The responses of Spartina alterniflora Loisel. roots to the interactive effects of drought and nitrogen (N) form, and the underlying mechanisms involved, remain poorly understood. We conducted a greenhouse experiment to evaluate the effects of N form (NH4 +, NO3 -, and NO3 -/NH4 +) and increasing water deficit on root performance, including growth, metabolite profiles, antioxidant activity, and N metabolism. Under well-watered conditions, NH4 +-fed plants exhibited the greatest root growth, nearly double that of NO3 --fed plants. However, this growth advantage was lost under mild (50% field capacity, FC) and severe (25% FC) drought stress. In contrast, drought stress enhanced root growth in NO3 --fed plants relative to well-watered conditions. Under well-watered conditions, NH4 + nutrition increased the activities of superoxide dismutase, glutathione reductase, and ascorbate peroxidase compared to NO3 - nutrition. Although drought stress further stimulated antioxidant enzyme activities in the roots of NH4 +-fed plants, this response did not mitigate drought-induced growth reductions. Antioxidant enzyme activities in the NO3 -- and NO3 -/NH4 +-fed plants were largely unaffected by drought, except for guaiacol peroxidase. Regardless of N form, glutamine synthetase activity increased under mild drought stress but declined under severe stress. Drought stress also enhanced glutamate dehydrogenase activity across all N treatments, particularly in NH4 +-fed plants, and was accompanied by increased total amino acid concentrations, especially proline. Despite these metabolic adjustments, drought stress reduced the overall performance of NH4 +-fed plants. These findings provide insights into N form-dependent drought responses and may help guide fertilizer management strategies to improve S. alterniflora productivity under water-limited conditions.

Previously published data showed the ubiquity of thermophilic bacteria in upper soil layers and their potential significant role in biogeochemical cycles. The processes for the maintenance of cell viability by these thermophiles in soils, including cool temperate soils, are largely unknown. We used culturing systems to mimic and analyze usual environmental growth-limiting conditions and near-zero growth rates, namely those imposed by carbon availability, and common in soils. Our goal was to comprehend how a thermophilic bacterium of the Bacillota Phylum, Parageobacillus thermoglucosidasius 23.6, persists and maintains its viability in upper soils. Comparative transcriptomic analysis of P. thermoglucosidasius 23.6 at optimum growth rate (2.2 h−1), slow growth (0.025 h−1 and 0.002 h−1) and near-zero growth rate (0.0002 h−1) revealed the overexpression of [NiFe]-hydrogenase-encoding genes, specifically of those encoding a putative Huc-type high affinity [NiFe]-hydrogenase, under growth limiting conditions. High affinity [NiFe]-hydrogenases were previously shown to be enzymes yielding energy during carbon starvation and to have a major role in the oxidation of tropospheric H2 in soil ecosystems; their activity has been proposed as a major sink for global atmospheric H2. The presence and expression of these high affinity [NiFe]-hydrogenase-encoding genes are suggested to represent a widespread strategy of terrestrial bacteria, specifically of soil thermophiles, to stay energized among resource variability or limitation, which could be considered a critical mechanism to maintain viability under growth limiting conditions to ensure long-term persistence in soils.

Rural areas face increasing pressures from demographic decline, climate change, and the transition towards sustainable food systems. This work examines how Youth Entrepreneurship (YE), Smart Farming (SF), and Living Labs (LLs) can be integrated to revitalise rural development and agricultural transition. Drawing on European Union (EU) policy frameworks, academic literature, and insights from recent research projects, we develop a conceptual model that highlights the synergies and gaps between these three domains. We argue that SF provides technological opportunities for productivity and sustainability, while LLs act as innovation ecosystems that enable co-creation and participatory governance. Together, they can foster YE by enhancing skills, reducing uncertainty, and embedding young farmers in collaborative networks. However, significant challenges remain, including the limited adaptation of SF to diverse farming contexts, the uneven capacity of LLs to sustain innovation partnerships, and insufficient policy measures to attract and retain youth in rural areas. By synthesising technological, social, and entrepreneurial perspectives, this study contributes to the foresight debate on how digital transitions can support rural resilience and long-term sustainability. The findings are relevant to researchers, policymakers, and practitioners seeking to strengthen the role of youth in shaping the future of agriculture and rural societies.

