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An aquatic hyphomycete, Dwayaangam sp., was isolated from superficially sterilized black spruce (Picea mariana) needles submerged in aerated water in a small glass chamber (microcosm). The internal transcribed spacer (ITS) sequence of this fungus and of a commonly encountered foliar endophyte isolated from P. mariana showed a high degree of similarity. When sporulation was induced in the microcosm, both the aquatic hyphomycete and the endophyte isolate produced similar aquatic conidia after 30 days, which is longer than previously documented in similar studies. Without the use of molecular tools, the link between the aquatic and endophytic phases of the fungus would have gone unnoticed. This is the first time that a fungal endophyte of conifer needles has been shown to have an aquatic phase. Its presence both as a foliar endophyte and a sporulating aquatic fungus suggests an alternating life cycle between the two environments.

Fungal foliar endophytes (hereinafter endophytes) were isolated on malt agar from black spruce ( Picea mariana (Mill.) BSP) needles from the Quebec boreal forest during 2 successive years to study their distribution along a latitudinal transect. Twenty sites were sampled in 2002, 8 of which were sampled again in 2003, in an area located between 47°N and 54°N in the province of Quebec. The endophytes were named whenever possible, using homologies of ITS rDNA sequences from GenBank. In 2002, 232 morphotypes were classified in 40 operational taxonomic units (OTUs), and in 2003, 93 morphotypes were classified in 16 OTUs, for a total of 44 OTUs. Isolation frequencies of the most common endophyte species were 28% for Lophodermium piceae (Fuchel) Höhn., 17% for Darkera parca Whitney, Reid & Piroz, and 9% for Dwayaangam colodena Sokolski & Bérubé. Of the 44 OTUs identified, 18 were unique to P. mariana and found on one site only, possibly constituting rare endemic species. There was some evidence that needles colonized with rare endophyte species were more abundant in the western and southern regions than in the northern region. This supports our hypothesis that as the diversity of tree species surrounding the P. mariana decrease from the southern region to the northern region, the diversity of endophytes also decreases.

Ribosomal nuclear genes are routinely utilized in the molecular identification of fungi. The variation in the multiple copies of these genes within each Glomeromycota strain and species reduces their usefulness for molecular characterization of arbuscular mycorrhizal fungi. To explore the potential of molecular tools for the identification of Glomus species, a multi-gene analysis approach was undertaken. Three protein-encoding genes were tested, namely elogation factor 1-α (765 bp), V-H+-ATPase VHA5 (1468 bp), and F0F1-ATPase β-subunit (621 bp). The latter is newly reported for the Glomeromycetes. Eleven species, including the type-specimen of Glomus irregulare Blaszk., Wubet, Renker & Buscot, a reference strain of G. intraradices N.C. Schenck & G.S. Sm. (DAOM 225240), and five strains of Glomus sp. formerly identified as G. intraradices, were analysed. These genes did not show polymorphisms within strains, and results indicated a close relationship between molecular identification and morphological characterization. Species with closely related spore morphological features, G. aggregatum N.C. Schenck & G.S. Sm., G. diaphanum Morton & Walker, G. irregulare , and Glomus sp. DAOM 197198, showed more than 99% nucleotide similarity, while the morphologically distinct species, G. cerebriforme McGee, G. clarum T.H. Nicolson & N.C. Schenck, G. claroideum N.C. Schenck & G.S. Sm., G. custos C. Cano & Dalpé, G. mosseae (T.H. Nicolson & Gerd.) Gerd. & Trappe, and G. proliferum Dalpé & Declerck, showed less than 97% similarity for at least one gene. A 100% molecular similarity for all three genes was found between G. irregulare and Glomus sp. DAOM 197198, confirming the new identity of the model arbuscular mycorrhizal fungus. Similarity thresholds for identification by DNA sequencing are discussed.