Bull. Natl. Mus. Nat. Sci., Ser. B, 37(4), pp. 147–154, December 22, 2011

Enumeration of Remarkable Japanese Discomycetes (5): First Records of
one Operculate and two Inoperculate Discomycetes in Japan
Tsuyoshi Hosoya1,*, Jae-Gu Han2, Yukiko Saito1, Zhao Yan-jie3,
Makoto Kakishima3 and Hyeon-Dong Shin2
1

Department of Botany, National Museum of Nature and Science,
4–1–1 Amakubo, Tsukuba, Ibaraki 305–0005, Japan2
Department of Environmental Science & Ecological Engineering, Korea University,Anam-Dong, Seongbuk-Gu, Seoul 136–701, Korea
3
Faculty of Life and Environmental Science, Tsukuba University,
1–1–1 Tennodai, Tsukuba, Ibaraki 305–8522, Japan* E-mail: hosoya@kahaku.go.jp(Received 3 August 2011; accepted 28 September 2011)

Abstract Three remarkable discomycetes (one operculate and two inoperculate) documented forthe first time in Japan are described and illustrated: Bisporella discedens (Helotiaceae, Helotiales),Proliferodiscus alboviridis (Hyaloscyphaceae, Helotiales), and Ramsbottomia asperior (Pyronemataceae, Pezizales). Sequences of their ITS-5.8S region (barcoding region) were obtained and
BLAST searched to confirm and evaluate their taxonomic position.
Key words : Bisporella discedens, mycobiota, Proliferodiscus alboviridis, Ramsbottomia asperior,taxonomy.

Introduction
This is the fifth part of the series on remarkable Japanese discomycetes following Hosoya etal. (2010a). Two inoperculate and one operculate
discomycetes with minute apothecia, first reported from Japan are described and illustrated.
Materials and Methods
Collection and observation procedures follow
Hosoya and Otani (1997) and Hosoya (2004).
Color codes follow the Pantone color code adopting CMYK system referring to a Pantone color
bridge (Anonymous, 2005). For previously
known distribution, the database of Global Biodiversity Information Facility (GBIF, http://data.
gbif.org/welcome.htm) was searched, and countries with occurrence of the given species areshown with an asterisk (*). Distributions known
only in literature are shown with double asterisks(**). Those with both information are shownwith triple asterisks (***). Isolates obtained fromsingle spores were used to obtain DNA from
Proliferodiscus alboviridis and Bisporella discedens. A single apothecium was used for DNA extraction for Ramsbottmia asperior. DNA extraction, PCR, and direct sequence procedure followed Hosoya et al. (2010b). The obtained DNA
sequences were BLAST searched (http://blast.
ncbi.nlm.nih.gov/Blast.cgi?PROGRAMblastn&
BLAST_PROGRAMSmegaBlast&PAGE_TYP
EBlastSearch&SHOW_DEFAULTSon&LINK_LOCblasthome), to evaluate their taxonomic
and phylogenetic affinities. The obtained extracted DNA samples are deposited in the Center for
Molecular Biodiversity Research, National Museum of Nature and Science, Japan, and available
for collaborative research.

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Fig. 1. Bisprella discedens (TNS-F-17895). A. Fresh apothecia. B. Close up of the fresh apothecia occurring ina group. Note brown hair-like structures on the margin are conidiophores. C. Section of an apothecium
showing the hymenium, ectal and medullary excipulum. D. Close up of ectal excipulum at the margin. E.
Ascus, F. Paraphyses. G. Conidiophores occurring on the ectal excipulum. H. Ascospores. C-E, G, mounted
in lactic acid. F, H, mounted in cotton blue/lactic acid. Scales. A, 1 mm; B, 0.5 mm; C, D, 50 m m; E–H,
10 m m.

Descriptions
Bisporella discedens (P.Karst.) S.E.Carp.
[Figs. 1, 2]
Basionym: Helotium discedens P.Karst., Hedwigia 28: 191. 1889.
See Carpenter (1975) for synonyms.

1.

