A survey of the fynbos biome of South Africa resulted in the isolation and characterization of 61 distinct Penicillium species. ITS barcodes place six of these in section Lanata-Divaricata. Based on morphology and multigene phylogenies, species were identified as P. oxalicum, P. skrjabinii, and four that were previously unknown. Multigene phylogenies resolve the new species into three consistent clades. Species from these clades were compared morphologically, and the new species are described here as P. annulatum, P. curticaule, P. malacosphaerulum, and P. ortum. Penicillium annulatum (CBS 135126T) characteristically produces colonies on Czapek yeast autolysate (CYA) and yeast extract sucrose (YES) agar that sporulate in annular patterns and grow faster on CYA at 37 °C than those of closely related species, except for the faster growing P. rolfsii. Penicillium curticaule (CBS 135128T) most closely resembles P. raperi, but grows more restrictedly on CYA at 37 °C and produces shorter phialides. Penicillium ortum (CBS135668T) grows faster on CYA at 37 °C than P. cremeogriseum, its closest relative. Penicillium malacosphaerulum (CBS 135120T) produces cleisthothecia that give colonies on most media a yellowish straw color, similar to those of P. reticulisporum. However, faster growth on CYA at 37 °C distinguishes the new species from P. reticulisporum.