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RNA Silencing
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Molecular characterization of NbPAF encoding the alpha6 subunit of the 20S proteasome in Nicotiana benthamianaPlant Mol. Biol., 15
R. Lu, I. Malcuit, P. Moffett, M. Ruiz, Jack Peart, A. Wu, J. Rathjen, A. Bendahmane, L. Day, D. Baulcombe (2003)
High throughput virus‐induced gene silencing implicates heat shock protein 90 in plant disease resistanceThe EMBO Journal, 22
S. Dinesh-Kumar, R. Anandalakshmi, R. Marathe, Michael Schiff, Yule Liu (2003)
Virus-induced gene silencing.Methods in molecular biology, 236
Moonil Kim, Joon-Woo Ahn, U. Jin, Doil Choi, K. Paek, H. Pai (2003)
Activation of the Programmed Cell Death Pathway by Inhibition of Proteasome Function in Plants*Journal of Biological Chemistry, 278
R. Lu, A. Martín-Hernández, Jack Peart, I. Malcuit, D. Baulcombe (2003)
Virus-induced gene silencing in plants.Methods, 30 4
M. Ruiz, O. Voinnet, D. Baulcombe (1998)
Initiation and Maintenance of Virus-Induced Gene SilencingPlant Cell, 10
Y. Liu, M. Schiff, R. Marathe, S.P. Dinesh‐Kumar (2002b)
Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N‐mediated resistance to tobacco mosaic virusEMBO J., 30
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R. Saedler, I. Baldwin (2003)
Virus-induced gene silencing of jasmonate-induced direct defences, nicotine and trypsin proteinase-inhibitors in Nicotiana attenuata.Journal of experimental botany, 55 395
S. Holzberg, P. Brosio, C. Gross, G.P. Pogue (2002)
Barley stripe mosaic virus‐induced gene silencing in a monocot plantMol. Cells, 30
C. Burger, S. Rondet, P. Benveniste, H. Schaller (2003)
Virus‐induced silencing of sterol biosynthetic genes: identification of a Nicotiana tabacum L. obtusifoliol‐14alpha‐demethylase (CYP51) by genetic manipulation of the sterol biosynthetic pathway in Nicotiana benthamiana LAnnu. Rev. Phytopathol., 54
D. Robertson (2004)
VIGS vectors for gene silencing: many targets, many tools.Annual review of plant biology, 55
Virus‐induced gene silencing (VIGS) is an extremely powerful tool for plant functional genomics. We used Tobacco rattle virus (TRV)‐derived VIGS vectors expressed from binary vectors within Agrobacterium to induce RNA silencing in plants. Leaf infiltration is the most common method of agroinoculation used for VIGS but this method has limitations as it is laborious for large‐scale screening and some plants are difficult to infiltrate. Here we have developed a novel and simple method of agroinoculation, called ‘agrodrench’, where soil adjacent to the plant root is drenched with an Agrobacterium suspension carrying the TRV‐derived VIGS vectors. By agrodrench we successfully silenced the expression of phytoene desaturase (PDS), a 20S proteasome subunit (PB7) or Mg‐protoporphyrin chelatase (Chl H) encoding genes in Nicotiana benthamiana and in economically important crops such as tomato, pepper, tobacco, potato, and Petunia, all belonging to the Solanaceae family. An important aspect of agrodrench is that it can be used for VIGS in very young seedlings, something not possible by the leaf infiltration method, which usually requires multiple fully expanded leaves for infiltration. We also demonstrated that VIGS functioned to silence target genes in plant roots. The agrodrench method of agroinoculation was more efficient than the leaf infiltration method for VIGS in roots. Agrodrench will facilitate rapid large‐scale functional analysis of cDNA libraries and can also be applied to plants that are not currently amenable to VIGS technology by conventional inoculation methods.
The Plant Journal – Wiley
Published: Oct 1, 2004
Keywords: ; ; ; ;
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