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Botryosphaeria dothidea causes white rot, which is among the most devastating diseases affecting apple crops globally. In this study, we assessed B. dothidea resistance to carbendazim by collecting samples from warts on the infected branches of apple trees or from fruits exhibiting evidence of white rot. All samples were collected from different orchards in nine provinces of China in 2018 and 2019. In total, 440 B. dothidea isolates were evaluated, of which 19 isolates from three provinces were found to exhibit carbendazim resistance. We additionally explored the fitness and resistance stability of these isolates, revealing that they were no less fit than carbendazim-sensitive isolates in terms of pathogenicity, sporulation, and mycelial growth and that the observed carbendazim resistance was stable. Sequencing of the β-tubulin gene in carbendazim-resistant isolates showed the presence of a substitution at codon 198 (GAG to GCG) that results in an alanine substitution in place of glutamic acid (E198A) in all 19 resistant isolates. A loop-mediated isothermal amplification (LAMP) method was then developed to rapidly and specifically identify this E198A mutation. This LAMP method offers value as a tool for rapidly detecting carbendazim-resistant isolates bearing this E198A mutation and can thus be used for the widespread monitoring of apple crops to detect and control the development of such resistance.

Sclerotinia sclerotiorum is one of the most damaging and economically important necrotrophic plant pathogens, infecting more than 400 plant species globally. Although the phenylpyrrole fungicide fludioxonil has high activity against S. sclerotiorum, reports indicate that there is also substantial potential for the development of fungicide resistance. However, the current study investigating five fludioxonil-resistant laboratory mutants found a significant fitness cost associated with fludioxonil resistance resulting in significantly (P < 0.05) reduced mycelial growth and sclerotia formation on potato dextrose agar as well as significantly (P < 0.05) lower pathogenicity on detached tomato leaves, with one mutant, LK-1R, completely losing the capacity to cause infection. In addition, all of the fludioxonil-resistant mutants had significantly (P < 0.05) increased sensitivity to osmotic stress (0.5 M of potassium chloride and 1.0 M of glucose), which is consistent with the proposed fludioxonil target sites within the high osmolarity glycerol stress response mitogen-activated protein kinase (HOG1-MAPK) signaling transduction pathway. Sequence analysis of six genes from this two-component pathway, including SsHk, SsYpd, SsSk1, SsSk2, SsPbs, and SsHog, revealed several mutations that may be associated with fludioxonil resistance. For example, six separate point mutations were found in SsHk that led to changes in the predicted amino acid sequence, including A136G, F249V, G353A, E560K, M610K, and K727R. Similarly, SsPbs had three mutations (D34G, S46L, and L337E), SsSk1 and SsYpd had two (S53G and A795V for SsSk1, and E67G and Y141H for SsYpd), and SsHog and SsSk2 had one each (V220A and S763P, respectively). To our knowledge, these constitute the first reports of amino acid changes in proteins of the HOG1-MAPK pathway being associated with fludioxonil resistance in S. sclerotiorum. This study also showed a positive cross-resistance between fludioxonil and dimethachlone and procymidone, but none with tebuconazole or carbendazim, indicating that the inclusion of tebuconazole within an integrated pest management program could reduce the risk of fludioxonil resistance developing in field populations of S. sclerotiorum and ensure the sustainable production of soybeans in China into the future.

Background Ceratocystis fimbriata is a hazardous fungal pathogen able to cause black rot disease on sweet potato. The management of C. fimbriata strongly relies on the use of toxic fungicides, and there is a lack of efficient alternative strategies. Results The antifungal properties of quinolinic acid (QA) were studied for the first time, indicating that QA shows selective antifungal activity against C. fimbriata. QA inhibited completely the mycelial growth of C. fimbriata at less than 0.8 mg mL-1 concentration (pH 4), and was able to produce alterations in the fungal cell wall, and to impede spore agglutination and mycelium formation. QA significantly reduced the concentration of ergosterol, and was able to associate to iron (II), suggesting that QA may be a lanosterol 14-α demethylase inhibitor. In preventive applications, QA reduced the disease incidence of C. fimbriata on sweet potato by 75%, achieving higher control efficacy in comparison with commercial fungicides prochloraz and carbendazim. Conclusions The first selective antifungal agent against C. fimbriata was discovered in this work, and showed suitable antifungal properties for the management of black rot disease. © 2021 Society of Chemical Industry.

The occurrence of pesticide residues in bananas imported from Ecuador and the Philippines into Saudi Arabia was investigated. Eighty-seven banana samples were purchased from various supermarkets around the country. The conventional QuEChERS method was used to extract 294 pesticides in whole bananas and the analysis was carried out using LC-MS/MS and GC-MS/MS. In total, nine pesticides were detected of which azoxystrobin, carbendazim, chlorpyrifos, imazalil, and thiabendazole were the most frequently found in bananas from both countries. The levels of these pesticides were complaint according to the Saudi Food & Drug Authority’s (SFDA) maximum residue levels (MRLs), therefore they cannot be considered a public health problem. Detectable residues of these agrochemicals show that they are used extensively in banana cultivation. The Ecuadorian Agency for Agricultural Quality Assurance (Agrocalidad) and the Saudi Ministry of Environment, Water and Agriculture (MEWA) prohibit the use of carbendazim in agriculture, while the Filipino Fertiliser and Pesticide Authority (FPA) allows its use. A legislative comparison of pesticide approval status revealed a lack of consensus between banana exporting and importing countries that could affect food safety and trade.

To improve the effect of pine rosin in plant fungicides, four series of dehydroabietyl-1,3,4-thiadiazole derivatives from the natural product rosin were synthesized. Based on the evaluation of the in vitro antifungal activity against Sclerotinia sclerotiorum, Botrytis cinerea, Fusarium oxysporum, and Magnaporthe oryzae, rosin-based 1,3,4-thiadiazole compounds containing thiophene heterocycles were screened. Notably, compound 3e [dehydroabietyl-(1,3,4-thiadiazol-2-yl)-5-nitrothiophene-2-carboxamide] exhibited excellent antifungal property against F. oxysporum with an EC50 of 0.618 mg/L, which was lower than that of the positive control carbendazim (0.649 mg/L). The in vivo antifungal activity results showed that 3e exerted a protective effect on cucumber plants. Physiological and biochemical studies showed that the primary mechanism of action of compound 3e on F. oxysporum was it changed the mycelial morphology, increased the cell membrane permeability, and inhibited the synthesis of ergosterol in the mycelia. Furthermore, the quantitative structure-activity relationship studies revealed that the frontier orbital energy in the molecule had a key role in the antifungal activity through the conjugation and electrostatic interaction between compound 3e and the receptors of the target. Thus, the present study highlighted the application of rosin-based fungicidal candidates and exploited efficient plant pesticides for sustainable crop production.

This study aims to encapsulate the fungicide carbendazim using a biodegradable polymer (pectin). First, we have obtained calcium pectinate beads (CPG-Carb) by ionotropic gelation using calcium ions as a crosslinking agent. These beads were then coated with silica starting from tetraethoxysilane (TEOS), by a sol-gel process to form hybrid beads (CPG-Carb-SG). The morphology, composition and structure of both beads were characterized and the controlled release assays of the fungicide were studied in both water and soil columns. The encapsulation efficiency for CPG-Carb was slightly higher (75%) compared to CPG-Carb-SG (67%) due to carbendazim loss during the impregnation and condensation steps. The release rate in water and soil columns was about 4 times lower for CPG-Carb-SG than CPG-Carb demonstrating the efficiency of the silica coating to delay the release of carbendazim. Moreover, the release of CPG-Carb-SG is due to the erosion of the silica layer during the first two weeks. After this period, the silica layer was degraded, and the release is then controlled by the swelling of the organic part of the bead as observed for CPG-Carb. Finally, the biodegradability of the pectin, and the release profile make such systems promising candidates for sustained and economical pesticide delivery systems.

Recently, cost-effective and rapid analysis of carbendazim, in surface, drinking and ground water has attracted a huge deal of interest. Nevertheless, for given applications, the use of present biological receptor-based sensing approaches is limited based on the costs and stability. Hence, the best alternatives are the cost-effective and robust imprinted polymeric receptors. The current study deals with synthesizing MIPs (molecularly imprinted polymers) for carbendazim utilizing methacrylic acid in chloroform as a porogen. The MIP’s binding affinity with carbendazim was assessed in aqueous media. Superior imprinting features for carbendazim were found by MIPs synthesized in chloroform. Imprinted polymers were prepared using bulk polymerization, by mixing MAA as a functional monomer, carbendazim as a template, EGDMA as a cross-linker and AIBN as a initiator were dissolved in chloroform. Here, it was demonstrated carbendazim MIPs merged into a CP (carbon paste) electrode as a MIP-CP sensor platforms to detect the carbendazim. By this simple sensor format, precise determining of carbendazim is allowed utilizing an electrochemical method. We have achieved a detection limit of 3.1 × 10−8 mol L−1 for this compound in linear range of 0.1–100 μM. By the simple procedure, low production cost, and easy preparation, the MIP sensor becomes attractive for sensitive and selective detection of analytes, even in less-armed laboratories with nominal training.

