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Related Experiment Video

Updated: Jun 23, 2026

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes
10:28

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Published on: February 14, 2020

Whiskers-mediated maize transformation.

Joseph F Petolino1, Nicole L Arnold

  • 1Discovery Research, Cell Biology, Dow AgroSciences LLC, Indianapolis, IN 46268, USA. JFPetolino@dow.com

Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
PubMed
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Whiskers-mediated transformation enables DNA delivery into maize cells, expanding gene transfer capabilities. This method efficiently produces fertile transgenic maize plants from embryogenic cultures within nine months.

Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Genetics

Background:

  • Agrobacterium-mediated transformation has limitations for certain plant species.
  • Direct DNA delivery methods are crucial for expanding gene transfer capabilities.
  • Whiskers treatment offers a method for delivering macromolecules, including DNA, into plant cells.

Purpose of the Study:

  • To investigate the efficacy of whiskers-mediated transformation for maize.
  • To establish a protocol for generating stably transgenic maize plants using this method.

Main Methods:

  • Utilizing embryogenic maize tissue cultures as a source of totipotent cells.
  • Treating cultures with silicon carbide whiskers and DNA to facilitate gene transfer.
  • Employing transient gene expression assays to confirm DNA uptake.

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Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes
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  • Implementing in vitro selection with selectable marker genes for stable transformation.
  • Regenerating fertile plants from selected transgenic tissue cultures.

    • Main Results:

    • Transient gene expression observed within hours of whiskers treatment, indicating DNA uptake.
    • Successful generation of stably transgenic maize tissue cultures.
    • Recovery of fertile transgenic maize plants from treated cultures.
    • A complete transformation-to-fertile-seed timeline of approximately nine months.

    Conclusions:

    • Whiskers treatment is a viable and efficient method for maize transformation.
    • This technique expands gene transfer possibilities to plant species beyond the typical Agrobacterium host range.
    • The method is cost-effective and utilizes relatively simple equipment.