{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:55:53Z","timestamp":1785426953108,"version":"3.56.0"},"reference-count":46,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,6,7]],"date-time":"2026-06-07T00:00:00Z","timestamp":1780790400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32272576"],"award-info":[{"award-number":["32272576"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32272576"],"award-info":[{"award-number":["32272576"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"crossref","award":["KJJQ2025016"],"award-info":[{"award-number":["KJJQ2025016"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Guidance Foundation of the Sanya Institute of Nanjing Agricultural University","award":["NAUSYMS15"],"award-info":[{"award-number":["NAUSYMS15"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>The rice leaffolder, Cnaphalocrocis medinalis, has developed widespread resistance to conventional insecticides, severely threatening rice production. Pesticide combination is a critical strategy for enhancing efficacy and delaying resistance evolution. In this study, binary mixtures with distinct modes of action were screened against C. medinalis neonates. Bioassays indicated that mixtures of spinosad and indoxacarb exhibited significant synergism, while several other combinations showed antagonism. To elucidate the synergistic mechanism of the optimal 1:1 mixture, comparative transcriptomic analyses were performed. Results revealed a time-dependent dual-phase response. At 6 h, the mixture induced broader upregulation of detoxification genes and activated stress signaling pathways due to competitive enzyme saturation, impairing metabolic clearance and amplifying neurotoxicity. At 24 h, massive transcriptional reprogramming uniquely activated lysosome, autophagy, lipid metabolism, and neurodegenerative disease pathways. This study demonstrates that the synergism arises from early competitive inhibition of detoxification and late overactivation of the autophagy\u2013lysosomal system associated with cellular damage. These findings provide transcriptomic evidence for the dynamic synergistic mechanism and offer a scientific basis for rational resistance management of C. medinalis.<\/jats:p>","DOI":"10.3390\/insects17060598","type":"journal-article","created":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T07:43:02Z","timestamp":1780904582000},"page":"598","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Transcriptomic Insights into the Synergistic Mechanism of Spinosad\u2013Indoxacarb Mixtures Against Cnaphalocrocis medinalis"],"prefix":"10.3390","volume":"17","author":[{"given":"Hong-Shuang","family":"Li","sequence":"first","affiliation":[{"name":"College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"College of Plant Protection, Sanya Institute of Nanjing Agricultural University, Sanya 572025, China"},{"name":"State Key Laboratory of Agricultural and Forestry Biosecurity, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng-Zhen","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"College of Plant Protection, Sanya Institute of Nanjing Agricultural University, Sanya 572025, China"},{"name":"State Key Laboratory of Agricultural and Forestry Biosecurity, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ji-Yang","family":"Xing","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"College of Plant Protection, Sanya Institute of Nanjing Agricultural University, Sanya 572025, China"},{"name":"State Key Laboratory of Agricultural and Forestry Biosecurity, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1991-0322","authenticated-orcid":false,"given":"Cong-Fen","family":"Gao","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"College of Plant Protection, Sanya Institute of Nanjing Agricultural University, Sanya 572025, China"},{"name":"State Key Laboratory of Agricultural and Forestry Biosecurity, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0096-147X","authenticated-orcid":false,"given":"Shun-Fan","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China"},{"name":"College of Plant Protection, Sanya Institute of Nanjing Agricultural University, Sanya 572025, China"},{"name":"State Key Laboratory of Agricultural and Forestry Biosecurity, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7163","DOI":"10.1038\/s41467-021-27424-z","article-title":"Sustainable Intensification for a Larger Global Rice Bowl","volume":"12","author":"Yuan","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1038\/s41559-018-0793-y","article-title":"The Global Burden of Pathogens and Pests on Major Food Crops","volume":"3","author":"Savary","year":"2019","journal-title":"Nat. Ecol. Evol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3290","DOI":"10.1002\/ps.7512","article-title":"Insecticide Resistance Monitoring of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) and Its Mechanism to Chlorantraniliprole","volume":"79","author":"Sun","year":"2023","journal-title":"Pest Manag. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"106765","DOI":"10.1016\/j.pestbp.2025.106765","article-title":"Insecticide Resistance Monitoring and Identification of Ryanodine Receptor Mutations Driving Diamide Resistance in Cnaphalocrocis medinalis","volume":"216","author":"Li","year":"2026","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Sittichok, S., Passara, H., Sinthusiri, J., Moungthipmalai, T., Puwanard, C., Murata, K., and Soonwera, M. (2024). Synergistic Larvicidal and Pupicidal Toxicity and the Morphological Impact of the Dengue Vector (Aedes aegypti) Induced by Geranial and Trans-Cinnamaldehyde. Insects, 15.","DOI":"10.3390\/insects15090714"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"786","DOI":"10.1093\/jee\/toy376","article-title":"Effect of Synergists on Deltamethrin Resistance in the Common Bed Bug (Hemiptera: Cimicidae)","volume":"112","author":"Romero","year":"2019","journal-title":"J. Econ. Entomol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cedergreen, N. (2014). Quantifying Synergy: A Systematic Review of Mixture Toxicity Studies within Environmental Toxicology. