Abstract

Materials and methods

Taxonomy

Gyromitra khanspurensis Jabeen & Khalid, sp. nov. – Figs. 1–2

Open in a separate window

Fig. 1

Gyromitra khanspurensis. Ascomata, holotype, LAH35074, bar 0.5 cm.

Open in a separate window

Fig. 2

Gyromitra khanspurensis, holotype, LAH35074. A. Ascospores. B. Asci. C. Paraphyses. Scale bars 10 μm.

MycoBank no.: MB 821408

Description. – Ascomata up to 3.4 cm high, hymenium free from the stipe; irregular, up to 2 cm high; 3.5 cm wide at the widest point; yellowish brown (5YR7/12) to brown (2.5YR5/10); surface smooth to wrinkled, becoming more wrinkled with age. – Stipe 3.5 × 1 cm; base slightly wider up to 1.3 cm, off-white, surface smooth. – Ascospores 14–17 × 7–8.5 μm, Q = 1.5–2, avQ = 1.8; smooth, ellipsoid, guttulate, apiculus short. – Asci up to 220 × 10 μm; 8-spored; cylindrical with a long narrow base; hyaline. – Paraphyses up to 140 × 6.5 μm clavate, narrowing towards base; hyaline. All tissues observed in 5 % KOH.

Etymology. – Named after Khanspur, the site of collection in Khyber Pakhtunkhwa province, Pakistan.

Holotypus. – PAKISTAN. Khyber Pakhtunkhwa province, Abbottabad district, Khanspur, near Cedrus deodara (Roxb. ex D. Don) G. Don (Pinaceae), 19 May 2014, leg. L. Choudry & A.N. Khalid, holotype SJ95 (LAH35074), ITS sequence GenBank MF116159.

Notes. – Gyromitra khanspurensis is characterized by its yellowish brown wrinkled hymenium free from the stipe, long clavate asci, and ellipsoid, hyaline ascospores. Morphologically it is closely related to G. gigas (Krombh.) Cooke, but differs by smaller ascospores than in G. gigas (26–40 × 11.5–15 μm) (Kuo 2012). Type material of G. gigas, which is the oldest name and has priority, is absent and there is some confusion regarding the spore size. Abbott & Currah (1997) synonymised the North American G. korfii (Raitv.) Harmaja and G. montana with G. gigas and consider the latter to have a broad range of ascospore size. The North American G. korfii is represented by a type specimen having an irregularly wrinkled hymenium borne on a ribbed stipe. Gyromitra khanspurensis bears more convoluted hymenium with a smooth stipe in comparison with G. korfii. Gyromitra khanspurensis also differs from G. korfii by the lack of an apiculus, which is prominent in G. korfii at mature stages (Methven et al. 2013). Morphologically, G. khanspurensis is also distinct from G. slonevskii V.P. Heluta. Gyromitra slonevskii bears a chestnut brown to liver coloured hymenium and white to brownish, rough and ribbed, slightly compressed stipe. Ascospore size is also comparatively larger (27–35 × 11–15 μm) in G. slonevskii (Hetula 2001). Molecular genetic sequence comparisons of the ITS also suggest G. khanspurensis as a distinct taxon (Fig. 3).

Open in a separate window

Fig. 3

Molecular genetic analysis of Gyromitra khanspurensis based on ITS sequences. The evolutionary history was inferred by the Maximum Likelihood method using Jukes-Cantor model in MEGA6. The analysis involved 36 nucleotide sequences. There were a total of 1337 positions in the final dataset. Sequence generated from LAH35074 marked with ●.

Authors: S. Jabeen & A.N. Khalid

Hapalocystis kickxii (Westend.) Voglmayr, comb. nov. – Fig. 4

Open in a separate window

Fig. 4

Hapalocystis kickxii (a–s). a, b. Stromata on dead corticated branch of Platanus orientalis, showing the translucent black perithecia; c–e. Transverse sections of pseudostromata, showing the perithecia with fusing perithecial necks; f. Vertical section of a pseudostroma showing the strongly depressed perithecia; g. Two ostiolar openings surrounded by discharged dark brown ascospores; h. Young asci with paraphyses; i. Ascus apex; j–n. Asci (m. two-spored, n. one-spored); o–s. Immature (o), almost mature (p, q) and fully mature (r, s) ascospores; Hapalocystis berkeleyi (t–a1). t–w. Asci (w. four-spored); x–a1. Ascospores. All vital in water. a–d, f, h, i, o-q WU 39964. e, g, j–n, r, s WU 39960 (epitype). t–a1 WU 39963. Scale bars: a, b, g 500 μm; c–e 400 μm; f 200 μm, h–a1 20 μm.

MycoBank no.: MB 823505

Basionym. – Hypoxylon kickxii Westend., Bull. Acad. R. Sci. Belg., Cl. Sci. 19(3): 112 (1852).

Synonyms. – Aglaospora kickxii (Westend.) Kuntze, Revis. gen. pl. (Leipzig) 3(2): 441 (1898).

Hapalocystis berkeleyi var. kickxii (Westend.) M.E. Barr, Mycol. Mem. 7: 193 (1978).

Prosthecium hapalocystis var. kickxii (Westend.) Wehm., Revision of Melanconis, Pseudovalsa, Prosthecium & Titania: 107 (1941).

Pseudovalsa kickxii (Westend.) Sacc., Syll. fung. (Abellini) 2: 139 (1883).

Valsa kickxii (Westend.) J. Kickx f., Fl. Crypt. Flandres (Paris) 1: 322 (1867).

Description. – Pseudostromata 1– 1.6 mm diam., immersed in the outermost layer of the bark of dead branches, flat or slightly convex, circular, separate, gregarious, sometimes confluent, perithecial outlines sometimes translucent through bark. Stromatic tissues sparse, consisting of loose light brown hyphae growing between bark and perithecia. – Perithecia 4–12 per stroma, black, pyriform, strongly depressed to lenticular, (250)300–600(750) μm diam., 130–170 μm high, monostichous, circinately arranged, with ostiolar necks of perithecia fused in a single central ostiole. – Ostiole (93)105–135(145) μm diam. (n=20), dark, not projecting, with hyaline, filamentous periphyses embedded in a gel matrix. – Paraphyses of broad bands 4–9 μm wide, septate, deliquescent at maturity. – Asci detached from the perithecial wall at maturity, (119)133–183(224) × (45)47–56(62) μm (n=28), broadly fusoid, with a 5–70 μm long stipe and an inconspicuous apical ring, containing 4 (rarely 1–2) ascospores. – Ascospores (42)48–61(91) × (14)17–23(33) μm, l/w = (2.3)2.5–2.9(3.2) (n=80), subhyaline to olive brown when fresh, dark brown with age, ellipsoid to slightly obovoid, asymmetric, rarely straight to usually distinctly curved, (3)4–5 septate, strongly constricted at the primary and strongly to not constricted at the secondary septa, with broadly rounded ends, with tubular gelatinous, slightly tapering appendages (18)23–31(37) × (5)6–7.5(9) μm long at each end; cells of unequal size, multiguttulate when vital; wall ca. 1–1.5 μm thick, finely verruculose, hyaline when young, becoming olive to dark brown at maturity, swelling, surrounded by a sharply delimited, irregularly swelling, up to 6 μm thick subhyaline to brownish gelatinous sheath.

Asexual morph not observed.

Habitat. – On dead corticated branches of Platanus spp. (P. ×hispanica, P. orientalis).

Distribution. – Widely distributed in Europe; known e.g. from Austria, Belgium, Germany, France, Greece, Netherlands, Spain, United Kingdom.

Material examined. – Hapalocystis kickxii: AUSTRIA. Wien, Landstraße, Botanical Garden of the University of Vienna (HBV), on corticated branches of Platanus orientalis and P. ×hispanica, 3 March 2008, leg. H. Voglmayr (WU 39960, epitype of Hypoxylon kickxii here designated; ex epitype culture D75 = CBS 123818; MBT379570) ibid., 10 November 2017, leg. H. Voglmayr (WU 39964); Währing, Türkenschanzpark, on corticated branches of Platanus ×hispanica, 11 November 2017, leg. H.Voglmayr (WU 39965). BELGIUM. Courtrai, Saint George park, on branches of Platanus orientalis, without date and collector, Westendorp & Wallays, Herb. Crypt. Belg. no. 714 (BR 5020097352610, lectotype of Hypoxylon kickxii here designated; MBT379569). FRANCE. Dept. Alpes-de-Haute-Provence (04), Gorge du Verdon near Rougon, on corticated twig of Platanus orientalis, 1 August 2008, leg. H. Voglmayr (WU 39961, culture D83 = CBS 124481); France, Dept. Côte d’Or (21), Mirebeau-sur-Bèze, Terrain de sports, on corticated branches of Platanus ×hispanica, 6 May 2015, leg. A. Gardiennet AG15025 (WU 39962, culture D108); GREECE. Crete, SE Rethymno, Kaloniktis, 7 June 2015, leg. H. Voglmayr & W. Jaklitsch (no specimen preserved). Hapalocystis berkeleyi: Austria, Wien, Währing, Türkenschanzpark, on corticated branches of Platanus ×hispanica, 11 November 2017, leg. H. Voglmayr (WU 39963).

