Fungicides Iprodione Research Articles

Life Cycle Assessment as a Decision-Making Tool for Photochemical Treatment of Iprodione Fungicide from Wastewater

Water contamination with various micropollutants is a serious environmental concern since this group of chemicals cannot always be removed efficiently with advanced treatment methods. Therefore, alternative chemical- and energy-intensive oxidation processes have been proposed for the removal of refractory and/or toxic chemicals. However, similar treatment performances might result in different environmental impacts. Environmental impacts can be determined by adopting a life cycle assessment methodology. In this context, lab-scale experimental data related to 100% iprodione (a hydantoin fungicide/nematicide selected as the model micropollutant at a concentration of 2 mg/L) removal from simulated tertiary treated urban wastewater (dissolved organic carbon content = 10 mg/L) with UV-C-activated persulfate treatment were studied in terms of environmental impacts generated during photochemical treatment through the application of a life cycle assessment procedure. Standard guidelines were followed in this procedure. Iprodione removal was achieved at varying persulfate concentrations and UV-C doses; however, an “optimum” treatment condition (0.03 mM persulfate, 0.5 W/L UV-C) was experimentally established for kinetically acceptable, 100% iprodione removal in distilled water and adopted to treat iprodione in simulated tertiary treated wastewater (total dissolved organic carbon of iprodione + tertiary wastewater = 11.2 mg/L). The study findings indicated that energy input was the major contributor to all the environmental impact categories, namely global warming, abiotic depletion (fossil and elements), acidification, eutrophication, freshwater aquatic ecotoxicity, human toxicity, ozone depletion, photochemical ozone creation, and terrestrial ecotoxicity potentials. According to the life cycle assessment results, a concentration of 21.42 mg/L persulfate and an electrical energy input of 1.787 kWh/m3 (Wh/L) UV-C light yielded the lowest undesired environmental impacts among the examined photochemical treatment conditions.

Physiological Quality of Treated and Stored Barley Seeds

Industrial seed treatment (IST) is used in some grain crops. However, the products used often impair the quality of seeds, especially insecticides. In the case of barley seeds, investigative studies on the use of IST are scarce. The present study aimed to evaluate the influence of IST with the fungicide (Iprodione) and insecticide (Thiamethoxan) products, mixed or not, on the physiological quality of barley seeds, before and after storage for two months. A completely randomized design was used, in a 4x2 factorial scheme, with four replications. The treatments consisted of four combinations of applications carried out via IST on barley seeds, of chemical pesticides (fungicide/insecticide), alone or in mixtures, as follows: {T1 – fungicide Iprodione (Rovral); T2 – Thiamethoxan insecticide (Cruizer); T3 – mixture of fungicide/insecticide products (Iprodione + Thiamethoxan); T4 – Control)}, submitted to storage for two months, with evaluations carried out at T0 (treated and analyzed seeds) and T1 (treated seeds stored for 60 days). The following tests were carried out: water content, germination, first count, accelerated aging, seedling length, seedling dry mass. It was concluded that the germination of barley seeds treated with the fungicide Iprodione (Rovral®) and the insecticide Thiamethoxan (Cruizer®), alone and in a mixture applied via IST, maintained the physiological quality of the seeds within the standards for commercialization until 60 days of storage. The use of the insecticide Thiamethoxan (Cruizer®) via IST showed greater damage to the physiology of barley seeds during storage (60 days), applied alone or in a mixture with the fungicide Iprodione (Rovral ®).

