Efficient virus-induced gene silencing in Arabidopsis using a 'one-step' TYMV-derived vector.
Virus-induced gene silencing (VIGS) is an important tool for the analysis of gene function in plants. This technique exploits recombinant viral vectors harbouring fragments of plant genes in their genome to generate double-stranded RNAs that initiate homology-dependent silencing of the target gene. Several viral VIGS vectors have already been successfully used in reverse-genetics studies of a variety of processes occurring in plants. Here, we show that a viral vector derived from Turnip yellow mosaic virus (TYMV) has the ability to induce VIGS in Arabidopsis thaliana, accession Col-0, provided that it carries an inverted-repeat fragment of the target gene. Robust and reliable gene silencing was observed when plants were inoculated simply by abrasion with intact plasmid DNA harbouring a cDNA copy of the viral genome, thus precluding the need for in vitro transcription, biolistic or agroinoculation procedures. Our results indicate that a 76 bp fragment is sufficient to cause gene silencing in leaves, stems and flowers, and that the TYMV-derived vector also has the ability to target genes expressed in meristematic tissues. The VIGS vector described here may thus represent an ideal tool for improving high-throughput functional genomics in Arabidopsis.
Pflieger
Stéphanie
S
Laboratoire de Virologie Moléculaire, Institut Jacques Monod, UMR 7592 CNRS-Universités Paris 6-Paris 7, 2 place Jussieu, 75251 Paris Cedex 05, France.
Blanchet
Sandrine
S
Camborde
Laurent
L
Drugeon
Gabrièle
G
Rousseau
Alice
A
Noizet
Mildred
M
Planchais
Séverine
S
Jupin
Isabelle
I
D017360:Arabidopsis / Q000235:genetics* / Q000821:virology; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors; D016679:Genome, Viral; D016254:Mutagenesis, Insertional; D030821:Plants, Genetically Modified / Q000235:genetics; D010957:Plasmids; D018749:RNA, Plant / Q000235:genetics; D017806:Tymovirus / Q000235:genetics*
D018076:DNA, Complementary; D004279:DNA, Viral; D018749:RNA, Plant