This meta-analysis systematically reviews the genetic and breeding strategies for enhancing spring wheat (Triticum aestivum L.) adaptation to Russia's vast Non-Chernozem Zone (NCZ). The NCZ presents significant agricultural challenges due to its heterogeneous agro-climatic conditions, which are categorized into four distinct regional groups. The analysis, conducted following PRISMA guidelines, synthesizes data from 152 selected studies to identify key genetic targets and evaluate modern breeding methodologies. The findings reveal that genetic improvement for the NCZ hinges on developing resilience to a complex of abiotic and biotic stresses. Critical abiotic targets include alleles for photoperiod insensitivity (Ppd-1) and vernalization (Vrn-1) to ensure timely maturation, alongside genes for cold tolerance (CBF regulon), drought resilience (root architecture QTLs, dehydrins), and heat stability. For biotic stresses, the strategy emphasizes pyramiding both major resistance genes and durable adult plant resistance (APR) complexes, such as Lr34/Yr18 and Fhb1, to combat evolving pathogens like rusts, Fusarium head blight, and powdery mildew. The review underscores a paradigm shift from conventional breeding to integrated approaches that leverage genomic selection (GS), marker-assisted selection (MAS), speed breeding, and high-throughput phenotyping. These tools enable the rapid pyramiding of complex traits and enhance selection efficiency for polygenic characteristics. The conclusion asserts that the future of spring wheat breeding in the NCZ lies in the synergistic application of advanced genomic tools and precise phenotyping within a multi-trait selection framework to develop high-yielding, climate-resilient cultivars tailored to the zone's unique and demanding constraints.

In recent years, sustainable industrial development has been expanding globally. Different industries produce by-products that pose significant environmental challenges. Fly ash is one of the by-products from the coal combustion process used by many industries. For the sustainable utilization of by-products, fly ash has gained increasing attention by improving soil quality, and can also be used as a potential solution for the waste management system. As a soil amendment ingredient, the impact of fly ash-enriched soil on plant growth requires a comprehensive evaluation and understanding. In this research, the growth of the radish plant is analyzed by a Multi-Attribute Decision-Making (MADM) method. The growth data of the radish plant were collected under two types of soil conditions with different fly-ash compositions. The experiment was performed under semi-controlled conditions in a greenhouse facility. In this research, we used a multi-objective optimization framework, which is one of the MADM methods, to analyze the impact of the fly ash composition in the delta clay-rich soil and coastal sandy soil on plant growth. The data for five fly ash compositions: 5%, 10%, 15%, and 20% were collected for the delta clay-rich soil and coastal sandy soil. The significant growth factors are germination rate, plant height, and number of leaves. We used the 'Multi-Objective Optimization on the Basis of Ratio Analysis' (MOORA) technique to understand the most significant configuration for evaluating the plant growth under different soil conditions. The MOORA technique is widely used in process parameters optimization, materials selection, and system performance assessment. The method simultaneously incorporates both desirable and undesirable criteria by using a ratio system to convert a decision matrix into a ranking system based on the sample configuration. Our research finds the optimized fly ash composition and the impacts of soil conditions on the radish plant growth in different soil conditions.

The rising demand for sustainable and eco-friendly alternatives to chemical pesticides has intensified research into the potential of insect-derived by-products for plant protection. This study investigates the antifungal efficacy of Tenebrio molitor frass against two major soil-borne pathogens, Rhizoctonia solani and Fusarium oxysporum f.sp. lycopersici, through a two-tiered experimental approach: in vitro and in planta assays using tomato plants (Solanum lycopersicum, cv. ‘Optima F1’).

In vitro experiments evaluated both liquid and solid forms of frass for their ability to inhibit fungal growth. Liquid extracts—both autoclaved and non-autoclaved—were tested via disk diffusion and direct application methods, using multiple controls, including commercial fungicide and pH-adjusted solutions. Significant growth inhibition was observed in R. solani when treated with frass extracts, while F. oxysporum remained largely unaffected. Solid media assays using PDA amended with 1% frass corroborated these findings, showing a 38.06% reduction in R. solani growth, with no statistically significant impact on F. oxysporum.

Following these promising in vitro results, in planta assays were conducted using frass-amended substrates to assess disease suppression and plant growth parameters under R. solani infection. Plants grown in the presence of 1% frass exhibited reduced disease symptoms, increased root biomass, and enhanced shoot length. Statistics analysis revealed that frass significantly influenced shoot elongation in both infected (+2.63 cm) and uninfected plants (+5.29 cm), with a more pronounced effect in the absence of the pathogen.

