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Related Experiment Videos

A barley activation tagging system.

Michael A Ayliffe1, Margaret Pallotta, Peter Langridge

  • 1CSIRO Plant Industry, Box 1600, Clunies Ross Drive, Canberra, ACT, 2601, Australia. michael.ayliffe@csiro.au

Plant Molecular Biology
|April 13, 2007
PubMed
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Researchers developed a new activation tagging system in barley using the maize Ac/Ds transposable element. This system effectively activates gene expression and creates mutants, aiding in understanding gene function in cereals.

Area of Science:

  • Genetics
  • Molecular Biology
  • Plant Science

Background:

  • Activation tagging is a genomics tool for discovering gene function through over-expression.
  • Existing systems are mainly used in dicots, with limited options for cereals like barley.
  • Understanding gene function in large cereal genomes is challenging due to gene redundancy.

Purpose of the Study:

  • To develop and characterize an activation tagging system in barley using the maize Ac/Ds transposable element.
  • To assess the efficiency of transposition and gene activation by the UbiDs element.
  • To demonstrate the system's utility in generating both over-expression and knockout mutants.

Main Methods:

  • Utilized a modified Ds element (UbiDs) with maize polyubiquitin promoters for activation tagging in barley.

Related Experiment Videos

  • Analyzed transposition frequencies and gene activation in multiple independent transformant families.
  • Identified and analyzed insertional knockout mutants, including an allele of the granule bound starch synthase I (waxy) gene.

    • Main Results:

    • The UbiDs element transposed efficiently in barley families, with insertion frequencies up to 52%.
    • Transposon-mediated expression strongly activated transcription of adjacent flanking sequences in all analyzed cell and tissue types.
    • Successfully generated insertional knockout mutants, including a waxy allele that reduced amylose accumulation.

    Conclusions:

    • The developed barley activation tagging system is effective for generating dominant over-expression and knockout mutants.
    • This system provides a valuable tool for functional genomics in barley and other cereal crops.
    • It offers a novel approach to overcome challenges posed by gene redundancy in understanding gene function in cereals.