Molybdenum and cadmium co-induce apoptosis and ferroptosis through inhibiting Nrf2 signaling pathway in duck (Anas platyrhyncha) testes.

Cadmium (Cd) and high molybdenum (Mo) are injurious to the body. Previous research has substantiated that Cd and Mo exposure caused testicular injury of ducks, but concrete mechanism is not fully clarified. To further survey the toxicity of co-exposure to Cd and Mo in testis, 40 healthy 8-day-old Shaoxing ducks (Anas platyrhyncha) were stochasticly distributed to 4 groups and raised with basic diet embracing Cd (4 mg/kg Cd) or Mo (100 mg/kg Mo) or both. At the 16th wk, testis tissues were gathered. The characteristic ultrastructural changes related to apoptosis and ferroptosis were observed in Mo or Cd or both groups. Besides, Mo or Cd or both repressed nuclear factor erythroid 2-related factor 2 (Nrf2) pathway via decreasing Nrf2, Heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQO1), Glutamate-cysteine ligase catalytic subunit (GCLC) and Glutamate-cysteine ligase modifier subunit (GCLM) mRNA expression of and Nrf2 protein expression, then stimulated apoptosis by elevating Bcl-2 antagonist/killer-1 (Bak-1), Bcl-2-associated X-protein (Bax), Cytochrome complex (Cyt-C), caspase-3 mRNA expression, cleaved-caspase-3 protein expression and apoptosis rate, as well as reducing B-cell lymphoma-2 (Bcl-2) mRNA expression and ratio of Bcl-2 to Bax, and triggered ferroptosis by upregulating Acyl-CoA Synthetase Long Chain Family Member 4 (ACSL4), transferrin receptor (TFR1) and Prostaglandin-Endoperoxide Synthase 2 (PTGS2) expression levels, and downregulating ferritin heavy chain 1 (FTH1), ferritin light chain 1 (FTL1), ferroportin 1 (FPN1), solute carrier family 7 member 11 (SCL7A11) and glutathione peroxidase 4 (GPX4) expression levels. The most obvious changes of these indexes were observed in co-treated group. Altogether, the results announced that Mo or Cd or both evoked apoptosis and ferroptosis by inhibiting Nrf2 pathway in the testis of ducks, and co-exposure to Mo and Cd exacerbated these variations.

Co-exposure to Environmentally Relevant Levels of Molybdenum and Cadmium Induces Oxidative Stress and Ferroptosis in the Ovary of Ducks.

Excessive doses of molybdenum (Mo) and cadmium (Cd) have toxic effects on animals. Nevertheless, the reproductive toxicity elicited by Mo and Cd co-exposure remains obscure. To evaluate the co-induce toxic impacts of Mo and Cd on ovaries, 8-day-old 40 healthy ducks were stochastically distributed to four groups and were raised a basal diet supplemented with Cd (4 mg/kg Cd) and/or Mo (100 mg/kg Mo). In the 16th week, ovary tissues were gathered. The data revealed that Mo and/or Cd decreased GSH content, CAT, T-SOD, and GSH-Px activities and increased MDA and H2O2 levels. Moreover, there was a significant decrease in nuclear Nrf2 protein level and its related downstream factors, while cytoplasmic Nrf2 protein level showed a substantial increase. Additionally, a marked elevation was observed in ferrous ion content and TFRC, GCLC, SLC7A11, ACSL4, and PTGS2 expression levels, while FTH1, FTL1, FPN1, and GPX4 expression levels were conversely reduced. These indicators exhibited more marked changes in the joint exposure group. In brief, our results announced that Mo and/or Cd resulted in oxidative stress and ferroptosis in duck ovaries. Synchronously, the Cd and Mo mixture intensified the impacts.

Role of transcription factor complex OsbHLH156-OsIRO2 in regulating manganese, copper, and zinc transporters in rice.

