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A stable RNA virus-based vector for citrus trees.

Alexey S Folimonov1, Svetlana Y Folimonova, Moshe Bar-Joseph

  • 1Department of Plant Pathology, University of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.

Virology
|July 27, 2007
PubMed
Summary
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Developing stable virus-based vectors for woody plants is crucial. This study created a Citrus tristeza virus (CTV)-based vector for stable foreign gene expression in citrus trees, demonstrating long-term utility.

Area of Science:

  • Plant molecular biology
  • Genomics
  • Virology

Background:

  • Virus-based vectors are essential tools for gene expression and functional screening in plants.
  • Existing vectors are primarily for herbaceous plants, lacking stability for woody plant research.
  • Woody plants require stable vectors due to longer systemic infection and analysis times.

Purpose of the Study:

  • To develop a stable Citrus tristeza virus (CTV)-based vector for transient foreign gene expression in citrus.
  • To utilize green fluorescent protein (GFP) as a reporter for gene expression analysis.
  • To engineer and test CTV vector strategies for enhanced stability and functionality in woody plants.

Main Methods:

  • Engineered CTV vector constructs by modifying open reading frames (ORFs) and subgenomic (sg) mRNA elements.

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  • Tested strategies included GFP ORF substitution, fusion with coat protein (CP), and expression from sg mRNA using CTV or Beet yellows virus controller elements (CE).
  • Evaluated vector replication, encapsidation, GFP expression in protoplasts, and performance in citrus plants over multiple passages and years.

    • Main Results:

    • Several CTV-based vector strategies were examined for their ability to express foreign genes.
    • The 'add-a-gene' strategy yielded highly stable vectors, maintaining GFP expression for over 4 years in citrus trees.
    • Engineered vectors demonstrated successful replication, encapsidation, systemic movement, and long-term gene expression in citrus.

    Conclusions:

    • Stable virus-based vectors are achievable for woody plants like citrus.
    • The developed CTV-based vectors offer a valuable tool for plant molecular biology and functional genomics in citrus.
    • Long-term gene expression and stability were achieved, overcoming limitations of previous vector systems.