A new point mutation (D1158N) in histidine kinase Bos1 confers high-level resistance to fludioxonil in field gray mold disease

Gray mold, caused by the fungus Botrytis cinerea, is one of the most important plant diseases worldwide that is prone to developing resistance to fungicides. Currently, the phenylpyrrole fungicide fludioxonil exhibits excellent efficacy in the control of gray mold in China. In this study, we detected the fludioxonil resistance of gray mold disease in Shouguang City of Shandong Province, where we first found fludioxonil-resistant isolates of B. cinerea in 2014. A total of 87 single spore isolates of B. cinerea were obtained from cucumbers in greenhouse, and 3 of which could grow on PDA plates amended with 50 μg/mL fludioxonil that was defined as high-level resistance, with a resistance frequency of 3.4%. Furthermore, the 3 fludioxonil-resistant isolates also showed high-level resistance to the dicarboximide fungicides iprodione and procymidone. Sequencing comparison revealed that all the 3 fludioxonil-resistant isolates had a point mutation at codon 1158, GAC (Asp) → AAC (Asn) in the histidine kinase Bos1, which was proved to be the reason for fludioxonil resistance. In addition, the fludioxonil-resistant isolates possessed an impaired biological fitness compared to the sensitive isolates based on the results of mycelial growth, conidiation, virulence, and osmotic stress tolerance determination. Taken together, our results indicate that the high-level resistance to fludioxonil caused by the Bos1 point mutation (D1158N) has emerged in the field gray mold disease, and the resistance risk is relatively high, and fludioxonil should be used sparingly.

Metabolic Profiling and Comparative Proteomic Insight in Respect of Amidases during Iprodione Biodegradation.

The fungicide iprodione (IPR) (3-(3,5-dichlorophenyl) N-isopropyl-2,4-dioxoimidazolidine-1-carboxamide) is a highly toxic compound. Although IPR has been restricted, it is still being applied in many places around the world, constituting an environmental risk. The biodegradation of IPR is an attractive option for reducing its residues. In this study, we isolated thirteen IPR-tolerant bacteria from a biopurification system designed to treat pesticides. A study of biodegradation using different strains was comparatively evaluated, and the best degradation rate of IPR was presented by Achromobacter sp. C1 with a half-life (T1/2) of 9 days. Based on a nano-LC-MS/MS analysis for the strains, proteins solely expressed in the IPR treatment were identified by highlighting the strain Achromobacter sp. C1, with 445 proteins primarily involved in the biosynthesis of secondary metabolites and microbial metabolism in diverse environments. Differentially expressed protein amidases were involved in six metabolic pathways. Interestingly, formamidase was inhibited while other cyclases, i.e., amidase and mandelamide hydrolase, were overexpressed, thereby minimizing the effect of IPR on the metabolism of strain C1. The dynamic changes in the protein profiles of bacteria that degrade IPR have been poorly studied; therefore, our results offer new insight into the metabolism of IPR-degrading microorganisms, with special attention paid to amidases.

Effect of two broad-spectrum fungicides on the microbial communities of a soil subjected to different degrees of water erosion

The impact of both fungicide and water erosion on soil microbial communities has been little studied, and far less in subhumid-dry climates. To help fill this knowledge gap, we evaluated the influence of two broad-spectrum fungicides applied to soil subjected to different degrees of water erosion on microbial population and function. Soil samples were taken from an experimental site under three water erosion levels: an agricultural land subjected to minimum water erosion by terrace farming (MIN), an agricultural land subjected to moderate water erosion without terrace farming (MOD), and a land with no water erosion control treatment (non-eroded, NON). Three dosages of both carbendazim and iprodione were applied to microcosms, and the responses of microbial community and function were examined after 30-day exposure. Phospholipid fatty acid (PLFA) analysis, soil enzyme activities, and microbial abundances estimated by quantitative real-time PCR (qPCR) were used to evaluate the impact of carbendazim and iprodione fungicides applied at three dosages on microbial community structure and enzyme activities. The lowest values of fluorescein diacetate hydrolase (FDA), dehydrogenase, urease, and glucosidase activities were registered in soil under moderate erosion, while the highest ones were observed in non-eroded soils. Both carbendazim and iprodione decreased C- and P-related activities, even when those fungicides were applied at field rate doses. Thus, phosphomonoesterase, glucosidase, and xylosidase in soil treated with fungicides applied at recommended field rate decreased by 46 %, 61 %, and 34 % compared to soil without fungicide application. The lowest values of PLFA biomarkers for Gram-positive bacteria, Gram-negative bacteria, and arbuscular mycorrhizal fungi were registered in soils treated with the highest dose of both carbendazim and iprodione. Thus, Gram-positive bacteria, Gram-negative bacteria, arbuscular mycorrhizal fungi, and fungal PLFA biomarkers in soils without fungicide application were about 36 %, 3 %, 47 %, and 82 % higher than those soils treated with the highest doses of fungicides, respectively. In addition, the highest doses of fungicide application tended to greater decrease total PLFAs in both non-eroded and minimum eroded compared to moderate eroded soils. Similarly, the response of bacterial and fungal populations to fungicide treatment was more pronounced in non-eroded soils than in eroded soils. This study illustrates how the interactions between fungicides and water erosion levels affect soil enzyme activities, microbial abundances, and microbial community structure in agroecosystems.