Wilt caused by Fusarium oxysporum f. sp. melongenae causes high yield loss in eggplant (Solanum melongena L.). The present research was conducted to evaluate the effectiveness of combined use of Tr...

Graphical abstract, An efficient protocol for in-vitro propagation of an important ornamental crop, Petunia hybrida Vilm. Cv. “Bravo” was developed. The explants that were used to carry out the experiment were Leaf segments, nodal segments and shoot tips. Nodal segments recorded highest per cent asepsis followed by shoot tips and leaf segments. Asepsis was found to be highest when the explants were sterilized with Fungicide (Carbendazim) 0.02% for the duration of 30 min followed by 0.1% HgCl2 for duration of 10 min and then ethanol 70% for 10 s. Longer duration of the sterilant treatment showed more necrotic effects on the explants, thus mercuric chloride treatment when given for 5 min proved to be more effective in terms of survival of the explants. Maximum establishment per cent was recorded in Murashige and Skoog (MS) media fortified with BAP (1.5 mg L−1) and IBA (0.5 mg L−1) in shoot tips and nodal segments, i.e. 97.90 and 95.74% respectively. Callus was efficiently induced and developed when PGR amalgamation of BAP (0.1 mg L−1) and 2,4-D (1.5mg L−1) was used. Kinetin at the concentration of 2.0 mg L−1 along with IBA at 0.5mg L−1 recorded highest callus regeneration in both leaf and internodal segment derived callus. Maximum proliferation percent of shoots (97.90%), highest number of shoots (20.50 explant−1) and maximum length of shoot (2.70 cm) was recorded in PGR combination of IBA and BAP both at 0.5 mg L−1 concentration level. Rhizogenesis was recorded to be highest in the MS media containing IBA 1.00 mg L−1. Best hardening media which recorded maximum survival per cent 92.50% was noticed on the media formulation comprised of equal ratio of perlite and vermiculite mix, under poly house conditions.

Biodiversity is under pressure worldwide, with amphibians being particularly threatened. Stressors related to human activity, such as chemicals, are contributing to this decline. It remains, however, unclear whether chemicals exhibiting a fungicidal activity could indirectly affect tadpoles that depend on microbially conditioned leaf litter as food source. The indirect effect of fungicides (sum concentration of a fungicide mixture composed of azoxystrobin, carbendazim, cyprodinil, quinoxyfen, and tebuconazole: 100 µg/L) on tadpoles was assessed relative to leaf litter colonized by microbes in absence of fungicides (control) and a worst‐case scenario, that is leached leaf litter without microbial colonization. The quality of leaf litter as food for tadpoles of the European common frog (Rana temporaria) was characterized through neutral lipid fatty acid profiles and microbial sum parameters and verified by sublethal responses in tadpoles (i.e., feeding rate, feces production, growth, and fatty acid composition). Fungicides changed the nutritious quality of leaf litter likely through alterations in leaves’ neutral lipid fatty acid profiles (i.e., changes in some physiologically important highly unsaturated fatty acids reached more than 200%) in combination with a potential adsorption onto leaves during conditioning. These changes were reflected by differences in the development of tadpoles ultimately resulting in an earlier start of metamorphosis. Our data provide a first indication that fungicides potentially affect tadpole development indirectly through bottom‐up effects. This pathway is so far not addressed in fungicide environmental risk assessment and merits further attention., Tadpoles consuming leaf litter of different quality show changes in their energy reserves, more specifically the composition of fatty acids, and growth dynamics. Low‐quality food ultimately leads to a higher number of individuals entering metamorphosis.

In the field of agriculture fungicides are vital, providing the most important ecosystem service for food production. The widespread use of these chemicals can significantly lead to various ecotoxicological threats with adverse effects, such as environmental changes, microbial resistance, and phytotoxicity. Electrochemical sensors offer enormous potential for the identification and monitoring of hazardous substances in accordance with their constructive characteristics, namely, precision, accuracy, sensitivity, and selectivity, over traditional analytical techniques. Here, we thus report the synthesis of the lanthanum vanadate/hexagonal boron nitride (LaV/h-BN) composite for the electrochemical determination of carbendazim (CZ), which is a widely used fungicide for disease management with critical risks associated with its overexposure. The combination of LaV and h-BN accelerates the formation of active sites, facilitating faster charge transfer and higher electronic conductivity. The synergistic effects greatly improve the preference of the modified electrode with increased sensitivity, a lower limit of detection, and wide linear responses toward CZ detection. The existence of variable oxidation states in the orthovanadate together with the unique properties of h-BN mark LaV/h-BN as an advanced material for specific applications in the family of rare-earth metal orthovanadate. Also, the deep eutectic solvent-assisted synthesis of the material creates an environmentally efficient system with reduced energy requisites to allow for applications in effective environmental monitoring.

Gray mold, caused by the fungus Botrytis cinerea Pers ex Fr., is one of the most destructive spoilage diseases, severely affecting tomato production in Henan Province, China. Spraying fungicides from the flowering to the harvest stage is a necessary measure to reduce losses associated with B. cinerea infection. However, B. cinerea has developed resistance to fungicides in many countries. Boscalid is a succinate dehydrogenase inhibitor (SDHI) fungicide and was registered for the control of gray mold. In this study, a total of 269 B. cinerea isolates were collected from tomato in commercial greenhouses in different locations of Henan Province in 2014 and 2015. The sensitivity and resistance of B. cinerea field isolates were determined based on mycelial growth. The effective concentration 50 ranged from 0.11 to 15.92 µg/ml and 0.16 to 8.54 µg/ml, in 2014 and 2015, respectively. The frequency of low resistance to boscalid was 12.6 and 7.6%, and moderate resistance was 2.7 and 1.3% in 2014 and 2015, respectively. No highly resistant isolates were found in Henan Province, China. Mycelial growth, mycelial dry weight, spore production, and pathogenicity were not significantly different between resistant and sensitive phenotypes of the B. cinerea isolates. The results of cross-resistance testing showed no correlation between boscalid and carbendazim, procymidone, pyrimethanil, fluazinam, or fluopyram. In this study, the succinate dehydrogenase genes B (sdhB), C (sdhC), and D (sdhD) were analyzed and compared in sensitive and low-resistance and moderately resistant B. cinerea isolates to boscalid. Results showed that point mutations occurred simultaneously at sdhC amino acid positions 85 (G85A), 93 (I93V), 158 (M158V), and 168 (V168I) in 4 out of 10 sensitive isolates and 23 of 26 low-resistance and 5 of 5 moderately resistant B. cinerea isolates to boscalid. No point mutations were found in the sdhB and sdhD genes of all isolates. Furthermore, no point mutations were found in sdhB, sdhC, and sdhD genes in 3 of 26 low-resistance B. cinerea isolates to boscalid. Therefore, we speculate that the simultaneous point mutations in the sdhC gene may not be related to the resistance of B. cinerea to boscalid. These results suggested that there might be a substitution mechanism for the resistance of B. cinerea to the SDHI fungicide boscalid.

Bjerkandera adusta degrades polycyclic aromatic compound. However, its effect on preventing plant disease is unknown. We carried out both the confrontation culture and greenhouse pot experiments to address effects of B. adusta M1 strain on the growth of pathogenic Fusarium oxysporum f. sp. conglutinans (FOC) and incidence of Fusarium wilt in Brassica napus L. The results showed the hyphal growth rate of B. adusta M1 was significantly greater than that of FOC. The B. adusta M1 fermentation broth also significantly inhibited the growth of FOC hyphae by 62.79 ± 1.80%, which was greater than an inhibition rate of 40.63 ± 1.68% by the fungicide, carbendazim. The electron microscopic image showed the FOC hyphae was directly penetrated by the B. adusta M1 hyphae, indicating a strong mycoparasitism by B. adusta M1. Both the B. adusta M1 and its fermentation broth reduced the incidence and disease index of the Fusarium wilt in Br. napus, and the control effects of B. adusta M1 and its fermentation broth on Fusarium wilt were comparable or better than carbendazim. Furthermore, both the B. adusta M1 and its fermentation broth increased the activity of the defense enzymes including superoxide dismutase (25.92% and 22.38%), catalase (78.80% and 82.28%), peroxidase (46.64% and 6.83%) and phenylalanine ammonia-lyase (44.23% and 5.77%). Similarly, both the B. adusta M1 and its fermentation broth decreased the cell membrane permeability (48.68% and 33.71%) and malondialdehyde concentration (71.04% and 17.51%) of plant leaf. Therefore, B. adusta M1 can be applied as a potential biocontrol fungus to against Fusarium wilt.

