Fungal pure cultures identified with both classical morphological methods and through barcoding sequences are a basic requirement for reliable reference sequences in public databases. Improved techniques for an accelerated DNA barcode reference library construction will result in considerably improved sequence databases covering a wider taxonomic range. Fast, cheap, and reliable methods for obtaining DNA sequences from fungal isolates are, therefore, a valuable tool for the scientific community. Direct colony PCR was already successfully established for yeasts, but has not been evaluated for a wide range of anamorphic soil fungi up to now, and a direct amplification protocol for hyphomycetes without tissue pre-treatment has not been published so far. Here, we present a colony PCR technique directly from fungal hyphae without previous DNA extraction or other prior manipulation. Seven hundred eighty-eight fungal strains from 48 genera were tested with a success rate of 86%. PCR success varied considerably: DNA of fungi belonging to the genera Cladosporium, Geomyces, Fusarium, and Mortierella could be amplified with high success. DNA of soil-borne yeasts was always successfully amplified. Absidia, Mucor, Trichoderma, and Penicillium isolates had noticeably lower PCR success.
Myrmecridium hiemale sp. nov. was isolated from snow-covered alpine bare soil and is described as the first eurypsychrophilic species of this genus of filamentous fungi. Colony growth temperature experiments were carried out in the range 4-37 C. Morphological characteristics and colony appearance were in accordance with characteristics typical for Myrmecridium, but M. hiemale does not grow at temperatures of 25 C and above. Sequence analyses of the internal transcribed spacer and LSU rRNA D1/D2 regions indicated that the strain in question represents a distinct taxon within the genus Myrmecridium (Myrmecridiaceae, Sordariomycetes, Ascomycota). The type strain of M. hiemale is CBS 141017. A morphological description is provided, and a key is presented for the currently known taxa of Myrmecridium, a group of interesting fungi that are either saprobes or plant endophytes.
Here we describe two new species of the genus Penicillium section Torulomyces with solitary phialides. Penicillium poederi sp. nov. was isolated from volcanic soils in Iceland. Penicillium tirolense sp. nov. was isolated from a sporocarp of Serpula lacrymans. Both species are characterised by slow growth rates and the production of a brown soluble pigment on CYA, conidiophores with solitary ampulliform phialides with smooth-walled stipes and warty, globose conidia and with connectives without visible rings. The spores of. P. poederi are 2.5 µm diam, while the spores of P. tirolense are 2.0 µm diam. In a multigene phylogeny based on the ITS, BenA, CaM and RPB2 gene regions P. tubakianum and P. wollemiicola are the closest relatives of P. poederi. This species differs from P. tubakianum and P. wollemiicola by its growth rates and by its pigmentation. The holotype of P. poederi is IB2017/0007, while SF014017 (CBS 147622) is a culture derived from the holotype. The closest relatives of P. tirolense are P. austricola and P. riverlandense. It differs from P. austricola by lower growth rates on all tested media and temperatures and by its larger spores. It differs from P. riverlandense by lower growth rates and the absence of growth at 37 °C. The holotype of P. tirolense is IBF2019/0162, while SF015108 (CBS 147625) is a culture derived from the holotype.
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