Abstract:The aim was to obtain the compatible mycorrhizal fungi of lithophytic Paphiopedilum hirsutissimum using in situ seed baiting technique. Two natural small seedlings were harvested after in situ seed germination. The effective symbiotic fungi of genus Epulorhiza were isolated from these developed seedlings. In order to verify the effect of the isolated fungal strains on seed germination, one of strains, Phs34, was used to the in vitro symbiotic seed germination of P. hirsutissimum on the sterile habitat substrate. After cultured for 6 weeks, seeds inoculated with Phs34 started to develop into protocorms and average germination rate was (58.35±3.41)%, whereas seeds without the fungus failed to germinate. These indicated that the fungal strain Phs34 was compatible mycorrhizal fungus of P. hirsutissimum, and it will be helpful in seedling production and orchid conservation of P. hirsutissimum.