Physiological and biochemical regulation of tobacco by oxathiapiprolin under Phytophthora nicotianae infection.

As a fungicide, oxathiapiprolin has excellent effects on diseases caused by oomycetes. Fungicides generally protect crops by inhibiting pathogens, but little research has addressed the effects of fungicides on crops. This study combined transcriptomic and metabolomic analyses to systematically analyze the physiological regulatory mechanisms of oxathiapiprolin on tobacco under Phytophthora nicotianae infection. The results showed that under P. nicotianae infection, tobacco's photosynthetic rate and antioxidant enzyme activity increased after the application of oxathiapiprolin. Omics results showed that the genes related to carbon metabolism, disease-resistant proteins, and amino acid synthesis were highly expressed, and the amino acid content increased in tobacco leaves. This study is the first comprehensive investigation of the physiological regulatory effects of oxathiapiprolin on tobacco in response to P. nicotianae infection. These findings provide a basis for the balance between regulating tobacco growth and development and enhancing disease resistance under the stimulation of oxathiapiprolin and provide new research and development opportunities for identifying new disease-resistance genes and the development of high-yielding disease-resistant crop varieties.

Direct toxicity of the herbicide florasulam against Chlorella vulgaris: An integrated physiological and metabolomic analysis.

Herbicides are the agents of choice for use in weed control; however, they can enter the aquatic environment, with potentially serious consequences for non-target organisms. Despite the possible deleterious effects, little information is available regarding the ecotoxicity of the herbicide florasulam toward aquatic organisms. Accordingly, in this study, we investigated the toxic effect of florasulam on the freshwater microalga Chlorella vulgaris and sought to identify the underlying mechanisms. For this, we employed a growth inhibition toxicity test, and then assessed the changes in physiological and metabolomic parameters, including photosynthetic pigment content, antioxidant system, intracellular structure and complexity, and metabolite levels. The results showed that treatment with florasulam for 96 h at the concentration of 2 mg/L, 2.84 mg/L, and 6 mg/L in medium significantly inhibited algal growth and photosynthetic pigment content. Moreover, the levels of reactive oxygen species were also increased, resulting in oxidative damage and the upregulation of the activities of several antioxidant enzymes. Transmission electron microscopic and flow cytometric analysis further demonstrated that exposure to florasulam (6 mg/L) for 96 h disrupted the cell structure of C. vulgaris, characterized by the loss of cell membrane integrity and alterations in cell morphology. Changes in amino acid metabolism, carbohydrate metabolism, and the antioxidant system were also observed and contributed to the suppressive effect of florasulam on the growth of this microalga. Our findings regarding the potential risks of florasulam in aquatic ecosystems provide a reference for the safe application of this herbicide in the environment.

Oxidative stress and detoxification mechanisms of earthworms (Eisenia fetida) after exposure to flupyradifurone in a soil-earthworm system.

Flupyradifurone (FLU) has great application potential in agricultural production as a new generation of neonicotinoid insecticide after imidacloprid. Nevertheless, the toxic effects of FLU on non-target soil organisms remain unclear, resulting in considerable environmental risks. We evaluated the acute and subchronic toxicities of FLU to earthworms. The results of acute toxicity show that the median lethal concentration (LC50) values (14 d) of FLU were 186.9773 mg kg-1 for adult earthworms and 157.6502 mg kg-1 for juveniles, respectively. The subchronic toxicity of FLU that focused on the activities of antioxidant and detoxication enzymes showed the superoxide dismutase (SOD), catalase (CAT), and glutathione-S transferase (GST) activities in earthworms increased while the peroxidase (POD) and acetylcholinesterase (AChE) activities decreased after exposure to FLU. Oxidative damage analyses revealed that the reactive oxygen species (ROS) level and malonaldehyde (MDA) content in earthworms were increased by FLU, resulting in DNA damage. Transcriptomics and RT-qPCR confirmed that FLU influenced the expression of genes related to antioxidant response and detoxification of earthworms. Ultimately detoxification metabolism, environmental information processing, cell processes, and immune system pathways are significantly enriched to respond jointly to FLU. Our study fills the gaps in the toxicity of FLU to earthworms, providing a basis for its risk assessment of soil ecosystems and non-target biological toxicity.

