Prevalence of Azole-Resistant Aspergillus fumigatus is Highly Associated with Azole Fungicide Residues in the Fields.

Triazole resistance in Aspergillus fumigatus is a growing public health concern. In addition to its emergence in the therapy of invasive aspergillosis by triazole medicines, it has been frequently detected in agricultural fields all over the world. Here, we explore the potential link between residues of azole fungicides with similar chemical structure to triazole medicines in soil and the emergence of resistant A. fumigatus (RAF) through 855 500 km2 monitoring survey in Eastern China covering 6 provinces. In total, 67.3%, 15.2%, 12.3%, 2.9%, 1.5%, 0.4%, and 0.3% of the soil samples contained these five fungicides (tebuconazole, difenoconazole, propiconazole, hexaconazole, and prochloraz) of 0-100, 100-200, 200-400, 400-600, 600-800, 800-1000, and >1000 ng/g, respectively. The fractions of samples containing RAF isolates were 2.4%, 5.2%, 6.4%, 7.7%, 7.4%, 14.3%, and 20.0% of the samples with total azole fungicide residues of 0-100, 100-200, 200-400, 400-600, 600-800, 800-1000, and >1000 ng/g, respectively. We find that the prevalence of RAFs is positively (P < 0.0001) correlated with residual levels of azole fungicides in soils. Our results suggest that the use of azole fungicides in agriculture should be minimized and the intervals between treatments expanded to reduce the selective pressure toward the development of resistance in A. fumigatus in agricultural fields.

Endophytic fungus Falciphora oryzae promotes lateral root growth by producing indole derivatives after sensing plant signals.

The endophytic fungus Falciphora oryzae was initially isolated from wild rice (Oryza granulata) and colonizes many crop species and promotes plant growth. However, the molecular mechanisms underlying F. oryzae-mediated growth promotion are still unknown. We found that F. oryzae was able to colonize Arabidopsis thaliana. The most dramatic change after F. oryzae inoculation was observed in the root architecture, as evidenced by increased lateral root growth but reduced primary root length, similar to the effect of auxin, a significant plant growth hormone. The expression of genes responsible for auxin biosynthesis, transport, and signalling was regulated in Arabidopsis roots after F. oryzae cocultivation. Indole derivatives were detected at significantly higher levels in liquid media after cocultivation compared with separate cultivation of Arabidopsis and F. oryzae. Consistently, the expression of indole biosynthetic genes was highly upregulated in F. oryzae upon treatment with Arabidopsis exudates. Global analysis of Arabidopsis gene expression at the early stage after F. oryzae cocultivation suggested that signals were exchanged to initiate Arabidopsis-F. oryzae interactions. All these results suggest that signalling molecules from Arabidopsis roots are perceived by F. oryzae and induce the biosynthesis of indole derivatives in F. oryzae, consequently stimulating Arabidopsis lateral root growth.

Vision-Based Moving Obstacle Detection and Tracking in Paddy Field Using Improved Yolov3 and Deep SORT.

Using intelligent agricultural machines in paddy fields has received great attention. An obstacle avoidance system is required with the development of agricultural machines. In order to make the machines more intelligent, detecting and tracking obstacles, especially the moving obstacles in paddy fields, is the basis of obstacle avoidance. To achieve this goal, a red, green and blue (RGB) camera and a computer were used to build a machine vision system, mounted on a transplanter. A method that combined the improved You Only Look Once version 3 (Yolov3) and deep Simple Online and Realtime Tracking (deep SORT) was used to detect and track typical moving obstacles, and figure out the center point positions of the obstacles in paddy fields. The improved Yolov3 has 23 residual blocks and upsamples only once, and has new loss calculation functions. Results showed that the improved Yolov3 obtained mean intersection over union (mIoU) score of 0.779 and was 27.3% faster in processing speed than standard Yolov3 on a self-created test dataset of moving obstacles (human and water buffalo) in paddy fields. An acceptable performance for detecting and tracking could be obtained in a real paddy field test with an average processing speed of 5-7 frames per second (FPS), which satisfies actual work demands. In future research, the proposed system could support the intelligent agriculture machines more flexible in autonomous navigation.

Microenvironmental Interplay Predominated by Beneficial Aspergillus Abates Fungal Pathogen Incidence in Paddy Environment.