Notes. – Within Hapalocystis, H. kickxii is well characterised by usually four-spored asci and five- to six-celled ascospores which become olive to dark brown at maturity; for a key to species and discussion on the genus Hapalocystis see Jaklitsch &Voglmayr (2004). Despite its distinctive features, it was reduced to a variety by Wehmeyer (1941, as Prosthecium berkeleyi var. kickxii), based on depauperate herbarium material which he considered to be abnormally developed. This was subsequently also followed by Barr (1978) who classified it as Hapalocystis berkeleyi var. kickxii. However, H. berkeleyi differs markedly by usually eight-spored asci and three-celled ascospores (Fig. 4t–a1). In addition, H. berkeleyi has distinctly smaller ascospores (28–34 × 11–16 μm vs. 42–68(91) × 14–25(33) μm in H. kickxii; measured without gel sheath; values in brackets in H. kickxii refer to a significantly larger ascospore from a monosporic ascus). Also the gelatinous appendages are of different size and shape (5–12 × 7–11 μm and blunt rectangular with broadly rounded ends in H. berkeleyi; 18–37 × 5–9 μm and long-tubular with tapering, narrowly rounded to subacute ends in H. kickxii). The presence, shape and size of ascospore appendages has been shown to be of high diagnostic value for species delimitation in Diaporthales (e.g.Voglmayr & Jaklitsch 2008, 2014; Voglmayr et al. 2012, 2017) but can often be only well-seen in fresh material. Phylogenetic analyses (Fig. 5) confirm that H. kickxii is clearly distinct from H. berkeleyi at species level. Besides the typical four-spored asci, few two- or one-spored asci (Fig. 4m, n) can occasionally be observed, with distinctly larger ascospores in the latter. Similarly, in H. berkeleyi four-spored asci with slightly larger ascospores are rarely present (Fig. 4w). However, apart from slightly to distinctly larger sizes, ascospores of these asci with reduced spore numbers are typical for the respective species.

Open in a separate window

Fig. 5

Phylogram of one of two best MP trees 471 steps long revealed by PAUP from an analysis of the ITS-LSU rDNA matrix of Sydowiellaceae, with Coryneum (Coryneaceae) and Asterosporium asterospermum (Asterosporiaceae) selected as outgroup, showing the status of Hapalocystis kickxii (bold) as distinct species, * denoting node collapsed in the strict consensus of both MP trees. MP bootstrap support above 50 % is given above the branches.

Ecologically, H. berkeleyi and H. kickxii share the same hosts and can co-occur on the same branches, but H. berkeleyi has been more commonly recorded from Platanus ×hispanica and H. kickxii from Platanus orientalis. Additional Hapalocystis species are found on Platanus in North America (Barr 1978, Jaklitsch & Voglmayr 2004), but except for H. occidentalis no sequence data are yet available.

The type collection was distributed as Westen-dorp & Wallays, Herb. Crypt. Belg. no. 714, and specimen BR 5020097352610 is here selected as lectotype. As the type collection is depauperate (Wehmeyer 1941; B. Declercq, unpubl. notes) and in order to stabilise the name, a recent well-developed specimen, for which a culture and sequence data are available, is here selected as lectotype.

Author: H. Voglmayr

Paramyrothecium guiyangense Y. Chen, Yong Wang bis & Y.L. Jiang, sp. nov. – Fig. 6

Open in a separate window

Fig. 6

Paramyrothecium guiyangense (HGUP 2016-8001, HGUP 2016-8002 and HGUP 2016-8007) a, b. Colonies were grown on PDA (a, from above; b, from below). c. Mycelium. d–f. Conidiophores and conidiogenous cells with conidia. g. Conidia. Scale bars 10 μm.

MycoBank no.: MB 819908

Description. – Sexual morph undetermined; Asexual morph: conidiomata sporodochial, superficial, cupulate, outline oval, 20–160 μm diam., with setose fringe surrounding a slimy mass of olivaceous green conidia. Setae arising from sporodochia, thin-walled, hyaline, 1−3-septate, smooth, straight to flexuous, becoming sinuous above the apical septum, 60–120 μm long, 1−3 μm wide, tapering to an acutely rounded apex. – Conidiophores hyaline, aseptate, smooth, 10−60 × 1−3 μm, growing from the basal stroma, stipe and branched in a whorl of 1−4 conidiogenous cells arising apically. – Conidiogenouscells hyaline, phialidic, subcylindrical, collarettes with conspicuous periclinal thickening, smooth, straight to slightly curved, 8–18 × 1.6−2.7 μm. – Conidia aseptate, hyaline, smooth, cylindrical to subcylindrical, 6.6−9.0 × 2−3 μm (n = 30, = 8 × 2.5 μm), rounded at both ends.

Colonies scattered on PDA with abundant white to pale luteous aerial mycelium. Sporodochia with slimy olivaceous green conidial masses formed on the surface of the medium. Cultures pale luteous in reverse.

Etymology. – guiyangense, referring to the collection locality.

Habitat and distribution. – Agricultural soil, China.

Holotypus. – CHINA. Guizhou Province, Guiyang City, Huaxi District, Yangguang park, from soil, 10 August 2016, leg. Y. Chen [holotype, GUCC 201608S01; sequences: KY196193 (cmdA), KY126418 (ITS), KY196209 (LSU), KY123426 (tef1), KY196201 (tub2)].

Further material examined. – CHINA. ibid. (HGUP 2016-8001, HGUP 2016-8002, HGUP 2016-8007, holotype, GUCC 201608S01); living culture, HGUP 2016-8002. Sequences accession numbers: HGUP 2016-8001 KY196192 (cmdA), KY126417 (ITS), KY196208 (LSU), KY126425 (tef1), KY196200 (tub2). HGUP 2016-8007 KY196198 (cmdA), KY126423 (ITS), KY196214 (LSU), KY126430 (tef1) and KY196206 (tub2).

Notes. – Paramyrothecium sp. (HGUP 2016-8001, HGUP 2016-8002, HGUP 2016-8007) clustered into a highly supported cluster (100%/100%/1.0) with P. foliicola, P. breviseta, P. roridum and P. verruridum. However, three strains could be clearly distinguished from these, forming an independent branch (Fig. 7). Morphologically, three strains had shorter conidiogenous cell (8–18 μm) and longer conidia (8 × 2.5 μm) among species in the branch. Based on their phylogeny and morphology, three of these strains were identified as belonging to the new species P. guiyangense.

Open in a separate window

Fig. 7

A Paramyrothecium consensus tree generated using sequence data of the cmdA, ITS, LSU, rpb2, tef1 and tub2 genes, with Alfaria ossiformis as an outgroup (CBS 324.54). Maximum likelihood, Maximum parsimony, and Bayesian approaches were used, but only the maximum likelihood tree is shown. Values of maximum likelihood and maximum parsimony bootstrap ≥ 70 % (MLBS/MPBS) with the third values of posterior probabilities (BI PP ≥ 0.95) are presented above or below the branches, respectively. Ex-type and ex-epitype strains are shown in bold, black text. Those strains used in this study and their support values are shown in red.

Paramyrothecium species are saprobe, weakly pathogenic fungi, able to grow in air, sands, soil and on plants, worldwide (Lombard et al. 2016). Lombard et al. (2016) performed a phylogenetic analysis of Myrothecium-like species using the cmdA, ITS, rpb2, and tub2 genes and identified twelve Myrothecium-like species (two Myrothecium species [M. acadiense (CBS 123.96), Seifert et al. (2003), M. roridum (CBS 357.89), Tulloch (1972)]; ten Myrothecium-like species [M. breviseta (CBS 544.75), M. cupuliforme (CBS 127789), M. foeniculicola (CBS 331.51), M. foliicola (CBS 113121), M. humicola (CBS 127295), M. nigrum (CBS 116537), M. parvum (CBS 257.35), M. tellicola (CBS 478.91), M. terrestris (CBS 564.86), and M. viridisporum (CBS 873.85)]) that formed a conspicuous independent clade with strong bootstrap values. Combined with the special long cylindrical-shaped conidia, Lombard et al. (2016) proposed the genus Paramyrothecium within the family Stachybotryaceae and Paramyrothecium roridum (Tode) L. Lombard & Crous was suggested as type species for Paramyrothecium, whose characteristics include: Sporodochia cupulate, surficial, covered by a slimy mass of olivaceous green to dark green conidia; setose, 1–4 septate, thin-walled, hyaline, straight or sinuous; conidiophores branched, smooth, hyaline, macronematous; conidiogenous cells phialidic, hyaline or darker at the apex, surface smooth or lightly guttulate, cylindrical or sub-cylindrical special collarettes; conidia cylindrical or ellipsoidal, straight or curve, smooth and tinting hyaline or pale green.

Paramyrothecium verruridum Y. Chen & Yong Wang bis, Y.L. Jiang, sp. nov. – Fig. 8

Open in a separate window

Fig. 8

Paramyrothecium verruridum (HGUP 2016-8006). a, b. Colonies grown on PDA (a, from above; b, from below). c. Mycelium and conidiomata. d–f. Conidiophores and conidiogenous cells with conidia. Scale bars = 10 μm. g, h. Conidia. Scale bars 5 μm.

MycoBank no.: MB 819909.

Description. – Sexual morph, undetermined; asexual morph, conidiomata sporodochial, superficial, cupulate, outline oval or irregular, 20−36 μm diam., white setose fringe surrounding a mass of slimy conidia, olivaceous green to dark green. Setae growing from sporodochia, thin-walled, hyaline, 1−3-septate, smooth, straight or flexuous becoming sinuous at the apical septum, 40–120 μm long, 2−3 μm wide, tapering to a sharply rounded apex. – Conidiophores hyaline, aseptate, smooth, 20−40 × 1.5−2.5 μm, growing from the basal stroma, one whorl of 2–8 conidiogenous cells growing apically. – Conidiogenous cells phialidic, cylindrical to subcylindrical, hyaline, smooth, straight to slightly curved, 12−20 × 1.7−2.7 μm, conspicuous collaretes and periclinal thickenings. – Conidia aseptate, hyaline, smooth, cylindrical to ellipsoidal, 6.8−7.8 × 2.0−2.7 μm (n = 30, = 7.2 × 2.4 μm), with a slightly curved sides, rounded at both ends.

