Abstract
Virus-Induced Gene Silencing (VIGS) with Tobacco rattle virus (TRV) vector, which is frequently used as a functional analysis tool for plant genes, especially in Solanaceous plants, is also able to be used for the suppression of endogenous gene expression in Arabidopsis. However the reported method is not suitable for uniform treatment of multiple samples of Arabidopsis plants. In this study, we tried to establish the simple and efficient method of large-scale phenotype-dependent screening by VIGS with TRV vector (TRV-VIGS) harboring normalized Arabidopsis cDNA library. Silencing phenotype was observed in all Arabidopsis ecotypes we examined by TRV-VIGS. The establishment of efficient experimental procedure that induces gene silencing in multiple plant samples using vacuum infiltration was succeeded. TRV-VIGS of CAD1, which is known as a negative regulator of cell death, induced similar cell death phenotype as in cad1 mutant. We suggested that this established TRV-VIGS screening method is applicable to isolate cell death-related genes efficiently. Using this screening method, we have isolated several novel Arabidopsis genes involved in cell death.
