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Improvement of Agrobacterium-mediated transformation in Hi-II maize (Zea mays) using standard binary vectors

Plant Cell Reports volume 27, pages 297–305 (2008)Cite this article

Abstract

High-frequency transformation of maize (Zea mays L.) using standard binary vectors is advantageous for functional genomics and other genetic engineering studies. Recent advances in Agrobacterium tumefaciens-mediated transformation of maize have made it possible for the public to transform maize using standard binary vectors without a need of the superbinary vector. While maize Hi-II has been a preferred maize genotype to use in various maize transformation efforts, there is still potential and need in further improving its transformation frequency. Here we report the enhanced Agrobacterium-mediated transformation of immature zygotic embryos of maize Hi-II using standard binary vectors. This improved transformation process employs low-salt media in combined use with antioxidant l-cysteine alone or l-cysteine and dithiothreitol (DTT) during the Agrobacterium infection stage. Three levels of N6 medium salts, 10, 50, and 100%, were tested. Both 10 and 50% salts were found to enhance the T-DNA transfer in Hi-II. Addition of DTT to the cocultivation medium also improves the T-DNA transformation. About 12% overall and the highest average of 18% transformation frequencies were achieved from a large number of experiments using immature embryos grown in various seasons. The enhanced transformation protocol established here will be advantageous for maize genetic engineering studies including transformation-based functional genomics.

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Acknowledgments

We thank Regina Wamsley (from Zhanyuan J. Zhang’s laboratory) for her excellent technical assistance; CAMBIA (Australia) for providing pCAMBIA3301; Aventis CropScience (Research Triangle Park, NC, USA) for herbicide Liberty®, and James A. Birchler and Seth D. Findley (University of Missouri-Columbia) for a critical review of this manuscript. University of Missouri-Columbia Life Science Mission Enhancement program supported Angela R. Kennon and, in part, Xinlu Chen (from Zhanyuan J. Zhang’s lab). All transformation experiments were conducted in the Plant Transformation Core Facility at the University of Missouri-Columbia.

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Affiliations

  1. Departamento de Genetica, Facultad de Biologia, Universidad Complutense, 28040, Madrid, Spain

    Juan M. Vega

  2. Division of Biological Science, College of Art and Science, University of Missouri, Columbia, MO, 65211-7400, USA

    Weichang Yu

  3. Plant Transformation Core Facility, Division of Plant Sciences, College of Agriculture, Food and Natural Resources, University of Missouri, Columbia, MO, 65211-7140, USA

    Angela R. Kennon, Xinlu Chen & Zhanyuan J. Zhang

Authors
  1. Juan M. Vega
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  2. Weichang Yu
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  3. Angela R. Kennon
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  4. Xinlu Chen
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  5. Zhanyuan J. Zhang
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Corresponding author

Correspondence to Zhanyuan J. Zhang.

Additional information

Communicated by P. Ozias-Akins.

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Vega, J.M., Yu, W., Kennon, A.R. et al. Improvement of Agrobacterium-mediated transformation in Hi-II maize (Zea mays) using standard binary vectors. Plant Cell Rep 27, 297–305 (2008). https://doi.org/10.1007/s00299-007-0463-z

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  • DOI: https://doi.org/10.1007/s00299-007-0463-z

Keywords

  • Agrobacterium tumefaciens
  • Binary vector
  • Hi-II maize
  • Zea mays
  • Transformation