Rapid assessment of heavy metal accumulation capability of Sedum alfredii using hyperspectral imaging and deep learning.

Hyperaccumulators are the material basis and key to the phytoremediation of heavy metal contaminated soils. Conventional methods for screening hyperaccumulators are highly dependent on the time- and labor-consuming sampling and chemical analysis. In this study, a novel spectral approach assisted with multi-task deep learning was proposed to streamline accumulating ecotype screening, heavy metal stress discrimination, and heavy metals quantification in plants. The significant Cd/Zn co-hyperaccumulator Sedum alfredii and its non-accumulating ecotype were stressed by Cd, Zn, and Pb. Spectral images of leaves were rapidly acquired by hyperspectral imaging. The self-designed deep learning architecture was composed of a shallow network (ENet) for accumulating ecotype identification, and a multi-task network (HMNet) for heavy metal stress type and accumulation prediction simultaneously. To further assess the robustness of the networks, they were compared with conventional machine learning models (i.e., partial least squares (PLS) and support vector machine (SVM)) on a series of evaluation metrics of classification, multi-label classification, and regression. S. alfredii with heavy metals accumulation capability was identified by ENet with 100 % accuracy. HMNet reduced overfitting and outperformed machine learning models with the average exact match ratio (EMR) of heavy metal stress discrimination increased by 7.46 %, and residual prediction deviations (RPD) of heavy metal concentrations prediction increased by 53.59 %. The method succeeded in rapidly and accurately discriminating heavy metal stress with EMRs over 91 % and accuracies over 96 %, and in predicting heavy metals accumulation with an average RPD of 3.29 for Zn, 2.57 for Cd, and 2.53 for Pb, indicating the satisfactory practicability and potential for sensing heavy metals accumulation. This study provides a relatively novel spectral method to facilitate hyperaccumulator screening and heavy metals accumulation prediction in the phytoremediation process.

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.

Adsorption and leaching of novel fungicide pyraoxystrobin on soils by 14C tracing method.

Pyraoxystrobin, (E)-2-(2-((3-(4-chlorophenyl)-1-methyl-1H-pyrazole-5-yloxy)methyl)phenyl)-3-methoxyacrylate, is a newly developed strobilurin fungicide with high antifungal efficiency. It has high potential to enter soil environments that might subsequently impact surface and groundwater. Therefore, 14C-labeled pyraoxystrobin was used as a tracer to study the adsorption/desorption and migration behavior of this compound under laboratory conditions in three typical agricultural soils. The adsorption isotherms conformed with the Freundlich equation. Single factor analysis showed that organic matter content was the most important factor influencing the adsorption. The highest adsorption level was measured in soil with low pH and high organic carbon content. Once adsorbed, only 2.54 to 6.41% of the adsorbed compound could be desorbed. In addition, the mobility results from thin-layer chromatography and column leaching studies showed that it might be safe to use pyraoxystrobin as a fungicide without causing groundwater pollution from both runoff and leaching, which might be attributed to its strong hydrophobicity. High organic matter content enhanced pyraoxystrobin adsorption and desorption because of the rule of similarity (lipid solubility). In the column leaching study, 95.02% (minimum value) of the applied 14C remained within the upper 4.0-cm layer after 60 days.

Mineralization and residue characteristics of chloramphenicol in aerobic soils: evidence from a carbon-14 study.

Chloramphenicol, a broad-spectrum antibiotic employed for controlling bacterial infections, presents an intriguing aspect in terms of its environmental fate in soils. 14C-labeled chloramphenicol was used to explore its mineralization and residue characteristics in three distinct agricultural soils in China. The findings revealed a nuanced pattern in the fate of 14C-chloramphenicol, with notable variations among the different soils under investigation. The chloramphenicol extract residue exhibited a reduction of 18.04% in sandy clay soil, 23.04% in clay loam soil, and 21.73% in loamy clay soil. Notably, the mineralization rate in sandy clay soil was 25.22% surpassed that in the other two soils, particularly during the initial stages of incubation. Over time, the diminishing extract residue underwent conversion into minerals and bound residue. The formation rate of bound residue was increased from 44.59 to 53.65% after adding 10% manure, suggesting that chloramphenicol easily binds with soils rich in organic matter. The bound residue is predominantly localized in the humin fraction across all soils. Additionally, the sterilized soil experiments indicated the pivotal role of microorganisms in influencing the fate of chloramphenicol under the specified experimental conditions. In conclusion, this study offers valuable insights into the environmental dynamics of chloramphenicol in soils, emphasizing the importance of soil composition, organic matter content, and microbial activity. The findings contribute to a scientific understanding of the environmental safety implications associated with chloramphenicol usage.

