Search Results (494)

This study investigated the effects of repeated dielectric barrier discharge (DBD) plasma applications on the morphological and physiological characteristics of pathogenic Alternaria and Fusarium species. Fungi, including both culture collection strains and environmental isolates, were exposed to sublethal doses of DBD plasma. The results demonstrated that the plasma exposure time required to achieve 90% cell mortality varied significantly among microorganisms, ranging from 2 min and 39 s for Fusarium culmorum DSM 1094 to 5 min and 19 s for Alternaria alternata DSM 62010. Tolerance to oxidative stress, assessed by determining the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of hydrogen peroxide, generally decreased following repeated plasma exposure. Notably, F. tricinctum Ft11S–23 exhibited increased resistance to hydrogen peroxide, with MIC values doubling after fifteen plasma treatments. The MFC also increased significantly, rising from 25.5 mM to 102.0 mM. Furthermore, repeated DBD plasma applications resulted in reduced tolerance of fungi to at least one of the tested fungicides; however, exceptions were observed, including increased tolerance of F. culmorum to specific fungicides. The capacity for biofilm formation was modulated by plasma treatment, with some species exhibiting reduced biofilm formation while others demonstrated increased capacity, depending on the specific pathogen and frequency of plasma exposure. Full article

Apple (Malus domestica Borkh.) production is increasingly threatened by fungal diseases under conditions of intensive horticulture and ongoing climate change. In southeastern Kazakhstan, symptoms associated with Alternaria spp. have become more frequent in commercial orchards; however, molecularly confirmed data on the pathogen associated with these symptoms remain limited. This study aimed to identify, using multigene molecular phylogenetic analysis, an Alternaria isolate obtained from infected apple leaves and fruits, to confirm its pathogenicity experimentally, and to compare the susceptibility of apple cultivars to this pathogen. For molecular identification, nucleotide sequences of four genetic markers—the ITS region of rDNA, SSU, tef1-α, and RPB2—were obtained by PCR and sequencing. The sequences were compared with reference data from the GenBank database using BLASTn, and phylogenetic relationships were inferred using the maximum likelihood method based on a concatenated dataset of the four loci. The isolate KZ17 clustered within the A. alternata clade, with the broader node uniting this group with A. gossypina and A. longipes receiving a bootstrap value of 78%. A total of 20 fungal isolates were obtained from 112 symptomatic apple leaf and fruit samples. Among them, one representative isolate, KZ17, was selected for multilocus molecular identification and pathogenicity testing. The pathogenicity of isolate KZ17 was confirmed in inoculation experiments on apple microshoots under controlled conditions. Artificial inoculation of detached leaves and ripe fruits of 14 apple cultivars revealed significant cultivar-dependent differences in susceptibility. Lesion diameters ranged from 12.3 ± 0.20 to 19.2 ± 0.35 mm on detached leaves and from 15.0 ± 1.2 to 30.0 ± 2.4 mm on ripe fruits. The least susceptible cultivars were ‘Granny Smith’, ‘Zaman’, and ‘Maksat’, whereas ‘Voskhod’, ‘Saltanat’, and ‘Kamila’ showed the greatest susceptibility. These differences were statistically significant (p < 0.001). This study provides the first multigene-based confirmation of the association of A. alternata with apple leaf and fruit lesions in southeastern Kazakhstan and demonstrates cultivar-dependent differences in susceptibility to this pathogen. These findings contribute to improved pathogen diagnostics, germplasm screening for resistance, and the development of plant protection strategies for commercial apple orchards. Full article

Sonoplasma treatment (SPT), which combines hydrodynamic cavitation with low-temperature plasma discharge in water, has been proposed as an advanced oxidation process for reducing biological contamination. By generating physical stressors and reactive oxygen species, including hydrogen peroxide (HP), SPT may inactivate microorganisms, but its effects on stress-resistant filamentous fungi remain insufficiently characterized. We examined two phytopathogenic fungi with contrasting pigmentation: melanized Alternaria alternata and hyaline Fusarium solani. Spore suspensions were exposed to direct and indirect SPT at 30 kHz, and viability, biomass accumulation, conidial production, allelopathic activity, and pigmentation-associated spectral responses were assessed immediately after treatment and after storage. Fungus F. solani showed greater susceptibility, with reduced colony-forming capacity and suppressed biomass production, although surviving propagules showed increased sporulation. In contrast, A. alternata maintained viable growth under the tested conditions and showed stimulation of growth-related and reproductive endpoints, together with darker colony pigmentation. These responses are consistent with pigmentation-associated tolerance to SPT-induced physical and oxidative stress, but do not establish melanin as the sole causal mechanism. SPT efficacy against filamentous fungi is therefore species-dependent and may be limited when resistant melanized taxa are present. Full article