The present study was focused on synthesis and characterization of copper nanoparticles to evaluate their efficacy against fruit rot pathogen of chilli crop. The green synthesis of nanoparticles was carried out by using extracts of Eucalyptus and Mint leaves. The synthesis of copper nanoparticles was confirmed by XRD, PSA, SEM and TEM. The average size of these particles synthesized by Eucalyptus leaf extract (CuNP-E) ranged from 10 to 130 nm, while as size of Mint leaf extract synthesized particles (CuNP-M) ranged from 23 to 39 nm, thus confirming their nano size. These green synthesized copper nanoparticles were evaluated against Colletotrichum capsici where Carbendazim 50 WP @ 500 ppm and copper oxychloride 50 WP @ 2500 ppm served as standard checks. The mycelia inhibition of Colletotrichum capsici caused by copper nanoparticles was studied on PDA medium. CuNP-M @ 1000 ppm showed highest mycelial inhibition of 99.78% followed by 93.75% at 500 ppm and CuNP-E @ 1000 ppm compared to standard fungicides, carbendazim 50 WP @ 500 ppm (72.82%), and copper oxychloride 50 WP @ 2500 ppm (85.85%). The CuNP-M @ 500 ppm were significantly superior to carbendazim 50 WP @ 500 ppm and copper oxychloride 50 WP @ 2500 ppm, but was statistically at par with CuNP-E @ 1000 ppm. This shows effectiveness of much lower concentration of copper nanoparticles compared to conventional fungicides. In detached fruit method, nanoparticles applied before inoculation of pathogen showed better results with regard to incubation period, lesion number and lesion size than after inoculation of pathogen. The present study reveals a simple, convenient, non-toxic and cost-efficient technique for the synthesis of nanoparticles and their effectiveness against Colletotrichum capsici. CuNP-M first time synthesized and evaluated against Colletotrichum capsici performed better than CuNP-E.

Fusarium verticillioides laboratory isolates resistant to carbendazim were readily obtained after mutagenesis with UV-irradiation. Three resistant phenotypes that included highly (HR, Resistant factors>30), moderately (MR, Rf: 14–25) and low (LR, Rf: 2–5) resistant isolates to carbendazim were identified by fungitoxicity tests in vitro. Mutant isolates were simultaneously resistant to thiabendazole and benomyl but not to the also tubulin-blocking fungicides diethofencarb and zoxamide. No cross resistance relationship was found between carbendazim and fungicides with different modes of action such as the dicarboximide iprodione, the QoI pyraclostrobin, the imidazole prochloraz, the triazoles flusilazole and epoxiconazole. Mutation(s) for resistance to benzimidazoles did not significantly affect mycelial growth rate whereas sporulation and pathogenicity were adversely affected in most of the resistant mutants. Moreover, low temperatures suppressed the expression of carbendazim-resistance in all resistant phenotypes. Most mutant isolates produced Fumonisin B1 in similar or less quantities than the wild-type isolate both in vitro and in vivo in the absence of carbendazim with the exception of one HR isolate which had up to 3-fold increased FB1 production. Sub lethal doses of carbendazim in the growth medium in vitro resulted in a significant reduction in FB1 levels in all cases even though most mutant isolates produced higher FB1 quantities than the wild type. A significant correlation was found between F. verticillioides sporulation and FB1 production on maize seeds. The potential risk of food and feed contamination by FB1 producing F. verticillioides isolates resistant to benzimidazoles and implications on resistance management strategies are discussed.

Macrophomina phaseolina (Tassi) Goid persists throughout the crop growth period and incurred substantial fiber yield losses in jute crop. This main aim of this research was to find out, potentiality of eight systemic and broad spectrum fungicide viz., (Azoxystrobin (strobilurin), Carbendazim (benzimidazole), Difenconazole, Tebuconazole, Propiconazole, Cyproconazole, Tricyclazole (triazole) and Propineb (propylene bis-dithiocarbamate), used either alone or with combination, as a seed treatment (ST) and foliar sprays (FS) to control M. phaseolina. Significant difference (p

In this study,the QuEChERSand high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS)was applied to determine thiophanate-methyl and its metabolite carbendazim residue in water...

Anthracnose, caused by Colletotrichum spp., is the most significant field and post-harvest disease of mango worldwide and is mainly controlled through the use of systemic fungicides belonging to the methyl benzimidazole carbamate (MBC) class. In Mexico, this disease in mango has been associated with at least seven cryptic species belonging to the C. gloeosporioides complex. In this study, the distribution of sensitivity to MBC (benomyl and carbendazim) and prochloraz fungicides of 101 Colletotrichum spp. isolates obtained from 101 commercial orchards in Mexico’s most important mango-producing states was determined. All Colletotrichum spp. isolates were obtained from commercial mango orchards with applications of MBC fungicides and that had not been exposed to applications of prochloraz, or any other DMI (demethylation inhibitors) class fungicide. Isolates were evaluated using an in vitro test to determine the effective fungicide concentration at which 50% of mycelial growth is inhibited (EC50) for each isolate-fungicide combination. EC50 values for benomyl and carbendazim ranged from 0.0967 to 0.3995 µg mL−1 (median = 0.1923 µg mL−1) and from 0.0880 to 0.1703 µg mL−1 (median = 0.1171 µg mL−1), respectively, while those for prochloraz ranged from 0.0066 to 0.0813 µg mL−1 (median = 0.0253 µg mL−1). Our results revealed a wide distribution in Mexico of Colletotrichum spp. isolates from mango sensitive to MBC fungicides and prochloraz. Data obtained in this study will serve as a source of comparison of the evolution of the sensitivity of Colletotrichum spp. populations obtained from mango to MBC and DMI fungicides in the future.

The contaminants of emerging concern (CECs) have been receiving global attention due to their worldwide presence in water bodies. The CECs could be originated from synthetic or natural sources, and they are not commonly monitored, although these substances are continuously reaching the aquatic environment. The main goal of this study was to determine the occurrence of some target CECs in Sao Paulo state surface water, once there is practically no information on the presence and concentration range of these substances at the studied sites. In addition, the present study aimed to assess adverse effects in the non-target fish embryo of Danio rerio (zebrafish) after exposure to surface water organic extract samples during 96 h using FET test. The CECs in surface water samples were determined by solid-phase extraction and liquid chromatography coupled by mass spectrometry. A 2-year study was assessed in 7 rivers and 3 reservoirs at Sao Paulo state, where 25 of the 30 analyzed substances were quantified, being caffeine the substance with the highest concentration range (5.5 ng L−1 to 69 μg L−1) and detected in 95% of analyzed samples, followed by bisphenol A (6.5–1300 ng L−1) and carbendazim (4.7–285 ng L−1), found in 50% and 85% of the analyzed samples, respectively. The chemical analysis and biological test were not performed in order to show a direct relationship between concentrations and observed effects on embryos; however, the combined approach can provide a better understanding of the adverse effects caused by mixtures of substances at relevant environmental concentrations. Regarding the adverse effects, it was observed that in the samples from sites with higher anthropogenic activity in the surroundings, there was also a higher mortality rate in organisms. At the Ribeirao Pires River and Sapucai-Guacu River, the mortality rate during the 2-year study was 21.6% and 9.3%, respectively. The morphological abnormality rates were higher at Ribeirao Grande (21.4%) and Ribeirao Pires (29.5%) Rivers. The obtained results aim to show that even in low concentrations (ng–μg L−1) the CECs can cause adverse effects on non-target species, and because of that, new chemical indicators would be important to monitor the water quality and protect the aquatic biota.

