Tobacco bacterial wilt caused by Ralstonia solanacearum is one of the most destructive diseases suffered by tobacco in China. Streptomycin, calcium oxide, mancozeb, and synthetic fertilizer are four important non-target chemicals used frequently during the tobacco-growing period. This study evaluated the activities of four chemicals against R. solanacearum. Phenotypic pattern of R. solanacearum under pressure of those chemicals were also compared with control treatments. Bacterial growth of R. solanacearum was more sensitive to mancozeb, less sensitive to streptomycin, and not sensitive to calcium oxide and synthetic fertilizer. Metabolic reaction of R. solanacearum on Biolog GEN III Microplates was not or poorly affected by synthetic fertilizer or calcium oxide, while it was significantly inhibited by mancozeb and poorly inhibited by streptomycin. Substrates utilization of sucrose, α-D-glucose, D-fructose, D-galactose, D-sorbitol, myo-inositol, and p-Hydroxy-phenylacetic acid were inhibited by streptomycin and chemical sensitivities of pH 6, pH 5, tetrazolium violet, and tetrazolium blue were also changed by this chemical. No carbon source could be utilized by the pathogen under the pressure of mancozeb. Also, streptomycin had limited efficacy against R. solanacearum, while mancozeb could be used for bacterial wilt management. These new findings provide important information for tobacco bacterial wilt management in the field.