use of mutagenesis, genetic mapping and next generation transcriptomics to investigate insecticide resistance mechanisms利用诱变、基因映射和下一代转录组研究杀虫剂耐药性机制.pdfVIP

Use of Mutagenesis, Genetic Mapping and Next Generation Transcriptomics to Investigate Insecticide Resistance Mechanisms 1,2 3¤ 4 5 5 Predrag Kalajdzic *, Stefan Oehler , Martin Reczko , Nena Pavlidi , John Vontas , Artemis G. Hatzigeorgiou4, Charalambos Savakis2,3 1 Institute for Biological Research, University of Belgrade, Belgrade, Serbia, 2 Medical School, University of Crete, Heraklion, Greece, 3 Institute of Cellular and Developmental Biology, Biomedical Sciences Research Center ‘‘Alexander Fleming’’, Varkiza, Greece, 4 Institute of Molecular Oncology, Biomedical Sciences Research Center ‘‘Alexander Fleming’’, Varkiza, Greece, 5 Department of Biology, University of Crete, Heraklion, Greece Abstract Insecticide resistance is a worldwide problem with major impact on agriculture and human health. Understanding the underlying molecular mechanisms is crucial for the management of the phenomenon; however, this information often comes late with respect to the implementation of efficient counter-measures, particularly in the case of metabolism-based resistance mechanisms. We employed a genome-wide insertional mutagenesis screen to Drosophila melanogaster, using a Minos-based construct, and retrieved a line (MiT[w2]3R2) resistant to the neonicotinoid insecticide Imidacloprid. Biochemical and bioassay data indicated that resistance was due to increased P450 detoxification. Deep sequencing transcriptomic analysis revealed substantial over- and under-representation of 357 transcripts in the resistant line, including statistically significant changes in mixed function oxidases, peptidases and cuticular proteins. Three P450 genes (Cyp4p2, Cyp6a2 and Cyp6g1) locat