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, ß-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.

Health risk assessment of triazole fungicides around a pesticide factory in China.

The environmental pollution and health effects caused by pesticide production have consistently garnered considerable research interest. In the present study, the concentrations of five triazole fungicides (TFs) in air, indoor dust, and diet were monitored around a pesticide factory in eastern China from November 2020 to May 2021. The levels of five TFs in each sample were determined via UPLC‒MS/MS. For a health risk assessment, the United States Environmental Protection Agency's deterministic method was applied. The findings revealed that the total concentrations of the five TFs around the monitoring area ranged from 0.29 to 5.85 ng/m3 in outdoor air, 287.4 to 9878.5 µg/kg in indoor dust, 0.0578 to 4.948 µg/kg in vegetables, and 0.447 to 3.00 µg/kg in rice. Notably, tebuconazole and hexaconazole had consistently high contributions over the years. For adults and children, the average daily doses (ADDs) were 1.32 × 10-5 and 2.69 × 10-5 mg/kg/day, respectively, in the monitoring area and 4.25 × 10-6 and 6.42 × 10-6 mg/kg/day, respectively, in the control area. In the control area, rice and vegetables were the primary media for exposure to TFs in children and adults, collectively accounting for more than 94% of the total TF exposure. Conversely, indoor dust is identified as the main medium of TF exposure in children residing near the pesticide factory, representing approximately 40% of the total exposure. The risks of noncarcinogenic effects on children and adults in the monitoring area were significantly greater than those in the control area, being approximately ten times greater for children, warranting increased attention. The carcinogenic risk to human health is relatively safe.

Pesticide impacts on insect pollinators: Current knowledge and future research challenges.

With the need to intensify agriculture to meet growing food demand, there has been significant rise in pesticide use to protect crops, but at different rates in different world regions. In 2016, the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services (IPBES) global assessment on pollinators, pollination and food production identified pesticides as one of the major drivers of pollinator decline.. This assessment highlighted that studies on the effects of pesticides on pollinating insects have been limited to only a few species, primarily from developed countries. Given the worldwide variation in the scale of intensive agricultural practices, pesticide application intensities are likely to vary regionally and consequently the associated risks for insect pollinators. We provide the first long-term, global analysis of inter-regional trends in the use of different classes of pesticide between 1995 and 2020 (FAOSTAT) and a review of literature since the IPBES pollination assessment (2016). All three pesticide classes use rates varied greatly with some countries seeing increased use by 3000 to 4000 % between 1995 and 2020, while for most countries, growth roughly doubled. We present forecast models to predict regional trends of different pesticides up to 2030. Use of all three pesticide classes is to increase in Africa and South America. Herbicide use is to increase in North America and Central Asia. Fungicide use is to increase across all Asian regions. In each of the respective regions, we also examined the number of studies since 2016 in relation to pesticide use trends over the past twenty-five years. Additionally, we present a comprehensive update on the status of knowledge on pesticide impacts on different pollinating insects from literature published during 2016-2022. Finally, we outline several research challenges and knowledge gaps with respect to pesticides and highlight some regional and international conservation efforts and initiatives that address pesticide reduction and/or elimination.

Induction of drug metabolizing enzyme and drug transporter expression by antifungal triazole pesticides in human HepaSH hepatocytes.

Triazole pesticides are widely used fungicides, to which humans are rather highly exposed. They are known to activate drug-sensing receptors regulating expression of hepatic drug metabolizing enzymes and drug transporters, thus suggesting that the hepatic drug detoxification system is modified by these agrochemicals. To investigate this hypothesis, the effects of 9 triazole fungicides towards expression of drug metabolizing enzymes and transporters were characterized in cultured human HepaSH cells, that are human hepatocytes deriving from chimeric humanized liver TK-NOG mice. Most of triazoles used at 10 µM were found to act as inducers of cytochrome P-450 (CYP) 1A1, CYP1A2, CYP2B6, CYP3A4 and UDP-glucuronosyltransferase 1A1 mRNA levels and of CYP3A4 protein; some triazoles also enhanced mRNA expression of the canalicular transporters P-glycoprotein/MDR1, multidrug resistance-associated protein 2 and breast cancer resistance protein. Triazoles however concomitantly inhibited CYP2B6 and CYP3A4 activities and thus appeared as dual regulators of these CYPs, being both inducers of their expression and inhibitors of their activity. The inducing effect however predominated, at least for bromuconazole, propiconazole and tebuconazole. Bromuconazole was moreover predicted to enhance CYP2B6 and CYP3A4 expression in humans exposed to this fungicide in a chronic, acute or occupational context. These data demonstrate that key-actors of the human hepatic detoxification system are impacted by triazole pesticides, which may have to be considered for the risk assessment of these agrochemicals. They additionally highlight that the use of human HepaSH cells as surrogates to primary human hepatocytes represents an attractive and promising way for studying hepatic effects of environmental chemicals.

