Phacidiaceae endophytes of Picea rubens in Eastern Canada

More than 100 fungal endophyte strains belonging to the family Phacidiaceae were isolated from surface-sterilized Picea rubens Sarg. needles collected from the Acadian Forest Region in New Brunswick, Canada. The strains were characterized morphologically by their asexual states, and phylogenetic analyses were conducted using the nuclear internal transcribed spacer rDNA (ITS) marker and the second largest subunit of ribosomal polymerase II (RPB2). Morphological and phylogenetic data revealed seven species: Darkera cf. parca; Strasseria geniculata; two novel Phacidium species: Phacidium dicosmoanum and Phacidium faciforme; and three novel monotypic genera described to accommodate distinct species: Calvophomopsis rubenticola, Cornibusella ungulata, and Gloeopycnis protuberans. Further analyses of Darkera spp. were performed with ITS and partial translation elongation factor 1-alpha (TEF1 alpha), and the results suggest that D. parca is a species complex. Phacidiaceae includes hundreds of known species that are unrepresented by sequence data; therefore, ITS sequences were generated from herbarium material including type specimens of Darkera parca, Phacidium lunatum, and specimens of Allantophomopsiella, Allantophomopsis, Bulgaria, Phacidium, and Pseudophacidium species. The description of novel species combined with morphological observations and reference sequences will facilitate the identification of conifer endophytes, both from specimens or cultures and in environmental sequence data, and improve our understanding of this large and mostly neglected family.