These results suggest that T. molitor frass holds potential as a biological soil amendment with dual benefits: mitigating disease impact and promoting plant growth. Further research is warranted to elucidate the mechanisms underlying its bioactivity and to optimize its application in integrated pest management (IPM) strategies.

Postharvest fungal diseases remain one of the most significant causes of economic losses in pome fruits, leading to substantial waste during storage, distribution, and commercialization. This challenge is becoming increasingly critical due to the instability of current environmental conditions. The progressive global restrictions on synthetic fungicides demand the development of effective, sustainable, and climate-resilient alternatives. In this context, endophytic microorganisms—naturally inhabiting plant tissues without causing harm—represent a promising, underexplored reservoir of biological diversity with high potential for the discovery of novel biocontrol agents and antifungal metabolites applicable to postharvest protection.

In this study, a broad collection of 88 endophytic isolates (65 fungi/yeasts and 23 bacteria) was obtained from apple, pear and nashi fruits belonging to several commercial varieties. Microorganisms were isolated from both mesocarp and peduncle tissues, with the aim of capturing a representative diversity of fruit-associated endophytes. The antagonistic potential of all isolates was systematically screened through a series of in vitro assays against five major postharvest fungal pathogens of pome fruit: Penicillium expansum, Botrytis cinerea, Alternaria alternata, Colletotrichum acutatum and Monilinia fructigena. Initial confrontation assays revealed that multiple isolates were capable of reproducibly inhibiting pathogen development, as quantified through reductions in colony area relative to untreated controls. These results permitted the selection of a subset of promising strains for further mechanistic studies.

To gain insight into potential modes of action, split-plate assays were performed to evaluate volatile-mediated inhibition. No significant growth suppression was observed, suggesting that volatile organic compounds (VOCs) were not responsible for the antagonistic activity detected in earlier assays. Conversely, liquid-culture filtrates obtained from selected strains revealed that at least one endophyte produced extracellular antifungal metabolites capable of reducing pathogen growth in vitro, supporting the hypothesis that bioactive, secreted compounds contribute to its inhibitory activity. This finding identifies the strain as a particularly strong candidate for future metabolite characterization.

Current and future work is focused on the molecular identification of the most active isolates, extraction and characterization of their inhibitory metabolites, and the assessment of additional mechanisms potentially involved in antagonism, including competition and nutrient depletion. Furthermore, the research pipeline will progress toward in vivo validation on fresh fruit to determine the real-world applicability of the most promising candidates.

Overall, this study highlights the potential of fruit-associated endophytes as a valuable source of sustainable biocontrol agents and antifungal compounds, contributing to the development of climate-adaptive postharvest management strategies and offering new tools for reducing fruit losses in a more environmentally conscious framework.

Salinity stress is a major constraint to global crop productivity, affecting physiological processes and reducing yield and quality in many horticultural crops. Although tomato (Solanum lycopersicum L.) is moderately salt-sensitive, salt-induced reductions in photosynthetic capacity, fruit size, and overall plant performance remain significant challenges for sustainable production. Endophytic fungi produce a diverse array of bioactive fungal extracts with potential biostimulant properties. This study evaluated whether foliar application of Talaromyces ruber extracts could alleviate salinity stress effects on tomato plants and improve fruit quality parameters. Tomato plants were grown in a climate-controlled greenhouse with four treatments established: control, salinity, fungal extracts and salinity + fungal extracts. Foliar application of fungal extracts were carried out at two developmental stages: early fruit set and full flowering. Physiological parameters, chlorophyll content, flavonol index, and anthocyanin index, were measured at three time points. Furthermore, yield components, fruit diameter, soluble solids content (°Brix), and shoot biomass were also evaluated. The interaction between salinity and fungal extract application significantly affected chlorophyll content during the first evaluation period, indicating that extracts enhanced chlorophyll accumulation specifically under saline conditions. Although salinity drastically reduced total fruit production per plant, the application of fungal extracts significantly improved fruit diameter regardless of salinity level, with mean fruit diameter increasing in extract-treated plants under both control and saline conditions. This suggests a positive effect on commercial fruit quality, although extracts did not significantly enhance total yield or shoot biomass. While the extracts tested did not confer tolerance to high salinity levels, their ability to improve fruit calibre highlights their potential as biostimulants in less extreme saline environments or when applied more frequently throughout the crop cycle. Future research should explore lower salinity levels, repeated applications, and identification of specific bioactive compounds responsible for the observed effects, contributing to the growing evidence supporting endophytic fungi as sources of sustainable, eco-friendly biostimulants for stress management in horticulture.