Synthesis and antifungal activity of novel piperidinyl thiazole derivatives.

BACKGROUND: As an investigative program around the development of novel oomycetes fungicides, a systematic exploration of piperidinyl thiazole fungicides employing a bioisosteric replacement strategy was planned. RESULTS: A series of novel piperidinyl thiazole analogues were designed, synthesised, and evaluated as fungicides in vitro and in vivo against three oomycete fungal pathogens: Phytophthora infestans (P. infestans), Plasmopara viticola (P. viticola), and Pseudoperonospora cubensis (P. cubensis). Variation at four different regions of piperidinyl thiazole fungicides like Oxathiapiprolin and Fluoxapiprolin was considered. All variations delivered excellent in vitro and in vivo activity against P. infestans at extremely low concentrations. O-linked heterocyles, bicyclic piperidines, and sulfoximine-substituted aryls demonstrated the most robust transfer of in vitro activity into the greenhouse, while pyridones, acylated sulfoximines, and sultams showed less consistent results concerning the transfer. Specific compounds were selected for deeper evaluation of oomycetes activity. Several of them with representatives of all the series showed an excellent in vivo activity against P. viticola and P. cubensis. Among the most active compounds, P14 and P25 were further profiled in advanced studies and were found to have a robust curative potential. P14, P15, and P25 provided excellent control of P. infestans (in potato), P. viticola (in grapes), and P. cubensis (in cucumber) in field trials at dose rates of 20 and 30 g per hectare. CONCLUSIONS: A bioisosteric replacement strategy was successfully employed to identify novel piperidine thiazole derivatives with robust and commercially interesting control of oomycetes diseases in the greenhouse and under field conditions. A systematic exploration of various modifications of the piperidine thiazole class and their structure activity relationships was described. © 2024 Society of Chemical Industry.

Fungicide-albumin interactions: unraveling the complex relationship-a comprehensive review.

This review will give an insight into the interactions of serum albumins, which are proteins found in the blood, with fungicides. There are molecular interactions between several fungicides and two serum albumin proteins: human serum albumin (HSA) and bovine serum albumin (BSA). The main objective of this review is to through some light on the interactions of the fungicides with serum albumins and to highlight their toxicity level. The interactions of serum albumins with fungicides are complex and can be affected by the properties of the proteins themselves. This review provides valuable insight into the interactions between serum albumins and fungicides, which can help to know the efficacy and mechanism of fungicides and may help in designing new fungicides with low or no toxicity.

Exploring Endophytic Fungi as Natural Antagonists against Fungal Pathogens of Food Crops.

