Virus-induced gene silencing in Solanum species.
Virus-induced gene silencing (VIGS) has been used routinely in Nicotiana benthamiana to assess functions of candidate genes and as a way to discover new genes required for diverse pathways, especially disease resistance signalling. VIGS has recently been shown to work in Arabidopsis thaliana and in tomato. Here, we report that VIGS using the tobacco rattle virus (TRV) viral vector can be used in several Solanum species, although the choice of vector and experimental conditions vary depending on the species under study. We have successfully silenced the phytoene desaturase (PDS) gene in the diploid wild species Solanum bulbocastanum and S. okadae, in the cultivated tetraploid S. tuberosum and in the distant hexaploid relative S. nigrum (commonly known as deadly nightshade). To test whether the system could be utilised as a rapid way to assess gene function of candidate resistance (R) genes in potato and its wild relatives, we silenced R1 and Rx in S. tuberosum and RB in S. bulbocastanum. Silencing of R1, Rx and RB successfully attenuated R-gene-mediated disease resistance and resulted in susceptible phenotypes in detached leaf assays. Thus, the VIGS system is an effective method of rapidly assessing gene function in potato.
Brigneti
Gianinna
G
The Sainsbury Laboratory, John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK.
Martín-Hernández
Ana M
AM
Jin
Hailing
H
Chen
Judy
J
Baulcombe
David C
DC
Baker
Barbara
B
Jones
Jonathan D G
JD
D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D010088:Oxidoreductases / Q000235:genetics; D010942:Plant Viruses / Q000472:pathogenicity*; D032322:Solanum / Q000235:genetics / Q000821:virology*; D011198:Solanum tuberosum / Q000235:genetics / Q000821:virology; D014026:Nicotiana / Q000235:genetics / Q000821:virology
D010088:Oxidoreductases; C022398:phytoene dehydrogenase