Mushroom surveys and collections were conducted in the western and eastern forest areas in Cambodia, and then fungal biodiversity was analyzed by identifying mushrooms. One thousand and three hundreds eighty three specimens were identified by morphological and genetical characteristics, and were classified into 238 species, 160 genera, 52 families, 15 orders, and 3 phylums. The collected mushrooms were immersed in 70% ethyl alcohol for DNA extraction, and the rest of them were dried in the portable mushroom dryer for 12 hrs. Among these mushrooms, the genera Mycena (8.7%), Ganoderma (5.6%), Microporus (5.3%), Marasmius (4.2%), Marasmiellus (3.0%), Phellinus (2.5%), Trametes (2.5%), Hygrocybe (1.9%) and Pycnoporus (1.5%) were dominant. In the western Cambodia, 1,061 specimens were collected from Koh Kong forests, while 263 specimens were collected from the eastern Cambodia, Seima and Mondulkiri forests. Elevations of surveyed sites were ranged from 0 to 750 m above sea level. The number of species observed in the elevation of 251-500 m was the highest as compared to the other ranges of elevation. Daldinia concentrica, Microporus vernicipes, Microporus xanthopus, Pycnoporus coccineus, Stereum hirsutum, and Stereum ostrea were commonly distributed in all ranges of elevation, while the distribution of Ceratomyxa fruticulosa, Panus fulvus, Schizophyllum, Trametes versicolor, and Tyromyces chioneus were limited under 500 m. One hundred and forty one species including Amauroderma sp., Bjerkandera adusta, Trichaptum abietinum, and Tyromyces chioneus were collected only in Cardamom, while 20 species including Auricularia auricula-judae, Coriolopsis sanguinaria, Rigidoporus microporus, and Xylaria polymorpha were collected only in Seima. Ganoderma sp., Mycena sp., Marasmius sp., Microporus xanthopus, Phellinus sp., and Russula sp. were dominant species in both the western and eastern Cambodia. Species diversity indices in the eastern and western survey sites were 1.83 and 1.77, respectively, while evenness indices were 0.92 and 0.90. The species similarity index between two survey sites was 0.42.