Iron (Fe), manganese (Mn), copper (Cu), and zinc (Zn) are essential micronutrients that are necessary for plant growth and development, but can be toxic at supra-optimal levels. Plants have evolved a complex homeostasis network that includes uptake, transport, and storage of these metals. It was shown that the transcription factor (TF) complex OsbHLH156-OsIRO2 is activated under Fe deficient conditions and acts as a central regulator on Strategy II Fe acquisition. In this study, the role of the TF complex on Mn, Cu, and Zn uptake was evaluated. While Fe deficiency led to significant increases in shoot Mn, Cu, and Zn concentrations, the increases of these divalent metal concentrations were significantly suppressed in osbhlh156 and osiro2 mutants, suggesting that the TF complex plays roles on Mn, Cu, and Zn uptake and transport. An RNA-sequencing assay showed that the genes associated with Mn, Cu, and Zn uptake and transport were significantly suppressed in the osbhlh156 and osiro2 mutants. Transcriptional activation assays demonstrated that the TF complex could directly bind to the promoters of OsIRT1, OsYSL15, OsNRAMP6, OsHMA2, OsCOPT1/7, and OsZIP5/9/10, and activate their expression. In addition, the TF complex is required to activate the expression of nicotianamine (NA) and 2'-deoxymugineic acid (DMA) synthesis genes, which in turn facilitate the uptake and transport of Mn, Cu, and Zn. Furthermore, OsbHLH156 and OsIRO2 promote Cu accumulation to partially restore the Fe-deficiency symptoms. Taken together, OsbHLH156 and OsIRO2 TF function as core regulators not only in Fe homeostasis, but also in Mn, Cu, and Zn accumulation.

HNRNPA2B1-mediated m6A modification of FOXM1 promotes drug resistance and inhibits ferroptosis in endometrial cancer via regulation of LCN2.

N6-methyladenosine (m6A) methylation is an extensive posttranscriptional RNA modification, and it is associated with various cellular responses, especially in tumor progression. An m6A "reader"-HNRNPA2B1 has been found oncogenic in multiple malignancies. As a key proliferation-related transcription factor, forkhead box protein M1 (FOXM1) is involved in tumorigenesis. Here, we elucidated the underlying mechanism by which HNRNPA2B1-mediated modification of FOXM1 promotes endometrial cancer (EC). The GSE115810 dataset was used to analyze the upregulated gene mRNA in late-stage EC tissues. The expression levels of HNRNPA2B1, FOXM1, and LCN2 in EC samples were shown by western blotting and qPCR. The interaction among HNRNPA2B1, FOXM1, and LCN2 in EC cells was detected using bioinformatics analysis, RNA immunoprecipitation (RIP), RNA pull-down, RNA decay analysis, and luciferase reporter experiments. Cisplatin (DDP)-resistant EC cells were constructed using HEC-1-A and HEC-1-B cells, named HEC-1-A/DDP and HEC-1-B/DDP, respectively. Proliferation, migration, and invasiveness in treated HEC-1-A/DDP and HEC-1-B/DDP cells were detected by EdU, wound healing, and transwell assays. Ferroptosis-resistant gene expression, MDA level, and ROS level were measured. The m6A modification level in EC tissues was elevated. HNRNPA2B1 and FOXM1 levels were upregulated in EC. HNRNPA2B1 expression was positively related to FOXM1 expression in EC samples, and HNRNPA2B1 bound to the 3'UTR of FOXM1 and stabilized FOXM1 mRNA via m6A modification. FOXM1 positively regulated LCN2 expression in EC cells by binding to the LCN2 promotor. Knockdown of FOXM1 downregulated ferroptosis-resistant gene expression and increased MDA and ROS levels in DDP-resistant EC cells. Rescue assays revealed that LCN2 overexpression eliminated the effects mediated by FOXM1 knockdown on the proliferation, migration, invasiveness, and ferroptosis in DDP-resistant EC cells. In conclusion, HNRNPA2B1-mediated mA modification of FOXM1 facilitates drug resistance and inhibits ferroptosis in EC cells by upregulating LCN2 expression.

Co-exposure to molybdenum and cadmium evokes necroptosis and decreases apoptosis in duck myocardium.