Inhibition Analysis and High-Resolution Crystal Structure of Mus musculus Glutathione Transferase P1-1.

Multidrug resistance is a significant barrier that makes anticancer therapies less effective. Glutathione transferases (GSTs) are involved in multidrug resistance mechanisms and play a significant part in the metabolism of alkylating anticancer drugs. The purpose of this study was to screen and select a lead compound with high inhibitory potency against the isoenzyme GSTP1-1 from Mus musculus (MmGSTP1-1). The lead compound was selected following the screening of a library of currently approved and registered pesticides that belong to different chemical classes. The results showed that the fungicide iprodione [3-(3,5-dichlorophenyl)-2,4-dioxo-N-propan-2-ylimidazolidine-1-carboxamide] exhibited the highest inhibition potency (ΙC50 = 11.3 ± 0.5 μΜ) towards MmGSTP1-1. Kinetics analysis revealed that iprodione functions as a mixed-type inhibitor towards glutathione (GSH) and non-competitive inhibitor towards 1-chloro-2,4-dinitrobenzene (CDNB). X-ray crystallography was used to determine the crystal structure of MmGSTP1-1 at 1.28 Å resolution as a complex with S-(p-nitrobenzyl)glutathione (Nb-GSH). The crystal structure was used to map the ligand-binding site of MmGSTP1-1 and to provide structural data of the interaction of the enzyme with iprodione using molecular docking. The results of this study shed light on the inhibition mechanism of MmGSTP1-1 and provide a new compound as a potential lead structure for future drug/inhibitor development.

Investigation of the antifungal activity of the dicarboximide fungicide iprodione against Bipolaris maydis

Southern corn leaf blight (SCLB), mainly caused by Bipolaris maydis, is a destructive disease of maize worldwide. Iprodione is a widely used dicarboximide fungicide (DCF); however, its antifungal activity against B. maydis has not been well studied until now. In this study, the sensitivity of 103 B. maydis isolates to iprodione was determined, followed by biochemistry and physiology assays to ascertain the fungicide's effect on the morphology and other biological properties of B. maydis. The results indicated that iprodione exhibited strong inhibitory activity against B. maydis, and the EC50 values in inhibiting mycelial growth ranged from 0.088 to 1.712 μg/mL, with a mean value of 0.685 ± 0.687 μg/mL. After treatment with iprodione, conidial production of B. maydis was decreased significantly, and the mycelia branches increased with obvious shrinkage, distortion and fracture. Moreover, the expression levels of the osmotic pressure-related regulation genes histidine kinase (hk) and Ssk2-type mitogen-activated protein kinase (ssk2) were upregulated, the glycerin content of mycelia increased significantly, the relative conductivity of mycelia increased, and the cell wall membrane integrity was destroyed. The in vivo assay showed that iprodione at 200 μg/mL provided 79.16% protective efficacy and 90.92% curative efficacy, suggesting that the curative effect was better than the protective effect. All these results proved that iprodione exhibited strong inhibitory activity against B. maydis and provided excellent efficacy in controlling SCLB, indicating that iprodione could be an alternative candidate for the control of SCLB in China.

