Helvella lacunosa, besides its mycorrhizal importance for plant forest species, has a high commercial value. The commercial production of H. lacunosa represents two main advantages: Economical benefits for the communities able to produce it; and conservation of biodiversity in the forests by preventing the species overexploitation and maintaining the mushroom-plants micorrhizal relationship. Carrying out this perspective requires, in principle, to isolate the species mycelium, and then to develop the mycorrhizing technique that allows the production of the mushroom. Isolating the mycelium by the first time, however, involves testing several culture media in order to identify the potential requirements for inducing the myceliums growth. In the present work we tested, in laboratory, five different agar-based culture media to isolate H. lacunosas mycelium, and to evaluate its growing rate, analysis of covariance showed that there is a simple linear relationship between incubation time and the diameter. Indicating that the cumulative growth rate of the mycelium, was different between the culture media (F4, 240 = 17,008, p <0.001). The APD medi u m had the highest growth rate followed by AEA. In both growing media, the diameter of the mycelium converges the fifth day. The lowest growth rate was presented in the media AEM and AEP.