Cloning, spatiotemporal expression and response to spinetoram, of the Tuta absoluta CYP9A304v2 gene

Author of the article:WANG Sheng-Yu1** XIE Jin-Gang1** YAN Guo-Rong2 LIN Ming-Hao1 WANG Xin-Hai1 WANG Sheng-Fei1 WANG Jian-Min1 LI Yu-Shan2 LIU Xiao-Ning1***

Author's Workplace：1. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830017, China; 2. Institute of Crop, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China

Key Words：Tuta absoluta; CYP9A304v2; spinetoram; spatiotemporal expression; bioinformatics; molecular docking

Abstract：[Aim] To investigate the potential role of the CYP9 subfamily gene CYP9A304v2 in the development of Tuta absoluta and the response of this gene to spinetoram, a biopesticide to which T. absoluta is becoming increasingly resistant. [Methods] Gene cloning, bioinformatics analysis, spatiotemporal expression profiling, stress induction experiments, and molecular docking were employed to characterize the sequence features and phylogenetic relationships of CYP9A304v2. In addition, expression patterns in different developmental stages and in response to spinetoram were measured, together with an evaluation of the binding affinity between the CYP9A304v2 protein and spinetoram. [Results] The open reading frame (ORF) of CYP9A304v2 was 1 596 bp in length, and encoded 531 amino acids. Phylogenetic analysis revealed that T. absoluta CYP9A304v2 belongs to the CYP6 family. Spatiotemporal expression profiling demonstrated specific high expression of CYP9A304v2 in the 4th instar larval and pupal stages, and that its expression was significantly upregulated under spinetoram stress (P < 0.05). Molecular docking analyses indicates strong binding affinity between the CYP9A304v2 protein and spinetoram, with binding free energies consistently below -9 kcal/mol. [Conclusion] The high expression of CYP9A304v2 during the 4th instar larval and pupal stages, combined with its strong binding affinity for spinetoram, suggests that this gene may play a critical role in both the larval-pupal transition and the detoxification of spinetoram. These results identify a candidate gene that could be targeted to control T. absoluta, and provide new information on insecticide resistance mechanisms in this species.