The yield and quality of cultivated food crops are frequently compromised by the prevalent threat from fungal pathogens that can cause widespread damage in both the pre-harvest and post-harvest stages. This paper investigates the challenges posed by fungal pathogens to the sustainability and yield of essential food crops, leading to significant economic and food security repercussions. The paper critiques the long-standing reliance on synthetic fungicides, emphasizing the environmental and health concerns arising from their widespread and occasionally inappropriate use. In response, the paper explores the potential of biological control agents, specifically endophytic fungi in advancing sustainable agricultural practices. Through their diverse symbiotic relationships with host plants, these fungi exhibit strong antagonistic capabilities against phytopathogenic fungi by producing various bioactive compounds and promoting plant growth. The review elaborates on the direct and indirect mechanisms of endophytic antagonism, such as antibiosis, mycoparasitism, induction of host resistance, and competition for resources, which collectively contribute to inhibiting pathogenic fungal growth. This paper consolidates the crucial role of endophytic fungi, i.e., Acremonium, Alternaria, Arthrinium, Aspergillus, Botryosphaeria, Chaetomium, Cladosporium, Cevidencealdinia, Epicoccum, Fusarium, Gliocladium, Muscodor, Nigrospora, Paecilomyces, Penicillium, Phomopsis, Pichia, Pochonia, Pythium, Ramichloridium, Rosellinia, Talaromyces, Trichoderma, Verticillium, Wickerhamomyces, and Xylaria, in biological control, supported by the evidence drawn from more than 200 research publications. The paper pays particular attention to Muscodor, Penicillium, and Trichoderma as prominent antagonists. It also emphasizes the need for future genetic-level research to enhance the application of endophytes in biocontrol strategies aiming to highlight the importance of endophytic fungi in facilitating the transition towards more sustainable and environmentally friendly agricultural systems.

Stereoselective toxicity: Investigating the adverse effects of benzovindiflupyr on Xenopus laevis tadpoles.

The novel chiral fungicide benzovindiflupyr exerts adverse effects on aquatic organisms; however, its toxic mechanism and stereoselectivity remain largely unknown. The current study aimed to investigate the enantioselective ecotoxicity mechanism of benzovindiflupyr in Xenopus laevis tadpoles using a 28-day exposure experiment. Results of the acute toxicity assessment indicated that (1S,4R)- and (1R,4S)-benzovindiflupyr exhibited high toxicity, with (1S,4R)- demonstrating approximately 75 times greater toxicity than (1R,4S)-. Compared to the latter, (1S,4R)-benzovindiflupyr significantly affected the growth, movement behavior, and oxidative stress of X. laevis tadpoles. The integration of metabolomics and transcriptomics data revealed that (1S,4R)-benzovindiflupyr disrupted the glycine, serine, and threonine metabolic pathways by modulating the activities of key enzymes. This dysregulation resulted in aberrant carbohydrate utilization, antioxidant pathways, and structural protein synthesis and degradation. Molecular docking confirmed that (1S,4R)-benzovindiflupyr exhibited superior docking activity with key enzymes, potentially contributing to its stereoselective toxicity. This study offers novel molecular perspectives on the enantioselective ecotoxicity mechanism of benzovindiflupyr toward aquatic organisms and highlights potential target proteins implicated in metabolic disorders.

Ultrasound assisted upper critical solution temperature type switchable deep eutectic solvent based liquid-liquid microextraction for the determination of triazole in water.

BACKGROUND: The use of pesticides to protect crops has long been an important measure to provide healthy and safe agricultural products, but excess pesticides flow into fields and rivers, causing environmental pollution. Earlier methods utilizing organic solvent liquid-liquid microextraction for pesticide residue detection were not environmentally friendly. Therefore, it is significant to find a greener and more convenient detection method to determine pesticide residues. RESULTS: A new method was established to detect three triazole fungicides (TFs), including myclobutanil, epoxiconazole and tebuconazole, in environmental water samples. And the determination was conducted using a high-performance liquid chromatography with the ultraviolet detector (HPLC-UV). The switchable deep eutectic solvent (SDES) can be reversibly switched between hydrophilic and hydrophobic states through temperature modulation. Additionally, the method exhibited excellent linearity for all target analytes within the concentration range of 10-2000 µg L-1, with satisfactory R2 values (≥0.9975). The limits of detection (LODs) ranged from 2.3 to 2.6 µg L-1, and the limits of quantification (LOQs) ranged from 7.8 to 8.7 µg L-1. The accuracy of the method was assessed through intra-day and inter-day precision tests, yielding relative standard deviations (RSDs) in the ranges of 2.8%-6.7% and 2.2%-7.5%, respectively. Density functional theory (DFT) results indicated that hydrogen bonding is a significant factor affecting the binding of DES with triazoles. Three different green assessment tools were used to prove that the SDES-HLLME method had good greenness and broad applicability. SIGNIFICANCE: This is a homogeneous liquid-liquid microextraction (HLLME) method based on the upper critical solution temperature (UCST) type switchable deep eutectic solvent program, which can complete the extraction within a few minutes without dispersant. In terms of pesticide detection, the analytical method is simple and more conducive to environmental protection.

