Hamatocanthoscypha rotundispora Raitv. & R. Galán

Kušan, Ivana, Matočec, Neven, Antonić, Oleg & Hairaud, Michel, 2014, Biogeographical variability and re-description of an imperfectly known species Hamatocanthoscypha rotundispora (Helotiales, Hyaloscyphaceae), Phytotaxa 170 (1), pp. 1-12 : 2-7

Hamatocanthoscypha rotundispora Raitv. & R. Galán in Galán & Raitviir (1994: 459)

Fig. 1–3

Description:

Apothecia superficial, solitary, gregarious, cup-shaped, stipitate. When fresh, disc 0.1–0.8 mm {3}, in statu sicco up to 0.3 mm {2} in diameter, concave to plane, white (1A1), stipe cylindrical, 0.1–0.3 mm {3} high, up to 0.1 mm {2} wide, base greyish yellow (4B4). Hymenium white (1A1), smooth, unwrinkled, normally not translucent. Margin sharp, slightly elevated above hymenium, with dense white (1A1) shortly protruding hairs, ground white (1A1). Excipular and stipe surface concolourous; very sparsely beset with shorter hairs. Subiculum, associated asexual states or stromatic tissues not observed.

Hymenial elements: Hymenium * 38–45 µm {2}, † 33–42 µm {2} thick. Asci * 30–43.2 × 4.9–5.8 µm {3}, † 26–39.5 × 3.7–5.4 µm {4}, * asci protruding above paraphyses for 4.8–9.1 µm {1}, pars sporifera * 10–14.3(–20.2) µm {3}, 8- spored, spore arrangement biseriate in living but uniseriate in dead asci, cylindric-subventricose, with conical-truncate apex and ± truncate base, apex seemingly without apical ring and barely thickened in * asci, but considerably thickened and with faintly visible apical ring in † IKI, arising from croziers {4}, often 2–4 asci arise from a single crozier cell, in */† IKI completely inamyloid, in * CRB walls not stained. Ascospores * 3.3–5.2(–5.7) × (1.9–)2–3(–3.2) µm {3} (n = 150), † (2.2–)2.4–4(–4.2) × (1.1–)1.4–2.3(–2.6) µm {4} (n = 200), descriptive statistics of ascospore morphometrical variables (L, W, Q and V) is given in Table 1.; subciborioid to ciborioid to rarely amygdaliform in lateral (side) view and limoniform to oculiform in dorsiventral (front) view, also fusiform and piscioid in dead spores, bilateral symmetric and slightly heteropolar, 1-celled, wall hyaline and strongly refractive, 0.2–0.3 µm {1} thick, smooth; containing 1–3(–5) LBs that are positioned eccentric or near poles, * 0.2–0.8 µm {2} in diam., rarely eguttulate, after ejection spores are aggregated together and each enveloped by a delicate sheath; in * CRB with one to a few globose greyish violet (19C7) MCs, * 0.3–2 µm {3} in diam. that are dissolving in † CRB-KOH; in * CRB sporoplasm and walls not stained, in * IKI sporoplasm light yellow (3A5) to yellow (2A7), without glycogen accumulations. Paraphyses cylindric-obtuse to subclavate, sparse, simple, not branching, hyaline, sometimes with thin, hyaline, moderately refractive exudate in apical part, apical cells * 15–35 × 2–3.2 µm {3}, † 1.4–2.1 µm {3} wide, each apical cell with a single cylindrical, hyaline, very slightly refractive VB which is either completely absent in dead cells or present as irregular remnants; in * CRB VBs turquoise (24A8), but instantly vanish in † CRB-KOH; in * IKI cytoplasm not notably stained. Subhymenium scarcely distinguishable from medulla, composed of hyaline, thin-walled hyphae and ascogenous cells, layer * 7–11.8 µm {2}, † 4–6 µm {2} thick.

Margin: submarginal tissue composed of hyaline textura prismatica, cells ± parallel to surface, * 7.6–13.2 × 2–2.6 µm {1}, thin-walled, not gelatinised, outer cells of margin abundantly beset with hairs, these strongly curved, uncinate to helical, partly intertwined, with outwardly bent tips, tapering toward apex but not pointed, 2–3(–4)-celled, * 28–55 × 2–4.1 µm {2}, at apex * 0.9–1.6 µm {2} wide, † (20–)37–56(–62) × 2.9–4.1 µm {4}, at apex † 0.9–1.3(–1.5) µm {4} wide, base sometimes swollen at lower positioned hairs, basal cells occasionally with hyaline and moderately refractive RCGs, apical cell wall rather sparsely covered with flat to granular, hyaline, highly refractive exudate; exudate in * CRB turquoise (24A8), in † CRB-KOH stable and becoming light lilac (15A5), RCGs in * CRB greyish violet (19C7), disappearing in † CRB-KOH; in */† IKI exudate pastel yellow to light yellow (3A4-3A5) or rarely reddish orange (7B8) and cytoplasm granular yellow (2A6); in * IKI RCGs highly contrasted, in MLZ exudate not stained and partly dissolved.

