Bull. Natl. Mus. Nat. Sci., Ser. B, 45(2), pp. 47–55, May 22, 2019

Enumeration of Remarkable Japanese Discomycetes (11):Notes on Three Species of Lachnaceae New to Japan
with One New Combination
Yukito Tochihara1, Rei Sasagawa2 and Tsuyoshi Hosoya3*
1
Department of Biological Sciences, Graduate School of Science,
The University of Tokyo, 7–3–1 Hongo, Bunkyo-ku, Tokyo 113–0033, Japan
2
Faculty of Life and Environmental Science, Tsukuba University,
1–1–1 Tennodai, Tsukuba, Ibaraki 305–0005, Japan3
Department of Botany, National Museum of Nature and Science,
4–1–1 Amakubo, Tsukuba, Ibaraki 305–0005, Japan* E-mail: hosoya@kahaku.go.jp(Received 12 February 2019; accepted 27 March 2019)

Abstract Three species of Lachnaceae new to Japan are described and illustrated with one new
combination: Brunnipila clandestina, B. pseudocannabina (new combination), and Incrucipulum
capitatum.
Key words : Brunnipila clandestina, Brunnipila pseudocannabina, Incrucipulum capitatum,
mycobiota, taxonomy.

Introduction
This is the eleventh part of the series of remarkable Japanese discomycetes following Hosoya andZhao (2016) to elucidatee Japanese cup-fungi
mycobiota. Three species of Lachnaceae Raitv. are
newly documented from Japan, including one new
combination. Descriptions, micrographs and line
drawings of three species are provided.
Materials and Methods
Collection, isolation and observation procedures
mainly followed Hosoya and Otani (1997) andHosoya (2004). Examination of micromorphology
was carried out using cotton blue dissolved in water(CB/W) or cotton blue dissolved in lactic acid (CB/
LA). Color codes followed the Pantone color code
adopting CYMK system referring to a Pantone
color bridge (Anonymous, 2005).
For previously known distribution, the databaseof Global Biodiversity Information Facility (GBIF,

http://www.gbif.org/occurrence, as of January 26,2019) was searched, and countries with occurrencesof the given species are shown with an asterisk (*).
Distributions known only from the literature areshown with double asterisks (**). Those with bothinformation are shown with triple asterisks (***).
All the specimens used in this study arehoused in the Department of Botany, National
Museum of Nature and Science (TNS), Tsukuba,
Japan.
DNA extraction and sequencing followed
Hosaka and Castellano (2008). Obtained isolates
will be deposited in NITE National Bioresource
Center (NBRC). The obtained DNA vouchers aredeposited in the Center for Molecular Biodiversity Research, National Museum of Nature andScience (Tsukuba, Ibaraki, Japan) and available
for molecular phylogenetic researches.
Descriptions
1.

Brunnipila clandestina (Bull.) Baral, Beih.

Yukito Tochihara et al.

48

Fig. 1. Brunnipila clandestina (TNS-F-81486). A. Fresh apothecia. B. Brunnipila clandestina with Capitotricha
rubi (arrow heads) on the same substrate (when fresh). C. Hairs with crystals. D. Vertical section of an apothecium showing the ectal and medullary excipulum. E. Ascus. F. Paraphysis. G. Ascospores. H. Ectal excipular
inner cells. Scales. A, B. 1 mm, C, E–H, 10 μm. D, 20 μm. C, E–H, mounted in CB/W; D, CB/LA.

Z. Mykol. 6: 49. 1985.

[Figs. 1, 2]

MycoBank. MB104069
Peziza clandestina Bull., Herb. Fr. Champ.,
Hist. Champ. Fr. (Paris) 1: 251. 1791.

Peziza clandestina var. clandestina Bull.,
Herb. Fr. Champ., Hist. Champ. Fr. (Paris) 1:
251. 1791.
Dasyscyphus clandestinus (Bull.) Fuckel,
Jb. nassau. Ver. Naturk. 23–24: 305. 1870
[1869–70].

Japanese Discomycetes (11)49

Fig. 2. Brunnipila clandestina (TNS-F-81486). A. Ascospores. B. Ectal excipular outer cells. C. Vertical section
through the margin, showing the hairs and ectal excipulum. D. Asci. E. Paraphyses. F. Hairs with and detached(left) crystals. G. Apothecium.