Biostimulants represent more than an agricultural product; they constitute a paradigm shift towards more sustainable food production systems. This editorial examines how biostimulants promote a transition from conventional agriculture, dependent on synthetic inputs, to environmentally conscious and regenerative agricultural practices. Historically, conventional agriculture has relied on synthetic fertilizers, pesticides, and monocultures to maximize production, resulting in significant environmental costs such as soil degradation, water pollution, and loss of biodiversity. Biostimulants offer an alternative by improving soil health through the support of microbiomes, increasing plant tolerance to abiotic stress, and reducing dependence on chemical inputs. According to the European Union, biostimulants are products that stimulate plant nutritional processes, improving nutrient use efficiency, tolerance to abiotic stress, quality traits, or the availability of poorly soluble nutrients. The global biostimulant market is expected to grow from $3.2 billion in 2023 to over $9.0 billion in 2030. The BioClub project demonstrated that biofertilizers composed of microbial consortia can compensate for a 33% reduction in the recommended dose of chemical fertilizers, maintaining productivity and enriching grains with zinc and iron. The effective implementation of biostimulants requires multi-actor approaches, such as living labs, integrating researchers, producers, technicians, and policymakers to optimize their application in different agroecosystems.

In the realm of sustainable and eco-friendly agriculture, current scientific research emphasizes the development of plant-based bioproducts to mitigate the agricultural footprint resulting from excessive fertilizer and pesticide use. This study investigates the cladodes of Opuntia ficus-indica to screen for bioactive compounds and assess their efficacy against fungal pathogens isolated from infected tomato fruits. Quantitative analysis of the methanolic extract revealed substantial concentrations of bioactive compounds: total polyphenols (86.6 mg GAEs/100 g FW), flavonoids (13.4 mg QEs/100 g FW), condensed tannins (8.9 mg TAEs/100 g FW), and carotenoids (0.9 mg β-CEs/100 g FW). Notably, the DPPH assay indicated that the cladode extract exhibited significant antioxidant potential at a concentration of 0.6 mg/mL. Seven fungal pathogens were isolated from infected tomato fruits and identified as belonging to the following genera: Rhizoctonia (EC2), Fusarium (EC1 and EC3), Alternaria (EC4), Mucor (EC5), Aspergillus (EC6), and Penicillium (EC7). At a concentration of 0.02% of the cladode hydroethanolic extract, the antifungal activity results demonstrated mycelial growth inhibition for Alternaria sp. (70.91%), Rhizoctonia solani EC2 (58.49%), Fusarium oxysporum EC3 (57.63%), and Fusarium solani EC1 (53.13%). Conversely, lower inhibitory activities were observed for Mucor sp. EC5 (31.08%), Aspergillus sp. EC6 (35.14%), and Penicillium sp. EC7 (28.38%). At a concentration of 0.04%, all cladode hydroethanolic extracts inhibited mycelial growth by more than 50%. Furthermore, the highest spore inhibition was attained with the 0.04% cladode hydroethanolic extract (exceeding 50%). Inhibition percentages of 83.02%, 85.96%, 87.76%, and 90.20% were recorded for Fusarium oxysporum EC3, Fusarium solani EC1, Rhizoctonia solani EC2, and Alternaria sp. EC4, respectively. Collectively, these findings suggest that Opuntia ficus-indica extract holds significant promise for application as a biopesticide against fungal pathogens affecting tomato fruits.