Residential proximity to agricultural herbicide and fungicide applications and dust levels in homes of California children.

BACKGROUND: Few studies of the relationship between residential proximity to agricultural pesticide applications and pesticide levels in the home have incorporated crop location or wind direction. We evaluated the relationship between agricultural pesticide applications using the California Pesticide Use Reporting (CPUR) database and pesticide concentrations in carpet dust accounting for land use and wind direction. METHODS: We measured concentrations (ng/g) of seven herbicides and two fungicides in carpet dust samples from 578 California homes (2001-2007). We created three metrics by computing the density (kg/km2) of use of each pesticide reported in CPUR within 0.5-, 1-, 2-, and 4-km buffers around homes 180- and 365-days before sampling (CPUR metric). We apportioned applications to the crop area within the buffers (CROP-A metric) and weighted CPUR applications by the proportion of days that the home was within ±45° of the downwind direction (W-CPUR metric). We modeled natural-log concentrations (Tobit regression) and dust detections (logistic regression) adjusting for season/year, occupation, and home/garden use. RESULTS: Detections were >90 % for glyphosate, 2,4-D, and simazine. Detection rates and dust concentrations increased with increasing CPUR densities for all herbicides and one fungicide. Compared to homes without applications within 4 km, the highest tertile of 365-day glyphosate use was associated with ∼100 % higher concentrations (CPURT3>9.2kg/km2 %change = 110, 95 %CI = 55, 183; CROP-AT3>13.4kg/km2 %change = 144, 95 %CI = 81, 229; and W-CPURT3>2.1kg/km2 %change = 102, 95 %CI = 50, 171). The highest density tertiles of 2,4-D, simazine, and trifluralin were associated with 2- to 6-times higher concentrations, respectively; that was similar across metrics. Across all metrics, agricultural use of dacthal, dicamba, and iprodione were associated with 5- to 10-times higher odds of dust detections. Associations were unclear for 2-methyl-4-chlorophenoxyacetic acid and null for chlorothalonil. CONCLUSIONS: Agricultural herbicide and fungicide use was an important determinant of indoor contamination within 4 km of homes. Accounting for crops and wind direction did not substantially change these relationships.

Evaluation of the Antifungal and Biochemical Activities of Fungicides and Biological Agents against Ginseng Sclerotinia Root Rot Caused by Sclerotinia nivalis.

The objective of this study was to identify effective agents for the prevention and control of ginseng Sclerotinia root rot disease caused by Sclerotinia nivalis. The inhibitory effects of 16 chemical fungicides and 10 biocontrol agents (strains) on mycelial growth and sclerotium formation in S. nivalis were determined using a plate confrontation essay. The results showed that the best chemical agents for inhibiting the mycelial growth and sclerotium formation of S. nivalis were fluconazole and fludioxonil, while Bacillus amyloliquefaciens FS6 and B. subtilis (Kono) were the best biocontrol agents (strains). The results of field trials in 2022 and 2023 showed that the control effects of fluconazole and fludioxonil on ginseng Sclerotinia root rot disease were 90.60-98.16%, and those of the biocontrol agents B. amyloliquefaciens FS6 and B. subtilis (Kono) were 94.80-97.24%, respectively. Chemical agents produced abnormal and twisted mycelia, while the biocontrol agents increased mycelial branching, dilated the mycelium tip, and revealed an abnormal balloon. All of the fungicides decreased the ergosterol content, changed the cell membrane permeability, and increased the protein and nucleic acid permeability. These results suggest that these are potential agents for controlling ginseng Sclerotinia root rot disease, and their biochemical mechanisms of chemical and biocontrol of this disease were demonstrated.

Advancements in the nanodelivery of azole-based fungicides to control oil palm pathogenic fungi.

