Synthetic Fungicides Research Articles

Efficacy of using grape cane extracts against <i>Plasmopara viticola</i> under field conditions and their impact on the composition of berries and musts of <i>Vitis vinifera</i> L. cv. Riesling

Grapevine downy mildew caused by Plasmopara viticola is a deleterious vine disease currently controlled using synthetic and copper-based fungicides. Environmental concerns surrounding such fungicides necessitate sustainable alternatives like grape cane extracts. However, studies into their efficacy and effects on the product remain limited. This study aimed to assess the open-field efficacy of a novel grape cane extract formulation, alone and combined with a copper agent, against downy mildew, and to examine its impact on berry and must composition. The results were compared to standard fungicides frequently used in integrated and organic viticulture. In 2022, the grape cane extract formulation reduced downy mildew severity by 78 % on berry clusters, comparable to the maximum acceptable dose of copper for organic farming in Germany (3 kg/ha/a). Combining the grape cane extract with the copper agent was more effective than the copper agent alone, but not more effective than the grape cane extract alone. In 2023, disease development was marginal, despite artificial inoculation. Compositional differences detectable by FTIR were insignificant between musts derived from all treatments, increased copper levels were observed whenever copper had been applied. The phenolic content in freeze-dried, de-seeded berries of 2022 was significantly lower after copper (14.6 mg/g dry weight DW) and grape cane extract treatments (6.6 mg/g DW) when compared to the control (36.1 mg/g DW), with discrepancies in stilbenoid and flavan-3-ol levels in particular, as analysed by HPLC-DAD. However, these differences were not confirmed in 2023, with results showing no correlations between different treatments and phenolic contents. These findings suggest that applications of the phenolic grape cane extract do not lead to artificially altered levels or relevant residues of the active constituents; i.e., the phenolic compounds in the product. In brief, grape cane extracts might represent a promising natural alternative to controlling grapevine downy mildew, particularly in organic viticulture, which, to date, heavily relies on (eco-)toxic copper-pesticides. Further multi-year studies including wine analyses are warranted to follow the entire flow towards the final product.

A comprehensive review of integrated management strategies for damping-off disease in chili

Damping-off disease in chili (Capsicum annum L.) cultivation is a significant global issue, severely affecting seeds, seedlings, and young plants, regardless of the location of cultivation, whether in greenhouses or open fields. Despite chili being a widely popular vegetable used in various cuisines globally, farmers face challenges in meeting the growing demand due to the extensive damage caused by this disease, ranging from 20 to 85%. The shelf life and quality of mature pods are also severely affected. Damping-off disease is mainly caused by soil-borne fungus from the Pythium species, with additional contributions from Phytophthora, Fusarium, and Rhizoctonia species. These pathogens’ adaptability to diverse environmental conditions and resistance to synthetic fungicides make controlling damping-off on a commercial scale challenging. However, integrated disease management has shown promising results as a remedial approach. In this review, we discuss the current state of chili diseases, the nature of the pathogens causing damping-off, the epidemiology of the disease, and various control mechanisms. In this review, we broadly discuss the current state of chili diseases, the nature of the pathogens causing damping-off, the epidemiology of the disease, and various control mechanisms. Furthermore, we highlight the importance and efficacy of integrated disease management techniques, along with future prospects in unexplored areas, such as host–pathogen interaction and sustainable disease control measures. The information in this review aims to assist chili growers in understanding the epidemiology and management of damping-off in chili cultivation.

Exploring the antifungal activity of clove oil (Syzygium aromaticum) against wilt disease caused by Fusarium equiseti in bitter gourd (Momordica charantia L.)

