Ascomycete.org 5

Wolfina aurantiopsis, a rare species in the family
Chorioactidaceae (Pezizales)
Carlo AGNELLO
Matteo CARBONE
Christine BRAATEN
Ascomycete.org, 5 (1) : 39-45.
Janvier 2013
Mise en ligne le 03/01/2013
Summary: A detailed morphological study of the rare ascomycete Wolfina aurantiopsis (Ellis) Eckblad is presented after the thorough study of two collections from Tennessee (USA). Macroscopic and microscopic
color photographs as well as line drawings are included. Notes regarding taxonomy and nomenclature of
W. aurantiopsis and related species are proposed.
Keywords: Ascomycota, Galiella, Trichaleurina, Sarcosoma, Wolfina oblongispora, Wolfina papuana, Tennessee, eastern USA, taxonomy.
Riassunto: Viene presentato uno studio morfologico dettagliato della rara Wolfina aurantiopsis (Ellis) Eckblad
basato su due raccolte effettuate nel Tennessee (USA). Lo studio è corredato da fotocolor in habitat, foto al
microscopio e disegno a tratto. Vengono inoltre fornite note tassonomico-nomenclaturali e discusso un
confronto con specie simili.
Parole chiave: Ascomycota, Galiella, Trichaleurina, Sarcosoma, Wolfina oblongispora, Wolfina papuana, Tennessee, U.S.A. orientali, tassonomia.
Résumé : une étude morphologique détaillée du rare ascomycète Wolfina aurantiopsis (Ellis) Eckblad est
présentée d’après l’examen approfondi de deux récoltes du Tennessee (États-Unis). Des photographies macroscopiques et microscopiques ainsi que des dessins au trait sont ajoutés. Des notes concernant la taxinomie et la nomenclature de W. aurantiopsis et des espèces associées sont proposées.
Mots-clés : Ascomycota, Galiella, Trichaleurina, Sarcosoma, Wolfina oblongispora, Wolfina papuana, Tennessee, est des États-Unis, taxinomie.
Introduction
Materials and methods
The genus Wolfina was established by SEAVER (1937) and was
based on Peziza aurantiopsis Ellis. ECKBLAD (1968) then validated the
genus due to the previous lack of a Latin diagnosis (required at that
time, ex art. 36.1 I.C.B.N.): “Apothecia juvenilia clausa, matura dehiscentia, cupulata, excipulo toto textura intricata constructo, extus tomentosa, tomento e stratis cellularibus externis formato; pili tomenti
semplice, septati, recti vel undulati, pariete fusco, crasso instructi, laeve
vel basin versus exasperati. Asci cylindrici, non amyloidei, octospori, pariete crasso instructi. Sporae ellipsoideae, hyalinae, non guttulatae,
leaves vel longitudinaliter leviter striatae. Paraphyses filiformes, rectae,
septatae”. Finally, the genus was placed into the family Chorioactidaceae Pfister by PFISTER et al. (2008).
Until now, only two other species have been described in this
genus, Wolfina oblongispora (J.Z. Cao) W.Y. Zhuang & Zheng Wang
from China and Wolfina papuana Otani from Papua New Guinea (see
discussion). However, further molecular and systematic study is needed to determine their phylogenetic placement.
After careful evaluation of the family Sarcosomataceae Kobayasi,
the authors of this paper were obliged to consider and explore the
sister family Chorioactidaceae Pfister (PFISTER et al., 2008). Phylogenetic studies conducted on Sarcosomataceae by CARBONE et al.
(2013), lead to the accommodation of two additional genera into
Chorioactidaceae: the new genus Pseudosarcosoma M. Carbone,
Agnello & P. Alvarado, and the established genus Trichaleurina
Rehm. Nevertheless, our knowledge of the four original Chorioactidaceae genera (i.e., Chorioactis Kupfer ex Eckblad, Desmazierella Lib.,
Neournula Paden & Tylutki and Wolfina Seaver ex Eckblad) was limited to Neournula pouchetii (Berthet & Riousset) Paden only.
Wolfina aurantiopsis has a presumably restricted geographic
range limited to an area comprising several states in the eastern United States. Recently, WANG (2011) reported this species from Taiwan,
but we believe that it should be investigated genetically to figure
out if it is conspecific with the American collections. Anyway, after
thorough examination of collections made from Fall Creek Falls
State Park and the Great Smoky Mountains National Park (GSMNP)
in Tennessee, we are now able to present a detailed study of this uncommon species.
Microscopical characters are based on fresh and dry specimens.
