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Planta

, Volume 223, Issue 6, pp 1219–1230 | Cite as

High-efficiency transformation of the diploid strawberry (Fragaria vesca) for functional genomics

  • Teruko Oosumi
  • Hope A. Gruszewski
  • Leslie A. Blischak
  • Aaron J. Baxter
  • Phillip A. Wadl
  • Joel L. Shuman
  • Richard E. Veilleux
  • Vladimir Shulaev
Original Article

Abstract

Fragaria vesca L., a diploid (2n=2x=14) relative of the commercial octoploid strawberry, is an attractive model for functional genomics research in Rosaceae. Its small genome size, short reproductive cycle, and facile vegetative and seed propagation make F. vesca a promising candidate for forward and reverse genetics experiments. However, the lack of a high-efficiency transformation protocol required for systematic production of thousands of T-DNA insertional mutant lines and high-throughput gene validation is a major bottleneck. We describe a new transformation procedure that uses leaf explants from newly unfolded trifoliate leaves obtained from stock plants 6–7 weeks after seed germination, co-cultivation with Agrobacterium strain GV3101, and stringent selection on MS medium containing 4 mg l−1 hygromycin. Using this protocol we achieved 100% transformation efficiency for 6 of 14 F. vesca accessions tested. Accession PI 551572 was determined to be the best candidate for a model in F. vesca functional genomics research, as it showed the greatest propensity for callus formation, transformation, shoot regeneration, ex vitro establishment, and plant growth, requiring only 14–15 weeks to complete its life cycle in different seasons in the greenhouse.

Keywords

Rosaceae T-DNA tagging Insertional mutagenesis Agrobacterium tumefaciens Woodland strawberry 

Abbreviations

BA

Benzyladenine

GFP

Green fluorescent protein

IBA

Indole-3-butyric acid

hpt

Hygromycin phosphotransferase

Kn

Kanamycin

Notes

Acknowledgements

This project was funded by a Virginia Tech ASPIRES grant. We thank Dr. Kim Hummer, Dr. Nalha Bassil, and Bruce Bartlett at the NCGR for providing seed and in vitro cultured material of F. vesca. We thank Scott Rapier for help with plant maintenance in the greenhouse. We thank Dr. Jerzy Nowak, Dr. Ron Mittler, and Jim Walke for critically reading the manuscript.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Teruko Oosumi
    • 1
  • Hope A. Gruszewski
    • 1
  • Leslie A. Blischak
    • 1
    • 2
  • Aaron J. Baxter
    • 1
    • 2
  • Phillip A. Wadl
    • 2
  • Joel L. Shuman
    • 1
    • 2
  • Richard E. Veilleux
    • 2
  • Vladimir Shulaev
    • 1
    • 2
  1. 1.Virginia Bioinformatics InstituteVirginia Polytechnic Institute and State UniversityBlacksburgUSA
  2. 2.Department of HorticultureVirginia Polytechnic Institute and State UniversityBlacksburgUSA

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