Chitinases are vital to several physiological processes in insects such as growth and development, digestion, and even immune response. The coffee berry borer (CBB, Hypothenemus hampei) is regarded as a major pest directly infesting the economically important berry of the coffee plant. Yet unexplored, functions of chitinases in CBB offer avenues for further study. Here, 19 chitinase and chitinase-like genes were identified in CBB (HhCHTs) through BLAST-based homology search and conserved motif analysis. Phylogenetic analysis grouped these HhCHTs into seven clades, together with chitinases from other insect taxa. The HhCHTs exhibited coding sequences of 864-7998 bp which corresponded to protein lengths of 287-2665 amino acids. The HhCHTs showed an average isoelectric point of 5.97 and an average molecular weight of 82.2 kDa. Physicochemical analysis indicated that most proteins are stable, hydrophilic, and thermostable. Domain and motif analyses revealed the conserved structures among the genes, including the presence of GH18 catalytic domain (PF00704) and CBM14 chitin-binding domain. Subcellular localization prediction using CELLO v.2.5 suggested that HhCHT-03, -06, -07, -11, and -13 are extracellular which imply their roles in cuticle formation or remodeling. Protein interaction networks revealed a highly interconnected network, with several HhCHTs (e.g., HhCHT-08, -09, -10, -11, -15, -17, and -19) interacting closely with β-hexosaminidases and proteins containing GH20 or GH18 domains. These associations point to shared functions in glucosamine-containing compound catabolism, amino sugar and nucleotide sugar metabolism, glycosphingolipid biosynthesis, and chitinase-related activity. Additionally, protein structure modeling and predictions of transmembrane helices and signal peptides further clarified the structural and functional features of the identified chitinases. Overall, this study provides the first comprehensive characterization of the chitinase gene family in the coffee berry borer, offering foundational insights for the development of molecular pest management strategies, including RNA interference-based control.

Synthetic herbicides remain the predominant management strategy for agricultural weeds, yet their widespread application has engendered environmental concerns and the emergence of resistant populations. The investigation of plant-derived allelopathic compounds offers a viable alternative for developing biodegradable and ecologically sustainable weed control agents. Ridolfia segetum (L.) Moris, arable weed belonging to the Apiaceae family, harbors diverse secondary metabolites with established phytotoxic properties. The phytotoxic efficacy of aqueous extracts derived from R. segetum tissues against Sinapis alba (white mustard), a prevalent cruciferous weed species problematic in cereal and legume cropping systems, was evaluated through systematic laboratory and greenhouse experiments. Fresh aerial parts of R. segetum were collected at the flowering stage and subjected to aqueous extraction at plant-to-water ratios of 1:10, 1:20, and 1:40 (w/v). The aqueous extraction methodology, encompassing various solvent concentrations and extraction temperatures (25°C, 50°C, and 80°C for 24, 48, and 72 hours), significantly influences the release and bioavailability of allelopathic compounds, including coumarins, volatile oils, and phenolic derivatives. Phytotoxic evaluations conducted across multiple growth stages reveal dose-dependent inhibitory effects on seed germination rates, radicle elongation, and seedling biomass accumulation in S. alba. At concentrations of 25%, 50%, 75%, and 100% (v/v), the extracts reduced seed germination by 23%, 47%, 68%, and 89% respectively compared to controls. Radicle length was suppressed by 31-92%, while shoot length decreased by 28-85% across the concentration gradient. Fresh biomass accumulation showed reductions of 35-94% relative to untreated seedlings. Mechanistic investigations indicate that R. segetum aqueous extracts suppress photosynthetic efficiency and disrupt oxidative stress homeostasis in target seedlings through elevated reactive oxygen species production and diminished antioxidant enzyme activities. Chlorophyll content decreased by 42-78%, while measurements of malondialdehyde levels increased by 2.3 to 4.8-fold, indicating lipid peroxidation. Activities of catalase, peroxidase, and superoxide dismutase were reduced by 34-71%, 29-66%, and 38-73% respectively at higher extract concentrations. Temporal assessments demonstrate that extract potency remains relatively stable over short-term storage periods (up to 15 days at 4°C with 8-12% activity loss), though degradation of bioactive components occurs under extended conditions, with approximately 45-60% reduction in phytotoxic activity after 60 days. Comparative analysis with existing bioherbicide formulations demonstrates competitive or superior suppression of S. alba growth at equivalent application rates, with R. segetum extracts achieving 82-91% weed suppression compared to 74-86% for commercial alternatives. Field-based assessments indicate variable efficacy under natural conditions, contingent upon soil edaphic properties, prevailing moisture regimes, and the developmental stage of target weeds. Plot trials across three soil types (sandy loam, clay loam, and silty clay) showed weed biomass reductions of 56%, 68%, and 71% respectively at 14 days after treatment. This work synthesize available evidence regarding optimal extraction protocols, identifies knowledge gaps concerning the identity and synergistic interactions of active phytotoxic compounds, and discusses the regulatory and practical considerations inherent to bioherbicide commercialization. Integration of R. segetum-derived allelopathic preparations into conservation agriculture systems and organic production frameworks could substantially reduce herbicide dependency whilst maintaining agronomic productivity. Future research priorities encompassing field-scale validation across diverse edaphoclimatic contexts, standardization of extraction methodologies, and characterization of phytotoxic mechanisms at the molecular level are essential for advancing this biotechnological approach toward practical agriculture implementation.