Genome-wide identification of the Penicillium digitatum bZIP gene family and the roles of one key member, PdatfA

Penicillium digitatum is the most common cause of postharvest decay in citrus fruits around the world. Previous studies revealed that the bZIP gene family plays crucial roles in development, stress adaptation, and pathogenicity in fungi. However, little is known about the bZIP genes in P. digitatum. In this study, we systematically identified the bZIP family in 23 Penicillium species and analyzed their evolutionary relationships. We found that gene loss and gene duplication shaped the evolution of the Penicillium bZIP family. P. digitatum experienced 3 bZIP gene loss events, but with no gene duplication. We subsequently characterized the biological functions of one important member, PdatfA in P. digitatum by constructing the deletion mutant. Results showed that ΔPdatfA exhibited a moderate growth defect, reduced pigmentation, and slightly increased resistance to fungicides iprodione and fludioxonil. However, ΔPdatfA displayed similar rot symptoms to that of the wild-type. The ΔPdatfA mycelia were not affected in response to oxidative stress while its conidia showed enhanced resistance due to the upregulation of catalases. Our results provide new insights into the evolution and functions of the bZIP gene family in Penicillium.

Impact of foliar application of fungicides on tomato leaf fungal community structure revealed by metagenomic analysis.

Fungicides are commonly used to manage plant pathogens. However, little is known about their effects on the non-target fungal communities that inhabit inside and outside the plant. These fungicides may have adverse effects on beneficial microbial communities with possible consequences for plant health and productivity. Hence, a metagenomic approach, based on the ITS2 region of fungal rDNA, was used to study the impact of foliar application of two fungicides (propineb and iprodione + carbendazim) on non-target tomato leaf fungal communities, in the context of early blight disease management. Metagenomic analysis revealed that the richness and diversity of tomato leaf fungal populations were adversely affected by the chemical treatments tested. Among the two fungicides, propineb (contact fungicide) imparted less non-targeted microorganisms than iprodione + carbendazim (systemic fungicide). In addition, all samples showed association of pathogenic genera Cladosporium, Corynespora, Pseudocercospora along with early blight pathogen Alternaria on tomato leaves that otherwise were undetected. Metagenomic studies also revealed a new mode of action for fungicides and bioagents besides their direct effect that is shifting the microbial community structure so that it provides greater resistance against the pathogen.

Iprodione and chlorpyrifos induce testicular damage, oxidative stress, apoptosis and suppression of steroidogenic- and spermatogenic-related genes in immature male albino rats.

The fungicide iprodione (IPR) and the insecticide chlorpyrifos (CPF) are concurrently applied for early disease control in fruits and other crops. However, there are no available data about the impacts of their co-exposure. Additionally, IPR and CPF are known as endocrine disruptors that can cause reproductive toxicity. The outcomes of their co-exposure on the development of male reproductive organs are still unknown. Therefore, this study aimed to assess the risk of exposure to these pesticides, particularly on the postnatal development of the male albino rat reproductive system from postnatal days 23-60. The results revealed that a single IPR or CPF exposure has harmful consequences on the reproductive development and function manifested by reduced testicular weight, serious changes in sperm characteristics, reproductive hormone level imbalance, testicular enzymes, oxidative stress and apoptosis-related enzymes, which correlated with transcription levels of steroidogenic- and spermatogenic-related genes. Histopathologically, both compounds caused severe damage in the testis and accessory glands architecture. Notably, co-exposure to IPR and CPF in rats caused more serious damage, indicative of an additive effect than individual exposure, so concurrent exposure should be avoided as it is more hazardous, especially on male fertility.

Dose-independent genotoxic response in A549 cell line exposed to fungicide Iprodione.