Ginseng Alternaria leaf and stem blight (GALSB), caused by Alternaria alternata, poses a severe threat to ginseng cultivation. Although azoxystrobin is a cornerstone fungicide for GALSB management, the emergence of widespread adaptive resistance has severely curtailed its field efficacy. This study integrated molecular, transcriptomic, and genetic approaches to unravel the underlying resistance mechanisms. Targeted gene sequencing and molecular docking revealed that resistant strains harbor a conserved G143A point mutation in the AaCyt b protein. This mutation weakens the azoxystrobin–AaCyt b protein binding affinity by decreasing the binding energy from −8.31 to −7.08 kcal/mol. Additionally, comparative transcriptomics and RT-qPCR demonstrated pronounced upregulation of the alternative oxidase gene (AaAOX) and core energy metabolism pathways in resistant strain TYC8-2, with AaAOX expression increasing 4.45–6.91-fold. Fungicidal inhibition of AOX via salicylhydroxamic acid (SHAM) restored fungal sensitivity, increasing azoxystrobin sensitivity by 11.66-fold. Crucially, genetic knockout of AaAOX enhanced sensitivity by approximately 2.7 × 10⁴-fold. Phenotypic assays further established AaAOX as a multifunctional regulator; the AaAOX mutant exhibited attenuated virulence on ginseng leaves and increased sensitivity to oxidative and osmotic stresses (NaCl, H₂O₂, NaAc). We conclude that the G143A mutation in AaCyt b and the transcriptional overexpression of AaAOX act independently to drive azoxystrobin resistance in A. alternata. These findings provide comprehensive mechanistic insights to guide resistance monitoring, optimize fungicide applications, and develop precision strategies for GALSB management. Full article

The increasing demand for sustainable crop protection strategies has intensified interest in plant-beneficial bacteria as alternatives to synthetic agrochemicals. In this study, the soil-derived bacterium Pseudomonas putida V01 was isolated and characterized for its antifungal and plant growth-promoting potential through an integrated approach combining biological assays, untargeted metabolomics, and in vivo plant experiments. Cell-free culture filtrates exhibited strong antifungal activity against major phytopathogenic fungi, completely inhibiting the growth of Sclerotium rolfsii and significantly reducing mycelial development of Alternaria alternata and Fusarium proliferatum by 40% and 20%, respectively. Volatile organic compounds (VOCs) selectively inhibited Botrytis cinerea and A. alternata by 28% and 10%, respectively, and affected sporulation of F. proliferatum. Metabolomic profiling through LC-qTOF-MS and GC-MS analyses revealed a chemically diverse metabolome, including putatively annotated diketopiperazines, cyclic peptides, phenolic compounds, and fatty acids. VOC profiling indicated ketones and alcohols as the predominant volatile classes, with 2-undecanone and 2-undecanol among the most abundant compounds detected. In vivo assays on wheat seedlings showed significant increases in shoot growth, biomass accumulation, and chlorophyll content compared with untreated controls. These findings indicate that P. putida V01 combines complementary antifungal and plant growth-promoting activities associated with a diverse repertoire of diffusible and volatile metabolites. The integrated biological and metabolomic characterization highlights its potential as a multifunctional microbial inoculant for sustainable crop production and disease management. Full article

Thirteen mandarin and hybrid cultivars grafted onto the commercial rootstocks Rangpur Lime and Swingle Citrumelo were comparatively assessed for vegetative growth, fruit physicochemical attributes, and field incidence and severity of Altenaria Brown Spot (ABS) and Huanglongbing (HLB). The experiment was conducted from January 2015 to December 2018 under a randomized block design, with ten replicates per scion–rootstock combination. Plant height, canopy volume, fruit mass, juice yield, acidity, soluble solids, and disease assessments were performed. RL induced greater vegetative growth but was associated with higher HLB severity, particularly in the Dekopon IAC 2009 and TM × LP 358 varieties. SC resulted in less vigorous trees but improved fruit quality, with higher acidity and soluble solids. Regarding ABS, the Loose Jacket IAC 515 and Muscia varieties showed high susceptibility, while Ortanique IAC 554 and Rainha BRS exhibited tolerance to both ABS and HLB. These findings suggest that although RL promotes vigorous growth, it may increase disease susceptibility, whereas SC is associated with reduced disease severity and improved fruit quality. Ortanique IAC 554 and Rainha BRS showed consistently low severity of ABS and HLB, combined with stable vegetative development and fruit quality, underscoring the importance of rootstock choice for guiding cultivar deployment in orchards under high disease pressure. Full article