Colonies generated abundant white to pale grey aerial mycelium on PDA. Sporodochia formed on the surface of the medium covered with plenty of slimy olivaceous green conidia. Cultures pale luteous in reverse.

Etymology. – The name reflects the morphology of conidia and its resemblance to P. roridum.

Habitat and distribution. – Agricultural soil, China.

Holotype. – CHINA. Guizhou Province, Guiyang City, Huaxi District, Yangguang county, agricultural garden, from soil, 10 August 2016, leg. Y. Chen (Holotype, GUCC 201608S01; sequences: KY196197 (cmdA), KY126422 (ITS), KY196213 (LSU), KY126429 (tef1), KY196205 (tub2). (living culture HGUP 2016-8006).

Notes. – Paramyrothecium sp. HGUP 2016-8006 was placed amongst P. breviseta, P. foliicola, P. guiyancola and P. roridum with strong bootstrap support (100%(MLBS)/100%(MPBS)/1.0(BI PP)). However, by phylogenetic analyses (Fig 7), its morphological characters are distinctive, with 2−8 conidiogenous cells arisen at ends of conidiophores. Taking the above special characters into account, we considered identifying Paramyrothecium sp. HGUP 2016-8006 as a new species Paramyrothecium verruridum.

We discriminated Paramyrothecium from among 16 Myrothecium-like genera, mostly based on morphological characteristics. Thus, this genus has a special shape and can be recognised easily. We also attempted to use a dichotomous key to classify Paramyrothecium. However, we found that some of the species could not be discriminated by morphology (P. foeniculicola vs. P. parvum; P. tellicola vs. P. terrestris; P. cupuliforme vs. P. humicola; P. verruridum vs. P. guiyangense; and P. roridum vs. P. nigrum) so, it is the better way that a combined phylogeny for the taxonomy of Paramyrothecium species is used presently (Fig. 7).

The following is a list of main morphologic characteristics of each Myrothecium-like genus: 1) Albifimbria – Conidia ellipsoidal to fusiform to limoni-form to subglobose, sometimes bearing a funnel-shaped mucoid apical appendage and straight to its circinate setae (Lombard et al. 2016); 2) Capitofimbria – Olivaceous brown conidia conspicuously verrucose at both ends and shortage of a white setose fringe (Lombard et al. 2016); 3) Dimorphiseta – Formed fusiform, with a funnel-shaped mucoid apical appendage and two types of setae (Type I, flexuous to circinate, verrucose, tapering to an obtuse apex; Type II, tapering to a sharp apex) (Lombard et al. 2016); 4) Gregatothecium – Special character of the slimy olivaceous green conidial masses grown on the conidiophores and sporodochia (Lombard et al. 2016); 5) Inaequalispora – Conidia fusiform to ellipsoidal to asymmetrically ellipsoidal with a slightly curved acute apex and a narrow truncate base with a funnel-shaped mucoid apical appendage and setae tapering to an obtuse apex that becomes lightly verrucose (Lombard et al. 2016); 6) Myrothecium – Conidia ellipsoidal to obovoid, < 5 μm in length (Tulloch 1972); 7) Myxospora – Forming many fusiform conidia, bearing an apical hilum without funnel-shaped mucoid appendages and synnematal conidiomata (Lombard et al. 2016); 8) Neomyrothecium –Similar to Paramyrothecium except that the pulvinate sporodochia lack a white setose fringe (Lombard et al. 2016); 9) Parvothecium – Conidiogenous cells verrucose and conidia asymmetrically ellipsoidal, with slightly curved acute apex and a narrow truncate base (Lombard et al. 2016); 10) Peethambara – Special fusiform conidia morphology with two triangle-shaped marks at its opposite ends and conidiophores gregarious (Subramanian & Bhat 1978a, b, Seifert 1985, Rossman et al. 1999); 11) Smaragdiniseta – Two types of setae (Type I, compacted, verrucose, emerald green; Type II, originating from the marginal hyphae, lightly verrucose, tapering to an obtuse apex, soon growing beyond the length of Type I) (Lombard et al. 2016); 12) Striaticonidium – Synnemata, conidiogenous cells conspicuously curved and conidia black (Lombard et al. 2016); 13) Tangerinosporium – Generating distinctively orange conidial masses on its conidiophores (Lombard et al. 2016); 14) Xenomyrothecium – with oblong-ellipsoidal conidia and lacking setae formed from stroma (Lombard et al. 2016); 15.) Xepicula – Forming an appendage-bearing conidium (Nag Raj 1993).

To our knowledge, these are the first Paramyrothecium species described from China. A more precise taxonomic system of Myrothecium-like was provided by Lombard et al. (2016). Furthermore, the placement of Paramyrothecium is distinctive based on its morphology and phylogenetic analysis in the confusion fungi. Thus, Myrothecium (Tulloch 1972) should be separated into multiple genera to overcome the chaos of having multiple fungal taxonomies (Nag Raj 1993, 1995; Lombard et al. 2016).

Authors: Y. Chen, C. Norphanphoun, J.-Y. Yu, S.-P. Wu, Y. Wang, Y.-M. Wu & Y.-L. Jiang

Pluteus romellii var. luteoalbus Ševčíková & Borovička, var. nov. – Figs. 9–11

Open in a separate window

Fig. 9

Pluteus romellii var. luteoalbus, holotype: a. Basidiospores. b. Basidia., c. Pleurocystidia, d. Cheilocystidia. e. Pileipellis elements. Scale bar 10 μm.

Open in a separate window

Figs. 10–11

Pluteus romellii var. luteoalbus, holotype, basidiocarp, scale bar 1 cm.

MycoBank no.: MB 821253, EMBL-Bank: LT 838190

Diagnosis. – Differs from Pluteus romellii var. romellii by a yellow pileus and a whitish stipe.

Description. – Pileus 14–16 mm broad, convex to plano-convex with indistinct umbo, very slightly rugulose at centre, slightly hygrophanous, glabrous, striate and shortly lacerate at margin, yellow (S00 C00 Y60–S00 C00 Y70), paler towards margin (S00 C00 Y50), distinctly darker at umbo (S10 Y99 M00–Y99 M00–10 C00). – Lamellae free, moderately distant, ± ventricose, whitish to pale yellow (Y10 M00 C00–Y20 M00 C00), later pinkish with concolourous, slightly pubescent edge. – Stipe 60 × 1–2 mm, cylindrical, slightly broadened at base, smooth, finely longitudinally fibrillose, whitish. – Context whitish at a cap, whitish to pale yellow at a stipe part. – Smell and taste indistinct.

Basidiospores 60/3/2 (5.5)6.0–7.5(8) × 5–6.5(7) μm, avl × avw = 6.6 × 5.5 μm, Q = 1–1.35(1.4), avQ = 1.20, broadly ellipsoid, subglobose or globose. – Basidia 15–29(35) × 6–9(10) μm, tetrasterigmate, cylindrical, clavate to subfusiform, colourless. – Pleurocystidia scattered to moderately abundant, (38)45–65(80) × 21–45(55) μm, broadly clavate, broadly cylindrical, broadly utriform to utriform, sometimes with pedicel up to 10(12) μm, rarely ventricose or vesiculose-fusiform, thin-walled, colourless. Lamellar edges sterile, cheilocystidia (18)25–62(72) × 9–31(47) μm, cylindrical, narrowly to broadly clavate, utriform, rarely broadly utriform or broadly cylindrical, thin-walled, colourless. – Pileipellis a euhymeniderm, consists of ventricose, spheropedunculate and broadly clavate elements (20)33–49(60) × (11)15–29(45) μm, with yellow intracellular pigment. – Stipitipellis a cutis of 4–10 (16) μm wide cylindrical colourless hyphae, caulocystidia absent. Clamp connections absent in all tissues.

Etymology. – The epithet luteoalbus refers to yellow pileus and white stipe.

Habitat and distribution. – On fallen mossy stem of undetermined deciduous tree. So far known from only one site in thermophilic Querceto-Carpinetum, South Moravia, Czech Republic.

Holotypus. – CZECH REPUBLIC. Brno-venkov district, Kanice, Zadní Hády Nature reserve, Querceto-Carpinetum, on mossy fallen stem of undetermined deciduous tree. 23 July 2016, leg. P. Ševčík et H. Ševčíková (BRNM 788199).