The fungicide Iprodione is widely applied in vegetables and raises concern for human health. The A549 human lung carcinoma cell line is a suitable model for assessing the toxicological effects of drugs. The goal of this work was to evaluate the genotoxicity and oxidative stress in the A549 cell line exposed to sublethal concentrations from 3 to 100µg/mL Iprodione considering LC50 = 243.4µg/mL Iprodione, as determined by the MTT assay. Generalized Linear Mixed Models (GLMM) were performed to determine the association between the responses NDI, MNim and MNib and the explanatory variables. Iprodione and solvent were relativized to the control whereas the concentration was included as numeric variable. ANOVA was used for the comparison of treatments. The coefficients of linear association between the explanatory variables and NDI, and the coefficients of logistic association between explanatory variables and MNim were not significant. However, these coefficients showed significant association with MNib only for Iprodione treatment but not for Iprodione concentration, indicating lack of dose-response relationship. Genotoxicity risk assessment indicated that the increase in Iprodione concentrations increased slightly the probability of belonging to the genotoxic category. ANOVA showed significant differences in MNib, and non-significant differences in NDI and MNim among treatments. The oxidative stress analysis performed at 3, 12, and 25μg/mL Iprodione showed a significant and linear increase in SOD, and a significant and linear decrease in GSH and GST. The Dunnett test was significant for GSH at 12 and SOD at 25μg/mL.

N-acetylcysteine reduced the immunotoxicity effects induced in vitro by azoxystrobin and iprodione fungicides in mice.

Azoxystrobin (AZO) and Iprodione (IPR) fungicides are extensively used worldwide, and therefore, contaminate all environmental compartments. The toxicity and the mechanisms by which they affected immune cells are complex and remain unknown. This study investigated the impact of AZO and IPR on the in vitro function of mice peritoneal macrophages including lysosomal enzyme activity and tumor necrosis factor (TNF)α and nitric oxide (NO) production in response to lipopolysaccharide (LPS) stimulation, the proliferation of mice splenocytes stimulated by concanavalin (Con)A and LPS, and the production of the Th1cytokine interferon-gamma (IFNγ) and the Th2 cytokine interleukin (IL)-4 and IL-10 by ConA-activated splenocytes. This is the first report indicating that AZO and IPR fungicides dose-dependently inhibited mice macrophage lysosomal enzyme activity and LPS-stimulated production of TNFα and NO. Mitogen-induced proliferation of mice splenocytes was also suppressed by AZO and IPR in a dose-dependent manner. More pronounced impact was observed on ConA-induced response. The production of IFNγ by ConA-stimulated splenocytes was dose-dependently inhibited; however, the production of IL-4 and IL-10 increased in the same conditions. These results suggested that AZO and IPR polarized Th1/Th2 cytokine balance towards Th2 response. Overall, marked immunosuppressive effects were observed for AZO. The immunomodulatory effects caused by AZO and IPR were partially reversed by the pharmacological antioxidant N-acetylcysteine (NAC), suggesting that both fungicides exerted their actions through, at least in part, oxidative stress-dependent mechanism. Collectively, our data showed that AZO and IPR fungicides exerted potent immunomodulatory effects in vitro with eventually strong consequences on immune response and immunologically based diseases.

Postharvest quality and brown rot incidence in plums treated with Ascophyllum nodosum extract

The use of extracts of algae in agriculture has been growing because of their benefits to plant development and production. The objective of this study was to evaluate the effects of Ascophyllum nodosum extract on the postharvest quality of ‘Irati’ and ‘Reubennel’ plums and on the incidence of brown rot. The treatments consisted of concentrations of 0, 0.1, 0.2, 0.3 and 0.4 mL L-1 of a commercial product containing algae extract (Algamare®) that were applied to the fruits by immersion. The fruits were then stored for 15 (cv. Irati) or 30 days (cv. Reubennel) at 2.5 °C and 90% RH and for two additional days at ambient conditions for further physicochemical analysis. A batch of ‘Reubennel’ plums was inoculated with Monilinia fructicola and kept for seven days in a cold room and an additional nine days at room temperature to evaluate the occurrence of brown rot. The postharvest application of the extract at a concentration of 0.4 mL L-1 altered some of the physicochemical characteristics of the fruit after storage; an increase in the total phenolic compound content and PG activity were observed in ‘Irati’ plums. Reubennel fruits had higher titratable acidity, a lower TSS/TA ratio, lower pulp firmness and higher PME activity. The most significant effect was observed with the application of A. nodosum extract at 0.4 mL L-1, which significantly reduced the incidence and severity of brown rot in the inoculated fruits and had a similar control effect as the application of the fungicide iprodione.

