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Agrobacterium-mediated transformation of seedling-derived maize callus

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

Efficient production of seedling-derived Type I callus was demonstrated for several corn genotypes including commercial inbred lines. Seeds were germinated on MS-based medium containing 10 mg l−1 picloram and 3 mg l−1 6-benzylaminopurine, which induced the development of axillary buds in the area of coleoptilar node. Nodal sections of 7–10-day old seedlings were isolated, split longitudinally, and placed on callus induction medium supplemented with 2.2 mg l−1 picloram and 0.5 mg l−1 2,4-dichlorophenoxyacetic acid. For lines L4 and L9 the frequency of embryogenic callus induction was 38–42% based on calli per split nodal section. Frequency of callus induction from split nodal sections of seeds germinated on media without growth regulators was 0–3%. Seedling-derived callus of five genotypes was used for Agrobacterium-mediated transformation. Two constructs containing the green fluorescence protein gene and genes for either neomycin phosphotransferase II or glyphosate selection were used in transformation experiments. Transformation frequency varied from 2 to 11% and about 60% of the T0 plants had 1–2 copies of transgenes.

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Notes

  1. Proprietery lines might be available to the public via materials transfer agreement.

Abbreviations

IE:

immature embryo

BAP:

Benzyladenine

NAA:

α-Naphtaleneacetic acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

Picl:

Picloram

As:

acetosyringone

NPTII:

Neomycin phosphotransferase II

GFP:

Green fluorescent protein

Gly:

Glyphosate

CP4:

glyphosate-resistant enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4

MES:

2-Morpholinoethanesulfonic acid

Carb:

Carbenicillin

Par:

Paromomycin

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Acknowledgements

We thank Jay Harrison for his help with statistical analysis. We also thank Chuck Armstrong for his critical input on the manuscript.

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Authors and Affiliations

  1. Monsanto Company, 700 Chesterfield Parkway North, St. Louis, MO, 63017, USA

    Vladimir Sidorov, Larry Gilbertson, Prince Addae & David Duncan

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  1. Vladimir Sidorov
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  2. Larry Gilbertson
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  3. Prince Addae
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  4. David Duncan
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Correspondence to Vladimir Sidorov.

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Communicated by W. Parrott

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Sidorov, V., Gilbertson, L., Addae, P. et al. Agrobacterium-mediated transformation of seedling-derived maize callus. Plant Cell Rep 25, 320–328 (2006). https://doi.org/10.1007/s00299-005-0058-5

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