Apothecia solitary to gregarious, patellate,shallow cup shaped or irregular due to crowded
occurrence, disc up to 0.3 mm in diameter, whitewhen fresh, pale yellow (Pantone 600PCC1 M0
Y34 K0) when dried; margin distinct, elevated andsurrounding the disc, lachnose, white with brown
particles found to be phialides in higher magnification, shallow concave, white when dried. Ectal
excipulum textura intricata, composed of thickwalled interwoven hyphae agglutinated to eachother to make each hyphal element indistinguishable, leaving 1–2 m m wide lumen, mostly arranged
parallel to, but occasionally at a low angle to themargin. The outermost cells becoming protru-

sions, short cylindrical with rounded apex or
clavate, thin-walled, occasionally differentiatedinto brown conidiophores. Conidiophores short,
simple or in groups, reduced to one to two cubic
cells, brown, bearing concolorous, Chalara-likephialides at the apex; phialides 3–4 m m at thebase, 2 m m at the apex, producing hyaline, cubic
conidia. Asci 46–564.5–5 m m, eight-spored,
cylindrical to clavate, arising from indistinct
croziers; apex conical, pore MLZ–, with or without KOH pretreatment. Ascospores 8.5–92.5–
3 m m, ellipsoid or cuneate, hyaline, often containing several oil globules. Paraphyses filiform,
1.5–2 m m thick, simple or branched at the base.
Specimens examined. HONSHU: TNS-F17895, 17897, Kikuchi Canyon, Kikuchi-shi, Kumamoto Pref. (33°028.2N, 130°5626.3E, alt.
612 m), on unidentified decaying wood. 10-X2005 col. T. Hosoya (culture FC-1228 from TNSF-17895); TNS-F-37025, Oyabu Keikoku, Hijikawa-cho, Ozu-shi, Ehime Pref. (33°2715.56N,
132°4026.55E, alt. 45 m), on unidentified

Enumeration of Remarkable Japanese Discomycetes (5)149

Fig. 2. Camera lucida illustration of Bisprella discedens (TNS-F-17895). A. Vertical section of an apothecium
through the margin showing the ectal and medullary excipulum, and the conidia producing structures on theectal excipulum. B. Ascospores. C. Conidia producing structures occurring in a group on the ectal excipulum. D. Asci. E. Paraphyses. F. Structural outline of the apothecium. Areas where short protrusions occur are
delimited by dotted line. Line drawings prepared in lactic acid mount. Scales. A–E, 10 m m; F, 100 m m.

decaying wood. 12-X-2010. Col. T. Hosoya (Culture FC-2660).
Previously known distribution. Brazil***, Dominica**, Guadeloupe**, Haiti**, Mexico*,
New Zealand***, Philippines**, Puerto Rico**,Switzerland**, Venezuela**.
Japanese name: Tsutsu-ke-shiro-byotake
Notes. This fungus is quite distinctive forhaving a Chalara (Corda) Rabenh.-like structure
on its ectal excipulum. The anamorph was firstreported as Cystodendron Bubák (Carpenter,
1975), but later identified as Bloxamia Berk. &Broome (Johnston, 1988). The ectal excipulum
composed of agglutinated thick-walled hyphaewith glassy appearance is congruent with thatof Bisporella Sacc. However, the senior author
has never obtained an anamorph from other
Bisporella isolates in Japan. As a result of

BLAST search of ITS-5.8S sequence from FC1228, HQ630988 Chalara sp. TMS-2011 voucher MS3p_50-44 was found as the best matchedsequence (Identities 530/575, 92%). The similarity of the ITS-5.8S sequence obtained from FC1228 to known sequence of Bisporella identified
to specific rank (Biporella citrina, EU940164,
AY789386,
AF335454,
AY833058,
and
GQ411507) was less than 90%. Although Bisporella is a large genus containing 19 species(Kirk et al., 2008), only two species have been
known in Japan (Katumoto, 2010). Apparently
information is not sufficient to discuss the taxonomic affinity of B. discedens. The anamorphic
state, Bloxamia was not previously documented
in Japan.
This is the first documentation of the present
fungus in Japan. The present fungus is classified

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Fig. 3. Proliferodiscus alboviridis (TNS-F-17436). A. Fresh apothecia occurring on the substrate. Note proliferation of the daughter disc (at present only sterile elements are seen) from the disc (arrow). B. Dried apothecia. C. Section of an apothecium showing the hymenium, ectal and medullary excipulum. D. Close up of asection of an apothecium at the margin showing the thick-walled ectal excipulum E. Hairs. F. Young ascus. G.
Paraphyses. H. Ascospores. C, D, mounted in lactic acid, E–H, mounted in cotton blue/lactic acid. Scales. A,
B, 1 mm; C, 20 m m; D–H, 10 m m.

in Helotiaceae, Helotiales.
2.

