Initiation of Plant Virus Infections by Agroinfiltration of Infectious Viral cDNAs (icDNAs)

Agroinfiltration uses Agrobacterium to deliver T-DNA-based gene expression constructs into plants. This chapter focuses on the standard method, specifically from the perspective of plant virus research, and describes a protocol for the initiation of virus infections in plants via infiltration of Agrobacterium strains carrying infectious viral cDNAs (icDNAs). The method outlines the culture and preparation of Agrobacterium for infiltration, the infiltration procedure, optimization of the optical density of the Agrobacterium suspension, and sampling of infected plants post-agroinfiltration. The advantages of the agroinfiltration method compared to traditional mechanical inoculation using sap from infected plants are discussed. The protocol is applicable for different pathosystems, although case-specific optimization of infiltration parameters and sampling is recommended.