Plant Cell Reports

, Volume 32, Issue 3, pp 329–337

Development of an efficient transformation method by Agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (Fragaria vesca) using NPTII selection

Authors

  • Christopher J. Pantazis
    • Institute for Advanced Learning and Research
  • Sarah Fisk
    • Institute for Advanced Learning and Research
  • Kerri Mills
    • Department of HorticultureVirginia Polytechnic Institute and State University
  • Barry S. Flinn
    • Institute for Advanced Learning and Research
    • Department of HorticultureVirginia Polytechnic Institute and State University
    • Department of Forest Resources and Environmental ConservationVirginia Polytechnic Institute and State University
  • Vladimir Shulaev
    • Department of Biological SciencesUniversity of North Texas
  • Richard E. Veilleux
    • Department of HorticultureVirginia Polytechnic Institute and State University
    • Institute for Advanced Learning and Research
    • Department of HorticultureVirginia Polytechnic Institute and State University
    • Department of Forest Resources and Environmental ConservationVirginia Polytechnic Institute and State University
Original Paper

DOI: 10.1007/s00299-012-1366-1

Cite this article as:
Pantazis, C.J., Fisk, S., Mills, K. et al. Plant Cell Rep (2013) 32: 329. doi:10.1007/s00299-012-1366-1

Abstract

Key message

We developed an efficientAgrobacterium-mediated transformation method using anAc/Dstransposon tagging construct forF. vescaand high throughput paromomycin spray assay to identify its transformants for strawberry functional genomics.

Abstract

Genomic resources for Rosaceae species are now readily available, including the Fragaria vesca genome, EST sequences, markers, linkage maps, and physical maps. The Rosaceae Genomic Executive Committee has promoted strawberry as a translational genomics model due to its unique biological features and transformability for fruit trait improvement. Our overall research goal is to use functional genomic and metabolic approaches to pursue high throughput gene discovery in the diploid woodland strawberry. F. vesca offers several advantages of a fleshy fruit typical of most fruit crops, short life cycle (seed to seed in 12–16 weeks), small genome size (206 Mbb/C), small plant size, self-compatibility, and many seeds per plant. We have developed an efficient Agrobacterium tumefaciens-mediated strawberry transformation method using kanamycin selection, and high throughput paromomycin spray assay to efficiently identify transgenic strawberry plants. Using our kanamycin transformation method, we were able to produce up to 98 independent kanamycin resistant insertional mutant lines using a T-DNA construct carrying an Ac/Ds transposon Launchpad system from a single transformation experiment involving inoculation of 22 leaf explants of F. vesca accession 551572 within approx. 11 weeks (from inoculation to soil). Transgenic plants with 1–2 copies of a transgene were confirmed by Southern blot analysis. Using our paromomycin spray assay, transgenic F. vesca plants were rapidly identified within 10 days after spraying.

Keywords

Agrobacterium tumefaciens-mediated transformationWoodland strawberryKanamycin selectionParomomycin spray assay

Copyright information

© Springer-Verlag Berlin Heidelberg 2012