Functional Characterization of Two Carboxylesterase Genes Involved in Pyrethroid Detoxification in <i>Helicoverpa armigera</i>

Li, Yongqiang; Bai, Leichao; Zhao, Chen; Xu, Jian; Sun, Zhongjuan; Dong, Yanling; Li, Dexian; Liu, Xili; Ma, Zhiqing

doi:10.1021/acs.jafc.9b06690

Cited by 28 publications

(28 citation statements)

References 49 publications

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“…Carboxylesterases (CarEs, EC 3.1.1.1) are considered one of the most important phase I detoxifying enzymes involved in the detoxification of many types of insecticides including organophosphates, pyrethroids and carbamates, [4][5][6][7] and CarEs play crucial roles in insecticide resistance in many insect pests. [8][9][10][11] Numerous studies have shown that mutation and overexpression of related genes are two major mechanisms for CarE-mediated insecticide resistance.…”

Section: Introductionmentioning

confidence: 99%

Functional analysis of a carboxylesterase gene involved in beta‐cypermethrin and phoxim resistance in Plutella xylostella (L.)

Zhu

Shan

et al. 2021

Pest Management Science

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BACKGROUND Carboxylesterases (CarEs) are associated with detoxification of xenobiotics, including insecticides, in organism bodies. Overexpression of CarE genes is considered to have an important role in insecticide resistance in insects, however its involvement in multi‐insecticide resistance has rarely been reported. This study aimed to assess the function of a CarE gene (PxαE8) in resistance to five insecticides in Plutella xylostella. RESULTS Relative expression of PxαE8 in three multi‐insecticide‐resistant Plutella xylostella populations, GD‐2017, GD‐2019 and HN, was14.8‐, 19.5‐ and 28.0‐fold higher than that in the susceptible population. Exposure to lethal concentrations associated with 25% mortality (LC25) of beta‐cypermethrin, chlorantraniliprole, metaflumizone, phoxim and tebufenozide could induce the specific activity of CarEs and increase the relative expression of PxαE8. By contrast, knockdown of PxαE8 expression dramatically reduced the activity of CarEs and increased the resistance of P. xylostella (GD‐2019) larvae to beta‐cypermethrin and phoxim by 47.4% and 45.5%, respectively. Further, a transgenic line of Drosophila melanogaster overexpressing PxαE8 was constructed and the bioassay results showed that the tolerance of transgenic Drosophila to beta‐cypermethrin and phoxim was 3.93‐ and 3.98‐fold higher than that of the untransgenic line. CONCLUSION These results provide evidence that overexpression of PxαE8 is involved in resistance, at least to beta‐cypermethrin and phoxim, in multi‐insecticide‐resistant P. xylostella populations, which could help in further understanding the molecular mechanisms of multi‐insecticide resistance in this pest. © 2020 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 99%

Functional analysis of a carboxylesterase gene involved in beta‐cypermethrin and phoxim resistance in Plutella xylostella (L.)

Zhu

Shan

et al. 2021

Pest Management Science

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“…In 2013, a study which involved the heterologous expression of CarEs from H. armigera using a baculovirus system, firstly reported that individual CarEs had relatively low hydrolase activity toward either OPs or SPs 44 . Similarly in our previous studies, heterologous expression in the Escherichia coli demonstrated that a few CarEs, including CarE001D, CarE001G, CarE001A and CarE001H from H. armigera strain (WH), had metabolic activities toward SPs 45–47 . Particularly CarE001G exhibited highly catalytic efficiency toward α ‐naphthyl acetate ( α ‐NA), strong binding affinity to OPs, and relatively low hydrolase activity toward three types of SPs.…”

Section: Introductionmentioning

confidence: 89%

“…The DNA of variants of CarE001G were amplified with genespecific primers containing restriction endonuclease recognition sites (Supporting Information, Table S1) by using the PrimeSTAR HS DNA Polymerase (TaKaRa, Shiga, Japan) following the method of Li et al 47 with slight modification. Briefly, the amplification program was 98°C for 30 s, followed by 30 cycles of 98°C for 12 s, 68°C for 25 s, 72°C for 2.5 min, and with a final extension of 72°C for 5 min.…”

Section: Construction and Bacterial Expression Of Mutant Enzymesmentioning

confidence: 99%

“…To characterize the function of these mutant CarEs, the sequencing-verified plasmids were used to yield mutant CarEs according to the protocol of our previous report. 46,47 The protein expression was induced by isopropylthio-galactoside (IPTG) at a final concentration of 0.2 mmol L -1 at 18°C for 48 h with shaking at 200 rpm.…”