Transcriptomics and enzymology combined five gene expressions to reveal the responses of earthworms (Eisenia fetida) to the long-term exposure of cyantraniliprole in soil.

Cyantraniliprole is a novel diamide insecticide that acts upon the ryanodine receptor (RyR) and has broad application prospects. Accordingly, it is very important to evaluate the toxicity of cyantraniliprole to earthworms (Eisenia fetida) because of their vital role in maintaining a healthy soil ecosystem. In this study, an experiment was set up, using four concentrations (0.1, 1, 5, and 10 mg/kg) and solvent control group (0 mg/kg), to investigate the ecotoxicity of cyantraniliprole to earthworms. Our results showed that, after 28 days of exposure to cyantraniliprole, both cocoon production and the number of juvenile earthworms had decreased significantly at concentrations of either 5 or 10 mg/kg. On day 14, we measured the activities of digestive enzymes and ion pumps in the intestinal tissues of earthworms. These results revealed that cyantraniliprole exposure caused intestinal damage in earthworm, specifically changes to its intestinal enzyme activity and calcium ion content. Cyantraniliprole could lead to proteins' carbonylation under the high-dose treatments (i.e., 5 mg/kg, 10 mg/kg). At the same time, we also found that cyantraniliprole can cause the abnormal expression of key functional genes (including HSP70, CAT, RYR, ANN, and CAM genes). Moreover, the transcriptomics data showed that exposure to cyantraniliprole would affect the synthesis of carbohydrates, proteins and lipids, as well as their absorption and transformation, while cyantraniliprole would also affect signal transduction. In general, high-dose exposure to cyantraniliprole causes reproductive toxicity, genotoxicity, and intestinal damage to earthworms.

Responses of Harmonia axyridis (Coleoptera: Coccinellidae) to sulfoxaflor exposure.

Harmonia axyridis is an important predator of several pest species and is part of many Integrated Pest Management (IPM) programs. To assess the risks of pesticide application to H. axyridis, we studied the effects of sulfoxaflor on H. axyridis larvae. At 72 h after treatment, the acute toxicity LR50 was 311.9476 g a. i. ha-1 by the residual contact method. This result indicated low-contact toxicity against second-instar H. axyridis larvae. The LR50 of the F1 generation decreased from 69.96 to 36.41 g a. i. ha-1 in a long-term toxicity test. The daily hazard quotient (HQ) for H. axyridis larvae lowered the safety threshold value in the first 5 d. However, the HQ values were greater than 2 during days 6-18 after sulfoxaflor treatments. We determined the No Observed Effect Application Rates of sulfoxaflor on the survival (<11.25 g a. i. ha-1), duration of larval and pupal stages (45 g a. i. ha-1), adult stage (90 g a. i. ha-1), total pre-oviposition period, adult pre-oviposition period (45 g a. i. ha-1), and reproduction (11.25 g a. i. ha-1). Pupation, adult emergence, and eggs counts of H. axyridis were reduced after sulfoxaflor treatments. The predation ability and population demography parameters were significantly impaired by higher application rates. At 90 g a. i. ha-1 or less, sulfoxaflor was slightly harmful to H. axyridis but a rate of 180 g a. i. ha-1 was moderately harmful. These results demonstrated that sulfoxaflor is harmful to H. axyridis when applied at high application rates.

Toxicity of thifluzamide in earthworm (Eisenia fetida).

An increase in the area treated with the fungicide thifluzamide has triggered concerns for soil ecosystem service providers such as earthworms. Here, we assessed effects of thifluzamide on earthworm (Eisenia fetida) biomarker indicators of stress responses and reproduction following exposure to 0, 0.1, 1.0, and 10.0 mg of thifluzamide kg-1 soil for 7, 14, 21, and 28 d (biomarker indicators) and 30 d (reproduction). Growth and reproduction were inhibited by exposure to thifluzamide at 10.0 mg/kg, and the activities of succinate dehydrogenase (SDH) and respiratory chain complex II were inhibited by exposure to 1.0 and 10.0 mg/kg thifluzamide for the majority of the 28-d experiment. Reactive oxygen species (ROS) increased across all thifluzamide treatments, and the activities of superoxide dismutase (SOD) and glutathione-S-transferase (GST) tended to be inhibited by thifluzamide. Upon exposure to thifluzamide, the activities of catalase (CAT) and guaiacol peroxidase (POD) initially increased and then decreased. Increased levels of malondialdehyde (MDA) were detected only at seven days after exposure, and genotoxicity increased as the thifluzamide concentration increased. The results suggest that thifluzamide presents a potential risk to earthworms at the concentration of 10.0 mg/kg, and its use should be moderated to reduce damage to soil ecosystem function.

