Molecular characterization of Cry1F resistance in fall armyworm, Spodoptera frugiperda from Brazil

Boaventura, Debora; Ulrich, Julia; Lueke, Bettina; Bolzan, Aline Adriana; Okuma, Daniela M.; Gutbrod, Oliver; Geibel, Sven; Zeng, Qin; Dourado, Patrick M.; Martinelli, Samuel; Flagel, Lex; Head, Graham P.; Nauen, Ralf

doi:10.1016/j.ibmb.2019.103280

Cited by 72 publications

(111 citation statements)

References 60 publications

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“…R 1 was not detected in Cry1Fa-resistant populations in Florida, the Dominican Republic, or Brazil; and a second resistance allele (R 2 ) of SfABCC2 was isolated from the field in Puerto Rico, but its frequency was not reported [38,39]. A recent study identified many mutations in SfABCC2 associated with field-selected resistance to Cry1Fa in Brazil [50]. As far as we know, mutations in ABCC3 were not evaluated in the cases of field-evolved resistance in T. ni and S. frugiperda cited above.…”

Section: Plos Pathogensmentioning

confidence: 99%

Functional redundancy of two ABC transporter proteins in mediating toxicity of Bacillus thuringiensis to cotton bollworm

Wang

Zhao

et al. 2020

PLoS Pathog

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Evolution of pest resistance reduces the efficacy of insecticidal proteins from the gram-positive bacterium Bacillus thuringiensis (Bt) used widely in sprays and transgenic crops. Better understanding of the genetic basis of resistance is needed to more effectively monitor, manage, and counter pest resistance to Bt toxins. Here we used CRISPR/Cas9 gene editing to clarify the genetics of Bt resistance and the associated effects on susceptibility to other microbial insecticides in one of the world's most damaging pests, the cotton bollworm (Helicoverpa armigera). We discovered that CRISPR-mediated knockouts of ATP-binding cassette (ABC) transporter genes HaABCC2 and HaABCC3 together caused >15,000-fold resistance to Bt toxin Cry1Ac, whereas knocking out either HaABCC2 or HaABCC3 alone had little or no effect. Inheritance of resistance was autosomal and recessive. Bioassays of progeny from interstrain crosses revealed that one wild type allele of either HaABCC2 or HaABCC3 is sufficient to sustain substantial susceptibility to Cry1Ac. In contrast with previous results, susceptibility to two insecticides derived from bacteria other than Bt (abamectin and spinetoram), was not affected by knocking out HaABCC2, HaABCC3, or both. The results here provide the first evidence that either HaABCC2 or HaABCC3 protein is sufficient to confer substantial susceptibility to Cry1Ac. The functional redundancy of these two proteins in toxicity of Cry1Ac to H. armigera is expected to reduce the likelihood of field-evolved resistance relative to disruption of a toxic process where mutations affecting a single protein can confer resistance.

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Section: Plos Pathogensmentioning

confidence: 99%

Functional redundancy of two ABC transporter proteins in mediating toxicity of Bacillus thuringiensis to cotton bollworm

Wang

Zhao

et al. 2020

PLoS Pathog

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“…Insects 2020 , 11, x 3 of 16 pyrethroids, carbamates and organophosphates, diamides and Bt proteins (e.g., Cry1F), respectively [28][29][30].…”

Section: Insect Collectionmentioning

confidence: 99%

“…Primer pairs were designed with Assay Design Software (QIAGEN, Hilden, Germany), according to sequences deposited at the National Center for Biotechnology Information (NCBI) for FAW para-type VGSC (GenBank KC435025) and ace-1 (GenBank KC435023). Primers targeting FAW RyR (GenBank MK226188) and ABCC2 (GenBank KY489760) were described elsewhere [28,30], as indicated in Table S2.…”

Section: Target-site Resistance Diagnostics By Pyrosequencingmentioning

confidence: 99%

“…Insecticide resistance is usually conferred by the insensitivity of the target receptor and/or pharmacokinetic processes modifying the rate or the properties of the insecticides delivered to the target site [27]. Amino acid substitutions/indels at the VGSC (T929I, L932F, and L1014F), AChE (A201S, G227A, and F290V), RyR (I4790M and G4946E) and ATP-binding cassette subfamily C2 transporter (ABCC2) (GC insertion and GY deletion) have been linked to resistance in S. frugiperda to pyrethroids, carbamates and organophosphates, diamides and Bt proteins (e.g., Cry1F), respectively [28][29][30].…”

Section: Introductionmentioning

confidence: 99%

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Monitoring of Target-Site Mutations Conferring Insecticide Resistance in Spodoptera frugiperda