This study investigates the isolation and characterization of Fusarium equiseti, a fungal pathogen causing wilt in bitter gourd. It explores the antifungal activity of essential oils, particularly clove oil, as a natural management approach. The pathogen was isolated from wilt-infected bitter gourd stems and it was confirmed as F. equiseti morphologically by the presence of three-septate falcate macroconidia and globose microconidia and also confirmed molecularly by PCR amplification of the ITS region (PP501044.1). Among the tested essential oils, clove oil demonstrated the highest mycelial growth inhibition (100%) at a concentration of 0.5%, followed by peppermint, wintergreen and tea tree oils. In comparison, neem oil exhibited the least inhibition (15.54%). GC-MS analysis of clove oil revealed 38 compounds, where eugenol is the predominant compound that played a crucial role in antifungal activity. Additionally, metabolic analysis revealed several enriched pathways in F. equiseti during its interaction with clove oil, particularly those involved in lipid metabolism and energy production. Pot culture experiments confirmed the efficacy of clove oil in reducing disease severity and incidence in bitter gourd, achieving a 66.99 % reduction when combined with soil application.Furthermore, clove oil-treated plants showed increased activity of defense-related enzymes, including peroxidase (PO), polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL), suggesting the induction of systemic resistance. This study highlights the potential of clove oil as an eco-friendly alternative to synthetic fungicides for managing F. exquisite-induced wilt in bitter gourd. Future research should focus on optimizing clove oil application methods and understanding its interactions with fungal pathogens at the molecular level to enhance its efficacy in sustainable disease management.

Exploring the Potential of Bacterial Endophytes in Plant Disease Management.

Endophytic bacteria live in the internal tissues of plants, forming symbiotic, mutualistic, commensalistic and trophobiotic relationships. Some are spread via seeds after sprouting from the rhizosphere or phyllosphere. These bacteria capable of promoting plant growth and impart biotic stress by synthesing plant growth hormones, ACC deaminase, organic acids and siderophore. Endophytes aid in phytoremediation by removing soil contaminants and boosting soil fertility via phosphate solubilization and nitrogen fixation. The endophytic microbes are becoming increasingly popular in biotechnological applications which supports sustainable growth of non-food crops for biomass and biofuel. They offer valuable natural materials which is used in medicine, agriculture and industry. Bacterial endophytes are endowed with the enormous potential in the biological treatment of plant pathogens and considered as the superior alternative to synthetic fungicides. The review emphasizes benefits of bacterial endophytes in promoting plant growth and prospects of agricultural applications viz., increasing crop yield under biotic stress condition and their mode of action towards plant diseases. It also summarises the diverse and vital role of endophytes in agroecosystems as well as insights for sustainable agriculture and crop resilience.

Amplification-free detection of Ascochyta blight in chickpea using a simple molecular beacon assay

Ascochyta blight is a major biotic stress that limits chickpea production globally. Fungicide application remains one of the effective control measures for the endemic spread. Due to the serious threat that synthetic fungicides pose to crop quality, early diagnosis of the pathogen is imperative. Whilst there have previously been several conventional lab-based diagnostic methods developed for early detection of Ascochyta rabiei, they require long assay times, specialised equipment and facilities, and trained personnel to process the samples. To overcome this challenge, a rapid amplification-free detection assay using a molecular beacon probe was developed. The method consists of a simple assembly assay that accurately detects pathogen within 30 min. The developed assay is species-specific and has a similar sensitivity level as conventional amplification-based methods. Although it is still a lab-based technique, considering the simplicity of the assay, it has a great potential to be developed further as a reliable in-field diagnostic device for early detection and quantification of fungal pathogen spores.