Liver damage and lipid metabolic dysregulation in adult zebrafish (Danio rerio) induced by spirotetramat.

Spirotetramat, an insecticide derived from cycloketone and extensively utilized in agricultural production, has been reported to be toxic to an array of aquatic organisms. Previous studies have indicated that spirotetramat can cause toxicity such as impaired ovarian development and apoptosis in zebrafish, but its toxicological effects on lipid metabolism and liver health in zebrafish remain unclear. In this study, we explored the effects of spirotetramat exposure on zebrafish (Danio rerio) by examining key markers of lipid metabolism, alterations in gene expression related to this process, and histological characteristics of the liver. Spirotetramat significantly reduced the condition factor, triglycerides and low-density lipoprotein cholesterol levels at 2 mg/L. The expression of genes related to fatty acid synthesis (acacb), ß-oxidation (acox1, pparda) and pro-inflammatory cytokines (tnf-α, il-1ß) was downregulated. However, the expression of genes related to lipid transport and uptake (cd36, ppara) and output (apob) was upregulated. The activity of alanine aminotransferase was significantly inhibited. Histopathology results showed that spirotetramat exposure led to liver cell vacuolation and necrosis. In addition, molecular docking results of spirotetramat and lipid transport related protein (ACC, ApoB) in both zebrafish and human showed the binding energy of human proteins is lower than that for zebrafish, and that the number of hydrogen bonds formed was higher. It is speculated that spirotetramat may also pose a significant potential hazard to humans, potentially affecting human lipid metabolism and health. This study expunge shed light on the ecological toxicity of spirotetramat by showing how it disrupts lipid metabolism and causes tissue damage specifically in zebrafish liver, contributing to a deeper understanding of its harmful effects in aquatic environment.

Plant Essential Oils: Dual Action of Toxicity and Egg-Laying Inhibition on Tetranychus urticae (Acari: Tetranychidae), Unveiling Their Potential as Botanical Pesticides.

Tetranychus urticae, a prominent pest mite in strawberry and vegetable cultivation in China, has developed escalating resistance due to extensive chemical pesticide application. Consequently, there is an urgent need to identify safe and efficacious methods to reduce resistance development. In this study, 38 commercially available plant essential oils (EOs) were screened for their acaricidal potential and ability to inhibit oviposition. The findings revealed that 13 EOs exhibited notable acaricidal activity, with lemon EO demonstrating the highest toxicity, followed by sage, patchouli, frankincense, lemongrass, palmarosa, and oregano EOs. In addition, 18 EOs displayed significant inhibitory effects on oviposition, with lemon EO exhibiting the highest inhibition rate (99.15%) and inhibition index (0.98). Subsequently, sage, frankincense, clove, lemongrass, oregano, patchouli, myrrh, black pepper, palmarosa, and geranium EOs also showed inhibition rates exceeding 50%. Despite black pepper, clove, myrrh, and oregano EOs demonstrating relatively low toxicity against T. urticae, they exhibited heightened efficacy in inhibiting oviposition and suppressing population expansion. This study conducted a comparative assessment of the acaricidal and oviposition inhibition activities of EOs and their principal constituents, thus providing a theoretical basis for the development of botanical acaricides against T. urticae.

Structure and physicochemical properties of starches from six accessions of the genus Pueraria in China.

Kudzu (Pueraria lobata) root contains abundant starch, but the physicochemical properties of kudzu starch are not well understood. In this study, we compared the compositions and physicochemical properties of starches isolated from six Pueraria accessions in China. Caige starch exhibited the highest purity (96.99 %) and amylose content (24.76 %), while Yege starch contained higher levels of puerarin (493.37 µg/g) and daidzein (38.68 µg/g). All kudzu starches were rich in resistant starch, with RS2 content ranging from 38.61 % to 46.22 % and RS3 content from 3.59 % to 6.04 %. The granules of kudzu starches varied in morphology, with Yege starch featuring larger polygonal granules. The kudzu starches presented either A-type or A-type-like C-type diffraction patterns. Caige starch had a higher IR2 value (1.28), higher gelatinization temperatures, wider temperature ranges, and greater enthalpy changes. Yege (JX) starch exhibited the highest peak viscosity but the lowest setback viscosity and pasting temperature. Fenge starch showed the highest final viscosity, with Fenge (ZJ) starch demonstrating the highest crystallinity (25.7 %) and IR1 value (0.80). These results indicated that kudzu starches derived from various Pueraria species possess unique structural and physicochemical properties, which provide significant potential for applications in food and other industrial fields.