Material examined (besides holotype). – Pluteus chrysophlebius (orig. P. chrysophaeus): CZECH REPUBLIC. Chynín, Chynínské buky, dead trunk of Fagus, 22 September 1983, leg. J. Baier, det. Herink (BRNM 289486); Čučice, Oslava a Chvojnice, near Oslava river, deciduous woods, 25 October 2016, leg. et det. J. Bět’ák (BRNM 781487); Dolní Věstonice, Děvičky, Acereto-Fraxinetum, 13 July 1955, leg. K. Kříž, det. H. Ševčíková (BRNM 332458); stump of Fraxinus excelsior, 31 August 1955, leg. K. Kříž, det. H. Ševčíková (BRNM 332459); Horní Lomná, Mionší, deciduous woods, 30 August 1962, leg. Novotný, det. H. Ševčíková (BRNM 332444); Kuřim, Šiberná, Querceto-Potentilletum albae, trunk of Quercus, 32 July 1961, leg. F. Šmarda, det. H. Ševčíková (BRNM 332424); Quercetum, 23 October 1954, leg. et det. F. Šmarda 332388); Kuřim, Zlobice, Querceto-Potintilletum albae, Quercus-stump, May 1950, leg. F. Šmarda, det. H. Ševčíková (BRNM 332422); Ulmus-stump, 22 May 1960, leg. F. Šmarda, det. H. Ševčíková (BRNM 332429); Lanžhot, Ranšpurk, dead trunk of Ulmus, 22 June 1967, leg. Lazebníček, det. H. Ševčíková (BRNM 332447); decaying deciduous trunk, 26 June 1992, leg. et det. A. Vágner (BRNM 571060); fallen deciduous trunk, 10 October 1992, leg. et det. A. Vágner (BRNM 571059); dead deciduous trunk, two trees, 24 May 2013, leg. et det. H. Ševčíková (BRNM 747556, 747557); fallen deciduous trunk, 10 October 2013, leg. et det. H. Ševčíková (BRNM 751770); deciduous decaying trunk, 11 April 2014, leg. et det. J. Bět’ák (BRNM 761727); Omice, 15 June 1952, leg. F. Valkoun, det. H. Ševčíková (BRNM 332419); Velká nad Veličkou, Zahrady pod Hájem, dead woods of Carpinus, 27 July 2005, leg. et det. V. Antonín (BRNM 695505); SLOVAKIA. Lehota nad Rimavicou, Dúbrava, Querceto-Carpinetum, deciduous wood, 7 December 2015, leg. J. Hraško, det. H. Ševčíková (BRNM 772178); Snina, Stambočská úboč, fallen trunk of Carpinus, 3 June 2015, leg. J. Pavlík, det. H. Ševčíková (BRNM 781079); HUNGARY. Bátorliget, Fényi erdö, alluvial forest, fallen stem of Quercus robur, 25 October 2006, leg. et det. V. Antonín (BRNM 705010); (Pluteus chrysophlebius - P. luteovirens s. s.): CZECH REPUBLIC. Adamov, Josefovské údolí, Býčí skála, decaying trunk of Carpinus, leg. et det. A. Vágner (BRNM 603145); 17 May 1995, decaying trunk of Fagus, 19 June 2002, leg. et det. A. Vágner (BRNM 670848); fallen trunk of Fagus, 15 September 2011, leg. H. Ševčíková et V. Antonín, det.V. Antonín (BRNM 737386); Josefovské údolí, Slovenská stráň, decaying trunk of Carpinus, 17 September 1986, leg. et det. A.Vágner (BRNM 457767); decaying trunk of Fagus, 18 July 1990, leg. et det. V. Antonín (BRNM 523408); Horní Lomná, Mionší, decaying trunk of Fagus, 29 September 1992, leg. et det. V. Antonín (BRNM 571062); decaying trunk of Fagus, 16 June 2007, leg. L. Hagara, det. V. Antonín (BRNM 706722); Ivaň, Dolní Mušov, floodplain forest, decaying deciduous trunk, 29 June 2001, leg. et det. A. Vágner (BRNM 665359); Kladeruby nad Oslavou, Vlčí kopec, fallen Fagus, 28 September 1994, leg. et det. A. Vágner (BRNM 603007); Lanžhot, Ranšpurk, trunk of Ulmus, 5 October 1988, leg. et det. V. Antonín (BRNM 461736); dead trunk of Ulmus, 28 May 1993, leg. et det. V. Antonín (BRNM 576439); deciduous branch, 27 July 1993, leg. et det. V. Antonín (BRNM 576522); decaying deciduous trunk, two places, 24 May 1995, leg. et det. A.Vágner (BRNM 603178, 603179); Mladeč, NPRVrapač, fallen deciduous trunk, 1 July 1999, leg. et det. A. Vágner (BRNM 648444); Nové Mlýny, Křivé jezero, floodplain forrest, fallen deciduous trunk, 16 June 2005, leg. et det. V. Antonín (BRNM 695486); Pivonice u Pohorské Vsi, Žofínský prales, dead trunk of Fagus, 17 September 2015, leg. et det. H. Ševčíková (BRNM 772294); fallen mosses trunk of Fagus, 17 September 2015, leg. V. Kunca, det. H. Ševčíková (BRNM 772060); Útěchov u Brna, Coufavá, decaying trunk of Fagus, 14 July 1984, leg. et det. A. Vágner (BRNM 457772); 30 September 1984, leg. et det. A. Vágner (BRNM 457765); 14 June 1986, leg. et det. A. Vágner (BRNM 457759);Vilémovice, near Macocha, stump of Fraxinus excelsior, 20 September 1983, leg. D.Vágnerová, det. A.Vágner (BRNM 457758); Vranov, Jelení skok, decaying trunk of Fagus, 21 June 1986, leg. et det. A. Vágner (BRNM 457768); SLOVAKIA. Badín, Badínský prales, decaying trunk of Fagus, 22 August 1995, leg. et det. V. Antonín (BRNM 603576); (Pluteus chrysophlebius – P. galeroides s.s.): CZECH REPUBLIC. Lanžhot, Ranšpurk, fallen trunk of Ulmus, 19 September 1996, leg. V. Antonín, det. H. Ševčíková (BRNM 612145); P. cinereofuscus: CZECH REPUBLIC. Mladá Boleslav, slope, mixed forest of Tilia, Corylus, Ulmus, Carpinus, Quercus, 13 July 2013, leg. S. Tutka, det. H. Ševčíková BRNM 751705; Habrůvka, Habrůvecká bučina, fallen trunk of Fagus, 6 September 2013, leg. et det. H. Ševčíková (BRNM 751723); P. cf. insidiosus: CZECH REPUBLIC. Kulatý dub between Březina and Ochoz u Brna, fallen Quercus, 25 August 2016, leg. et det. H. Ševčíková (BRNM 781263); P. fenzlii: HUNGARY. Vas, Hosszúpereszteg, on heap of sawdust, 7 October 1990, leg. P. Serediuk, det. J. Borovička (BP 88763 originally as P. variabilicolor); P. leoninus: CZECH REPUBLIC. Brno, Baba, near U boudy, mixed forest, heap of branches of Tilia, Quercus, Corylus, 4 September 2014, leg. J. Hrabáková et H. Ševčíková (BRNM 766775); ibid, another heap of branches, 4 September 2014, leg. J. Hrabáková et H. Ševčíková (BRNM 788272); Pluteus romelliivar. romellii: CZECH REPUBLIC. Bílovice nad Svitavou, Anenské údolí, near path, mulch, 2 June 2015, leg. et det. M. Tomšovský (BRNM 771934); Brno, Líšeň, brushwood with Populus tremulae under Hády, deciduous branch, 6 June 2013, leg. F. Fejta, det. H. Ševčíková (BRNM 747550); Brno, Hády, deciduous forest near quarry, decayed deciduous trunk, 21 September 2013, M. Fejta, det. H. Ševčíková (BRNM 751776); Brno, between Hády and U Brněnky, on soil near deciduous twigs, 5 September 2013, leg. et det. H. Ševčíková BRNM (751889); Brno, Hádecká planinka, Querceto-Carpinetum, decayed deciduous woods, 13 May 2010, leg. et det. V. Antonín (BRNM 724855); decayed trunk of Quercus petraea, 11 August 2010, leg. et det. A. Vágner (BRNM 733091); Brno, Kohoutovický potok valley, decayed deciduous woods, 11 October 2001, leg. et det. Z. Bieberová (BRNM 728377); Brno, Bystrc, Rakovec valley, decayed woods of Carpinus betulus, 7 September 2012, leg. et det. V. Antonín et H. Ševčíková (BRNM 745972); Habrůvka, Habrůvecká bučina, near decayed trunk of Fagus, 8 October 2014, leg. et det. V. Antonín et H. Ševčíková (BRNM 767027); Hodonín, Doubrava, mixed forest on sandy soil, 20 June 1984, leg. et det. V. Antonín et A.Vágner (BRNM 305513); Kanice, U Brněnky, Querceto-Carpinetum, deciduous trunk, 5 September 2013, leg. et det. H. Ševčíková (BRNM 751731); Kosmonosy, obora, Carpinus betulus, 26 August 2013, leg. et det. S. Tutka (BRNM 761736); Lanžhot, Cahnov, trunk of Quercus, 11 April 2014, leg. et det. H. Ševčíková (BRNM 761731); Louka u Ostrohu, Háj u Louky, Querceto-Carpinetum; on soil, 2 June 2013, leg. et det. R. Maňák (BRNM 766577); Machnín, Hamrštejn u Liberce, deciduous forest, on soil, 25 August 2013, leg. S. Tutka, det. H. Ševčíková (BRNM 761735), ibid, under Fagus, (BRNM 767004); Mokrá, Mokerský les, Querceto-Carpinetum, decaying deciduous woods, 10 April 2001, leg. et det. A. Vágner (BRNM 665216); near quarry, on soil, 20 September 2013, leg. et det. H. Ševčíková (BRNM 751772); ibid, on soil, 19 November 2014, leg. et det. H. Ševčíková (BRNM 762111); Mokrá u Brna, Sivický les, on soil under Picea abies, Quercus, Rubus, 14 June 2006, leg. et det. A. Vágner (BRNM 699773); decaying deciduous branch, 17 August 2006, leg. et det. A. Vágner (BRNM 705298); Nevojice, Malhotky, decaying trunk of Quercus, 26 September 1997, leg. et det. Z. Bieberová (BRNM 728340); Ochoz u Brna, Údolí Říčky, under Carpinus betulus, 11 November 2001, leg. et det. A.Vágner (BRNM 667875); ibid, Zadní Hády, Querceto-Carpinetum, on soil, 31 May 1981, leg. et det. A. Vágner (BRNM 457750); ibid, decaying trunk Quercus, 22 June 2016, leg. et det. H. Ševčíková (BRNM 781181); ibid, Lysá hora, Údolí Říčky, decaying branch of Corylus avellana, 15 May 2014, leg. et det. H. Ševčíková (BRNM 761858); Prodašice, branch of Populus tremula, 7 September 2014, leg. et det. S. Tutka (BRNM 771985); Veselí nad Moravou, Hutník, mulch of Populus nigra, 1 November 2012, leg. et det. R. Maňák (BRNM 761840); Vápenný Podol, quarry, twig of Quercus, 1 June 2014, leg. et det. R. Doležal (BRNM 767024); SLOVAKIA. Ružomberok-Černová, Černovské lúky (Zrazy), pasture with grove of Corylus and Crataegus, fallen deciduous trunk, 3 April 2014, leg. et det. H. Ševčíková (BRNM 761711); ibid, deciduous twig (BRNM 761712); ROMANIA. Baile Tusnad, Muntii Harghita, in rupe “Stinca Soimilor” ad occidentem ab oppido Baile Tusnad versus, Fagus wood, 2 July 1986, leg. et det. V. Antonín (BRNM 398129); P. variabilicolor: REPUBLIC OF KOREA. Taean Peninsula, Deoksung, Sudeoksa Monastery, mixed forest, dead trunk of Castanea, 7 August 2014, leg. V. Antonín, det. H. Ševčíková (BRNM 788273); SLOVAKIA. Snina, Hrb, heap of sawdust, 10 May 2010, leg. J. Pavlík, det. H. Ševčíková (BRNM 788274).

