Major Detoxification Enzymes Research Articles

KLF15-Cyp3a11 Axis Regulates Rifampicin-Induced Liver Injury.

Rifampicin (RFP) has demonstrated potent antibacterial effects in the treatment of pulmonary tuberculosis. However, the serious adverse effects on the liver intensively limit the clinical usage of the drug. Deacetylation greatly reduces the toxicity of RFP but also retains its curative activity. Here, we found that Krüppel-like factor 15 (KLF15) repressed the expression of the major RFP detoxification enzyme Cyp3a11 in mice via both direct and indirect mechanisms. Knockout of hepatocyte KLF15 induced the expression of Cyp3a11 and robustly attenuated the hepatotoxicity of RFP in mice. In contrast, overexpression of hepatic KLF15 exacerbated RFP-induced liver injury as well as mortality. More importantly, the suppression of hepatic KLF15 expression strikingly restored liver functions in mice even after being pretreated with overdosed RFP. Therefore, this study identified the KLF15-Cyp3a11 axis as a novel regulatory pathway that may play an essential role in the detoxification of RFP and associated liver injury. SIGNIFICANCE STATEMENT: Rifampicin has demonstrated antibacterial effects in the treatment of pulmonary tuberculosis. However, the serious adverse effects on the liver limit the clinical usage of the drug. Permanent depletion and transient inhibition of hepatic KLF15 expression significantly induced the expression of Cyp3a11 and robustly attenuated mouse hepatotoxicity induced by RFP. Overall, our studies show the KLF15-Cyp3a11 axis was identified as a novel regulatory pathway that may play an essential role in the detoxification of RFP and associated liver injury.

Functional analysis of UDP-glycosyltransferase genes conferring indoxacarb resistance in Spodoptera litura

UDP-glycosyltransferase (UGT) is the major detoxification enzymes of phase II involved in xenobiotics metabolism, which potentially mediates the formation of insect resistance. Previous transcriptome sequencing studies have found that several UGT genes were upregulated in indoxacarb resistant strains of Spodoptera litura, but whether these UGT genes were involved in indoxacarb resistance and their functions in resistance were unclear. In this study, the UGTs inhibitor, 5-nitrouracil, enhanced the toxicity of indoxacarb against S. litura, preliminarily suggesting that UGTs were participated in indoxacarb resistance. Two UGT genes, UGT33J17 and UGT41D10 were upregulated in the resistant strains and could be induced by indoxacarb. Alignment of UGT protein sequences revealed two conserved donor-binding regions with several key residues that interact with catalytic sites and sugar donors. Further molecular modeling and docking analysis indicated that two UGT proteins were able to stably bind indoxacarb and N-decarbomethoxylated metabolite (DCJW). Furthermore, knockdown of UGT33J17 and UGT41D10 decreased viability of Spli-221 cells and enhanced susceptibility of larvae to indoxacarb. Transgenic overexpression of these genes reduced the toxicity of indoxacarb in Drosophila melanogaster. This work revealed that upregulation of UGT genes significantly contributes to indoxacarb resistance in S. litura, and is of great significance for the development of integrated and sustainable management strategies for resistant pests in the field.

Decreased cuticular penetration minimizes the impact of the pyrethroid insecticide λ-cyhalothrin on the insect predator Eocanthecona furcellata

The use of broad-spectrum pesticides may reduce the biological control efficacy of predatory arthropods. Hence, the risks of pesticides to predators need to be evaluated. Here, we assessed the effects of a broad spectrum pyrethroid λ-cyhalothrin on a polyphagous predatory insect Eocanthecona furcellata via contact exposure route. The recommended application rate of λ-cyhalothrin was lower than the LR50 and HQ (in-field) was equal to 0.57, indicating the risk of λ-cyhalothrin to E. furcellata was low. Dried λ-cyhalothrin residue had no effect on the mortality, body weight, protein content of cuticle, or activities of major detoxification enzymes in E. furcellata. Residual of λ-cyhalothrin was only detected in the cuticle and legs of E. furcellata with a decreasing trend as time went by and no λ-cyhalothrin was detected inside the body. Additionally, a comparative transcriptome analysis was conducted to study global changes in gene expression in E. furcellata at different time points following exposure to λ-cyhalothrin-contaminated environment. A total of 57,839 unigenes with an average length of 1044 bp and an N50 of 1820 bp were obtained. In total, 118 and 109 differentially expressed genes (DEGs) at 12 h, and 60 h were identified between two groups. The DEGs were largely enriched in functional categories related to the structural constituent of cuticle. Accordingly, multiple cuticle protein-coding genes were up-regulated at 12 h after pesticide exposure. The present study stressed the importance of evaluating the compatibility between a specific pesticide (λ-cyhalothrin) and E. furcellata via simulating the releasing predators after insecticide application. The data could help optimize the pesticide use, optimizing the ecological services of E. furcellata as a BCA, and expanding its use into more areas of agriculture.

