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The olive fruit fly, Bactrocera oleae, is a highly significant pest in olive growing countries, and controlling it may be enhanced by using genetically modified strains, especially for sterile insect technique programs. | Turkish Journal of Biology Turk J Biol (2016) 40: 845-855 © TÜBİTAK doi:10.3906/biy-1510-55 http://journals.tubitak.gov.tr/biology/ Research Article Germline transformation of the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), with a piggyBac transposon vector 1, 2,3 2 2 Hanife GENÇ *, Marc F. SCHETELIG , Xavier NIRMALA , Alfred M. HANDLER Department of Agricultural Biotechnology, Faculty of Agriculture, Çanakkale Onsekiz Mart University, Çanakkale, Turkey 2 Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, US Department of Agriculture, Gainesville, Florida, USA 3 Institute for Insect Biotechnology, Justus Liebig University Giessen, Giessen, Germany 1 Received: 21.10.2015 Accepted/Published Online: 07.12.2015 Final Version: 21.06.2016 Abstract: The olive fruit fly, Bactrocera oleae, is a highly significant pest in olive growing countries, and controlling it may be enhanced by using genetically modified strains, especially for sterile insect technique programs. To improve and expand this technology, piggyBac-mediated germline transformation was achieved in a laboratory-adapted wild olive fruit fly strain. A piggyBac vector was used that is marked with both green (EGFP) and red (DsRed) fluorescent protein genes, with a duplicate piggyBac 5’ terminal inverted repeat sequence inserted between the marker genes for subsequent immobilization of vectors integrated into the host genome. Five transformant G1 adults were selected based on marker gene expression, yielding an estimated minimum germline transformation frequency of approximately 1.8% per G0 adult. All transgenic lines expressed DsRed and EGFP, although DsRed was more visible and robust compared to EGFP expression, which remained stable for more than 20 generations. Marker expression and PCR analysis, including an insertion site sequence, was consistent with stable genomic insertions. This is the first study of B. oleae transformant lines to