Notes. – According to our analysis, P. romellii (including the new variety) is a sister species to P. aurantiorugosus which was also demonstrated in the molecular study by Justo & al. (2011). As indicated by tree topology (Fig. 12), P. romellii shows some sequence divergence. The sequence of P. romellii var. luteoalbus completely matches the sequence FJ774073, belonging to a collection of P. romellii from Russia.

Open in a separate window

Fig. 12

Phylogenetic placement of Pluteus romellii var. luteoalbus within closely related Pluteus species inferred from a maximum likelihood analysis of the ITS rDNA molecular data using MEGA7.

Pluteus romellii var. luteoalbus is characterized by a yellow pileus, a whitish stipe, a pileipellis in the form of a euhymeniderm and broadly clavate, broadly cylindrical, broadly utriform to utriform pleurocystidia. Cheilocystidia are narrowly to broadly clavate, utriform to cylindrical. Phylogenetically and microscopically, P. romellii var. luteoalbus is identical to P. romellii var. romellii. Macroscopically, the yellow pileus and whitish stipe are significant for differentiating of this new variety because P. romellii var. romellii has a brown pileus and a yellow stipe. Pluteus romellii f. albidus (Britzelm.) Ferisin is an albinotic form of P. romellii, it has a white pileus and stipe, and a whitish context. Pluteus romellii is commonly found on soil, wood-chips or decayed wood. Pluteus romellii f. albidus is known to grow on fallen stem of a deciduous tree. Pluteus romellii var. luteoalbus was found on decayed deciduous tree, but its growth on soil or on woodchips cannot be excluded.

Pluteus chrysophlebius (Berk. & M.A. Curtis) Sacc. is macroscopically very similar to P. romellii var. luteoalbus. In European literature, species with a yellow pileus and a pileipellis composed of spheropedunculate elements is known as P. chrysophaeus (Schaeff.) Quél., but Justo & al. (2011: 469) selected a lectotype for this species having the brown-pileus. Orton (1986) distinguished three taxa within the P. chrysophaeus: P. luteovirens Rea, P. galeroides P.D. Orton, and P. xanthophaeus P.D. Orton. Pluteus luteovirens, with a lemon-yellow pileus (when it has not greenish-yellow tinge) and a whitish stipe and flesh, is macroscopically the most similar to P. romellii var. luteoalbus. However, Vellinga & Schreurs (1985) synonymized these three taxa under P. chrysophaeus. Additionally, Justo & al. (2011) recently pointed out the similarity of the American P. chrysophlebius (Berk. & M.A. Curtis) Sacc. and the European P. chrysophaeus sensu Vellinga & Schreurs; for the final solution further studies are therefore necessary. With regard to our new variety of P. romellii, taxa of P. chrysophlebius complex have mostly lageniform to fusiform or broadly fusiform pleurocystidia. Broadly clavate, broadly cylindrical to (broadly) utriform pleurocystidia of P. romellii thus represent a significant microscopical difference.

The phylogenetically closest species to P. romellii is P. aurantiorugosus (Trog) Sacc. Macroscopically, colours of the pileus have red or orange tinges and the stipe sometimes has yellow or orange tinges. Microscopically, pleurocystidia of P. aurantiorugosus are mostly vesiculose-fusiform to clavate-fusiform.

Further species with yellow pilei are P. leoninus (Schaeff.) P. Kumm., P. fenzlii (Schulzer) Corriol & P.-A. Moreau, and P. variabilicolor Babos. However, pileipellis of these taxa is not composed of spheropedunculate elements and also differ by the cystidia shape. Moreover, Pluteus fenzlii is macroscopically different by having annulus on stipe and P. variabilicolor by dark floccules on stipe.

Authors: H. Ševčíková, T. Konvalinková & J. Borovička

Sclerostagonospora elegiae Marinc., M.J. Wingf. & Crous, sp. nov. – Figs. 13F, 15A–C, 16A

Open in a separate window

Fig. 13

Line drawings of the conidia of Sclerostagonospora spp. from previous photos and line drawings. A. S. leucadendri (Crous & Palm 1999). B. S. salsolae (Schwarczinger et al. 2000). C. S. centaureae (Sameva & Vanev 1998). D. S. heraclei (Sutton 1980). E. S. pinguis (this study). F. S. elegiae (this study). G. S. sessilis (Gallego et al. 1986a). H. S. sabaleos (Gallego et al. 1986b). I. S. fusiformis (this study). J. S. donacis (Gallego et al. 1986a). K. S. opuntiae (Huhndorf 1992). L. S. phragmiticola (Quaedvlieq et al. 2013). M. S. sulcata (this study). N. S. cycadis (Crous et al. 2011, 2015). O. S. ericae (Crous et al. 2016). Scale bar 10 μm.

Open in a separate window

Fig. 15

Microscopic images of Sclerostagonospora species from restios. A–C. S. elegiae, D–F. S. fusiformis, G–I. S. pinguis, J–L. S. sulcata. A, D, G. J. Vertical section of conidioma, B, E, H, K. Conidiogenous cells, C, F, I, L. Conidia (in bright field). Scale bars: A, D, G, J 50 μm; B, C, E, F, H, I, K, L 10 μm.

Open in a separate window

Fig. 16

Cultures with optimum growth on 2 % MEA (upper) and on OA (lower) in the dark in 13 days at 20 °C except for the ones indicated as 25 °C. A. S. elegiae, B. S. fusiformis, C. S. pinguis, D. S. sulcata. Scale bar 20 mm.

MycoBank no.: MB 823475

Description. – Conidiomata in planta pycnidial, scattered or gregarious, subepidermal, remaining immersed, with the tip of the ostiolar papilla reaching the surface, in vertical section subglobose to globose, 190–240 × 170–215 μm; peridium pseudoparenchymatous, consisting of a few layers of brown, moderately thick-walled, compressed cells (less compressed towards the apex), 8–11 μm thick, inside lined with a layer of hyaline fertile cells. – Conidiogenous cells blastic, hyaline, discrete, doliiform, 4–6 × 2.5–3 μm. – Conidia brown, fusiform to cylindrical, the apex pointed to rounded, the base truncated, (12.5)14–15(17) × (4)5(6) μm, smooth, 3-septate.

Etymology. – Named after the host genus, Elegia.

Culture characteristics. – Colonies on 2 % malt extract agar (MEA) at 20 °C showing optimum growth reaching 37.7 mm in 13 days in the dark, mycelium cottony with raised elevation, showing circular growth with smooth to undulate margin, above greenish grey becoming paler towards the edge, reverse olivaceous, no growth at 30 °C and 35 °C, on oatmeal agar (OA) at 20 °C showing optimum growth reaching 53 mm, above greenish grey to iron grey, reverse greenish black, on potato dextrose agar (PDA) at 20 °C showing optimum growth reaching 44 mm, mycelium cottony with velvety inner circle, above smoky grey with iron grey, reverse olivaceous mouse grey with paler edge, sterile on MEA, OA, PDA, and CLA (carnation leaf agar).

Material examined. – SOUTH AFRICA. Western Cape province, Kirstenbosch National Botanical Garden, culm litter of Elegia equisetacea, 3 December 2001, leg. S. Marincowitz, SL937, holotype, PREM 58934, living culture ex-holotype CBS 118142 = CMW 18281.

Notes. – Sclerostagonospora is characterized by pycnidial conidiomata, holoblastic conidiogenous cells, and pigmented conidia. Conidial morphology of known species is diverse, from elongated to short, smooth to verruculose or striated, lightly pigmented to dark pigmented, and 1- to multi-septation (Fig.13).

Sclerostagonospora Höhn. was treated as a synonym of Stagonospora until Sutton (1980) separated them based on the pigmented conidia of the former. At the same time he relocated some of nomen rejiciendum Hendersonia into the genus. A total of 12 species are currently registered in MycoBank under Sclerostagonospora, of which six are relocated from Hendersonia, namely, S. sabaleos, S. donacis, S. heraclei, S. opuntiae, S. salsolae, and S. sessilis (Höhnel 1917, Gallego et al. 1986a, b, Huhndorf 1992, Schwarczinger et al. 2000). So far only three species, S. ericae, S. cycadis and S. phragmiticola, are provided with DNA sequence data and living cultures (Crous et al. 2011, 2015, 2016; Quaedvlieg et al. 2013).