Cercospora leaf blight and purple seed stain are caused by Cercospora kikuchii and other Cercospora spp. The fungus can infect leaves and seeds on soybean, and the disease is controlled with fungicides. In Brazil, the intensive use of fungicides on soybean to control other diseases such as soybean rust has exerted resistance selection pressure on all fungal pathogens that attack in the crop. In this study, we evaluated the sensitivity of 56 Cercospora spp. soybean isolates collected during 9 crop seasons in 9 Brazilian states to quinone outside inhibitors (QoI) and methyl benzimidazole carbamate (MBC) fungicides using mycelial growth inhibition in amended media with a discriminatory dose of 10 μg/mL. We also analyzed single polymorphisms in the target genes cytb and β-tubulin using target genotyping by sequencing by Illumina short reads. Genome-local association was used to correlate the point mutations found in the coding sequence with in vitro assays results. For cytb gene, it was observed only the G143A mutation in 73% of the isolates. The mutation G143A was present in 97% of the isolates classified as resistant at least for two QoI fungicides. The mutation E198A was present in 71% of the isolates being significantly associated with the reduction control to MBC fungicides. Cross-resistance was observed into QoI (azoxystrobin, picoxystrobin, and pyraclostrobin) and MBC (carbendazim, and thiophanate-methyl) active ingredients fungicides. About 97% of Cercospora spp. isolates analyzed possessed the double mutations G143A and E198A and resulting in less sensitive to QoI and MBC fungicides, respectively. All isolates showed homozygous variation in the alternative allele in G143A and E198A mutations. Our results suggest that resistance of the Cercospora spp. from soybean to QoI and MBC fungicides has occurred since the 2008 cropping season in Brazil associated with G143A and E198A mutations.

The present work describes the development of a fully automated method based on online solid phase extraction (SPE)-liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous analysis of multiple classes of pesticides or metabolites in drinking water (DW), surface water (SW), and wastewater effluents (WWEs). The target list covers 111 pesticides or metabolites of various properties and families. LC-MS/MS and online SPE parameters were optimized with regard to the sorbent type, mobile phase composition, wash volume, and flowrate as well as the injection volume. The method showed good linearity in two concentration ranges with 97% and 94% of the coefficients (R2) being higher than 0.99 in the low concentration range (0.1-100 ng L-1) and high concentration range (100-2500 ng L-1), respectively. High sensitivity was observed with method quantification limits (MQLs) of 0.03-5.3 ng L-1, 0.06-17 ng L-1, and 0.08-21 ng L-1, for DW, SW, and WWE, respectively. The recoveries showed an accuracy of 94%, 91%, and 91% in the range of 70-130% for three matrices with satisfactory precision. The overall analysis time per sample was 30 min with minimum pretreatment. To the best of our knowledge, for the first time, 64 pesticides were identified by the high throughput online SPE-based method. The optimized method was used for WWE sample analysis, and 49 pesticides were detected in 12 WWE samples from an economically active city in China. Five pesticides were detected in all the samples, i.e. paclobutrazol, atrazine, diuron, acetamiprid, and triadimenol, and the highest median concentration was observed for carbendazim (324 ng L-1). The advantages of the proposed method over offline ones make it have broad prospects in high throughput and reliable analysis of pesticides in aquatic environments.

Bio-priming can also be viewed as a new technique of seed treatment using biological agents to stimulate germination of seed and growth of the plant and further protecting the seed from soil-and seed-borne pathogens. The present investigation was carried out in vitro conditions on seed germination and seedling vigour of few vegetable crops viz. tomato (Solanum lycopersicum L.), brinjal (Solanum melongena L.), onion (Allium cepa L.) and chilli (Capsicum annuum L.), during 2015 and 2016. The treatments comprised viz. T1: Non primed seeds (Control), T2: Seed treatment with Carbendazim 2.5g/kg seed, T3: Hydro-priming for 6 hrs, T4: Hydro-priming for 12 hrs, T5: Hydro-priming for 18 hrs, T6: Biopriming with Trichoderma viride for 6 hrs, T7: Biopriming with T.viride for 12 hrs, T8: Biopriming with T.viride for 18 hrs, T9: Biopriming with Trichoderma harzianum for 6 hrs, T10: Biopriming with T. harzianum for 12 hrs, T11: Biopriming with T. harzianum for 18 hrs, T12: Biopriming with Pseudomonas fluorescens for 6 hrs, T13: Biopriming with P. fluorescens for 12 hrs and T14: Biopriming with P. fluorescens for 18 hrs. The results revealed that maxiumum germination percentage (92.92, 90.77,83.00 and 86.33), seedling length (32.38 cm, 29.35 cm, 31.75 and 31.60 cm), seedling fresh weight (2.07 g, 4.01 g, 3.05 g and 2.04 g), seedling dry weight (0.42 g, 0.86 g, 0.62 g and 0.42 g) and seedling vigour index (3008.11, 2664.00, 2635.00 and 2728.00) were recorded in T10 (bio priming with T. harzianum for 12 hrs) in tomato (S. lycopersicum L.), brinjal (S. melongena L.), onion (A. cepa L.) and chilli (C. annuum L.), respectively. Thus, it indicated that priming of seeds of these crops with T. harzianum/P. fluorescens/ T. viride for 12 hrs was very effective with respect to their vegetative growth along with the quality yield.

Risks posed by pesticide residues in infant food urge for protection of the most vulnerable part of our population. In the current study a total of 54 samples of infant food (juice and puree) were collected on the Serbian market. Liquid chromatography-tandem mass spectrometry with electrospray ionization method detected 18 out of 69 analyzed pesticide active substances in 55.6% of the samples, most frequently carbendazim and acetamiprid. Domestic products as opposed to the imported ones showed a substantially higher proportion of positive (85% vs 38%) and noncompliant (10% vs 0%) samples, a number of pesticides detected (15 vs 8), the proportion of the samples with multiple residues (85% vs 15%), the maximum number of residues in an individual sample (7 vs 2). Risk assessment was performed for the present pesticide active substances, which was estimated to remain below the level of concern for both acute and chronic adverse health effects.

Soil-borne pathogens and weeds could synergistically affect vegetable growth and result in serious losses. The investigation of antagonistic metabolites from a marine-derived entomopathogenic fungus, Beauveria felina, obtained polyhydroxy steroid (1), tricyclic diterpenoid (2), isaridin (3), and destruxin cyclodepsipeptides (4-6). The structures and absolute configurations of new 1-3 were elucidated by extensive spectroscopic and X-ray crystallographic analyses, as well as electronic circular dichroism (ECD) calculations. Compounds 1 and 2 showed antifungal activities against carbendazim-resistant strains of Botrytis cinerea, with the minimum inhibitory concentration (MIC) values ranging from 16 to 32 μg/mL, which were significantly better than those of carbendazim (MIC = 256 μg/mL). Compound 5 exhibited significant antagonistic activity against the radicle growth of Amaranthus retroflexus seedlings, which was almost identical to that of the positive control (2,4-dichlorophenoxyacetic acid). The structure-activity differences of 4-6 suggested that the Cl atom in HMPA1 and β-Me in Pro2 should be the key factors to their herbicidal activities. Besides, compounds 3-6 showed moderate nematicidal activities against Meloidogyne incognita. These antagonistic effects of 1-6 were first reported and further revealed the synergistically antagonistic potential of B. felina to be developed into the biopesticide.

We have shown, the outcome of antifungal activity of phenazine derivatives which is produced by fluorescent pseudomonads (FPs) for the control of sheath blight of rice. A total of 50 fluorescent pseudomonads (FPs) were isolated from rice rhizosphere. Off which, 36 FPs exhibited antagonistic activity against Rhizoctonia solani, Macrophomina phaseolina, Fusarium oxysporum, Alternaria alternata and Sclerotium rolfsii up to 70-80% compared to control by dual culture method. BOX-PCR analyses of antagonistic isolates indicated that two phylogenetic group, where group I consisted of 28 isolates and eight isolates belongs to group II. Among 36 FPs, a total of 10 FPs revealed that the presence of phenazine derivatives on thin layer chromatography (TLC), which is coincided with that of authentic phenazine with Rf value 0.57. Similar to TLC analysis, antibiotic encoding gene phenazine-1-carboxamide (PCN) was detected in 10 FPs by PCR analysis with respective primer. Among, PCN detected isolates of FPs, a significant biocontrol potential possessing isolate designated as VSMKU1 and it was showed prominent antifungal activity against R. solani and other tested fungal pathogens. Hence, the isolate VSMKU1 was selected for further studies. The selected isolate VSMKU1 was identified as Pseudomonas aeruginosa by 16S rDNA sequence analysis. The antifungal metabolite phenazine like compound produced by VSMKU1 was confirmed by UV, FT-IR and HPLC analysis. The phenazine compound from VSMKU1 significantly arrest the growth of R. solani compared to carbendazim by well diffusion method. The detached leaf assay showed remarkable inhibition of lesion height 80 to 85% by the treatments of culture (VSMKU1), cell free culure filtrate and phenazine like compound compared to control and other treatments was observed in detached leaves of rice. These results emphasized that VSMKU1 isolate can be used as an alternative potential biocontrol agent against sheath blight of rice, instead of using commercial fungicide such as validamycin and carbendazim which cause environmental pollution and health hazards.

