First published online June 30, 2006; 10.1104/pp.106.084624
Plant Physiology 142:21-27 (2006)
© 2006 American Society of Plant Biologists
BREAKTHROUGH TECHNOLOGIES
Efficient Virus-Induced Gene Silencing in Arabidopsis1
Tessa M. Burch-Smith2,
Michael Schiff2,3,
Yule Liu2 and
S.P. Dinesh-Kumar*
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520–8103
Virus-induced gene silencing (VIGS) is a plant RNA-silencing technique that uses viral vectors carrying a fragment of a gene of interest to generate double-stranded RNA, which initiates the silencing of the target gene. Several viral vectors have been developed for VIGS and they have been successfully used in reverse genetics studies of a variety of processes occurring in plants. This approach has not been widely adopted for the model dicotyledonous species Arabidopsis (Arabidopsis thaliana), possibly because, until now, there has been no easy protocol for effective VIGS in this species. Here, we show that a widely used tobacco rattle virus-based VIGS vector can be used for silencing genes in Arabidopsis ecotype Columbia-0. The protocol involves agroinfiltration of VIGS vectors carrying fragments of genes of interest into seedlings at the two- to three-leaf stage and requires minimal modification of existing protocols for VIGS with tobacco rattle virus vectors in other species like Nicotiana benthamiana and tomato (Lycopersicon esculentum). The method described here gives efficient silencing in Arabidopsis ecotype Columbia-0. We show that VIGS can be used to silence genes involved in general metabolism and defense and it is also effective at knocking down expression of highly expressed transgenes. A marker system to monitor the progress and efficiency of VIGS is also described.
1 This work was supported by a National Science Foundation Plant Genome grant (grant no. DBI–0211872).
2 These authors contributed equally to the paper.
3 Present address: Ceres Inc., Thousand Oaks, CA 91320.
The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: S.P. Dinesh-Kumar (savithramma.dinesh-kumar{at}yale.edu).
www.plantphysiol.org/cgi/doi/10.1104/pp.106.084624
* Corresponding author; e-mail savithramma.dinesh-kumar{at}yale.edu; fax 203–432–6161.
Received June 2, 2006;
accepted June 20, 2006.
This article has been cited by other articles:
|
|
|
|
 
D. Orzaez, A. Medina, S. Torre, J. P. Fernandez-Moreno, J. L. Rambla, A. Fernandez-del-Carmen, E. Butelli, C. Martin, and A. Granell
A Visual Reporter System for Virus-Induced Gene Silencing in Tomato Fruit Based on Anthocyanin Accumulation
Plant Physiology,
July 1, 2009;
150(3):
1122 - 1134.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Renner, J. Bragg, H. E. Driscoll, J. Cho, A. O. Jackson, and C. D. Specht
Virus-Induced Gene Silencing in the Culinary Ginger (Zingiber officinale): An Effective Mechanism for Down-Regulating Gene Expression in Tropical Monocots
Mol Plant,
June 19, 2009;
(2009)
ssp033v1.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. C. Robbins, W. P. Heller, and M. R. Hanson
A comparative genomics approach identifies a PPR-DYW protein that is essential for C-to-U editing of the Arabidopsis chloroplast accD transcript
RNA,
June 1, 2009;
15(6):
1142 - 1153.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Dutta, S. Mohanty, and B. C. Tripathy
Role of Temperature Stress on Chloroplast Biogenesis and Protein Import in Pea
Plant Physiology,
June 1, 2009;
150(2):
1050 - 1061.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Padmanabhan and S. P. Dinesh-Kumar
Virus-Induced Gene Silencing as a Tool for Delivery of dsRNA into Plants
CSH Protocols,
February 1, 2009;
2009(2):
pdb.prot5139 - pdb.prot5139.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
L. C. Hileman and V. F. Irish
More is better: the uses of developmental genetic data to reconstruct perianth evolution
Am. J. Botany,
January 1, 2009;
96(1):
83 - 95.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. R. Scofield and R. S. Nelson
Resources for Virus-Induced Gene Silencing in the Grasses
Plant Physiology,
January 1, 2009;
149(1):
152 - 157.
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Garcion, A. Lohmann, E. Lamodiere, J. Catinot, A. Buchala, P. Doermann, and J.-P. Metraux
Characterization and Biological Function of the ISOCHORISMATE SYNTHASE2 Gene of Arabidopsis
Plant Physiology,
July 1, 2008;
147(3):
1279 - 1287.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Tzfira, S. V. Kozlovsky, and V. Citovsky
Advanced Expression Vector Systems: New Weapons for Plant Research and Biotechnology
Plant Physiology,
December 1, 2007;
145(4):
1087 - 1089.
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Spitzer, M. M. B. Zvi, M. Ovadis, E. Marhevka, O. Barkai, O. Edelbaum, I. Marton, T. Masci, M. Alon, S. Morin, et al.
Reverse Genetics of Floral Scent: Application of Tobacco Rattle Virus-Based Gene Silencing in Petunia
Plant Physiology,
December 1, 2007;
145(4):
1241 - 1250.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Dong, T. M. Burch-Smith, Y. Liu, P. Mamillapalli, and S. P. Dinesh-Kumar
A Ligation-Independent Cloning Tobacco Rattle Virus Vector for High-Throughput Virus-Induced Gene Silencing Identifies Roles for NbMADS4-1 and -2 in Floral Development
Plant Physiology,
December 1, 2007;
145(4):
1161 - 1170.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H.-C. Lu, H.-H. Chen, W.-C. Tsai, W.-H. Chen, H.-J. Su, D. C.-N. Chang, and H.-H. Yeh
Strategies for Functional Validation of Genes Involved in Reproductive Stages of Orchids
Plant Physiology,
February 1, 2007;
143(2):
558 - 569.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