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0096580"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Zhao, R., Li, D., Wang, X., Li, Z., Yu, X., and Shentu, X. (2022). Synergistic and Additive Interactions of Zhongshengmycin to the Chemical Insecticide Pymetrozine for Controlling Nilaparvata lugens (Hemiptera: Delphacidae). Front. Physiol., 13.","DOI":"10.3389\/fphys.2022.875610"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.aquatox.2015.12.007","article-title":"What Causes the Difference in Synergistic Potentials of Propiconazole and Prochloraz toward Pyrethroids in Daphnia magna?","volume":"172","author":"Dalhoff","year":"2016","journal-title":"Aquat. Toxicol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6974","DOI":"10.1038\/s41598-022-10812-w","article-title":"Toxicity and Biochemical Impact of Methoxyfenozide\/Spinetoram Mixture on Susceptible and Methoxyfenozide-Selected Strains of Spodoptera littoralis (Lepidoptera: Noctuidae)","volume":"12","author":"Ahmed","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3211","DOI":"10.1007\/s00204-017-2043-5","article-title":"Toxicological Interactions of Pesticide Mixtures: An Update","volume":"91","author":"Gil","year":"2017","journal-title":"Arch. Toxicol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1585","DOI":"10.1021\/acs.est.0c04493","article-title":"Can Organophosphates and Carbamates Cause Synergisms by Inhibiting Esterases Responsible for Biotransformation of Pyrethroids?","volume":"55","author":"Cao","year":"2021","journal-title":"Environ. Sci. Technol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"137023","DOI":"10.1016\/j.jhazmat.2024.137023","article-title":"Transgenerational Combined Toxicity Effects of Neonicotinoids and Triazole Pesticides at Environmentally Relevant Concentrations on D. magna: From Individual to Population Level","volume":"486","author":"Wu","year":"2025","journal-title":"J. Hazard. Mater."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.cbi.2016.06.003","article-title":"Effects of Pesticide Mixtures in Human and Animal Models: An Update of the Recent Literature","volume":"254","author":"Rizzati","year":"2016","journal-title":"Chem.-Biol. Interact."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1093\/jee\/53.5.887","article-title":"Analysis of Joint Action of Insecticides against House Flies","volume":"53","author":"Sun","year":"1960","journal-title":"J. Econ. Entomol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"106575","DOI":"10.1016\/j.pestbp.2025.106575","article-title":"Unravelling the Novel Mode of Action of the Spinosyn Insecticides: A 25 Year Review","volume":"214","author":"Perry","year":"2025","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2347","DOI":"10.1002\/ps.8628","article-title":"The Stereoselective Bioactivity and Mechanism of Indoxacarb against Spodoptera frugiperda","volume":"81","author":"Guo","year":"2025","journal-title":"Pest Manag. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1603\/ME13207","article-title":"Cross Resistances in Spinosad-Resistant Culex quinquefasciatus (Diptera: Culicidae)","volume":"51","author":"Su","year":"2014","journal-title":"J. Med. Entomol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1016\/0020-1790(75)90041-4","article-title":"Pratt Inhibition of Juvenile Hormone Carboxyesterase of Locust Haemolymph by Organophosphates in Vitro","volume":"5","author":"Grahame","year":"1975","journal-title":"Insect Biochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1146\/annurev.ento.51.110104.151104","article-title":"Molecular Mechanisms of Metabolic Resistance to Synthetic and Natural Xenobiotics","volume":"52","author":"Li","year":"2007","journal-title":"Annu. Rev. Entomol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"105707","DOI":"10.1016\/j.pestbp.2023.105707","article-title":"Regulation of CncC in Insecticide-Induced Expression of Cytochrome P450 CYP9A14 and CYP6AE11 in Helicoverpa armigera","volume":"197","author":"Wu","year":"2023","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_22","first-page":"631","article-title":"Synergism of Insecticides by Enzyme Inhibitors in the Resistant Populations of Spodoptera litura (Lepidoptera: Noctuidae)","volume":"52","author":"Ahmad","year":"2009","journal-title":"Acta Entomol. Sin."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1038\/s42003-025-07560-8","article-title":"Inhibition of Transcriptional Regulation of Detoxification Genes Contributes to Insecticide Resistance Management in Spodoptera exigua","volume":"8","author":"Hu","year":"2025","journal-title":"Commun. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/S0261-2194(00)00070-3","article-title":"Bioactivation and Mode of Action of the Oxadiazine Indoxacarb in Insects","volume":"19","author":"Wing","year":"2000","journal-title":"Crop Prot."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Flores, A.E., Davila-Barboza, J.A., and Juache-Villagrana, A.E. (2026). Insecticide Resistance and Vector Control. Trop. Med. Infect. Dis., 11.","DOI":"10.3390\/tropicalmed11010033"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1146\/annurev-ento-120811-153645","article-title":"Neuroactive Insecticides: Targets, Selectivity, Resistance, and Secondary Effects","volume":"58","author":"Casida","year":"2013","journal-title":"Annu. Rev. Entomol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"169923","DOI":"10.1016\/j.peptides.2018.02.001","article-title":"Roles of the Insulin Signaling Pathway in Insect Development and Organ Growth","volume":"122","author":"Lin","year":"2019","journal-title":"Peptides"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.molimm.2020.10.020","article-title":"Transcriptome Sequencing Reveals Cnaphalocrocis medinalis against Baculovirus Infection by Oxidative Stress","volume":"129","author":"Han","year":"2021","journal-title":"Mol. