Abstract
Insecticide resistance complicates pest management by reducing chemical effectiveness and increasing environmental pollution from higher application rates. Monitoring resistance is essential before recommending chemicals in any agro-ecological region. The present study evaluated the extent and mechanisms of insecticide resistance in the cotton leafhopper, Amrasca biguttula biguttula (Ishida) (Hemiptera: Cicadellidae), across major cotton-growing regions of Maharashtra, India over five consecutive years, from 2015–16 to 2019–20 at ICAR-Central Institute for Cotton Research (CICR) in Nagpur. The bioassay studies using IRAC protocols revealed that populations collected from the Amravati district of Maharashtra exhibited higher LC50 values for all tested insecticides, including Flonicamid 50WG, Thiamethoxam 25WG, Acetamiprid 20SP, Imidacloprid 17.8SL, Monocrotophos 36SL, Acephate 75SP, Clothianidin 50WDG, and Spiromesifen 22.9SC. The tentative discriminating doses were established based on a susceptible population of A. biguttula at 1, 0.2, 0.05, 0.01, 0.001, 0.0001, and a control of 0 ppm. Using standardized IRAC bioassay protocols, we assessed resistance levels for eight insecticides and noted significant increases in LC50 values. Notably, the A. biguttula biguttula populations from Yavatmal, Chandrapur, and Amravati exhibited critical resistance to neonicotinoids and other classes of insecticides. Biochemical assays revealed elevated activities of detoxifying enzymes, including cytochrome P450 monooxygenases, carboxylesterases, and glutathione S-transferases (GST), indicating metabolic resistance as a key mechanism. Notably, Amravati populations displayed the highest enzyme activities (e.g., GST up to 555.56 pmol/min/mg protein), correlating with intense insecticide use. These findings emphasize the need for integrated pest management (IPM) strategies, including insecticide rotation, biological controls, and resistant cotton hybrids, to mitigate insecticide resistance and minimise environmental impact. Regular monitoring of resistance and enzyme activity is essential for sustainable pest control in cotton ecosystems.
Similar content being viewed by others
Data availability
All data generated or analysed during this study are included in this published article.
References
Rajput, A., Raghuwanshi, P. S. & Chaturvedi, P. 2023, Dynamics of area, production, and productivity of cotton crop in India. Curr. Agric. Res. J. 11(2), CARJ11.2.12.
Nagaraj, S. et al. Emerging technological developments to address pest resistance in Bt cotton. J. Cotton Res. 7(1), 30 (2024).
Halder, J., Sanwal, S. K., Deb, D., Rai, A. B. & Singh, B. Mechanisms of physical and biochemical basis of resistance against leafhopper (Amrasca biguttula biguttula) in different okra (Abelmoschus esculentus) genotypes. Indian J. Agric. Sci. 86(4), 481–484 (2016).
Manivannan, A., Kanjana, D., Dharajothi, B. & Meena, B. Evaluation of resistance in cotton genotypes against leafhoppers Amrasca biguttula biguttula (Ishida) (Homoptera: Cicadellidae). Int. J. Trop. Insect Sci. 41(4), 2409–2420 (2021).
Banoth, M. et al. Biochemical analysis of metabolites in cotton (Gossypium hirsutum L.) conferring resistance to leafhopper Amrasca biguttula biguttula (Ishida). Electron. J. Plant Breed. 14(3), 965–975 (2023).
Alagarsamy, M., Amal, T. C., Karuppan, S. & Adhimoolam, K. Comparative proteomic analysis of resistant and susceptible cotton genotypes in response to leafhopper infestation. J. Proteomics 305, 105258 (2024).
Mawblei, C., Premalatha, N., Manivannan, A. & Senguttuvan, K. Characterization and screening of cotton (Gossypium hirsutum L.) germplasm for leafhopper (Amrasca biguttula biguttula (Ishida)) resistance. J. Cotton Res. 7(1), 19 (2024).
El-Sherbeni, A., Khaleid, M. S. & Abdallah, S. Effect of some insecticides alone and in combination with salicylic acid against aphid, Aphis gossypii, and white fly Bemisia tabaci on cotton field. Bull. Natl. Res. Cent. 43(1), 57 (2019).
Sagar, D. R. A. B. & Balikai, R. A. Insecticide resistance in cotton leafhopper, Amrasca biguttula biguttula (Ishida)–a review. Biochem. Cell Arch. 14(2), 283–294 (2014).
Jan, M. T., Abbas, N., Shad, S. A. & Saleem, M. A. Resistance to organophosphate, pyrethroid and biorational insecticides in populations of spotted bollworm, Earias vittella (Fabricius) (Lepidoptera: Noctuidae), in Pakistan. Crop Prot. 78, 247–252 (2015).