Because of diverse adverse effects of carbendazim (CBZ) upon the human body and environment, a simple method for the determination of this molecule is urgently needed. Therefore, a method based on ...

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum, is a devastating fungal disease in oilseed rape. Chemical control with fungicides is the primary method of controlling this disease. Metconazole is a demethylation inhibitor fungicide that has high activity against mycelial growth of phytopathogenic fungi. In this study, 119 isolates of S. sclerotiorum were collected from oilseed rape fields in different regions of Henan Province, China, in 2015 and 2016 prior to exposure to metconazole. The sensitivities of the S. sclerotiorum isolates to a range of concentrations of metconazole were determined based on mycelial growth inhibition in 90 cm Petri-dishes containing Potato dextrose Agar to which the fungicide was added. The 50% inhibition of mycelial growth (EC50) values of the S. sclerotiorum population to metconazole ranged from 0.0469 to 0.3912 µg mL− 1 and the mean EC50 value was 0.1875 ± 0.0058 µg mL− 1 (mean ± standard error). The frequency distribution range curve was unimodal with a narrow range. Spearman’s rho (ρ) for the log10 of the EC50 values of 22 isolates of S. sclerotiorum between metconazole and carbendazim, dimethachlone, boscalid, fluazinam, and fludioxonil showed that there was no cross-resistance between metconazole and the other test fungicides. The results from this study provide important information for the control of Sclerotinia stem rot and sensitivity monitoring of S. sclerotiorum to metconazole in this area.

In the present study, a total of 95 Botrytis cinerea single-spore strains collected from different hosts in Shaanxi Province of China were characterized for their sensitivity to the sterol demethylation inhibitor fungicide flusilazole. The effective concentration for 50% inhibition of mycelial growth (EC50) of flusilazole ranged from 0.021 to 0.372 µg/ml, with an average value of 0.093 µg/ml. Cross-resistance between flusilazole and commonly used fungicides was not detected, and no flusilazole-resistant mutants were induced. Both on detached strawberry leaves and in greenhouse experiments, flusilazole was more effective than the commonly used fungicide carbendazim at reducing gray mold. After culture on PDA plates or detached strawberry leaves, no difference in sclerotia production or pathogenicity was detected between two strains, WG12 (most sensitive to flusilazole) and MX18 (least sensitive to flusilazole). After treatment with flusilazole, however, the two strains lost the ability to produce sclerotia, and oxalic acid and ergosterol contents in mycelium decreased. Interestingly, the inhibition rate of ergosterol content in MX18 was significantly lower than that in WG12. Expression of Cyp51, BcatrD, and Bcmfs1 genes all increased after treatment with flusilazole, especially the Cyp51 and BcatrD genes. However, the expression of Cyp51 gene or BcatrD gene in WG12 and MX18 were significantly different from each other after treatment with flusilazole. In addition, no point mutations in Cyp51 gene were found in MX18. These data suggest flusilazole is a promising fungicide for resistance management of gray mold and also provided novel insights into understanding the resistance mechanism of flusilazole against plant pathogens.

An analytical method for the accurate determination of carbamate pesticides, carbaryl, carbendazim, and carbofuran, in vegetables was established based on the isotope dilution liquid chromatography/tandem mass spectrometry (ID-LC/MS/MS) technique as a candidate reference method. For sample clean-up, a Carb/LC-NH2 solid-phase extraction (SPE) cartridge was employed to ensure an efficient recovery and color removal from the vegetable samples. Isotope analogues, 13C6-carbaryl, carbendazim-d4, and 13C6-carbofuran, were used as internal standards, respectively. This method was validated by measuring samples fortified with the three target analytes into vegetable samples and prepared homogeneous Korean cabbage powder samples in this laboratory. All results of the measured values agreed with the fortified values within their uncertainties. Repeatability and reproducibility of the method were both approximately 2%. As commercially available CRM contains only carbendazim, its content was analyzed by the developed method. The measured value of carbendazim in CRM was (0.0724 ± 0.0013) mg/kg agreeing with the certified value (0.074 ± 0.004) mg/kg. These validation data indicate the developed ID-LC/MS/MS method is reproducible, repeatable and reliable to be as a candidate reference method for accurate determination of carbamate pesticides in vegetables.

In the present study, chitosan nanoparticles were synthesized from four different fungal sources viz., Fusarium oxysporum, Metarhizium anisopliae, Beauveria bassiana and Trichoderma viride and from one commercial source of chitosan.UV–VIS spectroscopy study of synthesized nanoparticles showed absorption peaks at the range of 310.02 to 342.00 nm. Functional groups of chitosan nanoparticle as OH, N–H, C–H, C=O, C–O, C–N and P=O were confirmed by FTIR study. Surface properties of the synthesized nanoparticles were found to be positively charged and were found stable in nature. Electron microscopy study showed that synthesized nanoparticles are nearly spherical in shape. DLS analysis showed the average size of the chitosan nanoparticles synthesized from F. oxysporum, M. anisopliae, B. bassiana, T. viride and commercial products as 273.20, 172.50, 78.36, 89.03 and 300.10 nm respectively. Compatibility study of chitosan nanoparticles with T. asperellum showed highly compatible reaction with chitosan nanoparticles at the tested concentration. In vitro assay against Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii showed combination of T. asperellum and chitosan nanoparticle as superior in suppression of mycelial growth of the pathogens as compared to Trichoderma alone and carbendazim @0.1%.

The present study was aimed to isolate active constituents from Blumea axillaris (Lam.) DC (Asteraceae) against phytopathogenic fungi. Bioactivity guided fractionation of the successive n-hexane, chloroform and methanol extract led to the isolation of the monoterpene ester (4 R,5S)-4-hydroxy-7-tigloyloxycarvotanacetone (1). The compound 1 was converted into acetyl derivative (2). The acetyl derivative (2) and the parent compound 1 were tested again phytopathogenic fungi by using mycelial inhibition and minimal inhibitory concentration values were found out by the broth microdilution method. The acetyl derivative (2) showed the highest antifungal activity against Rhizoctonia solani and Aspergillus niger. Based upon in vitro results, compound 1 was tested against Fusarium oxyporum (wilting disease) and compound 2 was tested against R. solani (leaf blight disease) in vivo using the foliar spray method. Both compounds had no phytotoxicity and also in silico docking study showed that both compounds were binding similarly as commercial fungicide carbendazim.

In this study, the fungicidal activities of the fungicides azoxystrobin, difenoconazole + propiconazole, carbendazim, flutriafol, fluopyram + tebuconazole, mancozeb and thiophanate-methyl against rice blast and dirty panicle pathogens were evaluated under laboratory and field conditions. Mancozeb exhibited the highest level of fungicidal activity against the blast pathogen Pyricularia oryzae, with an EC50 value of 0.25 parts per million (ppm). The combination of two fungicides, fluopyram + tebuconazole, showed the strongest fungicidal effect against Bipolaris oryzae and Curvularia lunata, with EC50 values of 0.587 ppm and 0.435 ppm, respectively. Meanwhile, carbendazim and flutriafol demonstrated the best level of fungicidal activity against Fusarium incarnatum, with the lowest EC50 values of 0.211 ppm and 0.214 ppm, respectively. The results showed that the fungicides, triazole and strobilurin, had significant effects against rice blast and dirty panicle diseases. The combination of fluopyram + tebuconazole, when applied twice, was the most effective in reducing dirty panicle disease by up to 60% and increasing rice yield by 29% more than the untreated control. Fluopyram + tebuconazole, difenoconazole + propiconazole, flutriafol and azoxystrobin achieved stronger fungicidal activity against rice blast disease, reducing its severity by 32–33% when applied twice by foliar spraying. However, carbendazim, mancozeb and thiophanate-methyl had low to moderate fungicidal activity against both rice diseases in this study.