Two sexual morphs Leptosphaeria leucadendri (Pleosporales) and Montagnula opuntiae (Pleosporales) are linked to Sclerostagonospora (Crous & Palm 1999, Huhndorf 1992). A recent study based on sequences of partial 28S rRNA and rpb2 genes showed that a Sclerostagonospora, S. phragmiticola, resides within the Phaeosphaeriaceae (Pleosporales), suggesting a sexual morph of the genus is phaeosphaeria-like (Quaedvlieg et al. 2013). However, the lack of living culture or DNA sequence data of the type species, S. heraclei, makes the concept of the genus and its status in the family unsettled (Crous et al. 2016).

Several fungi were isolated from the culm litter of Restionaceae (Poales) in South Africa, of which morphological features resemble those of Sclerostagonospora. Phylogenetic analysis based on ITS sequence data, with recently introduced species and those in GenBank, confirmed these isolates are the members of Sclerostagonospora (Fig. 14). Based on both morphological and molecular studies, four species have been identified and proposed as new species in this study.

Open in a separate window

Fig. 14

Phylogram obtained from Maximum likelihood (ML) analyses of ITS region using RA×ML V8.2.4 on the CIPRES Science Gateway v3.3 (Stamatakis 2014). Novel sequences are printed in bold type. ML bootstrap support values (1000 replicates) above 70 % are indicated at the nodes. Scale bar = total nucleotide difference between taxa. The tree was rooted to Phaeosphaeria herbarum and Cochliobolus heterostrophus.

Sclerostagonospora elegiae can be distinguished morphologically by its smooth and brown conidia (Figs. 13F, 15C). Phylogenetic analysis (Fig. 14) using ITS sequence data indicated that CBS 118142 clusters distinctly from the other known species. This isolate differs from its closest phylogenetic neighbour S. pinguis (CBS 118146) by 23 bp in ITS.

Sclerostagonospora fusiformis Marinc., M.J. Wingf. & Crous, sp. nov. – Figs. 13I, 15D–F, 16B

MycoBank no.: MB 823476

Description. – Conidiomata in planta pycnidial, scattered, superficial with the base firmly attached to the substrate, in vertical section subglobose with an ostiolar pore, 110–130 × 115–135 μm; peridium pseudoparenchymatous, 19–20 μm thick throughout the conidioma except for the basal region attached to the substrate 10–15 μm thick, consisting of a few layers of pale brown, moderately thick-walled cells outlined with a few layers of brown, thick-walled cells, the base lack outer layer, the innermost layer consisted of a hyaline, fertile cells. – Conidiogenous cells blastic, hyaline, discrete, ampulliform, 3–4.5 × 3–5 μm. – Conidia yellow to pale brown, fusiform, straight or slightly curved, the apex pointed to rounded, the base truncated, (11)13(15) × (3)3.5–4 μm, 1–3-septate, smooth.

Etymology. – Name refers to the fusiform shape of conidia.

Culture characteristics. – Colonies on 2 % MEA at 25 °C showing optimum growth reaching 15 mm in 13 days in the dark, mycelium velvety, with slightly raised elevation, showing circular growth with smooth margin, aerial mycelium dense, above hazel, reverse olivaceous, no growth at 30 °C and 35 °C, on OA at 25 °C showing optimum growth reaching 23.5 mm, above and reverse greenish grey, on PDA at 25 °C showing optimum growth reaching 19.3 mm, above and reverse saffron, sterile on MEA, OA, and PDA, fertile on CLA.

Material examined. – SOUTH AFRICA. Western Cape province, Kirstenbosch National Botanical Garden, culm litter of Thamnochortus spicigerus, 3 December 2001, leg. S. Marincowitz, SL917, holotype, PREM 58933, living culture ex-holotype CBS 118152 = CMW 18025.

Notes. – Sclerostagonospora fusiformis has smooth, pale brown and fusiform conidia which distinguishes it from the known species (Figs. 13I, 15F). Due to the lack of specimens a limited number of fruiting structures in host tissue were measured.

Phylogenetic analysis (Fig. 14) using ITS sequence data indicated that CBS 118152 clusters distinctly from the other known species. This isolate differs from its closest phylogenetic neighbour S. ericae (CBS 141318) by 6 bp in ITS.

Sclerostagonospora pinguis Marinc., M.J. Wingf. & Crous, sp. nov. – Figs. 13E, 15G–I, 16C

MycoBank no.: MB 823477

Description. – Conidiomata in planta pycnidial, scattered or gregarious, subepidermal, remaining immersed with the tip of the ostiolar papilla reaching the surface or becoming superficial by eroding of the host tissue, in vertical section subglobose, 85–145 × 95–240 μm; peridium pseudoparenchymatous, uneven in thickness, around the ostiole 15–22 μm thick, consisting of several layers of brown, thick-walled cells, lateral tissue 12–16 μm thick, consisting of a few layers of brown, moderately thick-walled, compressed cells, at the base 5–10 μm thick, a few layers of hyaline to pale brown, moderately thick-walled, highly compressed cells, lined with a layer of hyaline, fertile cells inside. – Conidiogenous cells blastic, hyaline, discrete, ampulliform, 3.5–10 × 2–4 μm. – Conidia subhyaline to pale brown at young, becoming dark brown when mature, fusiform to ellipsoidal, often asymmetrical, the apex pointed, the base truncated, (16)18.5–19(21.5) × (6)8–9 μm (avg. 18.9 × 9 μm), 3-septate, verruculose, thick-walled.

Etymology. – Name refers to a plump feature of conidia.

Culture characteristics. – Colonies on 2 % MEA at 20 °C showing optimum growth reaching 44 mm in 13 days in the dark, mycelium cottony with slightly raised elevation, showing circular growth with smooth margin, above greenish grey becoming paler towards the edge, often with white patches, reverse olivaceous, no growth at 30 °C and 35 °C, on OA at 20 °C showing optimum growth reaching 52 mm, colony features similar to that on MEA, on PDA at 20 °C showing optimum growth reaching 41.8 mm, above pale purplish grey, reverse greenish black with paler margin, sterile on all MEA, OA, PDA, and CLA.

Material examined. – SOUTH AFRICA. Western Cape province, Jonkershoek Nature Reserve, culm litter of Cannomois virgata, 15 June 2001, leg. S. Marincowitz, SL744, holotype, PREM 58931, living culture ex-holotype CBS 118146 = CMW 17948; culm litter of Ischyrolepis cf. gaudichaudiana, 31 July 2001, leg. S. Marincowitz, SL881, PREM 58932.

Notes. – Sclerostagonospora pinguis is distinguished by its brown, verruculose, and thick-walled conidia (Figs. 13E, 15I).

Phylogenetic analysis (Fig. 14) using ITS sequence data indicated that CBS 118146 clusters distinctly from the other known species. This isolate differs from its closest phylogenetic neighbour S. elegiae (CBS 118142) by 23 bp in ITS.

Sclerostagonospora sulcata Marinc., M.J. Wingf. & Crous, sp. nov. – Figs. 13M, 15J–L, 16D

MycoBank no.: MB 823478

Description. – Conidiomata in planta pycnidial, scattered or gregarious, subepidermal, remaining immersed with the tip of the ostiolar papilla or pore reaching the surface, in vertical section globose to ovoid, occasionally loculated, 60–100 × 50–90 μm; peridium pseudoparenchymatous, uneven in thickness, around the ostiole 5.5–10 μm thick, consisting of a few layers of dark brown, thick-walled cells, lateral and basal tissue 8–9 μm thick, consisting of a few layers of hyaline to slightly pigmented, moderately thick-walled cells, the inner-most layer composed of a layer of hyaline, fertile cells. – Conidiogenous cells blastic, hyaline, discrete, cylindrical to doliiform, 3–10 × 2.5–4 μm. – Conidia brown, cylindrical, straight or slightly curved, the apex rounded, tapering to the truncate base with minute marginal frills, (13)14–15(17) × (4)5–6(7) μm, 3-septate, slightly constricted at each septum, vertically furrowed, thick-walled.

Etymology. – Name refers to a furrowed surface of conidia.

Culture characteristics. – Colonies on 2 % MEA at 25 °C showing optimum growth reaching 17.2 mm in 13 days in the dark, mycelium velvety, showing circular growth with smooth to undulate edge, aerial mycelium dense, above honey to dark brown with white edge, reverse sepia, no growth at 35 °C, on OA at 20 °C showing optimum growth reaching 50.8 mm, mycelium woolly, flat, aerial mycelium medium sparse, above smoky grey to grey olivaceous, reverse sepia, on PDA at 20 °C showing optimum growth reaching 45.8 mm, above saffron with ochreous patches, reverse sepia with paler edge, slight pigmentation on medium on MEA and on PDA at 25 °C, sterile on MEA, OA, PDA, and CLA.

Material examined. – SOUTH AFRICA. Western Cape province, Kirstenbosch National Botanical Garden, culm litter of Ischyrolepis subverticellata, 3 December 2001, leg. S. Marincowitz, SL1101, holotype, PREM 58935, living culture ex-holotype, CBS 118224 = CMW 18063.

Notes. – Sclerostagonospora sulcata has furrowed ornamentation in conidia, which reminds of S. opuntiae, however differs in its truncated base with minute marginal frill (Figs. 13M, 15L). This fungus shows better growth on OA and PDA (45–50 mm) than on MEA (17 mm).

Phylogenetic analysis (Fig. 14) data indicated that CBS 118224 is identical with S. fusiformis (CBS 118152) in ITS sequence. However, CBS 118224 is introduced as a new species based on morphological features. CBS 118224 has brown, cylindrical and sulcate conidia, whereas S. fusiformis has pale brown, fusiform and smooth conidia.