Lachnea clandestina (Bull.) P.Karst., Not.
Sällsk. Fauna et Fl. Fenn. Förh. 11: 250. 1870
[1871].
Erinella clandestina (Bull.) Quél., Enchir.
fung. (Paris): 302. 1886.
Lachnella clandestina (Bull.) W. Phillips,
Man. Brit. Discomyc. (London): 236. 1887.
Atractobolus clandestinus (Bull.) Kuntze,
Revis. gen. pl. (Leipzig) 3 (3): 445. 1898.
Peziza clandestina var. connivens Alb. &Schwein., Consp. fung. (Leipzig): 326. 1805.
Peziza clandestina var. patula Alb. & Schwein., Consp. fung. (Leipzig): 326. 1805.
Peziza clandestina var. discoidea Pers.,
Mycol. eur. (Erlanga) 1: 263. 1822.
Apothecia gregarious, superﬁcial, forming urce-

olate cup to almost ﬂat disc, 0.3–1 mm in diameter, stipitate, up to 1 mm high, dark brown(4635PC＝C13 M53 Y68 K40), externally covered with brown hairs mostly capitate. Disc concave, white to pale yellow (7499PC＝C1 M2
Y20 K0) when fresh and dry. Ectal excipulum
textura angularis to prismatica composed ofhyaline to pale brown (4655PC＝C6 M38 Y42
K18) cells; cells 3–17×3–12 μm, slightly thickwalled. Medullary excipulum textura intricata
of hyaline hyphae up to 2.5 μm wide. Hairs
straight, cylindrical with slightly rounded apices,
up to 120×3.5–5 μm, brown (471PC＝C5 M70
Y97 K20) with pale brown or hyaline apices,
totally granulate, thin-walled except for the apical cell slightly thick-walled, septate every
10–30 μm except for one or two apical cells with

Yukito Tochihara et al.

50

3–5 μm septation; apex crowned by crystal
masses easily detached in squash mounts. Asci
41–47×3.2–4.5 μm, 8-spored, cylindrical-clavate; apical pore blue in Melzerʼs reagent without
3% KOH pretreatment; croziers absent at thebasal septa; basal cells usually form more than
two asci. Ascospores 5.8–9×1.2–2 μm, ellipsoid
to fusiform, aseptate. Paraphyses straight, lanceolate, up to 5 μm wide, septate, exceeding theasci 20–30 μm.
Specimen examined. HOKKAIDO: TNS-F81486, Engaru-cho on dead twigs of Rubus
idaeus subsp. melanolasius. 2018-VII-14. coll. Y.
Tochihara.
Ecology. Saprophytic on dead twigs of Rubus
idaeus, sometimes on dead twigs or stems ofother plants.
Previously known distribution. Europe***,North America***, Kazakhstan*, Kyrgyzstan*,Tajikistan*, Morocco*, New Zealand*, Russia***(including Far East).
Notes. Baral and Krieglsteiner (1985) reported
B. clandestina coexisted with Capitotricha rubi(Bres.) Baral and Lachnum virgineum (Batsch) P.
Karst., on the same substrate (R. idaeus). We also
observed B. clandestina shared the twig of R.
idaeus with C. rubi in TNS-F-81486 (Fig. 1B).
Brunnipila clandestina, the type species ofBrunnipila Baral, is widely distributed corresponding to the distribution of its host plant
Rubus idaeus. In Japan, two occurrences of B.
clandestina (TNS-F-50486 and TNS-F-50462,
registered as ʻDasyscyphus clandestinusʼ) collected by Yoshio Otani, were found in GBIF.
However, as a result of our re-examination, TNSF-50486 and 50462 were found to be Trichopeziza sp. and Lachnum pseudocannabinum(Raitv.) Raitv. respectively. TNS-F-81486 istherefore the ﬁrst record of B. clandestina fromJapan.
2.

Brunnipila pseudocannabina (Raitv.) Tochihara, Sasagawa & Hosoya comb. nov.

[Figs. 3, 4]