Dry grasslands are vast, socioeconomically and ecologically important environments, which are increasingly threatened by multiple stressors. We tested whether plant cover composition could mitigate ecosystem services loss under multiple stressors in dry grassland mesocosms by growing the grass sorghum (Sorghum bicolor) alone (Grass cover) or together with the legume serradella (Ornithopus sativus) (Mixed cover) under frequent cutting and/or increasing water stress. We assessed erosion control, carbon sequestration, forage quantity and quality, and soil fertility, individually and simultaneously (i.e., multifunctionality). Contrary to our hypothesis, the Mixed cover did not improve ecosystem services compared to the Grass cover, except for forage quality, which improved by 30%. In general, the stressors had negative effects: cutting reduced erosion control by 20%, forage quantity by 50%, soil fertility by 40% and multifunctionality by 20%, and severe water stress decreased carbon sequestration by 40%, forage quantity by 30%, soil fertility by 10%, and multifunctionality by 10%. Water stress caused 100% serradella mortality, underscoring this legume’s vulnerability to increasing aridity. Combined stressors yielded the lowest service provision. Forage quality was the only service that improved under stress: cutting improved it by 40% and severe water stress by 60%. Our results suggest that while systems combining grasses and legumes may enhance forage quality, grass-dominated systems appear more resilient to multiple stressors in drylands, largely due to their superior efficiency in accessing and conserving limited water and nutrient resources. Given the ongoing trends of aridification and land-use intensification, future research should explore adaptive management strategies that prioritize resource-efficient plant species, foster belowground resource retention, and optimize grazing regimes to sustain resilience and multifunctionality in dry grasslands.

Forest soils represent bustling metropolises of invisible microbial life, where bacteria, fungi, archaea, algae, and protozoa create complex networks essential for ecosystem functioning. This opinion paper examines the critical role of soil biomes in forest productivity and sustainability, comparing microbial communities in forest versus agricultural environments. Forest soils, dominated by mycorrhizal fungi, maintain high microbial diversity and stable networks that support nutrient cycling, carbon sequestration, and plant resilience. In contrast, agricultural soils often exhibit reduced microbial diversity due to human interventions such as monocropping, pesticide use, and chemical fertilization. The rhizosphere emerges as a hotspot of microbial activity, where root secretions and associated microorganisms form interactive networks crucial for biogeochemical processes. Key microbial players include ectomycorrhizal and arbuscular mycorrhizal fungi, which enhance nutrient uptake and plant stress tolerance. A biome-based strategy for maintaining forest soil health emphasizes reducing mechanical disturbances, promoting plant biodiversity, and incorporating organic amendments like biochar and compost. Successful examples worldwide demonstrate the effectiveness of microbial inoculation, from ectomycorrhizal fungi in pine and oak forestry to nitrogen-fixing bacteria in eucalyptus plantations. Adaptive management approaches utilizing metagenomics and soil health indicators enable informed decision-making for long-term ecosystem stability. Understanding forest soil biomes is essential for effective forest management, biodiversity preservation, and sustainable resource use, particularly as climate change intensifies the need for resilient forest ecosystems.

This study develops sustainable, antibacterial food packaging films using carboxymethylcellulose and fungi- derived chitin nanofibrils (ChNFs) reinforced with clay to enhance mechanical strength, moisture resistance, and gas barrier properties. ChNFs significantly improve tensile strength and permeability by forming a dense, hydrogen-bonded network within the carboxymethylcellulose matrix. However, excessive ChNF content led to agglomeration, reducing mechanical performance slightly. At 30% ChNFs content, films demonstrated antibacterial activity against Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes and also presented a 52.1 +/- 3.2% degradation rate in four weeks. Life cycle assessment revealed a reduced carbon footprint (5.0-5.3 kg COQ-equiv. per kg film) and low plastic litter generation (35-44 g/kg), underscoring environmental benefits compared to conventional packaging. These carboxymethylcellulose/ChNF-based films are a promising, ecofriendly alternative for food packaging applications, offering antibacterial properties and enhanced sustain- ability in the packaging of perishable food products.

Ensuring food security in a framework of environmental sustainability is the greatest challenge of the 21st century. The rapid population growth together with changing consumption patterns associated with new lifestyles mean that total demand for food is increasing at a faster pace than that of the production capacity. To overcome this challenge we need to transform agriculture into a more efficient and less polluting activity. This may be achieved through harnessing the trillions of organisms inhabiting the soil. In this work we show how the use of microbial consortia can contribute to increased productivity and efficient use of nutrients in a maize field. The combined use of arbuscular mycorrhizal fungi and bacteria that promote plant growth, when applied in a field experiment were able to compensate for the reduction in fertilizer by 33%. The results show that the application of the microbial consortium increased nitrogen fixing and phosphorus solubilising bacteria in the soil, which may explain the increased uptake of these nutrients by the plants.