Effects of insect growth-regulator insecticides on the immature stages of Harmonia axyridis (Coleoptera: Coccinellidae).

Harmonia axyridis is an important biological control predator in greenhouses and agricultural fields, and it plays a significant role in the integrated pest management (IPM) of several arthropod pests. We studied the effects of eight insect growth-regulator insecticides (IGRs) on immature stages of H. axyridis by residual toxicity (eggs and pupae) and contact toxicity (larvae) to evaluate the risk of using these IGRs in IPM systems. Diflubenzuron, hexaflumuron and lufenuron caused more than 80% mortality to H. axyridis eggs, larvae and pupae, respectively. Pyriproxyfen was also highly harmful to larvae and pupae of H. axyridis. In contrast, methoxyfenozide and buprofezin caused little mortality and were classified as slightly harmful to immature stages based on a reduction coefficient. In addition to mortality and developmental time, the fecundity, fertility and deformed eggs of offspring were affected, when the predators were exposed to IGRs. Benzoylphenylurea insecticides significantly reduced H. axyridis female fecundity and fertility and increased the number of deformed eggs. The adverse effects are closely connected with the developmental stages of the predators and types and methods of insecticides exposed. All IGRs affected, to some extent, the life-table parameters of H. axyridis when the insecticides applied on immature stages at the highest field rates. Tebufenozide, diflubenzuron, hexaflumuron and lufenuron significantly reduced the Ro, T, r and λ of beetles exposed to the insecticides. The results indicate that IGRs could disturb the population growth and biocontrol activities of H. axyridis when applied at the highest field label rates. Additional studies should be conducted to assess the effects of IGRs on H. axyridis under field conditions before incorporating them in IPM strategies.

Cyantraniliprole seed treatment effectively controls wireworms (Pleonomus canaliculatus Faldermann) and white grubs (Anomala corpulenta Motschulsky) in maize fields.

Wireworms and white grubs are destructive underground pests in maize fields in China. Cyantraniliprole has good control effect on coleoptera pests. Here, we evaluated the toxicity of cyantraniliprole to the second instar larvae of Anomala corpulenta Motschulsky and third-instar of larvae of Pleonomus canaliculatus Faldermann and the effects of sublethal concentrations on the activity of antioxidant and detoxification enzymes. We also explored the efficacy of cyantraniliprole on underground pests under indoor and field conditions. The LC50 of cyantraniliprole for the third instar larvae of P. canaliculatus was 23.3712 mg/L, and that for the second instar larvae of A. corpulenta was 5.9715 mg/L. Cyantraniliprole can activate the activity of superoxide dismutase (SOD), peroxidase (POD), and glutathione S-transferase (GST) to different degrees at a sublethal dose. According to the pot experiment and the control efficacy test in the field, the indoor control effect of cyantraniliprole seed treatment on P. canaliculatus and white grubs was approximately 80%, and the maximum increase in yield achieved through cyantraniliprole application was approximately 15% in the field efficacy test. Cyantraniliprole has a strong control effect on wireworms and white grubs, so it can be used to treat seeds to control underground pests in maize fields.

Toxic effects of fludioxonil on the growth, photosynthetic activity, oxidative stress, cell morphology, apoptosis, and metabolism of Chlorella vulgaris.

