To identify fungal proteins involved in the arbuscular mycorrhizal symbiosis, root-inducing transferred-DNA transformed roots of carrot (Daucus carota L.) were in vitro inoculated with Glomus intraradices. Proteins extracted from the extra-radical fungus were analysed by two-dimensional gel electrophoresis. A fungal reference map displaying 438 spots was set up. Four proteins, among the 14 selected for tandem mass spectrometry analysis, were identified including a NmrA-like protein, an oxido-reductase, a heat-shock protein and an ATP synthase β mitochondrial precursor. The possible fungal origin of a MYK15-like protein found in mycorrhizal roots was further discussed. This is the first report of arbuscular mycorrhizal fungal protein identifications by using a proteomic approach.