ABSTRACT Two new Penicillium species are reported from soil of an agroforestry system and an Atlantic Rainforest fragment in Pernambuco, Northeast Brazil. Using a polyphasic approach, combining partial β-tubulin (BenA), calmodulin (CaM), internal transcribed spacer (ITS), and RNA polymerase II second largest subunit (RPB2) gene sequences and morphological characters, Penicillium gercinae sp. nov. (section Ramigena, series Georgiensia), and Penicillium stangiae sp. nov. (section Lanata-Divaricata, series Dalearum) are described. Descriptions based on morphological features are provided. Penicillium gercinae is phylogenetically closely related to P. georgiense; however, P. gercinae does not grow on Czapek yeast extract (CYA) and has monoverticillate and biverticillate conidiophores. Penicillium stangiae is phylogenetically closely related to P. penarojense and P. vanderhammenii. As diagnostic features, P. stangiae had slower growth on CYA (28-30 mm in 7 days) and on DG18 (6-7 mm in 7 days) and lacked the production of acid on creatine agar. This research contributes to the expansion of knowledge on the taxonomy and diversity of Penicillium species in Brazil.