Fludioxonil is widely used in the control of crop diseases because of its broad spectrum and high activity, but its presence is now common in waterways proximate to treated areas. This study examined the toxic effects and mechanisms of fludioxonil on the microalgal taxa Chlorella vulgaris. The results showed that fludioxonil limited the growth of C. vulgaris and the median inhibitory concentration at 96 h was 1.87 mg/L. Concentrations of 0.75 and 3 mg/L fludioxonil reduced the content of photosynthetic pigments in algal cells to different degrees. Fludioxonil induced oxidative damage by altering C. vulgaris antioxidant enzyme activities and increasing reactive oxygen species levels. Fludioxonil at 0.75 mg/L significantly increased the activity of antioxidant enzymes. The highest level of activity was 1.60 times that of the control group. Both fludioxonil treatment groups significantly increased ROS levels, with the highest increase being 1.90 times that of the control group. Transmission electron microscope showed that treatment with 3 mg/L fludioxonil for 96 h disrupted cell integrity and changed cell morphology, and flow cytometer analysis showed that fludioxonil induced apoptosis. Changes in endogenous substances indicated that fludioxonil negatively affects C. vulgaris via altered energy metabolism, biosynthesis of amino acids, and unsaturated fatty acids. This study elucidates the effects of fludioxonil on microalgae and the biological mechanisms of its toxicity, providing insights into the importance of the proper management of this fungicide.

Effects of thifluzamide on soil fungal microbial ecology.

Thifluzamide, a succinate dehydrogenase inhibitor fungicide, has been used extensively for many diseases control and has the risk of accumulation in soil ecology. In order to study the ecotoxicity of thifluzamide to soil fungal communities, typical corn field soils in north (Tai'an) and south (Guoyang) China were treated with thifluzamide (0, 0.1, 1.0 and 10.0 mg/kg) and incubated for 60 days. Thifluzamide exposure promoted soil basal respiration, and significantly reduced the number of soil culturable fungi and the abundance of soil fungi (RT-qPCR) in middle and late treatment period (15, 30, 60 days). Illumina Mi-Seq sequencing revealed that thifluzamide could reduce fungal alpha diversity (Sobs, Shannon, Simpson indexes) and change fungal community structure. FUN Guild analysis showed that the relative abundance of Undefined Saprotroph increased after the thifluzamide treatment, whereas that of Plant Pathogen decreased, and we concluded that exposure to thifluzamide could change the function of soil fungi. This study evaluated the soil ecological risk caused by thifluzamide's release into soil, providing a basis for its rational application.

Effects of a novel fungicide benzovindiflupyr in Eisenia fetida: Evaluation through different levels of biological organization.

Although benzovindiflupyr (BZF), which is a novel succinate dehydrogenase inhibitor fungicide, has considerable application potential worldwide, its extensive use is toxic to non-targeted soil organisms. Therefore, this study aimed to evaluate the acute and subchronic toxicity of BZF to earthworms (Eisenia fetida). The acute toxicity of BZF to adult and larval earthworms was measured, as indicated by the following LC50 values obtained after 14 days of exposure: 416 mg/kg for adult earthworms and 341 mg/kg for juveniles. Subchronic toxicity tests were conducted using only adult earthworms. The earthworms' weight gain was slower on days 14 and 28 after commencing the BZF T100 treatment (50 mg/kg of soil). Following 14 days of BZF exposure, enzymes and gene expressions associated with the mitochondrial respiratory chain and energy metabolism were activated to some extent, and the reactive oxygen species level and malondialdehyde content also increased. Antioxidant and detoxifying enzymes and metallothionein gene, Heat shock protein 70 gene associated with resistance to oxidative damage were also activated to varying degrees. Increased BZF concentrations corresponded to increased genotoxicity. Integrated biological response (IBR) values were calculated at the biochemical and molecular levels to show increased toxicity with increased BZF concentration. Although a series of biomarkers changes occurred after initiating BZF treatment, these changes were all likely to have been resisted by the earthworms' own antioxidant defense system and only showed phenotypic (weight-related) changes with treatments of 50 mg/kg. In conclusion, reasonable levels of BZF application may have little impact on earthworms. Our findings provide insights on the toxic effects of BZF on earthworms and may prove useful for risk assessments relating to BZF's impacts on soil ecosystems.

Effects of mesotrione on oxidative stress, subcellular structure, and membrane integrity in Chlorella vulgaris.

