North American Fungi Volume 9, Number 9, Pages 1-13 Published October 14, 2014 A new species of Gomphus from southeastern United States Ronald H. Petersen1, Karen W. Hughes1, Jay Justice2 and David P. Lewis3 1 Ecology and Evolutionary Biology, University of Tennessee, Knoxville, TN 37996-1100. 2 16055 Michelle Dr., Alexander, AR 72002. 3262 County Road 3062, Newton, TX 75966 Petersen, R. H., K. W. Hughes, J. Justice, and D. P. Lewis. 2014. A new species of Gomphus from southeastern United States. North American Fungi 9(9): 1-13. http://dx.doi.org/10.2509/naf2014.009.009 Corresponding author: Ronald H. Petersen [email protected]. Accepted for publication October 1, 2014. http://pnwfungi.org Copyright © 2014 Pacific Northwest Fungi Project. All rights reserved. Abstract: Gomphus ludovicianus is proposed as a new species. It is described morphologically, placed phylogenetically and compared with G. crassipes from the Atlas Mountains of North Africa. Key words: Gomphales, cantharelloid fungi, taxonomy, phylogeography 2 Petersen et al. A new species of Gomphus. North American Fungi 9(9): 1-13 Introduction: Gomphus sensu lato has been transitions to transversions, the proportion of shown to be polyphyletic (Humpert et al.. 2001; invariant sites and the gamma distribution Hosaka et al, 2006; Giachini et al., 2010), with parameter estimated. One-hundred bootstrap one result being restriction of Gomphus sensu replicates were performed. Available ITS stricto to very few taxa (Giachini et al. 2010). One sequences for Gomphus and related Ramaria name, G. clavatus, is in use for basidiomata collections generated by this and other studies collected around the temperate Northern were too divergent to align easily. LSU sequences Hemisphere (Petersen, 1971). Giachini et al. also were unusually divergent from each other (2011) recognized only two additional names, G. but were alignable, although small differences in brunneus and G. crassipes, both from Africa. alignments could change observed tree structure With such a limited species list, it was surprising considerably. This is reflected by low bootstrap to find an undescribed species from southern scores at deeper nodes in the phylogeny. United States, not usually considered sub-boreal. Available ITS sequences and LSU sequences used The literature, however, described and illustrated to generate the LSU phylogeny are deposited in G. crassipes, which must be distinguished if the GenBank (Table 1). New World collections are to be proposed as new. Reciprocal monophyly of Gomphus clavatus and Materials and Methods: Morphological Gomphus ludovicianus was tested using procedures were described in Petersen and Rosenberg’s PAB statistic (Rosenberg 2007; Hughes (2010). Tissue response to 3% Potassium Masters et al. 2011) as implemented in Geneious hydroxide (KOH) was examined and color using the Species Delimitation plugin (Masters et changes noted. Microscopic characters were al., 2011). This statistic is the probability that a examined with light microscopy as well as phase putative species containing ‘A’ collections is contrast microscopy (PhC). monophyletic with respect to a putative sister clade containing ‘B’ collections. The ratio of the Color names enclosed in quotation marks are within-species genetic differentiation to from Ridgway (1912); those cited interspecies-genetic differentiation (Intra/Inter alphanumerically are from Konerup and ratio) was also examined within Geneious using Wanscher (1967). this plugin. Procedures for DNA extraction, PCR of the Results: ribosomal ITS and LSU regions and cloning of ITS PCR products was described in Hughes et al. Gomphus ludovicianus R.H. Petersen J. (Hughes et al. 2013). The data set used for Justice and D.P. Lewis Figs. 1-9. alignment was obtained by a blast search of GenBank accessions using Gomphus Mycobank no.: 808722 ludovicianus ITS and LSU sequences. This search recovered both Ramaria and Gomphus Holotype: Louisiana, Grant Parish, Kisatchie sequences, consistent with findings by Giachini National Forest, Catahoula Dist. vic. Intersection (2004) and Giachini and Castellano (2011) that LA 123 & FR 120 & CO 360, 31o 38.524’ N, 092o neither genus is monophyletic. PhyML 28.097’ W, 6.XII.2013, coll Jay Justice and Carl maximum likelihood (Guindon and Gascuel David, TFB 14476 (TENN 69161; holotype); 2008), for the ribosomal LSU dataset was performed in Geneious (Geneious 2005) using Macromorphology: Basidiomata (Figs. 1-4) the GTR model of evolution with the ratio of generally stout, obconical, usually depressed, Petersen et al. A new species of Gomphus. North American Fungi 9(9): 1-13 3 subpseudorhizal. Pileus 7-18 cm broad, shallowly basidiome flesh extremely friable, disarticulating convex when young, becoming applanate with into