Rhizoctonia solani causes damping-off, and root and crown rot of sugar beet (Beta vulgaris L.) and overwinters as sclerotia and mycelia. Research was conducted to determine how best to produce large quantities of sclerotia and mycelia in vitro, and compare their pathogenicity with traditionally used colonized barley grains to sugar beet in vitro and in vivo. The greatest number of sclerotia was produced on amended clarified V8 medium and sclerotia caused more disease compared to barley inoculum in the greenhouse. The bio-control potential of Penicillium pinophilum on R. solani AG2-2 on sugar beet was evaluated in vitro and in vivo. Results showed that the presence of P.pinophilum with R.solani reduced damping-off by 75% and thus have the potential to be developed as a bio-control agent for this pathogen.