Rice fungal pathogens, responsible for severe rice yield loss and biotoxin contamination, cause increasing concerns on environmental safety and public health. In the paddy environment, we observed that the asymptomatic rice phyllosphere microenvironment was dominated by an indigenous fungus, Aspergillus cvjetkovicii, which positively correlated with alleviated incidence of Magnaporthe oryzae, one of the most aggressive plant pathogens. Through the comparative metabolic profiling for the rice phyllosphere microenvironment, two metabolites were assigned as exclusively enriched metabolic markers in the asymptomatic phyllosphere and increased remarkably in a population-dependent manner with A. cvjetkovicii. These two metabolites evidenced to be produced by A. cvjetkovicii in either a phyllosphere microenvironment or artificial media were purified and identified as 2(3H)-benzofuranone and azulene, respectively, by gas chromatography coupled to triple quadrupole mass spectrometry and nuclear magnetic resonance analyses. Combining with bioassay analysis in vivo and in vitro, we found that 2(3H)-benzofuranone and azulene exerted dissimilar actions at the stage of infection-related development of M. oryzae. A. cvjetkovicii produced 2(3H)-benzofuranone at the early stage to suppress MoPer1 gene expression, leading to inhibited mycelial growth, while azulene produced lately was involved in blocking of appressorium formation by downregulation of MgRac1. More profoundly, the microenvironmental interplay dominated by A. cvjetkovicii significantly blocked M. oryzae epidemics in the paddy environment from 54.7 to 68.5% (p < 0.05). Our study first demonstrated implication of the microenvironmental interplay dominated by indigenous and beneficial fungus to ecological balance and safety of the paddy environment.

Biotoxin Tropolone Contamination Associated with Nationwide Occurrence of Pathogen Burkholderia plantarii in Agricultural Environments in China.

Tropolone, a biotoxin produced by the agricultural pathogen Burkholderia plantarii, exerts cytotoxicity toward a wide array of biota. However, due to the lack of quantitative and qualitative approach, both B. plantarii occurrence and tropolone contamination in agricultural environments remain poorly understood. Here, we presented a sensitive and reliable method for detection of B. plantarii in artificial, plant, and environmental matrices by tropolone-targeted gas chromatography-triple-quadrupole tandem mass spectrometry analysis. Limits of detection for B. plantarii and tropolone were 10 colony-forming units (CFU)/mL and 0.017 µg/kg, respectively. In a series of simulation trials, we found that B. plantarii from 10 to 108 CFU/mL produced tropolone between 0.006 and 107.8 mg/kg in a cell-population-dependent manner, regardless of habitat. Correlation analysis clarified a reliable reflection of B. plantarii density by tropolone level with R2 values from 0.9201 to 0.9756 ( p < 0.01). Through a nationwide pilot study conducted in China, tropolone contamination was observed at 0.014-0.157 mg/kg in paddy soil and rice grains, and subsequent redundancy analysis revealed soil organic matter to be a dominant environmental factor, having a positive correlation with tropolone contamination. In this context, our results imply that potential ecological and dietary risks posed by long-term exposure to trace levels of tropolone contamination are of concern.

Biochemical and histological alterations in adult zebrafish（Danio rerio）ovary following exposure to the tetronic acid insecticide spirotetramat.

As a new tetronic acid derivative insecticide, spirotetramat has been reported to be toxic to an array of aquatic organisms. However, the toxic effects of spirotetramat on zebrafish especially at ovary are still obscure. Hereby, the acute toxicity of spirotetramat towards zebrafish（Danio rerio），as well as the changes on biochemical and histological traits of ovary were investigated. The acute toxicity test results showed that the median lethal concentration (LC50) value of spirotetramat were 9.61 mg/L and 7.21 mg/L at 72 h and 96 h, respectively, suggesting spirotetramat has moderate toxicity to zebrafish. In the following sub-lethal toxicity test, the gene expression of superoxide dismutase (SOD), catalase (CAT), and gonadotropic hormone receptor (FSHR and LHR) together with the content of malondialdehyde (MDA) in ovary were measured at 14, 21, and 28 days after exposure to 36, 360 and 720 µg/L. Under high concentration treatment (360 and 720 µg/L), MDA content, the relative transcription CAT and SOD gene level increased significantly in ovary (p < 0.05). That indicated sub-lethal doses spirotetramat caused oxidative stress and lipid peroxidation in zebrafish ovary during the entire experimental period. Under the exposure to spirotetramat at 720 µg/L after 14 days, the relative transcript FSHR gene level was down regulated, and the relative transcript LHR gene level was up regulated. Moreover, spirotetramat affected the oocyte development especially on the diameter size and maturation during the ovary tissue biopsies at 28 days. Taken together, these findings revealed the adverse effects of spirotetramat on fish from the biochemical and histological aspects.

