Capsicum frutescens is known as a highly pungent chili pepper species, but an accession of C. frutescens showed very low capsaicinoid content. This few-pungent trait is novel and unique to 'S3212' in C. frutescens. To investigate the inheritance of the few-pungent trait, segregation of capsaicinoid content in F 2 and backcross populations (BC 1 ) crossed with high pungent C. chinense 'S3010' were observed. The segregation ratio indicated that fewpungency was controlled by a single recessive gene designated as the cf gene. Isolocus test between cf locus and C locus controlling capsaicinoid production was carried out using cleaved amplified polymorphic sequence (CAPS) marker SCY-800 linked to the C locus. CAPS pattern and few-pungency were not related, so the cf locus is different from the C locus. The single cf gene controlling few-pungency found in this study is expected to be used to develop new cultivars stable in few-pungency.