A Metagenomic and Gene Expression Analysis in Wheat (T. durum) and Maize (Z. mays) Biofertilized with PGPM and Biochar.

Commodity crops, such as wheat and maize, are extremely dependent on chemical fertilizers, a practice contributing greatly to the increase in the contaminants in soil and water. Promising solutions are biofertilizers, i.e., microbial biostimulants that when supplemented with soil stimulate plant growth and production. Moreover, the biofertilizers can be fortified when (i) provided as multifunctional consortia and (ii) combined with biochar with a high cargo capacity. The aim of this work was to determine the molecular effects on the soil microbiome of different biofertilizers and delivery systems, highlight their physiological effects and merge the data with statistical analyses. The measurements of the physiological parameters (i.e., shoot and root biomass), transcriptomic response of genes involved in essential pathways, and characterization of the rhizosphere population were analyzed. The results demonstrated that wheat and maize supplemented with different combinations of selected microbial consortia and biochar have a positive effect on plant growth in terms of shoot and root biomass; the treatments also had a beneficial influence on the biodiversity of the indigenous rhizo-microbial community, reinforcing the connection between microbes and plants without further spreading contaminants. There was also evidence at the transcriptional level of crosstalk between microbiota and plants.

Molecular and genetic bases of heat stress responses in crop plants and breeding for increased resilience and productivity.

To ensure the food security of future generations and to address the challenge of the 'no hunger zone' proposed by the FAO (Food and Agriculture Organization), crop production must be doubled by 2050, but environmental stresses are counteracting this goal. Heat stress in particular is affecting agricultural crops more frequently and more severely. Since the discovery of the physiological, molecular, and genetic bases of heat stress responses, cultivated plants have become the subject of intense research on how they may avoid or tolerate heat stress by either using natural genetic variation or creating new variation with DNA technologies, mutational breeding, or genome editing. This review reports current understanding of the genetic and molecular bases of heat stress in crops together with recent approaches to creating heat-tolerant varieties. Research is close to a breakthrough of global relevance, breeding plants fitter to face the biggest challenge of our time.

Marine Bioactive Peptides-An Overview of Generation, Structure and Application with a Focus on Food Sources.

The biggest obstacles in the application of marine peptides are two-fold, as in the case of non-marine plant and animal-derived bioactive peptides: elucidating correlation between the peptide structure and its effect and demonstrating its stability in vivo. The structures of marine bioactive peptides are highly variable and complex and dependent on the sources from which they are isolated. They can be cyclical, in the form of depsipeptides, and often contain secondary structures. Because of steric factors, marine-derived peptides can be resistant to proteolysis by gastrointestinal proteases, which presents an advantage over other peptide sources. Because of heterogeneity, amino acid sequences as well as preferred mechanisms of peptides showing specific bioactivities differ compared to their animal-derived counterparts. This review offers insights on the extreme diversity of bioactivities, effects, and structural features, analyzing 253 peptides, mainly from marine food sources. Similar to peptides in food of non-marine animal origin, a significant percentage (52.7%) of the examined sequences contain one or more proline residues, implying that proline might play a significant role in the stability of bioactive peptides. Additional problems with analyzing marine-derived bioactive peptides include their accessibility, extraction, and purification; this review considers the challenges and proposes possible solutions.

Meta-Analysis for Correlating Structure of Bioactive Peptides in Foods of Animal Origin with Regard to Effect and Stability.

Amino acid (AA) sequences of 807 bioactive peptides from foods of animal origin were examined in order to correlate peptide structure with activity (antihypertensive, antioxidative, immunomodulatory, antimicrobial, hypolipidemic, antithrombotic, and opioid) and stability in vivo. Food sources, such as milk, meat, eggs, and marine products, show different frequencies of bioactive peptides exhibiting specific effects. There is a correlation of peptide structure and effect, depending on type and position of AA. Opioid peptides contain a high percentage of aromatic AA residues, while antimicrobial peptides show an excess of positively charged AAs. AA residue position is significant, with those in the first and penultimate positions having the biggest effects on peptide activity. Peptides that have activity in vivo contain a high percentage (67%) of proline residues, but the positions of proline in the sequence depend on the length of the peptide. We also discuss the influence of processing on activity of these peptides, as well as methods for predicting release from the source protein and activity of peptides.

