Macrofungi were collected from Phukhieo-Nam Nao Forest Complex where Phukhieo Wildlife Sanctuary (PKWS), Phuluang Wildlife Sanctuary (PLWS), Nam Nao National Park (NNP), Phu Kradueng National Park (PKNP) and Phu Ruea National Park (PRNP) were selected. Species observation and specimen collection were conducted in the rainy season, 2 times a year, from 2013 to 2016. The results showed that specimens were identified into 54 families, 143 genera and 359 species, and highest species number was found in NNP. A large number of species belong to genera were found in Amanita, Russula and Marasmius respectively. These were further classified into five groups based on their roles, edible mushrooms (99 species), poisonous mushrooms (11 species) mycorrhizal mushrooms (122 species) and decaying mushroom (216 species). The new species with endemic status was found in PKWS, star-shaped fungi (Astraeus sirindhorniae). This study can provide a guideline manual for the study of biodiversity of mushrooms in the Phukhieo-Nam Nao Forest Complex. All collected specimens have been preserved in the mushroom herbarium of the Department of National Parks, Wildlife and Plant Conservation in order to take advantage of bio-organic compounds in the future.