Endophytic fungi represent a largely untapped reservoir of bioactive secondary metabolites with potential applications in sustainable agriculture. The discovery of novel biopesticides from these symbionts has been hindered by traditional screening methods that fail to capture the full biosynthetic capacity of fungal genomes. This study employed an integrated multi-omics strategy combining genomics, transcriptomics, metabolomics, and proteomics to systematically identify and characterize pesticidal compounds from endophytic fungi isolated from medicinal plants. Fungal endophytes were recovered from surface-sterilized plant tissues and authenticated through morphological examination and molecular phylogeny using multi-locus sequence typing. A total of 47 endophytic isolates were obtained from Azadirachta indica, Withania somnifera, and Ocimum sanctum, with 23 isolates selected for detailed analysis based on preliminary bioactivity screening. Whole-genome sequencing was performed using PacBio long-read technology to identify biosynthetic gene clusters associated with polyketide synthases, non-ribosomal peptide synthetases, and terpene cyclases. Genome analysis revealed 58 to 72 biosynthetic gene clusters per isolate, with an average genome size of 42.3 Mb. Transcriptomic profiling under various culture conditions revealed differential expression patterns of biosynthetic genes, with approximately 38% of identified gene clusters showing significant upregulation under stress conditions, while LC-MS/MS-based metabolomics facilitated the annotation of 523 secondary metabolites. Proteomic analysis confirmed the translation of key enzymatic machinery involved in secondary metabolism, identifying 847 proteins with 156 directly linked to specialized metabolite production. Bioactivity-guided fractionation of fungal extracts was conducted against agriculturally important pests, including Helicoverpa armigera, Spodoptera litura, and Tetranychus urticae, as well as phytopathogenic fungi such as Fusarium oxysporum and Botrytis cinerea. Testing revealed that 12 isolated fractions exhibited insecticidal activity with LC₅₀ values ranging from 18 to 165 μg/mL, while eight fractions showed antifungal properties with minimum inhibitory concentrations between 4.2 and 32 μg/mL. Integration of omics datasets through bioinformatics pipelines enabled the correlation of specific biosynthetic pathways with observed pesticidal activities. Several previously unreported compounds were identified, including modified polyketides and cyclic peptides exhibiting nanomolar-range insecticidal and fungicidal properties. Three lead compounds demonstrated LC₅₀ values of 23 nM, 67 nM, and 142 nM against H. armigera larvae. Mode of action studies using electrophysiology and enzymatic assays revealed interference with neuronal acetylcholinesterase activity (IC₅₀ = 15-34 nM) and disruption of fungal cell wall biosynthesis through β-1,3-glucan synthase inhibition (IC₅₀ = 2.8-8.6 μM). Genome mining identified silent gene clusters that were activated through epigenetic modulation using histone deacetylase inhibitors, expanding the chemical diversity accessible from these organisms by yielding 18 additional compounds not detected under standard cultivation. Quantitative structure-activity relationship modeling was applied to optimize lead compounds for enhanced stability and reduced mammalian toxicity, resulting in derivatives with 3.5 to 6-fold improved photostability and 40-60% reduced toxicity in mammalian cell lines. The compounds demonstrated favorable environmental profiles with rapid photodegradation (t₁/₂ = 6-14 hours under natural sunlight) and minimal non-target effects in preliminary ecotoxicological assessments, showing negligible toxicity to Daphnia magna (LC₅₀ > 500 mg/L) and honeybees (LD₅₀ > 100 μg/bee). This comprehensive multi-omics framework accelerates the pipeline from endophyte isolation to biopesticide. The findings highlight the potential of endophytic fungi as sources of next-generation biopesticides and provide molecular insights into fungal specialized metabolism relevant to agrochemical development.