The response of soil and phyllosphere microbial communities to repeated application of the fungicide iprodione: accelerated biodegradation or toxicity?

Pesticides interact with microorganisms in various ways with the outcome being negative or positive for the soil microbiota. Pesticides' effects on soil microorganisms have been studied extensively in soil but not in other pesticides-exposed microbial habitats like the phyllosphere. We tested the hypothesis that soil and phyllosphere support distinct microbial communities, but exhibit a similar response (accelerated biodegradation or toxicity) to repeated exposure to the fungicide iprodione. Pepper plants received four repeated foliage or soil applications of iprodione, which accelerated its degradation in soil (DT50_1st = 1.23 and DT50_4th = 0.48 days) and on plant leaves (DT50_1st >365 and DT50_4th = 5.95 days). The composition of the epiphytic and soil bacterial and fungal communities, determined by amplicon sequencing, was significantly altered by iprodione. The archaeal epiphytic and soil communities responded differently; the former showed no response to iprodione. Three iprodione-degrading Paenarthrobacter strains were isolated from soil and phyllosphere. They hydrolyzed iprodione to 3,5-dichloraniline via the formation of 3,5-dichlorophenyl-carboxiamide and 3,5-dichlorophenylurea-acetate, a pathway shared by other soil-derived arthrobacters implying a phylogenetic specialization in iprodione biotransformation. Our results suggest that iprodione-repeated application could affect soil and epiphytic microbial communities with implications for the homeostasis of the plant-soil system and agricultural production.

Comprehensive assessment for the residual characteristics and degradation kinetics of pesticides in Panax notoginseng and planting soil

Incorrect and excess usage of pesticides during crop cultivation poses a serious threat to human health and ecosystems. In this study, we tested for the presence of 201 pesticide residues in 90 batches of Panax notoginseng (P. notoginseng) and 10 batches of planting soil. Pesticide residue characteristics and the relationship between pesticides present in P. notoginseng and the soil were discussed. Twenty-nine pesticides were detected in P. notoginseng samples and 15 pesticides were found in the soil samples. In P. notoginseng samples, the 68.9% of the identified pesticides were fungicides, and six fungicides (procymidone, iprodione, pyrimethanil, propiconazole, dimethomorph and tebuconazole) were found in >90% of the samples. Nine insecticides were found, with one insecticide, chlorpyrifos, detected in 93.3% of the P. notoginseng samples. The residual concentrations of 17 pesticides were found at levels exceeded the “non-Chinese” maximum residue levels (MRLs) for Ginseng and 17 pesticides were found at levels exceeding the MRLs set by China for “pollution-free” P. notoginseng. We observed no significant differences in pesticide residues were found on P. notoginseng from different cultivation areas. We also analyzed the degradation kinetics of pesticides in the soil, as well as their bioconcentration factors (BCFs), and found that the fungicides iprodione and myclobutanil displayed strong uptake from the soil to the root of P. notoginseng. Together, our data suggest that fungicides should be considered as key monitoring substances in P. notoginseng and planting soil.

