A Penicillium species isolated during a 1960s study on the ecology of fungi infecting Pinus radiata timber, and subsequently held in an in-house collection in Rotorua, New Zealand, was found to differ morphologically and in growth rate from two closely related Penicillium species. Phylogenetic analysis of the rDNA internal transcribed spacer (ITS), β-tubulin, calmodulin and RNA polymerase II second largest subunit regions (RPB2) confirmed this to be a new species closely related to Penicillium ochrochloron in the Rolfsiorum series of the Lanata-Divaricata section and Aspergilloides sub-genus. Micromorphologically, the new species is characterised by predominantly monoverticilliate and occasional divaricate or biverticilliate conidiophores and smooth-walled subglobose to slightly ovoid conidia with absence of conidiogenesis at 25 °C. This new species is described here as Penicillium rotoruae sp. nov. which has potential applications in biofuel and biorefining industry.