Excipular flank: Medullar excipulum reduced, hyaline, layer * 6.2–9 µm {1}, † 5–8(–12) µm {2} thick, composed of sparse, narrow, cylindrical hyphae running parallel to receptacle surface (textura porrecta-prismatica), at apothecial centre textura porrecta-intricata, hyphae * (0.9–) 1.4–2.7 µm {2}, † 1.1–2.2 µm {2} wide, with thin and not gelatinised walls; * IKI-, * CRB-, MLZ-. Ectal excipulum composed of hyaline textura prismatica with cells oriented parallel to the surface, layer * 8.2–12.5 µm {2}, † 6–10 µm {2} thick at middle flank, cells * 5.3–14.1 × 2.4–6.5 µm {2}, † 5.3–16.5 × 1.7–6 µm {3}, ±thin-walled, not gelatinised, some outermost cells contain sparse LBs but more often RCGs that are hyaline and of moderate to high refractivity; in * CRB cytoplasm greyish blue (20B4), cell walls not stained, RCGs greyish violet (19C7) and instantly vanishing in † CRB-KOH, LBs stable; in * IKI RCGs and LBs not stained. Outermost cells give rise to hyaline hairs from upper flank to the stipe, 1(–2)-celled, * 5–26 × 2.2–3.7 µm {2}, at apices * 1.1–2.5 µm {2} wide, † 5.1–23.4(–33) × 2–4.1(–5) µm {4}, at apices † 0.9–2 µm {4} wide, thin-walled, smooth or faintly encrusted, bent outwards, apex rounded to slightly tapered, not pointed, shorter and less bent than marginal hairs, forming pustulate aggregations (2–5 hairs per aggregation); * IKI-, * CRB-, MLZ-.

Stipe composed of a medullar (central) and an ectal layer and of a histologically differentiated basal part. Medullar part hyaline, thin-walled textura porrecta, hyphae ±vertically oriented, † 1.5–2.1 µm {1} wide. Ectal layer hyaline, thinwalled textura prismatica, † 6–9 µm {1} thick, cells oriented at a very high angle to the stipe surface, † 1.8–3.5 µm {1} wide, with sparse RCGs; outermost cells giving rise to hyaline hairs, 1(–2)-celled, † 7.1–22.6 × 2–3.5 µm {1}, at apices † 1.6–2.1 µm {1} wide, cylindric-obtuse to very slightly tapered, apices curved to uncinate and bent downwards, with very sparse, granular, hyaline, moderately refractive exudate on their surface. Basal part of densely packed, thick-walled, pigmented textura prismatica -angularis, this part * 40–65 × 70–80 µm {1}, cells ±vertically oriented, prismatic cells * 6–8.8 × 2–3.5 µm {1}, more isodiametric cells * 4.2–5.1 µm {1} wide, walls glassy, light yellow (2A5), highly refractive, not gelatinised, * cytoplasm subhyaline; surface of basal part covered with scanty greyish yellow (4B4) cylindrical basal hyphae, individual cells † 5–10.1 × 2.3–4.8 µm {1}, moderately thick-walled, wall † 0.3–0.6 µm {1} thick.

Specimens examined:

— CROATIA. Dalmatia: Island of Korčula, SE from Lumbarda, Pržina cove, 8 m asl, 42°55’06’’ N, 17°11’13’’ E, habitat: degraded thermo-mediterranean fully evergreen maquis on sandy soil with Juniperus phoenicea , Pistacia lentiscus , Olea sylvestris , Smilax aspera , Arundo donax , Agave americana and Olea europaea , substrate: rotten twigs and leaves of Juniperus phoenicea in litter, associated with Pithya cupressina , 31 December 2010, M. Vojinović (CNF-2/8847!).

— FRANCE. Poitou-Charentes: Charente-Maritime, Island of Ré, Forêt Domaniale de la Combe à l’Eau (Arsen-Ré), 7 m asl, 46°12’55’’ N, 1°32’14’’ E, habitat: 25 years old plantation of Cupressus macrocarpa on sandy ground, substrate: C. macrocarpa leaves and branchlets still attached to cut branches lying on ground, 2 collections: (1) associated with Chloroscypha alutipes and Boubovia ascoboloides , 10 December 2011, M. Hairaud ( M. H. 071211), duplex (CNF- 2/9110!) and (2) associated with Cistella acuum , Chloroscypha alutipes and B. ascoboloides , 31 December 2011, M. Hairaud ( M. H. 231111), duplex (CNF-2/9113!).