Three optical microscopes were used: Nikon Eclipse E600 compound microscope, Olympus CX41 trinocular and Optika B353 trinocular with plan-achromatic objectives 4×, 10×, 40×, 60× and 100×
in oil immersion. The following main reagents were used: Melzer’s
reagent, cotton blue and Congo red. Water mounts were used for
the observation of the pigmentation and spore size. At least 30
spores were measured from each apothecium and from spore deposit.
Taxonomy
Wolfina aurantiopsis (Ellis) Seaver ex Eckblad, Nytt. Mag. Bot.,
15(1-2): 126 (1968).
Basionym: Peziza aurantiopsis Ellis, Bull. Torrey Bot. Club, 9: 18
(1882).
Synonymy:
≡ Lachnea aurantiopsis (Ellis) Sacc., Syll. Fung., 8: 180 (1889); Scutellinia aurantiopsis Kuntze, Rev. Gen. Pl., 2: 869 (1891).
= Sarcosoma carolinianum Durand, J. Mycol., 9: 103 (1903), fide
SEAVER (1937).
Original diagnosis: “Peziza (Sarcoscypha) aurantiopsis – Sessile,
about one inch across, with a coarse, felt-like, black-brown mycelium at the base, matted together on hardwood branches, etc., on
which it grows; the outside of the cup also coated with coarse
(.00025’ diam.) brown, smooth sparingly-branched, continuous
hairs; disk clear pale yellow, nearly sulphur yellow, becoming dull
orange when dry; margin thin, pale, narrow, erect; flesh white, thick
(1/8’-1/4’), firm, elastic, dry; asci cylindrical, subtruncate above,
.0013-.0014 × .0006’; paraphyses stout, septate, slightly thickened
above; sporidia oblong-elliptical, granular, .0009’-.0010’ × .0006’,
epispore smooth. On the bare soil and on decaying wood and leaves
in low, sandy, oak and pine woods, Sept. 1881.”
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Description of studied collections
Macroscopical characters
Apothecium shallow cup-shaped, sessile, 2.5–4.5 cm wide. Hymenium shallow, pale yellow to pale orangish-ochraceous in age,
smooth. External surface rough, black, convoluted. Flesh thick,
white, firm, corky in dried specimen; odor and taste not distinctive.
clavate-cylindrical, narrowed below to a long, slender pedicel, apex
rounded, not blue with iodine, 400-450 × 18 μ, opening by a lid.
Spores 8, uniseriate, hyaline, smooth, continuous, elliptical, 25-30 ×
15 μ (the majority 28 × 15 μ); Paraphyses cylindrical, hyaline, septate, very little thickened at the free tips, 3 μ thick”.
Notes on similar species
Microscopical characters
Asci cylindrical, 340–430 × 14.5–20 μm, operculate, inamyloid, 8spored, with walls up to 2 μm thick; base tapering and flexuous. Paraphyses filiform, septate, anastomosing, 3–4.5 μm wide, as long as
the asci, not or only slightly enlarged at the tips up to 5 (–5.5) μm.
Spores elliptical to subcylindrical, 25.4–32 × 10.4–15 μm (on average 28.3 × 12.5 μm), Q = 2.03–2.52 (average 2.27), hyaline, with granular content, sometimes embedded in a gelatinous sheath; the
surface is marked by thin longitudinal, sometimes anastomosing
striations. Subhymenium of a thick textura intricata made up of cylindrical, septate, 5–8 (–9) μm wide hyphae, arranged perpendicular
to the asci; brownish at low magnification. Medullary excipulum
of textura intricata made up of hyaline, closely septate, 5–8 (–10) μm
wide hyphae. Ectal excipulum: not a real ectal excipulum but a simple extension of the medullary excipulum where darker hyphae are
arranged in palisade, mixed with basal part of hairs. External hairs
cylindrical, 5–9 (–12) μm wide, septate, with walls up to 1 μm thick,
brown, entirely covered by warts or prickles when young or not well
developed, then smooth in almost their length except in the basal
part where they remain ornamented.
Ecology and studied collections
Solitary or in small clusters, broadly attached to fallen twigs and
branches of hardwood in a section of a trail dominated by Rhododendron maximum.
UNITED STATES OF AMERICA. Tennessee, Van Buren County, Fall Creek
Falls State Park, 518 m a.s.l., 15 July 2012, legit and det. C. Braaten,
herbarium number TENN 67714. Tennessee, Cocke County, Great
Smoky Mountains National Park, Low Gap Trail, 610 m a.s.l., 6 August 2012, legit and det. C. Braaten, herbarium number TENN 67128,
Genbank numbers (LSU - KC306743; ITS - KC306744).