Mesotrione is a selective herbicide used to prevent weed attack of corn. It is extensively used, and hence, is being increasingly detected in aquatic ecosystems and may exert adverse effects on aquatic organisms. To evaluate the effects of mesotrione on photosynthesis-related gene expression, antioxidant enzyme activities, subcellular structure, and membrane integrity in algal cells, a comprehensive study was conducted using the green alga, Chlorella vulgaris. Exposure to 4-50 mg/L mesotrione resulted in a progressive inhibition of cell growth, with a 96-h median inhibition concentration (96 h- ErC50) value of 18.8 mg/L. Further, 18 and 37.5 mg/L mesotrione affected the algal photosynthetic capacity by decreasing the cell pigment content and reducing transcript abundance of photosynthesis-related genes. Mesotrione induced oxidative stress, as confirmed by increased cellular levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and altered antioxidant enzyme activities. It also damaged the algal cellular structure, observed as plasmolysis, blurred organelle shape, and disruption of the chloroplast structure. Flow cytometry analysis revealed that mesotrione exposure led to uneven cell growth and interior irregularities in the algal cell. The apparent propidium iodide (PI) influx also confirmed that the herbicide induced damage of the cell membrane integrity. This study will facilitate the understanding of the physiological and morphological changes induced by mesotrione in C. vulgaris cells, and provide basic information for understanding the biological mechanisms of mesotrione-induced algal toxicity.

Growth, DNA damage and biochemical toxicity of cyantraniliprole in earthworms (Eisenia fetida).

Cyantraniliprole is a second-generation diamide insecticide that exhibited excellent biological efficacy against a variety of pests. To assess the toxic impact of cyantraniliprole on earthworms, the levels of reactive oxygen species (ROS) and malondialdehyde (MDA), activities of superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST), as well as DNA damage were measured after exposed to five cyantraniliprole concentrations ranging from 0 to 10.00 mg/kg for 7, 14, 21 and 28 days. In most treatment groups, the ROS levels increased significantly before exposure time of 14 days and then returned to normal levels. However, the SOD and CAT activities showed different response with activities were first significantly decreased and subsequently increased. The peroxidase (POD) activity showed no significant differences between treatment and control groups at first and then significantly increased. However, the opposite pattern characterized the GST activity. Also, maybe being dose-dependent before 14 days. The MDA concentration was used as a measure of lipid peroxidation (LPO). During experiment period, the MDA concentrations significantly increased when treated by this pesticide. The olive tail moment (OTM) was used as a measure of DNA damage. At higher concentrations of cyantraniliprole and longer exposure times, the OTM gradually increased, and DNA damage in the earthworms gradually increased. The weight of the high-dose (i.e., 5 mg/kg, 10 mg/kg) earthworms showed a significant trend of decrease phenomenon. Overall, the results suggest that sub-chronic exposure to cyantraniliprole causes DNA damage and LPO, weight loss and growth inhibition, leading to antioxidant defence responses in earthworms.

Chlorantraniliprole against the black cutworm Agrotis ipsilon (Lepidoptera: Noctuidae): From biochemical/physiological to demographic responses.

Agrotis ipsilon (Lepidoptera: Noctuidae) is a major underground pest that damages many agricultural crops in China and other countries. A diet-incorporation-based bioassay was conducted to evaluate the sublethal effects of the novel anthranilic diamide chlorantraniliprole on the nutritional physiology, enzymatic properties and population parameters of this cutworm. Chlorantraniliprole exhibited signs of active toxicity against third instar larvae of A. ipsilon, and the LC50 was 0.187 µg.g-1 of artificial diet after treatment for 72 h. The development time of the larval, pupal and adult stages was significantly affected after chlorantraniliprole exposure, compared to the control treatment. Relative to the control treatment, chlorantraniliprole decreased pupal and adult emergence rates, fecundity and fertility and increased the proportions of developmental deformities, the adult preoviposition period (APOP) and the total preoviposition period (TPOP). Furthermore, compared to those treated with the control, A. ipsilon larvae treated with low doses of chlorantraniliprole decreased food utilization and nutrient content (protein, lipid, carbohydrate, trehalose), showed lower pupal weights and growth rates. Compared with the control treatment, chlorantraniliprole significantly reduced digestive enzyme activities and observably increased detoxifying and protective enzyme activities and hormone titers. Importantly, these chlorantraniliprole-induced changes affected life table parameters of the cutworm. These results suggest that chlorantraniliprole at low concentrations can impair A. ipsilon development duration, normal food consumption and digestion process, enzymatic properties, hormone levels, fecundity and population levels. Chlorantraniliprole exhibit the potential to be exploited as a control strategy for this cutworm.

