Pisolithus is a rare macro-fungal genus belonging to the family Sclerodermataceae and has been identified for the first time in Anbar. This puffball grew associated with Eucalyptus sp. tree and was collected during October 2013 at the campus of University of Anbar (UOA), Ramadi, which lies at 33.403457° N and 43.262189° E in dry conditions. This mushroom is considered to be ectomycorrhizal (ECM) and has an essential role in the physiology of Eucalyptus sp. This study added a new species to the biodiversity of macro-fungi in the arid and semi-arid area in Iraq.