaproteomics. The microbial biomass was analyzed by phospholipid fatty acids (PLFAs), and the microbially mediated ecosystem multifunctionality was studied by the integration of soil enzyme activities related to the cycles of C, N, and P. The microbial biomass and ecosystem multifunctionality decreased in drought‐plots, as a consequence of the lower soil moisture and poorer plant development, but this decrease was more notable in unthinned plots. The structure and diversity of the total bacterial community was unaffected by drought at phylum and order level, but did so at genus level, and was influenced by seasonality. However, the total fungal community and the active microbial community were more sensitive to drought and were related to ecosystem multifunctionality. Thinning in plots without drought increased the active diversity while the total diversity was not affected. Thinning promoted the resistance of ecosystem multifunctionality to drought through changes in the active microbial community. The integration of total and active microbiome analyses avoids misinterpretations of the links between the soil microbial community and climate change.

Changing climatic conditions (warming and decreasing precipitation) have been found to be a threat to the agricultural sustainability of Mediterranean croplands. From the climate change perspective, biochar amendment may interact with the effects of warming and drought stresses on soil ecosystems. However, the responses of soil microbial communities to the joint effects of climate change and biochar in Mediterranean croplands are not sufficiently known. To help fill this knowledge gap, in this work we used a field experiment to determine the effects of partial rain exclusion alone or combined with a soil temperature increase in biochar-amended (20 t ha) and unamended plots under crop rotation on soil chemical properties, enzyme activities, and the microbial community activity, structure, composition, abundance, and functions. The biomass, composition, and activity of the soil bacterial and fungal communities were more responsive to biochar addition than to climate manipulation. Thus, soil chemical parameters, enzyme activities and the relative abundances of bacterial populations were not responsive to the interaction of biochar and climate manipulation, while the predicted functionality of the bacterial community was modified by both factors. Soil β-glucosidase activity significantly decreased in response to biochar addition and climate manipulation, while urease activity was significantly increased by biochar, and protease activity was significantly decreased by climate manipulation. Gram negative and fungal biomasses were significantly affected by the interaction of biochar with climate manipulation. Climate manipulation produced changes in the composition of the soil fungal community without loss of diversity. This study illustrates how the interactions between biochar amendment and future climate change scenarios influence microbially-driven ecosystem services related to the maintenance of nutrient cycles and biodiversity in a Mediterranean agroecosystem. This research was financially supported by the Spanish MICINN MINECO, AEI, FEDER, EU), through the research projects CGL2015-65162-R and AGL2016-75752-R. The authors are also grateful for the AEPP CSIC funds (2020AEP004). We also thank the Spanish Ministry and FEDER funds for the project AGL2017–85755-R (AEI/FEDER, UE), the i-LINK + 2018 (LINKA20069) from CSIC.

Forests influence climate and mitigate global change through the storage of carbon in soils. In turn, these complex ecosystems face important challenges, including increases in carbon dioxide, warming, drought and fire, pest outbreaks and nitrogen deposition. The response of forests to these changes is largely mediated by microorganisms, especially fungi and bacteria. The effects of global change differ among boreal, temperate and tropical forests. The future of forests depends mostly on the performance and balance of fungal symbiotic guilds, saprotrophic fungi and bacteria, and fungal plant pathogens. Drought severely weakens forest resilience, as it triggers adverse processes such as pathogen outbreaks and fires that impact the microbial and forest performance for carbon storage and nutrient turnover. Nitrogen deposition also substantially affects forest microbial processes, with a pronounced effect in the temperate zone. Considering plant-microorganism interactions would help predict the future of forests and identify management strategies to increase ecosystem stability and alleviate climate change effects. In this Review, we describe the impact of global change on the forest ecosystem and its microbiome across different climatic zones. We propose potential approaches to control the adverse effects of global change on forest stability, and present future research directions to understand the changes ahead.

