An overview of the genus Coprotus (Pezizales, Ascomycota) with notes on the type species and description of C. epithecioides sp. nov. Author Kusan, Ivana Author Matocec, Neven Author Jadan, Margita Author Tkalcec, Zdenko Author Mesic, Armin text MycoKeys 2018 29 15 47 http://dx.doi.org/10.3897/mycokeys.29.22978 journal article http://dx.doi.org/10.3897/mycokeys.29.22978 1314-4049--15 Coprotus epithecioides Matocec & I. Kusan sp. nov. Figs 4, 5 Type . CROATIA. Lika-Senj County, Sjeverni Velebit National Park, northern part of the Mt. Velebit, Hajducki kukovi area, 150 m W from Golubic peak (1650 m), 44°46.05'N ; 15°00.88'E , 1580 m a.s.l.; on dung of chamois ( Rupicapra rupicapra ), 11 Oct 2017, I. Kusan (holotype CNF 2/10450, GenBank sequences ITS MG593539, LSU MG593540). Etymology. The specific epithet refers to epithecium-like ascal protective formation composed of swollen apical paraphyses cells. Description. Apothecia not confluent regularly circular to irregular from the top view, at first oblate, then turbinate, finally pulvinate from the side view, sessile, subhyaline to creamy grey or pale yellowish, glabrous, *170-420 µm in diameter, solitary or gregarious. Hymenium only very finely scurfy, ascal protrusions not clearly visible. Margin rounded in vertical median section, entire and smooth, expanded with downwards positioned rim, never raised above hymenial plane. Outer surface smooth, concolorous with the hymenium. Subicular hyphae indistinguishable. Hymenium *75-98 µm thick. Asci shortly cylindric with slightly truncate apex, *60-74.8 x 13.4-15.5 µm , †51.5-62 x 11.8-14 µm (Q = 3.8-5.2), when mature *protruding above hymenium up to 7.5 µm , pars sporifera*28-34 µm , 8-spored, hyaline; base attenuated, bifurcate, arising from perforated crosiers; only optimally oriented fully mature asci with flat lentiform operculum clearly delimited prior the spore discharge, *6.3-6.6 µm in diam. and *0.5 µm thick, lateral wall 3-layered, *0.6 µm thick, after spore discharge operculum as a rule clearly visible; in IKI inamyloid; in CR outermost wall vividly rutile-red throughout the ascal length, median layer pale rutile-yellow, innermost layer greyish; in CB asci cyanophobic. Ascospores *7.9 -8.8- 9.6 x 4.8 -5.2- 5.6 µm , †8 -9.1- 9.5 x 4.2 -5- 5.2 µm , *Q = 1.5 -1.6- 1.9, †Q = 1.6 -1.9- 2.0, bilaterally symmetrical with one side flattened, subphaseoliform to phaseoliform, poles rounded, 1-celled; uni- to biseriate in living asci, freshly ejected remain in a group for a while due to the delicate subglobose sticky sheath enveloping individual spores; hyaline, smooth; wall 3-layered, 0.4 µm thick, perispore dull, epispore brightly refractive, endospore subhyaline, barely optically differentiated; eguttulate, uninucleate, nucleus always +/- polarly positioned, 2.2-2.5 µm wide; in IKI perispore and epispore not stained, endospore purplish, nucleus slightly contrasted; in CRB without differential stainings, the edges of spore sheath sharply contrasted, after applying KOH spore sheath instantly dissolves, perispore not loosening , endospore layer purplish-rosaceous; in CB with one eccentrically positioned de Bary bubble in mature spores, perispore not loosening, moderately cyanophilic. Paraphyses +/- densely septate, with thin, hyaline walls, cylindric in the lower part, often branched in the upper part, rarely simple, apically +/- bent clavate or capitate, not producing copious exudate ; of two types: (a) epithecioid, reaching higher level, with apical short and capitate cell, *6.8-10 x 5-9.9 µm , †6.2-11.2 x 4-8 µm , with 1-2 subapical cells often also swollen (moniliform), forming +/- continuous layer above living immature asci, and (b) of usual type with elongated clavate apical cells, *8.2-14.8 x 2.3-4.4 µm , †5.5-11 x 2-3.3 µm ; both types may contain yellow-orange pigment, often of crystalloid, fibrillar structure; pigment in IKI cinnamon-grey, in CRB purplish-lilac, often barely visible since mainly included in large globose, deeply stained blue-violet vacuole; in CB wall cyanophobic, cytoplasm pale greyish-blue. Margin reduced, composed of textura globulosa-angularis, cells not elongated, *3.8-6 µm wide, cylindric-elongated cells absent; weakly cyanophilic in CB. Subhymenium hyaline, not differentiated from medullary excipulum. Medullary excipulum hyaline, in the central part *32-56 µm thick, in the middle flank *10-14 µm thick, composed of textura epidermoidea, cells thin-walled, *2.3-4.8 µm wide, in CB cyanophobic. Ectal excipulum hyaline, in the middle flank *17-22 µm thick, composed of textura globulosa-angularis, cells *9.8-16.5 x 7.8-14.7 µm , †4.5-12 x 2.3-9.5 µm , walls thickened, refractive, yellowish, *0.5-0.7 µm thick, in CB cell walls slightly cyanophilic. Overall excipulum without crystalline matter, dextrinoid reaction in MLZ and colouring in KOH; in IKI inamyloid and devoid of glycogene accumulations. Anamorph not found. Figure 4. Coprotus epithecioides (CNF 2/10450, holotype). a Fresh apothecia on Rupicapra rupicapra dung b Cross section through the whole apothecia c Cross section in dark field d Asci e Freshly ejected ascospores glued together with a sheath and individual ascospores f Freshly ejected ascospores in phase contrast g Epithecioid paraphyses h Clavate paraphyses with pigment content i Epithecioid hymenial cover j Excipular flank k Marginal tissue. All elements observed in tap water and in living state, except two asci on d marked with a cross (†); Scale bars: a 0.5 mm, b, c 50 μm , d-k 10 μm , phot. N. Matocec & I. Kusan . Figure 5. Coprotus epithecioides (CNF 2/10450, holotype). a Asci with ascospores containing de Bary bubbles, red markings show opercular delimitation b Paraphyses c Ectal excipulum from top view d Excipular flank f Paraphyses g Ascospores . a-c †CBd†MLZe*CRBf†IKI. Scale bars: a-f 10 μm , phot. N. Matocec & I. Kusan . Distribution and ecology. The species is known so far only from Mt. Velebit, Croatia. The only collection originates from chamois dung in the alpine karstic habitat. Other specimens examined. None. Notes. Coprotus epithecioides has several characters making it distinct from other species in the genus. The paraphyses are of two types, along with the usual filiform-clavate ones, there are also an abundance of those with very short, swollen apical cells, that mutually form an epithecioid protective layer over immature asci, a character not recorded so far in the genus Coprotus . Additionally, in the epithecioid type, 1-2 subapical cells are often also swollen. This gives the paraphyses a moniliform appearance. When present, paraphysal pigments are most often orange to reddish-orange and crystalloid, i.e. of fibrillar shape, resembling the carotenoid pigmentation of Scutellinia species. Spores are highly bilaterally symmetric compared to C. glaucellus , C. subcylindrosporus , C. argenteus and C. sexdecimsporus (which has only inconspicuously and partly bilaterally symmetric spores) and the spores are significantly shorter than those of C. subcylindrosporus , C. argenteus and C. sexdecimsporus . Coprotus glaucellus differs by the presence of only apically uninflated to subclavate paraphyses which do not form an epithecioid protective cover over immature asci. Also it has notably elongated cells at the marginal edge. As elaborated above, paraphysal cytoplasmic pigments normally also develop in this species if the fungus is strongly exposed to sunlight or artificial light with ultraviolet wave-lengths. The pigmentation is completely absent if the apothecia is grown continually under dark or low-light conditions (see notes under C. sexdecimsporus ). Worldwide identification key to the putative species of the genus Coprotus