C-Terminal region of DNA ligase IV drives XRCC4/DNA ligase IV complex to chromatin.

DNA ligase IV (LIG4) and XRCC4 form a complex to ligate two DNA ends at the final step of DNA double-strand break (DSB) repair through non-homologous end-joining (NHEJ). It is not fully understood how these proteins are recruited to DSBs. We recently demonstrated radiation-induced chromatin binding of XRCC4 by biochemical fractionation using detergent Nonidet P-40. In the present study, we examined the role of LIG4 in the recruitment of XRCC4/LIG4 complex to chromatin. The chromatin binding of XRCC4 was dependent on the presence of LIG4. The mutations in two BRCT domains (W725R and W893R, respectively) of LIG4 reduced the chromatin binding of LIG4 and XRCC4. The C-terminal fragment of LIG4 (LIG4-CT) without N-terminal catalytic domains could bind to chromatin with XRCC4. LIG4-CT with W725R or W893R mutation could bind to chromatin but could not support the chromatin binding of XRCC4. The ability of C-terminal region of LIG4 to interact with chromatin might provide us with an insight into the mechanisms of DSB repair through NHEJ.

The antifungal potential of the chelating agent EDTA against postharvest plant pathogen Botrytis cinerea.

Botrytis cinerea is a phytopathogenic fungus that causes gray mold, a major postharvest disease of fruits and vegetables. Chemical fungicides remain the main solution to control Botrytis disease, but concerns have raised about their safety to environment and human health, and there is an increasing need for development of more effective and less toxic treatments. In this study the divalent cation chelating agent ethylenediaminetetraacetic acid (EDTA) exhibited marked antifungal activity against B. cinerea, including inhibition of spore germination, mycelial growth, infection cushion formation, stimulation of cell death, and impairment of fungal virulence. These adverse effects of EDTA could be reversed by the addition of calcium ion, implying that metal ion chelation is involved in the fungicidal mechanism. Bean leaf and tomato fruit protection assay indicated that EDTA treatment led to a significant reduction of infection by B. cinerea. Furthermore, the antifungal activity of EDTA was significantly enhanced when used in combination with fenhexamid. These findings suggest that EDTA could be a promising tool to control B. cinerea, and application of EDTA may reduce the use of conventional chemical fungicides.

[Effects of Low-density Polyethylene Microplastics on the Growth and Physiology Characteristics of Ipomoea aquatica Forsk].

Microplastic pollution in soil and its toxicological effects have attracted increasing attention from researchers, but the mechanisms of microplastics affecting crop growth and physiology remain unclear. A pot experiment was conducted to evaluate the impacts of various mass concentrations (0%, 0.2%, 5%, and 10%) of low-density polyethylene microplastics (LDPE MPs) on the germination rate, photosynthetic pigment content, biomass, antioxidant enzyme activity, soluble protein, and soluble sugar content of water spinach (Ipomoea aquatica Forsk). The results showed that LDPE MPs significantly inhibited (P<0.05) the seed vigor of water spinach, and the inhibitory effect increased with increasing concentration of LDPE MPs. However, the 5% LDPE MPs significantly promoted the aboveground biomass of water spinach. The 0.2% and 10% LDPE MPs significantly improved the superoxide dismutase (SOD) activity and catalase (CAT) and peroxidase (POD) activities, respectively. Further, malondialdehyde (MDA) content decreased with increasing concentration of LDPE MPs, and the reductions reached 15.53%-27.39% in comparison to that in the control. The LDPE MPs also significantly increased the soluble sugar content of water spinach leaves. In summary, LDPE MPs could inhibit the seed vigor and promote biomass accumulation in water spinach. Water spinach could relieve the oxidative stress caused by LDPE MPs by regulating antioxidant enzyme activity and soluble protein content. Therefore, this study may provide basic information for assessing the influences of microplastics on vegetables.

Embryonic exposure to prothioconazole induces oxidative stress and apoptosis in zebrafish (Danio rerio) early life stage.

Triazole fungicides are extensively applied in general agriculture for fungal control and have negative impacts on aquatic organisms. Prothioconazole, a widely used triazole fungicide, is toxic to zebrafish, but systematic research on the negative effects caused by prothioconazole in zebrafish embryos is limited. In this study, we studied the developmental toxicology, oxidative stress and apoptosis caused by prothioconazole in zebrafish embryos. Exposure to 0.850 mg/L prothioconazole impacts embryo survival and hatching. Prothioconazole exposure caused embryo malformation, especially yolk-sac and pericardial edemas, and prothioconazole-induced apoptosis was observed. Additionally, exposure to a high prothioconazole concentration up-regulated the expression levels of oxidative stress defense-related genes and p53. The bax to bcl2 ratio increased along with exposure time and prothioconazole concentration. Prothioconazole induced apoptosis during the early life stages of zebrafish and may trigger oxidative-stress and p53-dependent pathway responses. Our findings increase our understanding of the molecular mechanisms of oxidative stress and cell death caused by prothioconazole.