New Trichoderma Strains Suppress Blue Mold in Oranges by Damaging the Cell Membrane of Penicillium italicum and Enhancing Both Enzymatic and Non-Enzymatic Defense Mechanisms in Orange Fruits

Blue mold disease, caused by Penicillium italicum (P. italicum), presents a significant challenge to orange fruits (Citrus sinensis L.) and other citrus crops globally. Biological control, particularly Trichoderma species, offers a promising alternative to synthetic fungicides. Therefore, this study aimed to isolate, identify, and evaluate the antagonistic activities of two Trichoderma isolates against P. italicum. These isolates were molecularly identified and assigned accession numbers PP002254 and PP002272, respectively. Both isolates demonstrated significant antifungal activity in dual culture assays. Moreover, the culture filtrates (CFs) of Trichoderma longibrachiatum PP002254 and Trichoderma harzianum PP002272 suppressed the mycelial growth of P. italicum by 77.22% and 71.66%, respectively. Additionally, CFs reduced the severity of blue mold on orange fruits by 26.85% and 53.81%, compared to 100% in the control group. Scanning electron microscopy revealed that treated P. italicum hyphae were shrunken and disfigured. Enzyme activities (catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase) in treated oranges increased, along with total soluble phenolics and flavonoids. Conversely, malondialdehyde (MDA) levels decreased in treated fruits. These findings suggest that T. longibrachiatum PP002254 and T. harzianum PP002272 could be effective biocontrol agents for managing blue mold and other citrus postharvest diseases.

Assessing variability among culturable phylloplane basidiomycetous yeasts from Italian agroecosystems

This study analysed basidiomycetous yeasts isolated from the phylloplane of crops and spontaneous plants in Italian agroecosystems. A total of 25 species belonging to 17 genera were recognized by analysing 83 isolates from vineyards and orchards, that are not treated with synthetic fungicides, and adjacent natural areas. Rhodotorula graminis and Filobasidium magnum were the most frequent species but 13 others were represented by a single isolate (e.g., Buckleyzyma salicina, Pseudozyma prolifica, and Moniliella megachiliensis). Preliminary analysis of (GTG)5-PCR fingerprinting revealed high genetic intraspecific heterogeneity. All isolates were characterized by their production of extracellular hydrolytic enzymes and their sensitivity to six commercial fungicides used in Italy. The isolates displayed great variability in these phenotypic traits, which play an important role in the survival of yeast populations in agroecosystems. Most of them exhibited lipolytic, proteolytic, β-glucosidase and pectinolytic activities, but only three (F. magnum, Kwoniella mangroviensis and Ps. prolifica) also had cellulolytic and amylolytic activity. Most isolates were sensitive to four fungicides, and one R. graminis isolate was resistant to all six. This heterogeneity was not related to the geographical origin of the isolates. The lack of selective factors (i.e. pesticide treatments) in the sampling fields and the presence of adjacent natural areas may have favored the maintenance of an elevated level of strain diversity. This study provides new information on phylloplane basidiomycetous yeasts in agroecosystems and opens the way to further investigations into the impact of agricultural practices on the microbial diversity of these natural habitats.

Assessing Biofungicides and Host Resistance against Rhizoctonia Large Patch in Zoysiagrass.

Rhizoctonia large patch (Rhizoctonia solani AG2-2 LP) significantly reduces turfgrass quality, aesthetics, and playability. Synthetic fungicides are commonly used for managing this disease, but they present high costs, potential for fungicide resistance, and environmental concerns. We conducted in vitro assays to test the effectiveness of three biofungicides, seven synthetic fungicides, and ten combinations against R. solani. We then assessed seven spray programs that included Bacillus subtilis QST713 and propiconazole, either alone or tank-mixed, on zoysiagrass 'El Toro' in a growth chamber and in field trials. Biofungicide B. subtilis QST713 reduced pathogen growth by up to 100% in vitro. B. subtilis QST713 alone or combined with synthetic fungicides and/or in rotation was as effective as the standalone synthetic fungicide, reducing disease severity and AUDPC by 81 and 77% (growth chamber) and by 71 and 52% (field), respectively, while maintaining acceptable turfgrass quality. Additionally, we screened zoysiagrass genotypes and advanced breeding lines against three R. solani isolates in growth chamber studies. Five genotypes and two breeding lines demonstrated resistance to Rhizoctonia large patch across isolates, highlighting their potential for developing disease-resistant cultivars. Our findings suggest that integrating biofungicides, resistant cultivars with chemical controls offer sustainable and effective strategies for managing Rhizoctonia large patch.

