CRISPR/Cas9 mediated G4946E substitution in the ryanodine receptor of Spodoptera exigua confers high levels of resistance to diamide insecticides

Zuo, Yayun; Wang, Hui; Xu, Yanjun; Huang, Jianlei; Wu, Shuwen; Wu, Yingliang; Yang, Yihua

doi:10.1016/j.ibmb.2017.09.005

Cited by 99 publications

(80 citation statements)

References 48 publications

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“…In diamondback moth, tomato leafminer and rice stem borer, it has been shown that diamide resistance is conferred by RyR target‐site mutations leading to amino acid substitutions at two major sites, G4946 and I4790 . It has been shown using radioligand binding studies, cell‐based studies and CRISPR/Cas9 genome editing that these mutations are functional in conferring diamide resistance . However, the molecular mechanisms of high levels of diamide resistance (RR > 100 000‐fold) in beet armyworm, S. exigua remain unknown.…”

Section: Discussionmentioning

confidence: 99%

“…The recently described G4946E substitution – not detected in Chlorant‐R RyR – has been functionally linked to high levels of diamide resistance in at least two lepidopteran pests, P. xylostella and T. absoluta . Recently, the G4946E mutation has been introduced via CRISPR/Cas9 technology in S. exigua and resulted in high levels of diamide resistance in transgenic beet armyworm …”

Section: Discussionmentioning

confidence: 99%

“…The most important mechanism of resistance in diamondback moth has been functionally linked to target‐site mutations in the RyR transmembrane domain, such as the amino acid substitutions G4946E and I4790M . The functional relevance of these RyR mutations was not confirmed not only by radioligand binding studies using flight muscle microsomal membrane preparations of diamondback moth and tomato leafminer adults, but also by recombinant expression of mutant RyR variants in insect cell lines, and by CRISPR/Cas9 genome‐edited transgenic Drosophila melanogaster and S. exigua carrying RyR M4790I and G4946E mutations, respectively . Both amino acid residues have also been shown to be mutated in diamide‐resistant tomato leafminer and rice stem borers …”

Section: Introductionmentioning

confidence: 99%

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Detection of a ryanodine receptor target‐site mutation in diamide insecticide resistant fall armyworm, Spodoptera frugiperda

Boaventura

Bolzan

Padovez

et al. 2019

Pest Management Science

117

126

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BACKGROUND Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), a major lepidopteran pest in Latin and North America, has very recently invaded the continents of Africa and Asia. FAW has evolved resistance to different insecticides and transgenic corn expressing Bacillus thuringiensis (Bt) toxins. Here, we investigated the extent and mechanisms of resistance to diamide insecticides in a Brazilian field‐collected FAW strain selected using chlorantraniliprole. RESULTS Continuous laboratory selection of a field‐collected FAW strain with chlorantraniliprole resulted in resistance ratios of 225‐fold and > 5400‐fold against chlorantraniliprole and flubendiamide, respectively, when compared with a susceptible strain. Pre‐exposure to different synergists known to inhibit detoxification enzymes did not result in significantly increased larval toxicity, suggesting a minor role for metabolic resistance. Sequencing of the FAW ryanodine receptor (RyR) C‐terminal domains II to VI revealed a single nucleotide polymorphism, resulting in a I4734M mutation recently said to confer target‐site resistance to diamides in lepidopteran pests. Genotyping by pyrosequencing of field‐collected FAW larvae sampled in the 2018 crop season suggests a low resistance allele frequency. Furthermore, we developed a fluorescent polymerase chain reaction (PCR)‐based allelic discrimination assay for rapid genotyping of field‐collected FAW samples, because diamides are increasingly used in Bt−/non‐Bt corn. CONCLUSIONS Recently, the identified RyR mutation has been shown to confer field resistance in other lepidopteran pests such as diamondback moth, tomato leafminer and striped rice stem borer. The developed PCR‐based allelic discrimination assay will help to monitor the frequency and future spread of diamide resistance allele in FAW field populations and help to implement appropriate resistance management measures. © 2019 Society of Chemical Industry

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Detection of a ryanodine receptor target‐site mutation in diamide insecticide resistant fall armyworm, Spodoptera frugiperda

Boaventura

Bolzan

Padovez

et al. 2019

Pest Management Science

117

126

View full text Add to dashboard Cite

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“…The CRISPR/Cas9 system has been successfully applied in a number of insect species, and has recently been used to investigate insecticide resistance mechanisms . To date, by using CRISPR/Cas9 coupled with non‐homologous end joining (NHEJ) and/or homology directed repair (HDR) approaches, the following mutation(s) edited in the nAChRα6 subunit (G275E and P146S of D. melanogaster ), chitin synthase 1 ( CHS1 , I1015M/F/L of D. melanogaster ), ryanodine receptors ( RyR , G4946E/V & M4790I of D. melanogaster and Spodoptera exigua ), cytochrome P450 gene ( CYP9M10 , KO in Culex quinquefasciatus ), cadherin ( CAD , KO in Helicoverpa armigera ), P‐glycoprotein ( P‐gp , KO in S. exigua ), ABCA2 (KO & KI in H. armigera ), and tetraspanin ( TSPAN1 , KO & KI in H. armigera ) were confirmed in vivo to be involved in insect susceptibility to chemical insecticides or Bacillus thuringiensis Cry toxins …”