Avian colibacillosis remains a major threat to poultry production and food security, whereas its antibiotic-based control accelerates antimicrobial resistance. This study investigated the antibacterial potential of the hydroethanolic extract of Thymus vulgaris, alone and in combination with antibiotics, against avian pathogenic Escherichia coli (APEC). The phytochemical composition of the extract was analyzed by HPLC, while its antibacterial activity was assessed using agar diffusion and minimum inhibitory concentration (MIC) assays. Antibiotic-extract interactions were evaluated by the checkerboard method, and mechanisms of action were explored through assays targeting membrane permeability, proton pump inhibition, catalase inhibition, oxidative stress, and biofilm formation. Its antioxidant, anti-inflammatory, and cytotoxic activities were also evaluated. HPLC analysis revealed 16 phenolic compounds, including rosmarinic, ferulic, and salicylic acids, as well as flavonoids such as quercetin and luteolin, with an extraction yield of 15% ± 0.55. The extract displayed significant antibacterial activity (MIC: 5.46-10.93 mg/ml, p < 0.05). In synergy tests, the extract enhanced ampicillin efficacy 4-8-fold and showed additive effects with ciprofloxacin and tetracycline (2-4-fold). Mechanistic assays demonstrated disruption of bacterial membranes (32.66% electrolyte leakage, p < 0.05), inhibition of H⁺-ATPase proton pumps (p < 0.05), catalase activity reduction by 79.2% (p < 0.05), >50% decrease in cell hydrophobicity, and inhibition of biofilm formation (49.07-72.47%), alongside eradication of mature biofilms (41.89-64.67%) (p < 0.05). Beyond antimicrobial effects, the extract exhibited notable antioxidant capacity. In DPPH radical scavenging, the extract showed an IC₅₀ of 9.76 ± 0.228 μg/ml (p < 0.05), while in the TAC assay, it reached 467.25 ± 1.889 μg/ml (p < 0.05), and it reduced ampicillin-induced lipid peroxidation by 42.85% (p < 0.05). Anti-inflammatory testing revealed inhibition of protein denaturation (4.95-52.48% at 15-2000 µg/ml, p < 0.05), although weaker than aspirin. Hemolysis assays confirmed the extract was non-hemolytic at concentrations up to 11 mg/ml (p < 0.05). In conclusion, Thymus vulgaris extract demonstrated safe, multi-target bioactivity, supporting its potential as a promising alternative to combat avian colibacillosis and antimicrobial resistance.

Inoculation with the PGPB Herbaspirillum seropedicae shapes both the structure and putative functions of the wheat microbiome and causes changes in the levels of various plant metabolites described to be involved in plant growth and health. Plant growth promoting bacteria (PGPB) can establish metabolic imprints in their hosts, contributing to the improvement of plant health in different ways. However, while PGPB imprints on plant metabolism have been extensively characterized, much less is known regarding those affecting plant indigenous microbiomes, and hence it remains unknown whether both processes occur simultaneously. In this study, both 16S amplicon and ITS sequencing analyses were carried out to study both the structural as well as the putative functional changes in the seed-borne endophytic microbiome of wheat plants inoculated with the PGPB Herbaspirillum seropedicae strain RAM10. Concomitantly, Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) analyses were used to investigate the alterations in the root metabolome of PGPB-inoculated plants. PGPB inoculation led to marked differences in the composition of the root microbiome, accompanied by the differential enrichment of microorganisms with putative roles in both plant energy and nitrogen metabolism. In addition, metabolome analyses showed that the levels of 16 metabolites belonging to the phenylpropanoid, terpenoid, and unsaturated fatty acid families were significantly altered in PGPB-inoculated plants. These findings shed light on the interplay between PGPB, the plant and its associated microbiome, indicating that PGPB can act as the driving force mediating long-lasting changes in both the plant metabolome and the plant microbiome.

