Exploring the influence of invasive weed biochar on the sorption and dissipation dynamics of imazethapyr in sandy loam soil.

The management of invasive weeds on both arable and non-arable land is a vast challenge. Converting these invasive weeds into biochar and using them to control the fate of herbicides in soil could be an effective strategy within the concept of turning waste into a wealth product. In this study, the fate of imazethapyr (IMZ), a commonly used herbicide in various crops, was investigated by introducing such weeds as biochar, i.e., Parthenium hysterophorus (PB) and Lantana camara (LB) in sandy loam soil. In terms of kinetics, the pseudo-second order (PSO) model provided the best fit for both biochar-mixed soils. More IMZ was sorbed onto LB-mixed soil compared to PB-mixed soil. When compared to the control (no biochar), both PB and LB biochars (at concentrations of 0.2% and 0.5%) increased IMZ adsorption, although the extent of this effect varied depending on the dosage and type of biochar. The Freundlich adsorption isotherm provided a satisfactory explanation for IMZ adsorption in soil/soil mixed with biochar, with the adsorption process exhibiting high nonlinearity. The values of Gibb's free energy change (ΔG) were negative for both adsorption and desorption in soil/soil mixed with biochar, indicating that sorption was exothermic and spontaneous. Both types of biochar significantly affect IMZ dissipation, with higher degradation observed in LB-amended soil compared to PB-amended soil. Hence, the findings suggest that the preparation of biochar from invasive weeds and its utilization for managing the fate of herbicides can effectively reduce the residual toxicity of IMZ in treated agroecosystems in tropical and subtropical regions.

Transcriptomic and metabolomic analysis provides insight into imazethapyr toxicity to non-target plants.

Imazethapyr is a widely used imidazolinone herbicide worldwide, and its potential adverse effects on non-target plants have raised concerns. Understanding the mechanisms of imazethapyr phytotoxicity is crucial for its agro-ecological risk assessment. Here, the comprehensive molecular responses and metabolic alterations of Arabidopsis in response to imazethapyr were investigated. Our results showed that root exposure to imazethapyr inhibited shoot growth, reduced chlorophyll contents, induced photoinhibition and decreased photosynthetic activity. By non-target metabolomic analysis, we identified 75 metabolites that were significantly changed after imazethapyr exposure, and they are mainly enriched in carbohydrate, lipid and amino acid metabolism. Transcriptomic analysis confirmed that imazethapyr significantly downregulated the genes involved in photosynthetic electron transport and the carbon cycle. In detail, 48 genes in the photosynthetic lightreaction and 11 genes in Calvin cycle were downregulated. Additionally, the downregulation of genes related to electron transport in mitochondria provides strong evidence for imazethapyr inhibiting photosynthetic carbon fixation and cellular energy metabolism as one of mechanisms of toxicity. These results revealed the molecular and metabolic basis of imazethapyr toxicity on non-target plants, contributing to environmental risk assessment and mitigate negative impact of imazethapyr residues in agricultural soils.

Evaluating the imazethapyr herbicide mediated regulation of phenol and glutathione metabolism and antioxidant activity in lentil seedlings.

