Regional differences in susceptibility to spinosyn insecticides registered for Colorado potato beetle management in Canada

Scott, Ian M.; Vickruck, Jess; Hann, Sheldon; Krolikowski, Sophie; McKinley, Pamela; Stokes-Rees, Jessica; Hatten, Gabrielle; Moffat, Chandra E.

doi:10.1016/j.pestbp.2023.105459

Cited by 5 publications

(5 citation statements)

References 44 publications

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“…CYP9E2, CYP6A23 and lncRNA-1 (all up-regulated in OFP compared with CFP based on RNA-seq analysis) genes were selected for confirming RNA-seq results. In addition, transcript levels of another lncRNA gene, referred to as lncRNA-2 in this study (lncRNA-2, LOC111518075, XR_002723912.1), was also analyzed using RT-qPCR, as this particular gene was up-regulated in a CPB population from Ontario, Canada showing resistance to spinosad and two diamide insecticides [45], as RNA-seq can sometimes miss differentially expressed genes [61].…”

Section: Rt-qpcr Confirmation Of Rna-seq Resultsmentioning

confidence: 99%

“…The insecticide sensitive laboratory population (SLP) was originally collected from a potato field at the London Research and Development Centre, London, Ontario, Canada, and has been in continuous culture over 220 generations without insecticide exposure. This population has never been exposed to spinosad insecticides, and it has been used as a sensitive population in previous studies [45,47]. The two field populations were collected from potato farms in the province of Que ´bec, Canada.…”

Section: Cpb Populations and Rearing Conditionsmentioning

confidence: 99%

“…To confirm RNA-seq results, independent biological samples were used to perform RT-qPCR for four selected genes, three from the RNA-seq results and one additional lncRNA gene, a gene that was upregulated in a CPB population from Ontario, Canada showing resistance to spinosad, as well as two diamide insecticides [45]. Expression of the four genes was also analyzed in the SLP to determine whether changes in gene expression for the four target genes correlated with resistance levels.…”

Section: Cdna Synthesis and Rt-qpcr To Confirm Differentially Express...mentioning

confidence: 99%

“…In this study, our objective was to identify molecular mechanisms of spinosad resistance in CPB, which is a major pest of potato plants causing significant crop losses [44]. Spinosad has been used to control this pest for more than two decades; however, current reports of control failure due to resistance development in multiple field populations have raised great concern [14,45,46]. Although there are currently a few studies showing possible involvement of CYPs and miRNAs in spinosad resistance in CPB [29,30], there is still a large knowledge gap in understanding the molecular mechanism of resistance and the specific genes involved.…”

Section: Introductionmentioning

confidence: 99%

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Role of CYP9E2 and a long non-coding RNA gene in resistance to a spinosad insecticide in the Colorado potato beetle, Leptinotarsa decemlineata

Kaplanoglu,

Scott,

Vickruck

et al. 2024

PLoS ONE

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Spinosads are insecticides used to control insect pests, especially in organic farming where limited tools for pest management exist. However, resistance has developed to spinosads in economically important pests, including Colorado potato beetle (CPB), Leptinotarsa decemlineata. In this study, we used bioassays to determine spinosad sensitivity of two field populations of CPB, one from an organic farm exposed exclusively to spinosad and one from a conventional farm exposed to a variety of insecticides, and a reference insecticide naïve population. We found the field populations exhibited significant levels of resistance compared with the sensitive population. Then, we compared transcriptome profiles between the two field populations to identify genes associated primarily with spinosad resistance and found a cytochrome P450, CYP9E2, and a long non-coding RNA gene, lncRNA-2, were upregulated in the exclusively spinosad-exposed population. Knock-down of these two genes simultaneously in beetles of the spinosad-exposed population using RNA interference (RNAi) resulted in a significant increase in mortality when gene knock-down was followed by spinosad exposure, whereas single knock-downs of each gene produced smaller effects. In addition, knock-down of the lncRNA-2 gene individually resulted in significant reduction in CYP9E2 transcripts. Finally, in silico analysis using an RNA-RNA interaction tool revealed that CYP9E2 mRNA contains multiple binding sites for the lncRNA-2 transcript. Our results imply that CYP9E2 and lncRNA-2 jointly contribute to spinosad resistance in CPB, and lncRNA-2 is involved in regulation of CYP9E2 expression. These results provide evidence that metabolic resistance, driven by overexpression of CYP and lncRNA genes, contributes to spinosad resistance in CPB.