2
4
C. arduennensis J.R. De Sloover
3
C. marginatus Kimbr., Luck-Allen & Cain
C. dhofarensis Gene , El Shafie & Guarro
textura intricatatextura globulosa-angularis	5
textura globulosa-angularis	6
C. baeosporus Jeng & J.C. Krug
textura globulosa-angularis	C. ochraceus ss. Thind et al. ( Thind et al. 1978 )
texura globulosa-angularis	7
textura globulosa-angularis	11
8
9
C. uncinatus Yei Z. Wang
C. tetrasporus Haeffner , nom. inval.
C. epithecioides Matocec & I. Kusan
10
C. granuliformis (P. Crouan & H. Crouan) Kimbr.
C. trichosuri A.E. Bell & Kimbr.
12
17
13
14
C. sexdecimsporus (P. Crouan & H. Crouan) Kimbr. & Korf
C. duplus Kimbr., Luck-Allen & Cain
15
16
C. rhyparobioides (Heimerl) Kimbr.
C. albidus (Boud.) Kimbr.
*†	C. niveus (Fuckel) Kimbr., Luck-Allen & Cain
C. winteri (Marchal & E .J. Marchal) Kimbr.
18
19
C. sarangpurensis K.S. Thind & S.C. Kaushal
C. disculus Kimbr., Luck-Allen & Cain
20
29
C. subcylindrosporus J. Moravec
21
textura globulosa-angularis	C. ochraceus (P. Crouan & H. Crouan) J. Moravec
22
C. vicinus (Boud.) Kimbr., Luck-Allen & Cain
23
24
27
C. aurora (P. Crouan & H. Crouan) K.S. Thind & Waraitch
25
Ascophanus aurantiacus Velen.
26
C. bilobus (Velen.) J. Moravec
C. breviascus (Velen.) Kimbr., Luck-Allen & Cain
C. breviascus ss. Dokmetzian et al. ( Dokmetzian et al. 2005 )
28
C. luteus Kimbr.
C. aff. luteus (cf. Doveri 2004 )
or	30
32
C . argenteus (Curr.) Waraitch
31
C. dextrinoideus Kimbr., Luck-Allen & Cain
C. leucopocillum Kimbr., Luck-Allen & Cain
Trichosurus vulpecula	C. trichosuri A.E. Bell & Kimbr.
33
C. sphaerosporus J.L. Gibson & Kimbr.
34
C. glaucellus (Rehm) Kimbr.
C. lacteus (Cooke & W. Phillips) Kimbr., Luck-Allen & Cain