The imidazolinone group of herbicides generally work for controlling weeds by limiting the synthesis of the aceto-hydroxy-acid enzyme, which is linked to the biosynthesis of branched-chain amino acids in plant cells. The herbicide imazethapyr is from the class and the active ingredient of this herbicide is the same as other herbicides Contour, Hammer, Overtop, Passport, Pivot, Pursuit, Pursuit Plus, and Resolve. It is commonly used for controlling weeds in soybeans, alfalfa hay, corn, rice, peanuts, etc. Generally, the herbicide imazethapyr is safe and non-toxic for target crops and environmentally friendly when it is used at low concentration levels. Even though crops are extremely susceptible to herbicide treatment at the seedling stage, there have been no observations of its higher dose on lentils (Lens culinaris Medik.) at that stage. The current study reports the consequence of imazethapyr treatment on phenolic acid and flavonoid contents along with the antioxidant activity of the phenolic extract. Imazethapyr treatment significantly increased the activities of several antioxidant enzymes, including phenylalanine ammonia lyase (PAL), phenol oxidase (POD), glutathione reductase (GR), and glutathione-s-transferase (GST), in lentil seedlings at doses of 0 RFD, 0.5 RFD, 1 RFD, 1.25 RFD, 1.5 RFD, and 2 RFD. Application of imazethapyr resulted in the 3.2 to 26.31 and 4.57-27.85% increase in mean phenolic acid and flavonoid content, respectively, over control. However, the consequent fold increase in mean antioxidant activity under 2, 2- diphenylpicrylhdrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assay system was in the range of 1.17-1.85 and 1.47-2.03%. Mean PAL and POD activities increased by 1.63 to 3.66 and 1.71 to 3.35-fold, respectively, in agreement with the rise in phenolic compounds, indicating that these enzyme's activities were modulated in response to herbicide treatment. Following herbicide treatments, the mean thiol content also increased significantly in corroboration with the enhancement in GR activity in a dose-dependent approach. A similar increase in GST activity was also observed with increasing herbicide dose.

Exploration of the biodegradation pathway and enhanced removal of imazethapyr from soil by immobilized Bacillus marcorestinctum YN1.

Excessive or inappropriate applications of imazethapyr cause severe ecological deteriorations and health risks in human. A novel bacterial strain, i.e., Bacillus marcorestinctum YN1, was isolated to efficiently degrade imazethapyr, with the degradation pathways and intermediates predicted. Protein mass spectrometry analysis identified enzymes in strain YN1 potentially involved in imazethapyr biodegradation, including methylenetetrahydrofolate dehydrogenase, carbon-nitrogen family hydrolase, heme degrading monooxygenase, and cytochrome P450. The strain YN1 was further immobilized with biochar (BC600) prepared from mushroom waste (i.e., spent mushroom substrate) by pyrolysis at 600 °C to evaluate its degrading characteristics of imazethapyr. Scanning electron microscope observation showed that strain YN1 was adsorbed in the rich pore structure of BC600 and the adsorption efficiency reached the maximum level of 88.02% in 6 h. Both energy dispersive X-ray and Fourier transform infrared spectroscopy analyses showed that BC600 contained many elements and functional groups. The results of liquid chromatography showed that biochar-immobilized strain YN1 (IBC-YN1) improved the degradation rate of imazethapyr from 79.2% to 87.4%. The degradation rate of imazethapyr by IBC-YN1 could still reach 81.0% in the third recycle, while the bacterial survival rate was 67.73% after 180 d storage at 4 °C. The treatment of IBC-YN1 significantly shortened the half-life of imazethapyr in non-sterilized soil from 35.51 to 11.36 d, and the vegetative growth of imazethapyr sensitive crop plant (i.e., Cucumis sativus L.) was significantly increased in soil remediated, showing that the inhibition rate of root length and fresh weight were decreased by 12.45% and 38.49% respectively. This study exhanced our understanding of microbial catabolism of imazethapyr, and provided a potential in situ remediation strategy for improving the soil environment polluted by imazethapyr.

Imazethapyr disrupts plant phosphorus homeostasis and acquisition strategies.

The deficiency of essential mineral nutrients caused by xenobiotics often results in plant mortality or an inability to complete its life cycle. Imazethapyr, a widely utilized imidazolinone herbicide, has a long-lasting presence in the soil-plant system and can induce toxicity in non-target plants. However, the effects of imazethapyr on mineral nutrient homeostasis remain poorly comprehended. In this study, Arabidopsis seedlings exposed to concentrations of 4 and 10 µg/L imazethapyr showed noticeable reductions in shoot development and displayed a distinct dark purple color, which is commonly associated with phosphorus (P) deficiency in crops. Additionally, the total P content in both the shoots and roots of Arabidopsis significantly decreased following imazethapyr treatment when compared to the control groups. Through the complementary use of physiological and molecular analyses, we discovered that imazethapyr hinders the abundance and functionality of inorganic phosphorus (Pi) transporters and acid phosphatase. Furthermore, imazethapyr impairs the plant's Pi-deficiency adaptation strategies, such as inhibiting Pi transporter activities and impeding root hair development, which ultimately exacerbate P starvation. These results provide compelling evidence that residues of imazethapyr have the potential to disrupt plant P homeostasis and acquisition strategies. These findings offer valuable insights for risk assessment and highlight the need to reconsider the indiscriminate use of imazethapyr, particularly under specific scenarios such as nutrient deficiency.

