Abstract
Beet curly top virus (BCTV), the causal agent of curly top disease, is an important viral pathogen of sugar beets in the western United States. BCTV is exclusively transmitted by the beet leafhopper (BLH, Neoalitarus tenellus (Baker), also referred to as Circulifer tenullus) in a circulative, non-propagative manner and is primarily managed through insecticides targeting vector. Vector-borne plant viruses have been shown to modify vector biology, behavior, or both, thereby favoring their transmission. In this study, viruliferous BLHs demonstrated increased survival compared to non-viruliferous insects when exposed to a neonicotinoid insecticide at a sub-lethal rate of 0.1x rate but not at the 1x rate. In contrast, no significant difference in survival was observed between viruliferous and non-viruliferous insects treated with a pyrethroid insecticide at either 0.1x or 1x rates. Transcriptomic analysis of BLHs exposed to a 0.1x neonicotinoid treatment revealed that differentially expressed transcripts (DETs) associated with detoxification, ubiquitination, and proteolysis, innate immunity, and stress were upregulated in viruliferous BLHs, potentially contributing to their enhanced survival. In contrast, DETs associated with locomotion and cuticular structure were concomitantly downregulated, suggesting that virus-mediated behavioral adaptations are unlikely to contribute to neonicotinoid tolerance in BLHs. This study represents the first comprehensive transcriptomic investigation of virus-mediated tolerance to a neonicotinoid insecticide.
Data availability
Data is provided within the manuscript or supplementary information files.
References
Panella, L. et al. Sugarbeet. In Yield Gains in Major U.S. Field Crops 357–395 (Wiley, 2014). https://doi.org/10.2135/cssaspecpub33.c13.
Bennett, C. W. The Curly Top Disease of Sugarbeet and Other Plants, Monograph No. 7 (The American Phytopathological Society, 1971). https://doi.org/10.1094/9780890546260.
Chen, L.-F. & Gilbertson, R. L. Chapter 17: Transmission of Curtoviruses (Beet curly top virus) by the Beet Leafhopper (Circulifer tenellus). In Vector-Mediated Transmission of Plant Pathogens (eds Li, F. & Robert, L. G.) 243–262 (The American Phytopathological Society, 2016). https://doi.org/10.1094/9780890545355.017.
Han, J. et al. Beet curly top virus affects vector biology: The first transcriptome analysis of the beet leafhopper. J. Gen. Virol. 105, 002012 (2024).
Soto, M. J. & Gilbertson, R. L. Distribution and rate of movement of the Curtovirus beet mild curly top virus (Family Geminiviridae) in the beet leafhopper. Phytopathology® 93, 478–484 (2003).
Creamer, R. Chapter 37—Beet curly top virus transmission, epidemiology, and management. In Applied Plant Virology (ed. Awasthi, L. P.) 521–527 (Academic Press, 2020). https://doi.org/10.1016/B978-0-12-818654-1.00037-2.
Strausbaugh, C. A., Wenninger, E. J. & Eujayl, I. A. Management of severe curly top in sugar beet with insecticides. Plant Dis. 96, 1159–1164 (2012).
Bass, C., Denholm, I., Williamson, M. S. & Nauen, R. The global status of insect resistance to neonicotinoid insecticides. Pestic. Biochem. Physiol. 121, 78–87 (2015).
Mauck, K., Bosque-Pérez, N. A., Eigenbrode, S. D., De Moraes, C. M. & Mescher, M. C. Transmission mechanisms shape pathogen effects on host-vector interactions: Evidence from plant viruses. Funct. Ecol. 26, 1162–1175 (2012).
Blanc, S. & Michalakis, Y. Manipulation of hosts and vectors by plant viruses and impact of the environment. Curr. Opin. Insect Sci. 16, 36–43 (2016).
Eigenbrode, S. D., Bosque-Pérez, N. A. & Davis, T. S. Insect-borne plant pathogens and their vectors: Ecology, evolution, and complex interactions. Annu. Rev. Entomol. 63, 169–191 (2018).
Severin, H. H. P. Longevity, or life histories, of leafhopper species on virus-infected and on healthy plants. Hilgardia 17, 121–137 (1946).
Lee, H. et al. Insect vector manipulation by a plant virus and simulation modeling of its potential impact on crop infection. Sci. Rep. 12, 8429 (2022).
Yan, M. et al. Molecular basis of mutual benefits between Cucurbit chlorotic yellows virus (CCYV) transmission and imidacloprid resistance in Bemisia tabaci. J. Pest Sci. 96, 489–497 (2023).