Discussion
Brief historical and nomenclatural notes
Originally described from New Jersey (U.S.A.) as Peziza aurantiopsis (ELLIS, 1882), Wolfina aurantiopsis was then transferred first in the
genus Lachnea by SACCARDO (1889) and subsequently in Scutellinia
by KUNTZE(1891).
SEAVER (1911) applied the name Lachnea aurantiopsis, however he
(SEAVER, 1937) made the decision to include this species into his new
genus Wolfina due to its apparent lack of affinity with genera that
had been described at that time. In addition, SEAVER (1937) revised
the type specimen of Sarcosoma carolinianum Durand (DURAND,
1903) and synonymized it with Wolfina aurantiopsis because no
morphological differences were observed. We have not yet examined Durand’s type material, however, given the convincing characterization from Durand’s original description, at present we do not
have reason to doubt the synonymy. The original description of Sarcosoma carolinianum is as follows:
“Plants solitary, sessile, attached by a dark brown tomentum; at
first closed, then opening by a pore at the apex, expanding and enlarging finally becoming saucer-shaped, up to 4 cm. diam. Disk
tawny-ochraceous, externally brown, covered with a thick, appressed, brown tomentum, threads very long 7-8 μ thick, septate, rather
shining, but little wrinkled; substance tough-gelatinous, not at all
watery, so that the plant nearly retains its shape, size and color when
dry. Consistency of the dry plant corky, not horny and brittle, and
exterior nearly even. Flesh white, excipulum composed entirely of
interwoven hyphae which are thick and septate, 5-6 μ diam. Asci
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Morphological characters reported here are clearly identical with
those made by many authors (DURAND, 1903, as Sarcosoma carolinianum; SEAVER, 1911, 1937, 1942; ECKBLAD, 1968; ROODY, 2003; BESSETTE et al., 2007; PFISTER et al., 2008). The only exception is in ELLIS
(1882) where we find slightly smaller spores of ca. 25 × 15 μm. Although collections of this species have been recorded from a wide
but confined area in the eastern United States (PFISTER et al., 2008),
we decided to discuss some similar species not present in this area.
Wolfina oblongispora originally described as Galiella oblongispora
J.Z. Cao (CAO et al., 1992), was then amended and transferred to Wolfina by ZHUANG & WANG (1998). The most emphasized difference is a
larger spore size, 36–45 × (15–) 17.5–22 μm, however, as reported by
ZHUANG & WANG (1998) this species is characterized by spores ornamentated by longitudinal striations mixed with some warts. The original description: “Apothecia sessilia vel breviter stipitata, obconica,
3.5-4.5 cm alta, 3-5 cm lata. Hymenium flavidum vel alboluteum. Facies exterior pilei nigro-fusca. Hypothecium distinctum, denso-intricatum. Excipuli medullaris textura intricata in matrica gelatinosa sita.
Excipuli exterior 80-100 μm crassa, ad texturam angularem vel globulosam, cellulis brunneo-tunicatis. Capillae brunneo-tunicata, flexuosae, 80-600 × 7-12.5 μm. Asci longo-cylindrici, I-, 8-spori, 400-450 ×
18-21 μm. Ascosporae uniseriatae, oblongae, 33-42 × 16-18 μm, verruculosus. Paraphyses filiformis, 2.5-3 μm diam”.
Wolfina papuana Otani (OTANI, 1975) was tentatively included in
Wolfina, with the following Latin diagnosis: “Apothecia sparsa, sessilia, disco tenuiter concavo vel fere plano margine humili, circa 1.52.0 cm in diametro, flavo ubi novo salmoneo ubi exsiccato, receptaculo
hemisphaerico carne gelatinosa crassa, circa 1 cm crasso, extus rugoso,
castaneo, tomentoso. Excipulum externum tenue 17.5-25.0 μ crassum,
strato 2- vel 3-cellularum, cellula extima brunneitunicata oblongata
5.0-10.0 × 5.0-7.5 μ, pilis longis simplici ab cellulis extremis projectis,
cylindraceis, obtusis, tenuitunicatis, laevibus, septatis, undatis, laxe
conglomeratis, brunneis circa basim, superne pallidioribus; excipulum
medullosum maxime crassum, circa 3-5 mm vel plus etiam, textura intricata , hyphis tenuibus, 2.5-3.0 μ in diametro, laxe intertextis, in gelatina inclusis; hymenium circa 800-1000 μ crassum, massis
columelliformibus crystallorum flavorum cubicorum vel oblongorum
interspersum. Asci cylindracei, longi, apud basim angustati, crassi tunicati, suboperculati, liquori jodi non caerulescentes, octospori, 600650 × 15.0-17.5 μ; ascosporae longifusiformes, non-septatae, laeves,
flavidae, modice crassitunicatae, guttulatae, 42.5-52.5 × 13.5-15.0 μ,
paraphyses anguste cylindraceae, septatae, rectae, semplice, 2.5-3.0 μ
in diametro, contentis flavidis”. We are not currently aware of any further collections or descriptions of this species and so we can only
use the protologue to summarize the following important characters: gelatinized context, fusiform spores, smooth external hairs, presence of crystals in the hymenium, thin ectal excipulum consisting
of 2–3 layers of elements. Our opinion is consistent with that of
ZHUANG & WANG (1998) in concluding that this species should be excluded from the genus Wolfina.