Superfluous molybdenum (Mo) and cadmium (Cd) in the environment are detrimental to organisms through their accumulation. The NF-κB/TNF-α axis plays a vital part in regulating necroptosis and apoptosis. However, the impacts of Mo and/or Cd on myocardium injury in ducks and the function of NF-κB/TNF-α axis are not clear in the process. In this research, ducks exposed to different dosages of Mo and/or Cd were applied as the study object. The findings substantiated that the accumulation of Mo and/or Cd caused elements imbalance and necroptosis in myocardial tissue. As p-NF-κB/TNF-α expression up-regulated, RIPK1/RIPK3/p-MLKL expression significantly increased in all treatment groups, while the expression of c-caspase-8/3 markedly decreased. Moreover, apoptosis rate obviously decreased in Cd treated groups and clearly elevated in Mo group. Mitochondria-mediated apoptosis was activated by excessive Mo and inhibited by Mo + Cd, but Cd exposure alone had little effect on it. Collectively, our research confirmed that Mo and/or Cd evoked necroptosis via NF-κB/TNF-α axis, and decreased death receptor-mediated apoptosis in duck myocardium, the impacts of Mo and/or Cd on mitochondrial-mediated apoptosis were different. These results are significant for studying toxicology of Mo and/or Cd and preserving the ecosystem.

MEN1 is a regulator of alternative splicing and prevents R-loop-induced genome instability through suppression of RNA polymerase II elongation.

The fidelity of alternative splicing (AS) patterns is essential for growth development and cell fate determination. However, the scope of the molecular switches that regulate AS remains largely unexplored. Here we show that MEN1 is a previously unknown splicing regulatory factor. MEN1 deletion resulted in reprogramming of AS patterns in mouse lung tissue and human lung cancer cells, suggesting that MEN1 has a general function in regulating alternative precursor mRNA splicing. MEN1 altered exon skipping and the abundance of mRNA splicing isoforms of certain genes with suboptimal splice sites. Chromatin immunoprecipitation and chromosome walking assays revealed that MEN1 favored the accumulation of RNA polymerase II (Pol II) in regions encoding variant exons. Our data suggest that MEN1 regulates AS by slowing the Pol II elongation rate and that defects in these processes trigger R-loop formation, DNA damage accumulation and genome instability. Furthermore, we identified 28 MEN1-regulated exon-skipping events in lung cancer cells that were closely correlated with survival in patients with lung adenocarcinoma, and MEN1 deficiency sensitized lung cancer cells to splicing inhibitors. Collectively, these findings led to the identification of a novel biological role for menin in maintaining AS homeostasis and link this role to the regulation of cancer cell behavior.

Exposure of zebrafish (Danio rerio) to trans-2-hexenal induces oxidative stress and protein degeneration of the gill.

Trans-2-hexenal (T2H) has great commercial value for development as a biopesticide, but its toxicity risk to nontarget organisms is unknown. Here, the toxicity and underlying mechanism of T2H on zebrafish (Danio rerio) were investigated. The LC50 (48 h) of T2H on zebrafish is 4.316 µg/mL, and the aldehyde group is essential to its toxicity. In 14-day chronic toxicity tests, 0.432 µg/mL T2H resulted in a higher mortality of zebrafish than the control group. Furthermore, the sensitivity of zebrafish to different administration methods was gill administration>oral administration>transdermal administration>intravenous injection. T2H induced significant cell death and ROS generation in zebrafish gill cells in a concentration-dependent manner. After treatment with 4.316 µg/mL T2H, the expression of oxidative stress-related genes (nrf2, gstp1, keap1b, sod1 and sod2) and the content of malondialdehyde (MDA) were up-regulated. Incubation with T2H caused an immediate denaturation of gill protein, which was aggravated with increasing dose of T2H. We also found that T2H at 21.225 mg/mL significantly reduced the in vitro activity of succinate dehydrogenase (SDH). Among the three amino acids tested, T2H was only found to react with methionine and glycine to form adducts, which may be the basis of the protein denaturation. This study confirmed that T2H could induce oxidative stress and protein denaturation in zebrafish gills, providing important information for risk assessment of T2H exposure.

SDH mutations confer complex cross-resistance patterns to SDHIs in Corynespora cassiicola.