The increasing prevalence of environmental pollutants such as pesticides is a major global problem that affects living organisms. Exposure to environmental pollutants remains a major source of health risk throughout the world. The potential health benefits of various medicinal plants and natural products in relation to protect various diseases are currently receiving considerable attention. A current approach is to develop a new biological compound from natural products that inhibits pain. Ethnopharmacological surveys have been found to be one of the most reliable tools for the discovery of the natural and semi-synthetic drug. The present study was performed to investigate the hematological and biochemical changes induced by carbendazim (CBZ) and the potential protective effect of seeds extracts of Nigella sativa (NSSE) and Foeniculum vulgare (FVSE) against CBZ toxicity in male mice. Mice were distributed into 6 groups. Mice of group 1 were served as control. Group 2 was exposed to CBZ. Group 3 was supplemented with NSSE and exposed to CBZ. Group 4 was treated with FVSE and CBZ. Normal mice of group 5 and 6 were subjected to NSSE and FVSE respectively. Body weight gain was significantly decreased in mice of group 2. In mice of group 2, significant declines of RBC, HB, Hct, WBC, total protein, FSH, LH, testosterone, T4, T3, CAT and SOD were observed. Moreover, the levels of ALT, AST, ALP, total bilirubin, creatinine, BUN, uric acid, glucose, cholesterol, CK, LDH, MDA and GSH were significantly enhanced. Treatment with NSSE and FVSE showed attenuation effects against CBZ induced hematological and biochemical changes. The results suggest that the attenuation effects of NSSE and FVSE attributed to their antioxidant properties.

A study of the genotoxic activity of fungicide carbendazim was carried out. It was demonstrated that the tested technical grade active ingredient of carbendazim in the dose range of 125–2000 mg/kg body weight induced a statistically significant dose-dependent increase in the frequency of micronucleated polychromatic (PCE) and normochromatic erythrocytes (NCE) in the bone marrow of CD-1 mice. After administration of carbendazim at doses of 250–2000 mg/kg body weight, the accumulation of cells containing two or more micronuclei was observed. According to the results of comet assay, there was no increase in the level of breaks and alkali-labile sites in the DNA of the mouse bone marrow and liver cells after administration of technical grade carbendazim at doses up to 2000 mg/kg body weight. It is therefore suggested that the observed accumulation of cells with micronuclei is due to the aneugenic effect of carbendazim. In addition, a dose-dependent increase in the proportion of macrocytes/gigantocytes and dumbbell-shaped cells with constrictions among PCEs and NCEs was observed, which might be indicative of the inhibitory effect of the test pesticide on cytokinesis. The accumulation of polychromatophiles with pyknotic nuclei after exposure to carbendazim is probably caused by the distortion of nuclear extrusion. A possible general mechanism underlying the impairment of karyokinesis, cytokinesis, and nuclear extrusion in mammalian bone marrow erythroid cells is discussed.

Background Integrating fungal biocontrol agents into crop protection programs dominated by synthetic pesticides is an important first step towards developing an IPM program, however their successful integration relies on an understanding of how their performance may be impacted by the remaining agrochemicals deployed for managing other pests and diseases. In this study we tested ten formulated pesticides used in macadamia production at different concentrations to determine their effects on the germination, mycelial growth and sporulation of Metarhizium anisopliae and Beauveria bassiana in vitro. Further tests with laboratory grade actives of the non‐compatible pesticides were conducted to determine whether any antagonistic effects were caused by the active constituent or by formulation additives. Results At their registered concentrations, formulated trichlorfon, acephate and indoxacarb were compatible with M. anisopliae, whereas B. bassiana showed compatibility with formulated trichlorfon, acephate, indoxacarb, sulfoxaflor and spinetoram. Bioassays using laboratory grade active constituents indicated that the adverse impact of formulated beta‐cyfluthrin on both fungal species and that of formulated methidathion on B. bassiana is probably due to components of the emulsifiable concentrate formulations rather than their active constituents. Diazinon was the only insecticidal active that showed high toxicity to both fungal species. The two fungicides, carbendazim and pyraclostrobin, were toxic to both fungal species at all tested concentrations. Conclusion Our results identify which pesticides used on macadamias in Australia are compatible and incompatible with entomopathogenic fungi. Future studies on pesticides degradation rates will help define the spray intervals required to eliminate these adverse effects.

Sheath blight is one of the most important diseases of rice worldwide causing serious yield losses. The study was conducted to determine the best and most eco-friendly combination of different IDM practices for the management of the disease under field conditions. Research findings over two crop seasons, suggests that IDM practices (treatment T3) which included seed treatment with carbendazim (2 g/kg) along with one foliar application of carbendazim (@1 g/m2) 7 days before uprooting the seedlings + application of DAP @ 108 g/10 m2 nursery area (to supply 20 kg nitrogen and 50 kg phosphorus per hectare) and MOP @ 85 g/10 m2 nursery area (to supply 50 kg potassium/ha) before sowing + application of 120N:60P:40K:25Zn (100% recommended dose). Additional 25% N was applied at booting to make the plants more prone to the diseases) + one blanket application of granular insecticide (cartap hydrochloride 4% G @ 18.75 kg/ha) at 15 DAT + two foliar sprays of propiconazole @ 0.1% first at booting stage and second after booting stage, if disease appears was proved most effective and consistent in reducing the severity (46.21%) and incidence (39.72%) of sheath blight disease. The treatment also increased the grain yield up to 10.83%. This shows that the IDM practices have significant effect on disease management and also in improving the yield.

Green mold decay is considered as the main cause of postharvest loss in citrus fruits. With regard to safety concerns, this study was done to evaluate the efficacy of shellac as an edible coating and cinnamon essential oil (CEO) as alternative to synthetic fungicides to maintain quality of ‘Thomson navel’ oranges (Citrus sinensis L. Osbeck) inoculated with Penicillium digitatum spores. Fruit treated with distilled water (control), 1.5% ethanol (ET), 10% shellac (SH), commercial wax (CW), 0.5% commercial fungicide (Carbendazim), CF + CW, 0.3%, 0.4%, 0.5% and 0.6% CEO, 0.3%, 0.4%, 0.5% and 0.6% CEO + SH, SH enriched with 0.3%, 0.4%, 0.5% and 0.6% CEO (CEOSH) stored at 5 °C for up to 21 days. Fruit decay, weight loss, firmness, ascorbic acid were evaluated at 7 days interval, but scanning electron microscopy (SEM) images and sensory quality were evaluated at the end of storage. Shellac coating (10%) enriched with 0.5% CEO reduced weight loss by 52%, and firmness loss by 38%. The results showed that the incorporation of EOs into shellac could be a suitable treatment for maintenance of citrus fruit quality.

Green synthesis of silver nano-particles (AgNPs) from silver nitrate was carried out using purple-colored rice leaves' extracts containing higher phenols, anthocyanins, and flavonoids. The efficacy of synthesized AgNPs was tested against rice diseases and investigation was carried out to check negative effect of AgNPs on soil microbes. Substantial reduction of total anthocyanins, total phenols, and total flavonoids was observed in reaction mixture during AgNP formation indicating the role of secondary metabolites on AgNP formation and stabilization. Scanning electron microscopy coupled with energy-dispersive spectroscopic images and FTIR spectral analysis of AgNPs confirmed the presence of elemental silver encapped by biomolecules. The optimized reaction parameters for synthesis of AgNPs from silver nitrate were (a) 48 h of incubation, (b) 9:1 (v/v) 1 mM AgNO3:plant extract, and (c) room temperature at 20-30 °C. Zeta potential and hydrodynamic particle sizes of synthesized AgNPs were ranged between - 16.61 to - 29.45 mV and 36-107 nm, respectively, at different time of incubation. AgNPs could control effectively Rhizoctonia solani and Xanthomonas oryzae pv. Oryzae and Helminthosporium oryzae. AgNPs at higher concentration could cause negative effect on microbial biomass carbon and soil enzymes for distant future. But the negative effects of AgNP solution (10% of 1 mM AgNPs) were comparable to commercial fungicide, carbendazim. The synthesized AgNPs with desirable characters were effective against a number of disease-causing pathogens in rice, and it can be recommended as broad-spectrum pesticide.

Fungi have been proved as promising and prolific sources of functional secondary metabolites with potent agricultural applications. In this study, 14 xanthone derivatives (1-14), including six new ones, versicones I-N (1-4, 7, 11), and a biogenetically related derivative (15), were isolated from the alga-derived fungus Aspergillus versicolor D5. Their structures were elucidated by comprehensive spectroscopic methods. Versicone L (4) exhibited a broad antifungal spectrum and prominent inhibitory effects on Botrytis cinerea at a minimum inhibitory concentration (MIC) of 152 μM, 7-fold stronger than that of the positive control, carbendazim (MIC = 1.05 × 103 μM). Dihydrosterigmatocystin (13) showed strong antifungal activity toward B. cinerea at MIC = 38.3 μM, almost 30-fold stronger than that of carbendazim. Meanwhile, 13 exhibited potent herbicidal activity toward Amaranthus retroflexus L. with an MIC of 24.5 μM, approximately 4-fold stronger than that of the positive control, glyphosate (MIC = 94.7 μM). Additionally, 13 also displayed remarkable activity against other weeds belonging to Amaranth sp. Analysis of the structure-herbicidal activity relationship indicated that the bifuranic ring played an important role in xanthone phytotoxicity and the presence of a double bond in the furan ring could decrease phytotoxicity. This study indicated that xanthones can be served as promising candidates for lead compounds of agrochemicals.