Increased utilization of silver nanoparticles (AgNPs) can result in an accumulation of these particles in the environment. The potential detrimental effects of AgNPs in soil may be associated with the low fertility of soils in semiarid regions that are usually subjected to restoration through the application of organic amendments. Microbial communities are responsible for fundamental processes related to soil fertility, yet the potential impacts of low and realistic AgNPs concentrations on soil microorganisms are still unknown. We studied the effects of realistic citrate-stabilized AgNPs concentrations (0.015 and 1.5 μg kg-1) at two exposure times (7 and 30 days) on a sandy clay loam Mediterranean soil unamended (SU) and amended with compost (SA). We assessed soil microbial biomass (microbial fatty acids), soil enzyme activities (urease, β-glucosidase, and alkaline phosphatase), and composition of the microbial community (bacterial 16S rRNA gene and fungal ITS2 sequencing) in a microcosm experiment. In the SA, the two concentrations of AgNPs significantly decreased the bacterial biomass after 7 days of incubation. At 30 days of incubation, only a significant decrease in the Gram+ was observed at the highest AgNPs concentration. In contrast, in the SU, there was a significant increase in bacterial biomass after 30 days of incubation at the lowest AgNPs concentration. Overall, we found that fungal biomass was more resistant to AgNPs than bacterial biomass, in both SA and SU. Further, the AgNPs changed the composition of the soil bacterial community in SA, the relative abundance of some bacterial taxa in SA and SU, and fungal richness in SU at 30 days of incubation. However, AgNPs did not affect the activity of extracellular enzymes. This study demonstrates that the exposure time and organic amendments modulate the effects of realistic concentrations of AgNPs in the biomass and composition of the microbial community of a Mediterranean soil.

The increasing use of silver nanoparticles (AgNPs) due to their well-known antimicrobial activity, has led to their accumulation in soil ecosystems. However, the impact of environmental realistic concentrations of AgNPs on the soil microbial community has been scarcely studied. In this work, we have assessed the impact of AgNPs, that mimic real concentrations in nature, on tropical soils cultivated with Coffea arabica under conventional and organic management systems. We evaluated the biomass, extracellular enzyme activities, and diversity of the soil microbial community, in a microcosm experiment as a function of time. After seven days of incubation, we found an increase in microbial biomass in an AgNPs-concentration-independent manner. In contrast, after 60-day-incubation, there was a decrease in Gram+ and actinobacterial biomass, in both soils and all AgNPs concentrations. Soil physico-chemical properties and enzyme activities were not affected overall by AgNPs. Regarding the microbial community composition, only some differences in the relative abundance at phylum and genus level in the fungal community were observed. Our results suggest that environmental concentrations of AgNPs affected microbial biomass but had little impact on microbial diversity and may have little effects on the soil biogeochemical cycles mediated by extracellular enzyme activities.

Lignocellulose from plant biomass represents an abundant and renewable source for the production of environmentally friendly chemicals and biofuels. However, its recalcitrant nature entails the use of complex biochemical reactions that are still challenging. Since the degradation of lignocellulose is the current bottleneck of the conversion processes, the search for novel enzymes and microbial strains for degrading plant biomass is of high importance. Currently, bacteria are in the spotlight as promising candidates for novel conversion strategies due to their wide functional diversity and versatility. Here, we review the lines of evidence that show the high potential of bacterial strains from soil for biomass conversion ranging from strain characterization to metagenome and metatranscriptome analysis. Substantial and diverse fractions of soil bacteria are able to decompose the major lignocellulose components. To do that, bacteria evolved structurally variable and often highly complex lignocellulolytic systems composed of enzymes as well as proteins involved in efficient substrate binding. Both as individual components or in combination, bacterial enzymes, and accessory proteins appear to be promising tools in the biotechnological valorization of lignocellulose.

The growth and survival of plants in semiarid Mediterranean forests can be improved through the benefits conferred by thinning, a forest management practice that removes trees and reduces the competition between the remaining ones. Here, we evaluate the impacts of induced drought (the exclusion of 25% of the natural rainfall for 5 years) and thinning, and their interaction, with the objective of determining whether the thinning of Holm oak (Quercus ilex L.) modulates the resistance of the soil microbial community to drought. Sequencing of 16S rRNA and ITS amplicons revealed that drought, thinning, and their interaction influenced the composition of the bacterial community, while the fungal community was exclusively affected by thinning. Thinning consisted of the removal of the aboveground parts of the Holm oak trees, which were thereafter left in forest stand. Thinning contributed to the C and N contents, with parallel increases in microbial biomass, particularly in summer. Drought increased the amounts of total organic C and total N, likely due to the reduced enzyme activities. Indeed, the composition of the bacterial community was modulated primarily by the indirect and long-term effects of drought - the accumulation of soil organic matter - rather than by the direct effect of the lower water content imposed by the drought treatments. Thinning under drought conditions did not increase soil organic C (SOC) content. However, the resistance of the soil microbial community to drought was fostered by thinning, particularly at the functional level, as indicated by the enzyme activities related to C, N and P cycles. These responses were associated to variations in the composition of the microbial communities in thinned, drought-exposed plots, in comparison to unthinned, drought-exposed plots. In conclusion, the interaction between forest management and drought influenced the soil microbial community of a Holm oak-dominated Mediterranean ecosystem.