Proliferodiscus alboviridis (Sacc.) Spooner[Figs. 3, 4]

Basionym: Trichopeziza albovirids Sacc., Syll.
Fung. 8: 415 (1889) (ut “(Cooke) Sacc.”)See Spooner (1987) for synonyms.
Apothecia solitary to gregarious, sessile,patellate, wedged to the substrate, infrequently
proliferating from the disc; disc up to 0.5 mm indiameter, dark yellow (Pantone 123PCC0 M21
Y88 K0) rimmed with a distinct, grayish-green
hairy margin (Pantone 7542PCC24 M4 Y8
K13) when fresh, concave to urceolate, obscured
by elevated margin when dried. Ectal excipulum
textura intricata, composed of interwoven hyphae
1.5–2 m m wide, arranged parallel to the margin,
becoming thicker-walled and entangled closely,

agglutinated to form textura porrecta appearance
toward the base of the apothecium. Subicularhyphae densely interwoven, 1–2 m m wide,smooth or partially granulate, hyaline to pale.
Hairs simple, cylindrical, slightly enlarged
toward the apex, strongly granulate by having
granules of 1 m m in diameter, 3 m m wide near thebase, up to 5 m m at the tip, 30–65 m m long, 1-3septate, greyish green in mass; granules resistant
to 10% KOH. Asci 36–453–4 m m, eight-spored,
cylindrical clavate, arising from simple septa;
apex conical, pore minute, faintly MLZ without KOH pretreatment. Ascospores 4.5–61–1.5
m m, elongate ellipsoid to cuneiform, hyaline,
smooth, eguttulate, irregularly biseriate in theasci. Paraphyses cylindrical to narrow lanceolate, 1.5–2 mm thick, simple or branched at thebase. No anamorph was observed in culture.
Specimens examined. HONSHU: TNS-F-

Enumeration of Remarkable Japanese Discomycetes (5)151

Fig. 4. Camera lucida illustration of Proliferodiscus alboviridis (TNS-F-17436). A. Ascospores. B. Ascal apex
showing the reaction to Melzer’s reagent. C. Paraphyses. D. Ascus. E. Section of an apothecium at the margin
showing the ectal excipular structure. F. Structural outline of the apothecium. Line drawings prepared inlactic acid mount except for B mounted in Melzer’s reagent. Scales. A–E, 10 m m; F, 100 m m.

17436, Riku-mura, Agazuma-gun, Gunma Pref.
(36°3856.1N, 138°3833.1E), on unidentified
decaying wood. 3-VII-2006. col. T. Hosoya (culture FC-1413); TNS-F-39240, Tsukuba Botanical Garden, Amakubo, Tsukuba, Ibaraki Pref.
(38°1523.0N, 138°4514.21E), on unidentified
decaying wood (probably Prunus). 29-IV-2011.
col. T. Hosoya (culture FC-2681).
Previously known distribution. Australia**(Questionable according to Spooner, 1987),

USA**. No record in GBIF.
Japanese
name:Futasu-mi-shiro-hinanochawantake
Notes. The genus Proliferodiscus J.H.Haines
and Dumont was erected by Haines and Dumont(1983) in Polydesmiae, Arachnopezizoideae,
Hyaloscyphaceae, because it had “crisped or cork
screw-shaped” hairs and the presence of indeterminate, branched, subiculum-like hyphae at thebase of their apothecia (Haines and Dumont,

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Tsuyoshi Hosoya et al.

1983). As indicated by Haines and Dumont(1983), the irregularly curved subiculum-likehyphae are not true subicular hyphae that fix theapothecia to the substrate as in Arachnopeziza
Fuckel. Both Haines and Dumont (1983) andSpooner (1987) indicated the similarity in
apothecial structure between Lachnellula P.
Karst. and Proliferodiscus. However, Cantrell and
Hanlin (1997) showed the two genera are distantly related. The BLAST search of ITS-5.8S
sequence obtained from FC-1413 showed the highsimilarity to the registered sequence (U57990, P.
alboviridis, Identities535/545(98%)). This is
the first documentation of the genus Proliferodiscus in Japan. The present fungus is classified in
Hyaloscyphaceae, Helotiales.