Liu, B., Preisser, E. L., Jiao, X. & Zhang, Y. Tomato yellow leaf curl virus infection alters Bemisia tabaci MED (Hemiptera: Aleyrodidae) vulnerability to flupyradifurone. J. Econ. Entomol. 113, 1922–1926 (2020).
Zhang, J. et al. Tomato Chlorosis Virus (ToCV) infection induced the resistance of Bemisia tabaci to two insecticides: Pyrethroids and flupyradifurone. Horticulturae 9, 68 (2023).
Kshatriya, K. & Gershenzon, J. Disarming the defenses: Insect detoxification of plant defense-related specialized metabolites. Curr. Opin. Plant Biol. 81, 102577 (2024).
Bass, C. et al. The evolution of insecticide resistance in the peach potato aphid, Myzus persicae. Insect Biochem. Mol. Biol. 51, 41–51 (2014).
Needham, P. H. & Sawicki, R. M. Diagnosis of resistance to organophosphorus insecticides in Myzus persicae (Sulz.). Nature 230, 125–126 (1971).
Field, L. M., Devonshire, A. L. & Forde, B. G. Molecular evidence that insecticide resistance in peach-potato aphids (Myzus persicae Sulz.) results from amplification of an esterase gene. Biochem. J. 251, 309–312 (1988).
Devonshire, A. L. & Moores, G. D. A carboxylesterase with broad substrate specificity causes organophosphorus, carbamate and pyrethroid resistance in peach-potato aphids (Myzus persicae). Pestic. Biochem. Physiol. 18, 235–246 (1982).
Field, L. M. & Foster, S. P. Amplified esterase genes and their relationship with other insecticide resistance mechanisms in English field populations of the aphid, Myzus persicae (Sulzer). Pest Manag. Sci. 58, 889–894 (2002).
Hu, J. et al. mediated thiamethoxam and clothianidin resistance is accompanied by fitness cost in the whitefly. Pest Manag. Sci. 80, 910–921 (2024).
Jin, R. et al. Overexpression of CYP6ER1 associated with clothianidin resistance in Nilaparvata lugens (Stål). Pestic. Biochem. Physiol. 154, 39–45 (2019).
Mota-Sanchez, D., Hollingworth, R. M., Grafius, E. J. & Moyer, D. D. Resistance and cross-resistance to neonicotinoid insecticides and spinosad in the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). Pest Manag. Sci. 62, 30–37 (2006).
Zhang, C., Du, S., Liu, R. & Dai, W. Overexpression of multiple cytochrome P450 genes conferring clothianidin resistance in Bradysia odoriphaga. J. Agric. Food Chem. 70, 7636–7643 (2022).
Yao, J., Zhu, Y. C. & Adamczyk, J. Responses of honey bees to lethal and sublethal doses of formulated clothianidin alone and mixtures. J. Econ. Entomol. 111, 1517–1525 (2018).
Schiering, C. et al. Feedback control of AHR signalling regulates intestinal immunity. Nature 542, 242–245 (2017).
Zeng, T. et al. AANAT1 regulates insect midgut detoxification through the ROS/CncC pathway. Commun. Biol. 7, 1–12 (2024).
Dassa, E. & Bouige, P. The ABC of ABCs: A phylogenetic and functional classification of ABC systems in living organisms. Res. Microbiol. 152, 211–229 (2001).
Sun, H., Pu, J., Chen, F., Wang, J. & Han, Z. Multiple ATP-binding cassette transporters are involved in insecticide resistance in the small brown planthopper, Laodelphax striatellus. Insect Mol. Biol. 26, 343–355 (2017).
Wang, Z. et al. Identification of candidate ATP-binding cassette transporter gene family members in Diaphorina citri (Hemiptera: Psyllidae) via adult tissues transcriptome analysis. Sci. Rep. 9, 15842 (2019).
Wilkins, R. M. Insecticide resistance and intracellular proteases. Pest Manag. Sci. 73, 2403–2412 (2017).
Cheng, L. et al. Proteomic analysis of ubiquitinated proteins from deltamethrin-resistant and susceptible strains of the diamondback moth, Plutella Xylostella L. Arch. Insect Biochem. Physiol. 90, 70–88 (2015).
Ciechanover, A., Orian, A. & Schwartz, A. L. Ubiquitin-mediated proteolysis: Biological regulation via destruction. BioEssays 22, 442–451 (2000).
Araújo, R. A., Guedes, R. N. C., Oliveira, M. G. A. & Ferreira, G. H. Enhanced proteolytic and cellulolytic activity in insecticide-resistant strains of the maize weevil, Sitophilus zeamais. J. Stored Prod. Res. 44, 354–359 (2008).