The various effect of cow manure compost on the degradation of imazethapyr in different soil types.

Adding compost to soil is an effective strategy to promote the degradation of organic pollutants and reduce ecological risks. However, the effect of compost on the degradation of imazethapyr (IMET) in different soil types is not clear. To address this issue, a pot experiment was conducted, and high-throughput sequencing and mass spectrometry technology were used to identify the influence of cow manure compost on the degradation efficiency of IMET in black soil and saline-alkali soil and the role of key microorganisms. The results showed that adding compost to black soil increased the degradation rate of IMET by 12.58% and shortened the half-life by 53.37%, while in saline-alkali soil, the degradation rate of IMET decreased by 6.99% with no significant change in the half-life. High-throughput sequencing results showed that adding cow manure compost (mass ratio of 4%) significantly increased the abundance of bacterial families capable of degrading organic pollutants in black soil, but had an inhibitory effect on this bacterial community in saline-alkali soil. Redundancy analysis (RDA) results showed that total organic carbon (TOC), alkali-hydrolyzable nitrogen (AN), ammonia nitrogen (NH4+-N) and nitrate nitrogen (NO3--N) were the main factors driving microbial community variation. Mass spectrometry analysis indicated that IMET generated three metabolites during the degradation process. Sphingomonadaceae and Vicinamibacteraceae could accelerate the breaking of side-chain alkyl groups, while Chitinophagaceae could cause the rearrangement of the imidazole ring structure, gradually metabolizing IMET into small organic molecules. The application of appropriate cow manure compost can promote the development of IMET-degrading bacteria by adjusting the organic carbon and dissolved nitrogen content in black soil. In the future, the quantitative effects of organic fertilizer application on the IMET degradation process in different soil types should be further analyzed, and microbial isolation and purification should be used to enhance the ability of microorganisms to degrade herbicides.

Assessing the Stability of Herbicide-Tolerant Lentil Accessions (Lens culinaris Medik.) under Diverse Environments.

Assessing the adaptability and stability of herbicide-tolerant lentil accessions to two broad-spectrum post-emergence herbicides in multi-environment trials has become a must in a breeding program to improve its selection. The adaptability and stability of 42 herbicide-tolerant lentil accessions were investigated using five stability parameters under eight different environments. Significant Genotype-Environment (GE) interaction was found for days to flowering (DFLR), days to maturity (DMAT), and seed yield per plant (SY). The analyzed stability parameters such as Cultivar superiority, Finlay-Wilkinson, Shukla, Static Stability, and Wricke's Ecovalence ranked the tested accessions differently, confirming the importance of using a combination of stability parameters when evaluating the performance of a group of accessions. GGE biplot of the SY trait accounted for 60.79% of sums of squares of the GE interaction and showed that cool and high rainfall environments are ideal for testing the agronomic performance of tolerant accessions. The GGE biplot of SY showed that IG4605(19), IG195(6), and IG156635(12) were specifically adapted to one mega environment, whereas IG70056(38) was identified as a superior line having a high and stable yield. These lines should be included in lentil crossing programs to develop herbicide-tolerant cultivars adapted to diverse environments.

Effect of herbicide stress on synchronization of carbon and nitrogen metabolism in lentil (Lens culinaris Medik.).

