•Thifluzamide increased liver glycogen levels in zebrafish.•Thifluzamide reduced blood glucose levels in zebrafish.•Aerobic glycolysis in liver was inhibited by thifluzamide.•Anaerobic glycolysis in liver was inhibited by thifluzamide.•Pentose phosphate pathway in liver was stimulated by thifluzamide.Thifluzamide exerts toxic effects to zebrafish and causes liver mitochondrial damage. To better understand the further mechanism, adult zebrafish were exposed to a range of thifluzamide concentrations (0, 0.019, 0.19, and 1.90 mg/L) for 28 days. In response to 1.90 mg/L exposure, liver glycogen significantly increased and blood glucose decreased. The expression of genes related to glycometabolism showed corresponding changes. Genes related to mtDNA replication and transcription and genes participating in mitochondrial complexes showed altered expression, which might lead to the inhibition of the tricarboxylic acid cycle (TCA). Additionally, the activity of glucose-6-phosphate dehydrogenase (G6PDH) was markedly increased at 1.90 mg/L, which might result in the activation of the pentose phosphate pathway. Moreover, the activity of lactate dehydrogenase (LDH) was significantly reduced at 1.90 mg/L, which might indicate that anaerobic glycolysis was inhibited. This study suggests that the altered gene expression and enzyme activities might be responsible for changes in glycometabolism, as evidenced by the altered expression of glycometabolism-related genes, the increased amount of glycogen in the liver and the decreased blood glucose levels. Overall, thifluzamide caused dysfunctional glycometabolism and led to events that might contribute to various thifluzamide-induced abnormalities in zebrafish.

ZmGST34 is a maize Tau class GST gene and was found to be differently expressed between two maize cultivars differing in tolerance to herbicide metolachlor. To explore the possible role of ZmGST34 in maize development, the expression pattern and substrate specificity of ZmGST34 were characterized by quantitative RT-PCR and heterologous expression system, respectively. The results indicated that the expression level of ZmGST34 was increased ∼2–5-fold per day during the second-leaf stage of maize seedling. Chloroacetanilide herbicides or phytohormone treatments had no influence on the expression level of ZmGST34, suggesting that ZmGST34 is a constitutively expressed gene in maize seedling. Heterologous expression in Escherichia coli and in Arabidopsis thaliana proved that ZmGST34 can metabolize most chloroacetanilide herbicides and increase tolerance to these herbicides in transgenic Arabidopsis thaliana. The constitutive expression pattern and broad substrate activity of ZmGST34 suggested that this gene may play an important role in maize development in addition to the detoxification of pesticides.Keywords: chloroacetanilide herbicides; constitutive expression; expression pattern; heterologous expression; substrate specificity; ZmGST34;

Glutathione S-transferases (GSTs) play important roles in herbicide tolerance. However, studies on GST function in herbicide tolerance among plant tissues are still lacking. To explore the mechanism of metolachlor tolerance difference between maize shoots and roots, the effects of metolachlor on growth, GST activity, and the expression of the entire GST gene family were investigated. It was found that this differential tolerance to metolachlor was correlated with contrasting GST activity between the two tissues and can be eliminated by a GST inhibitor. An in vitro metolachlor–glutathione conjugation assay confirmed that the transformation of metolachlor is 2-fold faster in roots than in shoots. The expression analysis of the GST gene family revealed that most GST genes are expressed much higher in roots than shoots, both in control and in metolachlor-treated plants. Taken together, higher level expression of most GST genes, leading to higher GST activity and faster herbicide transformation, appears to be responsible for the higher tolerance to metolachlor of maize roots than shoots.Keywords: glutathione S-transferases; herbicide tolerance; maize; metolachlor; shoots and roots;