Plant Response to Metal-Containing Engineered Nanomaterials: An Omics-Based Perspective.

The increasing use of engineered nanomaterials (ENMs) raises questions regarding their environmental impact. Improving the level of understanding of the genetic and molecular basis of the response to ENM exposure in biota is necessary to accurately assess the true risk to sensitive receptors. The aim of this Review is to compare the plant response to several metal-based ENMs widely used, such as quantum dots, metal oxides, and silver nanoparticles (NPs), integrating available "omics" data (transcriptomics, miRNAs, and proteomics). Although there is evidence that ENMs can release their metal components into the environment, the mechanistic basis of both ENM toxicity and tolerance is often distinct from that of metal ions and bulk materials. We show that the mechanisms of plant defense against ENM stress include the modification of root architecture, involvement of specific phytohormone signaling pathways, and activation of antioxidant mechanisms. A critical meta-analysis allowed us to identify relevant genes, miRNAs, and proteins involved in the response to ENMs and will further allow a mechanistic understanding of plant-ENM interactions.

Molecular Response of Crop Plants to Engineered Nanomaterials.

Functional toxicology has enabled the identification of genes involved in conferring tolerance and sensitivity to engineered nanomaterial (ENM) exposure in the model plant Arabidopsis thaliana (L.) Heynh. Several genes were found to be involved in metabolic functions, stress response, transport, protein synthesis, and DNA repair. Consequently, analysis of physiological parameters, metal content (through ICP-MS quantification), and gene expression (by RT-qPCR) of A. thaliana orthologue genes were performed across different plant species of agronomic interest to highlight putative biomarkers of exposure and effect related to ENMs. This approach led to the identification of molecular markers in Solanum lycopersicum L. and Cucurbita pepo L. (tomato and zucchini) that might not only indicate exposure to ENMs (CuO, CeO2, and La2O3) but also provide mechanistic insight into response to these materials. Through Gene Ontology (GO) analysis, the target genes were mapped in complex interatomic networks representing molecular pathways, cellular components, and biological processes involved in ENM response. The transcriptional response of 38 (out of 204) candidate genes studied varied according to particle type, size, and plant species. Importantly, some of the genes studied showed potential as biomarkers of ENM exposure and effect and may be useful for risk assessment in foods and in the environment.

Special Issue Physiological and Molecular Responses of Plants to Engineered Nanomaterials.

Engineered nanomaterials (ENMs), by definition materials with a size between 1 and 100 nm, are becoming an important part of the economy and thanks to their many fields of applications, from photovoltaic cells to fertilizers, are increasingly coming into contact with plants and with the environment in general [...].

Long-term exposure to ambient air pollution and inflammatory response in the KORA study.

