Table 1.
Origin of isolates used in this study*.
Figure 1.
Best maximum-likelihood tree for the Pseudogymnoascus spp. used in this study (lnL = −10622.006982).
A concatenated alignment of the TEF-1, MCM7, and partial ITS genes was used to generate a Maximum-likelihood tree in RAxML using strains identified in Minnis and Lindner [5]. Values show bootstrap support from 500 bootstrap replicates; closed circles indicate ≥95% support at each branch point, while open circles indicate >70% support. The taxa shown in red were used in 10X tests, while the taxa in blue were used in triplicate tests. Geomyces auratus and members of the Leuconeurospora were used as the outgroups. Accession numbers for each sequence can be found in Tables 1 and S1.
Figure 2.
Temperature-dependent growth of environmental isolates of Pseudogymnoascus.
The plot shows the average growth rate of each taxon over 6 weeks. Error bars indicate the significant difference of each assay (n = 10). P. pan = P. pannorum.
Figure 3.
Growth of Pseudogymnoascus spp. on fulvic and humic acid extracts.
Boxplot boundaries show the first and third quartiles, with the mean as the centerline and whiskers as 1.5 times the inter-quartile distance. Outliers are plotted as points. P. des = P. destructans; P. pan = P. pannorum; Pen. p = Penicillium pinophilum; and Om = Oidiodendron maius.
Figure 4.
Relative enzyme activity for saprotrophic enzymes in P. destructans and other fungi.
Boxplot boundaries show the first and third quartiles, with the mean as the centerline and whiskers as 1.5 times the inter-quartile distance. Outliers are plotted as points. P. des = P. destructans; P. pan = P. pannorum; Pen. p = Penicillium pinophilum; and Om = Oidiodendron maius.
Figure 5.
Liquid assays of endoglucanase, β-glucosidase, and cellobiohydrolase.
Reducing sugars released from CMC (endoglucanase activity) and Avicel (β-glucosidase activity) when treated with fungal filter paper culture supernatant and p-nitrophenol release from pNPβG (cellobiohydrolase activity). Tests were performed in triplicate, and the results were plotted individually. The horizontal line indicates results for negative controls using culture filtrates and H2O.
Figure 6.
Relative enzyme activity of enzymes that may have pathogenic and saprotrophic function in environmental Pseudogymnoascus spp. and P. destructans.
A) 10x replicates testing lipase and urease activity at hibernaculum and ambient temperature. B) 3x replicates testing lipase and urease activity at ambient temperature. C) 10x replicates testing α-hemolysis activity on on tryptic soy agar/5% sheep’s blood after 8 weeks (environmental isolates) and 11 weeks (P. destructans). Boxplot boundaries show the first and third quartiles, with the mean as the centerline and whiskers as 1.5 times the inter-quartile distance. Outliers are plotted as points.