Potential of Certain Salts and Fungicides to Control Postharvest Gray Mould (B. cinerea) of Strawberry

Effect of two fungicides (i.e. iprodione and cyprodinil+flydioxonil mixture) and two salts (sodium bicarbonate, SBC, and sodium benzoate, SBO) were evaluated separately or in mixtures in laboratory experiments as well as under field conditions against Botrytis cinerea instant of the fruit gray mould of strawberry. All tested salts and fungicides as well as their mixtures significantly decreased the radial growth of B. cinerea in vitro. Complete inhibition of radial growth was obtained with SBC at 2.0 and1.0 g/100ml. Also, high reductions were obtained with SBC at lower concentrations. The fungicide iprodione at 500 mg/L completely inhibited radial growth of B. cinerea, while complete inhibition was obtained with cyprodinil+flydioxonil at lower concentration (10 mg/L). Mixing the tested fungicides with salts increased their inhibition effect on B. cinerea growth at lower fungicide concentrations. iprodione at 0.1 mg/L mixed with SBO at 1g/100ml completely inhibited radial growth. Also, cyprodinil+flydioxonil at 0.1mg/L mixed with SBC at 0.2 g/100ml completely inhibited radial growth of B. cinerea while 0.001mg/L cyprodinil+flydioxonil fungicide with SBO at 2.0g/100 completely inhibited radial growth. On the other hand, the field experiments showed that all treatments with the tested salts and fungicides significantly decreased postharvest Botrytis gray mould on fruits harvested 1, 3, 7, 10 and 15 days after treatments and stored at 5˚C for seven days. However, the treatment with 50% cyprodinil+flydioxonil + SBC 2% was the most effective and decreased mean percentage of infection to 4% compared to 23.86 % for the untreated control. This was followed by 50% iprodione +SBC 2%, 25% iprodione + SBC and 25% cyprodinil+flydioxonil + SBC 2%, and cyprodinil+flydioxonil, alone, and iprodione, alone, with percentages of infection being 7%, 8%, 8.5%, 8.5%, and 9%, respectively, while (SBC 2%, alone) treatment showed the least effect with 12.93 percentage of infection. Meanwhile, positive strong correlations (r = 0.981 to 0.943) were revealed between Infection (%) and harvest interval periods with two salts mixed with iprodione and cyprodinil + flydioxonil after stored at 5˚C. Also, the enzyme assay revealed that increases in the activity levels of pectolytic and cellulytic enzymes produced by the fungus were associated with high fruit rots incited by B. cinerea. Meanwhile, residues of all tested fungicides in treated strawberry fruits decreased with decreasing fungicide concentrations in the salt + fungicide mixtures. cyprodinil (25%) + SBO, and cyprodinil (25%) + SBC treatments exhibited the lowest residue values being 1.420, and 1.680 mg/kg fruit one hour after treatment while the other treatments showed higher residue values while the highest was recorded for iprodion at recommend dose being 29.81 mg/kg fruit. However, fungicide residues decreased with increasing time after preharvest treatments.

The fungicide iprodione affects midgut cells of non-target honey bee Apis mellifera workers.

The honey bee Apis mellifera is an important pollinator of agricultural crops and natural forests. Honey bee populations have declined over the years, as a result of diseases, pesticides, and management problems. Fungicides are the main pesticides found in pollen grains, which are the major source of protein for bees. The objective of this study was to evaluate the cytotoxic effects of the fungicide iprodione on midgut cells of adult A. mellifera workers. Bees were fed on iprodione (LD50, determined by the manufacturer) for 12 or 24h, and the midgut was examined using light and transmission electron microscopies. The expression level of the autophagy gene atg1 was assessed in midgut digestive cells. Cells of treated bees had signs of apoptosis: cytoplasmic vacuolization, apical cell protrusions, nuclear fragmentation, and chromatin condensation. Ultrastructural analysis revealed some cells undergoing autophagy and necrosis. Expression of atg1 was similar between treated and control bees, which can be explained by the facts that digestive cells had autolysosomes, whereas ATG-1 is found in the initial phases of autophagy. Iprodione acts by inhibiting the synthesis of glutathione, leading to the generation of reactive oxygen species, which in turn can induce different types of cell death. The results indicate that iprodione must be used with caution because it has side effects on non-target organisms, such as pollinator bees.