Weed invasion causes significant yield losses in lentil. Imazethapyr (IM), a broad-spectrum herbicide inhibits the biosynthesis of branched chain amino acids necessary for plant growth. Plant growth depends upon translocation of photo-assimilates and their partitioning regulated by carbon and nitrogen metabolism. This study aimed to investigate the impact of imazethapyr spray on carbon and nitrogen metabolism in tolerant (LL1397 and LL1612) and susceptible (FLIP2004-7L and PL07) lentil genotypes during vegetative and reproductive development. Significantly higher activities of invertases and sucrose synthase (cleavage) in leaves and in podwall and seeds during early phase of development in tolerant genotypes were observed as compared to susceptible genotypes under herbicide stress that might be responsible for providing hexoses required for their growth. Activities of sucrose synthesizing enzymes, sucrose phosphate synthase and sucrose synthase (synthesis) increased significantly in podwalls and seeds of LL1397 and LL1612 genotypes during later phase of development towards maturity while the activities decreased in FLIP2004-7L and PL07 genotypes under herbicide stress. Activities of nitrate and nitrite reductase, glutamine 2-oxoglutarate aminotransferase, glutamine synthetase and glutamate dehydrogenase were increased in leaves, podwalls and seeds of LL1397 and LL1612 under herbicide stress. A proper synchronization of carbon and nitrogen metabolism in tolerant lentil genotypes during vegetative and reproductive phase might be one of the mechanisms for their recovery from herbicide stress. This first ever comprehensive information will provide a basis for future studies on the molecular mechanism of source sink relationship in lentil under herbicide stress and will be utilized in breeding programmes.

Impact of post-emergent imazethapyr on morpho-physiological and biochemical responses in lentil (Lens culinaris Medik.).

Yield reduction in lentil crop due to weed infestation is a key hindrance to its growth due to poor weed-crop competition. Imazethapyr (IM), a selective herbicide, target acetolactate synthase (ALS) which catalyzes the first reaction in biosynthesis of branched chain amino acids, required for plant growth and development. The objective of the present study was to investigate the impact of IM treatment on weeds, ALS enzyme activity, antioxidant capacity, osmolyte accumulation, growth and yield related parameters in lentil genotypes. Two IM tolerant (LL1397 and LL1612) and two susceptible (FLIP2004-7L and PL07) lentil genotypes were cultivated under weed free, weedy check and IM treatments. Weed control efficiency reached its peak at 21 days after spray (DAS). Imazethapyr treatment decreased chlorophyll and carotenoid content up to 28 DAS with higher reduction in susceptible genotypes. FLIP2004-7L and PL07 had reduced plant height and lower number of pods under IM treatment which resulted in decreased seed yield. Higher ALS activity in LL1397 and LL1612 at 21 DAS, higher antioxidant capacity and glycine betaine content both at 21 and 28 DAS and lower decrease in relative leaf water content might be mediating herbicide tolerance in these genotypes that led to higher seed yield. The identified IM tolerance mechanism can be used to impart herbicide resistance in lentil. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01244-x.

Multiple resistance to ALS-inhibiting and PPO-inhibiting herbicides in Chenopodium album L. from China.

Common lambsquarters (Chenopodium album L.) is a broadleaf weed that can severely damage soybean fields. Two C. album populations (1744 and 1731) suspected resistant to imazethapyr were investigated for resistance levels to imazethapyr, thifensulfuron-methyl, and fomesafen and their resistance mechanisms were investigated. Whole-plant dose-response assays revealed that, compared to the susceptible (S) population, the 1744 population was 16.5-fold resistant to imazethapyr, slightly resistant to thifensulfuron-methyl (resistance index [R/S], <3). The 1731 population was 18.8-fold resistant to imazethapyr, 2.9-fold resistant to thifensulfuron-methyl, and 5.1-fold resistant to fomesafen. In vitro acetolactate synthase (ALS) assays showed 17.1-fold and 19.3-fold resistance levels of 1744 and 1731 populations to imazethapyr respectively. ALS gene sequence analysis identified Ala122Thr amino acid substitution in the 1744 population and Ser653Thr amino acid substitution in the 1731 population. No mutations of the protoporphyrinogen oxidase (PPO) gene were detected. However, pre-treatment with malathion reversed fomesafen resistance, suggesting nontarget-site resistance mechanisms likely play a role in the 1731 population.

A novel naturally Phe206Tyr mutation confers tolerance to ALS-inhibiting herbicides in Alopecurus myosuroides.

