Multiple Mutations in the β1 Subunit of the Nicotinic Acetylcholine Receptor Confer Resistance to Neonicotinoids.

Journal: Journal of agricultural and food chemistry
Published Date: May 12, 2025

Abstract

The evolution of resistance to neonicotinoid insecticides threatens global agriculture. To elucidate its molecular basis, we employed as a model system to investigate resistance-associated mutations in the β1 subunit of nicotinic acetylcholine receptors (nAChRs). Using a CRISPR/Cas9-mediated allele replacement, we generated homozygous knock-in mutants (V62I, V101I, R81E, and A60T,R81E) without apparent fitness costs. Toxicity bioassays revealed that these mutations confer varying resistance levels, with the R81E mutation exhibiting over 225-fold resistance to thiamethoxam, clothianidin, and dinotefuran. A heteropentameric α1β1 nAChR model, generated using an AI-based protein-ligand prediction (Chai-1), showed that imidacloprid binds at the orthosteric site, where R81 forms a critical electrostatic interaction. Residues A60, V62, and V101, positioned further from the binding site, showed spatial distances correlated with their resistance ratios. These findings provide genetic and structural insights into neonicotinoid resistance mechanisms, offering a foundation for the design of next-generation insecticides and resistance management strategies.

Authors

  • Yang Cheng
    Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China.
  • Yu-Xin Chen
    Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China.
  • Jia-Nan Gao
    Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China.
  • Jie Chen
    School of Basic Medical Sciences, Health Science Center, Ningbo University, Ningbo, China.
  • Jia Huang
    Shanghai Lung Tumor Clinical Medical Center, Shanghai Chest Hospital, Shanghai Jiao Tong University (SJTU), Shanghai 200030, China.
  • Xiaomu Qiao
    Collaborative Innovation Center of Green Pesticide, National Joint Local Engineering Laboratory of Biopesticide Preparation, School of Forestry and Biotechnology, Zhejiang A&F University, Hangzhou 311300, China.

Keywords

No keywords available for this article.

External Resources

Popular Topics
Recent Journals