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.tig.2004.01.003","article-title":"The Genetics and Genomics of Insecticide Resistance","volume":"20","author":"Daborn","year":"2004","journal-title":"Trends Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"104727","DOI":"10.1016\/j.pestbp.2020.104727","article-title":"The Insulin Signaling Pathway in Drosophila melanogaster: A Nexus Revealing an \u201cAchilles\u2019 Heel\u201d in DDT Resistance","volume":"171","author":"Zhang","year":"2021","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"106932","DOI":"10.1016\/j.pestbp.2025.106932","article-title":"Depleting SfCarE: Innovative Strategies for Enhancing Chlorantraniliprole Toxicity through Mixing with Carbaryl in Spodoptera frugiperda (Lepidoptera: Noctuidae)","volume":"218","author":"Wang","year":"2026","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1146\/annurev-pathol-020712-163918","article-title":"Autophagy as a Stress-Response and Quality-Control Mechanism: Implications for Cell Injury and Human Disease","volume":"8","author":"Murrow","year":"2013","journal-title":"Annu. Rev. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/bs.ircmb.2020.01.003","article-title":"Crosstalk between Apoptosis and Autophagy Signaling Pathways","volume":"Volume 352","author":"Fairlie","year":"2020","journal-title":"International Review of Cell and Molecular Biology"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Eriksson, I., Ward, L.J., Vainikka, L., Sultana, N., Leanderson, P., Flodin, U., Li, W., and Yuan, X.-M. (2023). Imidacloprid Induces Lysosomal Dysfunction and Cell Death in Human Astrocytes and Fibroblasts-Environmental Implication of a Clinical Case Report. Cells, 12.","DOI":"10.3390\/cells12242772"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Ma, Y., and Wang, Q. (2025). Neonicotinoid-Induced Cytotoxicity: Insights into Cellular Mechanisms and Health Risks. Toxics, 13.","DOI":"10.3390\/toxics13070576"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.4103\/NRR.NRR-D-24-01412","article-title":"Regulation of Synaptic Function and Lipid Metabolism","volume":"21","author":"Zhang","year":"2026","journal-title":"Neural Regen. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1515\/hsz-2017-0148","article-title":"The Sphingomyelin Synthase Family: Proteins, Diseases, and Inhibitors","volume":"398","author":"Chen","year":"2017","journal-title":"Biol. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1002\/ps.5233","article-title":"Insect Midgut Structures and Molecules as Targets of Plant-Derived Protease Inhibitors and Lectins","volume":"75","author":"Albuquerque","year":"2019","journal-title":"Pest Manag. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1007\/s11356-017-0569-y","article-title":"Sublethal Dose of Deltamethrin Damage the Midgut Cells of the Mayfly Callibaetis radiatus (Ephemeroptera: Baetidae)","volume":"25","author":"Santos","year":"2018","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Feyereisen, R. (2012). Insect CYP Genes and P450 Enzymes. Insect Molecular Biology and Biochemistry, Academic Press.","DOI":"10.1016\/B978-0-12-384747-8.10008-X"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"110383","DOI":"10.1016\/j.jtbi.2020.110383","article-title":"Determinants of Optimal Insecticide Resistance Management Strategies","volume":"503","author":"Helps","year":"2020","journal-title":"J. Theor. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.jinsphys.2009.10.004","article-title":"Mechanisms of dsRNA Uptake in Insects and Potential of RNAi for Pest Control: A Review","volume":"56","author":"Huvenne","year":"2010","journal-title":"J. Insect Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"101184","DOI":"10.1016\/j.cois.2024.101184","article-title":"New and Emerging Mechanisms of Insecticide Resistance","volume":"63","author":"Pu","year":"2024","journal-title":"Curr. Opin. Insect Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"104822","DOI":"10.1016\/j.pestbp.2021.104822","article-title":"Transcriptional Regulation of Xenobiotic Detoxification Genes in Insects\u2014An Overview","volume":"174","author":"Amezian","year":"2021","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Le Goff, G., and Nauen, R. (2021). Recent Advances in the Understanding of Molecular Mechanisms of Resistance in Noctuid Pests. Insects, 12.","DOI":"10.3390\/insects12080674"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Jambagi, S., Gaddanakeri, S., Dileep, K.N.T., and Chaitra, D.S. (2023). Insecticide Resistance Management Strategies for Sustainable Insect Control. Entomology Redefined, CRC Press.","DOI":"10.1201\/9781003532569-15"}],"container-title":["Insects"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4450\/17\/6\/598\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T04:47:44Z","timestamp":1780980464000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4450\/17\/6\/598"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,7]]},"references-count":46,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["insects17060598"],"URL":"https:\/\/doi.org\/10.3390\/insects17060598","relation":{},"ISSN":["2075-4450"],"issn-type":[{"value":"2075-4450","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,7]]}}}