Section: Construction and Bacterial Expression Of Mutant Enzymesmentioning

confidence: 99%

“…44 Similarly in our previous studies, heterologous expression in the Escherichia coli demonstrated that a few CarEs, including CarE001D, CarE001G, CarE001A and CarE001H from H. armigera strain (WH), had metabolic activities toward SPs. [45][46][47] Particularly CarE001G exhibited highly catalytic efficiency toward ⊍-naphthyl acetate (⊍-NA), strong binding affinity to OPs, and relatively low hydrolase activity toward three types of SPs. Furthermore, CarE001G has a longer open reading frame (ORF) encoding 747 amino acid (aa) residues than most of other CarEs (550-560 aa), and contains a glycine-rich region (GRR) (approximately 60% glycine residues) of 146 aa located adjacent to the C-terminal.…”

Section: Introductionmentioning

confidence: 99%

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Enhanced hydrolysis of β‐cypermethrin caused by deletions in the glycin‐rich region of carboxylesterase 001G from Helicoverpa armigera

Bai

Zhao

et al. 2021

Pest Management Science

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BACKGROUND Carboxylesterase (CarE) is a major class of enzyme involved in the detoxification of toxic xenobiotics in various insect species. Previous work has shown that the carboxylesterase gene CarE001G found in Helicoverpa armigera is more active and can metabolize synthesized pyrethroids, such as β‐cypermethrin, one of the commonly used commercial insecticides for lepidopteran pest control. In addition, CarE001G is very special as it has a very specific glycine‐rich region located adjacent to its C‐terminal. But whether mutations in this unique sequence can change the biochemistry and function of CarE001G are unknown. RESULTS In this study, four variants of CarE001G with different deletions in the glycine‐rich region were obtained and functionally expressed in Escherichia coli. The recombinant proteins were purified and confirmed by Western blot and mass spectrometry analyses. These mutant enzymes showed high catalytic efficiency toward the model substrate α‐naphthyl acetate. Inhibition study showed that β‐cypermethrin had relatively strong inhibition on CarE activities. In vitro metabolism assay showed that the mutant enzymes significantly enhanced their metabolic activities toward β‐cypermethrin with specific activities between 4.0 and 5.6 nmol L‐1 min−1 mg−1 protein. Molecular docking analyses consistently demonstrated that deletion mutations in the glycine‐rich region may facilitate the anchoring of the β‐cypermethrin molecule in the active binding pocket of the mutant enzymes. CONCLUSION The data show that deletion mutations can cause qualitative change in the capacity of CarEs in the detoxification of β‐cypermethrin. This indicates that deletion mutations in the glycine‐rich region may have the potential to cause synthesized pyrethroid (SP) resistance in H. armigera in the future. © 2020 Society of Chemical Industry

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The effects of quercetin combined with nucleopolyhedrovirus on the growth and immune response in the silkworm (Bombyx mori)

Guo-qing

Kang

Liu

et al. 2021

Arch Insect Biochem Physiol

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Flavonoids are secondary metabolites that help plants resist insect attack. It can resist insect attack by inhibiting insect immune defense, and pathogens can also inhibit insect immune defense. It is speculated that the combination of flavonoids and pathogens may inhibit the immune defense and have stronger toxicity to silkworm. In this study, the combined treatment of quercetin with Bombyx mori nuclear polyhedrosis virus (BmNPV) had significant negative effects on the growth and survival of silkworm compared with BmNPV group. The detoxifying enzyme activity of BmNPV group was significantly increased at 96 h, while the activity of the combined treatment group was significantly decreased with the increase of quercetin exposure time (72 or 96 h). The activity of antioxidant enzymes also showed a similar trend, that was, the activity of antioxidant enzymes in the combined treatment group also decreased significantly with the increase of quercetin exposure time, which led to the increase of reactive oxygen species content. The silkworm cells would produce lipid peroxidation, malondialdehyde content was significantly increased, so that the expression of immune‐related genes (the antimicrobial peptide, Toll pathway, IMD pathway, JAK‐STAT pathway, and melanin genes) were decreased, leading to the damage of the immune system of silkworm. These results indicated that quercetin combined with BmNPV could inhibit the activities of protective enzymes and lead to oxidative damage to silkworm. It can also affect the immune response of the silkworm, and thus resulting in abnormal growth. This study provides the novel conclusion that quercetin accumulation will increase the susceptibility of silkworm to pathogens.

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Functional Characterization of Two Carboxylesterase Genes Involved in Pyrethroid Detoxification in Helicoverpa armigera

Cited by 28 publications

References 49 publications

Functional analysis of a carboxylesterase gene involved in beta‐cypermethrin and phoxim resistance in Plutella xylostella (L.)

Functional analysis of a carboxylesterase gene involved in beta‐cypermethrin and phoxim resistance in Plutella xylostella (L.)

Enhanced hydrolysis of β‐cypermethrin caused by deletions in the glycin‐rich region of carboxylesterase 001G from Helicoverpa armigera

The effects of quercetin combined with nucleopolyhedrovirus on the growth and immune response in the silkworm (Bombyx mori)

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