Herbicide-resistant weeds pose a serious threat to world food production. The rapid and widespread development of target-site based resistance limits the application of herbicides. Alopecurus myosuroides Huds. (blackgrass) has spread rapidly in winter wheat regions in China, and the field recommended dose of ALS herbicides no longer controls blackgrass populations in recent years. A highly resistant population TW18(R) was collected in 2018 from Shandong Province. Dose-response assays showed that the TW18 was resistant to mesosulfuron-methyl, flucarbazone-sodium, and imazethapyr, with resistance index values of 5.96, 6.1, and 4.09, respectively. DNA sequencing of the TW18 population revealed a Phe206Tyr (F206Y) mutation in the ALS, which was not yet reported. Blackgrass ALS gene with the F206Y mutation (R gene) was expressed in Arabidopsis and rice. Transgenic studies have shown that both Arabidopsis and rice expressing this R gene have resistance to imazethapyr. However, it did not confer resistance to tribenuron methyl and florasulam in transgenic Arabidopsis. This study showed that the F206Y substitution caused herbicide resistance in blackgrass. To our knowledge, this is the first-reported F206Y mutation of a weed species in the natural environment. Transgenic plants showed this functional site could be utilized to generate imazethapyr-resistant rice to control herbicide-resistant weed damage.

Methylglyoxal detoxification pathway - Explored first time for imazethapyr tolerance in lentil (Lens culinaris L.).

Lentil is an important pulses crop but it's short stature and slow growth rate make it vulnerable to weed competition, limiting crop productivity. There is need to identify herbicide tolerant genotypes and their tolerance mechanism. The present investigation was conducted to understand the effect of imazethapyr (IM) treatment on accumulation of methylglyoxal (MG) and its detoxification mechanism in IM-tolerant (LL1397 and LL1612) susceptible (FLIP2004-7L and PL07) genotypes sown under control (weed free), weedy check (weeds were growing with crop) and sprayed with imazethapyr. The enzymes of glyoxalase pathway (glyoxalase I, II and III) and non glyoxalase pathway (methylglyoxal reductase), lactate dehydrogenase (LDH), glutathione content, gamma-glutamyl-cysteine synthetase (γ-GCS) were estimated in lentil genotypes at different days after spray. Higher activities of glyoxalase I, II and III and MGR along with the increased glutathione content (GSH) content in LL1397 and LL1612 under IM treatment as compared to FLIP2004-7L and PL07 might be responsible for lowering MG accumulation and increasing lactate content, which is end product of these pathways. Enhanced LDH activity in LL1397 and LL1612 might be responsible for energy production via TCA cycle that might be responsible for growth and recovery of tolerant genotypes after IM treatment. Higher γ-GCS activity in tolerant genotypes led to increased glutathione content required for glyoxalase pathway. However, decreased activities of glyoxalase enzymes and MGR in susceptible genotypes result in MG accumulation which limit plant growth. This is the first ever study elucidating the role of MG detoxification pathway conferring IM tolerance in lentil.

Tribenuron-methyl-resistant Descurainia sophia L. exhibits negative cross-resistance to imazethapyr conferred by a Pro197Ser mutation in acetolactate synthase and reduced metabolism.

BACKGROUND: Descurainia sophia L. is one of the most notorious weeds infesting winter wheat in China. Mutations at Pro197 in acetolactate synthase (ALS) results in resistance of D. sophia to tribenuron-methyl and cross-resistance to many ALS inhibitors. Negative cross-resistance to imazethapyr was observed in tribenuron-methyl-resistant (TR) D. sophia with the Pro197Ser mutation in a previous study. In the present research, another TR D. sophia with the Pro197Ser mutation was obtained. To explore the mechanisms of negative cross-resistance, the ALS sensitivity, the absorption and metabolism of imazethapyr in tribenuron-methyl-susceptible (TS) and TR D. sophia were studied. RESULTS: The TR D. sophia population with the Pro197Ser mutation (pHB23) displayed negative cross-resistance to imazethapyr and no cross-resistance to imazamox and imazapic. In contrast, TR D. sophia populations with other Pro197 mutations had no or low resistance to imazethapyr. The ALS in the pHB23 population was more susceptible to imazethapyr than that in the TS population. There was no difference in the absorption of imazethapyr, imazamox, and imazapic between TS and pHB23 plants. However, the metabolism of imazethapyr in TS D. sophia was faster than that in pHB23 plants up to 1 week after treatment. There was no significant difference in the metabolism of imazamox and imazapic between TS and pHB23 plants. CONCLUSION: The TR D. sophia population with the Pro197Ser mutation exhibited negative cross-resistance to imazethapyr, which was likely due to reduced metabolism and increased sensitivity of ALS to imazethapyr. © 2021 Society of Chemical Industry.