Carthamus caeruleus L. is traditionally used in Algerian medicine, particularly for burn treatment, but its therapeutic potential remains insufficiently studied. This study aimed to evaluate the antioxidant and anti-inflammatory properties of the root aqueous extract, and to perform phytochemical characterization to identify its bioactive compounds. Phytochemical analysis was conducted using spectrophotometry and reverse-phase high-performance liquid chromatography (RP-HPLC). The antioxidant potential was assessed through various assays, including ferric reducing antioxidant power (FRAP), total antioxidant capacity (TAC), DPPH radical scavenging, hydroxyl radical scavenging, ferrous ion chelation, and hydrogen peroxide decomposition. Anti-inflammatory activity was evaluated using membrane stabilization, protein denaturation, and membrane peroxidation assays. The extract exhibited moderate levels of polyphenols, flavonoids, and condensed tannins, quantified as 21.19 ± 0.37 mg GAE/g, 0.72 ± 0.013 mg QE/g, and 27.28 ± 1.04 mg TAE/g of dry extract, respectively. RP-HPLC analysis identified 22 phytochemical compounds, primarily phenolic acids, flavonoids, and tannins, with orientin and vanillin as the major constituents. The extract demonstrated significant antioxidant activity, with moderate efficacy in TAC and FRAP assays (IC50 values of 5405.1 ± 4.42 and 1132.35 ± 4.97 µg/mL, respectively). Notable activities included DPPH and hydroxyl radical scavenging (34.43 ± 4.83 and 512.81 ± 9.46 µg/mL, respectively), ferrous ion chelation (2462.76 ± 1.38 µg/mL), lipid peroxidation inhibition (22.32 ± 3.31%), and hydrogen peroxide decomposition (263.93 ± 7.87 µg/mL). Additionally, the extract stabilized erythrocyte membranes under osmotic, thermal, and oxidative stress conditions (98.13 ± 0.15%, 70 ± 1.27%, and 89 ± 0.87%, respectively), inhibited ovalbumin denaturation (81.05 ± 2.2%), and protected against lipid peroxidation in brain homogenates (69.25 ± 0.89%). These findings support the traditional therapeutic applications of C. caeruleus and highlight its potential as a source of antioxidant and anti-inflammatory agents.

This study aimed to isolate glyphosate-tolerant bacteria and characterize their plant-beneficial traits. Using a bioprospecting and glyphosate enrichment methodology, we isolated 35 bacterial strains that were tested for tolerance to glyphosate on solid and liquid media (0, 11, 22, 44, and 88 g L-1 of glyphosate). Sixteen isolates with high tolerance to glyphosate (>= 22 g L-1) were identified and characterized by their ability to fix atmospheric nitrogen, solubilize inorganic phosphate, and produce indole-3-acetic acid (IAA). After similarity analysis and compatibility testing, a consortium composed of three bacteria (Priestia sp.; Achromobacter sp.; Pseudomonas sp.) were selected to be applied in a model plant (tomato) and Arabica coffee beans. The bacterial consortium promoted the germination (96.6%) and development of tomato seedlings, while also degrading residual glyphosate (31.25%) in coffee beans, as evidenced by the reduced levels of the herbicide. These results demonstrate the bacterial consortium's potential to reduce glyphosate residues and sustainably increase plant productivity.

Previous studies have highlighted the widespread presence of thermophilic bacterial genera in upper soil layers, their role in biogeochemical cycles, and their potential application in soil fertilization. However, the mechanisms by which these thermophiles maintain cell viability in temperate soils remain largely unknown. The isolation of thermophilic bacteria from rhizospheric soils has been reported, hence it may be hypothesized that the rhizosphere environment plays a role in their survival. In this study, we developed a hydroponic system to introduce the thermophilic bacterium Parageobacillus thermoglucosidasius into the rhizosphere of tomato plants, demonstrating that this environment increased bacterial survival rates at 20°C-25°C by over 23-fold. The rhizosphere exudates contributed to this increase, as their addition boosted bacterial survival in pure cultures at 25°C by up to twofold. We propose that the rhizosphere and its exudates, characterized through targeted metabolomics, support the persistence of thermophilic bacteria in temperate soils during colder periods, ensuring viable cells that contribute to soil fertilization during warmer seasons.

