Abstract
Both RNA and DNA viruses have been engineered to serve as vectors for transient silencing in intact plants. Host gene sequences carried by the virus are seen by the plant as “foreign,” and homologous gene-silencing machinery acts on both the viral vector RNA and the endogenous host gene mRNA. DNA viruses, such as geminiviruses, are advantageous for silencing because only their mRNAs are silenced and their DNA genomes continue to replicate and move. The conserved genome organization of geminiviruses and the fact that they can be cloned into Escherichia coli plasmids, propagated, and then inoculated into plants for infection simplifies the procedure for silencing specific chromosomal genes in intact plants. This chapter describes the development of a silencing vector from cabbage leaf curl virus for use in Arabidopsis and procedures for silencing two genes simultaneously.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408, 796–815.
Gutierrez, C. (2000) DNA replication and cell cycle in plants: learning from geminiviruses. EMBO J. 19, 792–799.
Hanley-Bowdoin, L., Settlage, S. B., Orozco, B. M., Nagar, S., and Robertson, D. (1999) Geminiviruses: models for plant DNA replication, transcription and cell cycle regulation. Crit. Rev. Plant Sci. 18, 71–106.
Hill, J. E., Strandberg, J. O., Hiebert, E., and Lazarowitz, S. G. (1998) Asymmetric infectivity of pseudorecombinants of cabbage leaf curl virus and squash leaf curl virus: implications for bipartite geminivirus evolution and movement. Virology 250, 283–292.
Ratcliff, F., Martin-Hernandez, A. M., and Baulcombe, D. C. (2001) Technical advance: tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J. 25, 237–245.
Gossele, V., Fache, I., Meulewaeter, F., Cornelissen, M., and Metzlaff, M. (2002) SVISS—a novel transient gene silencing system for gene function discovery and validation in tobacco plants. Plant J. 32, 859–866.
Angell, S. M. and Baulcombe, D. C. (1997) Consistent gene silencing in transgenic plants expressing a replicating potato virus X RNA. EMBO. J. 16, 3675–3684.
Kumagai, M. H., Donson, J., della-Cioppa, G., Harvey, D., Hanley, K., and Grill, L. K. (1995) Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc. Natl. Acad. Sci. USA 92, 1679–1683.
Dinesh-Kumar, S. P., Anandalakshmi, R., Marathe, R., Schiff, M., and Liu, Y. (2003) Virus-induced gene silencing, in Plant Functional Genomics Methods and Protocols, vol. 236 (Grotewold, E., ed.), Humana, Totowa, NJ, p. 287–293.
Atkinson, R. G., Bieleski, G. R., Gleave, A. P., et al. (1998) Post-transcriptional silencing of chalcone synthase in petunia using ageminivirus-based episomal vector. Plant J. 15, 593–604.
Turnage, M. A., Muangsan, N., Peele, C. G., and Robertson, D. (2002) Geminivirus-based vectors for gene silencing in Arabidopsis. Plant J. 30, 107–114.
Weigel, D. and Glazebrook, J. (2002) Arabidopsis: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Boyes, D., Zayed, A., Ascenzi, R., McCaskill, A., Hoffman, N., Davis, K., and Gorlach, J. (2001) Growth stage-based phenotypic analysis of Arabidopsis: a model for high throughput functional genomics in plants. Plant Cell 13, 1499–1510.
Koncz, C., Mayerhofer, R., Koncz-Kalman, Z., Nawrath, C., Reiss, B., Redei, G. P., and Schell, J. (1990) Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana. EMBO J. 9, 1337–1346.
Dellaporta, S. L., Wood, J., and Hicks, J. B. (1993) A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1, 19–21.
Jose, J. and Usha, R. (2003) Bhendi yellow vein mosaic disease in India is caused by association of a DNA Beta satellite with a begomovirus. Virology 305, 310–317.
Sambrook, J. and Russell, D. W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Kjemtrup, S., Boyes, D. C., Christensen, C., McCaskill, A. J., Hylton, M., and Davis, K. (2003) Growth stage-based phenotypic profiling of plants, in Plant Functional Genomics Methods and Protocols, vol. 236 (Grotewold, E., ed.), Humana, Totowa, NJ, pp. 427–441.
Kjemtrup, S., Sampson, K., Peele, C., Nguyen, L. V., Conkling, M. A., Thompson, W. F., and Robertson, D. (1998) Gene silencing from plant DNA carried by a Geminivirus. Plant J. 14, 91–100.
Peele, C., Jordan, C. V., Muangsan, N., Turnage, M., Egelkrout, E., Eagle, P., Hanley-Bowdoin, L., and Robertson, D. (2001) Silencing of a meristematic gene using geminivirus-derived vectors. Plant J. 27, 357–366.
Elmer, S. and Rogers, S. G. (1990) Selection for wild type size derivatives of tomato golden mosaic virus during systemic infection. Nucleic Acids Res. 18, 2001–2006.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc.
About this protocol
Cite this protocol
Muangsan, N., Robertson, D. (2004). Geminivirus Vectors for Transient Gene Silencing in Plants. In: Gott, J.M. (eds) RNA Interference, Editing, and Modification. Methods in Molecular Biology, vol 265. Humana Press. https://doi.org/10.1385/1-59259-775-0:101
Download citation
DOI: https://doi.org/10.1385/1-59259-775-0:101
Publisher Name: Humana Press
Print ISBN: 978-1-58829-242-1
Online ISBN: 978-1-59259-775-8
eBook Packages: Springer Protocols