Thirty-three different ectomycorrhizae isolated from rhizosphere of 12 tree species have been tested for in vitro isolation on MMN and MEA synthetic media. Out of these, twenty-seven mycorrhizae have successfully been cultured on MMN and MEA media. Six ectomycorrhizae did not show any response on MEA medium. Successful isolation of most of the ectomycorrhizal species on both media show their non specificity for nutrients. Six species, that show no response on any of the media,  due to the nonavailability of particular nutrients in the medium. These ectomycorrhizal isolates have been tested for their growth, culture and hyphal characteristics so as to select the most suitable species having rapid growth rate in synthetic media.