Potentiality of Formulated Pseudomonas Fluorescens and Bacillus Subtilis for the Management of Sheath Blight Disease of Rice Under Field Condition

Rice production is severely affected by sheath blight disease in Bangladesh which is caused by a fungus Rhizoctonia solani that is still merely managed by synthetic chemical fungicides. In this study, different isolates of Pseudomonas fluorescens and Bacillus subtilis were evaluated to assess their inhibitory effects against R. solani both in- vitro and field condition following dual culture method in order to formulate technique for the management of sheath blight of rice. Among all the isolates, P. fluorescens (Pf- 7 and Pf- 8) and B. subtilis (Bs-17 and Bs-21) showed the highest in –vitro mycelial growth suppression of the pathogen. Based on the in-vitro mycelial growth suppression performance, fourteen treatments combinations, either single or in consortia of antagonistic bacteria in talc-based formulations were applied by root dipping. All the formulated bacterial isolates showed significant reduction in disease and severity compared to untreated control and chemical fungicide. At 90 DAT, both the disease incidence and severity were found lowest in T11 (31.67 % and 37.33 % respectively) where the seedlings were treated for 24 hours with consortium of Pf-7 + Pf- 8 formulated in talc followed by in T12 (32.00 % and 38.00 %) where the seedlings were treated for 24 hours with consortium of Bs-17 + Bs-21 formulated in talc respectively. Vegetative and yield parameters were also significantly increased in both T11 and T12. The highest yield as well as Benefit Cost Ratio (BCR) were found in T11 (2.50 kg/ m2 and 2.20) followed by T12 (2.49 kg/m2 and 2.18). Thus, the consortia of bacterial isolates (Pseudomonas fluorescens and Bacillus subtilis) with seedling dipping time for 24 hour represents a promising alternative to chemical fungicides for the management of sheath blight of rice in field condition. J Bangladesh Agril Univ 22(3): 342-351, 2024

Sustainable management of blister and grey blight diseases of tea using antagonistic and plant growth promoting microbes in Western Ghats of India

The blister and grey blights are the devastating leaf diseases in tea. The severity of blister and grey blight diseases in tea ranged from 22% to 68% and 11.41%–57.02%, respectively in a survey conducted in Southern India during 2022–2023. The morphological and molecular identifications of pathogens revealed that Exobasidium vexans and Neopestalotiopsis clavispora were responsible for causing blister and grey blights on tea in southern India. The conventional management strategy with foliar spray of synthetic fungicides result in fungicidal residue in tea causes environmental pollution and health hazards to human. To overcome this the current study aims to manage the blister and grey blight diseases of tea through the microbial biocontrol agents. The bacterial biocontrol agents viz., Bacillus subtilis (BBV57), Bacillus amyloliquefaciens (TNAU), Pseudomonas fluorescens (Pf1) and the fungal antagonist, Trichoderma asperellum (TV 1) were evaluated for their efficacy against the both blight diseases. All the bio agents have significantly reduced the basidiospore germination and the mycelial growth of E. vexans and N. clavispora, respectively. Among the bio agents, B. amyloliquefaciens registered 88.36% reduction in the germination of E. vexans basidiospore and 85.56% of mycelial growth reduction of N. clavispora in vitro (P < 0.05). Under field conditions the combined soil and foliar application of bacterial antagonists viz., B. subtilis, B. amyloliquefaciens and P. fluorescens each with 1 ˟ 1011 CFU/ml @ 5% concentration at 7 days interval significantly reduced the blister blight incidence with 53.53% & 51.53% reduction over control and 51.53% & 43.65% of reduction over initial during first and second year, respectively. The combined application also reduced the grey blight incidence with 53.53% & 52.00% reduction over control and 56.81% & 47.65% of reduction over initial during first and second year, respectively. This treatments with bioagents were almost comparable with synthetic treatments, such as copper oxy chloride (0.25%) and hexaconazole (0.25%) with 64.63% and 64.76% reduction of blister and grey blight reduction in first year; 59.74%, 51.11% reduction in second year over control (P < 0.05), respectively. Thus, the application of B. subtilis, B. amyloliquefaciens and P. fluorescens can be recommended for the sustainable management of tea blister and grey blight diseases in tea gardens.