Pretreatment of mosquito larvae with ultraviolet-B and nitropolycyclic aromatic hydrocarbons induces increased sensitivity to permethrin toxicity

Nitropolycyclic aromatic hydrocarbons (Nitro-PAH) are highly toxic PHA derivatives. Nitro-PHAs are emitted by carbonaceous materials and PHA post-emission transformation, which causes water and environmental pollution and also exists as carcinogenic and immunotoxic agents. UV light has been shown to cause DNA damage and improves the covalent binding of PAH to DNA significantly. Mosquito breeding grounds are pools of water that can be large open zones or encased ponds with varying levels of sunlight exposure. This research was performed to assess the combined effects of UV-B exposure and Nitro-PAH on the physiological function of Culex quinquefasciatus larvae. To assess the impact of UV-B irradiation and Nitro-PAH exposure on mosquito vectors, parameters were examined: (1) Nitro-PAH availability and its impact on cell fatalities; (2) the detoxifying abilities of cytochrome P450, glutathione-S-transferase, and esterase; (3) the reactions to Reactive Oxygen Species; and (4) The resistance of mosquito larvae to three synthetic pesticides (temephos, imidacloprid, and permethrin). UV-B and Nitro-PAH treatment caused cellular damage and increased major detoxification enzymes such as α & β-esterase, cytoP450, CAT, GST, and POX. The levels of oxidative stress, ROS and protein carbonyl content, nitrite, ascorbic acid and thiobarbituric acid were decreased significantly. Toxicology bioassays revealed that UV-B + Nitro-PAH exposure significantly increased larval susceptibility. The current study concludes that prior exposure to Nitro-PAHs and UV-B may make mosquito larvae more vulnerable to chemical insecticides.

Back Cover: Gene Expression Profiling of 1α,25(OH)2D3 Treatment in 2D/3D Human Hepatocyte Models Reveals CYP3A4 Induction but Minor Changes in Other Xenobiotic‐Metabolizing Genes

Mol. Nutr. Food Res. 2022, 66, 202200070 DOI: 10.1002/mnfr.202200070 Few enzymes providing drug clearance are found under the control of activated vitamin D receptor (VDR) in the liver. In article number 2200070, Petr Pavek and co-workers investigate whether active vitamin D3 regulates major detoxification enzymes and transporters in 2D and 3D cultures of human hepatocytes. They show that VDR or vitamin D supplementation is unlikely to significantly influence drug elimination.

Proteomic analysis suggests that monoterpenes in lemongrass disrupt Ca2+ homeostasis in Haemaphysalis longicornis leading to mitochondrial depolarization and cytotoxicity.

Complex mixtures of bioactive ingredients in plant essential oils present complex chemistries which involve different modes of action. An increasing body of scientific reports has recently focused on the acaricidal activities of plant essential oils attributed to their monoterpene components, but information about their underlying molecular mechanism of action is scarce. Here, after the chemical analysis of lemongrass oil, a proteomic analysis of the ovary, salivary gland, and midgut of Haemaphysalis longicornis exposed to Cymbopogon citratus (lemongrass) essential oil was performed via data-independent acquisition mass spectrometry (DIA-MS) technology to further elucidate the molecular mechanisms involved. Pathway analysis reveals the activation of metabolic pathways mediated by oxidoreductases and transferases. Furthermore, the upregulation of various calcium-associated proteins and the upregulation of cytochrome c1, cytochrome c oxidase polypeptide IV, and programmed cell death protein 6-like isoform X1 suggest a cytotoxic mode of action via the formation of reactive oxygen species (ROS), mitochondrial Ca2+ overload, mitochondrial uncoupling, and depolarization, and ATP depletion leading to either apoptotic or necrotic death. Morphological alterations observed after the RNAi of a major detoxification enzyme (glutathione S-transferase) merit further investigation. Hence, the cytotoxic mode of action exhibited by C. citratus oil could be vital for the development of eco-friendly acaricide.