— FRANCE. Bretagne: Côtes d’Armor, Tourony (Trégastel), 4 m asl, 48°49’34’’ N, 3°29’36’’ E, habitat: public park, substrate: branches of C. macrocarpa lying on soil, 13 November 2009, B. Capoen ( B. C. 141109), duplex (CNF- 2/8889!) .

— SPAIN. Guadalajara : Tamajón , Ermita de los Enebrales, on fallen decaying leaves of Juniperus thurifera , 3 December 1991, R. Galán & A. Raitviir ( AH 6696 ! = R. G. 6696, isotype, designated by Galán & Raitviir 1994) .

Notes: The protologue of H. rotundispora ( Galán & Raitviir 1994) designates TAA-136197 as holotype and R. G. 6696 [= AH 6696] as isotype, although on the exsiccata label that we received from AH, R. G. 6696 is clearly indicated as holotype, with an isotype deposited in Ain Raitviir’s herbarium ( A. R., in TAA). We were informed by Ilmi Parmasto (pers. comm.) that the specimen TAA-136197 is inaccessible/misplaced. Therefore we here rely on the study of the isotype material which consists of about 30 mature apothecia and also includes a slide containing a completely dried section.

— SPAIN. Navarra, Rada, Laguna de Rada , 365 m asl, 42º18’09’’ N, 1º34’52’ W, substrate: on rotten leaves of Cupressus sp. , 6 January 2013, M. Tapia (ERD-5751) .

FIGURE. Hamatocanthoscypha rotundispora (CNF-2/8847). a. * Asci with a part of ascogenous system and paraphysis in H 2 O. b. * Ascus tip in H 2 O. c. * Ascospores in H 2 O, some with LBs (transparent circle). d. * Ascospore in CRB, with LB (transparent circle) and MCs (shaded circles). e. * Apothecial tissues and hairs at margin, in H 2 O, with LB (transparent circle) and RCGs (shaded circles). ↔ display dorsiventral (front) spore view, → display lateral (side) spore view (arrow directed towards dorsal side). Del. N. Matočec.

FIGURE. Hamatocanthoscypha rotundispora —apothecia and hymenial elements. a–b. Living apothecia. c–e. * Asci in H 2 O. f. * Asci and paraphysis in H 2 O. g. † Asci in KOH. h. * Croziers in H 2 O. i. * Asci in IKI. j. † Asci in IKI. k. * Asci in CRB. l–o. * Ascospores in H 2 O. p. † Ascospores in H 2 O. r. * Ascospores in CRB. g and p from AH 6696 (holotype), a, c, e, h, j, k, o and r from CNF-2/8847, b from CNF- 2/9110, d, f, i, l, m and n from CNF-2/9113. Scale bars: g, h, j–r= 5 µm, d, i= 10 µm, c, e, f= 20 µm, a= 0.5 mm, b= 1 mm. All photos N.

Matočec & I. Kušan except b which is taken by M. Hairaud.

FIGURE. Hamatocanthoscypha rotundispora —sterile apothecial tissue. a. * Apothecial tissues and hairs in H 2 O. b. * Marginal hairs in H 2 O. c. † Marginal hairs in KOH. d. * Marginal hairs in IKI. e. * Marginal hairs in CRB. f. * Surface of excipular flank in H 2 O. g. * Excipular flank with hymenium in H 2 O. h. * Squashed ectal excipulum in CRB. i. † Ectal excipulum in CRB-KOH. j. * Stipe basal part in H 2 O. k. * Stipe surface in H 2 O. Arrows indicate RCGs except in i where it shows LB. c from AH 6696 (holotype), a, g and k from CNF-2/8847, b, d–f and h–j from CNF-2/9113. Scale bars: b, e, f, h–k= 10 µm, a, c, d, g= 20 µm. Phot. N. Matočec & I. Kušan.

Descriptive statistics for each of four morphometrical variables (L, W, Q and V) and for each of six statistical samples are given in Table 1. Results of Shapiro-Wilk test suggest that significant deviation of normal distribution exists in all morphometrical variables and also in all statistical samples (although not in the each variable-sample combination). Consequently, it can be concluded that non-parametric statistics (median, quartiles) is more appropriate for description of morphometrical variables distributions than parametric statistics. According to this, a non-parametric Mann-Whitney test was used for statistical sample comparison (see Table 2).

TABLE. Descriptive statistics of Hamatocanthoscypha rotundispora ascospore morphometric variables (N = 50 for each of six statistical samples). StD—standard deviation; S-W and p(S-W)—Shapiro-Wilk statistics for normality and respective probability (bold denotes statistical significance at probability level of p = 0.05); Min, L_Quart, U_Quart and Max—

minimum, lower quartile, upper quartile and maximum.