NaCl inhibits Geotrichum citri-aurantii growth by inducing lipid droplets accumulation and enhances the biocontrol efficacy of Kloeckera apiculata 34-9

Decays resulting from infections by Geotrichum citri-aurantii cause a significant reduction in citrus postharvest quality and marketable yield. In the present work, 0.8 mol · L-1 NaCl of Generally Recognized as Safe (GRAS) salts were identified to have complete inhibition activity to G. citri-aurantii in vitro and in vivo. An inhibitory mechanism study showed that 0.1 mol · L-1 NaCl changed the expression of six genes (Gci002087, Gci003354, Gci000013, Gci000394, Gci003505 and Gci004433) in target of rapamycin (TOR) pathway, thus inducing a large number of lipid droplets (LDs) accumulation with TEM observations and staining. In vivo, 0.1 mol · L-1 NaCl could raise up the activities of CAT, PAL, PPO and cellulase which hence to strengthen the ability of resistance tolerance of citrus. In addition, NaCl not only accelerated the growth of the biocontrol yeast Kloeckera apiculata strain 34-9, but also increased the expression level of adhesin-encoding genes and biofilm-formation encoding gene (34-9 - 3691). Finally, the storage experiments in 3 months demonstrated that the combination of NaCl and the yeast 34-9 could keep fresh, reduce decay and prolong its shelf-life, thus implying that the combination of biocontrol strain and GRAS salts has been explored as a promising alternative to synthetic fungicides.

Investigation of Antimicrobial Compounds Produced by Endolichenic Fungi in Different Culture Media.

Continuous use of synthetic fungicides has led to explosive emergence of fungicide-resistant microbes. Therefore, there are urgent needs for environmentally friendly antimicrobial agents with novel modes of action. This study investigated endolichenic fungi (ELF) as a source of antimicrobial compounds against various plant pathogens. We utilized an One Strain MAny Compounds (OSMAC) approach to enhance the chemical diversity of fourteen ELF. This involved cultivation of ELF in four growth media and subsequently assessing antimicrobial activities of culture extracts. Nearly half of the culture extracts exhibited antimicrobial activity against a Gram-positive bacterium, but showed minimal activity against Gram-negative bacteria tested. Notably, culture extracts from two ELF, Chaetomium globosum and Nodulisporium sp., demonstrated significant inhibitory effects against plant pathogenic fungi. LC-MS/MS-based metabolome profiling confirmed the presence of known bioactive compounds like cyclic dipeptides and chaetoglobosins. These findings highlight the effectiveness of combining OSMAC and metabolomics for identifying antimicrobial agents for agricultural use.

Characterization and biocontrol potential of Wickerhamomyces subpelliculosus yeasts isolated from dates: Volatile compounds-mediated antifungal activity against mycotoxigenic Penicillium strains

