Tobacco Rattle Virus-Induced Gene Silencing in Cotton

Virus-induced gene silencing (VIGS), as a tool for plant reverse genetics, has been widely used in cotton for target gene function analysis. Compared with genetically transformed plants, the target gene expression level is reduced in the newly emerged leaves and can carry out phenotype identification after a few weeks of Agrobacterium infiltration. In this chapter, we describe a detailed protocol for Agrobacterium-mediated TRV-VIGS system for cotton gene function studies, with focus on designing primers, constructing TRV-target gene vectors via homologous recombination method, preparing and infiltrating Agrobacterium with TRV-VIGS, and identifying target gene silencing.