The present study was conducted to study the effect of exposure time to ultraviolet-C (UV-C) radiation on seed germination, fungal suppression and seedling growth of three Egyptian rice cultivars, namely, Sakha 105, Sakha 108, and Giza 183. Experiments were carried out under controlled laboratory conditions. Rice seeds were exposed to UV-C radiation with a wavelength of 253.7 nm and intensity of 1960 µW cm² for 0 (control), 10, 20, 30, 40, 50, and 60 min. Initial seed health testing showed the presence of several seed-borne fungi, mainly Alternaria alternata, Rhizoctonia solani, and Fusarium verticillioides, in addition to Aspergillus niger and Aspergillus flavus. Results revealed that UV-C exposure time, rice cultivar and their interactions significantly (p < 0.05) affected germination percentage, reduction percentage of seed fungal infection, and seedling growth parameters. The optimum exposure time was 30 min, which was found to maximize germination and improve shoot and root growth to achieve high levels of fungal suppression. Giza 183 exhibited the highest average germination percentage (92.40%), while Sakha 105 obtained the highest shoot height (17.00 cm) and root length (12.91 cm). The results indicate that UV-C irradiation is an effective, residue-free and environmentally sustainable seed treatment technology for improving rice seed quality as well as early seedling performance. Full article

Tomato is susceptible to various fungal pathogens, including Alternaria alternata and Curvularia spicifera, which can cause extensive post-harvest losses. Chemical fungicides have limited effectiveness in controlling post-harvest fungal pathogens and pose risk to human health and the environment. Therefore, this study assessed indigenous isolates of three species of Trichoderma (Tr1: T. longibrachiatum; Tr2: T. harzianum; and Tr3: T. asperellum) as biocontrol agents against two fungal pathogens in vitro and in vivo and determined their physicochemical analysis and plant-growth-promoting traits. The three species of Trichoderma exhibited catalase production in vitro, while T. longibrachiatum and T. asperellum showed the highest potential for plant-growth promotion by producing indole-3-acetic acid and phosphate solubilization but not nitrogen-fixing capability. T. harzianum showed lower potential in these traits. Mycelial growth was found to be maximum (5.77–12.27 cm) at 30 °C and a pH of 7–9, but inhibition (2.60–5.13 cm) was recorded at the highest temperature (45 °C) and pH (11). In vivo, studies on tomato fruits indicated that T. longibrachiatum and T. asperellum significantly (p < 0.05) reduced lesion diameters of A. alternata by 53.60% and 48.71%, respectively, and C. spicifera by 55.58% and 56.19%, respectively, relative to the infected control. Besides their antifungal efficacy, the three species of Trichoderma enhanced tomato seedling growth, particularly at 1/10 filtrate dilution, and improved fruit quality parameters by increasing firmness and nitrate content, while reducing oxidative stress. Physicochemical analysis indicated that Trichoderma-treated fruits had better firmness, pH, and nitrate value coupled with a reduction in oxidative stress (reduced malondialdehyde content) compared to pathogen-infected controls. The indigenous isolates of the three species of Trichoderma provided high efficacy as biocontrol agents of the two fungal pathogens that cause post-harvest losses of tomato, suggesting that biological control can replace synthetic chemicals in preserving tomato under storage conditions and contribute to agricultural sustainability. Full article