powder not supporting microscopic down-turned margin, innately coarsely scaly or examination. KOH applied to intact dried lumpy (like Hydnellum imbricatum, Turbinellus hymenium macroscopically unchanging (merely bonarii, see Figs. 2, younger basidiome, 4); pileus darkening to deep purple), immersion in KOH surface dull “benzo brown” (9D3) to “deep with cover slip quickly changing to deep orange- Quaker drab” (17B2) (compare left and right brown. Pileus tissue under subhymenium tightly surfaces in Fig. 1), bruising darker. Some surface interwoven, free (not adherent nor with over center paler. Paradermal scalp of dried gelatinized matrix); hyphae 3-6 µm diam, thin- to basidiome upper surface macroscopically dull firm-walled, conspicuously clamped. cream colored, instantly macro and microscopically red-orange in 3% aqueous KOH. Hymenium distinctly thickening, often in Fresh pileus and stipe flesh (Fig. 6) delicately semidefinable layers, appearing more congested mottled, “deep Quaker drab” to “ecru drab” and more pigmented in oldest layers, composed (16B2), naturally “yellow ocher” (4C7) when of paraphysoid basidioles, narrowly clavate bruised (by rotting or insect damage; Fig. 6). basidioles and basidia of various stages of Hymenophore delicately wrinkled (Figs. 3, 5), maturity. Basidioles (Fig. 8) 2.5-3.5 µm diam, outward on pileus with radial (longitudinal) linear, thin-walled, cylindrical, sometimes ridges higher and dominant, with subordinate apically lobed or branched, widening through anastomosing interveins, “purple gray” (23B3), development to elongate narrow-clavate shapes downward on stipe becoming gyrose without and developing scattered, small, refringent orientation (i.e. merismatoid), finally becoming guttules; mature basidia (Fig. 8) 59-62 × 9-12 m, smooth on upper stipe. Upper stipe surface dull clavate clamped, 4-sterigmate, effete after spore purplish gray mottled in small, ill-defined paler discharge and not totally collapsing or off-white areas, irregularly lumpy in contour, disintegrating and therefore forming succeeding matt. 3% KOH on fresh pileus surface = cherry hymenial layers. Basidiospores (Fig. 7B) (12-)14- red, similar on stipe, on hymenium only darker 17 × (4.5-)5-7 µm (Q = 1.87-2.82; Qm = 2.46; Lm = purple. Odor and taste negligible. 14.70 µm), ellipsoid with slightly depressed suprahilar region, minutely irregular in outline; Habitat and phenology: Forest of Quercus, contents with one or more amorphous, refringent Pinus palustris, Carya: known from central inclusions (not spherical, refringent oil droplets); Louisiana and southeastern Texas. ornamentation minutely rugulose, hardly definable, weakly cyanophilous. Micromorphology: Superficial pileus surface of young basidiomata detersile; hyphae of pileus Upper stipe surface a superficial dense thatch of scales or lumps 2-3.5 µm diam, tightly hyphae; hyphae very slender (2-2.5 µm diam), interwoven, with common hyphal termini tightly interwoven, gnarled, thin-walled, emergent and forming a delicate pruina; copiously branched with common acerose hyphal emergent termini cylindrical but irregular in tips protruding, interwoven portions apparently outline (Fig. 7A), often with suggestions of lobes involving abundant spherical bodies <1 µm diam, or branches. Tissue underlying pileus surface a obscuring accurate observation. Subtending layer of repent, tightly interwoven hyphae hyphal layer of strictly parallel, strictly without radial orientation; hyphae 2.0-4.5 µm longitudinal hyphae 3-3.5 µm diam, firm-walled, diam, inconspicuously clamped, thin-walled, occasionally but conspicuously clamped, perhaps unornamented; contents heterogeneous, adherent (disarticulating in bundles or sheets). obscuring observation of hyphal walls. Dried Stipe internal tissue generally longitudinally 4 Petersen et al. A new species of Gomphus. North American Fungi 9(9): 1-13 interwoven; hyphae 3-8 µm diam, firm- to thick- or merely absent in basidiomata in the collections walled (wall – 0.7 µm thick), conspicuously examined is unknown, but common spores clamped; ampulliform swellings common, -9 µm deposited on the pileipellis indicate that the diam, thick-walled (wall -1.5 µm thick, refringent; tissue forms, in fact, the true pileipellis surface. PhC), internally smooth to finely stalactitiformly ornamented. Especially DPL photos show pileus surface with the same coppery shades as commonly seen in Commentary: Although 3% aqueous KOH Gloeocantharellus purpurascens. KOH reactions applied to fresh or dried hymenium produces are identical, as they are with purple Ramaria only dark purple coloration, when a small portion taxa [R. fennica (P. Karst.) Ricken, R. of dried hymenium is immersed in KOH a color violaceibrunnea (C.D. Marr & Stuntz) R.H. change is immediate to