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Virus-induced gene silencing in tomato
Yule Liu1, Michael Schiff, S P Dinesh-Kumar
1Department of Molecular, Cellular and Developmental Biology, OML 451, Yale University, PO Box 208104, New Haven, CT 06520-8104, USA.
The Plant Journal : for Cell and Molecular Biology
|September 11, 2002
Summary
Tobacco rattle virus (TRV) enables efficient gene silencing in tomato plants for functional studies. A modified TRV vector facilitates easy cloning of tomato expressed sequence tags (ESTs) for large-scale analysis.
Area of Science:
- Plant Molecular Biology
- Virology
- Genetics
Background:
- Virus-induced gene silencing (VIGS) is a powerful tool for gene function studies.
- Tobacco rattle virus (TRV)-based VIGS has been established in Nicotiana benthamiana.
Purpose of the Study:
- To demonstrate the efficacy of TRV-based VIGS in tomato (Solanum lycopersicum).
- To develop a modified TRV vector for simplified gene cloning and large-scale functional analysis of tomato expressed sequence tags (ESTs).
Main Methods:
- Infection of tomato plants with recombinant TRV.
- Gene silencing of PDS, CTR1, CTR2, RbcS, and cLED3L14 using TRV VIGS.
- Construction of a modified TRV vector utilizing the GATEWAY recombination system.
- Cloning of tomato ESTs into the modified TRV vector.
Main Results:
- Recombinant TRV efficiently silenced target genes in tomato.
- Suppression of CTR1 resulted in a constitutive ethylene response and upregulation of CHITINASE B, mimicking Arabidopsis ctr1 mutants.
- The modified TRV vector enabled restriction- and ligation-free cloning of tomato ESTs.
Conclusions:
- TRV-based VIGS is effective for gene function studies in tomato.
- The modified GATEWAY-compatible TRV vector streamlines the process of cloning tomato ESTs.
- This system facilitates high-throughput functional genomics in tomato.