Biochemical Mechanism of Insecticide Resistance in Malaria Vector, Anopheles gambiae s.l in Nigeria.

Background:Malaria is a parasitic vector-borne disease endemic in the tropical and subtropical countries of the world. The aim of this study was to investigate the current activities of the detoxification enzymes in resistant and susceptible Anopheles gambiae s.l. in northern Nigeria.Methods:Anopheles larvae were collected from northeast and northwestern Nigeria between Aug and Nov 2018. Biochemical analyses was carried out on the mosquitoes exposed to various insecticides (deltamethrin, DDT, bendiocarb, malathion) to measure and compare the enzymatic activities of the major detoxification enzymes (P450, GSTs, Esterase).Results:High levels of resistance was observed; DDT 37%–53% (95%, CI: 29–61), bendiocarb 44%–55% (CI: 39–60) and deltamethrin 74%–82% (CI: 70–86). However, these mosquitoes were found to be susceptible to malathion 99%–100% (CI: 98–100). The P450 and GSTs enzymes were found to be elevated in the resistant mosquitoes exposed to deltamethrin (1.0240±0.1902); (1.3088±1.2478), DDT (1.7703±1.4528); (1.7462±0.9418) and bendiocarb (1.1814±0.0918); (1.4479±1.0083) compared to the Kisumu strain (0.764±0.4226); (0.6508±0.6542), (0.3875±0.3482); (0.4072±0.4916) and (0.6672±0.3949); (0.7126±0.7259) at P<0.05. Similarly, the resistant mosquitoes expressed increased activity to esterase (0.7606±1.1477), (0.3269±1.1957) and (2.8203±0.6488) compared to their susceptible counterpart (0.6841±0.7597), (0.7032±0.5380) and (0.6398±0.4159) at P<0.05. The enzyme ratio was found to be: P450 (1.341, 4.568 and 1.77); GSTs (2.011, 4.288 and 2.031); Esterases (1.111, 0.469 and 4.408). One way Anova and single sample t-test were also conducted to determine the effect of the enzymes on the resistant and susceptible strains.Conclusion:High level of insecticide resistance was observed with significant elevation of detoxification enzymes activities in the resistant mosquitoes.

Methylglyoxal-Induced Dysfunction in Brain Endothelial Cells via the Suppression of Akt/HIF-1α Pathway and Activation of Mitophagy Associated with Increased Reactive Oxygen Species.

Methylglyoxal (MG) is a dicarbonyl compound, the level of which is increased in the blood of diabetes patients. MG is reported to be involved in the development of cerebrovascular complications in diabetes, but the exact mechanisms need to be elucidated. Here, we investigated the possible roles of oxidative stress and mitophagy in MG-induced functional damage in brain endothelial cells (ECs). Treatment of MG significantly altered metabolic stress as observed by the oxygen-consumption rate and barrier-integrity as found in impaired trans-endothelial electrical resistance in brain ECs. The accumulation of MG adducts and the disturbance of the glyoxalase system, which are major detoxification enzymes of MG, occurred concurrently. Reactive oxygen species (ROS)-triggered oxidative damage was observed with increased mitochondrial ROS production and the suppressed Akt/hypoxia-inducible factor 1 alpha (HIF-1α) pathway. Along with the disturbance of mitochondrial bioenergetic function, parkin-1-mediated mitophagy was increased by MG. Treatment of N-acetyl cysteine significantly reversed mitochondrial damage and mitophagy. Notably, MG induced dysregulation of tight junction proteins including occludin, claudin-5, and zonula occluden-1 in brain ECs. Here, we propose that diabetic metabolite MG-associated oxidative stress may contribute to mitochondrial damage and autophagy in brain ECs, resulting in the dysregulation of tight junction proteins and the impairment of permeability.

