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

Wang, Jing; Ma, Huanhuan; Zhao, Shan; Huang, Jianlei; Yang, Yihua; Tabashnik, Bruce E.; Wu, Yi

doi:10.1371/journal.ppat.1008427

Cited by 67 publications

(61 citation statements)

References 50 publications

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“…The characterization of the inheritance of Bt resistance will provide important information for evaluating the risks of evolution of resistance and will make it possible to formulate effective resistance management strategies. Based on previous reports, resistance to Cry1-type toxins mediated by ABCC2 mutations was recessive or incompletely recessive [22][23][24]27,28,30,32,33]. Consistent with these results, both the high-level resistance to Cry1Fa (>300-fold) and low-level resistance to Cry1Ac (~8-fold) were inherited as a recessive mode in the knockout OfC2-KO strain of O. furnacalis.…”

Section: Discussionsupporting

confidence: 87%

“…The CRISPR-mediated knockout of P. xylostella ABCC2 conferred high levels of resistance to Cry1Aa, Cry1Ab, and Cry1Ac [30]. The double knockout of ABCC2 and ABCC3 of H. armigera confers a >15,000-fold resistance to Cry1Ac [33]. A point mutation in the ABCC2 of B. mori resulted in high levels of resistance to Cry1Ab and Cry1Ac, but not to Cry1Aa [24].…”

Section: Discussionmentioning

confidence: 99%

“…The causal relationship between ABCC2 knockout and Cry1A/Cry1F resistance has been confirmed in P. xylostella [30], S. frugiperda [31], and S. exigua [32]. Interestingly, the knockout of either ABCC2 or ABCC3 of H. armigera did not confer Cry1Ac resistance, whereas the knockout of ABCC2 and ABCC3 together resulted in extremely high levels of resistance to Cry1Ac [33]. However, until now, whether or not the ABCC2 gene of O. furnacalis (OfABCC2) is involved in Bt resistance development remains unknown.…”

Section: Introductionmentioning

confidence: 93%

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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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Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 93%

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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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“…Therefore, PxABCC2 and PxABCC3 appear to function as redundant Cry1Ac receptors for determining susceptibility of Cry1Ac in P. xylostella. This finding supports recent research by Wang et al [25] demonstrating that the knockout of ABCC2 and ABCC3 are required for Cry1Ac resistance in H. armigera.…”

Section: Plos Pathogenssupporting

confidence: 92%

“…Mutations in BmABCC3 have not been reported, however, only rescue of one receptor may be required for function. CRISPR/Cas9mediated double knockout of H. armigera ABCC2 and ABCC3 confers a >15,000-fold resistance to Cry1Ac [25]. Knockout of ABCC2 in Ostrinia furnacalis causes fairly high level of resistance to Cry1F, but not Cry1A toxins [28].…”

Section: Plos Pathogensmentioning

confidence: 99%

Resistance to Bacillus thuringiensis Cry1Ac toxin requires mutations in two Plutella xylostella ATP-binding cassette transporter paralogs

Xiang

et al. 2020

PLoS Pathog

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The diamondback moth, Plutella xylostella, is a cosmopolitan pest and the first species to develop field resistance to toxins from the gram-positive bacterium Bacillus thuringiensis (Bt). Although previous work has suggested that mutations of ATP-binding cassette transporter subfamily C2 (ABCC2) or C3 (ABCC3) genes can confer Cry1Ac resistance, here we reveal that P. xylostella requires combined mutations in both PxABCC2 and PxABCC3 to achieve high-level Cry1Ac resistance, rather than simply a mutation of either gene. We identified natural mutations of PxABCC2 and PxABCC3 that concurrently occurred in a Cry1Acresistant strain (Cry1S1000) of P. xylostella, with a mutation (R A2) causing the mis-splicing of PxABCC2 and another mutation (R A3) leading to the premature termination of PxABCC3. Genetic linkage analysis showed that R A2 and R A3 were tightly linked to Cry1Ac resistance. Introgression of R A2 and R A3 enabled a susceptible strain (G88) of P. xylostella to obtain high resistance to Cry1Ac, confirming that these genes confer resistance. To further support the role of PxABCC2 and PxABCC3 in Cry1Ac resistance, frameshift mutations were introduced into PxABCC2 and PxABCC3 singly and in combination in the G88 strain with CRISPR/Cas9 mediated mutagenesis. Bioassays of CRISPR-based mutant strains, plus genetic complementation tests, demonstrated that the deletion of PxABCC2 or PxABCC3 alone provided < 4-fold tolerance to Cry1Ac, while disruption of both genes together conferred >8,000-fold resistance to Cry1Ac, suggesting the redundant/complementary roles of PxABCC2 and PxABCC3. This work advances our understanding of Bt resistance in P. xylostella by demonstrating mutations within both PxABCC2 and PxABCC3 genes are required for high-level Cry1Ac resistance.

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ABC transporter mutations in Cry1F‐resistant fall armyworm (Spodoptera frugiperda) do not result in altered susceptibility to selected small molecule pesticides

Abdelgaffar

Perera

Jurat-Fuentes

2020

Pest Management Science

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BACKGROUND Transgenic crops producing Cry and Vip3 insecticidal proteins from the bacterium Bacillus thuringiensis provide effective control of the fall armyworm, Spodoptera frugiperda J. E. Smith. However, cases of practical S. frugiperda resistance to transgenic corn producing Cry1F, Cry1Ab and Cry1A.105 proteins have been reported in the Western hemisphere. Importantly, S. frugiperda resistance to Cry1F corn in Puerto Rico was previously associated with lower susceptibility to synthetic pesticides. When characterized, resistance to transgenic corn in S. frugiperda involved alterations in an ABC transporter subfamily C2 (SfABCC2) gene. The main goal of this work was to test the role of mutations in SfABCC2 that result in resistance to Cry1F in susceptibility to synthetic and semisynthetic small molecule pesticides. RESULTS Marginal but significantly reduced susceptibility to bifenthrin and increased susceptibility to spinetoram was detected in a Cry1F‐resitant S. frugiperda strain from Puerto Rico carrying a frameshift mutation in the SfABCC2 gene. Gene editing by CRISPR/Cas9 created a SfABCC2 knockout in a laboratory reference S. frugiperda strain. When compared to the parental reference, the knockout strain displayed 25‐fold resistance to Cry1F but no alteration in susceptibility to small molecule pesticides. CONCLUSION These results support that resistance to Cry1F due to mutations in the SfABCC2 gene do not affect susceptibility to the tested small molecule pesticides.

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

Cited by 67 publications

References 50 publications

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

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

Resistance to Bacillus thuringiensis Cry1Ac toxin requires mutations in two Plutella xylostella ATP-binding cassette transporter paralogs

ABC transporter mutations in Cry1F‐resistant fall armyworm (Spodoptera frugiperda) do not result in altered susceptibility to selected small molecule pesticides

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