Germline transformation of the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), with a piggyBac transposon vector

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dc.authoridSchetelig, Prof. Dr. Marc F./0000-0002-9217-394X
dc.contributor.authorGenc, Hanife
dc.contributor.authorSchetelig, Marc F.
dc.contributor.authorNirmala, Xavier
dc.contributor.authorHandler, Alfred M.
dc.date.accessioned2025-01-27T21:00:06Z
dc.date.available2025-01-27T21:00:06Z
dc.date.issued2016
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractThe 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 assess life fitness parameters, including egg hatching, larval survival, larval-to-pupal survival, pupal-to-adult survival, and fertility. In three transgenic lines, survival at all biological stages was similar; overall fitness was significantly lower compared to wild-type olive flies, but similar to fitness levels previously reported for transgenic Mexican fruit flies. The studies presented here demonstrate the development of marked strains for olive fly using polyubiquitin-regulated fluorescent proteins in transformation vectors that can be stabilized for strain stability and ecological safety. This is the first successful effort to establish transgenic strains for an important agricultural pest in Turkey.
dc.description.sponsorshipScientific and Technological Research Council of Turkey (TUBITAK) [105 O 558]; Turkish Academy of Sciences [TUBA-GEBIP 2009]; USDA-NIFA-Biotechnology Risk Assessment Grants (BRAG) program [2011-39211-30769]; Emmy Noether Program of the German Research Foundation [SCHE 1833-1/1]; LOEWE Center for Insect Biotechnology & Bioresources of the MFS; NIFA [579242, 2011-39211-30769] Funding Source: Federal RePORTER
dc.description.sponsorshipThis research was supported by the Scientific and Technological Research Council of Turkey (TUBITAK; Project Grant No. 105 O 558; HG), the Turkish Academy of Sciences (Award No. TUBA-GEBIP 2009; HG), the USDA-NIFA-Biotechnology Risk Assessment Grants (BRAG) program (Award No. 2011-39211-30769; AMH), the Emmy Noether Program SCHE 1833-1/1 of the German Research Foundation (MFS), and the LOEWE Center for Insect Biotechnology & Bioresources of the MFS.
dc.identifier.doi10.3906/biy-1510-55
dc.identifier.endpage855
dc.identifier.issn1300-0152
dc.identifier.issn1303-6092
dc.identifier.issue4
dc.identifier.scopus2-s2.0-84975226307
dc.identifier.scopusqualityQ1
dc.identifier.startpage845
dc.identifier.urihttps://doi.org/10.3906/biy-1510-55
dc.identifier.urihttps://hdl.handle.net/20.500.12428/26940
dc.identifier.volume40
dc.identifier.wosWOS:000378689600014
dc.identifier.wosqualityQ3
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.language.isoen
dc.publisherTubitak Scientific & Technological Research Council Turkey
dc.relation.ispartofTurkish Journal of Biology
dc.relation.publicationcategoryinfo:eu-repo/semantics/openAccess
dc.rightsinfo:eu-repo/semantics/openAccess
dc.snmzKA_WoS_20250125
dc.subjectBactrocera oleae
dc.subjectolive fruit fly
dc.subjecttransformation
dc.subjectpiggyBac
dc.subjecttransposon vector
dc.titleGermline transformation of the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), with a piggyBac transposon vector
dc.typeArticle

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