Saeed, R., Razaq, M., Abbas, N., Jan, M. T. & Naveed, M. Toxicity and resistance of the cotton leafhopper, Amrasca devastans (Distant) to neonicotinoid insecticides in Punjab Pakistan. Crop Prot. 93, 143–147 (2017).
Siddiqui, J. A. et al. Insights into insecticide-resistance mechanisms in invasive species: Challenges and control strategies. Front. Physiol. 13, 1112278 (2023).
Regupathy, A. & Ayyasamy, R., Insecticide resistance in sucking pests of cotton and resurgence. In: Proc. Int. Symp. Strategies for Sustainable Cotton Production: A Global Vision, Univ. Agric. Sci., Dharwad, India, 23–25 Nov., pp. 231. (2004).
Rajashekhar, M., Prabhakar Reddy, T., Chandrashekara, K. M., Rajashekar, B., Jagan Mohan Reddy, M., Ramakrishna, K., Satyanarayana, E., Shankar, A., Jahan, A., Kumar, P. & Shravika, L., Evaluation of integrated pest management module for pink bollworm, Pectinophora gossypiella (Saunders) and its economic analysis under farmer’s field conditions. Int. J. Pest Manag. 70(4), 1212–1220. (2024).
Rajashekhar, M., Rajashekar, B., Reddy, T. P., Chandrashekara, K. M., Vanisree, K., Ramakrishna, K., Sunitha, V., Shaila, O., Sathyanarayana, E., Shahanaz & Reddy, S. S., Evaluation of farmers-friendly IPM modules for the management of fall armyworm, Spodoptera frugiperda (J. E. Smith) in maize in the hot semi-arid region of India. Sci. Rep. 14(1), 7118. (2024).
Nagaraju, M. T. et al. Effect of temperature on the biological parameters of pink bollworm, Pectinophora gossypiella Saunders (Lepidoptera: Gelechiidae). Sci. Rep. 14(1), 15047 (2024).
Bhandari, S., Thakuri, L. S., Rimal, S. & Bhatta, T. Management of okra jassid (Amrasca biguttula biguttula) through the use of botanicals and chemical pesticides under field conditions in Chitwan. Nepal. J. Agric. Food Res. 10, 100403 (2022).
Thakare, V. S., Nemade, P. W., Ghosh, P. & Landge, M. K. Insecticide resistance monitoring in Amrasca biguttula biguttula (Ishida) on cotton. Ecoscan 9, 367–371 (2016).
Rekha, S., Prabhuraj, A., Hosamani, A. & Khan, H. Bioassay of insecticides against okra leafhopper Amrasca biguttula biguttula (Ishida). Int. J. Plant Prot. 10(2), 364–368 (2017).
Kaur, J. & Pathania, M. 2022, Evaluation of different IPM modules against sucking insects in Bt cotton under southwestern Punjab conditions. Agric. Res. J. 59(5).
Mehra, P., Jakhar, A., Sheoran, S. & Kumar, D. Insecticide susceptibility monitoring in leafhopper Amrasca biguttula biguttula (Ishida) on cotton. J. Environ. Ecol. 41, 1128–1134 (2023).
Kshirsagar, S. D., Satpute, N. S. & Moharil, M. P. Monitoring of insecticide resistance in cotton leafhoppers, Amrasca biguttula biguttula (Ishida). Ann. Plant Prot. Sci. 20(2), 283–286 (2012).
Mandal, D., Bhowmik, P. & Chatterjee, M. L. Effect of newer insecticides against white fly, Bemisia tabaci (Gennadius), and jassid, Amrasca biguttula biguttula (Ishida) on cotton. Pestic. Res. J. 25(2), 117–122 (2013).
Preetha, G., Stanley, J., Manoharan, T. & Kuttalam, S. Baseline toxicity of chloronicotinyls against cotton leafhopper, Amrasca biguttula biguttula, and detection of insecticide resistance. Arch. Phytopathol. Plant Prot. 47, 2095–2105 (2014).
Pate, P. P., Bhamare, V. & Narode, M. K. Monitoring of insecticidal resistance in cotton jassid, Amrasca biguttula biguttula (Ishida) of Marathwada region of Maharashtra. J. Entomol. Zool. Stud. 8, 978–983 (2020).
Halappa, B. & Patil, R. K. Detoxifying enzyme studies on cotton leafhopper, Amrasca biguttula biguttula (Ishida), resistance to neonicotinoid insecticides in field populations in Karnataka. India. J. Plant Prot. Res. 56, 346–352 (2016).
Sandhu, R. K. & Kang, B. K. Status of insecticide resistance in leafhopper, Amrasca biguttula biguttula (Ishida) on cotton. Bioscan 10(3), 1441–1444 (2015).