3. Ramsbottomia asperior (Nyl.) Benkert & T.
Schumach.
[Figs. 5, 6]
Basionym: Peiziza asperior Nyl., Not. Sällsk.
Fauna Fl. Fenn., Förh. 10: 21. 1869.
See Benkert and Schmacher (1985) for synonyms.
Apothecia scattered to gregarious, patellate toconvex, 1.5–2 mm in diameter, orange (Pantone
1495CR255 G145 B51) when fresh, becoming
discoid to shallow cupulate, brownish yellow(Pantone 135PCC0 M21 Y74 K0). Ectal exciplum textura globulosa composed of globular torectangular cells 30–5020–40 m m, outermost
cells protruding into hyphoid hairs up to 75 m mlong, 10–15 m m wide. Ascospores globose, 13–

Fig. 5. Ramsbottomia asperior (TNS-F-36001). A–C. Fresh apothecia on ground. D. Vertical section of anapothecium at the margin showing the ectal excipulum. E. Asci. F. Ascospores. G. The same ascospores in F
at different focus. Note spines on the ascospore wall. H–I. Ascospores in cotton blue/lactic acid. Note spines
are stained by cotton blue. J. Young ascospores showing the globular cell content. K. Paraphyses in MLZ,
showing the cellular contents stained. Scales. A, B, 1 mm, C, 2 mm, D, 50 m m; E, 20 m m. F-K, 10 m m.

Enumeration of Remarkable Japanese Discomycetes (5)153

Fig. 6. Camera lucida illustration of Ramsbottomia asperior (TNS-F-36001). A. Vertical section of an apothecium at the margin showing the ectal and medullary excipulum and an ascus. B. Paraphyses. One at the rightshows the cell contents stained in MLZ. C. Mature ascospore. D. Young ascospores showing globular cell
contents. E. Structural outline of the apothecium. Line drawings prepared in lactic acid mount except for one
at the right in photo B. Scales. A, 50 m m; B–D, 10 m m; E, 1 mm.

15 m m in diameter excluding the spines; smooth,spumose to multi-guttulate, stained brown in
MLZ when young, becoming spiny, having remarkable, conical spines up to 3 m m long, stained
blue in cotton blue in lactic acid; eguttulate whenmature, uniseriate in the asci. Asci ca. 30018–20 m m, eight-spored, thin-walled, MLZ—with orwithout KOH pretreatment, arising from croziers.
Paraphyses straight, septate, occasionally slightly enlarged at the apex, up to 4–5 m m thick; apical cells 5–7.5(–10) long, having carotenoid
granules stained blue in MLZ.
Specimens examined. HONSHU: TNS-F-

36001, Tsukuba Botanical Garden, Amakubo,
Tsukuba, Ibaraki Pref. (38°1523.0N, 138°45
14.21E), on ground. 27-VI-2010. col. Y. Kondo.
Previously known distribution. Austria*,
Brazil*, Finland**, Germany***, Iceland*, Morocco*, New Zealand*, Mexico*, Norway*,
Scottland**, Sweden***, Switzerland*, UK*,
USA***.
Japanese name: Togemino-daidai-saratake
Notes. The genus Ramsbottomia Buckley
was erected by Buckley (1923) based on a single
species R. lamprosporoidea. The generic concept
was verified by Kullman and Brummelen (1992)

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Tsuyoshi Hosoya et al.

who justified its segregation from Lamprospora
De Not., a genus with globose spores andbryophilous habitat. Yao and Spooner (1995)
claimed the delimitation of Ramsbottomia andLamprospora requires further investigation, andRifai (1968) listed Ramsbottomia as a synonymof Lamprospora. However, Perry et al. (2007)
concluded that Ramsbottomia can be segregatedfrom Lamprospora as well as Octospora Hedw.
and Neottiella (Cooke) Sacc. based on molecularphylogeny.
Currently, three more species are accepted in
Ramsbottomia: R. lamprosporoidea W.D.Buckley, R. crec’hqueraultii (P.Crouan & H.Crouan)
Benkert & T.Schumach. and R. macrantha(Boud.) Benkert & T.Schumach. (Kirk et al.,2008). Benkert and Schmacher (1985) who
emended the genus regarded R. lamprosporoidea
synonymous to R. asperior. However, Yao andSpooner (1995) claimed it is distinguished byhaving longer spines (2–6 m m while the spines in
R. asperior are up to 3.5 m m) on the ascosporesand presence of colored hairs. The characteristicsof the present specimen almost agree with thedescription given for R. asperior in Benkert andSchmacher (1985) except for the ascospores in
the present specimen are almost globose whilethat in R. asperior are elliptic to globose.
Obtaining a culture was unsuccessful because
the spores did not germinate.
This is the first documentation of the genus
Ramsbottomia in Japan. The present fungus isclassified in Pyronemataceae, Pezizales.
Acknowledgments
The author wish to thank Dr. R. P. Korf, Prof.
Emeritus, Cornell University for kindly reviewing the manuscript.
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