Previous studies consistently showed an association between fine atmospheric particulate matter (PM2.5) and cardiovascular diseases. Concerns about adverse health effects of ultrafine particles (UFP) are growing but long-term studies are still scarce. In this study, we examined the association between long-term exposure to ambient air pollutants and blood biomarkers of inflammation and coagulation, including fibrinogen, high-sensitivity C-reactive protein (hs-CRP), serum amyloid A (SAA) adiponectin and interleukin-6 (IL-6), measured in the German KORA-S4 cohort study (1999-2001). IL-6 was available for older participants only, who were therefore considered as a subsample. Annual mean concentrations of UFP (as particle number concentration), particulate matter in different particles sizes (PM10, PMcoarse, PM2.5, PM2.5 absorbance), ozone (O3), and nitrogen oxides (NO2, NOX) were estimated by land-use regression models and assigned to participants' home addresses. We performed a multiple linear regression between each pollutant and each biomarker with adjustment for confounders. Per 1 interquartile range (IQR, 1945 particles/cm3) increase of UFP, fibrinogen increased by 0.70 % (0.04; 1.37) and hs-CRP increased by 3.16 % (-0.52; 6.98). Adiponectin decreased by -2.53 % (-4.78; -0.24) per 1 IQR (1.4 µg/m3) increase of PM2.5. Besides, PM2.5 was associated with increased IL-6 in the subsample. In conclusion, we observed that long-term exposure to air pollutants, including both fine and ultrafine particles, was associated with higher concentrations of pro-inflammatory and lower concentrations of an anti-inflammatory blood biomarkers, which is consistent with an increased risk for cardiovascular disease observed for long-term exposure to air pollutants.

Microbial consortia and biochar as sustainable biofertilisers: Analysis of their impact on wheat growth and production.

The European Union is among the top wheat producers in the world, but its productivity relies on adequate soil fertilisation. Biofertilisers, either alone or in combination with biochar, can be a preferable alternative to chemical fertilisers. However, the addition of biofertilisers, specifically plant growth promoting microbes (PGPM), could modify grain composition, and/or deteriorate the soil composition. In this study, the two wheat cultivars Triticum aestivum (Bramante) and T. durum (Svevo) were cultivated in open fields for two consecutive years in the presence of a commercial PGPM mix supplied alone or in combination with biochar. An in-depth analysis was conducted by collecting physiological and agronomic data throughout the growth period. The effects of PGPM and biochar were investigated in detail; specifically, soil chemistry and rhizosphere microbial composition were characterized, along with the treatment effects on seed storage proteins. The results demonstrated that the addition of commercial microbial consortia and biochar, alone or in combination, did not modify the rhizospheric microbial community; however, it increased grain yield, especially in the cultivar Svevo (increase of 6.8 %-13.6 %), even though the factors driving the most variations were associated with both climate and cultivar. The total gluten content of the flours was not affected, whereas the main effect of the treatments was a variation in gliadins and low-molecular-weight-glutenin subunits in both cultivars when treated with PGPM and biochar. This suggested improved grain quality, especially regarding the viscoelastic properties of the dough, when the filling period occurred in a dry climate. The results indicate that the application of biofertilisers and biochar may aid the effective management of sustainable wheat cultivation, to support environmental health without altering the biodiversity of the resident microbiome.

Plant responses to climate change, how global warming may impact on food security: a critical review.

Global agricultural production must double by 2050 to meet the demands of an increasing world human population but this challenge is further exacerbated by climate change. Environmental stress, heat, and drought are key drivers in food security and strongly impacts on crop productivity. Moreover, global warming is threatening the survival of many species including those which we rely on for food production, forcing migration of cultivation areas with further impoverishing of the environment and of the genetic variability of crop species with fall out effects on food security. This review considers the relationship of climatic changes and their bearing on sustainability of natural and agricultural ecosystems, as well as the role of omics-technologies, genomics, proteomics, metabolomics, phenomics and ionomics. The use of resource saving technologies such as precision agriculture and new fertilization technologies are discussed with a focus on their use in breeding plants with higher tolerance and adaptability and as mitigation tools for global warming and climate changes. Nevertheless, plants are exposed to multiple stresses. This study lays the basis for the proposition of a novel research paradigm which is referred to a holistic approach and that went beyond the exclusive concept of crop yield, but that included sustainability, socio-economic impacts of production, commercialization, and agroecosystem management.

Ultra-high Performance Liquid Chromatography-Ion Mobility-High-Resolution Mass Spectrometry to Evaluate the Metabolomic Response of Durum Wheat to Sustainable Treatments.

