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Related Experiment Videos

T-DNA recombination and replication in maize cells.

Xiaoxia Zhao1, Isabelle Coats, Ping Fu

  • 1Transformation Research, Pioneer Hi-Bred International Inc., Johnston, IA 50131, USA.

The Plant Journal : for Cell and Molecular Biology
|August 29, 2003
PubMed
Summary

Replication of T-DNA in maize cells, driven by the wheat dwarf virus (WDV) system, promotes T-DNA recombination. This process, linked to viral replication, generates diverse recombinant molecules and activates marker genes.

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Area of Science:

  • Plant molecular biology
  • Virology
  • Genetics

Background:

  • Agrobacterium tumefaciens T-DNA transfer and integration are fundamental to plant transformation.
  • Understanding T-DNA replication and recombination mechanisms is crucial for improving plant biotechnology.

Purpose of the Study:

  • To investigate T-DNA recombination and replication in maize cells using the wheat dwarf virus (WDV) replication system.
  • To elucidate the relationship between T-DNA replication and recombination events.

Main Methods:

  • Transformation of 'black mexican sweet' (BMS) maize cells with T-DNAs harboring the WDV replication system.
  • Analysis of T-DNA recombination using a promoterless marker gene (gusA).
  • Utilizing mutant viral initiator genes (rep) to assess replication-dependent recombination.

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  • Sequencing of T-DNA border junctions to characterize recombination events.

    • Main Results:

    • T-DNA replication in maize cells, mediated by the WDV system, led to T-DNA amplification and recombination.
    • Recombination marker gene activation was time-dependent and increased exponentially.
    • Defective viral replication initiator genes (rep) abolished recoverable recombinant T-DNA formation.
    • Replicating T-DNAs served as substrates for site-specific and homologous recombination, generating a heterogeneous population of circular T-DNAs.
    • T-DNA recombination was linked to and stimulated by replication, exhibiting characteristics of illegitimate recombination.

    Conclusions:

    • T-DNA replication is intrinsically linked to and stimulates T-DNA recombination in maize cells.
    • The WDV replication system facilitates T-DNA amplification and generates diverse recombinant T-DNA molecules.
    • The observed recombination patterns share similarities with T-DNA integration into the plant genome but differ in border integrity.