Succinate dehydrogenase inhibitors (SDHIs) are one of the most frequently used fungicides in cucumber fields in China. Our previous studies indicated that the sensitivity profile of Corynespora cassiicola, the causal agent of Corynespora leaf spot, to different SDHIs varied greatly; however, the underlying mechanism remains unclear. The 50% effective concentration (EC50) values of boscalid, fluopyram, fluxapyroxad and isopyrazam in C. cassiicola collected from 2017 to 2020 shifted, with resistance frequencies of 79.83%, 78.43%, 83.19% and 49.86%, respectively. The sequence alignment of sdhB/C/D of resistant strains revealed that eight single amino acid mutations (B-H278Y/L, B-I280V, C-S73P, C-N75S, C-H134R, D-D95E and D-G109V), and three dual-mutations (B-I280V&C-S73P, B-I280V&C-N75S and C-S73P&C-N75S) conferred various SDHI resistance levels and cross-resistance profiles. The expression level of the sdhB/C/D gene and succinate dehydrogenase (SDH) activity in the mutants were significantly altered by the presence of SDHIs, compared with the wild type strain. Additionally, molecular docking results suggested that the missense mutation influenced the crystal structure of SDH and subsequently interfered with the interaction bonds and bond distances among the target protein and chemicals. In brief, amino acid mutations altered the fungicide response of target gene expression, SDH activity and the binding features of SDH-ligand complexes and subsequently conferred multiple resistance levels and complex cross-resistance patterns to SDHIs in C. cassiicola. The evaluation of C. cassiicola resistance to SDHIs provided a significant foundation for efficient chemical development and integrated CLS management strategies.

Loss of MEN1 leads to renal fibrosis and decreases HGF-Adamts5 pathway activity via an epigenetic mechanism.

BACKGROUND: Renal fibrosis is a serious condition that results in the development of chronic kidney diseases. The MEN1 gene is an epigenetic regulator that encodes the menin protein and its role in kidney tissue remains unclear. METHODS: Kidney histology was examined on paraffin sections stained with hematoxylin-eosin staining. Masson's trichrome staining and Sirius red staining were used to analyze renal fibrosis. Gene and protein expression were determined by quantitative real-time PCR (qPCR) and Western blot, respectively. Immunohistochemistry staining in the kidney tissues from mice or patients was used to evaluate protein levels. Flow cytometry was used to analyze the cell cycle distributions and apoptosis. RNA-sequencing was performed for differential expression genes in the kidney tissues of the Men1f/f and Men1∆/∆ mice. Chromatin immunoprecipitation sequencing (ChIP-seq) was carried out for identification of menin- and H3K4me3-enriched regions within the whole genome in the mouse kidney tissue. ChIP-qPCR assays were performed for occupancy of menin and H3K4me3 at the gene promoter regions. Luciferase reporter assay was used to detect the promoter activity. The exacerbated unilateral ureteral obstruction (UUO) models in the Men1f/f and Men1∆/∆ mice were used to assess the pharmacological effects of rh-HGF on renal fibrosis. RESULTS: The expression of MEN1 is reduce in kidney tissues of fibrotic mouse and human diabetic patients and treatment with fibrotic factor results in the downregulation of MEN1 expression in renal tubular epithelial cells (RTECs). Disruption of MEN1 in RTECs leads to high expression of α-SMA and Collagen 1, whereas MEN1 overexpression restrains epithelial-to-mesenchymal transition (EMT) induced by TGF-ß treatment. Conditional knockout of MEN1 resulted in chronic renal fibrosis and UUO-induced tubulointerstitial fibrosis (TIF), which is associated with an increased induction of EMT, G2/M arrest and JNK signaling. Mechanistically, menin recruits and increases H3K4me3 at the promoter regions of hepatocyte growth factor (HGF) and a disintegrin and metalloproteinase with thrombospondin motifs 5 (Adamts5) genes and enhances their transcriptional activation. In the UUO mice model, exogenous HGF restored the expression of Adamts5 and ameliorated renal fibrosis induced by Men1 deficiency. CONCLUSIONS: These findings demonstrate that MEN1 is an essential antifibrotic factor in renal fibrogenesis and could be a potential target for antifibrotic therapy.

