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Agrobacterium-mediated transformation of maize

Nature Protocols volume 2, pages 1614–1621 (2007)Cite this article

Abstract

Maize may be transformed very efficiently using Agrobacterium tumefaciens-mediated methods. The most critical factor in the transformation protocol is the co-cultivation of healthy immature embryos of the correct developmental stage with A. tumefaciens; the embryos should be collected only from vigorous plants grown in well-conditioned glasshouses. With the protocol described here, approximately 50% of immature embryos from the inbred line A188 and 15% from inbred lines A634, H99 and W117 will produce transformants. About half of the transformed plants are expected to carry one or two copies of the transgenes, which are inherited by the progeny in a mendelian fashion. More than 90% of transformants are expected to be normal in morphology. The protocol takes about 3 months from the start of co-cultivation to the planting of transformants into pots.

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Figure 1: Timeline of maize transformation protocol.
Figure 2: Size of immature embryo.
Figure 3: Isolation of immature embryo.
Figure 4: Guide to cutting type I callus for third selection and regeneration step.
Figure 5: Assay of segregation of transgene in detached leaves.
Figure 6: Transient expression of GUS gene in immature embryos.

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Acknowledgements

We thank Ms. E. Usami and Ms. M. Noguchi for skillful assistance.

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Authors and Affiliations

  1. Plant Innovation Center, Japan Tobacco Inc., 700 Higashibara, Iwata, 438-0802, Shizuoka, Japan

    Yuji Ishida, Yukoh Hiei & Toshihiko Komari

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  1. Yuji Ishida
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  2. Yukoh Hiei
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  3. Toshihiko Komari
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Corresponding author

Correspondence to Yuji Ishida.

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Ishida, Y., Hiei, Y. & Komari, T. Agrobacterium-mediated transformation of maize. Nat Protoc 2, 1614–1621 (2007). https://doi.org/10.1038/nprot.2007.241

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