Bioavailability and toxicity of imazethapyr in maize plant estimated by four chemical extraction techniques in different soils.

The bioavailability and toxicity of herbicides on the crop depend on its uptake efficiency from the soil, and thus the assessment of the bioavailable fraction of herbicides in soil is a crucial work. In this study, we investigated the uptake concentration and toxicity of imazethapyr in maize plant using four chemical measurements, including the extraction of in situ pore water (CIPW), ex situ pore water (CEPW), organic solvent (Csoil) and passive sampling (Cfree) in five soils. The results obtained that the CIPW in a specific soil had the most significant correlation with the uptake concentration of imazethapyr in maize plant (R2 = 0.8851-0.9708), followed by CEPW (R2 = 0.8911-0.9565) and Cfree (R2 = 0.7881-0.9673). However, Cfree showed a higher correlation when considering all five soils, and thus Cfree can describe the bioavailability beyond the types of soil. Additionally, the median inhibition concentrations (IC50) of imazethapyr to maize plant ranged from 5.0 to 6.9 mg/kg in five soils, and the CIPW, CEPW and Cfree had better relationships with the IC50 (R2 > 0.8681) than the Csoil (R2 = 0.6782). The effects of soil properties on the phytotoxicity of imazethapyr, including pH, organic matter content, cation exchange capacity and clay content, were studied, and the soil pH was shown to be a main factor. This study demonstrated that the freely dissolved fraction and soil pore water concentration of imazethapyr in soil can be used to evaluate its bioavailability and toxicity to maize.

De novo genome assembly of a foxtail millet cultivar Huagu11 uncovered the genetic difference to the cultivar Yugu1, and the genetic mechanism of imazethapyr tolerance.

BACKGROUND: Setaria italica is the second-most widely planted species of millets in the world and an important model grain crop for the research of C4 photosynthesis and abiotic stress tolerance. Through three genomes assembly and annotation efforts, all genomes were based on next generation sequencing technology, which limited the genome continuity. RESULTS: Here we report a high-quality whole-genome of new cultivar Huagu11, using single-molecule real-time sequencing and High-throughput chromosome conformation capture (Hi-C) mapping technologies. The total assembly size of the Huagu11 genome was 408.37 Mb with a scaffold N50 size of 45.89 Mb. Compared with the other three reported millet genomes based on the next generation sequencing technology, the Huagu11 genome had the highest genomic continuity. Intraspecies comparison showed about 94.97 and 94.66% of the Yugu1 and Huagu11 genomes, respectively, were able to be aligned as one-to-one blocks with four chromosome inversion. The Huagu11 genome contained approximately 19.43 Mb Presence/absence Variation (PAV) with 627 protein-coding transcripts, while Yugu1 genomes had 20.53 Mb PAV sequences encoding 737 proteins. Overall, 969,596 Single-nucleotide polymorphism (SNPs) and 156,282 insertion-deletion (InDels) were identified between these two genomes. The genome comparison between Huagu11 and Yugu1 should reflect the genetic identity and variation between the cultivars of foxtail millet to a certain extent. The Ser-626-Aln substitution in acetohydroxy acid synthase (AHAS) was found to be relative to the imazethapyr tolerance in Huagu11. CONCLUSIONS: A new improved high-quality reference genome sequence of Setaria italica was assembled, and intraspecies genome comparison determined the genetic identity and variation between the cultivars of foxtail millet. Based on the genome sequence, it was inferred that the Ser-626-Aln substitution in AHAS was responsible for the imazethapyr tolerance in Huagu11. The new improved reference genome of Setaria italica will promote the genic and genomic studies of this species and be beneficial for cultivar improvement.

