The large-scale functional analysis of genes in plants depends heavily on robust techniques for gene silencing. Virus-induced gene silencing (VIGS) is a transient gene silencing method for plants, triggered by the inoculation of a modified viral vector carrying a fragment of the gene targeted for silencing. Here we describe a VIGS protocol for rice, based on the Rice tungro bacilliform virus (RTBV, a DNA virus). We present an updated and detailed protocol for silencing of the gene encoding Phytoene desaturase in rice, using the RTBV-VIGS system.