The same isolates representing four new Sclerostagonospora spp. in this study were also examined by Crous et al. (2012). They sequenced the ITS and LSU regions of the isolates and the sequence data were identical to the ones produced in this study, except for S. sulcata (CBS 118224) of which sequences showed some discrepancy in both gene regions. The reason of this discrepancy needs a further investigation.

Authors: S. Marincowitz, F. Jami, M.J. Wingfield & P.W. Crous

Interesting taxonomical notes, host and geographical records

Cortinarius scaurocaninus Chevassut & Rob. Henry, Doc. Mycol. 12 (no. 47): 25 (1982). – Fig. 17

Open in a separate window

Fig. 17

Cortinarius scaurocaninus, WU 13419. a. Basidiomata, phot. W. Klofac. b. Spores, scale bar 5 μm.

Synonyms. – Cortinarius glaucopus var. olivaceus f. ingratus Moënne-Locc., Cortinarius olidovolvatus Bon &Trescol s. auct.

Material examined . – AUSTRIA. Styria, Bad Gleichenberg, Kurpark, near Quercus (Fagus, Carpinus), 22 October 1994, leg. W. & S. Klofac, WU 13419, Genbank no. MG489871. The investigated collection was characterized by a strongly innately fibrillose pileus, absence of blue-violet colours, a stipe base with a marginate bulb, abundant veil at the bulb margin, dark brown reaction of potassium hydroxyde on pileus surface and stipe base and Lugol negative with NH₃.

Notes. – In the course of the Austrian Barcode Of Life project, ABOL (https://www.abol.ac.at/en/), specimens of the genus Cortinarius were sequenced. It turned out that the ITS sequence of a collection, morphologically determined as C. glaucopus var. acyaneus, because of lack of blueish colours, matched 100 % with the C. scaurocaninus sequence KF732641 (Limatainen et al. 2014). Cortinarius scaurocaninus was described from the Mediterranean area and is a member of the complicated C. glaucopus alliance. It is characterized by ample veil on young basidiomata, very prominently innately fibrillose pileus surface, only very young bluish lamellae and bluish hues on the stipe and in the stipe context, and brown reaction of potassium hydroxyde on pileus surface (Schmidt-Stohn et al. 2016). Ecologically, it forms ectomycorrhizae with Quercus species. It occurs in warm-toned, Central European and Mediterranean deciduous forests over limestone and silicate, mostly near Quercus and Carpinus, once also near Fagus. So far collections are known from Germany, France, Italy, Spain and Hungary (Schmidt-Stohn et al. 2016). This is the first record of this species for Austria.

Author: I. Krisai-Greilhuber

Humicola grisea Traaen, Nytt Mag. Natur. 52: 34 (1914). – Fig. 18

Open in a separate window

Fig. 18

Humicola grisea (CCCT 17.02). A, B. Fertile hyphae, conidiogenous cells and conidia. C. Conidia. Scale bars 10 μm. The arrows in B and C show conidial germ pores.

Description. – Vegetative hyphae septate, branched, straight to flexuous, hyaline to light olivaceous brown, smooth to verruculose, thin-walled, 1.5–4.5 μm wide, often with thick, dark septa. Conidiogenous cells mono- to polyblastic, scattered or forming more or less dense clusters, arising laterally at right angles from undifferentiated hyphae, but sometimes terminal or intercalary, mostly subcylindrical to clavate, light olivaceous to dark olivaceous brown, smooth to verruculose, thin-to thick-walled, 1–18 × 2–7 μm. Conidia almost always solitary, very rarely forming chains of 2–3, dry, globose to dacryoid, aseptate, pale olivaceous brown to dark brown, smooth to inconspicuously verruculose, 9–14(17.5) × 8–13.5(15.5) μm, with 1(2) conspicuous germ pore(s) 2–4 μm wide.

Culture characteristics. – Colonies after 7 d at 25 °C attaining 41 mm. diam. on MEA and 37 mm. on PDA. On MEA velvety, dark grey to black at the centre, with abundant drops of a colourless exudate, light grey to whitish towards the periphery, with a slightly lobulated margin; reverse black with a light grey margin. On PDA velvety to cottony, umbonate, dark grey at the centre, light brownish grey to light grey at the periphery, with a fimbriate margin; reverse black with a light brownish grey to light grey margin.

Material examined. – CHILE, Región Metropolitana, Santiago, from decaying bark of unidentified Arecaceae on soil, 27 April 2015, leg. H. Madrid (culture CCCT 17.02).

Notes. – During a taxonomic study of saprobic fungi in central Chile, an interesting dematiaceous hyphomycete was isolated from decaying palm bark (CCCT 17.02). The fungus produced abundant whitish mycelium on the natural substrate after 3 weeks of incubation in a moist chamber at room temperature. This strain was originally identified as “Gilmaniella sp.” on account of its mono- to polyblastic cells giving rise to one-celled, mostly globoid, melanised conidia with conspicuous germ pores (Barron 1964, Seifert et al. 2011). Conidial dimensions were clearly different from those of the currently accepted species (Barron 1964, Moustafa 1975, Sivanesan & Sutton 1985, Moustafa & Ezz-Eldin 1989, Umali et al. 1998, Dubey et al. 2011, Goh et al. 2013) and this initially led us to suspect that it was a novel taxon.

A BLAST search, however, revealed that the ITS sequence of CCCT 17.02 (GenBank KU705826) was 100 % identical to that of an authentic, probably ex-type strain of Humicola grisea, CBS 119.14 (GenBank AY706334). This finding was surprising, since the presence of conidial germ pores has been considered as the main morphological feature separating the genus Gilmaniella from Humicola (Ellis 1971, Seifert et al. 2011). Other close matches to the Chilean strain were Chaetomium vitellinum CBS 250.85 (GenBank JX280859, 99 % identical), C. crispatum CBS 408.81 (GenBank JX280781, 99 % identical) and Thielavia antarctica CBS 123565 (GenBank KM655343, 98 % identical), indicating affinities to the Chaetomiaceae, where the type species of Humicola, H. fuscoatra, resides (Wang et al. 2016). We also attempted to generate ITS and LSU sequences from the ex-type strain of Gilmaniella humicola, CBS 220.65, the type species of Gilmaniella. Sequences were not of optimal quality, but BLAST searches with the preliminary ITS and LSU sequences suggested affinities to species traditionally placed in Lasiosphaeriaceae, with the closest matches being members of a phylogenetic group named “clade A” in Cai et al. (2006). This clade includes an assemblage of morphologically diverse species in polyphyletic sexually-typified genera such as Apiosordaria, Cercophora, Podospora and Zopfiella, as well as asexual morphs referred to as Cladorrhinum, humicola-like or chrysosporium-like (Guarro et al. 1991, Cai et al. 2006, Madrid et al. 2011). The definitive phylogenetic placement of G. humicola requires further study, and we will attempt to generate new ITS and LSU sequences for this fungus. An ITS-based tree showing the placement of H. grisea among other conidial and sexual members of Chaetomiaceae has been published in Nonaka et al. (2012), and we consider it unnecessary to include a similar phylogenetic analysis herein.

Micromorphological features of CCCT 17.02 were described from cultures on water agar with sterilized pine needles after 7 d at 25 °C. Colonies on this medium produced very scarce, sparse, mostly immersed mycelium, appearing practically translucent to the naked eye, except for areas with clusters of dark brown conidia. Therefore, we preferred to describe colonies on more nutritive media, i.e. MEA and PDA, where they appeared richer in texture and colour. The morphology of strain CCCT 17.02 (Fig. 18) suggests that the presence or absence of conidial germ pores is not a reliable feature to distinguish the genera Gilmaniella and Humicola. Although H. grisea traditionally has been considered to lack conidial germ pores (Ellis 1971, Seifert et al. 2011), these structures can be observed in microphotographs from the ex-type strain published by White & Downing (1953). Those authors, however, did not mention germ pores in their description of the fungus. The type species of Gilmaniella, G. humicola, produces a conidiogenous apparatus formed by inflated cells (Barron 1964), which has not been described in Humicola. This character might be more helpful in separating the genera Gilmaniella and Humicola, but a detailed study of all species available in culture is necessary to corroborate this hypothesis.

The conidial size described for H. grisea by Traaen (1914) and Ellis (1971), i.e. 12–17 μm diam., closely resembles that of CCCT 17.02 and so the identification of this species is supported by both morphology and DNA sequence data. A phialidic synasexual stage illustrated in Ellis (1971) was rarely observed in culture, but it was absent in the slides used for the species description provided herein. Humicola grisea is a strongly cellulolytic fungus which might deserve further studies due to its potential biotechnological applications (White & Downing 1953). It has a widespread geographical distribution and has been reported from soil, wood and plant debris mainly from regions with cold winters in Europe and North America (White & Downing 1953, Ellis 1971). This taxon, however, has not been mentioned in comprehensive lists of fungal species from Chile (Mujica et al. 1980, Lazo 1996, Minter & Peredo Lopez 2006) and represents an interesting new record.

Authors: H. Madrid, A. Kolecka & T. Boekhout

Opegrapha parasitica (A. Massal.) H. Olivier, Bull. Acad. Intern. Géogr., Bot. 16: 190 (1906) – Fig. 19

Open in a separate window

Fig. 19

Opegrapha parasitica. a. Lirellate apothecia, parasite on Aspicilia sp. Scale bar 1 mm. b.Section through apothecium with obscured hymenium, asci and ascospores. Scale bar 25 μm. c. Cross-section of apothecium with ascus and ascospores. Scale bar 25 μm.

Description. – Lichenicolous. – Apothecia lirellate, short or long, broad, black, with a narrow slit, partly immersed in the thallus of Aspicilia sp., 0.25 × 0.7 mm diam. or 0.25 × 0.25 mm diam. – Hymenium obscured. – Hypothecium colourless. – Asci clavate-cylindrical, 40–50 × 12–14 μm, 4-spored. – Ascospores 17 × 6 μm, hyaline, 4-celled, verrucose (Fig. 19a–c).