To investigate the effect of sorbitol feeding on immune responses and its underlying mechanism, this study employed crabapple (Malus micromalus), a species native to China, as the experimental material. Detached leaves were subjected to sorbitol feeding experiments, with water feeding as the control. The dynamic changes in lesion area and cell membrane permeability were measured across four groups of leaves following feeding and fungal inoculation, and transcriptome sequencing was performed on these four groups. The results revealed that compared to the control group, leaves fed with sorbitol prior to inoculation exhibited significantly reduced lesion areas and decreased cell membrane permeability. Transcriptome and KEGG pathway analyses indicated that differentially expressed genes were significantly enriched in the phenylpropanoid biosynthesis pathway. HPLC results demonstrated that quercetin and isorhamnetin contents increased significantly after sorbitol feeding, and both compounds were shown to inhibit the mycelial growth of Alternaria alternata, thereby enhancing the disease resistance of crabapple leaves. Among the associated genes, MmF3H1, which regulates the synthesis of quercetin and isorhamnetin, showed the highest level of enrichment. Co-expression trend analysis and qPCR results revealed that MmNAC17 and MmbZIP1 exhibited strong co-expression relationships with MmF3H1 and responded to sorbitol regulation. Functional validation was conducted by constructing overexpression and silencing vectors for MmNAC17/MmbZIP1 and performing transient transformation, thereby elucidating the molecular and biochemical mechanisms underlying the response of M. micromalus to A. alternata. This study provides a theoretical reference for breeding crabapple varieties with enhanced resistance to leaf spot disease. Full article

The β-1,3-glucanase gene family plays an important role in fungal cell wall degradation and is closely associated with the mycoparasitic interactions of biocontrol fungi. In this study, the β-1,3-glucanase gene family was identified in the mycoparasitic Alternaria alternata strain KMR13, and its expression patterns under Podosphaera pannosa spore induction were analyzed to investigate the role of the gene Aag25 in the degradation of P. pannosa spores. The β-1,3-glucanase gene was extracted from the genome using the bioinformatics method. According to the expression level of spore-induced genes of P. pannosa, the GH17 family genes that may be involved in mycoparasitism were screened, cloned, and expressed. The expressed protein was purified, and its activity and ability to destroy spores of P. pannosa were determined. A total of 30 β-1,3-glucanase genes were identified in strain KMR13, including 18 members of the GH16 family, 8 of the GH17 family, and 2 genes each from the GH64 and GH81 families. Transcriptome analysis revealed that 23 of these genes were expressed under spore induction. Based on differential expression analysis, the β-1,3-glucanase gene Aag25 was selected for prokaryotic expression. The recombinant Aag25 protein (~32.08 kDa) was successfully induced and purified, exhibiting an enzymatic activity of 1.464 U/mg protein. Functional assays demonstrated that recombinant Aag25 could effectively disrupt the cell walls of P. pannosa spores, leading to spore rupture and cytoplasmic leakage. These results indicate that Aag25 plays an important role in fungal cell wall degradation during the mycoparasitic process of strain KMR13 and provides a potential enzymatic resource for the development of biological control strategies targeting fungal pathogens. Full article

Alternaria alternata causes major postharvest losses in melons. This dual RNA-seq study analyzed transcriptomic dynamics in both A. alternata strain A2022 and melon during infection. Genomic analysis revealed two conditionally dispensable chromosomes (CDCs) containing host-specific toxin genes, including a T1PKS cluster for βenone biosynthesis. Fungal transcriptomics identified upregulated virulence pathways: MAPK signaling (18 genes), autophagy (17 genes), and lipid metabolism. Melon mounted a biphasic defense: early lignin deposition via phenylpropanoid biosynthesis (15 genes) and late jasmonic acid/MAPK-mediated responses. Competitive nutrient acquisition emerged, with fungal amino acid transporters opposing host nitrogen metabolism. These findings reveal A. alternata’s genomic plasticity and melon’s layered resistance, suggesting RNAi-based silencing of CDC virulence genes or breeding for enhanced lignin/jasmonate pathways as sustainable control strategies. Full article

This study aimed to separate and characterize compounds from Aspergillus tubingensis ZMGR14. The antifungal activities of monomer compounds and the ethyl acetate (EtOAc) layer from the fermented liquor of A. tubingensis were isolated, purified and structurally identified. The EtOAc layer from the fermented liquor showed significant antifungal activity against Fusarium oxysporum and Alternaria alternata with IC₅₀ values of 273.8 and 330.7 μg·mL⁻¹, respectively. The EtOAc extract was further purified by column chromatography and recrystallization to yield six compounds. Antifungal trials showed that Cyclo-(L-Pro-D-Leu) (5) exhibited the highest inhibition against A. alternata and F. oxysporum, with an IC₅₀ value of 48.1 and 232.7 μM, respectively, and cyclo-(L-Pro-L-Leu) (6) displayed moderate antifungal activity against Alternaria solani, with an IC₅₀ value of 493.4 μM. The results suggest that the EtOAc extract of ZMGR14 and its bioactive compounds hold promise as environmentally friendly microbial fungicides. Full article