Insights into the Role of Transcriptional Gene Silencing in Response to Herbicide-Treatments in Arabidopsis thaliana.

Herbicide resistance is broadly recognized as the adaptive evolution of weed populations to the intense selection pressure imposed by the herbicide applications. Here, we tested whether transcriptional gene silencing (TGS) and RNA-directed DNA Methylation (RdDM) pathways modulate resistance to commonly applied herbicides. Using Arabidopsis thaliana wild-type plants exposed to sublethal doses of glyphosate, imazethapyr, and 2,4-D, we found a partial loss of TGS and increased susceptibility to herbicides in six out of 11 tested TGS/RdDM mutants. Mutation in REPRESSOR OF SILENCING 1 (ROS1), that plays an important role in DNA demethylation, leading to strongly increased susceptibility to all applied herbicides, and imazethapyr in particular. Transcriptomic analysis of the imazethapyr-treated wild type and ros1 plants revealed a relation of the herbicide upregulated genes to chemical stimulus, secondary metabolism, stress condition, flavonoid biosynthesis, and epigenetic processes. Hypersensitivity to imazethapyr of the flavonoid biosynthesis component TRANSPARENT TESTA 4 (TT4) mutant plants strongly suggests that ROS1-dependent accumulation of flavonoids is an important mechanism for herbicide stress response in A. thaliana. In summary, our study shows that herbicide treatment affects transcriptional gene silencing pathways and that misregulation of these pathways makes Arabidopsis plants more sensitive to herbicide treatment.

Enantioselectivity effects of imazethapyr enantiomers to metabolic responses in mice.

Imazethapyr (IMZT) is a typical chiral pesticide with two enantiomers with the R-IMZT having the main herbicidal activity. However, the enantioselectivity of the effects of IMZT enantiomers on human and animals is still unclear. In this study, a nuclear magnetic resonance (NMR)-based metabolomics method and determination of oxidative stress were used to evaluate the enantioselectivity of IMZT enantiomers in mice. The results showed that the R-IMZT caused larger disturbances of endogenous metabolites and the S-IMZT had stronger interferences to oxidation defense system. The significantly perturbed metabolic pathways in mice exposed to the R-enantiomer were the valine, leucine and isoleucine biosynthesis pathway as well as the phenylalanine, tyrosine and tryptophan biosynthesis pathway. However, exposure of mice to the S-enantiomer did not significantly affect the metabolic pathways, but exposure led to an increase of catalase (CAT) activity and an increase in malondialdehyde (MDA) content in the liver. These results indicate that we need to conduct a more comprehensive assessment of the health risks of pesticide monomers in the future. In a word, these results provide more evidence for assessing the differences in health risks of IMZT enantiomers to mammals as well as provide more references for the promotion and use of pesticide monomers in the future.

Phytostimulation of lowland soil contaminated with imidazolinone herbicides.

The phytostimulation is a phytoremediation technique that can be used to remediate area contaminated with herbicides. It is necessary to select plants with high capacity to stimulate soil microbial activity. The present work aimed at evaluating seven plant species regarding their ability to phytostimulate soil and enhance the degradation of the herbicides imazethapyr, imazapic and imazapyr in a lowland soil. An Alfisol Albaqualf was cultivated with the following species, Canavalia ensiformis, Glycine max, Oryza sativa cultivar PUITÁ INTA CL, Lolium multiflorum, Vicia sativa and consortium Lotus corniculatus + Trifolium repens. The rhizosphere of these plants and non-rhizospheric (uncultivated soil) as a control were contaminated in laboratory with analytical standart of the three herbicides, at rates of 0, 150, 300 and 750 g a.i. ha-1, in separate assays. Biodegradation was estimated by quantifying C-CO2 production and through analysis of herbicides residues in soil using liquid chromatography. Results show that biodegradation of herbicides imazethapyr, imazapic and imazapyr was higher in vegetated soil than in unvegetated soil. The leguminous species Canavalia ensiformis, Glycine max, Vicia sativa and consortium of Lotus corniculatus + Trifolium repens showed a great capacity to promote soil microbial, resulting in average biodegradation rates of 91, 92 and 93% for herbicides imazethapyr, imazapic and imazapyr in soil, respectively.

