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Potato virus X as a vector for gene expression in plants
S Chapman1, T Kavanagh, D Baulcombe
1Sainsbury Laboratory, Norwich Research Park, UK.
The Plant Journal : for Cell and Molecular Biology
|July 1, 1992
Summary
Potato virus X (PVX) shows potential as a gene expression vector in plants. While gene replacement failed, inserting GUS into the genome enabled systemic spread and high-level gene expression.
Area of Science:
- Plant virology
- Molecular biology
- Biotechnology
Background:
- Potato virus X (PVX) is a plant virus with potential for genetic manipulation.
- Developing efficient gene vectors is crucial for plant biotechnology and research.
Purpose of the Study:
- To evaluate the suitability of PVX as a gene vector in plants.
- To test two distinct PVX-based constructs for gene expression and systemic spread.
Main Methods:
- Constructed two PVX variants: one with the GUS gene replacing the coat protein, and another with GUS inserted alongside a duplicated coat protein promoter.
- Inoculated protoplasts and plants (Nicotiana clevelandii) with viral constructs.
- Analyzed viral accumulation, systemic spread, and foreign gene expression (GUS).
Main Results:
- PVX with coat protein gene replacement showed poor accumulation and no systemic spread.
- PVX with GUS insertion and duplicated promoter exhibited reduced protoplast accumulation but achieved systemic infection.
- High-level GUS expression was observed in infected tissues, with significant retention of the foreign gene insert.
Conclusions:
- PVX coat protein is essential for efficient viral infection cycle stages.
- Gene replacement is unsuitable for PVX-based vectors.
- PVX is a promising vector for unregulated gene expression in plants, particularly when the foreign gene is integrated into the genome with appropriate regulatory elements.