Effect of age on insecticide susceptibility and enzymatic activities of three detoxification enzymes and one invertase in honey bee workers (Apis mellifera)

Honey bee is an economically important insect for honey production and pollination. Frequent exposure to toxic pesticides is one of the major risk factors causing the pollinator population decline. However, age effects of honey bees on pesticide susceptibility have been largely ignored and many researchers use bees of unknown age for assessing the risk of pesticides. Honey bee workers are known to go through physiological and behavioral changes in order to differentiate different phenotypes to perform specific duties over their natural lifetime of 6 weeks or longer. In this study, we provide multi-parameter evidences of unignorable age effects of honey bee workers and suggest using a standard bee age to produce reliable and comparable data when assessing variable and realistic situations of in-hive and field exposures to pesticides. Using honey bee workers aged 4- to 42-days old, we examined susceptibility of the bees to five different insecticides from five different classes and measured enzymatic activities of three major detoxification enzymes and an invertase involved in honey production. Results showed gradual increase of natural mortality and decrease of soluble protein content in bees over the age span from 4 days to 42 days. Significant increases of mortality after separate treatments of five different insecticides confirmed drastic age effects of bees over the assessed age span. As they aged, honey bees also showed a gradual increase of cytochrome P450 oxidase activity while still maintaining constant levels of two other detoxification enzymes (esterase and glutathione S-transferase) and an invertase responsible for honey production.

&lt;p&gt;&lt;strong&gt;&lt;span&gt;Transcriptome analys&lt;/span&gt;&lt;/strong&gt;&lt;strong&gt;&lt;span&gt;i&lt;/span&gt;&lt;/strong&gt;&lt;strong&gt;&lt;span&gt;s of &lt;/span&gt;&lt;/strong&gt;&lt;strong&gt;&lt;em&gt;&lt;span&gt;Tetranychus cinnabarinus&lt;/span&gt;&lt;/em&gt;&lt;/strong&gt;&lt;strong&gt;&lt;span&gt; responses to an insecticide exposure&lt;/span&gt;&lt;/strong&gt;&lt;/p&gt;

Diflubenzuron, a benzoylphenylurea insecticide that interferes with chitin biosynthesis, causes arthropods to moult abnormally and die. However, its mechanism of action in Tetranychus cinnabarinus is still unclear. In order to explore the effects of different sublethal concentrations of diflubenzuron on T. cinnabarinus, we conducted a high-throughput RNA-seq technology to identify the variations in transcriptomic profile of T. cinnabarinus larvae. The results revealed that 470 and 49 differentially expressed genes were identified in LC50-and LC70-treated groups, comparing with the control. We also identified and analyzed the detoxification enzymes involved in the transcritome of T. cinnabarinus, including 34 cytochrome P450 genes, 17 glutathione-s-transferase genes (GSTs), 12 acetylcholinesterase genes (AChEs) and 9 ABC transporter genes. In addition, differentially expressed genes analysis showed that the gene expression levels of detoxification enzymes were generally enhanced. At the same time, seven and eleven genes were involved in chitin synthesis and degradation ways, respectively. The expression level of most genes involved in chitin synthesis and degradation pathway were generally up-regulated after exposure to sublethal concentrations of diflubenzuron. Moreover, for transcriptome validation, the mRNA expression results of ten specially expressed genes by quantitative real-time PCR demonstrated that these gene expression trends were consistent with that of the transcriptome data. Together, all these results suggested that sublethal concentrations of diflubenzuron exposure affected gene expression of major detoxification enzymes and chitin metabolism genes in T. cinnabarinus larvae. These findings may be helpful to further understand the possible molecular mechanism of benzoylphenylurea insecticides in T. cinnabarinus, as well as in other spider mites.

Carboxylesterase genes in nitenpyram-resistant brown planthoppers, Nilaparvata lugens.

Carboxylesterases (CarEs) represent one of the major detoxification enzyme families involved in insecticide resistance. However, the function of specific CarE genes in insecticide resistance is still unclear in the insect Nilaparvata lugens (Stål), a notorious rice crop pest in Asia. In this study, a total of 29 putative CarE genes in N. lugens were identified, and they were divided into seven clades; further, the β-esterase clade was significantly expanded. Tissue-specific expression analysis found that 17 CarE genes were abundantly distributed in the midgut and fat body, while 12 CarE genes were highly expressed in the head. The expression of most CarE genes was significantly induced in response to the challenge of nitenpyram, triflumezopyrim, chlorpyrifos, isoprocarb and etofenprox. Among these, the expression levels of NlCarE2, NlCarE4, NlCarE9, NlCarE17 and NlCarE24 were increased by each insecticide. Real-time quantitative polymerase chain reaction and RNA interference assays revealed the NlCarE1 gene to be a candidate gene mainly involved in nitenpyram resistance, while simultaneously silencing NlCarE1 and NlCarE19 produced a stronger effect than silencing either one individually, suggesting a cooperative relationship in resistance formation. These findings lay the foundation for further clarification of insecticide resistance mediated by CarE in N. lugens.