Seven yeast strains were isolated from Tunisian dates. The strains were identified by sequence analysis of the D1/D2 domain of the nuclear large subunit (LSU) rRNA gene. Based on this all strains in the study were almost identical with that of the type strain of Wickerhamomyces subpelliculosus (CBS 5767) indicating that they belong to this species. All strains were characterized physiologically and biochemically. All strains grew in the presence of 50 % sucrose, 10 % sodium chloride and at 42 °C. The potential of these yeasts as biocontrol agent against mycotoxigenic Penicillium species inhabiting date, was evaluated. All yeast strains inhibited the growth of P. citrinum P10 and P. chrysogenum C17 previously isolated from dates, with inhibition percentages ranging between 43.6 % and 70.3 % on dual culture plate assays. Moreover, the volatile compounds (VCs) produced by these yeasts inhibited the mycelial growth rate and sporulation of both fungus strains, up to 76.5 and 100 %, respectively, on inverted culture plate assay. The VCs of W. subpelliculosus strains Y4 and Y24, which exhibit strong inhibitory activity against toxigenic Penicillium, were determined by head-space solid-phase microextraction (HS-SPME) combined with gas chromatography coupled with mass spectrometry (GC–MS) analysis. Results revealed significant levels of alcohols (27.36 % for Y4 and 23.35 % for Y24) and esters (66.19 % for Y4 and 75.82 % for Y24). Their significant bioactivity, along with the lack of reported adverse effects on consumer health or the environment, makes them a sustainable and effective alternative to synthetic fungicides for the biocontrol of mycotoxigenic Penicillium affecting stored dates.

Inhibition of Key Virulence Enzymes of Botrytis cinerea by Flavonoids from the Antarctic Plant Colobanthus quitensis.

This study investigates the antifungal efficacy of flavonoids derived from Colobanthus quitensis against key virulence-related enzymes implicated in the pathogenic mechanisms of Botrytis cinerea. The flavonoids swertiajaponin, schaftoside, vitexin, and saponarin significantly inhibited pectinase, cellulase, and laccase activity. Specifically, swertiajaponin showed mixed inhibition of pectinase and cellulase, characterized by high affinity (low inhibition constant -Ki-) for enzyme-substrate complexes. Schaftoside showed mixed inhibition of pectinase and competitive inhibition of laccase, effectively reducing enzymatic activity by competing directly with the substrate. In contrast, vitexin showed competitive inhibition of pectinase and non-competitive inhibition of laccase, suggesting it induces conformational changes within the enzyme. Finally, saponarin uniquely showed uncompetitive inhibition of laccase, stabilizing the enzyme-substrate complex and thereby markedly reducing catalytic turnover. Supported by kinetic parameters (maximum velocity -Vmax-, Michaelis constant -Km-, and Ki), these findings highlight the potential of flavonoids from C. quitensis as natural fungicides, offering a sustainable and eco-friendly alternative to synthetic fungicides for managing agricultural diseases.

Exploring marine compounds as potential biocontrol agents against powdery mildew for agricultural sustainability: a computer-based approach

Powdery mildew is a pervasive fungal disease causing significant economic losses globally. Continuous use of synthetic fungicides has led to environmental concerns and resistant fungal strains. This study explores marine-derived cephalostatins from the South African Natural Compounds Database as novel fungicidal agents for managing powdery mildew. Using molecular docking techniques, we investigated the interaction between selected cephalostatins and critical proteins involved in powdery mildew pathogenesis. Compounds were selected based on drug-likeness and bioactivity, adhering to Lipinski’s Rule of Five. Molecular interactions, binding affinities, and stability were analysed using AutoDock Vina, PyMOL, and Discovery Studio. Cephalostatin 17 exhibited the highest binding affinity (−10.4 kcal/mol), indicating strong potential for inhibiting fungal growth through significant hydrogen bonding and hydrophobic interactions. The study’s primary limitation is the reliance on computational predictions, necessitating experimental validation. Cephalostatin 17 stands out as a promising candidate for sustainable agricultural practices.