The cultivation of oil palms is of great importance in the global agricultural industry due to its role as a primary source of vegetable oil with a wide range of applications. However, the sustainability of this industry is threatened by the presence of pathogenic fungi, particularly Ganoderma spp., which cause detrimental oil palm disease known as basal stem rot (BSR). This unfavorable condition eventually leads to significant productivity losses in the harvest, with reported yield reductions of 50-80 % in severely affected plantations. Azole-based fungicides offer potential solutions to control BSR, but their efficacy is hampered by limited solubility, penetration, distribution, and bioavailability. Recent advances in nanotechnology have paved the way for the development of nanosized delivery systems. These systems enable effective fungicide delivery to target pathogens and enhance the bioavailability of azole fungicides while minimising environmental and human health risks. In field trials, the application of azole-based nanofungicides resulted in up to 75 % reduction in disease incidence compared to conventional fungicide treatments. These innovations offer opportunities for the development of sustainable agricultural practices. This review highlights the importance of oil palm cultivation concerning the ongoing challenges posed by pathogenic fungi and examines the potential of azole-based fungicides for disease control. It also reviews recent advances in nanotechnology for fungicide delivery, explores the mechanisms behind these nanodelivery systems, and emphasises the opportunities and challenges associated with azole-based nanofungicides. Hence, this review provides valuable insights for future nanofungicide development in effective oil palm disease control.

Design, Synthesis, Antifungal Evaluation, and Three-Dimensional Quantitative Structure-Activity Relationship of Novel 5-Sulfonyl-1,3,4-thiadiazole Flavonoids.

Plant pathogenic fungi frequently disrupt the normal physiological and biochemical functions of plants, leading to diseases, compromising plant health, and ultimately reducing crop yield. This study aimed to address this challenge by identifying antifungal agents with innovative structures and novel mechanisms of action. We designed and synthesized a series of flavonoid derivatives substituted with 5-sulfonyl-1,3,4-thiadiazole and evaluated their antifungal activity against five phytopathogenic fungi. Most flavonoid derivatives demonstrated excellent antifungal activity against Botrytis cinerea (B. cinerea), Alternaria solani (A. solani), Rhizoctorzia solani (R. solani), Fusarium graminearum (F. graminearum), and Colletotrichum orbiculare (C. orbiculare). Specifically, the EC50 values of 38 target compounds against R. solani were below 4 µg/mL, among which the compounds C13 (EC50 = 0.49 µg/mL), C15 (EC50 = 0.37 µg/mL), and C19 (EC50 = 0.37 µg/mL) had the most prominent antifungal activity, superior to that of the control drug carbendazim (EC50 = 0.52 µg/mL). Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images of the cellular ultrastructures of R. solani mycelia and cells after treatment with the compound C19 revealed sprawling growth of hyphae, a distorted outline of their cell walls, and reduced mitochondrial numbers. Studying the 3D-QSAR between the molecular structure and antifungal activity of 5-sulfonyl-1,3,4-thiadiazole-substituted flavonoid derivatives could significantly improve conventional drug molecular design pathways and facilitate the development of novel antifungal leads.

Direct and indirect effects of chemical pollution: Fungicides alter growth, feeding, and pigmentation of the freshwater detritivore Asellus aquaticus.

Anthropogenic chemical pollutants, such as fungicides, pose significant threats to natural ecosystems. Although the direct impacts of numerous chemicals are well-documented in simple environmental contexts, their indirect impacts are poorly understood. This study used two individual level laboratory experiments to assess direct and indirect effects of fungicides on the isopod Asellus aquaticus, a keystone detritivore in freshwater systems. First, a range-finding assay on three widely used fungicides (Fluazinam, Tebuconazole, Urea) showed that Tebuconazole had the strongest concentration-dependent negative effects on A. aquaticus growth and food consumption. Second, a factorial experiment using Tebuconazole assessed its direct and diet-mediated effects and showed that Tebuconazole reduced growth, feeding, and pigmentation through both pathways. The results indicate that assessing only direct impacts of toxic chemicals could overlook critical interactions that are relevant in natural systems, such as those associated with diet. Our study highlights the importance of considering both direct and indirect effects in environmental toxicology to better understand the full impacts of chemical pollutants in nature.

Recent innovations in crop protection research.