MahaLakshmi, M. S. & NVVSD, P. , Insecticide resistance in field population of cotton leafhopper, Amrasca devastans (Dist.) in Guntur, Andhra Pradesh, India. Int. J. Curr. Microbiol. Appl. Sci. 9(6), 3006–3011. (2020).
Malyadri, M. Monitoring of insecticide resistance in cotton leafhopper, Amrasca biguttula biguttula (Ishida) and its management. In Master’s thesis (ed. Acharya, N. G.) (Ranga Agricultural University, 2011).
Singh, S. Transgenic cotton: Its adoption, threats, and challenges ahead. J. Entomol. Zool. Stud. 6(5), 1989–1997 (2018).
Phulse, V. B. & Udikeri, S. S. Spatio-temporal dynamics of insecticide resistance in leafhoppers Amrasca biguttula biguttula (Ishida) (Homoptera: Cicadellidae) in cotton. J. Entomol. Zool. Stud. 5(6), 936–940 (2017).
Shankarganesh, K., Nagrare, V., Amutha, M., Selvi, C. & Bagyaraj, S. Effect of insecticides on susceptibility level and detoxifying enzymes in cotton leafhopper Amrasca biguttula biguttula (Ishida). Indian J. Entomol. https://doi.org/10.5544/ije.2021.329 (2022).
Daborn, P. J., Yen, J. L. & Aitken, T. H. A single amino acid mutation in the cytochrome P450 enzyme confers insecticide resistance in Drosophila. Nature 416(6880), 440–443 (2002).
De Oliveira, A. L., Almeida, J. E. & Gomes, M. D. Metabolic resistance to insecticides in cotton pests. Pest Manag. Sci. 73(5), 1034–1041 (2017).
Silva, A. X., Jander, G., Samaniego, H., Ramsey, J. S. & Figueroa, C. C. Insecticide resistance mechanisms in the green peach aphid Myzus persicae (Hemiptera: Aphididae): A transcriptomic survey. PLoS ONE 7(6), e36366 (2012).
Tabasian, H., Ravan, S., Bandani, A. R. & Siahsar, B. A. The effect of esterase activity in resistance of Aphis gossypii to selective insecticides. J. Food Agric. Environ. 8, 1108–1112 (2010).
Ullah, K. R. M., Gao, F., Sikandar, A. & Wu, H. Insights into the effects of insecticides on aphids (Hemiptera: Aphididae): Resistance mechanisms and molecular basis. Int. J. Mol. Sci. 24(7), 6750 (2023).
Finney, D. J. Probit Analysis (Cambridge University Press, 1972).
Plus, P. POLO-PC: A user’s guide to probit or logit analysis (LeOra Software, 1987).
Van Asch, B. & Van der Straten, M. J. A microplate method for the determination of esterase activity in individual insects. Pestic. Biochem. Physiol. 52(1), 1–6 (1995).
Chalam, M. S. V. & Subbaratnam, G. V. Role of mixed function oxidases in imparting resistance to the cotton leafhopper, Amrasca biguttula biguttula (Ishida). Pest Manag. Econ. Zool. 9(1), 49–53 (2001).
Acknowledgements
The first author expresses gratitude to the Director, ICAR- CICR, Nagpur for providing financial assistance, support, and reliability throughout the study.
Funding
The authors received funding through in-house funding from ICAR- CICR, Nagpur to carry out the study was duly acknowledged. The manuscript is a part of “Elucidating eco-toxicity and resistance development in sucking pests against newer insecticides used in cotton” project.
Author information
Authors and Affiliations
Contributions
V.C.B.N.: Conceptualization, Study design, Writing – original draft. L.R.C.: Formal analysis, Writing – review & editing. N.G.: Formal analysis, Writing – review & editing. N.V.S.: Review, Editing, Supervision. J.S.V.M.: Formal analysis, Writing – review & editing. G.S.S.: Critical revision of the manuscript. M.T.N.: Review, Editing. S.K.: Investigation, Data collection (conducted the experiment). Y.G.P.: Review, Editing. K.M.C.: Formal analysis, Writing – review & editing, Validation.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethical approval
Insect collection was conducted with minimal disturbance to habitats, using non-invasive sampling methods, and only the necessary number of specimens was collected to achieve statistically robust results. Collections were carried out in compliance with local regulations and guidelines for fieldwork.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
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/.
About this article
Cite this article
Chinna Babu Naik, V., Chowdary, L.R., Nagaharish, G. et al. Monitoring insecticide resistance in cotton leafhopper in relation to enzymatic activity in major cotton growing areas of central India. Sci Rep (2026). https://doi.org/10.1038/s41598-026-36055-7
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-36055-7