Fusarium graminearum is the primary causal agent of Fusarium head blight (FHB) of wheat. The phenylpyrrole fungicide fludioxonil is not currently registered for the management of FHB in China. The current study assessed the fludioxonil sensitivity of a total of 53 F. graminearum isolates collected from the six most important wheat-growing provinces of China during 2018 and 2019. The baseline fludioxonil sensitivity distribution indicated that all of the isolates were sensitive, exhibiting a unimodal cure with a mean effective concentration for 50% inhibition value of 0.13 ± 0.12 μg/ml (standard deviation). Five fludioxonil-resistant mutants were subsequently induced by exposure to fludioxonil under laboratory conditions. Ten successive rounds of subculture in the absence of the selection pressure indicated that the mutation was stably inherited. However, the fludioxonil-resistant mutants were found to have reduced pathogenicity, higher glycerol accumulation, and higher osmotic sensitivity than the parental wild-type isolates, indicating that there was a fitness cost associated with fludioxonil resistance. In addition, the study also found a positive cross resistance between fludioxonil, procymidone, and iprodione, but not with other fungicides such as boscalid, carbendazim, tebuconazole, and fluazinam. Sequence analysis of four candidate target genes (FgOs1, FgOs2, FgOs4, and FgOs5) revealed that the HBXT2R mutant contained two point mutations that resulted in amino acid changes at K223T and K415R in its FgOs1 protein, and one point mutation at residue 520 of its FgOs5 protein that resulted in a premature stop codon. Similarly, the three other mutants contained point mutations that resulted in changes at the K192R, K293R, and K411R residues of the FgOs5 protein but none in the FgOs2 and FgOs4 genes. However, it is important to point out that the FgOs2 and FgOs4 expression of all the fludioxonil-resistant mutants was significantly (P < 0.05) downregulated compared with the sensitive isolates (except for the SQ1-2 isolate). It was also found that one of the resistant mutants did not have changes in any of the sequenced target genes, indicating that an alternative mechanism could also lead to fludioxonil resistance.

Sheath blight of rice is one of the most damaging diseases of rice worldwide causing serious yield losses. Several fungicide molecules with different mode of actions are available in the market. The present study was undertaken to find the best and most economical combination of fungicides in managing this disease. Hence, an experiment was conducted to evaluate the efficacy and economics of flusilazole 12.5% + carbendazim 25% SC, difenoconazole 11.4% + azoxystrobin 18.2% SC, tebuconazole 18.3% + azoxystrobin 11% SC, tricyclazole 18% + mancozeb 62% WP, zineb 68% + hexaconazole 4% WP, trifloxystrobin 25% + tebuconazole 50% WG, mancozeb 50% + carbendazim 25% WS and epoxiconazole 62.5 g/l + fluxapyroxad 62.5 g/l EC against sheath blight of rice under the field conditions. Results of the present study revealed that, out of the different fungicidal combinations, two foliar sprays of difenoconazole 11.4% + azoxystrobin 18.2% SC @ 1 ml/lit was found to be most economic that provided best incremental net benefit ratio of 6.17:1.00. It was followed by two foliar sprays of epoxiconazole 62.5 g/l + fluxapyroxad 62.5 g/l EC @ 1.5 ml/l, tebuconazole 18.3% w/w + azoxystrobin 11% SC, zineb 68% + hexaconazole 4% WP and trifloxystrobin 25% + tebuconazole 50% WG.

Charcoal rot disease of soybean caused by Macrophomina phaseolina is a serious problem in most of the soybean growing area of Madhya Pradesh. In this study, seven plant extracts viz., leaves of Azadirachta indica, Citrus limon, Polyalthia longifolia, Parthenium hysterophorus and Ricinus communis, bulb of Allium sativum and Allium cepa and eight fungicides viz., Captan (0.25%), Mancozeb (0.25%), Carbendazim + Mancozeb (25%), Thiophanate Methyl (0.1%), Pyraclostrobin (0.2%), Carbendazim (0.1%) and Blue copper (0.3% ) were evaluated for an effective management of charcoal rot of soybean caused by M. phaseolina (Tassi) Goid under in vitro and in vivo condition. Among plant extracts, garlic clove extract was found most effective showing 77.3 % growth inhibition and poor microslerotia formation of M. phaseolina by 77.3 % followed by parthenium leaf extract (75.2% inhibition) at 15 % concentration. Two soil drenching of garlic clove extracts @ 15.0 % concentration also found most effective for the management of disease under field condition recorded minimum disease incidence (13.5%) and highest yield (14.6q/ha). Among fungicides, Carbendazim (0.1%) and Thiophanate Methyl (0.1%) showed 100 % inhibition of radial growth and microsclerotia production of M phaseolina under in vitro condition. Two soil drenching of Carbendazim @ 0.1% found to be most effective for the management of charcoal rot of soybean under field condition showing minimum disease incidence (5.36%) and producing highest yield (16.0 q/ha) followed by Thiophanate Methyl. These results suggested that the toxic effect of Carbendazim and Thiophanate Methyl and A. sativum inhibited maximum mycelium growth in vitro and provide management of charcoal rot disease under field conditions.

The purpose of this study was to evaluate the capability of Aspergillus tubingensis and Talaromyces islandicus to solubilize inorganic phosphorus sources, their activity under abiotic stress, and the enhancement of P availability in soils and plant growth. The P-solubilizing capability and acidification mechanism of the strains were assessed in vitro using tricalcium phosphate and rock phosphate. Independent assays were conducted with rock phosphate under NaCl and fungicides carbendazim, chlorothalonil, and propamocarb hydrochloride using a factorial design. Thereafter, the effects of fungal inoculations in rock phosphate–amended soil and P nutrition of Zea mays were assessed in a greenhouse experiment. Both fungi solubilized P in vitro via acidification through the exudation of acetic, citric, lactic, malic, quinic, and succinic acids. The P-solubilizing efficiency of A. tubingensis was maintained above 97.5% under 0.5 to 3.0% NaCl, up to 28.7% in the treatment with carbendazim, up to 5.3% with chlorothalonil, and above 96.5% with propamocarb hydrochloride; while T. islandicus efficiency decreased to 45.2% in a NaCl concentration-dependent trend, and maintained it above 80% in the fungicide treatments. The inoculation with A. tubingensis increased the available P in the amended soil by up to 65% after 30 days and resulted in 87% higher foliar P content, 111% greater plant height, and 25% greater dry weight of maize shoots. Similarly, T. islandicus contributed to these parameters in 55, 67, 90, and 17%, respectively. These findings suggest their potential as qualified phosphorus solubilizing microorganisms to develop novel and sustainable approaches for P fertilization in agriculture.

Wheat stem rust caused by Puccinia graminis f. sp. tritici is an important wheat disease with sudden and devastating characteristics. The appearance and spread of new P. graminis f. sp. tritici races (Ug99, TKTTF, and TTTTF) have once again renewed the interest in the prevention and control of wheat stem rust. Fungicides can effectively control the epidemics of this disease in a short period of time. However, the fungal pathogen is prone to developing resistance. Therefore, we collected 89 isolates of P. graminis f. sp. tritici from four provinces in China and used the spore germination method to test the sensitivity of the isolates to fungicide triadimefon. Seven relatively triadimefon-sensitive isolates and six relatively triadimefon-resistant isolates were further tested for sensitivity to fungicides carbendazim, mancozeb, thiophanate-methyl, and kresoxim-methyl. The results showed that the mean concentration for 50% of maximal effect of the isolates to triadimefon was 16.14 mg·liter−1, and the mean resistance factor was 4.48. Only 29 isolates were resistant to triadimefon in which 27 isolates had low levels of resistance and 2 isolates had moderate levels of resistance. However, most of the 89 isolates had no resistance to triadimefon. There was a positive correlation between resistance to triadimefon and carbendazim, but there was no cross-resistance between triadimefon resistance with thiophanate-methyl or kresoxim-methyl resistance. This study provides valuable information for managing fungicide resistant isolates of P. graminis f. sp. tritici.