As the world's population continues to grow and demand for food increases, the agricultural industry faces the challenge of producing higher yields while ensuring the safety and quality of harvests, operators, and consumers. The emergence of resistance, pest shifts, and stricter regulatory requirements also urgently calls for further advances in crop protection and the discovery of new innovative products for sustainable crop protection. This study reviews recent highlights in innovation as presented at the 15th IUPAC International Congress of Crop Protection Chemistry held in New Delhi, in 2023. The following new products are discussed: the insecticides Indazapyroxamet, Dimpropyridaz and Fenmezoditiaz, the fungicides Mefentrifluconazole and Pyridachlomethyl, the nematicide Cyclobutrifluram, the herbicides Rimisoxafen, Dimesulfazet, and Epyrifenacil as well as the abiotic stress management product Anisiflupurin. In addition, the latest innovative research areas and discovery highlights in all areas of crop protection will be presented, including insecticidal alkyl sulfones and 1,3,4-trisubstituted pyrazoles, fungicidal picolinamides, herbicidal ketoenols, and trifluoromethylpyrazoles, as well as the latest advances in crop enhancement and green pest control research. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Imazalil and prochloraz toxicokinetics in fish probed by a physiologically based kinetic (PBK) model.

Azole fungicides are highly suspected endocrine disruptors (EDs) and are frequently detected in surface water. Among them, there are prochloraz (PCZ), a commonly used  molecule for ED studies, and imazalil (IMZ), a highly suspected ED. Little is known about their toxicokinetic (TK) behavior in fish. Hence, research suggested that an improved risk assessment could be achieved by gaining insight into their TK behavior. The aim of this study is to understand and model the TK of both substances in different fish species, irrespective of the scheme of exposure. TK data from the literature were retrieved including different modes of exposure (per os and waterborne). In addition, two experiments on zebrafish exposed to either IMZ or PCZ were performed to address the lack of in vivo TK data. A physiologically based kinetic (PBK) model applied to IMZ and PCZ was developed, capable of modeling different exposure scenarios. The parameters of the PBK model were simultaneously calibrated on datasets reporting internal concentration in several organs in three fish species (original and literature datasets) by Bayesian methods (Monte Carlo Markov Chain). Model predictions were then compared to other experimental data (i.e., excluded from the calibration step) to assess the predictive performance of the model. The results strongly suggest that PCZ and IMZ are actively transported across the gills, resulting in a small fraction being effectively absorbed by the fish. The model's results also confirm that both molecules are extensively metabolized by the liver into mainly glucuronate conjugates. Overall, the model performances were satisfying, predicting internal concentrations in several key organs. On average, 90% of experimental data were predicted within a two-fold range. The PBK model allows the understanding of IMZ and PCZ kinetics profiles by accurately predicting internal concentrations in three different fish species regardless of the exposure scenario. This enables a proper understanding of the mechanism of action of EDs at the molecular initiating event (MIE) by predicting bioaccumulation in target organs, thus linking this MIE to a possible adverse outcome.

Visual detection of fungicide resistance by combining RPA and CRISPR/Cas12a in peach Brown rot fungus Monilinia fructicola.

BACKGROUND: Peach brown rot caused by Monilinia fructicola severely affects the quality and yield of peach, resulting in large economic losses worldwide. Methyl benzimidazole carbamate (MBC) fungicides and sterol demethylation inhibitor (DMI) fungicides are among the most applied chemical classes used to control the disease but resistance in the target pathogen has made them risky choices. Timely monitoring of resistance to these fungicides in orchards could prevent control failure in practice. RESULTS: In the current study, we developed methods based on recombinase polymerase amplification (RPA) and CRISPR/Cas12a systems to detect MBC and DMI resistance based on the E198A mutation in the ß-tubulin (MfTub2) gene and the presence of the Mona element in the upstream region of the MfCYP51, respectively. For MBC resistance, RPA primers were designed that artificially incorporated PAM sites to facilitate the CRISPR/Cas12a reaction. Subsequently, specific tcrRNAs were designed based on the E198A mutation site. For the detection of the Mona element, we designed RPA primers M-DMI-F2/M-DMI-R1 that in combination with crRNA1 detected 'Mona' and distinguished resistant from sensitive strains. CONCLUSION: Both methods exhibited high sensitivity and specificity, requiring only a simple isothermal device to obtain results within 1 h at 37 °C. The FQ-reporter enabled visualization with a handheld UV or white light flashlight. This method was successfully used with purified DNA from lab cultures and crude DNA from symptomatic fruit tissue, highlighting its potential for on-site detection of resistant strains in orchards. © 2024 Society of Chemical Industry.

