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

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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.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

cv:

Cultivar

PCR:

Polymerase chain reaction

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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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Authors and Affiliations

  1. Departamento de Genética, Instituto de Biociências, Universidade Federal do Rio Grande do Sul, CP 15053, Porto Alegre, RS, 91501-970, Brazil

    Beatriz Wiebke-Strohm, Marina Borges Osorio, Lauro Bücker-Neto, Luciane Maria Pereira Passaglia, Marta Bencke, Milena Schenkel Homrich, Márcia Margis-Pinheiro & Maria Helena Bodanese-Zanettini

  2. Universidade do Vale do Rio dos Sinos, São Leopoldo, RS, Brazil

    Beatriz Wiebke-Strohm, Annette Droste, Lauro Bücker-Neto & Marta Bencke

  3. Centro Universitário Feevale, Novo Hamburgo, RS, Brazil

    Annette Droste

  4. Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil

    Giancarlo Pasquali

  5. Universidade Luterana do Brasil, Canoas, RS, Brazil

    Maria Helena Bodanese-Zanettini

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  1. Beatriz Wiebke-Strohm
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  2. Annette Droste
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  3. Giancarlo Pasquali
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  5. Lauro Bücker-Neto
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  6. Luciane Maria Pereira Passaglia
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  8. Milena Schenkel Homrich
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  9. Márcia Margis-Pinheiro
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  10. Maria Helena Bodanese-Zanettini
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Correspondence to Maria Helena Bodanese-Zanettini.

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Wiebke-Strohm, B., Droste, A., Pasquali, G. et al. Transgenic fertile soybean plants derived from somatic embryos transformed via the combined DNA-free particle bombardment and Agrobacterium system. Euphytica 177, 343–354 (2011). https://doi.org/10.1007/s10681-010-0249-1

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