Table 2. Coprotus species overview - macroscopy and ecology.

Species	Apothecial shape	Apothecial diam. / mm	Pigmentation variation	Substrate / dung of:
C. albidus 1	Bos , Lepus , Felis, Canis
C. arduennensis 2	Sus scrofa
C. argenteus 3
C. aurora 5
Ascophanus aurantiacus 10	Bos
C. baeosporus 12	Cervus
C. bilobus 10	Bos
C. breviascus 10
C. breviascus	Equus
C. dextrinoideus 1	Lepus
C. dhofarensis 16	Capra
C. disculus 1	Sus
C. duplus 1
C. epithecioides	Rupicapra rupicapra
C. glaucellus 7
C. granuliformis 19
C. lacteus 20
C. leucopocillum 1
C. luteus 1
C. aff. luteus
C. marginatus 1
C. niveus 1
C. ochraceus 5
C. ochraceus	Quercus Cedrus
C. rhyparobioides	Lepus
C. sarangpurensis 17	Bos
C. sexdecimsporus 19	Sus
C. sphaerosporus 23	Equus
C. subcylindrosporus 13	Lepus
C. tetrasporus 27	Lepus Capra
C. trichosuri 24	Trichosurus vulpecula
C. uncinatus 25	Bos
C. vicinus 6	Bos
C. winteri

Table 3. Coprotus species overview - apothecial structure.