Comparative Transcriptome Analysis Reveals the Mechanism Related to Fluazinam Stress of Panonychus citri (Acarina: Tetranychidae).

The use of a large number of chemical acaricides to control these pest mites has led to an increasing problem of pesticide resistance, which has always been the difficulty in integrated pest management (IPM). Fluazinam has a good control effect on Panonychus citri, the serious pest on citrus; however, we only know the mechanism of action of fluazinam as a fungicide and its mechanism of action on mites remains unclear. Through analysis using Illumina high-throughput transcriptomic sequencing and differential expression genes in P. citri treated with fluazinam, 59 cytochrome P450 genes, 23 glutathione s-transferase genes, five carboxylate esterase genes, 11 superoxide dismutase genes and 15 catalase genes were identified. The Gene Ontology enrichment and the enrichment of KEGG results showed that the treatment were enrichment for redox enzyme pathways. Evaluating the efficacy of fluazinam, and analyzing the transcriptome data of P. citri under fluazinam stress, potentially provide a new agent for prevention and control of P. citri, and also preliminary research results for exploring the mechanism of action of fluazinam on P. citri. Given the up-regulated expression levels of genes for Mn-superoxide dismutase and catalase, we speculate that they play an important role in fluazinam-stress action on P. citri.

Phylogenetic analyses and characteristics of the microbiomes from five mealybugs (Hemiptera: Pseudococcidae).

Associations between Sternorrhyncha insects and intracellular bacteria are common in nature. Mealybugs are destructive pests that seriously threaten the production of agriculture and forestry. Mealybugs have evolved intimate endosymbiotic relationships with bacteria, which provide them with essential amino acids, vitamins, and other nutrients. In this study, the divergence of five mealybugs was analyzed based up the sequences of the mitochondrial cytochrome oxidase I (mtCOI). Meanwhile, the distinct regions of the 16S rRNA gene of primary symbionts in the mealybugs were sequenced. Finally, high-throughput sequencing (HTS) techniques were used to study the microbial abundance and diversity in mealybugs. Molecular phylogenetic analyses revealed that these five mealybugs were subdivided into two different clusters. One cluster of mealybugs (Dysmicoccus neobrevipes, Pseudococcus comstocki, and Planococcus minor) harbored the primary endosymbiont "Candidatus Tremblaya princeps," and another cluster (Phenacoccus solenopsis and Phenacoccus solani) harbored "Ca. Tremblaya phenacola." The mtCOI sequence divergence between the two clusters was similar to the 16S rRNA sequence divergence between T. princeps and T. phenacola. Thus, we concluded that the symbiont phylogeny was largely concordant with the host phylogeny. The HTS showed that the microbial abundance and diversity within P. solani and P. solenopsis were highly similar, and there was lower overall species richness compared to the other mealybugs. Among the five mealybugs, we also found significant differences in Shannon diversity and observed species. These results provide a theoretical basis for further research on the coevolution of mealybugs and their symbiotic microorganisms. These findings are also useful for research on the effect of symbiont diversity on the pest status of mealybugs in agricultural systems.

Plant Defense Responses Induced by Two Herbivores and Consequences for Whitefly Bemisia tabaci.

Diverse herbivores are known to induce various plant defenses. The plant defenses may detrimentally affect the performance and preference to subsequent herbivores on the same plant, such as affecting another insect's feeding, settling, growth or oviposition. Here, we report two herbivores (mealybug Phenacoccus solenopsis and carmine spider mite Tetranychus cinnabarinus) which were used to pre-infest the cucumber to explore the impact on the plants and the later-colonizing species, whitefly Bemisia tabaci. The results showed that the whiteflies tended to select the treatments pre-infested by the mites, rather than the uninfected treatments. However, the result of treatments pre-infested by the mealybugs was opposite. Total number of eggs laid of whiteflies was related to their feeding preference. The results also showed that T. cinnabarinus were more likely to activate plant jasmonic acid (JA) regulated genes, while mealybugs were more likely to activate key genes regulated by salicylic acid (SA). The different plant defense activities on cucumbers may be one of the essential factors that affects the preference of B. tabaci. Moreover, the digestive enzymes and protective enzymes of the whitefly might play a substantial regulatory role in its settling and oviposition ability.

