Figure 1.
The structures of nodulisporic acids unequivocally detected during this study.
See [12] for a detailed description of structures of the nodulisporic acids and their stereochemistry.
Table 1.
Living strains examined and their sequences. GenBank accession codes for previous and new sequences used during this work.
Figure 2.
Phylogenetic tree of Hypoxylon pulicicidum and related species of the Xylariaceae inferred from Bayesian analysis of a combined gene genealogy of α-actin, β-tubulin and ITS-5.8S-ITS2 genes.
Biscogniauxia anceps was designated the outgroup. Clade probability values/maximum likelihood bootstrap values are indicated respectively at the branches. Values <50 are designated by -. Bar represents 10 changes.
Figure 3.
Phylogenetic tree of living strains of Hypoxylon pulicicidum and related environmental strains and sequences deposited in GenBank inferred from Bayesian analysis of filtered ITS sequences.
Hypoxylon fendleri was designated the outgroup. Clade probability values/maximum likelihood bootstrap values are indicated respectively at the branches. Values <50 are designated by -. Bar represents 10 changes. Hypoxylon pulicicidum ITS1 sequence groups are identified as group1 with long repeated motifs, group 2 with intermediate repeated motifs, and group 3 with short repeated motifs. ITS1 base pair lengths are listed at the far right.
Table 2.
Preliminary identification of nodulisporic acids in fermentation extracts of strains of Hypoxylon investiens and H. pulicicidum by LC-MS database matching.
Table 3.
Detection and distribution of nodulisporic acid analogs by liquid chromatography high-resolution mass spectrometry among fermentation extracts of selected endophytic and teleomorph-derived strains of Hypoxylon pulicicidum.
Figure 4.
A, C, F, G: CLL0727; B, D, E, H: Holotype MJF 07147. A-C. Stromata. d. Close up of stromatal surfaces of a purplish mature stroma (left) and a blackish overmature stroma with slightly papillate ostioles (right). E, F. Stromata in vertical section showing perithecia with green contents (rehydrated in e). G, H. KOH-extractable pigments. Scale bar A-C = 5 mm; D-F 1 mm.
Figure 5.
Cultures and microscopic features of Hypoxylon pulicicidum.
A. MUCL 49879, from left to right, 14-d-old cultures on oatmeal agar, malt-yeast extract agar, 2% malt agar. B. MUCL 49879, conidiophores from SNA. C. MUCL 49879, conidiophores from SNA. D. MUCL 49879, conidiogenous cells and conidia from SNA. E. MUCL 49879, Aerial view of conidiophore from SNA. F. Ascus mounted in 1% SDS (MJF 07147). G. Ascus apical apparatus stained in Melzer’s reagent. H. Ascospores mounted in water (CLL 0727). Scale bar: F. 20 µm; H. 10 µm.
Figure 6.
Cultures and microscopic features of Hypoxylon pulicicidum, MF5954.
A. from left to right, 14-d-old cultures on oatmeal agar, malt-yeast extract agar, 2% malt agar. B. Conidiophore from SNA. C. Conidiophore from SNA, note dark pigment granules. D. Conidiogenous cells and conidia from SNA, note dark pigment granules. E. Aerial view of conidiophore from SNA. F. Aerial view of conidiophore from SNA. G. Conidiogenous cells and conidia from SNA.
Figure 7.
HPLC-UV chromatograms of Hypoxlyon pulicicidum and H. investiens.
9a. HPLC-UV chromatograms of stromatal methanol extracts of the H. pulicicidum (MJF 07147, Holotype). 9b. HPLC-UV chromatograms of stromatal methanol extracts of specimen of H. investiens s. str. (material from Taiwan, see Fig. S4). 9c. UV spectra of BNT and unknown components from H. pulicicidum. The unknown compound B in H. pulicicidum has a MW of 506 DA. 9d. UV spectra of components from H. investiens including daldinone A and another unknown compound D.