Potato virus X-induced gene silencing in leaves and tubers of potato.
Virus induced gene silencing (VIGS) is increasingly used to generate transient loss-of-function assays and has potential as a powerful reverse-genetics tool in functional genomic programs as a more rapid alternative to stable transformation. A previously described potato virus X (PVX) VIGS vector has been shown to trigger silencing in the permissive host Nicotiana benthamiana. This paper demonstrates that a PVX-based VIGS vector is also effective in triggering a VIGS response in both diploid and cultivated tetraploid Solanum species. We show that systemic silencing of a phytoene desaturase gene is observed and maintained throughout the foliar tissues of potato plants and was also observed in tubers. Here we report that VIGS can be triggered and sustained on in vitro micropropagated tetraploid potato for several cycles and on in vitro generated microtubers. This approach will facilitate large-scale functional analysis of potato expressed sequence tags and provide a noninvasive reverse-genetic approach to study mechanisms involved in tuber and microtuber development.
Faivre-Rampant
Odile
O
Programmes of Cell-to-Cell Communication, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, United Kingdom.
Gilroy
Eleanor M
EM
Hrubikova
Katarina
K
Hein
Ingo
I
Millam
Steve
S
Loake
Gary J
GJ
Birch
Paul
P
Taylor
Mark
M
Lacomme
Christophe
C
D001483:Base Sequence; D046508:Culture Techniques; D004171:Diploidy; D020868:Gene Silencing / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics* / Q000254:growth & development / Q000821:virology; D035281:Plant Tubers / Q000235:genetics* / Q000254:growth & development / Q000821:virology; D011123:Polyploidy; D017863:Potexvirus / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D011198:Solanum tuberosum / Q000235:genetics* / Q000254:growth & development / Q000821:virology
D010088:Oxidoreductases; C022398:phytoene dehydrogenase