Broccoli (Brassica oleracea var. italica) by-products represent an abundant and underutilized source of bioactive compounds with potential applications in sustainable food systems. This study aimed to characterize the microbiota associated with different plant fractions (leaves, stems, and heads) of broccoli (Parthenon and Tritón cultivars) and to evaluate the antioxidant and antimicrobial properties of their extracts, using cauliflower as a reference. Microbial counts and fungal identification (ITS sequencing) were performed, while phytochemical profiles were analyzed by HPLC-ESI-QTOF. Antioxidant activity was assessed using DPPH and ABTS assays, and antimicrobial activity under in vitro conditions was evaluated against selected foodborne bacteria and phytopathogenic fungi. Broccoli by-products, particularly leaves, showed lower microbial loads in certain cultivars and were rich in phenolic compounds and glucosinolates; however, higher phenolic content did not always correlate with greater antioxidant activity, highlighting the importance of compound composition. All extracts showed strong antibacterial activity at higher concentrations, especially against Listeria spp. Notably, antifungal activity was selective but relevant, with consistent inhibition observed against Alternaria alternata, while Penicillium purpurogenum and Botrytis cinerea exhibited higher resistance. Overall, these findings highlight the potential of broccoli by-products as sustainable sources of natural bioactive compounds for food applications, particularly in the development of preservation strategies and postharvest treatments. Further studies focusing on individual compounds and their specific biological activities are needed to better understand the mechanisms underlying these effects and to support their application in real food systems. Full article

Alternaria blight, caused by fungi of the genus Alternaria, is one of the most common and damaging diseases affecting tomatoes, leading to significant yield losses. The intensive use of chemical fungicides faces the problems of pathogen resistance development and negative environmental impacts. This study investigated the possibility of using RNA interference technology based on exogenous double-stranded RNAs (dsRNAs) to protect tomatoes against the causal agent of early blight (EB), Alternaria alternata. Key genes of the pathogen A. alternata were selected as targets: Alt-a1 (a major allergen and virulence factor), TEF1a (translation elongation factor 1-alpha) and β-Tub (β-tubulin). Specific dsRNAs were synthesized in vitro and applied to tomato plants (Solanum lycopersicum L. cv. Micro-Tom) simultaneously with inoculation of A. alternata strain C7.24-T2-L-F1 spores. Visual assessment, measurement of chlorophyll A and B, and real-time quantitative PCR analysis showed that treatment with dsRNAs targeting the Alt-a1, TEF1a and β-Tub genes significantly suppressed infection development, reducing the amount of pathogen DNA in plant tissues by 7 to 27 times depending on the dsRNA type. The most effective was dsRNA to the Alt-a1 gene. Thus, the obtained results demonstrate the promise of spray-induced gene silencing (SIGS) as a strategy for protecting tomato plants against the pathogen A. alternata. Full article

Endophytic fungi are promising sources of novel antiviral compounds, and the crude extract from Alternaria alternata PO4PR2 has previously shown anti-HIV-1 activity. This study evaluated its efficacy against integrase strand-transfer inhibitor (INSTI)-resistant HIV-1 and its mechanism of action. Key resistance mutations (Y143H, G118R, N155H, and R263K) were introduced into the HIV-1 pNL4.3 clone via site-directed mutagenesis and confirmed through Sanger sequencing. Viral infectivity was assessed in TZM-bl cells, while cytotoxicity was measured using an MTT assay. Antiviral activity was determined through a luciferase-based assay, and integration inhibition was evaluated using integrase activity assays and Alu-gag nested PCR. The extract demonstrated potent inhibition of resistant mutants, with low IC₅₀ values (0.02971–0.1652 μg/mL), and showed minimal cytotoxicity (CC₅₀ = 300 μg/mL), maintaining over 80% cell viability. It inhibited integrase activity by 67%, specifically targeting the strand-transfer step, and significantly reduced integrated viral DNA. Molecular docking of 14 compounds identified coumarin derivatives as key bioactive metabolites, exhibiting mutation-tolerant binding within the integrase catalytic pocket. Overall, these findings highlight PO4PR2 as a promising source of compounds for developing new therapies targeting drug-resistant HIV-1 integrase. Full article