Molecularly imprinted sol-gel polymers for the analysis of iprodione fungicide in wine: Synthesis in green solvent

Iprodione is a fungicide widely used in viticulture in most agricultural countries. It was banned recently in the European community because of its carcinogenic and endocrine disrupting characters. In this work, a cheap analytical method able to monitor iprodione in a white wine was developed. Molecularly imprinted sol-gel polymers (MIS) specific to iprodione and using green solvents were synthesized. An experimental design having the following factors (solvent volume and crosslinker quantity) was used to prepare an optimal MIS. In terms of selectivity, the optimal MIS showed the best partition coefficient towards iprodione in a white wine containing four other competing fungicides (procymidone, pyrimethanil, azoxystrobin and iprovalicarb). A solid phase extraction method using the optimal MIS was optimized and applied to analyse iprodione in a white wine. Low detection and quantification limits were reached 11.7 and 39.1 µg/L respectively.

PGPBS, a mitogen-activated protein kinase kinase, is required for vegetative differentiation, cell wall integrity, and pathogenicity of the barley leaf stripe fungus Pyrenophora graminea

The high-osmolarity glycerol (HOG) signaling pathway regulates the adaptation of fungi to environmental stressors. The mitogen-activated protein kinase kinase (MAPKK) PBS2 of Saccharomyces cerevisiae serves as a scaffold protein in the HOG pathway. We characterized the pgpbs gene of Pyrenophora graminea, which encodes a MAPKK that is 56% orthologous to PBS2 of S. cerevisiae. A cloning technique based on homology was applied to amplify the pgpbs gene. Specific silent mutations then were generated in pgpbs. We evaluated the potential roles of PGPBS in the osmotic response, vegetative differentiation, cell wall integrity, drug resistance, and pathogenicity. Our findings indicated that the pgpbs coding region comprises 2075 base pairs and encodes a protein of 676 amino acids. Mutants deficient in pgpbs expression had significant reductions in vegetative growth and were sensitive to calcofluor white (CFW), an inhibitor of cell wall synthesis. Mutants also lost pathogenicity and were sensitive to an osmotic stress–inducing medium containing NaCl and sorbitol. Moreover, mutants had increased resistance to the dicarboximide fungicide iprodione and the triazole fungicide tebuconazole. These findings suggest that pgpbs is involved in the osmotic and ionic stress responses, vegetative differentiation, cell wall integrity, virulence, and tolerance to iprodione and tebuconazole. We expect that our findings will help elucidate the pathogenesis of barley leaf stripe and will inform strategies for breeding resistance to this disease.

Resistance risk assessment for fludioxonil in Sclerotinia homoeocarpa in China

Sclerotinia homoeocarpa causes dollar spot disease on turfgrass and is a serious problem on many species worldwide. Fludioxonil, a phenylpyrrole fungicide, is not currently registered for dollar spot control in China. In this study, the baseline sensitivity to fludioxonil was established using an in vitro assay for 105 isolates of S. homoeocarpa collected from 10 locations in different regions of China. Results indicate that the frequency distribution of effective concentration for 50% inhibition of mycelial growth (EC50) values of the S. homoeocarpa isolates was unimodal (W = 0.9847, P = .2730). The mean EC50 value was 0.0020 ± 0.0006 μg/ml with a range from 0.0003 to 0.0035 μg/ml. A total of 7 fludioxonil-resistant mutants were obtained in laboratory, the mutants were stable in fludioxonil sensitivity after the 10th transfer, with resistance factor (RF) ranging from 4.320 to >13,901.4. The mutants showed a positive cross-resistance between fludioxonil and the dicarboximide fungicide iprodione, but not propiconazole, fluazinam, and thiophanate-methyl. When mycelial growth rate, pathogenicity and osmotic sensitivity were assessed, the mutants decreased in the fitness compared with their parental isolates. Sequence alignment of the histidine kinase gene Shos1 revealed a 13-bp fragment deletion only in one mutant, no mutations were observed on Shos1 in the rest resistant mutants.