Convergent evolution of conserved mitochondrial pathways underlies repeated adaptation to extreme environments

Extreme environments test the limits of life; yet, some organisms thrive in harsh conditions. Extremophile lineages inspire questions about how organisms can tolerate physiochemical stressors and whether the repeated colonization of extreme environments is facilitated by predictable and repeatable evolutionary innovations. We identified the mechanistic basis underlying convergent evolution of tolerance to hydrogen sulfide (H2S)-a toxicant that impairs mitochondrial function-across evolutionarily independent lineages of a fish (Poecilia mexicana, Poeciliidae) from H2S-rich springs. Using comparative biochemical and physiological analyses, we found that mitochondrial function is maintained in the presence of H2S in sulfide spring P. mexicana but not ancestral lineages from nonsulfidic habitats due to convergent adaptations in the primary toxicity target and a major detoxification enzyme. Genome-wide local ancestry analyses indicated that convergent evolution of increased H2S tolerance in different populations is likely caused by a combination of selection on standing genetic variation and de novo mutations. On a macroevolutionary scale, H2S tolerance in 10 independent lineages of sulfide spring fishes across multiple genera of Poeciliidae is correlated with the convergent modification and expression changes in genes associated with H2S toxicity and detoxification. Our results demonstrate that the modification of highly conserved physiological pathways associated with essential mitochondrial processes mediates tolerance to physiochemical stress. In addition, the same pathways, genes, and-in some instances-codons are implicated in H2S adaptation in lineages that span 40 million years of evolution.

The Interaction of Schistosoma Japonicum Glutathione Transferase with Cibacron Blue 3GA and its Fragments.

The 26kDa glutathione transferase (GST, EC 2.5.1.18) from Schistosoma japonicum (SjGST) is recognized as the major detoxification enzyme of S. japonicum, a pathogenic helminth causing schistosomiasis. In the present study, the interaction of the chlorotriazine dye Cibacron blue 3GA (CB3GA) and its structural analogues with SjGST was investigated. The work aimed to shed light on the non-substrate ligand-binding properties of the enzyme. Kinetic inhibition analysis, affinity labelling experiments and molecular modelling studies were employed. The results showed that CB3GA is a potent inhibitor (IC50 0.057 ± 0.003 μM) towards SjGST. The enzyme was specifically and irreversibly inactivated by the dichlorotriazine-analogue of CB3GA (IC50 0.190 ± 0.024 μM), following a biphasic pseudo-first-order saturation kinetics with approximately 1 mol of inhibitor per mol of the dimeric enzyme being incorporated. All other monochlorotriazine analogues behave as reversible inhibitors with lower inhibition potency (IC50 5.2-82.3 μM). Kinetic inhibition studies, together with molecular modelling and molecular dynamics simulations, established that the CB3GA binding site overlaps both the G- and H-sites. Both hydrophobic/ polar interactions, as well as steric effects, have decisive roles in determining the inhibitory strength of CB3GA and its analogues. The results of the present study might be useful in future drug design and development efforts towards SjGST.

Virtual screening and docking of lead like molecules against Glutathione-S-Transferase protein from Brugia malayi.

Glutathione-S-transferase(s) (GST) is an important chemotherapeutic target in lymphatic filarasis caused by Brugia malayi and Wuchereria bancrofti. It has been playing an important role as major detoxification enzyme and help in intracellular transportation of hydrophobic substrates. Therefore, it is of interest to screen GST from Brugia malayi with millions of known ligands at the ZINC database using AUTODOCK for the identification of potential inhibitors with improved binding characteristics. We report two potent inhibitors ZINC00179016 and ZINC08385519 which are the molecules of pyrrolidinedione and benzimidazole families respectively as potential inhibitors of GST from Brugia malayi with suitable binding properties.