Introduction. From the evaluation of the positions of residual quantities of pesticides, the hygienic safety assessment of imported food products is most relevant under the current agreement on a free trade zone between the EAEU and the Socialist Republic of Vietnam, as well as in connection with a large list of chemical plant protection products used during the vegetation of crops. The purpose of the study. Rating the level of residual quantities of pesticides in samples of selected food products produced in Vietnam is the purpose of the study. Research objectives. To quantify 42 active ingredients of pesticides (and their metabolites) in samples of rice grain, dragon fruit (pitahaya), avocado, mango and banana (fresh and dried) are the research objectives. Material and methods. Multicomponent and individual methods based on LC and GC with MS-detection and FLD were used to identify active ingredients of pesticides. The sample preparation procedure with QuEChERS technology was used for the multi residues determination of the 40 compounds. The determination of avermectins (abamectin and emamectin benzoate) was carried out after the conversion of substances into fluorogenic derivatives. Results. Residual amounts of tebuconazole (rice - 0.53 ppm, mangoes - 0.26 ppm, avocado - 0.15 ppm), fludioxonil (rice - 0.017 ppm), imidacloprid (dried mangoes - 0.01 ppm), carbendazim (dried mangoes - 0.011 ppm) were identified in single samples No residual avermectins have been identified. The content of substances in the analyzed samples of dragon fruit (pitahaya) did not exceed 0.01 ppm. Discussion. The results obtained in the work are compared with the data of the annual pan-European and national monitoring, information on which is publicly available (2017), including imported goods, according to which rice and dragon fruits as objects of monitoring are given increased attention. Conclusions. The safety of certain types of food products imported from Vietnam is shown by the content of residual quantities of pesticides.

Wilt (Fusarium oxysporum f. sp. lycopersici) of tomato is one of the major factors for low productivity of the crop. Bio-control agents along with fungicides were evaluated in various modes of applications to develop an efficient wilt management module. Tomato seedling roots dipped in fungicide carbendazim and bioformulation of Trichoderma harzianum and T. viride separately provided the lowest wilt incidence with the highest shoot and root lengths, plant dry weight, number and weight of fruits with statistically similar performance. Both T. harzianum and T. viride based soil application formulations showed similar performances. Further based on these results, seedling root dip and soil application of T. harzianum and T. viride separately and in combinations along with fungicides and Bacillus subtilis were evaluated. Combined application of PBP10G (T. harzianum) in soil and seedling root dip in a suspension of Pusa 5SD (T. harzianum) + fungicide carboxin 37.5% + tetramethyl thiuram disulfide 37.5% (Vitavax power) provided the highest shoot length, fruit number and weight along with the lowest wilt incidence. The wilt incidence observed in the treatment was alike to that of T. harzianum (PBP10G) × T. viride (Pusa 5SD) + Vitavax power and T. viride (PBP10G) × T. harzianum (Pusa 5SD) + Vitavax power. Thus, the application of Trichoderma based formulations with fungicide Vitavax power not only reduced the incidence of wilt, but also enhanced the growth of plants and fruit yield. The module developed is entirely new and proved effective for tomato wilt management.

Fusarium head blight (FHB), caused by Fusarium graminearum species complex (FGSC) is one of the most destructive wheat diseases worldwide. F. graminearum is the dominant pathogen of FHB in Henan Province. Epoxiconazole is a demethylation inhibitor (DMI) fungicide, and is the main fungicide for controlling FHB in China. In this study, 118 F. graminearum isolates were collected from different regions of Henan Province in 2016 and 2017. The sensitivity of these F. graminearum isolates to epoxiconazole was determined by the method of mycelial growth inhibition. The results showed that the EC50 values range of epoxiconazole to the tested F. graminearum isolates was 0.0196 to 0.5708 μg ml−1, and the mean EC50 value was 0.2178 ± 0.1178 μg ml−1 (mean ± SD). The sensitivity frequency distribution of these F. graminearum isolates to epoxiconazole was a continuous single peak curve, and no isolates resistant to epoxiconazole were found in Henan Province, China. The field control efficacy of epoxiconazole against F. graminearum was evaluated and the results showed that epoxiconazole had a significantly higher control efficacy on FHB, compared to the control fungicide carbendazim. The thousand grain weight of all the fungicide treatments was significantly higher than water control (without fungicide). This study showed that the DMI fungicide epoxiconazole has high activity and field control efficacy on F. graminearum, and can effectively control the prevalence of FHB.

It is crucial to develop practical procedures for the control and reduction of pesticide residues in oil productions from farm to dining table. In this study, the dissipation behaviors of typical fungicide from rapeseed to oil production were studied to reveal relationship among spraying stage, application dosage, household oil processing stage, and pesticide residues. In the field trials, rape plants were sprayed with carbendazim at three different dosages during flowering period. Transfer assessment of carbendazim residues from rapeseed to oil production during household oil processing via different press techniques was determined using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). The recoveries of carbendazim in rapeseed samples, meals after squeezing samples, and rapeseed oil samples ranged from 82.5% to 93.6% with relative standard deviations (RSDs) ＜5.2%. The validation results illustrated that the methods were reliable and sensitive. The average processing factor (PF) during household oil processing via hot press technique and cold press technique was 0.15 and 0.51, respectively. This study demonstrated that household oil processing could significantly reduce the pesticide residues, especially by hot press technique.

Agri-vitality of benzimidazoles and 1,2,4-triazoles against ergosterol and β-Tubulin prompted and “lead hybridization” based novel series of benzimidazolyl-1,2,4-triazoles (1–32) were designed as twin-enzyme targeted inhibitors. In silico computational tools viz. molecular docking, Lipinski parameters, Frontier molecular orbital approach and Toxicity analysis screened three benzimidazoly-1,2,4-triazoles out of 32 designed molecules, which were synthesised by multistep protocol and characterized by spectroscopic techniques. Antimycotic activity against F. verticillioides, D. oryzae, C. lunata and F. fujikuroi indicated deca fold enhanced potency of all the synthesised compounds, than the standard commercial benzimidazole fungicide, carbendazim. Compounds 8 exhibited ED50 values lower than triazole fungicide, propiconazole. Remarkably, compound 8 inflicted the most promising activity against all the test fungi with ED50 value ranging from 16 to 21 μg/ml better than the standard commercial fungicides used (Tilt: 20–25 μg/ml and Carbendazim:150–230 μg/ml). Ultra microscopic details revealed compound 8 not only caused aberrant distortions resulting in collapsed hyphae but also efficiently shrunken the spores resulting in reproduction inhibition, as possible cause of fungal growth inhibition.

Gray mold caused by Botrytis cinerea is a fungal disease that critically threatens agricultural production, and carbendazim was the first fungicide used to control B. cinerea. However, B. cinerea developed serious resistance to carbendazim, and this fungicide has thus rarely been used in the past decade in China. Due to the extended discontinuation of carbendazim use, the evolution of the resistance of B. cinerea to carbendazim in recent years is unclear, and whether carbendazim can effectively control gray mold is largely unknown. Therefore, this study determined the sensitivity of 407 B. cinerea isolates collected from 2014 to 2018 to carbendazim and the ability of carbendazim to control gray mold in the field. The results showed that the frequency of B. cinerea isolates resistant to carbendazim remained above 95%. Three different mutation types responsible for the resistance of B. cinerea to carbendazim were identified at codon 198 in the β-tubulin gene sequence: E198V (changed from GAG to GTG), E198A (changed from GAG to GCG), and E198K (changed from GAG to AAG). Over the last 5 years, E198V was the major mutation. However, an analysis of its evolution revealed that the percentage of the E198V mutation declined after 2017 to 56.5% in 2018. In addition, the proportion of isolates with the E198K mutation decreased over time, and no isolates with this mutation were found in either 2017 or 2018. The proportion of the E198A mutation increased over the 5-year test period to reach 43.5% in 2018. Furthermore, three greenhouse experiments demonstrated that carbendazim has lost its ability to control gray mold. We attribute the above findings to our results showing that the carbendazim-resistant isolates had no fitness penalties compared with the carbendazim-sensitive isolates for sporulation and mycelial growth. In particular, the E198A mutant isolates exhibited a strong ability to sporulate, suggesting that the E198A mutation might become dominant in the future. Interestingly, the results showed that carbendazim-sensitive isolates could be easily controlled by four conventional fungicides, namely boscalid, procymidone, iprodione, and pyrimethanil, with mean EC50 values of 0.71 ± 0.2 mg liter−1, 1.33 ± 0.39 mg liter−1, 0.59 ± 0.33 mg liter−1, and 6.02 ± 3.02 mg liter−1, respectively. In conclusion, carbendazim has lost its application value and is ineffective for the control of gray mold.