Residual behavior and risk assessment of the mixed formulation of benzene kresoxim-methyl and fluazinam in cucumber field application.

Benzene kresoxim-methyl (BKM) is a new strobilurin fungicide mixed with fluazinam (Flu) into 40 % suspension concentrate (SC) formulation to improve fungicidal efficacy and to reduce the risk of resistance on cucumber. However, the fate of the fungicide residues in a cucumber plantation remains unclear. Thus, an efficient method of ultra-performance liquid chromatography combined with a modified quick, easy, cheap, effective, rugged, and safe sample preparation was developed to simultaneously determine the BKM and Flu residues in cucumber and soil samples to investigate their residual behavior and risk assessment in the cucumber plantation. This analytical method revealed that the detection limits of BKM and Flu were 1.64 × 10(-3) and 1.82 × 10(-3) mg L(-1), respectively, and their average recoveries in the cucumber and soil samples were 77.5-106.9 %. The respective half-lives of BKM and Flu were 2.2-3.4 and 1.0-2.5 days in cucumber; in soil, the half-lives of BKM and Flu were 2.6-5.0 and 2.4-5.3 days, respectively. Seven days after application, the terminal residues of BKM and Flu in cucumber were less than 0.02 mg kg(-1). The residual profiles of BKM and Flu suggested that these fungicides could rapidly degrade in cucumber plantations. Their hazard quotient values were all less than 1 on the preharvest intervals of 3, 5, and 7 days, indicating that the dietary risk of BKM and Flu 40 % SC with the recommended usage on cucumber is negligible.

Nitrous oxide emission potentials of Burkholderia species isolated from the leaves of a boreal peat moss Sphagnum fuscum.

Using a culture-based nitrous oxide (N2O) emission assay, three active N2O emitters were isolated from Sphagnum fuscum leaves and all identified as members of Burkholderia. These isolates showed N2O emission in the medium supplemented with [Formula: see text] but not with [Formula: see text], and Burkholderia sp. SF-E2 showed the most efficient N2O emission (0.20 µg·vial(-1)·day(-1)) at 1.0 mM KNO3. In Burkholderia sp. SF-E2, the optimum pH for N2O production was 5.0, close to that of the phyllosphere of Sphagnum mosses, while the optimum temperature was uniquely over 30 °C. The stimulating effect of additional 1.5 mM sucrose on N2O emission was ignorable, but Burkholderia sp. SF-E2 upon exposure to 100 mg·L(-1) E-caffeic acid showed uniquely 67-fold higher N2O emission. All of the three N2O emitters were negative in both acetylene inhibition assay and PCR assay for nosZ-detection, suggesting that N2O reductase or the gene itself is missing in the N2O-emitting Burkholderia.

Mitochondrial dynamics disruption: Unraveling Dinotefuran's impact on cardiotoxicity.

Exposure to pesticides has been associated with several cardiovascular complications in animal models. Neonicotinoids are now the most widely used insecticide globally, while the impact of neonicotinoids on cardiovascular function and the role of mitochondrial dynamics in neonicotinoids-induced cardiotoxicity is unclear. In the present study, Xenopus laevis tadpoles were exposed to environmental related concentrations (0, 5, and 50 µg/L) of typical neonicotinoid dinotefuran, with two enantiomers, for 21 days. We evaluated the changes in heart rate and cardiomyocyte apoptosis in exposed tadpoles. Then, we performed the transcriptome, metabolomics, transmission electron microscopy (TEM), and protein immunoblot to investigate the potential adverse impact of two enantiomers of dinotefuran on cardiotoxicity associated with mitochondrial dynamics. We observed changes in heart rate and increased cardiomyocyte apoptosis in exposed tadpoles, indicating that dinotefuran had a cardiotoxic effect. We further found that the cardiac contractile function pathway was significantly enriched, while the glucose metabolism-related pathways were also disturbed significantly. TEM observation revealed that the mitochondrial morphology of cardiomyocytes in exposed tadpoles was swollen, and mitophagy was increased. Mitochondria fusion was excessively manifested in the enhanced mitochondrial fusion protein. The mitochondrial respiratory chain was also disturbed, which led to an increase in ROS production and a decrease in ATP content. Therefore, our results suggested that dinotefuran exposure can induce cardiac disease associated mitochondrial disorders by interfering with the functionality and dynamics of mitochondria. In addition, both two enantiomers of dinotefuran have certain toxicity to tadpole cardiomyocytes, while R-dinotefuran exhibited higher toxicity than S-enantiomer, which may be attributed to disparities in the activation capacities of the respiratory chain.