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Related Experiment Videos

A versatile system for detecting transposition in Arabidopsis

N V Fedoroff1, D L Smith

  • 1Carnegie Institution of Washington, Department of Embryology, Baltimore, MD 21210.

The Plant Journal : for Cell and Molecular Biology
|February 1, 1993
PubMed
Summary

Researchers developed a versatile system using the Activator (Ac) transposable element in Arabidopsis thaliana to track transposon movement. This system efficiently identifies plants with reinserted transposons, aiding genetic studies.

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Area of Science:

  • Plant molecular genetics
  • Transposable elements
  • Arabidopsis thaliana research

Background:

  • The Activator (Ac) transposable element is active in dicots like Arabidopsis thaliana.
  • Arabidopsis is a model organism due to its small genome and rapid generation time.
  • Efficient methods are needed to track transposon activity in plants.

Purpose of the Study:

  • To develop a versatile system for identifying transposon excision and reinsertion events in plants.
  • To facilitate the study of transposon insertions near regulatory elements like promoters and enhancers.
  • To enable stable maintenance of transposons and transposase in experimental setups.

Main Methods:

  • Utilized the maize Activator (Ac) transposable element and marker genes.

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  • Incorporated a beta-glucuronidase (GUS) reporter gene with modified promoter sequences.
  • Employed herbicide chlorsulfuron for selection of transposon excision events.
  • Used additional selectable markers to distinguish transposed elements from transposase source.

    • Main Results:

    • Demonstrated a reliable and efficient system for identifying plants with transposed elements.
    • Somatic activation of the GUS gene indicates active transposition.
    • Chlorsulfuron resistance identifies progeny with excised transposons.
    • The system allows for separate or combined maintenance of transposon and transposase.

    Conclusions:

    • The developed system is effective for tracking transposon dynamics in Arabidopsis.
    • This tool enhances molecular genetic approaches in plant physiology and development.
    • The system's versatility supports various experimental designs for studying transposon insertion.