Macroscopically, biogeographically, and ecologically, Galiella rufa
(Schwein.) Nannf. & Korf appears akin to W. aurantiopsis. However,
G. rufa is well distinguished by globose to shallow cupulate (sub)sessile apothecia with a irregularly toothed margin, reddish-brown to
orange-brown hymenium, blackish-brown external surface covered
with hairs that give it a tomentose appearance; the flesh is gelatinous and rubbery; microscopically it is distinguished by the subfusoid ornamented spores (SEAVER, 1928, 1942; BESSETTE et al., 1997;
PHILLIPS, 2005). The genera Galiella Nannf. & Korf and Trichaleurina
Rehm are in need of revision. In any case, all species (tropical or tem-
perate) retained in these two genera possess subfusoid to fusoid,
warted spores.
Sarcosoma globosum (Schmidel: Fr.) Casp. is recorded from eastern North America (BESSETTE et al., 1997) and is easily distinguished
macroscopically by a larger and more fleshy apothecium with a dark
hymenium. Microscopical features are also particularly distinct due
to the moniliod external hairs (present in immature specimens too).
Notes on hymenial hairs
As reported in CARBONE et al. (2013), the term “hymenial hairs” —
or setae in the sense of PADEN (1983) — refers to structures that are
not true hymenial hairs, or setae, because these designations are
better applied, e.g. to the hymenial elements of the genus Tricho-
glossum Boud. However, from the time of their discovery in Sarcosomataceae, the term “hymenial hairs” has been used for those elements distinguished from the true paraphyses by the lack of septa
and a larger width. It must be noted that sometimes the hymenial
hairs are not easily detected, in fact, in some species they can be
disguised among bundles of paraphyses cohered by an amorphous
exudate.
The absence of hymenial hairs in W. aurantiopsis is noteworthy
due to our observation that other species in the family Chorioactidaceae (i.e. Neournula pouchetii, Pseudosarcosoma latahense, Trichaleurina javanica and T. celebica) also lack this microscopic
character. Moreover we currently can say that this feature is present
in all studied Sarcosomataceae species.
Plate 1 – Wolfina aurantiopsis. Pictures: C. Braaten.
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Plate 2 – Wolfina aurantiopsis. Microscopic characters.
A: Spores in ascus in Congo red. B: Empty ascus with operculum in Congo red. C: Upper part of paraphyses in Congo red.
D-N: Spores in water. Scale bars: all = 10 μm except C = 5 μm.
Pictures: C. Agnello & M. Carbone.
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Plate 3 – Wolfina aurantiopsis.
A: Section of a fresh apothecium, with highlighted parts used for pictures B and E.
B: Section. C: Medullary excipulum. D: External part of the excipulum. E: External part of the excipulum and external hairs.
F: Young external warted hairs. G: Smooth and warty external hairs. H: Young warty external hairs.
I: External hair with tip becoming smooth with growth. Scale bars: all = 10 μm except E = 50 μm.
Pictures: C. Agnello & M. Carbone.
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Plate 4 – Wolfina aurantiopsis. Microscopic characters.
A: Section. B: Spores. C: Ascus tip. D: Paraphyses. E: External hairs and lower part of the excipulum.
Drawing: C. Agnello.
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Acknowledgements
We want to thank Dr. Pablo Alvarado (Alvalab) for his service of
DNA sequencing and for the assistance in interpreting the results.
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Carlo Agnello
Via Antonio Gramsci 11
72023 Mesagne
Italy
[email protected]
Matteo Carbone
Via Don Luigi Sturzo 173
16148 Genova
Italy
[email protected]
Christine Braaten
University of Tennessee, Knoxville
TN 37996-1610
USA
[email protected]
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