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Euphytica

, Volume 177, Issue 3, pp 343–354 | Cite as

Transgenic fertile soybean plants derived from somatic embryos transformed via the combined DNA-free particle bombardment and Agrobacterium system

  • Beatriz Wiebke-Strohm
  • Annette Droste
  • Giancarlo Pasquali
  • Marina Borges Osorio
  • Lauro Bücker-Neto
  • Luciane Maria Pereira Passaglia
  • Marta Bencke
  • Milena Schenkel Homrich
  • Márcia Margis-Pinheiro
  • Maria Helena Bodanese-ZanettiniEmail author
Article

Abstract

An Agrobacterium-mediated transformation procedure for soybean [Glycine max L. Merrill] proliferating somatic embryos is here described. The Agrobacterium tumefaciens LBA4404 strain harboring pTOK233, pCAMBIA1390-olp or pH7WG2Dwrky plasmids was used to mediate gene transfer into the plant genome. Prior to Agrobacterium inoculation, proliferative soybean embryogenic clusters were microwounded by DNA-free tungsten particle bombardment. Three independent transformation experiments were performed. In Experiment I, 26 transgenic plants were obtained from a unique clone of cv Bragg, while 580 plants were recovered from 105 clones of cv IAS5. In Experiment II, a single hygromycin-resistant clone of cv BRSMG68 Vencedora was recovered and gave rise to five plants. In Experiment III, 19 plants of cv Bragg and 48 plants of IAS5 were recovered, representing five and 14 independent transformation events, respectively. PCR and Southern analyses confirmed the transgenes’ integration into plant genomes. Transgenic plants were fertile. They flowered, set pods and seeds. Transgene segregation in two T1 progenies fits the Mendelian pattern (3:1 transgenic:non-transgenic plants). This is the first report of transgenic fertile soybean plants obtained from somatic embryogenic tissues transformed by the system that combines DNA-free particle bombardment and Agrobacterium.

Keywords

Agrobacterium tumefaciens Embryogenic tissues Genetic transformation Glycine max Somatic embryogenesis 

Abbreviations

2,4-D

2,4-Dichlorophenoxyacetic acid

cv

Cultivar

PCR

Polymerase chain reaction

Notes

Acknowledgments

We would like to thank Dr. Hiei for providing the vector pTOK233. Financial support was provided by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS, Brazil), Universidade do Vale do Rio dos Sinos (UNISINOS, Brazil), Centro do Agronegócio—Casa Rural (Brazil) and BIOTECSUR Project (EU 127119).

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Beatriz Wiebke-Strohm
    • 1
    • 2
  • Annette Droste
    • 2
    • 3
  • Giancarlo Pasquali
    • 4
  • Marina Borges Osorio
    • 1
  • Lauro Bücker-Neto
    • 1
    • 2
  • Luciane Maria Pereira Passaglia
    • 1
  • Marta Bencke
    • 1
    • 2
  • Milena Schenkel Homrich
    • 1
  • Márcia Margis-Pinheiro
    • 1
  • Maria Helena Bodanese-Zanettini
    • 1
    • 5
    Email author
  1. 1.Departamento de Genética, Instituto de BiociênciasUniversidade Federal do Rio Grande do SulPorto AlegreBrazil
  2. 2.Universidade do Vale do Rio dos SinosSão LeopoldoBrazil
  3. 3.Centro Universitário FeevaleNovo HamburgoBrazil
  4. 4.Centro de BiotecnologiaUniversidade Federal do Rio Grande do SulPorto AlegreBrazil
  5. 5.Universidade Luterana do BrasilCanoasBrazil

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