Species	Medullary excipulum	Ectoexcipular cell diam. / µm	Marginal structure	Marginal cell dim. / µm
C. albidus 1
C. arduennensis 2
C. argenteus 3
C. aurora 5
Ascophanus aurantiacus 10
C. baeosporus 12
C. bilobus 10
C. breviascus 10
C. breviascus
C. dextrinoideus 1
C. dhofarensis 16
C. disculus 1
C. duplus 1
C. epithecioides
C. glaucellus 7
C. granuliformis 19
C. lacteus 20
C. leucopocillum 1
C. luteus 1
C. aff. luteus
C. marginatus 1
C. niveus 1
C. ochraceus 5
C. ochraceus
C. rhyparobioides
C. sarangpurensis 17
C. sexdecimsporus 19
C. sphaerosporus 23
C. subcylindrosporus 13
C. tetrasporus 27
C. trichosuri 24
C. uncinatus 25
C. vicinus 6
C. winteri

Table 4. Coprotus species overview - ascus characters.

Species	Shape	Q	Dimensions / µm	Number of spores
C. albidus 1
C. arduennensis 2
C. argenteus 3
C. aurora 5
Ascophanus aurantiacus 10
C. baeosporus 12
C. bilobus 10
C. breviascus 10
C. breviascus	§	§
C. dextrinoideus 1
C. dhofarensis 16
C. disculus 1
C. duplus 1
C. epithecioides
C. glaucellus 7
C. granuliformis 19
C. lacteus 20
C. leucopocillum 1
C. luteus 1
C. aff. luteus
C. marginatus 1
C. niveus 1
C. ochraceus 5
C. ochraceus
C. rhyparobioides
C. sarangpurensis 17
C. sexdecimsporus 19
C. sphaerosporus 23
C. subcylindrosporus 13
C. tetrasporus 27
C. trichosuri 24
C. uncinatus 25
C. vicinus 6
C. winteri

Table 5. Coprotus species overview - ascospore characters.

Species	Symmetry	Shape	Poles	Dimensions / µm	Q
C. albidus 1
C. arduennensis 2
C. argenteus 3
C. aurora 5
Ascophanus aurantiacus 10
C. baeosporus 12
C. bilobus 10
C. breviascus 10
C. breviascus
C. dextrinoideus 1
C. dhofarensis 16
C. disculus 1
C. duplus 1
C. epithecioides
C. glaucellus 7
C. granuliformis 19
C. lacteus 20
C. leucopocillum 1
C. luteus 1
C. aff. luteus
C. marginatus 1
C. niveus 1
C. ochraceus 5
C. ochraceus
C. rhyparobioides
C. sarangpurensis 17
C. sexdecimsporus 19
C. sphaerosporus 23
C. subcylindrosporus 13
C. tetrasporus 27
C. trichosuri 24
C. uncinatus 25
C. vicinus 6
C. winteri

Table 6. Coprotus species overview - paraphysis characters.

Species	Apices	Width / µm	Branching	Bending	Refractive globules	Pigments
C. albidus 1
C. arduennensis 2
C. argenteus 3
C. aurora 5
Ascophanus aurantiacus 10
C. baeosporus 12
C. bilobus 10
C. breviascus 10
C. breviascus
C. dextrinoideus 1
C. dhofarensis 16
C. disculus 1
C. duplus 1
C. epithecioides	*
C. glaucellus 7
C. granuliformis 19
C. lacteus 20
C. leucopocillum 1
C. luteus 1
C. aff. luteus
C. marginatus 1
C. niveus 1
C. ochraceus 5
C. ochraceus
C. rhyparobioides
C. sarangpurensis 17
C. sexdecimsporus 19
C. sphaerosporus 23
C. subcylindrosporus 13
C. tetrasporus 27
C. trichosuri 24
C. uncinatus 25
C. vicinus 6
C. winteri