Vontas, J. et al. Gene expression in insecticide resistant and susceptible Anopheles gambiae strains constitutively or after insecticide exposure. Insect Mol. Biol. 14, 509–521 (2005).
Qiu, X. et al. Genome-wide analysis of genes associated with moderate and high DDT resistance in Drosophila melanogaster. Pest Manag. Sci. 69, 930–937 (2013).
Silva, L. B., Reis, A. P., Pereira, E. J. G., Oliveira, M. G. A. & Guedes, R. N. C. Partial purification and characterization of trypsin-like proteinases from insecticide-resistant and -susceptible strains of the maize weevil, Sitophilus zeamais. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 155, 12–19 (2010).
Livneh, I., Cohen-Kaplan, V., Cohen-Rosenzweig, C., Avni, N. & Ciechanover, A. The life cycle of the 26S proteasome: From birth, through regulation and function, and onto its death. Cell Res. 26, 869–885 (2016).
Collins, G. A. & Goldberg, A. L. The logic of the 26S proteasome. Cell 169, 792–806 (2017).
Zhao, J., Zhai, B., Gygi, S. P. & Goldberg, A. L. mTOR inhibition activates overall protein degradation by the ubiquitin proteasome system as well as by autophagy. Proc. Natl. Acad. Sci. U. S. A. 112, 15790–15797 (2015).
Saikhedkar, N., Summanwar, A., Joshi, R. & Giri, A. Cathepsins of lepidopteran insects: Aspects and prospects. Insect Biochem. Mol. Biol. 64, 51–59 (2015).
Gui, Z. Z. et al. Functional role of aspartic proteinase cathepsin D in insect metamorphosis. BMC Dev. Biol. 6, 49 (2006).
Wu, F.-Y. et al. The influence of challenge on cathepsin B and D expression patterns in the silkworm Bombyx mori L. Int. J. Ind. Entomol. Biomater. 23, 129–135 (2011).
Wu, H. W. et al. Culex pipiens pallens: Identification of genes differentially expressed in deltamethrin-resistant and -susceptible strains. Pestic. Biochem. Physiol. 79, 75–83 (2004).
Ding, T.-B., Li, J., Chen, E.-H., Niu, J.-Z. & Chu, D. Transcriptome profiling of the whitefly Bemisia tabaci MED in response to single infection of tomato yellow leaf curl virus, tomato chlorosis virus, and their co-infection. Front. Physiol. 10, 65 (2019).
He, H. et al. Impacts of Cucurbit Chlorotic Yellows Virus (CCYV) on biological characteristics of its vector Bemisia tabaci (Hemiptera: Aleyrodidae) MED species. J. Insect Sci. 21, 18 (2021).
Mahanta, D. K. et al. Groundnut bud necrosis virus modulates the expression of innate immune, endocytosis, and cuticle development-associated genes to circulate and propagate in its vector, Thrips palmi. Front. Microbiol. 13, 56 (2022).
Terra, W. R., Dias, R. O., Oliveira, P. L., Ferreira, C. & Venancio, T. M. Transcriptomic analyses uncover emerging roles of mucins, lysosome/secretory addressing and detoxification pathways in insect midguts. Curr. Opin. Insect Sci. 29, 34–40 (2018).
Perez-Vilar, J. & Hill, R. L. The structure and assembly of secreted mucins * 210. J. Biol. Chem. 274, 31751–31754 (1999).
Beyenbach, K. W. et al. The septate junction protein Tetraspanin 2A is critical to the structure and function of Malpighian tubules in Drosophila melanogaster. Am. J. Physiol. Cell Physiol. 318, C1107–C1122 (2020).
Dow, J. A. T. Insights into the Malpighian tubule from functional genomics. J. Exp. Biol. 212, 435–445 (2009).
Chahine, S. & O’Donnell, M. J. Interactions between detoxification mechanisms and excretion in Malpighian tubules of Drosophila melanogaster. J. Exp. Biol. 214, 462–468 (2011).
Farmer, J., Maddrell, S. H. P. & Spring, J. H. Absorption of fluid by the midgut of Rhodnius. J. Exp. Biol. 94, 301–316 (1981).
Michael Barrett, F. Absorption of fluid from the anterior midgut in Rhodnius. J. Insect Physiol. 28, 335–341 (1982).
Te Brugge, V., Ianowski, J. P. & Orchard, I. Biological activity of diuretic factors on the anterior midgut of the blood-feeding bug, Rhodnius prolixus. Gen. Comp. Endocrinol. 162, 105–112 (2009).
