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Stable wheat transformation obtained without selectable markers.

Hugo R Permingeat1, María L Alvarez, Gerardo D L Cervigni

  • 1Centro de Estudios Fotosintéticos y Bioquímicos (CONICET-Fund.M. Lillo-Universidad Nacional de Rosario), Suipacha 531, 2000 Rosario, Argentina. permingeath@arnet.com.ar

Plant Molecular Biology
|July 15, 2003
PubMed
Summary

Transgenic wheat plants can be developed without using a selectable marker gene. This study demonstrates successful gene integration and transmission to progeny, even without selective pressure during transformation.

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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Genetics

Background:

  • Selectable marker genes are traditionally used in plant transformation to identify successfully modified cells.
  • The presence of selectable marker genes in the final transgenic product can raise regulatory concerns and is often undesirable.

Purpose of the Study:

  • To investigate the possibility of generating transgenic wheat plants lacking selectable marker genes.
  • To evaluate gene integration and inheritance in the absence of selective pressure.

Main Methods:

  • Wheat transformation using particle bombardment with co-transformation experiments.
  • Application of selective pressure (hygromycin) versus no selective pressure.
  • Southern blot analysis to confirm gene integration.

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  • Progeny analysis to assess gene transmission and segregation.

    • Main Results:

    • Transgenic wheat plants lacking the selectable marker gene (hpt) were obtained, with some integrating only the desired gene (uidA).
    • The uidA gene was stably integrated and transmitted to progeny following Mendelian rules.
    • Gene segregation and loss of the selectable marker were observed even when selective agents were used.
    • A significant number of transgenic plants (23 out of 1016 bombarded explants) were generated without any selective pressure.

    Conclusions:

    • It is feasible to produce marker gene-free transgenic wheat.
    • Eliminating selective pressure during transformation can lead to successful generation of transgenic plants with stable gene integration.
    • This approach offers a more efficient route for developing transgenic crops for commercialization.