Population Structure of Phytophthora infestans in Israel Changes Frequently Due to the Import of Asymptomatic Late Blight-Infected Potato Seed Tubers from Europe.

Late blight, caused by the oomycete Phytophthora infestans, is a devastating disease of potato worldwide. In Israel, potatoes are grown twice a year, in autumn and spring, with late blight causing extensive damage in both seasons. While tuber seeds for the autumn planting are produced locally, seed tubers for the spring planting are imported from Europe due to dormancy of local tubers. Here, we demonstrate that seed tubers imported from Europe for the spring season carry asymptomatic infection with EU genotypes of P. infestans, which alters the population structure of the pathogen each spring. The proportion of imported tubers carrying asymptomatic infections ranged between 1.2 and 3.75%, varying by year and cultivar. Asymptomatic tubers produced late blight-infected sprouts about one month after planting. The sporangia produced on these sprouts served as primary inoculum, causing intensive foliage attacks on neighboring plants. When sprout-infected plants were uprooted and the mother tuber was washed, sliced, and placed in moistened dishes at 18 °C, profuse sporulation of P. infestans developed on the slices' surfaces within 1-2 days. The dominant genotype of P. infestans in the autumn season in Israel is 23A1, but genotypes in the following spring season changed to include 13A2 or 36A2. Surprisingly, genotype 43A1, which might be resistant to CAA and OSBPI fungicides and appeared in Europe in 2022, emerged in Israel in spring 2024. The immigrating genotypes do not persist in the country, allowing 23A1 to regain predominance in the following autumn. Long-term monitoring data suggest that the population structure of P. infestans changes yearly but temporarily due to the import of new genotypes from Europe.

Synthetic and Natural Antifungal Substances in Cereal Grain Protection: A Review of Bright and Dark Sides.

Molds pose a severe challenge to agriculture because they cause very large crop losses. For this reason, synthetic fungicides have been used for a long time. Without adequate protection against pests and various pathogens, crop losses could be as high as 30-40%. However, concerns mainly about the environmental impact of synthetic antifungals and human health risk have prompted a search for natural alternatives. But do natural remedies only have advantages? This article reviews the current state of knowledge on the use of antifungal substances in agriculture to protect seeds against phytopathogens. The advantages and disadvantages of using both synthetic and natural fungicides to protect cereal grains were discussed, indicating specific examples and mechanisms of action. The possibilities of an integrated control approach, combining cultural, biological, and chemical methods are described, constituting a holistic strategy for sustainable mold management in the grain industry.

Curse or blessing: Growth- and laccase-modulating properties of polyphenols and their oxidized derivatives on Botrytis cinerea.

Infection of grapevines with the grey mold pathogen Botrytis cinerea results in severe problems for winemakers worldwide. Browning of wine is caused by the laccase-mediated oxidation of polyphenols. In the last decades, Botrytis management has become increasingly difficult due to the rising number of resistances and the genetic variety of Botrytis strains. During the search for sustainable fungicides, polyphenols showed great potential to inhibit fungal growth. The present study revealed two important aspects regarding the effects of grape-specific polyphenols and their polymerized oxidation products on Botrytis wild strains. On the one hand, laccase-mediated oxidized polyphenols, which resemble the products found in infected grapes, showed the same potential for inhibition of growth and laccase activity, but differed from their native forms. On the other hand, the impact of phenolic compounds on mycelial growth is not correlated to the effect on laccase activity. Instead, mycelial growth and relative specific laccase activity appear to be modulated independently. All phenolic compounds showed not only inhibitory but also inductive effects on fungal growth and/or laccase activity, an observation which is reported for the first time. The simultaneous inhibition of growth and laccase activity demonstrated may serve as a basis for the development of a natural botryticide. Yet, the results showed considerable differences between genetically distinguishable strains, impeding the use of a specific phenolic compound against the genetic variety of wild strains. The present findings might have important implications for future understanding of Botrytis cinerea infections and sustainable Botrytis management including the role of polyphenols.