Sustainable agriculture aims at achieving a healthy food production while reducing the use of fertilizers and greenhouse gas emissions using biostimulants and soil amendments. Untargeted metabolomics by ultra-high performance liquid chromatography-ion mobility-high-resolution mass spectrometry, operating in a high-definition MSE mode, was applied to investigate the metabolome of durum wheat in response to sustainable treatments, i.e., the addition of biochar, commercial plant growth promoting microbes, and their combination. Partial least squares-discriminant analysis provided a good discrimination among treatments with sensitivity, specificity, and a non-error rate close to 1. A total of 88 and 45 discriminant compounds having biological, nutritional, and technological implications were tentatively identified in samples grown in 2020 and 2021. The addition of biochar-biostimulants produced the highest up-regulation of lipids and flavonoids, with the glycolipid desaturation being the most impacted pathway, whereas carbohydrates were mostly down-regulated. The findings achieved suggest the safe use of the combined biochar-biostimulant treatment for sustainable wheat cultivation.

Nanoparticles and biochar with adsorbed plant growth-promoting rhizobacteria alleviate Fusarium wilt damage on tomato and watermelon.

The addition of biochars and nanoparticles with adsorbed Azotobacter vinelandii and Bacillus megaterium alleviated damage from Fusarium infection in both tomato (Solanum lycopersicum) and watermelon (Citrullus lanatus) plants. Tomato and watermelon plants were grown in greenhouse for 28 and 30 days (respectively) and were treated with either nanoparticles (chitosan-coated mesoporous silica or nanoclay) or varying biochars (biochar produced by pyrolysis, gasification and pyrogasification). Treatments with nanoparticles and biochars were applied in two variants - with or without adsorbed plant-growth promoting bacteria (PGPR). Chitosan-coated mesoporous silica nanoparticles with adsorbed bacteria increased chlorophyll content in infected tomato and watermelon plants (1.12 times and 1.63 times, respectively) to a greater extent than nanoclay with adsorbed bacteria (1.10 times and 1.38 times, respectively). However, the impact on other endpoints (viability of plant cells, phosphorus and nitrogen content, as well antioxidative status) was species-specific. In all cases, plants treated with adsorbed bacteria responded better than plants without bacteria. For example, the content of antioxidative compounds in diseased watermelon plants increased nearly 46% upon addition of Aries biochar and by approximately 52% upon addition of Aries biochar with adsorbed bacteria. The overall effect on disease suppression was due to combination of the antifungal effects of both nanoparticles (and biochars) and plant-growth promoting bacteria. These findings suggest that nanoparticles or biochars with adsorbed PGPR could be viewed as a novel and sustainable solution for management of Fusarium wilt.

Immunomodulatory peptides-A promising source for novel functional food production and drug discovery.

Immunomodulatory peptides are a complex class of bioactive peptides that encompasses substances with different mechanisms of action. Immunomodulatory peptides could also be used in vaccines as adjuvants which would be extremely desirable, especially in response to pandemics. Thus, immunomodulatory peptides in food of plant origin could be regarded both as valuable suplements of novel functional food preparation and/or as precursors or possible active ingredients for drugs design for treatment variety of conditions arising from impaired function of immune system. Given variety of mechanisms, different tests are required to assess effects of immunomodulatory peptides. Some of those effects show good correlation with in vivo results but others, less so. Certain plant peptides, such as defensins, show both immunomodulatory and antimicrobial effect, which makes them interesting candidates for preparation of functional food and feed, as well as templates for design of synthetic peptides.

Building a risk matrix for the safety assessment of wood derived biochars.