Are1-mediated nitrogen metabolism is associated with iron regulation in the mycoparasite Trichoderma atroviride

Trichoderma atroviride is a mycoparasitic fungus with antagonistic activity against fungal pathogens and is used as a pathogen control agent alternative to synthetic fungicides. Sensing nutrient availability in the environment and adjusting metabolism for optimal growth, development and reproduction is essential for adaptability and is relevant to its mycoparasitic activity. During mycoparasitism, secondary metabolites are produced to weaken the fungal prey and support the attack. Are1-like proteins act as major GATA-type transcription factors in the activation of genes subject to nitrogen catabolite repression. Since the quality and quantity of nitrogen has been proven particularly relevant in remodeling the biosynthesis of secondary metabolites in fungi, we decided to functionally characterize Are1, the ortholog of Aspergillus nidulans AreA, in T. atroviride. We show that the growth of the T. atroviride ∆are1 mutant is impaired in comparison to the wild type on several nitrogen sources. Deletion of are1 enhanced sensitivity to oxidative and cell-wall stressors and altered the mycoparasitic activity. We were able to identify for the first time a link between Are1 and iron homeostasis via a regulatory mechanism that does not appear to be strictly linked to the nitrogen source, but rather to an independent role of the transcription factor.

Selecting Bacillus strains antagonist to Erysiphe necator (Schw.) Burr. the causal agent of grape powdery mildew

ABSTRACT Grape powdery mildew (GPM) control is based on the preventive use of sulfur and curative application of synthetic fungicides, increasing the risk for producers’ health and environmental and fruit contamination. This work aimed to select Bacillus strains isolated from the Brazilian tropical semi-arid region that are effective antagonists to GPM. In an initial screening performed by spraying bacterial suspensions in detached grape leaves, six strains of Bacillus spp. showed disease symptom reduction higher than 70.0%. Two greenhouse experiments showed that the bacterial strains LCB03, LCB28, and LCB30 showed control efficiency >80%, statistically similar to a commercial formulation with Bacillus amyloliquefaciens QST713. 16s rDNA sequencing showed that strain LCB03 showed 100.0% homology to B. velezensis, while LCB28 has high homology to B. tequillensis (99.93%), and LCB30 had 99.71% homology with B. siamensis. As an average for both greenhouse experiments, weekly application of Bacillus sp LCB03, Bacillus sp LCB28, and Bacillus sp. LCB30 reduced the average incidence by around 50% and more than 80% for GPM severity.

Inhibitory Effect of Sorbus aucuparia Extracts on the Fusarium proliferatum and F. culmorum Growth and Mycotoxin Biosynthesis.

Fungal infections are among the most common diseases of crop plants. Various species of the Fusarium spp. are naturally prevalent and globally cause the qualitative and quantitative losses of farming commodities, mainly cereals, fruits, and vegetables. In addition, Fusarium spp. can synthesize toxic secondary metabolites-mycotoxins under high temperature and humidity conditions. Among the strategies against Fusarium spp. incidence and mycotoxins biosynthesis, the application of biological control, specifically natural plant extracts, has proved to be one of the solutions as an alternative to chemical treatments. Notably, rowanberries taken from Sorbus aucuparia are a rich source of phytochemicals, such as vitamins, carotenoids, flavonoids, and phenolic acids, as well as minerals, including iron, potassium, and magnesium, making them promising candidates for biological control strategies. The study aimed to investigate the effect of rowanberry extracts obtained by supercritical fluid extraction (SFE) under different conditions on the growth of Fusarium (F. culmorum and F. proliferatum) and mycotoxin biosynthesis. The results showed that various extracts had different effects on Fusarium growth as well as ergosterol content and mycotoxin biosynthesis. These findings suggest that rowanberry extracts obtained by the SFE method could be a natural alternative to synthetic fungicides for eradicating Fusarium pathogens in crops, particularly cereal grains. However, more research is necessary to evaluate their efficacy against other Fusarium species and in vivo applications.

