First published online July 15, 2005; 10.1104/pp.105.061861
Plant Physiology 138:2165-2173 (2005)
© 2005 American Society of Plant Biologists
PLANTS INTERACTING WITH OTHER ORGANISMS
Development of a Virus-Induced Gene-Silencing System for Hexaploid Wheat and Its Use in Functional Analysis of the Lr21-Mediated Leaf Rust Resistance Pathway1
Steven R. Scofield2,*,
Li Huang2,
Amanda S. Brandt and
Bikram S. Gill
United States Department of Agriculture, Agricultural Research Service, Crop Production and Pest Control Research Unit, West Lafayette, Indiana 47907 (S.R.S., A.S.B.); Department of Agronomy, Purdue University, West Lafayette, Indiana 47907 (S.R.S., A.S.B.); and Department of Plant Pathology, Throckmorton Plant Sciences Center, Kansas State University, Manhattan, Kansas 66506 (L.H., B.S.G.)
Virus-induced gene silencing (VIGS) is an important tool for the analysis of gene function in plants. In VIGS, viruses engineered to carry sequences derived from plant gene transcripts activate the host's sequence-specific RNA degradation system. This mechanism targets the RNAs of the viral genome for degradation, and as the virus contains transcribed plant sequence, homologous host mRNAs are also targeted for destruction. While routinely used in some dicots, no VIGS system was known for monocot plants until the recent report of silencing in barley (Hordeum vulgare) by barley stripe mosaic virus (BSMV). Here, we report development of protocols for use of BSMV to efficiently silence genes in hexaploid wheat (Triticum aestivum). The VIGS system was first optimized in studies silencing phytoene desaturase expression. Next, we used it to assay genes functioning in leaf rust resistance mediated by Lr21, which encodes a nucleotide binding site-leucine-rich repeat class resistance gene product. We demonstrated that infection with BSMV constructs carrying a 150-bp fragment of Lr21 caused conversion of incompatible interactions to compatible, whereas infection with a control construct or one that silences phytoene desaturase had no effect on resistance or susceptibility. Additionally, silencing the RAR1, SGT1, and HSP90 genes, known to be required in many but not all nucleotide binding site-leucine-rich repeat resistance pathways in diverse plant species, resulted in conversion to compatibility, indicating that these genes are essential in Lr21-mediated resistance. These studies indicate that BSMV-VIGS is a powerful tool for dissecting the genetic pathways of disease resistance in hexaploid wheat.
1 This work was supported by the U.S. Department of Agriculture, Agricultural Research Service Current Research Information System (project 3602–21220–008–00D; S.R.S. and A.S.B.), and by the Plant Biotechnology Center at Kansas State University and the Kansas Wheat Commission (funds to L.H. and B.S.G.).
2 These authors contributed equally to the paper.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.061861.
* Corresponding author; e-mail scofield{at}purdue.edu; fax 765–496–2926.
Received February 23, 2005;
returned for revision May 12, 2005;
accepted May 27, 2005.
This article has been cited by other articles:
|
|
|
|
 
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,
September 1, 2009;
2(5):
1084 - 1094.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. K. Bhullar, K. Street, M. Mackay, N. Yahiaoui, and B. Keller
Unlocking wheat genetic resources for the molecular identification of previously undescribed functional alleles at the Pm3 resistance locus
PNAS,
June 9, 2009;
106(23):
9519 - 9524.
[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]
|
|
|
|
|
|
|
Y. Meng, M. J. Moscou, and R. P. Wise
Blufensin1 Negatively Impacts Basal Defense in Response to Barley Powdery Mildew
Plant Physiology,
January 1, 2009;
149(1):
271 - 285.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Held, B. Penning, A. S. Brandt, S. A. Kessans, W. Yong, S. R. Scofield, and N. C. Carpita
Small-interfering RNAs from natural antisense transcripts derived from a cellulose synthase gene modulate cell wall biosynthesis in barley
PNAS,
December 23, 2008;
105(51):
20534 - 20539.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. C. Lee, I. S. Hwang, H. W. Choi, and B. K. Hwang
Involvement of the Pepper Antimicrobial Protein CaAMP1 Gene in Broad Spectrum Disease Resistance
Plant Physiology,
October 1, 2008;
148(2):
1004 - 1020.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Sindhu, S. Chintamanani, A. S. Brandt, M. Zanis, S. R. Scofield, and G. S. Johal
A guardian of grasses: Specific origin and conservation of a unique disease-resistance gene in the grass lineage
PNAS,
February 5, 2008;
105(5):
1762 - 1767.
[Abstract]
[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. Jing, D. Kornyukhin, K. Kanyuka, S. Orford, A. Zlatska, O. P. Mitrofanova, R. Koebner, and K. Hammond-Kosack
Identification of variation in adaptively important traits and genome-wide analysis of trait marker associations in Triticum monococcum
J. Exp. Bot.,
October 1, 2007;
58(13):
3749 - 3764.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. M. Crane and S. B. Gelvin
RNAi-mediated gene silencing reveals involvement of Arabidopsis chromatin-related genes in Agrobacterium-mediated root transformation
PNAS,
September 18, 2007;
104(38):
15156 - 15161.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Oikawa, A. Rahman, T. Yamashita, H. Taira, and S.-i. Kidou
Virus-induced gene silencing of P23k in barley leaf reveals morphological changes involved in secondary wall formation
J. Exp. Bot.,
July 1, 2007;
58(10):
2617 - 2625.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. K. Bhattarai, Q. Li, Y. Liu, S. P. Dinesh-Kumar, and I. Kaloshian
The Mi-1-Mediated Pest Resistance Requires Hsp90 and Sgt1
Plant Physiology,
May 1, 2007;
144(1):
312 - 323.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Cockram, H. Jones, F. J. Leigh, D. O'Sullivan, W. Powell, D. A. Laurie, and A. J. Greenland
Control of flowering time in temperate cereals: genes, domestication, and sustainable productivity
J. Exp. Bot.,
April 9, 2007;
(2007)
erm042v1.
[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]
|
|
|
|
|
|
|
E. A. Kellogg
Progress and challenges in studies of the evolution of development
J. Exp. Bot.,
October 1, 2006;
57(13):
3505 - 3516.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Travella, T. E. Klimm, and B. Keller
RNA Interference-Based Gene Silencing as an Efficient Tool for Functional Genomics in Hexaploid Bread Wheat
Plant Physiology,
September 1, 2006;
142(1):
6 - 20.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