Notes. Opegrapha parasitica is a lichenicolous fungus, growing on crustose lichens on limestones (Aspicilia species, Caloplaca cirrochroa, Verrucaria hochstetteri) and on Xanthoria parietina (Hawksworth 1983, Seaward & Pentecost 2001). A detailed description is provided by Smith et al. (2009).

In recent years many noteworthy new records have been added to the lichen flora of Turkey (Aptroot & Yazıcı 2009, Aptroot et al. 2015; for further references see Yazici & Aslan 2016a,b). However, there are many lichenologically unexplored parts needed to be studied in the country.

Twenty taxa of Opegrapha, and 57 taxa of Rinodina have thus far been reported from Turkey. Although more than 1650 lichen taxa have been noted from Turkey (Yazici & Aptroot 2015), only six lichen taxa have thus far been reported from the Muş region (Yazici & Aslan 2016a,b), and 31 lichen species were noted from Bitlis region (Çobanoğlu 2005, Çobanoğlu & Yavuz 2007, Vondrák et al. 2012). Within our projects “Lichen flora of Muş and Bitlis Provinces” we identified two lichens as new records for Turkey. Opegrapha parasitica is one of them. It is known from Europe (England, France, Germany, Greece, Ireland, Norway, Poland, Spain, now also Turkey), Middle East (Syria), New Zealand, North America, and northern Africa (Hawksworth 1983, John et al. 2004, Wirth et al. 2011).

Material examined : TURKEY. Muş: Bulanık, 5 km to Arakonak village, roadside, 39° 04' 17.42" N, 42° 07' 35.93" E, 1605 m, on Aspicilia cf. calcarea, 29 May 2015, leg. et det. K. Yazici, A. Aslan & A. Aptroot. (KTUB–2457). Associated hosts: Lepraria membranacea, L. incana, Solenopsora candicans, Rinodina sp., Lecanora cf. hageni.

Rinodina zwackhiana (Kremp.) Körb., Syst. Lich. Germ. (Breslau): 126 (1855). – Fig. 20

Open in a separate window

Fig. 20

Rinodina zwackhiana. a. Habitus. Scale bard 1 mm. b.Section through apothecium with hypothecium, hymenium and blue gray epihymenium. Scale bar 500 μm. c. Section through apothecium with young ascospores and paraphyses. Scale bar 25 μm. d. Cross-section of apothecium with ascus ascospores, paraphyses and blue-gray epihymenium. Scale bar 25 μm

Description. – Thallus crustose, ± squamulose, sometimes forming ± rosettes, 2–2.5 cm diam., appressed to the substratum, distinctly areolate, light brown, light brown-orange, lobate; lobes short, thick, hump, formed like tiles, surface with white crystals in points, with ellipsoidal tips, undivided or dichotomic, sometimes overlapping, 0.25–0.30 diam.; – soralia absent; – Apothecia aggregated, sessile or sometimes constricted at the base, 1.25–1.30 mm diam., mostly in the centre of the thallus, rare towards the lobes; thalline margin concolorous with the thallus, 0.25 mm diam., entire; exciple ± pigmented; disc 1 mm diam., reddish brown or dark red, ± concave, flat at first, some large ones horizontal, saddle-shaped and rarely slightly convex later, fissured. – Hymenium ± pigmented, 120–140 μm; – epihymenium with blue-grey pigmentation. – Hypothecium hyaline, 120–190 μm, light yellow-brown; asci 8-spored, clavate, 60–90 × 20–25 μm. – Ascospores 2-celled, brown, ellipsoid with rounded lumina, pigmented septa at maturity, Bicincta-type, 15.45–17.20 × 9.8–11.25 μm. – Pycnidia absent. All spot tests negative.

Notes. – Rinodina zwackhiana is a mainly mild-temperate lichen, found on steeply inclined to slightly underhanging calcareous rocks, on walls, and sometimes a juvenile parasite on other lichens, probably more widespread, and much overlooked. A detailed description is provided by Nash et al. (2001). It is known from Asia (China), Europe (Austria, Czech Republic, France, Germany, Italy, Poland, Romania, Slovakia, Slovenia), North Africa (Morocco), and North America (Bates et al. 2010, Nash et al. 2001, Vondrák et al. 2006, Wilfling & Mayrhofer 2002). Rinodina zwackhiana is new to Turkey.

Material examined. – TURKEY. Bitlis: Centre, mainroad of Siirt-Bitlis, 4 km to Bitlis, roadside, 38° 21¢ 36.89² N, 42° 03¢ 51.44² E, 1416 m, on calcareous rock, 08 August 2016, leg. et det. K. Yazici, A. Aslan & A. Aptroot. (KTUB–2459). Associated species: Acarospora cervina, Aspicilia cinerea, Caloplaca saxicola, Collema tenax, Psorotichia shaereri, Verrucaria fuscella, Xanthoria elegans.

Authors: K. Yazici, A. Aptroot & A. Aslan

Xerula strigosa Zhu L. Yang, L. Wang & G. M. Mueller, Acta Bot. Yunnanica 2009. – Figs. 21–22

Open in a separate window

Fig. 21

Xerula strigosa. a. Basidiocarp. b. Pleurocystidia. c. Basidiospores. d. Basidia. Scale bars: a 1 cm; b 30 μm; c 12 μm; d 15 μm.

Open in a separate window

Fig. 22

Phylogenetic analysis of Xerula strigosa collected from Pakistan (■) based on nrITS-rDNA regions. The tree is based on maximum likelihood method using Jukes-Cantor model (Jukes & Cantor 1969). Bootstrap values have been shown on each branch. The analysis involved 50 sequences, with Flammulina velutipes as the outgroup.

Description. – Pileus 20–45 mm diam., convex to plano-convex, light brown to dark brown, fibrillose or fine scaly, umbonate, depressed broadly over disc, central disc yellowish brown, not viscid, margin smooth to slightly dentate; context thin to moderately thick at central disc, soft to firm, white, unchanging when bruised or cut. – Lamellae cream to white, adnate to adnexed, subdistant, edges entire, fleshy, centrally thick, lamellulae alternating with lamellae. – Stipe 3.5–12 × 0.3–0.5 cm (incl. pseudorhiza), yellowish brown, central, cylindrical to sub-cylindrical, base tapering, firm and woody without volva or zones. – Odour mild.

Basidiospores 13.0–18.0 × 11.5–13.5 μm, Q = 1.23 ellipsoidal to broadly ellipsoidal, rarely subglobose, hyaline, pale yellow in 5 % KOH, thin-walled, prominent apiculus 1.9–3 μm, inamyloid, non-dextrinoid. – Basidia 28.5–40.12 × 10.0–13.0 μm, 4–spored, rarely 2-spored, clavate, hyaline, clamp connection not observed, hyaline to olive green granules or droplets in 5 % KOH. – Pleurocystidia 101.5–118.0 × 19.0–26.0 μm, fusiform, ventricose, hyaline to pale yellow, thick-walled, 3–5 μm, with bluntly ended apex, yellow crystals on apices. – Cheilocystida similar to pleurocystidia. – Peillipellis loosely arranged hyphae, light yellowish-brown, pileocystidia 33.5–105.0 × 23.5–33.0 μm, thick-walled, broadly clavate, hyaline.

Material examined. – PAKISTAN. Khyber Pakhtunkhwa, Himalayan Moist Temperate Forests, Khanspur (34.0167° N, 73.4167° E), at 2250 m a.s.l., solitary, on moist ground near a mixed vegetation of Abies pindrow (Royle ex D.Don) Royle, Pinus wallichaina A. B. Jacks. and Quercus incana Bartram, 24 August 2010, leg. A. Razaq (LAH 240806).

Notes. – The Pakistani specimen is in accordance with the holotype description by Wang et al. (2008). The four-spored basidia and the size and shape of the thick-walled pleurocystidia of our collection conform to the holotype but the length of spore varies slightly (Chinese 9.5–16.5 × 8.5–15.5μm vs. Pakistani 13.0–18.0 × 11.5–13.5μm). Ecologically, both collections have been observed in mixed pine forests with deciduous trees like oak (Wang et al. 2008).

The internal transcribed spacers (ITS1 & ITS2) and 5.8S rDNA was sequenced of our specimen using universal primers pairs (ITS1f & ITS4) (White et al. 1990) and showed maximum nucleotide resemblance with X. strigosa sequences (KF530552.1, KF530553.1, KF530555.1) from China as highest nucleotide similarity matches. In phylogenetic analysis, all the sequences of Xerula and its allied genera recovered in three clades: clade I, Xerula, clade II, Strobilurus and Paraxerula (Fig. 22). Our phylogenetic analysis showed that the two latter genera are monophyletic, while the former is poly-phyletic as already noted by Petersen & Hughes (2010). Pakistani specimen recovered in sub-clade I.I (Fig. 22) along with its respective X. strigosa sequences from China. In this subclade, two sequences are noteworthy: X. strigosa (KF530556.1) and X. hispida (AF321485.1). The placement of X. hispida (AF321485.1) from China among the sequences of X. strigosa showed that actually it is X. strigosa while the clustering of X. strigosa (KF530556.1) as a sister clade to all other sequences of X. strigosa indicates either that it has more intraspecific variation or is a misidentification (Fig. 22, clade I.I). Xerula strigosa has been described from China and is now reported first from Pakistan based on morphological and molecular characterizations. Petersen & Nagasawa (2005) also mentioned Pakistani material but without a details.

Authors: A. Razaq, S. Ilyas, S. Shaheen & A.N. Khalid

Acknowledgements

References