Trichoderma harzianum TIND02 upregulates the expression of pathogenesis-related genes and enzymes and enhances gray blight resistance in tea

The gray blight incited by Pestalotiopsis and allied genera is a prevalent disease affecting tea cultivation, and managing it with Trichoderma spp. is an alternative to synthetic fungicides. Plants modify their arsenal system against pathogens when they are exposed to Trichoderma spp., which produces proteins and enzymes associated with pathogenesis. Understanding the expression pattern of defense-related markers will help in developing gray blight resistance tea cultivars. Thus, this study intended to induce resistance against gray blight in tea by Trichoderma harzianum TIND02. For this, a total of eight Trichoderma isolates originated from organic tea rhizospheres were characterized and evaluated for their efficacy. Dual culture test revealed isolate TIND02 as the most potential candidate with 74.6% inhibitory activity against gray blight pathogen Pseudopestalotiopsis theae. Molecular characterization based on ITS and tef-1 alpha genes confirmed isolate TIND02 as T. harzianum. Scanning electron microscopic study showed the mycoparasitic nature of T. harzianum TIND02 (TH-TIND02) to Ps. theae. The ethyl acetate extract of TH-TIND02 at 100 and 200 μg mL−1 showed potential inhibitory activity (>69.9%) against Ps. theae which confirmed the presence of higher volatile metabolites. Gas chromatography–Mass spectrometry study revealed that ethyl acetate extract of TH-TIND02 was composed of 21 major and minor volatile organic compounds with acetamide, 2, 2, 2-trifluoro-N, N-bis trimethyIsilyl–C (94.74%) as a major component. The isolate also produced chitinase, cellulase, β-1, 3 glucanase, and protease hydrolytic enzymes. Nursery experiments revealed that 2% and 5% doses (2 × 106 CFU mL−1) of TH-TIND02 significantly reduced respective 65.0% and 70.0% disease severity over control with improved plant growth. Besides, expressions of defense-related enzymes (chitinase, pHenolics, peroxidase, phenylalanine ammonia lyase, β-1, 3-glucanase, and polyphenol oxidase) and pathogenesis-related genes (chitinase and β-1, 3-glucanase) due to TH-TIND02 were determined. The secretion of defense-related enzymes was highly upregulated in plants applied with TH-TIND02 followed by Ps. theae inoculation compared to controls. The RT-qPCR analysis showed that the expression of both genes in co-inoculated plants was two-fold higher than in control after 21-day post incubation. These results suggest that TH-TIND02 application reduced gray blight severity by elevated enzyme activity and overexpressed pathogenesis-related genes in tea plants which offer for its eco-friendly and sustainable use as a bio-fungicide in tea gardens.

Interventions based on alternative and sustainable strategies for postharvest control of anthracnose and maintain quality in tropical fruits.

Colletotrichum spp. is a phytopathogen causing anthracnose in a variety of tropical fruits. Strategies used to control postharvest diseases in tropical fruits typically rely on the use of synthetic fungicides, which have stimulated the emergence of resistant pathogens. Safer alternative strategies to control anthracnose in tropical fruits have been described in the literature. This review presents and discusses the main innovative interventions concerning the application of sustainable alternative strategies in the postharvest control of pathogenic Colletotrichum species in tropical fruits, with a particular emphasis on the studies published in the last 5 years. The available studies have shown the use of various methods, including physical barriers, natural antimicrobials, and biological control with antagonistic microorganisms, to reduce anthracnose lesion severity and incidence in tropical fruits. The available literature showed high inhibitory activity in vitro, reduced anthracnose incidence and lesion diameter, and total disease inhibition in tropical fruits. Most studies focused on the inhibition of Colletotrichum gloeosporioides on avocado, papaya, and mango, as well as of Colletotrichum musae on banana; however, the inhibition of other Colletotrichum species was also demonstrated. The application of emerging sustainable alternative methods, including natural antimicrobial substances, also stimulated the induction of defense systems in tropical fruits, including enzymatic activity, such as polyphenol oxidase, peroxidase, and phenylalanine ammonia-lyase. The retrieved data helped to understand the current state of the research field and reveal new perspectives on developing efficient and sustainable intervention strategies to control pathogenic Colletotrichum species and anthracnose development in tropical fruits.