Penicillium diversity in Canadian bat caves, including a new species, P. speluncae

Abstract

Penicillium species were commonly isolated during a fungal survey of bat hibernacula in New Brunswick and Quebec, Canada. Strains were isolated from arthropods, bats, rodents (i.e. the deer mouse Peromyscus maniculatus), their dung, and cave walls. Hundreds of fungal strains were recovered, of which Penicillium represented a major component of the community. Penicillium strains were grouped by colony characters on Blakeslee's malt extract agar. DNA sequencing of the secondary identification marker, beta-tubulin, was done for representative strains from each group. In some cases, ITS and calmodulin were sequenced to confirm identifications. In total, 13 species were identified, while eight strains consistently resolved into a unique clade with P. discolor, P. echinulatum and P. solitum as its closest relatives. Penicillium speluncae is described using macroand micromorphological characters, multigene phylogenies (including ITS, beta-tubulin, calmodulin and RNA polymerase II second largest subunit) and extrolite profiles. Major extrolites produced by the new species include cyclopenins, viridicatins, chaetoglobosins, and a microheterogenous series of cyclic and linear tetrapeptides.

Keywords: Genealogical Concordance Phylogenetic Species Recognition (GCPSR) concept; Pseudogymnoascus destructans (Pd); Thysanophora; new taxon; secondary metabolites; sect. Fasciculata.

Fig. 1.

ML tree based on ITS, BenA, CaM & RPB2 showing identities and diversity of Penicillium associated with bats or bat caves. Bootstrap values ≥ 80% are shown above branches while thickened branches indicate 100 % support. Sequences obtained from ex-type cultures are indicated by ^T. Sequences obtained from strains during this study are indicated by blue text, while the new species, P. speluncae, is in bold blue text. The tree was rooted to Talaromyces pinophilus.

Fig. 1.

ML tree based on ITS, BenA, CaM & RPB2 showing identities and diversity of Penicillium associated with bats or bat caves. Bootstrap values ≥ 80% are shown above branches while thickened branches indicate 100 % support. Sequences obtained from ex-type cultures are indicated by ^T. Sequences obtained from strains during this study are indicated by blue text, while the new species, P. speluncae, is in bold blue text. The tree was rooted to Talaromyces pinophilus.

Fig. 2.

ML trees of Penicillium section Fasciculata, based on concatenated, BenA, CaM and RPB2 alignments, showing the relationship of P. speluncae within the section. PP and BS values ≥ 0.95/80 are shown above thickened branches (* = 1.00/100; - = <0.95/80). Sequences obtained from ex-type cultures are indicated by ^T. Strains of the new species characterised based on morphology and extrolites is indicated by bold blue text. Trees were rooted to Penicillium robsamsonia.

Fig. 3.

Line drawing ofPenicillium speluncae. Scale bar = 10 µm.

Fig. 4.

Penicillium speluncae. A. Colonies, from left to right, top row: CYA, MEA, YES, OA; bottom row: reverse on CYA, MEA, YES, CREA. B–F. Conidiophores. G. Conidia. Scale bars = 10 μm.