Hevea brasiliensis is a mycotrophic tree for which root colonization by arbuscular mycorrhizal fungi (AMF) under in vitro or pot culture conditions can take several weeks. The reason for this slow colonization is still unknown, but the exudation of antifungal compounds such as hevein by the roots may be one of the causes. Here, the root colonization of Medicago truncatula, a highly mycotrophic plant, was assessed after 12 days of growth in the extraradical mycelium network of the AMF Rhizophagus irregularis in close vicinity of H. brasiliensis plantlets or Urtica dioica seedlings (also known to synthesize antifungal compounds of the hevein family). We hypothesized that a negative impact on the root colonization of a M. truncatula seedling developing close to H. brasiliensis and U. dioica may give indirect proofs for the exudation of inhibitory molecules. The percentages of total root colonization of M. truncatula were 30.1 % lower in the presence of H. brasiliensis than in the control plants, and 29.1 % lower in presence of U. dioica. The abundance of arbuscules in the roots of M. truncatula was also lower in plants grown in presence of H. brasiliensis plantlets than in the control plants. Similarly, the succinate dehydrogenase and the phosphatase activities measured in the extraradical mycelium of R. irregularis were significantly lower in the presence of both plants, compared with the controls. No root colonization was observed in H. brasiliensis and U. dioica within the time-frame of the experiments. The low root colonization of M. truncatula when grown in the presence of rubber or stinging nettle suggested the exudation of diffusible molecules which could also explain the delayed root colonization of H. brasiliensis and the absence of colonization of U. dioica.