Soil aggregate stability has been asserted as an indicator of soil quality. Earthworms are essential for the stability of soil aggregates due to their activities, including burrowing and organic matter decomposition. In this study, we focused on how differences in plant litter quality influences earthworm abundance and composition and, subsequently, macroaggregate stability in an agroforestry ecosystem. Litter from two dominant plant species (Quercus suber L. and Agrostis pourretii Willd.) were collected in an eco-intensively managed agroforestry ecosystem (eco-intensive farm) and in a neighbor conventionally managed agroforest (conventional farm). Then, a field experiment was implemented in the eco-intensive farm, composed of 6 litter treatments (control with no litter; Q. suber senescent leaves from the eco-intensive farm; A. pourretii tussocks from the eco-intensive farm; a mixture of the previous two; Q. suber senescent leaves from the conventional farm; A. pourretii tussocks from the conventional farm), replicated across 3 areas of the farm, with two blocks per area (a total of 6 replicates per treatment). Litter decomposition rates for the different treatments were quantified over 15 months. At the end of the experiment, soil from each treatment was sampled to measure macroaggregate stability and other soil parameters (i.e., moisture, organic matter, pH, TC, TOC, P, N, NO3- , NH4+, humic and fulvic acids, water repellence), as well as earthworms. Earthworm abundance had a positive effect on macroaggregate stability (as expressed by the ratio between stable macroaggregates and the total amount of soil), irrespective of litter quality, while litter quality had no direct effect on macroaggregate stability. According to our study's findings, earthworms have a significant intervenient role in the stabilization of soil aggregates in Mediterranean agroforests. This emphasizes the necessity for soil management practices that maintain earthworm communities and their beneficial contributions to soil health.

When aiming to increase plants' nitrogen (N) budget, special attention is given to the microbial inoculum's capacity to perform biological N₂ fixation. However, we consider that other approaches can be explored. Here, we report initial results of plant growth promoting rhizobacteria (Azospirillum brasilense strains Sp245 and ARG2) capacity to scavenge atmospheric ammonia (NH₃). Using a bipartite Petri dish system, we grew the two A. brasilense strains with the appropriate controls, and with atmospheric NH₃ as a N source. By increasing the atmospheric NH₃ concentration, the growth rate of both A. brasilense strains increased almost 4 times in relation to the controls. By creating a gradient of atmospheric NH₃ concentrations we changed the growth rate of both A. brasilense strains, but its effect differed between the two bacterial strains, i.e., the Sp245 strain increased its growth rates up to pH 9.0, while the ARG2 strain reached maximum growth rates at pH 9.5. The fact that these two plant growth promoting rhizobacteria scavenge atmospheric NH₃, instead of fixing N₂, suggests that this overlooked microbial trait can be an interesting tool to mitigate atmospheric NH₃ concentrations, especially in farming environments.

Both, Serendipita indica and AMF, show promise as sustainable biofertilizers for reforestation, improving nutrient uptake and stress tolerance, despite contrasting effects on photosynthetic capacity and biomass allocation. Reclaiming degraded areas is essential for biodiversity conservation and enhancing ecosystem services enhancement, especially when using native species. This study investigated Schinus terebinthifolius Raddi, a native Brazilian species, and its compatibility with plant growth-promoting microorganisms (PGPM), including an endophytic fungus (Serendipita indica) and a consortium of arbuscular mycorrhizal fungi (AMF), to identify effective strategies for reforestation in nutrient-poor environments. We observed growth stimulation by both PGPMs; however, S. indica primarily enhanced root weight, whereas AMF improved shoot weight. S. indica’s positive effects on root systems could be attributed to increased auxin levels and altered root architecture, which are critical for seedling establishment in reforestation programs. In terms of nutritional status, both treatments increased the content of most nutrients, with higher micronutrient contents in the shoots and higher macronutrient content in roots of inoculated plants. Despite AMF’s role in enhancing photosynthesis, plants inoculated with these fungi showed reduced photosynthetic capacity traits, possibly due to lower leaf nitrogen content. The proteomic analysis of Schinus terebinthifolius leaf extracts revealed that, despite the upregulation of several proteins associated with the photosynthetic apparatus in response to S. indica treatment, no enhancement in photosynthetic capacity was observed. We also found several proteins related to oxidative stress in plants inoculated with both fungi, indicating a greater tolerance to adverse environmental conditions. These findings underscore the potential of both, S. indica and AMF, as sustainable alternatives to chemical fertilizers in reforestation efforts, enhancing seedling quality and survival in nutrient-poor soils.

Biostimulants towards sustainable food production Cristina Cruz and Teresa Dias from Faculdade de Ciências da Universidade de Lisboa, focus on biostimulants, which are more than a product, but a significant change towards sustainable food production systems.