This study was conducted to establish an efficient screening method for radish (Raphanus sativus) cultivars that are resistant to black spot, which is caused by Alternaria brassicicola. Seven A. brassicicola isolates were selected and investigated for their ability to produce spores and pathogenicity. Of these isolates, A. brassicicola KACC 40036 and 43923 produced abundant spores in V-8 juice agar medium and showed pathogenicity and strong virulence on radish seedlings. We examined the resistance of 61 commercial cultivars of radish to A. brassicicola KACC40036, and found that there are no highly resistant radish cultivars; however, some cultivars, such as 'Geumbong' and 'Searom', showed weak resistance to A. brassicicola. For further study, we selected four radish cultivars that showed different disease responses to A. brassicicola KACC40036. According to the growth stage of the radish seedlings, inoculum concentration, and incubation temperature of radish, development of black spot on four cultivars has been investigated. The results showed that younger seedlings were more sensitive to A. brassicicola than older seedlings, and the disease severity depended on the concentration of the spore suspension. The disease severity of plants incubated in humidity chamber at 25°C was greater than that of plants grown at 20°C or 30°C. Taken together, we suggest the following method for screening for radish plants that are resistant to A. brassicicola: 1) inoculate 16-dayold radish seedlings with an A. brassicicola spore suspension (2.0 × 10 5 spores•mL-1) using the spray method, 2) incubate the inoculated plants in a humidity chamber at 25°C for 24 h and then transfer the plants to a growth chamber at 25°C with 80% relative humidity under a 12 h light/ dark cycle, and 3) assess the disease severity of the plants two days after inoculation. breeding, disease resistance, inoculation, Raphanus sativus 본 연구는 농림축산식품부 해양수산부 농촌진흥청 산림청 Golden Seed 프로젝트 채소종자사업단 (213002-04-4-SBc10, 213002-04-4-SBZ10)의 지원에 의해 수행되었음.