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Related Experiment Videos

Maize-targeted mutagenesis: A knockout resource for maize.

Bruce P May1, Hong Liu, Erik Vollbrecht

  • 1Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.

Proceedings of the National Academy of Sciences of the United States of America
|September 5, 2003
PubMed
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Researchers developed an efficient system for site-selected transposon mutagenesis in maize, creating a large seed library. Over half of mutations are suppressed when Mutator (Mu) activity is lost, impacting functional genomics studies.

Area of Science:

  • Plant genetics
  • Molecular biology
  • Genomics

Background:

  • Transposon mutagenesis is a key tool for gene discovery in maize.
  • Robertson's Mutator (Mu) is a widely used transposon system.
  • Efficient generation and analysis of large mutant libraries are crucial for functional genomics.

Purpose of the Study:

  • To establish an efficient system for site-selected transposon mutagenesis in maize.
  • To create a comprehensive library of transposon-mutagenized seed for the maize community.
  • To investigate the role of Mutator (Mu) activity in mutation stability and phenotype expression.

Main Methods:

  • Generation of 43,776 F1 plants using Mu pollen parents and self-pollination.
  • Development of a maize-targeted mutagenesis service utilizing PCR for insertion selection.

Related Experiment Videos

  • Establishment of an interactive database for managing mutant information (pedigree, sequence, phenotype).
  • Inhibition of Mu activity in F1 plants and monitoring of Mu activity in F2 for phenotype analysis.

    • Main Results:

    • A seed mutant frequency of 10-4 to 10-5 per F1 plant was achieved.
    • Somatic insertions were reduced by inhibiting Mu activity in most F1 plants.
    • Seed phenotypes were shown to depend on Mu activity, with false positives occurring even in lines with apparent low activity.
    • More than half of all mutations were suppressed upon loss of Mu activity.

    Conclusions:

    • The developed system provides an efficient resource for site-selected transposon mutagenesis in maize.
    • Mu activity significantly influences mutation stability and phenotype expression, with implications for epigenetic models.
    • Understanding Mu activity is critical for accurate interpretation of mutation effects in functional genomics research.