Abstract: In order to protect the wild resources ofPhlebopusand promote its artificial cultivation process，the morphological and molecular identification of the fruit body samples collected from Southwest China were carried out. Then，the nitrogen source and its concentration，inorganic salt type added into parent medium were screened，and the effect of the combination of vitamin B1 and different nitrogen sources on the mycelial growth was explored. The results showed that the tested strains were allPhlebopusportentosus.The best nitrogen source was ammonium chloride，and the best nitrogen source concentration was 0.5 g/L. The effect of vitamin B1 on the growth of mycelium was significant，only when combined with ammonium nitrate，it significantly promoted the growth of hypha，but not when combined with the other three nitrogen sources. There was no significant difference in mycelium growth between the two inorganic salt combinations，but from the perspective of economic cost，MgCl2/KCl combination was recommended. The optimal compositions were as follows：glucose，20 g；NH4Cl，0.5 g；MgCl2，1 g；KCl，1 g；potato，200 g；agar，20 g；H2O，1 L.

Key words: Phlebopusportentosus；molecular identification；colony diameter；medium；screening  ,