Abstract: The sweet chestnut (Castanea sativa Mill.) is a crop of significant socioeconomic and ecological importance in Portugal. However, its production is heavily affected by several pests, notably chestnut weevils and moths (locally known as bichados), including Cydia fagiglandana, C. splendana, Curculio elephas, and Pammene fasciana. The continuous use of chemical insecticides to control these pests raises environmental and public health concerns, highlighting the urgent need to develop sustainable and eco-friendly alternatives. Entomopathogenic fungi (EPF) represent a promising biological tool capable of selectively reducing pest populations, in line with European Union policies aimed at decreasing agrochemical inputs. Herein, this study aimed to isolate, identify, and assess the diversity of EPF from chestnut grove soils in experimental plots located in Trancoso, as well as to evaluate their field efficacy against chestnut pests compared with the commercial insecticides Affirm Opti® and Spintor 480 SC®. EPF were isolated using the insect-bait method with Tenebrio molitor larvae and identified based on morphological traits and molecular analysis of the ITS and TEF1 regions, followed by phylogenetic analyses. The results revealed a high diversity and abundance of EPF in the chestnut grove soils, with six species isolated. Metarhizium robertsii was the most frequently isolated species, followed by Beauveria bassiana. In field trials, strains of these two predominant species significantly reduced chestnut fruit infestation compared with the chemical treatment. Notably, B. bassiana achieved 100% efficacy against all evaluated pests, followed by M. robertsii, whereas the chemical treatment (Affirm Opti®) exhibited the lowest efficacy. Overall, the native EPF isolates demonstrated strong potential in controlling chestnut pests, particularly B. bassiana, supporting their integration into sustainable and eco-friendly pest management strategies. Further studies are underway to optimize formulation and application methods to facilitate the development of effective commercial bioinsecticides.

Acknowledgments: This work was supported by the project 101074466 — LIFE21-CCA-IT-LIFE FAGESOS and by National Funds through the Fundação para a Ciência e a Tecnologia (FCT) under the scope of the projects UID/04033/2025 (CITAB) (https://doi.org/10.54499/UID/04033/2025) and LA/P/0126/2020 (Inov4Agro) (https://doi.org/10.54499/LA/P/0126/2020). EHE thanks the PRR and the European Funds NextGeneration EU for the research grant BIPD/UTAD/3/2023.

Fruit crops like guava, pomegranate are good hosts to root knot nematodes, but only few studies were previously conducted for management of root knot nematodes. Pomegranate is a perennial crop which is a host to root knot nematodes, can harbour high nematode populations causing heavy yield losses and death of plants by secondary infection of soil borne pathogens. This is the first study where the bio control potential of P. lilacinum, P. chlamydosporia, and B. subtilis at different doses were explored to manage Meloidogyne incognita infestations in pomegranate at two locations, Haryana and Rajasthan for three consecutive years. Results revealed that both the fungal and bacterial biocontrol agents effectively reduced nematode galling and the overall nematode population in soil while concurrently boosting pomegranate fruit yield. The number of galls remained consistently high in control plants and was second highest in plants treated with neem cake. Treatment with B. subtilis exhibited the lowest gall count (225-99), while remaining treatments also showed fewer galls, particularly in later years. Moreover, in Rajasthan, the number of galls per 5 grams of roots was lower than in Rohtak, with B. subtilis showing the lowest gall index, especially in the third year. Over time, all the treatments showed progressive increase in plant height across time i.e. June 2021 to June 2024. B. subtilis at highest dose achieved maximum height at every point from December 2021 onwards. Also in both the locations plant width expanded greatly in plants, treated with B. subtilis exhibiting the significant difference from the control plants.