•Variations in metolachlor tolerance were identified among 10 maize cultivars in China.•Metabolism of metolachlor in leaves of Nongda86 was more rapid than in Zhengdan958 and could be inhibited by ETA.•GST gene expression and activity were higher in Nongda86 than Zhengdan958 following metolachlor treatment.Glutathione S-transferases (GSTs) have been widely studied in relation to their role in herbicide tolerance and detoxification. However, a detailed characterization of GSTs from herbicide tolerant and sensitive maize cultivars is still lacking. In this study, we determined the mechanism of differential tolerance between two maize cultivars which had 4-fold difference tolerance to metolachlor. The metabolism rate of metolachlor was more rapid in the tolerant cultivar (Zea mays L. cv Nongda86) than the susceptible one (Zea mays L. cv Zhengda958). Addition of the GST inhibitor ethacrynic acid reduced the metabolism of metolachlor indicating the involvement of GSTs in the differential detoxification of metolachlor. The expression profiles of 32 GST isozymes were measured using quantitative RT-PCR. The results showed the expression of GST genes were slightly up-regulated in Nongda86, but severely inhibited in Zhengdan958 24 h after metolachlor treatment. The genes GSTI, GSTIII, GSTIV, GST5, GST6 and GST7, which can detoxify chloroacetanilide herbicides, were all expressed higher in Nongda86 compared to Zhendgan958. The result of GST activity was consistent with the gene expression profiles. Collectively, higher-level expression of GST genes, leading to higher GST activity and faster herbicide detoxification, appears to be responsible for the difference in tolerance to metolachlor in two maize cultivars.Download high-res image (71KB)Download full-size image

•Exposure difenoconazole affected female zebrafish gonad index and hepatic index.•The levels of hormones in GH/IGF axis were altered by difenoconazole.•Difenoconazole disrupted the GH/IGF system of zebrafish in a sex-specific manner.•The content of difenoconazole, difenoconazole alcohol was higher in females than in males.Difenoconazole, as one of the most widely used triazole fungicides, is applied to protect crops, fruits, and vegetables. It has been reported that difenoconazole can enter the environment and impair aquatic organisms, but whether difenoconazole can disrupt the growth hormone (GH) balance in adult zebrafish (Danio rerio) is still unclear. In this study, adult female and male zebrafish were exposed to difenoconazole (0, 5, 50, and 500 µg/L) for seven days. The results revealed that the bioaccumulation of difenoconazole and its primary metabolite difenoconazole alcohol in females were both larger than that in males. In females, the growth of the liver and ovary were inhibited, which may be due to the decreased transcription of the key genes igf1a, igf2a, and igf2b in both organs. Male fish growth was promoted in response to the increased expression of genes relevant to the GH/insulin-like growth factor axis (GH/IGF) axis in the brain, liver, and testis as well as increased GH levels. It was found that difenoconazole interfered with the growth endocrine system and sex-specifically altered the expression of GH/IGF axis related genes in adult zebrafish after a short-term exposure.Download high-res image (186KB)Download full-size image

•Diamide insecticides affected survival, reproduction and growth of D. magna.•Diamide insecticides induced severe developmental abnormalities in D. magna embryos.•Diamide insecticides provoked oxidative stress in D. magna.•Even low levels of diamide insecticides pose ecological risks to aquatic ecosystems.The diamides have become one of the most promising new classes of insecticides. In this study, we evaluated the toxicity of three diamides (chlorantraniliprole, cyantraniliprole and flubendiamide) to Daphnia magna. The acute toxicity test showed that the 48-h EC50 of chlorantraniliprole, cyantraniliprole and flubendiamide were 8.5, 23.9 and 63.5 μg/L, respectively. Biochemical measurements revealed a significant increase in reactive oxygen species (ROS) in D. magna after acute exposure to the three diamides. A significant decrease in activities of antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) was observed, which was consistent with the down-regulated transcription of antioxidant genes sod and gpx. Catalase (CAT) activity exhibited a significant increase while the related gene cat showed no obvious change in daphnids acutely exposed to the three diamides. The chronic test revealed that the three diamides could cause lethal and sub-lethal effects on daphnids within constricted range of concentrations at μg/L level. The 21-d EC50 of chlorantraniliprole, cyantraniliprole and flubendiamide for mobility were 5.0, 13.6 and 36.8 μg/L, respectively. The chronic LOEC of chlorantraniliprole, cyantraniliprole and flubendiamide based on survival, growth and reproduction of D. magna were 4.05, 10.24 and 19.36 μg/L, respectively. Moreover, these three diamides can induce severe developmental abnormalities in D. magna embryos including underdeveloped second antennae, curved tail spine and abnormal body region after acute exposure and the 48-h EC50 were 6.2, 14.1 and 30.8 μg/L for chlorantraniliprole, cyantraniliprole and flubendiamide respectively. Our findings indicate that even low levels of diamides can pose ecological risks to aquatic ecosystems.