Boaventura

Martin

Pozzebon

et al. 2020

Insects

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Fall armyworm (FAW), Spodoptera frugiperda, a major pest of corn and native to the Americas, recently invaded (sub)tropical regions worldwide. The intensive use of insecticides and the high adoption of crops expressing Bacillus thuringiensis (Bt) proteins has led to many cases of resistance. Target-site mutations are among the main mechanisms of resistance and monitoring their frequency is of great value for insecticide resistance management. Pyrosequencing and PCR-based allelic discrimination assays were developed and used to genotype target-site resistance alleles in 34 FAW populations from different continents. The diagnostic methods revealed a high frequency of mutations in acetylcholinesterase, conferring resistance to organophosphates and carbamates. In voltage-gated sodium channels targeted by pyrethroids, only one population from Indonesia showed a mutation. No mutations were detected in the ryanodine receptor, suggesting susceptibility to diamides. Indels in the ATP-binding cassette transporter C2 associated with Bt-resistance were observed in samples collected in Puerto Rico and Brazil. Additionally, we analyzed all samples for the presence of markers associated with two sympatric FAW host plant strains. The molecular methods established show robust results in FAW samples collected across a broad geographical range and can be used to support decisions for sustainable FAW control and applied resistance management.

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“…One of the major mechanisms of resistance to Cry toxin is reduced toxin binding to their specific larval midgut receptors through the disruption of the receptor genes [21]. Since the disruption of the ABC transporter subfamily C2 (ABCC2) gene was first identified to confer Cry1Ac resistance in Heliothis virescens [22], mutations of the homologous ABCC2 genes associated with Cry1A and/or Cry1F resistance have been found in several lepidopteran insects, including Plutella xylostella, Trichoplusia ni [23], Bombyx mori [24], Helicoverpa armigera [25], Spodoptera exigua [26], and Spodoptera frugiperda [27][28][29]. Recently, the CRISPR/Cas9 system has been applied to investigate the in vivo role of insect ABCC2 in the mode of action and resistance mechanisms of Bt toxins.…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Mediated Knockout of the ABCC2 Gene in Ostrinia furnacalis Confers High-Level Resistance to the Bacillus thuringiensis Cry1Fa Toxin

Wang

Huang

et al. 2020

Toxins

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The adoption of transgenic crops expressing Bacillus thuringiensis (Bt) insecticidal crystalline (Cry) proteins has reduced insecticide application, increased yields, and contributed to food safety worldwide. However, the efficacy of transgenic Bt crops is put at risk by the adaptive resistance evolution of target pests. Previous studies indicate that resistance to Bacillus thuringiensis Cry1A and Cry1F toxins was genetically linked with mutations of ATP-binding cassette (ABC) transporter subfamily C gene ABCC2 in at least seven lepidopteran insects. Several strains selected in the laboratory of the Asian corn borer, Ostrinia furnacalis, a destructive pest of corn in Asian Western Pacific countries, developed high levels of resistance to Cry1A and Cry1F toxins. The causality between the O. furnacalis ABCC2 (OfABCC2) gene and resistance to Cry1A and Cry1F toxins remains unknown. Here, we successfully generated a homozygous strain (OfC2-KO) of O. furnacalis with an 8-bp deletion mutation of ABCC2 by the CRISPR/Cas9 approach. The 8-bp deletion mutation results in a frame shift in the open reading frame of transcripts, which produced a predicted protein truncated in the TM4-TM5 loop region. The knockout strain OfC2-KO showed much more than a 300-fold resistance to Cry1Fa, and low levels of resistance to Cry1Ab and Cry1Ac (<10-fold), but no significant effects on the toxicities of Cry1Aa and two chemical insecticides (abamectin and chlorantraniliprole), compared to the background NJ-S strain. Furthermore, we found that the Cry1Fa resistance was autosomal, recessive, and significantly linked with the 8-bp deletion mutation of OfABCC2 in the OfC2-KO strain. In conclusion, in vivo functional investigation demonstrates the causality of the OfABCC2 truncating mutation with high-level resistance to the Cry1Fa toxin in O. furnacalis. Our results suggest that the OfABCC2 protein might be a functional receptor for Cry1Fa and reinforces the association of this gene to the mode of action of the Cry1Fa toxin.

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Molecular characterization of Cry1F resistance in fall armyworm, Spodoptera frugiperda from Brazil

Cited by 72 publications

References 60 publications

Functional redundancy of two ABC transporter proteins in mediating toxicity of Bacillus thuringiensis to cotton bollworm

Functional redundancy of two ABC transporter proteins in mediating toxicity of Bacillus thuringiensis to cotton bollworm

Monitoring of Target-Site Mutations Conferring Insecticide Resistance in Spodoptera frugiperda

CRISPR-Mediated Knockout of the ABCC2 Gene in Ostrinia furnacalis Confers High-Level Resistance to the Bacillus thuringiensis Cry1Fa Toxin

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