Biochar is recognized as an efficient amendment and soil improver. However, environmental and quality assessments are needed to ensure the sustainability of its use in agriculture. This work considers the biochar's chemical-physical characterization and its potential phyto- and geno-toxicity, assessed with germination and Ames tests, obtaining valuable information for a safe field application. Three biochar types, obtained from gasification at different temperatures of green biomasses from the Tuscan-Emilian Apennines (in Italy), were compared through a broad chemical, physical and biological evaluation. The results obtained showed the relevance of temperature in determining the chemical and morphological properties of biochar, which was shown with several analytical techniques such as the elemental composition, water holding capacity, ash content, but also with FTIR and X-ray spectroscopies. These techniques showed the presence of different relevant surface aliphatic and aromatic groups. The procedures for evaluating the potential toxicity using seeds germination and Ames genotoxicity assay highlights that biochar does not cause detrimental effects when it enters in contact with soil, micro- and macro-organisms, and plants. The genotoxicity test provided a new highlight in evaluating biochar environmental safety.

Effectiveness of a Protocol to Reduce Children's Exposure to Particulate Matter and NO2 in Schools during Alert Days.

Reducing children's exposure to air pollutants should be considered a primary goal, especially for the most vulnerable subjects. The goal of this study was to test the effectiveness of applying a protocol in the event of alert days, i.e., days with forecasted PM10 levels above the EU limit value (50 µg/m3). The test was conducted, before the onset of SARS-CoV-2 restrictions, in a classroom of a primary school in Parma (Italy)-a highly polluted area in Northern Italy. The protocol included indications for the frequency of opening windows and doors, as well as the activation of an air purifier. Teachers and students were asked to apply the protocol only in the event of alert days, while no indications were provided for non-alert days. A monitoring system measuring PM1, PM2.5, PM10, CO2, and NO2 was deployed in the classroom. Measurements of the same parameters were also performed outdoors near the school. The application of the protocol reduced the indoor/outdoor (I/O) ratio for all toxic pollutants. The reduction was also remarkable for PM10-the most critical air quality parameter in the study area (1.5 and 1.1 for non-alert and alert days, respectively). Indoor concentrations of PM10-especially during non-alert days-were often higher than outdoors, showing a major contribution from resuspension due to the movement of people and personal cloud. The protocol did not cause any increase in indoor CO2 levels. Our findings showed that the application of a ventilation protocol together with the contribution of an air purifier may represent an effective way to reduce children's exposure to air pollution during severe air pollution episodes. Considering the onset of COVID-19 and the airborne transmission of pathogens, this protocol now has more meaningful implications for children's welfare, and can be integrated with protocols designed as measures against the spread of SARS-CoV-2.

Engineered Nanoparticles, Natural Nanoclay and Biochar, as Carriers of Plant-Growth Promoting Bacteria.

The potential of biochar and nanoparticles to serve as effective delivery agents for beneficial bacteria to crops was investigated. Application of nanoparticles and biochar as carriers for beneficial bacteria improved not only the amount of nitrogen-fixing and phosphorus-solubilizing bacteria in soil, but also improved chlorophyll content (1.2-1.3 times), cell viability (1.1-1.5 times), and antioxidative properties (1.1-1.4 times) compared to control plants. Treatments also improved content of phosphorus (P) (1.1-1.6 times) and nitrogen (N) (1.1-1.4 times higher) in both tomato and watermelon plants. However, the effect of biochars and nanoparticles were species-specific. For example, chitosan-coated mesoporous silica nanoparticles with adsorbed bacteria increased the phosphorus content in tomato by 1.2 times compared to a 1.1-fold increase when nanoclay with adsorbed bacteria was applied. In watermelon, the situation was reversed: 1.1-fold increase in the case of chitosan-coated mesoporous silica nanoparticles and 1.2 times in case of nanoclay with adsorbed bacteria. Our findings demonstrate that use of nanoparticles and biochar as carriers for beneficial bacteria significantly improved plant growth and health. These findings are useful for design and synthesis of novel and sustainable biofertilizer formulations.

Correlating SNP genotype with the phenotypic response to exposure to cadmium in Populus spp.