In this study, we applied various developmental stages of zebrafish to address the potential environmental risk and aquatic toxicity of bromothalonil and flutolanil. This results demonstrated that the acute toxicity of bromothalonil to the three phases of zebrafish were 4.34 (embryo) < 3.27 (12 h old larvae) < 2.52 mg/L (adult fish) and that of flutolanil were 5.47 (embryo) < 4.09 (72 h old larvae) < 3.91 (12 h old larvae) < 2.70 mg/L (adult). Sublethal effects induced by both bromothalonil and flutolanil on zebrafish embryos were noted, including growth inhibition, abnormal spontaneous movement, slower heart rate, complete hatching failure, and morphological deformities. In addition, both bromothalonil and flutolanil could cause notochord deformation and short body length of larvae. This study provides a foundation for future investigation into the mechanism of bromothalonil and flutolanil toxicity in zebrafish.

Imidacloprid is a well-known pesticide and it is timely to evaluate its toxicity to earthworms (Eisenia fetida). In the present study, the effect of imidacloprid on reproduction, growth, acetylcholinesterase (AChE) and DNA damage in earthworms was assessed using an artificial soil medium. The median lethal concentration (LC50) and the median number of hatched cocoons (EC50) of imidacloprid to earthworms was 3.05 and 0.92 mg/kg respectively, the lowest observed effect concentration of imidacloprid about hatchability, growth, AChE activity and DNA damage was 0.02, 0.5, 0.1 and 0.5 mg/kg, respectively.

The toxic effects of buprofezin on Daphnia magna after both chronic and acute exposures were evaluated according to OECD guidelines. A 48-h acute exposure of buprofezin resulted in daphnid immobility at an EC50 of 0.44 mg/L. In a 14 days chronic exposure of buprofezin (0, 0.025, 0.05, 0.10 and 0.15 mg/L), the development and reproduction of daphnids were all significantly affected and the body length was more sensitive than other observed parameters. However, the adverse effects of buprofezin on parental daphnids can be passed on to their offspring and cannot be recovered in a short time.

The oriental tobacco budworm, Helicoverpa assulta, is one of the most destructive pests for numerous commercial crops, and these organisms are responsible for enormous economic losses in Chinese agriculture. Insect larvae often feed within host plant fruits, providing protection from many currently used insecticides and making field control of H. assulta very difficult. Owing to its novel mode of action, high insecticidal activity, and low mammalian toxicity, the nonsystemic insecticide indoxacarb has been considered a promising alternative for the control of lepidopterous pests of agricultural significance. Indoxacarb evidences an elevated insecticidal activity against H. assulta. After 13 generations of selection with indoxacarb and bifenthrin insecticides under laboratory conditions, the LC50 of these compounds for H. assulta increased by 4.19-fold and 10.67-fold, respectively. The synergists diethyl maleate (DEM) and triphenyl phosphate (TPP) increased indoxacarb toxicity by 2.76-fold and 4.10-fold in resistant strains and, comparatively, 1.58-fold and 1.75-fold in susceptible strains, suggesting that carboxylesterase (CarE) and glutathione-S-transferases (GSTs) may be involved in the development of indoxacarb resistance in H. assulta. Activity and kinetic parameters observed in detoxification enzymes further demonstrated that the enhanced activity of CarE and GSTs may be critical in development of indoxacarb resistance in H. assulta. The data provides a foundation for further study of the indoxacarb resistance mechanism observed in H. assulta and the rational use of indoxacarb as a rotation insecticide with other insecticide classes for the control of H. assulta.Graphical abstractDownload full-size imageHighlights► The insecticidal activity of bifenthrin and indoxacarb against Helicoverpa assulta was higher than other test insecticides. ► H. assulta developed resistance to bifenthrin more readily than it developed resistance to indoxacarb. ► The resistance mechanism to indoxacarb in H. assulta was probably related to the enhanced activity of CarE and GSTs. ► Indoxacarb may be an ideal insecticide for the control of H. assulta.