Green Leaf Volatile Trans-2-Hexenal Inhibits the Growth of Fusarium graminearum by Inducing Membrane Damage, ROS Accumulation, and Cell Dysfunction.

Fusarium graminearum, the main agent of Fusarium head blight (FHB), can cause serious yield loss and secrete mycotoxins to contaminate grain. Here, the biological activity of trans-2-hexenal (T2H) against F. graminearum was determined and its mode of action (MOA) was investigated. Furthermore, surface plasmon resonance with liquid chromatography-tandem mass spectrometry (SPR-LC-MS/MS), bioinformatic analysis, and gene knockout technique were combined to identify the binding proteins of T2H in F. graminearum cells. T2H exhibited satisfactory inhibitory activity against F. graminearum in vitro. Good lipophilicity greatly enhanced the affinity of T2H to F. graminearum mycelia and further caused membrane damage. The FgTRR (thioredoxin reductase) gene negatively regulates the sensitivity of F. graminearum to T2H by reducing the generation of reactive oxygen species (ROS) induced by T2H. Two mutant strains with FgSLX1 (structure-specific endonuclease subunit) and FgCOPB (coatomer subunit ß) genes knockout showed decreased sensitivity to T2H, suggesting that these two genes may be involved in the antimicrobial activity of T2H. Taken together, T2H can inhibit F. graminearum growth by multiple MOAs and can be used as a biofumigant to control the occurrence of FHB in the field.

Phosphonium ionic liquid-promoted Ce-doped nitrogen-rich TiO2for degradation gaseous pollutant under visible light irradiation.

Altering physicochemical properties of TiO2based on modifying the cation and anion structure of ionic liquids (ILs) is of great interests for environment. Up to date, the research involving IL-assisted synthesis of TiO2was focused on imidazolium IL, and much less attention was devoted to IL with other structures. Hence, strategy for preparation of TiO2in phosphonium IL is presented to control the growth of TiO2nanocrystals. The as-prepared noble cerium-doped nitrogen-rich phosphonium IL-TiO2photocatalyst with assisted by tributyl(propyl)phosphonium tetrafluoroborate exhibits a higher specific surface area and smaller crystallite size, which is conducive to the production of more and faster active substance, such as hydroxyl oxygen. When evaluated for photocatalysis of gaseous toluene under visible light irradiation, the sample manifests high degradation rate and efficiency, as well as excellent recycling performance due to the existence of superoxide radical produced by the Ce3+/Ce4+redox reaction. The introduction of phosphonium IL and Ce greatly enhanced charge separation efficiency and promoted production of active substances. Nitrogen also existed in the form of interstitial nitrogen and substituted nitrogen improves its response to visible light. This work shows promising application of phosphonium IL for highly enhanced TiO2photocatalytic performance.

Ecotoxicological effects of pyraclostrobin on tilapia (Oreochromis niloticus) via various exposure routes.

Pyraclostrobin is a widely used and highly efficient fungicide that also has high toxicity to aquatic organisms, especially fish. Although some research has reported the toxic effects of pyraclostrobin on fish, the main toxic pathways of pyraclostrobin in fish remain unclear. The present study has integrated histopathological, biochemical and hematological techniques to reveal the main toxic pathways and mechanisms of pyraclostrobin under different exposure routes. Our results indicated that pyraclostrobin entered fish mainly through the gills. The highest accumulation of pyraclostrobin was observed in the gills and heart compared with accumulation in other tissues and gill tissue showed the most severe damage. Hypoxia symptoms (water jacking, tummy turning and cartwheel formation) in fish were observed throughout the experiment. Taken together, our results suggested that the gills are important target organs. The high pyraclostrobin toxicity to gills might be associated with oxidative damage to the gills, inducing alterations in ventilation frequency, oxygen-carrying substances in blood and disorders of energy metabolism. Our research facilitates a better understanding of the toxic mechanisms of pyraclostrobin in fish, which can promote the ecotoxicological research of agrochemicals on aquatic organisms.

DNA methylation is involved in acclimation to iron-deficiency in rice (Oryza sativa).