Stable integration of the Mrx1-roGFP2 biosensor to monitor dynamic changes of the mycothiol redox potential in Corynebacterium glutamicum

Mycothiol (MSH) functions as major low molecular weight (LMW) thiol in the industrially important Corynebacterium glutamicum. In this study, we genomically integrated an Mrx1-roGFP2 biosensor in C. glutamicum to measure dynamic changes of the MSH redox potential (EMSH) during the growth and under oxidative stress. C. glutamicum maintains a highly reducing intrabacterial EMSH throughout the growth curve with basal EMSH levels of ~− 296 mV. Consistent with its H2O2 resistant phenotype, C. glutamicum responds only weakly to 40 mM H2O2, but is rapidly oxidized by low doses of NaOCl. We further monitored basal EMSH changes and the H2O2 response in various mutants which are compromised in redox-signaling of ROS (OxyR, SigH) and in the antioxidant defense (MSH, Mtr, KatA, Mpx, Tpx). While the probe was constitutively oxidized in the mshC and mtr mutants, a smaller oxidative shift in basal EMSH was observed in the sigH mutant. The catalase KatA was confirmed as major H2O2 detoxification enzyme required for fast biosensor re-equilibration upon return to non-stress conditions. In contrast, the peroxiredoxins Mpx and Tpx had only little impact on EMSH and H2O2 detoxification. Further live imaging experiments using confocal laser scanning microscopy revealed the stable biosensor expression and fluorescence at the single cell level. In conclusion, the stably expressed Mrx1-roGFP2 biosensor was successfully applied to monitor dynamic EMSH changes in C. glutamicum during the growth, under oxidative stress and in different mutants revealing the impact of Mtr and SigH for the basal level EMSH and the role of OxyR and KatA for efficient H2O2 detoxification under oxidative stress.

Virtual screening of natural anti-filarial compounds against glutathione-S-transferase of Brugia malayi and Wuchereria bancrofti

Glutathione-S-transferase also referred as GST is one of the major detoxification enzymes in parasitic helminths. The crucial role played by GST in various chronic infections has been well reported. The dependence of nematodes on detoxification enzymes to maintain their survival within the host established the crucial role of GST in filariasis and other related diseases. Hence, this well-established role of GST in filariasis along with its greater nonhomology with its human counterpart makes it an important therapeutic drug target. Here in this study, we have tried to explore the inhibitory potential of some of the well-reported natural ant-filarial compounds against the GST from Wuchereria bancrofti (W.bancrofti) and Brugia malayi (B.malayi). In silico virtual screening, approach was used to screen the selected natural compounds against GST from W.bancrofti and B.malayi. On the basis of our results, here we are reporting some of the natural compounds which were found to be very effective against GSTs. Along with we have also revealed the characteristic of the active site of BmGST and WbGST and the role of important active site residues involve in the binding of natural compounds within the active site of GSTs. This information will oped doors for using natural compounds as anti-filarial therapy and will also be helpful for future drug discovery.

Gossypol-induced fitness gain and increased resistance to deltamethrin in beet armyworm, Spodoptera exigua (Hübner).

In plant-insect interactions, phytotoxins such as gossypol, exert a defensive role on behalf of the plant by interfering with the essential metabolic, biochemical and physiological pathways of herbivorous insects. The beet armyworm, Spodoptera exigua (Hübner), is a key pest for many important crops including a range of vegetables, ornamentals, and cotton. In this study, we investigated how resistance to deltamethrin relates to enzyme activity in gossypol-pretreated larvae of S. exigua. Following selection with deltamethrin insecticides on gossypol-pretreated larvae for 10 generations, the Gos-SEL population developed a 113.29-fold resistance. Under the same conditions, the Delta-SEL selected population showed a 69.76-fold increase in resistance along with corresponding levels of xenobiotic defense enzyme activity. Similarly, the fecundity of the Delta-SEL population together with male and female longevity were found to be significantly lower when compared with the Gos-SEL population and the laboratory susceptible-strain group (SS-Strain). In addition, the activities of cytochrome P450s in S. exigua were significantly enhanced when the insects were fed on a deltamethrin and gossypol-pretreated diet compared with being fed on deltamethrin alone. The reproductive capacity of S. exigua is significantly reduced in Delta-SEL and Gos-SEL populations compared with the control group (SS-Strain). Elevation of the major detoxification enzyme cytochrome P450 monooxygenase and esterase might have an important role in inducing tolerance to deltamethrin in gossypol-fed S. exigua populations. This study enhances our understanding of detoxification enzyme pathways for S. exigua gene expression and their role in responses to insecticides and plant secondary metabolites. © 2018 Society of Chemical Industry.

