Abstract
In our previous study we isolated 10 bacterial species from fourth-instar larval midguts of the red imported fire ant, Solenopsis invicta. Here we report the genetic transformation and reintroduction of three species (Kluyvera cryocrescens, Serratia marcescens, and isolate 38) into the fire ant host. All three species were transformed with the plasmid vector, pZeoDsRed. High expression levels of DsRed were observed and the plasmid is maintained in these bacteria at 37°C in the absence of antibiotic selection for at least 9 days of subculturing. The transformed bacteria were successfully reintroduced into fire ant larvae and survived in the fire ant gut for at least 7 days. Upon pupal emergence, 7 days after reintroduction, transformed bacteria can still be isolated, however, most were passed out in the meconium. We further demonstrated that the engineered bacteria could be spread within the colony by feeding this meconium to naive larvae with the aid of worker fire ants.
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Acknowledgments
This work was supported by a grant from the Texas Imported Fire Ant Research and Management Project. We are grateful to Dr. John Peloquin (Department of Entomology, University of California, Riverside) for providing us with the pZeoDsRed plasmid, and to Sherry Ellison (Entomology Research Laboratory, Texas A&M University) for her valuable support in maintaining the fire ant colonies.
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Freder Medina and Haiwen Li have contributed equally to this work.
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Medina, F., Li, H., Vinson, S.B. et al. Genetic Transformation of Midgut Bacteria from the Red Imported Fire Ant (Solenopsis invicta). Curr Microbiol 58, 478–482 (2009). https://doi.org/10.1007/s00284-008-9350-2
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DOI: https://doi.org/10.1007/s00284-008-9350-2