Iron (Fe) is an essential micronutrient in plants, and Fe limitation significantly affects plant growth, yield and food quality. While many studies have reported the transcriptomic profile and pursue molecular mechanism in response to Fe limitation, little is known if epigenetic factors play a role in response to Fe-deficiency. In this study, whole-genome bisulfite sequencing analysis, high-throughput RNA-Seq of mRNA, small RNA and transposable element (TE) expression with root and shoot organs of rice seedlings under Fe-sufficient and Fe-deficient conditions were performed. The results showed that widespread hypermethylation, especially for the CHH context, occurred after Fe-deficiency. Integrative analysis of methylation and transcriptome revealed that the transcript abundance of Fe-deficiency-induced genes was negatively correlated with nearby TEs and positively with the 24-nucleotide siRNAs. The ability of methylation to affect the physiology and molecular response to Fe-deficiency was tested using an exogenous DNA methyltransferase inhibitor (5-azacytidine), and genetically using a mutant for domains rearranged methyltransferase 2 (DRM2), that lacks CHH methylation. Both approaches resulted in decreased growth and Fe content in rice plants. Thus, alterations in specific methylation patterns, directed by siRNAs, play an important role in acclimation of rice to Fe-deficient conditions. Furthermore, comparison with other reports suggests this may be a universal mechanism to acclimate to limited nutrient availability.

Evaluation of the antifungal and biochemical activities of mefentrifluconazole against Botrytis cinerea.

Mefentrifluconazole is the first product of a new sub-class of triazoles fungicides, i.e., the isopropanol triazoles, with the broad spectrum and high activity. In this study, the potential and biochemical activities of mefentrifluconazole against Botrytis cinerea were investigated. The frequency distribution of all EC50 values of mefentrifluconazole against mycelial growth and germ tube elongation of 106 isolates formed unimodal curve, with the mean EC50 values of 0.124 ± 0.025 and 0.015 ± 0.008 µg/mL, respectively. The effect of mefentrifluconazole against gray mold was determined on detached leaves of cucumber in vivo, the treatment of mefentrifluconazole at 200 µg/mL provided 100% preventative efficacy and 72.7% curative efficacy. No evident correlation was detected between the sensitivity of B. cinerea to mefentrifluconazole and that to tebuconazole, difenoconazole, myclobutanil, hexaconazole, triadimefon, flusilazole and pyrisoxazole (P > 0.05). Mefentrifluconazole treatment resulted in the increase of mycelium branch, the decrease of ergosterol content and the changes of the permeability of cell membrane. These studies evaluated the potential of mefentrifluconazole to control gray mold and helped us to understand the possible biochemical activity of mefentrifluconazole against B.cinerea.

The Bioactivity and Efficacy of Benzovindiflupyr Against Corynespora cassiicola, the Causal Agent of Cucumber Corynespora Leaf Spot.

Corynespora cassiicola, which causes Corynespora leaf spot, results in considerable yield loss of cucumber grown in greenhouses. Frequent reports of reduced efficacy and control failure of fungicides warrant new, efficient alternative chemistries. In this study, the sensitivity of C. cassiicola to benzovindiflupyr was evaluated using a collection of 81 isolates collected from Shandong, China. The mean EC50 values for mycelial growth, spore germination, and germ tube elongation of C. cassiicola were 0.69 ± 0.44, 0.12 ± 0.063, and 0.13 ± 0.076 µg ml-1, respectively. Benzovindiflupyr treatment led to a reduced respiration rate and ATP production of C. cassiicola and decreased spore pathogenicity by 21.9% on average. Additionally, detached cucumber leaves sprayed with fungicides before or after inoculation were used to assess the efficacy of benzovindiflupyr against C. cassiicola. Benzovindiflupyr (150 µg ml-1) exhibited preventive and curative efficacies of 86.9 and 77.1%, respectively. Benzovindiflupyr at 150 g a.i. ha-1 provided over 70% efficacy in field trials performed in 2018 and 2019, which was significantly higher than that of the reference fungicides fluopyram and fluxapyroxad at the same dose. Furthermore, the yield of commercial cucumber increased as disease incidence decreased. Our findings pave the way for the introduction of benzovindiflupyr in the integrated management of Corynespora leaf spot.