Biochemical and molecular characterizations of cypermethrin resistance in laboratory-selected cypermethrin-resistant strains of Tetranychus urticae Koch. (Acari: Tetranychidae)

ABSTRACTPyrethroids are commonly used pesticides for controlling agricultural pests. A pyrethroid resistant strain of T. urticae was obtained after laboratory selection with cypermethrin for determining mechanisms of cypermethrin resistance. TaqMan diagnostic assay as well as sequencing of voltage gated sodium channel (VGSC) gene were performed in order to detect previously known mutations in German susceptible strain (GSS) confirming that it does not carry any of the known VGSC mutations. Therefore, GSS strain was selected with cypermethrin and the resistant C6 strain was generated that is free of VGSC mutations. Toxicity assay data revealed relatively high resistance levels of the C6 strain (54.24-folds) in comparison to GSS. All inhibitors (piperonyl butoxide (PBO), esterases, P450; S-benzyl-O, O-diisopropylphosphorothioate (IBP), esterases; and diethyl maleate (DEM), glutathione S-transferase (GST)) were used in combined toxicity assays against the major detoxification enzymes, synergized resistance by 1.41-, 1.61-, and 2.27-fold, respectively. In the C6 strain, activities of P450, GST, and esterase showed elevated activity in comparison to GSS. By application of TaqMan diagnostic assay, we confirmed that both GSS and C6 T. urticae strains are free of VGSC mutations, indicating that detoxification-based cypermethrin resistance mechanisms are involved in resistance and have significant effect on the phenotype.

Spermidine Enhances Activities of Detoxification Enzymes in Onion (&amp;lt;i&amp;gt;Allium cepa&amp;lt;/i&amp;gt; L.) Seedlings Under Short Term Salinity

In plant, glyoxalases [glyoxalase I (Gly-I, EC: 4.4.1.5) and glyoxalase II (Gly-II, EC: 3.1.2.6)] and glutathione S-transferase (GST, EC: 2.5.1.18) are major detoxification enzymes. On the other hand, spermidine (Spd) is important polyamine (PA) with significant role which interacts with stress protection mechanisms functioning in common against different types of stress. In this study, exogenous Spd was applied on onion seedlings to investigate its protective role through regulation of glyoxalase and GST activities. Continuous increase was observed in the content of methylglyoxal (MG) in onion leaves under salinity, and at 7 day of stress, MG contents increased by 260% over control. Application of Spd reduced the MG contents in saline treated seedlings through increasing glyoxalase mediated detoxification by 21 and 48% at 1 and 3 day of stress, respectively. Salinity increased Gly-I and Gly-II activities which was further increased by Spd upto 3 day of stress. On the other hand, salinity increased GST activity by 14, 55, 93 and 109% over control at 1, 3, 5 and 7 day, respectively. Application of Spd increased the activity in stressed seedlings at 3 day of stress while 21% higher activity was found. However, after 3 days, both glyoxalases and GST activities in Spd treated seedlings decreased and became almost similar to those in drought stressed seedlings without Spd. Considering the results, application of Spd in onion seedlings improved tolerance for short period of salinity.

Incidence and characterization of resistance to pyrethroid and organophosphorus insecticides in Thrips tabaci (Thysanoptera: Thripidae) in onion fields in Isfahan, Iran

Onion thrips, Thrips tabaci Lindeman, is the main pest of onion-growing fields in Isfahan and is mainly controlled by frequently spraying several insecticides. To investigate the resistance status and mechanisms, the susceptibility of ten field populations collected from Isfahan onion-growing regions were tested to several currently used pesticides. Resistance to the tested insecticides was observed in most populations when compared with the susceptible reference population. Enhanced detoxification, implicated by the use of inhibitors of major metabolic detoxification enzymes, was observed in the populations resistant to profenofos and chlorpyrifos. In the deltamethrin resistant populations, the amino acid substitution T929I was detected in the voltage gated sodium channel, which is known to confer pyrethroid resistance. These data are a first step towards more efficient resistance management tactics through early detection of resistant onion thrips in Iran.