Orchids keep the ascomycetes outside: a highly diverse group of ascomycetes colonizing the velamen of epiphytic orchids from a tropical mountain rainforest in Southern Ecuador

Orchid mycorrhizal detection, based only on fungal isolation from roots, is biased due to difficulties in isolating the respective fungi. Previous investigations have shown that mostly ascomycetes, housed in the sheltered compartment of the velamen covering the roots of epiphytic orchids, are isolated on agar plates. Roots of 83 individual epiphytic orchid of Stelis hallii, S. superbiens, S. concinna, Stelis sp. and Pleurothallis lilijae were sampled in the Reserva Biologica San Francisco, Southern Ecuador. The velamen was partly removed and root tissue macerated and transferred to Petri dishes containing corn meal agar, malt extract agar and Melin Norkrans modified medium. DNA was extracted from the cultures and the 5.8S-ITS region and partial nrLSU sequenced. Phylogenetic analysis revealed members of 12 orders of ascomycetes associated with the roots of these orchids, a much higher diversity than previously known. Twelve isolates were related to the Helotiales. Our results have uncovered some of the fun...