Toxicity, residue and risk assessment of tetraniliprole in soil-earthworm microcosms.

Maize seed treatment with chemicals to control underground pests is a common agricultural practice, but inappropriate use of insecticides poses a considerable threat to plant development and soil nontarget organisms. In this study, the availability of tetraniliprole seed dressing to control the black cutworm Agrotis ipsilon (Lepidoptera: Noctuidae) in the maize seeding stage and its safety to earthworms (Eisenia fetida) were investigated. The selective toxicity (ST) of tetraniliprole between E. fetida and A. ipsilon was greater than 4000. No significant adverse effect of tetraniliprole seed treatment on the germination of maize seeds was observed at concentrations of 2.4-9.6 g a.i. /kg seed. Compared with the untreated control, seed treatment with tetraniliprole at 9.6 g a.i. /kg seed greatly reduced the percentage of damaged plants from 88.73% to 26.67%, and achieved the highest control effect of 69.91%. Tetraniliprole of 2.4 g a.i. /kg seed can effectively inhibit A. ipsilon until 14 days after seed germination, with the lowest mortality rate of 44.44%. During the entire exposure period, the maximum residual concentration of tetraniliprole detected in the soil (5.86 mg/kg) was considerably lower than the LC50 value of tetraniliprole to E. fetida (>4000 mg/kg). According to the low-tier risk assessment, the highest risk quotient (RQ) of tetraniliprole seed treatment to earthworms at test concentrations was 2.8 × 10-3, which was evaluated as acceptable. This study provided data support for tetraniliprole seed treatment to control underground pests in maize fields.

Activity of the Novel Fungicide Mefentrifluconazole Against Colletotrichum scovillei.

The prevalence and destructiveness of anthracnose, caused by Colletotrichum scovillei, in pepper production regions seriously affects pepper yield and quality. Mefentrifluconazole, the first of the isopropanol-azole subgroup of triazole fungicides, was introduced for the control of pepper anthracnose. However, the growth characteristics of pepper fruit and rapid spread of anthracnose suggest that the fungicide application method must be optimized to enhance fungicide efficacy. The sensitivity of C. scovillei to mefentrifluconazole was determined by mycelial growth and germ tube elongation assays using 157 single-spore isolates with mean 50% effective concentration values of 0.462 ± 0.138 and 0.359 ± 0.263 mg/liter, respectively. The in vivo data also showed that mefentrifluconazole had favorable protective and curative effects against pepper anthracnose. Mefentrifluconazole significantly affected C. scovillei infection on pepper by reducing appressorium formation and sporulation, shriveling spores and germ tubes, and causing the abnormal development of appressoria and conidiophores. Mefentrifluconazole could move acropetally, horizontally, and basipetally in pepper plants. Compared with a knapsack sprayer, mefentrifluconazole applied by mist sprayer exhibited significantly better activity against pepper anthracnose. Additionally, as the spray volume increased from 45 to 150 liters/ha, the control efficacy of mefentrifluconazole first increased and then tended to be steady, with an optimal spray volume of 90 liters/ha. The difference in disease control efficacy was related to the deposition and droplet distribution of mefentrifluconazole on the pepper fruit. These results provide scientific guidance for the application of mefentrifluconazole in pepper fields and improved fungicide utilization.

Evaluation of Sensitivity and Resistance Risk of Corynespora cassiicola to Isopyrazam and Mefentrifluconazole.