Compatibility of chlorantraniliprole with the generalist predator Coccinella septempunctata L. (Coleoptera: Coccinellidae) based toxicity, life-cycle development and population parameters in laboratory microcosms.

Coccinella septempunctata is a common insect predator that provides biological control of many destructive arthropod pests. An assessment of the toxicity of pesticides to predators is a necessary component of Integrated Pest Management (IPM) strategies. In order to evaluate the risks of field insecticide application, we studied the influence of chlorantraniliprole on C. septempunctata larvae using different exposure doses. Chlorantraniliprole exhibited low contact toxicity against 2nd instar larvae of C. septempunctata with the LR50 was 482.7063 g a.i. ha-1 by after a 72-h exposure. In a long-term test, the LR50 of chlorantraniliprole for C. septempunctata decreased from 88.97 to 58.22 g a.i. ha-1, while the hazard quotient (HQ) values were below the threshold value of 2 during the entire observation period. This indicated a low toxicity risk from insecticide exposure. The total effect (E) suggested that chlorantraniliprole could be classified as harmless/slightly harmful to C. septempunctata below/at 200% of the MRFR (the manufacturer maximum recommended field rate) of 120 g a.i. ha-1. We also determined no observed effect application rates (NOERs) of chlorantraniliprole on survival (7.5 g a.i. ha-1), developmental time (15 g a.i. ha-1) and fecundity (30 g a.i. ha-1). Chlorantraniliprole significantly reduced the pupation rate, adult emergence, egg hatchability, and predation success. Population parameters, including R0, r, λ, and T were significantly affected when C. septempunctata were treated with chlorantraniliprole at higher label rates. These results demonstrated that the use of chlorantraniliprole may reduce C. septempunctata population levels and the level of biological control provided by this species.

[Effects of seed dressing with imidacloprid on the seedlings growth and protective enzyme activities of high-yielding summer maize].

A field plot experiment was conducted to study the effects of seed dressing with imidacloprid at the pesticide/seed mass ratios of 6:10000, 9:10000, and 12:10000 on the seedlings growth and protective enzyme activities of high-yielding summer maize. Seed dressing with an imidacloprid/seed mass ratio of 6:10000 increased the seedlings height, primary root length, secondary root number, aboveground fresh mass, and underground fresh mass by 2.81 cm, 2.31 cm, 1.71, 0.30 g, and 8.28 g, respectively, as compared to the control, and had better effect than the treatments 9:10000 and 12:10000. Comparing with the control, treatment 6:10000 increased the leaf- and root soluble protein content by 12.6% and 27.9% and catalase (CAT) activity by 2.5% and 11.5%, decreased malondialdehyde (MDA) content by 26.3% and 60.9%, improved superoxide dismutase (SOD) activity by 19.7% and 5.6%, enhanced glutathione-S-transferase (GSTs) activity by 11.5% and 13.2%, respectively, and increased the root peroxidase (POD) activity by 31.8%. Seed dressing with imidacloprid at the pesticide/seed mass ratio of 6:10000 could markedly improve maize seedlings protective enzyme activities, but had no promotion effect on leaf peroxidase (POD) activity.

[Yield loss model and yield loss mechanism of high-yielding summer maize infected by Physoderma maydis].

A total of 21 different disease-grading summer maize groups were formed by fixed-point natural infection of maize brown spot in the field, and mass loss estimation models of single ear mass and 100-grain mass were constructed by stepwise regression with DPS software. The mass loss estimation models of single ear and 100-grain were Y = -4.012 + 0.377X1 - 0.228X2 + 0.694X3 - 0.144X4 and Y = -4.536 + 0.173X1 + 0.188X2 + 0.248X3 - 0.034X4, respectively, where Y was yield loss rate, X1 was the disease index at flowering stage, X2 was the disease index at pollination stage, X3 was the disease index at filling stage, and X4 was the disease index at dough stage. The measured relationships between the disease indices at different growth stages and the mass loss for single ear and 100-grain coincided well with the modeling results. Maize brown spot directly affected the net photosynthetic rate of ear height leaf and the activities of RuBP carboxylase and PEP carboxylase. The higher the disease-grade, the lower the net photosynthetic rate and the activities of the two enzymes were.