Kolawole, A. O. & Kolawole, A. N. Insecticides and bio-insecticides modulate the glutathione-related antioxidant defense system of cowpea storage bruchid (Callosobruchus maculatus). Int. J. Insect Sci. 6, IJIS.S18029 (2014).
Mack, L. K. & Attardo, G. M. Heat shock proteins, thermotolerance, and insecticide resistance in mosquitoes. Front. Insect Sci. 4, 65 (2024).
Yoshimi, T. et al. Activation of a stress-induced gene by insecticides in the midge, Chironomus yoshimatsui. J. Biochem. Mol. Toxicol. 16, 10–17 (2002).
Yadav, P. et al. Effect of temperature and insecticide stresses on Aedes aegypti larvae and their influence on the susceptibility of mosquitoes to dengue-2 virus. Southeast Asian J. Trop. Med. Public Health 36, 1139–1144 (2005).
Ayyanath, M.-M. et al. Gene expression during imidacloprid-induced hormesis in green peach aphid. Dose-Response 12, 480–497 (2014).
Ingham, V. A., Wagstaff, S. & Ranson, H. Transcriptomic meta-signatures identified in Anopheles gambiae populations reveal previously undetected insecticide resistance mechanisms. Nat. Commun. 9, 5282 (2018).
Si, F.-L. et al. HSP superfamily of genes in the malaria vector Anopheles sinensis: Diversity, phylogenetics and association with pyrethroid resistance. Malar. J. 18, 132 (2019).
Mack, L. K. & Attardo, G. M. Time-series analysis of transcriptomic changes due to permethrin exposure reveals that Aedes aegypti undergoes detoxification metabolism over 24 h. Sci. Rep. 13, 16564 (2023).
Zalucki, M. & Furlong, M. Behavior as a mechanism of insecticide resistance: Evaluation of the evidence. Curr. Opin. Insect Sci. 21, 19–25 (2017).
Sparks, T. C., Lockwood, J. A., Byford, R. L., Graves, J. B. & Leonard, B. R. The role of behavior in insecticide resistance. Pestic. Sci. 26, 383–399 (1989).
Mauck, K. E., De Moraes, C. M. & Mescher, M. C. Effects of pathogens on sensory-mediated interactions between plants and insect vectors. Curr. Opin. Plant Biol. 32, 53–61 (2016).
Balabanidou, V., Grigoraki, L. & Vontas, J. Insect cuticle: A critical determinant of insecticide resistance. Curr. Opin. Insect Sci. 27, 68–74 (2018).
Ahmad, M., Denholm, I. & Bromilow, R. H. Delayed cuticular penetration and enhanced metabolism of deltamethrin in pyrethroid-resistant strains of Helicoverpa armigera from China and Pakistan. Pest Manag. Sci. 62, 805–810 (2006).
Lin, Y., Jin, T., Zeng, L. & Lu, Y. Cuticular penetration of β-cypermethrin in insecticide-susceptible and resistant strains of Bactrocera dorsalis. Pestic. Biochem. Physiol. 103, 189–193 (2012).
Xu, Y. et al. CPR63 promotes pyrethroid resistance by increasing cuticle thickness in Culex pipiens pallens. Parasit. Vectors 15, 54 (2022).
Strausbaugh, C. A., Eujayl, I. A. & Wintermantel, W. M. Beet curly top virus strains associated with sugar beet in Idaho, Oregon, and a Western U.S. collection. Plant Dis. 101, 1373–1382 (2017).
Götz, S. et al. High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids Res. 36, 3420–3435 (2008).
Acknowledgments
We would like to thank Laura Newhard, Tyler Yamashita, and Darren Cockrell for their assistance with the insecticide trials.
Funding
This project was funded by Snake River Sugar Beet Alliance and Amalgamated Sugar.
Author information
Authors and Affiliations
Contributions
P.N and O.T. N conceived and designed the study. M.S. and J.W conducted the experiments and analyzed the data. J. H assisted with bioinformatics analysis. P.N. and M.S. wrote the draft. J.W., J.H., and O.T.N assisted with the editing of the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Supplementary Material 1
Supplementary Material 2
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
Reprints and permissions
About this article
Cite this article
Schmidtbauer, M., Withycombe, J., Han, J. et al. Virus-mediated changes in insect vector tolerance to a neonicotinoid insecticide.
Sci Rep (2026). https://doi.org/10.1038/s41598-026-40402-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-40402-z
Keywords
Neoalitarus tenellus
- Beet curly top virus
- Transcriptome
- Neonicotinoids
- Pyrethroids
Source: Ecology - nature.com
