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Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors

Plant Molecular Biology volume 66, pages 587–598 (2008)Cite this article

Abstract

The Cre/loxP site-specific recombination system has been applied in various plant species including maize (Zea mays) for marker gene removal, gene targeting, and functional genomics. A BIBAC vector system was adapted for maize transformation with a large fragment of genetic material including a herbicide resistance marker gene, a 30 kb yeast genomic fragment as a marker for fluorescence in situ hybridization (FISH), and a 35S-lox-cre recombination cassette. Seventy-five transgenic lines were generated from Agrobacterium-mediated transformation of a maize Hi II line with multiple B chromosomes. Eighty-four inserts have been localized among all 10 A chromosome pairs by FISH using the yeast DNA probe together with a karyotyping cocktail. No inserts were found on the B chromosomes; thus a bias against the B chromosomes by the Agrobacterium-mediated transformation was revealed. The expression of a cre gene was confirmed in 68 of the 75 transgenic lines by a reporter construct for cre/lox mediated recombination. The placement of the cre/lox site-specific recombination system in many locations in the maize genome will be valuable materials for gene targeting and chromosome engineering.

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Acknowledgments

We thank Drs. C.M. Hamilton for BIBAC vectors (pCH20 and BIBAC1.Y30) and helper plasmid pCH32, D. Ow for the cre/lox recombination constructs pED97 and pHK52, P. Quail for the pAHC25 construct, and E. Richards for the pATt4 Arabidopsis telomere construct. We thank the University of Missouri DNA Core for sequencing. JMV thanks M. González-García for help in the preparation of some of the figures. Maize transformation experiments were conducted in the Plant Transformation Core Facility at the University of Missouri-Columbia. JMV was supported partially by an EMBO long-term fellowship. The funding was provided by a grant from the University of Missouri-Monsanto proteomics grant program and from NSF grant DBI 0423898. The materials described are available for research purposes from the corresponding author.

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Author notes

  1. Juan M. Vega

    Present address: Departamento de Genética, Facultad de Biología, Universidad Complutense, 28040, Madrid, Spain

  2. Akio Kato

    Present address: Faculty of Agriculture, Kyoto Prefectural University, Sakyo-ku, Kyoto-shi, Kyoto-fu, 606-0823, Japan

Affiliations

  1. Division of Biological Sciences, University of Missouri, Tucker Hall, Columbia, MO, 65211, USA

    Juan M. Vega, Weichang Yu, Fangpu Han, Akio Kato, Eric M. Peters & James A. Birchler

  2. Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, Columbia, MO, 65211, USA

    Zhanyuan J. Zhang

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  1. Juan M. Vega
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Correspondence to James A. Birchler.

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Vega, J.M., Yu, W., Han, F. et al. Agrobacterium-mediated transformation of maize (Zea mays) with Cre-lox site specific recombination cassettes in BIBAC vectors. Plant Mol Biol 66, 587–598 (2008). https://doi.org/10.1007/s11103-007-9276-2

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  • DOI: https://doi.org/10.1007/s11103-007-9276-2

Keywords

  • Agrobacterium-mediated transformation
  • B chromosome
  • BIBAC vector
  • Cre/loxP
  • Maize