Expressing a moth abcc2 gene in transgenic Drosophila causes susceptibility to Bt Cry1Ac without requiring a cadherin-like protein receptor |
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| Institution: | 1. Dow AgroSciences, 9330 Zionsville Rd, Indianapolis, IN 46268, United States;2. University of Nebraska-Lincoln, 103 Entomology Hall, Lincoln, NE 68583, United States;3. University of Florida, Entomology and Nematology Department, Charles Steinmetz Hall, PO Box 110620, Gainesville, FL 32611-0620, United States;1. Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan;2. Japan Society for the Promotion of Science, Japan;1. Institute of Entomology, School of Life Sciences, Central China Normal University, Wuhan 430079, China;2. Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, China;3. State Key Laboratory for Biology of Plant Disease and Insect Pests, Chinese Academy of Agricultural Sciences, West Yuanmingyuan Road, Beijing 100193, China;4. Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, Cuernavaca 62250, Morelos, Mexico;1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2. Key Laboratory of Integrated Pest Management on Crops in Central China, Ministry of Agriculture, Hubei Key Laboratory of Crop Disease, Insect Pests and Weeds Control, Institute of Plant Protection and Soil Fertility, Hubei Academy of Agricultural Sciences, Wuhan 430064, China;3. School of Life Science, Central China Normal University, Wuhan, 430079, China;4. Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, 518120, China;5. Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, Huazhong Agricultural University, Wuhan 430070, China;6. USDA, ARS, U.S. Arid Land Agricultural Research Center, Maricopa, AZ 85138, USA;7. Department of Entomology, University of Arizona, Tucson, AZ 85721, USA |
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| Abstract: | Bt toxins ingested by insect pests can bind to midgut receptors and cause death, although several steps in this process remain unclear. Multiple Bt toxin receptors have been identified in Lepidoptera, including a cadherin-like protein (CaLP), which is central to several models explaining Bt toxins’ mode of action. Mutations in the Plutella xylostella ATP-dependent binding cassette transporter C2 (Px-abcc2), rather than CaLP, are genetically linked with Bt Cry1Ac resistance. Here we expressed Px-abcc2 in Drosophila and performed larval bioassays to determine whether this protein acts as an effective Bt receptor. Cry1Ac had no effect on larvae expressing Px-abcc2 in salivary glands, yet larvae expressing Px-abcc2 in the midgut were highly susceptible to both Cry1Ac protoxin and trypsin activated toxin. Furthermore, the CaLP orthologue has been lost from the Drosophila genome, making this a useful system for investigating the role of CaLP peptides from Manduca sexta (CR12-MPED), which are known to act as Bt synergists in larval feeding assays. Drosophila larvae expressing Px-ABCC2 in the midgut were fed LD50 concentrations of Cry1Ac toxin or protoxin, plus purified CR12-MPED cloned from M. sexta or P. xylostella. The M. sexta CR12-MPED protein acted synergistically with Cry1Ac protoxin and activated toxin significantly more effectively than the P. xylostella peptide. This work demonstrates ABCC2 is the major functional Cry1Ac receptor for P. xylostella and the importance of CaLP proteins in Bt mode of action may vary between different lepidopteran species. |
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| Keywords: | Bt toxin Cry1Ac Insecticide resistance Insecticide receptor |
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