Section: Introductionmentioning

confidence: 99%

Disruption of nicotinic acetylcholine receptor α6 mediated by CRISPR/Cas9 confers resistance to spinosyns in Plutella xylostella

Wang

et al. 2019

Pest Management Science

Self Cite

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BACKGROUND The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system provides some advantages over other reverse genetic techniques to investigate the causal relationship between insecticide resistance phenotype and candidate gene. Several studies published to date point to the nicotinic acetylcholine receptor (nAChR) α6 subunit strongly associated with spinosyns resistance in insects, including Plutella xylostella. However, reverse genetic verification of the P. xylostella nAChRα6 has not yet been achieved via an in vivo approach. RESULTS Here, we successfully constructed a homozygous strain (Pxα6‐KO) with a 2‐nt deletion mutation of nAChRα6 by CRISPR/Cas9 coupled with non‐homologous end joining approach in P. xylostella. The manipulated mutation results in a frame shift in the open reading frame of transcripts, which produces a predicted protein truncated in the TM3–TM4 loop region. When compared to the background strain IPP‐S, the knockout strain Pxα6‐KO exhibited 229‐ and 1462‐fold resistance to spinosad and spinetoram, respectively, but no or limited (resistance ratios <3‐fold) effects on the toxicities of imidacloprid, abamectin, β‐cypermethrin, indoxacarb, metaflumizone and chlorantraniliprole. Furthermore, the mode of inheritance of the acquired spinetoram resistance was autosomal recessive and significantly linked with the 2‐nt deletion mutation of nAChRα6 in the Pxα6‐KO strain. CONCLUSION In vivo functional investigation demonstrates the causality of the Pxα6 truncating mutation with high levels of resistance to spinosyns in P. xylostella. Our results suggest the Pxα6‐KO strain underlies an autosomal, recessive mode of inheritance for spinetoram resistance, and reinforces the association of this gene to the mode of action of spinosyns. © 2019 Society of Chemical Industry

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“…2,3 Currently, because of the generation of resistance and cross resistance, the insecticidal effect of phthaldiamides (such as uobendiamide) and traditional organophosphorus pesticides was tapered. [4][5][6][7][8] Thus, the development of insecticidal agent with novel structure is increasingly required.…”

Section: Introductionmentioning

confidence: 99%

Design, synthesis, insecticidal activity and 3D-QSR study for novel trifluoromethyl pyridine derivatives containing an 1,3,4-oxadiazole moiety

Wang

Luo

et al. 2018

RSC Adv.

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A series of trifluoromethyl pyridine derivatives containing 1,3,4-oxadiazole moiety was designed, synthesized and bio-assayed for their insecticidal activity. The result of bio-assays indicated the synthesized compounds exhibited good insecticidal activity against Mythimna separata and Plutella xylostella, most of the title compounds show 100% insecticidal activity at 500 mg L À1 and >80% activity at 250 mg L À1 against the two pests. Compounds E18 and E27 showed LC 50 values of 38.5 and 30.8 mg L À1 against Mythimna separata, respectively, which were close to that of avermectin (29.6 mg L À1 ); compounds E5, E6, E9, E10, E15, E25, E26, and E27 showed 100% activity at 250 mg L À1, which were better than chlorpyrifos (87%). CoMFA and CoMSIA models with good predictability were proposed, which revealed the electron-withdrawing groups with an appropriate bulk at 2-and 4-positions of benzene ring could enhance insecticidal activity.

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CRISPR/Cas9 mediated G4946E substitution in the ryanodine receptor of Spodoptera exigua confers high levels of resistance to diamide insecticides

Cited by 99 publications

References 48 publications

Detection of a ryanodine receptor target‐site mutation in diamide insecticide resistant fall armyworm, Spodoptera frugiperda

Detection of a ryanodine receptor target‐site mutation in diamide insecticide resistant fall armyworm, Spodoptera frugiperda

Disruption of nicotinic acetylcholine receptor α6 mediated by CRISPR/Cas9 confers resistance to spinosyns in Plutella xylostella

Design, synthesis, insecticidal activity and 3D-QSR study for novel trifluoromethyl pyridine derivatives containing an 1,3,4-oxadiazole moiety

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