Species within the genus Populus include potential phytoextractors of heavy metal ions from contaminated soils, and genetic markers predictive of performance would be a useful tool for selection and breeding. Here, we have identified sequence variation within seven target and three nontarget genes among a set of 11 Populus spp. clones. Sequence variants were present in both the coding and noncoding regions; the former can potentially affect the functionality of the target genes. At the same time, the effect of exposure of the clones to cadmium ions on the morphology and the distribution of various metal ions was investigated by scanning electron microscopy microanalysis. A positive correlation was established between genetic variation, cadmium accumulation, and its bioconcentration in the root.

Identification of Beneficial Microbial Consortia and Bioactive Compounds with Potential as Plant Biostimulants for a Sustainable Agriculture.

A growing body of evidence demonstrates the potential of various microbes to enhance plant productivity in cropping systems although their successful field application may be impaired by several biotic and abiotic constraints. In the present work, we aimed at developing multifunctional synthetic microbial consortia to be used in combination with suitable bioactive compounds for improving crop yield and quality. Plant growth-promoting microorganisms (PGPMs) with different functional attributes were identified by a bottom-up approach. A comprehensive literature survey on PGPMs associated with maize, wheat, potato and tomato, and on commercial formulations, was conducted by examining peer-reviewed scientific publications and results from relevant European projects. Metagenome fragment recruitments on genomes of potential PGPMs represented in databases were also performed to help identify plant growth-promoting (PGP) strains. Following evidence of their ability to coexist, isolated PGPMs were synthetically assembled into three different microbial consortia. Additionally, the effects of bioactive compounds on the growth of individually PGPMs were tested in starvation conditions. The different combination products based on microbial and non-microbial biostimulants (BS) appear worth considering for greenhouse and open field trials to select those potentially adoptable in sustainable agriculture.

Methods for detection of GMOs in food and feed.

This paper reviews aspects relevant to detection and quantification of genetically modified (GM) material within the feed/food chain. The GM crop regulatory framework at the international level is evaluated with reference to traceability and labelling. Current analytical methods for the detection, identification, and quantification of transgenic DNA in food and feed are reviewed. These methods include quantitative real-time PCR, multiplex PCR, and multiplex real-time PCR. Particular attention is paid to methods able to identify multiple GM events in a single reaction and to the development of microdevices and microsensors, though they have not been fully validated for application.

Structural and Functional Features of Chars From Different Biomasses as Potential Plant Amendments.

Biochars result from the pyrolysis of biomass waste of plant and animal origin. The interest in these materials stems from their potential for improving soil quality due to increased microporosity, carbon pool, water retention, and their active capacity for metal adsorption from soil and irrigation water. Applications in agriculture have been studied under different conditions, but the overall results are still unclear. Char structure, which varies widely according to the pyrolysis process and the nature of feedstock, is thought to be a major factor in the interaction of chars with soil and their metal ion adsorption/chelation properties. Furthermore, biochar nutrients and their elemental content can modify soil fertility. Therefore, the use of biochars in agricultural settings should be examined carefully by conducting experimental trials. Three key problems encountered in the use of biochar involve (i) optimizing pyrolysis for biomass conversion into energy and biochar, (ii) physicochemically characterizing biochar, and (iii) identifying the best possible conditions for biochar use in soil improvement. To investigate these issues, two types of wood pellets, plus digestate and poultry litter, were separately converted into biochar using different technologies: pyrolysis/pyrogasification or catalytic (thermo)reforming. The following physicochemical features for the different biochar batches were measured: pH, conductivity, bulk density, humidity and ash content, particle size, total organic substances, and trace element concentrations. Fine porous structure analysis and total elemental analysis were performed using environmental scanning electron microscopy along with energy-dispersive X-ray spectrometry (EDX). Phytotoxicity tests were performed for each biochar. Finally, we were able to (i) differentiate the biochars according to their physicochemical properties, microstructure, elemental contents, and original raw biomass; (ii) correlate the whole biochar features with their respective optimal concentrations when used as plant fertilizers or soil improvers; and (iii) show that biochars from animal origin were phytotoxic at lower concentrations than those from plant feedstock.