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Section: Rt-qpcr Confirmation Of Rna-seq Resultsmentioning

confidence: 99%

Section: Cpb Populations and Rearing Conditionsmentioning

confidence: 99%

Section: Cdna Synthesis and Rt-qpcr To Confirm Differentially Express...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Role of CYP9E2 and a long non-coding RNA gene in resistance to a spinosad insecticide in the Colorado potato beetle, Leptinotarsa decemlineata

Kaplanoglu,

Scott,

Vickruck

et al. 2024

PLoS ONE

Self Cite

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“…This has prompted the exploration of alternative avenues to regulate this pest, as well as the development of a better characterization of the molecular levers underlying this resistance [ 3 , 4 ]. Decreased susceptibility against various compounds, including chemicals associated with the neonicotinoids or the diamide class of insecticides, has been reported to various degrees in L. decemlineata [ 5 , 6 , 7 , 8 ]. Investigation into the molecular changes associated with responses against insecticides or toxins in L. decemlineata has highlighted multiple factors warranting closer characterization, including enzymes such as cytochrome P450 monoxygenases, glutathione s-transferases, glutathione synthetase and UDP-glycosyltransferases [ 9 , 10 , 11 ], as well as other targets of interest, including ATP-binding cassette transporter [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Cyantraniliprole and Thiamethoxam Exposure Changes Expression of Transcripts Associated with Small Non-Coding RNA Processing in the Colorado Potato Beetle

Bastarache,

Timani,

Ben Youssef

et al. 2024

Insects

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The Colorado potato beetle (Leptinotarsa decemlineata (Say)) can cause extensive damage to agricultural crops worldwide and is a significant insect pest. This insect is notorious for its ability to evade various strategies deployed to control its spread and is known for its relative ease in developing resistance against different insecticides. Various molecular levers are leveraged by L. decemlineata for this resistance to occur, and a complete picture of the genes involved in this process is lacking. While small non-coding RNAs, including miRNAs, are differentially expressed in insects exposed to insecticides, levels of transcript coding for proteins underlying their synthesis remain to be characterized fully. The overarching objective of this work aims to fill that gap by assessing the expression of such targets in L. decemlineata exposed to cyantraniliprole and thiamethoxam. The expression status of Ago1, Ago2, Ago3, Dcr2a, Dcr2b, Expo-5, Siwi-1 and Siwi-2 transcripts were quantified via qRT-PCR in adult L. decemlineata treated with low and high doses of these compounds for different lengths of time. Variation in Ago1 and Dcr2b expression was notably observed in L. decemlineata exposed to cyantraniliprole, while thiamethoxam exposure was associated with the modulation of Dcr2a and Siwi-1 transcript levels. The down-regulation of Ago1 expression in L. decemlineata using dsRNA, followed by cyantraniliprole treatment, was associated with a reduction in the survival of insects with reduced Ago1 transcript expression. Overall, this work presents the insecticide-mediated modulation of transcripts associated with small non-coding RNA processing and showcases Ago1 as a target to further investigate its relevance in cyantraniliprole response.

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Rapid Screening of Colorado Potato Beetle Resistance Derived from Solanum okadae

McCoy,

Fenstemaker,

MacKinley

et al. 2024

Am. J. Potato Res.

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Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say) is a major insect pest of potato and development of resistant varieties is part of a strategy for management. Wild relatives of potato are resources for genetic improvement through breeding. Interspecies crosses to introgress CPB resistance will be facilitated with rapid and inexpensive selection methods. Solanum okadae is a novel source of feeding deterrence against the beetle and was associated with production of a naturally occurring leaf-specific lactone-containing metabolite in the foliage. The Baljet assay has been used for decades in pharmaceuticals for rapid screening of lactone-containing compounds. A modified Baljet assay was developed for potato foliar tissue to rapidly screen for lactone-containing compounds in plants from the field, greenhouse, and laboratory. Herein we report the screening of potato foliage for CPB resistance with a Baljet assay validated by CPB larval feeding studies. Foliage from wild accessions of S. okadae were tested using the Baljet assay and results showed that production of the leaf-specific lactones has a large range of variation in the species. In addition, tubers tested using the Baljet assay had negligible levels of lactone-containing metabolites, confirming leaf-specific production of lactones. This inexpensive method using leaf disk screening will allow potato breeders to quickly select for potential CPB resistant germplasms and advance the breeding of sustainable crops.

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Regional differences in susceptibility to spinosyn insecticides registered for Colorado potato beetle management in Canada

Cited by 5 publications

References 44 publications

Role of CYP9E2 and a long non-coding RNA gene in resistance to a spinosad insecticide in the Colorado potato beetle, Leptinotarsa decemlineata

Role of CYP9E2 and a long non-coding RNA gene in resistance to a spinosad insecticide in the Colorado potato beetle, Leptinotarsa decemlineata

Cyantraniliprole and Thiamethoxam Exposure Changes Expression of Transcripts Associated with Small Non-Coding RNA Processing in the Colorado Potato Beetle

Rapid Screening of Colorado Potato Beetle Resistance Derived from Solanum okadae

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