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Plant Cell Reports

, Volume 29, Issue 9, pp 1013–1021 | Cite as

Backbone-free transformation of barrel medic (Medicago truncatula) with a Medicago-derived transfer DNA

  • Massimo Confalonieri
  • Roberto Borghetti
  • Anca Macovei
  • Claudia Testoni
  • Daniela Carbonera
  • Manuel Pedro Salema Fevereiro
  • Caius Rommens
  • Kathy Swords
  • Efisio Piano
  • Alma BalestrazziEmail author
Original Paper

Abstract

In the present work, Agrobacterium tumefaciens-mediated genetic transformation of the model legume Medicago truncatula Gaertn. (barrel medic) was carried out using the pSIM843 vector that contains a Medicago-derived transfer DNA, delineated by a 25-bp sequence homologous to bacterial T-DNA borders. The transfer DNA contains an expression cassette for the nptII (neomycin phosphotransferase) gene and is flanked by an expression cassette for the backbone integration marker gene ipt (isopentenyl transferase). Our results demonstrate that the Medicago-derived RB-like elements efficiently support DNA mobilization from A. tumefaciens to M. truncatula. Kanamycin-resistant shoots with normal phenotype and ipt-shooty lines were recovered at a frequency of 11.7 and 7.8%, respectively. Polymerase chain reaction (PCR) analyses demonstrated that 44.4% of the independent transgenic lines were backbone-free and evidenced the occurrence of backbone-transfer events.

Keywords

Backbone transfer ipt Medicago truncatula Morphological marker P-DNA 

Notes

Acknowledgments

This research was supported by Fondo di Ateneo per la Ricerca-University of Pavia. Authors are grateful to Dr. Cristina Ambroggi for excellent technical contribution and Dr. Lorenzo Concia for helpful suggestions concerning the interpretation of QRT-PCR data.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Massimo Confalonieri
    • 1
  • Roberto Borghetti
    • 2
  • Anca Macovei
    • 2
  • Claudia Testoni
    • 2
  • Daniela Carbonera
    • 2
  • Manuel Pedro Salema Fevereiro
    • 3
  • Caius Rommens
    • 4
  • Kathy Swords
    • 4
  • Efisio Piano
    • 1
  • Alma Balestrazzi
    • 2
    Email author
  1. 1.C.R.A.-Centro di Ricerca per le Produzioni Foraggere e Lattiero-CasearieLodiItaly
  2. 2.Dipartimento di Genetica e MicrobiologiaUniversità di PaviaPaviaItaly
  3. 3.Instituto de Tecnologia Quimica e BiológicaUniversidade Nova de LisboaOeirasPortugal
  4. 4.Simplot Plant SciencesJ.R. Simplot CompanyBoiseUSA

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