An imazethapyr-based herbicide formulation induces genotoxic, biochemical, and individual organizational effects in Leptodactylus latinasus tadpoles (Anura: Leptodactylidae).

Genotoxic, biochemical, and individual organizational effects on Leptodactylus latinasus tadpoles were evaluated after exposure to an imazethapyr (IMZT)-based commercial herbicide formulation, Pivot® H (10.59% IMZT). A determination of the value of the lethal concentration (LC50) was determined as a toxicological endpoint. Alterations in animal behavior and morphological abnormalities as well as cholinesterase (ChE), catalase (CAT), and glutathione S-transferase (GST) activities were employed as individual sublethal endpoints. Micronuclei frequencies (MNs), binucleated cells (BNs), blebbed nuclei (BLs), lobed nuclei (LBs), notched nuclei (NTs), erythroplastids (EPs), and evaluation of DNA strand breaks were employed as genotoxic endpoints. All biomarkers were evaluated after 48 and 96 h of exposure to concentrations of IMZT within 0.07-4.89 mg/L. LC5096h values of 1.01 and 0.29 mg/L IMZT were obtained for Gosner stages 25 and 36, respectively. Irregular swimming, diamond body shape, and decreased frequency of keratodonts were detected at both sampling times. Results showed that IMZT increased GST activity and MN frequency at 48 and 96 h of exposure. Other nuclear abnormalities were also observed in the circulating erythrocytes of tadpoles, i.e., NT and BL values after 48 h, and LN, BL, and EP values after 96 h. Finally, results showed that IMZT within 0.07-0.22 mg/L increased the genetic damage index in tadpoles exposed for both exposure times (48 and 96 h). This study is the first to report the sublethal biochemical effects of IMZT in anurans and is also the first report using L. latinasus tadpoles as a bioindicator for ecotoxicological studies.

Comparative molecular and metabolic responses of wheat seedlings (Triticum aestivum L.) to the imazethapyr enantiomers S-IM and R-IM.

The enantioselective effects of imazethapyr (IM) enantiomers on wheat seedlings in a hydroponic medium were studied. R-IM at 0.05mg/L exerted a stronger inhibitory effect on shoot weight and root weight than 0.05mg/L S-IM, suggesting that R-IM more severely inhibited growth. Oxidative damage, based on the anthocyanin content, malondialdehyde (MDA) content, antioxidant enzyme activities and transcript levels of antioxidant enzyme genes, were studied together with the cellular ultrastructure of wheat leaves. The anthocyanin and MDA contents in the R-IM treatment group were significantly increased compared with those in the control group, but no significant changes were observed in the S-IM treatment group. The antioxidant enzyme activities of CAT and SOD were inhibited by 0.32- and 0.73-fold, respectively, in the 14day R-IM treatment group compared to those in the control. However, the transcript levels of antioxidant enzyme genes, including CuZnSOD, POD and CAT, were downregulated in the 14day R-IM exposure group, but those of DHAR were not. The number and size of starch granules increased and chloroplast swelling was observed in wheat leaf cells after R-IM exposure, which showed that photosynthetic functions were potentially disturbed. These results directly or indirectly imply that R-IM exposure causes more oxidative stress and exerts a stronger negative effect on wheat than S-IM. A metabolomics approach revealed that the tricarboxylic acid cycle was heavily suppressed by R-IM treatment. Some amino acids (proline, threonine, lysine, valine) were increased by only the R-IM treatment, indicating the activation of antioxidant pathways. The decrease in a series of fatty acids implied that the cell membrane composition changed in response to R-IM. These results provide a deeper understanding of the enantioselective effects of IM enantiomers on the molecular and metabolic responses in wheat seedlings.