Uptake and distribution characteristics of the novel fungicide pyraoxystrobin in cucumber plants.

The uptake and distribution characteristics of a novel fungicide, pyraoxystrobin, labeled with 14C on its pyrazol ring, were investigated in cucumber (Cucumis sativus) seedlings. Foliar applied pyraoxystrobin rapidly penetrated the treated leaf and reached a maximum uptake of 68% after 5 d. The translocation of absorbed 14C in cucumber seedlings was both acropetal and basipetal. However, over 74% of the absorbed 14C-pyraoxystrobin remained in the treated leaves. The order of its distribution in the plant was as follows: treated leaf > stalk above the treated leaf > leaves above the treated leaf > stalk below the treated leaf > leaves below the treated leaf > cotyledon > root. Seedlings grown in soils containing bound residues (BR) of pyraoxystrobin revealed that the BRs were not easily absorbed or translocated by the plant. Soil type had a large effect on root uptake, with the highest uptake among the three tested soils from red clay.

Adsorption of uranium by amidoximated chitosan-grafted polyacrylonitrile, using response surface methodology.

The amidoximated chitosan-grafted polyacrylonitrile (CTS-g-PAO) was prepared for the adsorption of uranium from water. The effects of pH, concentration of uranium and the solid-liquid ratio on the adsorption of uranium by CTS-g-PAO were optimized using Doehlert design of response surface methodology (RSM). The adsorption capacity and removal efficiency achieved 312.06 mg/g and 86.02%, respectively. The adsorption process attained equilibrium only in 120 min. More than 80% of the absorbed uranium could be desorbed by 0.1 mol/l HCl or EDTA-Na, and CTS-g-PAO could be reused at least 3 times. The CTS-g-PAO and U(VI) ions formed a chelate complex due to FTIR spectral analysis. The surface morphology of CTS-g-PAO was also investigated by SEM. The adsorption process was better described by Langmuir isotherm and pseudo second order kinetic model. Results obtained indicated that CTS-g-PAO was very promising in adsorption of uranium from water.

Fate of a novel strobilurin fungicide pyraoxystrobin in flooded soil.

Pyraoxystrobin, ((E)-2-(2-((3-(4-chlorophenyl)-1-methyl-1H-pyrazol-5-yloxy) methyl) phenyl)-3-methoxyacrylate) is a novel strobilurin fungicide with excellent and broad spectrum antifungal efficiency. Environmental behaviors of the new fungicide must be assessed to understand its potential risks to the environment. In this study, the extractable residues, bound residues and mineralization, as well as the dissipation rates of pyraoxystrobin were investigated in three flooded soils using a (14)C tracing technique. Results showed that pyraoxystrobin didn't undergo appreciable dissipation during the 100 day incubation period in some tested soils, with 70.01%, 28.58% and 83.85% of the parent compound remaining in the solonchak, cambisol and acrisol soils at the end of the experiment, respectively. Almost no (14)C-pyraoxystrobin was mineralized to (14)CO2 (<0.5%) over the experimental period. Organic matter had a dominating influence on the bound residues formation and the fractions of bound residues increased as the soil organic matter content increased. Less than 9% of the radioactivity was found in the aqueous phase, while the majority of extractable residues (>65.39%) were recovered in the organic extracts. This study aims to give a deep insight into the environmental behaviors of pyraoxystrobin and may be beneficial for the risk assessment of other analogous fungicides.

Dynamic characteristics of the novel strobilurin fungicide SYP-3343 in aerobic soils.

SYP-3343, (E)-2-(2-((3-(4-chlorophenyl)-1-methyl-1H-pyrazole-5-yloxy)methyl)phenyl)-3-methoxyacrylate, is a newly developed strobilurin fungicide. However, the environmental behavior and fate of SYP-3343 in soil have not been well-documented. In this study, ¹4C-labeled SYP-3343 was employed to investigate the dynamic characteristics in three typical soils under aerobic conditions. Radioactivity analysis after high-performance liquid chromatography (HPLC) showed that SYP-3343 degraded rapidly in the coastal soil with a half-life of 43.8 days. After incubation of 100 days, its extractable residues were greater than 76.0% and bound residues were less than 12.4%, indicating that SYP-3343 was not easy to accumulate in soils. The mineralization to ¹4CO2 reached 5.4% for acidic soil, 2.8% for neutral soil, and 1.7% for alkaline soil, suggesting that it was difficult to cleave the pyrazole ring completely. In addition, dynamic characteristics of SYP-3343 in sterile and non-sterile loamy soil showed that soil microbes affected SYP-3343 residue in soil and could accelerate the process of degradation and mineralization.