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Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP

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Abstract

Genetic transformation of the codling moth, Cydia pomonella, was accomplished through embryo microinjection with a plasmid-based piggyBac vector containing the enhanced green fluorescent protein (EGFP) gene. Sequencing of the flanking regions around the inserted construct resulted in identification of insect genomic sequences, not plasmid sequences, thus providing evidence that the piggyBac EGFP cassette had integrated into the codling moth genome. EGFP-positive moths were confirmed in the 28th and earlier generations post injection through PCR and Southern blot analyses, indicating heritability of the transgene.

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Acknowledgments

The authors give much thanks to Susan McCombs, Tom Miller, Frantisek Marec, Steve Garczynski, Meg Allen, and Al Handler for their reviews of the manuscript. We are indebted to Dr. Paul Shirk for his donation of plasmid constructs, p3E1.2 and p3E12Δtrl. We would also like to thank Dr. Al Handler for the pB[PUbnlsEGFP construct. We are very grateful to Eric Bruntjen, Jennifer Scott, Anne Kenny Chapman, Nina Barcenas, and Tina Vasile for their technical assistance. This project was supported by a United States Department of Agriculture-Agricultural Research Service National Program Staff funded postdoctoral position with additional funding from the Washington Tree Fruit Research Commission and the Washington State Commission on Pesticide Registration.

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Authors and Affiliations

  1. Washington State University, 24106 North Bunn Road, Prosser, WA, 99350, USA

    Holly J. Ferguson

  2. Yakima Agricultural Research Laboratory, USDA, ARS, 5230 Konnowac Pass Road, Wapato, WA, 98951, USA

    Lisa G. Neven

  3. Amgen, 1120 Veterans Blvd, South San Francisco, CA, 94080, USA

    Stephen T. Thibault

  4. Department of Biological Sciences, and Center for Global Health and Infectious Diseases, University of Notre Dame, Notre Dame, IN, 46556, USA

    Ahmed Mohammed & Malcolm Fraser

  5. Agricultural Genetic Engineering Research Institute, Agricultural Research Center, 9 Gammaa St., Giza, Egypt

    Ahmed Mohammed

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  1. Holly J. Ferguson
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  2. Lisa G. Neven
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  4. Ahmed Mohammed
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  5. Malcolm Fraser
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Correspondence to Holly J. Ferguson.

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Ferguson, H.J., Neven, L.G., Thibault, S.T. et al. Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP. Transgenic Res 20, 201–214 (2011). https://doi.org/10.1007/s11248-010-9391-8

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