Necrotic lesions on leaves caused by Corynespora leaf spot (CLS) seriously threaten the quality and yield of cucumber in China. Corynespora cassiicola has developed different degrees of sensitivity to various fungicides due to its long-term and extensive application. In our work, the effect of isopyrazam and mefentrifluconazole on different life stages of C. cassiicola was examined. To determine the optimal effect of binary mixtures of isopyrazam and mefentrifluconazole, the two fungicides were mixed at different proportions. Furthermore, the disease suppression of isopyrazam, mefentrifluconazole, and their compound mixture against CLS was evaluated in greenhouse experiments. Ultraviolet (UV) mutagenesis and fungicide-selection methods were performed to assess the risk of resistance development. Among the three life stages tested, isopyrazam showed the weakest inhibition on mycelial growth, and mefentrifluconazole showed the strongest inhibition of germ tube elongation. According to Wadley's and cotoxicity coefficient methods, the optimal proportion of the two-component mixture of isopyrazam and mefentrifluconazole was 1:1. Isopyrazam, mefentrifluconazole, and their binary mixture at 1:1 reduced the disease severity of CLS on potted cucumber plants, with protective effects of 31.11, 24.65, and 42.12% and curative effect of 33.90, 37.48, and 42.84%, respectively. Compared with isopyrazam or mefentrifluconazole alone, the binary mixture of the two fungicides at 1:1 did not exert significant influence on the change of C. cassiicola sensitivity. Undoubtedly, such data will greatly facilitate the screening of new fungicides for CLS and resistance management.

The relationship between features enabling SDHI fungicide binding to the Sc-Sdh complex and its inhibitory activity against Sclerotinia sclerotiorum.

BACKGROUND: A new generation of succinate dehydrogenase inhibitors (SDHIs) with high efficiency and broad-spectrum antifungal activity has been frequently used in crop production. Sclerotinia stem rot is a major disease of various plants and crops caused by Sclerotinia sclerotiorum. Although benzovindiflupyr and isopyrazam reportedly have high activity against S. sclerotiorum, little is known about the bioactivity of different SDHIs classes against S. sclerotiorum or the mechanism of their differential antifungal activity. RESULTS: The in vitro tests revealed that the pyrazole-4-carboxamides of SDHIs (benzovindiflupyr, isopyrazam, fluxapyroxad, pydiflumetofen) had the highest activity against S. sclerotiorum followed by pyridine carboxamides (boscalid), pyridinyl-ethyl benzamides (fluopyram) and thiazole carboxamides (thifluzamide), and of these thifluzamide showed poor antifungal activity with EC50 values greater than 6.01 mg L-1 . The pyrazole-4-carboxamides of SDHIs showed satisfactory protective and curative activity against Sclerotinia stem rot. After treatment with the pyrazole-4-carboxamides of SDHIs, mitochondrial function in S. sclerotiorum decreased significantly. The enzyme activity assays revealed a lower affinity between thifluzamide and the Sc-Sdh complex than was observed for the other six fungicides, with IC50 values ranging from 0.0036 to 1.2088 µmol L-1 . Additionally, the docking positions of fungicides were similar, yet binding energies were different in the docking study with the Sdh complex. The correspondingly weaker hydrogen bonds may be responsible for the poor activity of thifluzamide against S. sclerotiorum. CONCLUSION: Understanding different binding features of various SDHIs classes with the Sc-Sdh complex might be beneficial for the design and development of highly effective broad-spectrum fungicides to ensure high yield and quality in crops by reducing fungicide use. © 2020 Society of Chemical Industry.

Development of a LAMP method for detecting the N75S mutant in SDHI-resistant Corynespora cassiicola.

The replacement of asparagine with serine at codon 75 of the sdhC gene (N75S) confers succinate dehydrogenase inhibitor resistance in Corynespora cassiicola, which caused by consecutive fungicide application. To rapidly detect the mutation of N75S, a method based on loop-mediated isothermal amplification (LAMP) was developed in this study. The optimal primer set among the six primer sets designed could clearly identify N75S from the wild-type genotype. The detection threshold of the optimized LAMP mixture (10 µL) was 8.8 fg of target DNA at 63 °C within 60 min. This method specifically showed a color change and ladder-like band only when DNA extracted from isolates containing the N75S mutation was added. The results of stability tests suggested a satisfactory repeatability of this method. Additionally, the assay could positively distinguish N75S mutants from crude DNA isolated from conidia and mycelia of C. cassiicola. Given the high efficiency, sensitivity, specificity, repeatability and simplicity of operation, the LAMP method established here could be useful to evaluate the shift in the sensitivity of C. cassiicola to SDHIs and will provide significant data for the management of Corynespora leaf spot.