MycoBank. MB829699

Dasyscyphus pseudocannabinus Raitv., Folia
Cryptog. Estonica 9: 6. 1977.
Lachnum pseudocannabinum (Raitv.) Raitv.,
Folia Cryptog. Estonica 17: 3. 1985.
Apothecia gregarious, superﬁcial, cup-shaped,
0.6–1.4 mm in diameter, stipitate, up to 1 mm
high, dark brown (Panton 4635PC＝C13 M53
Y68 K40), externally covered with capitate hairs.
Disc concave, white when fresh and dry. Ectal
excipulum textura prismatica to angularis, withcells 2–13×2–12 μm, hyaline, thick-walled up to
2 μm thick. Medullary excipulum textura intricata of hyaline hyphae up to 3 μm wide. Hairs
straight, cylindrical with slightly rounded apices,
up to 200×4–5 μm, brown (471PC＝C5 M70
Y97 K20) with pale brown or hyaline apices,
totally granulated, thin-walled, slightly thickwalled in one or two cells at the apices; apex
crowned by crystal masses easily detached insquash mounts. Asci 57–70×5.5–6 μm, 8-spored,
cylindrical-clavate; pore blue in Melzerʼs reagent
without 3% KOH pretreatment; croziers present
at the base. Ascospores 10–15×2–3 μm, slightly
long fusiform, sometimes irregularly curved,
aseptate or rarely one-septate, containing somesmall lipid bodies. Paraphyses straight, lanceolate with acute apices, up to 5 μm wide, septate,exceeding the asci for 25–30 μm.
Specimens examined. HOKKAIDO: TNS-F50462, Zenibako, Otaru-city on dead herb stems.
1967-VI-1. coll. Y. Otani, previously registeredas ʻDasyscyphus clandestinusʼ. TNS-F-81252,
Engaru-cho on dead stems of Filipendula camtschatica 2017-VII-12. coll. Y. Tochihara.
HONSHU: TNS-F-16690 (Culture NBRC
104374) and TNS-F-16691 (Culture NBRC
104382), Kosaka-machi, Akita Pref. on dead stemsof Fallopia, 2006-V-26. coll. R. Sasagawa. TNS-F17748 and 17751, Sugayu, Aomori-city, Aomori
Pref. 2005-VI-23. coll. T. Hosoya. TNS-F-18049,
Lake Towada, Towada-city, Aomori Pref. on deadherb stems. 2006-V-26. coll. T. Hosoya.
Ecology. Saprophytic on dead stems of tall
herbaceous plants.
Previously known distribution. KunashirJapanese Discomycetes (11)51

Fig. 3. Brunnipila pseudocannabina. A. Dried apothecia. B. Brunnipila pseudocannabina with Lachnum nudipes
dried (arrowhead) on the same substrate. C. Vertical section of an apothecium showing the ectal and medullary
excipulum. D. Ascus. E. Paraphysis. F. Hair with a crystal cap. G. Ectal excipular cells. H. Ascospore. A–F, H.
TNS-F-81252. G, TNS-F-16690. Scales. A, B, 1 mm. C, 20 μm. D–H, 10 μm. D–F, H, mounted in CB/W; C,
G, CB/LA.

Island (type locality) ***, Sakhalin**, Kamchatka *** (Raitviir, 1977; 1991).
Notes. TNS-F-16691 was examined in Hosoya etal. (2010) and determined as an unidentiﬁed lachnaceous species ʻLachnum sp. 4ʼ. Our re-examination revealed the six specimens of ʻLachnum sp. 4ʼ
were B. pseudocannabinum, in agreement with theoriginal description of Raitviir (1977).
Raitviir (1991) included L. pseudocannabinum

in section Brunnipila (Baral) Raitv., subgenus
Belonidium, genus Lachnum Retz. together with
L. cannabinum (＝currently B. cannabina), L.
clandestinum (＝currently B. clandestina) and L.
fuscidulum focusing on stipitate apothecia andbrown, hairs equipped with crystals. Since
molecular phylogenetic analysis (Hosoya et al.,2010) concluded that ʻLachnum sp. 4ʼ was positioned in the ʻBrunnipila cladeʼ, and we proposeYukito Tochihara et al.

52

Fig. 4. Brunnipila pseudocannabina (TNS-F-16624, 16771). A. Ascospores. B. Asci. C. Hairs with and detached(left) crystals. D. Vertical section through the margin, showing the hairs and ectal excipulum. E. Paraphyses. F.
Ectal excipular cells. G. Apothecium. Scales.

to transfer L. pseudocannabinum to the genus
Brunnipila. As we have not examined specimensof L. fuscidulum, we hesitate to propose a new
combination for L. fuscidulum.
Brunnipila pseudocannabina differs from B.
cannabina (Rehm) Raitv. & Järv, in having larger
asci ＞55 μm (Raitviir, 1991). Raitviir (1979,
1991) pointed out this fungus was endemic to the
Far East while B. cannabina was widely distributed throughout Russia and Europe. Brunnipila
pseudocannabina distribution probably includes
also the subarctic zone, from north of Central
Honshu to Kamchatka.

Brunnipila pseudocannabina usually shares
the same substrate with other lachnaceous fungi,
such as Lachnum nudipes (Fuckel) Nannf. (Fig.
3B), which is found on decaying stems of tallherbs. Species of Trichopezizella are similar to B.
pseudocannabina at ﬁrst glance, but differ inhaving sessile to substipitate apothecia andsmooth hairs lacking crystals.
For Japan, B. fuscescens (Pers.) Baral and B.
palearum (Desm.) Baral have been included in
the genus Brunnipila (Katsumoto, 2010). B.
pseudocannabina transfer is made in this study.

Japanese Discomycetes (11)53

Fig. 5. Incrucipulum capitatum (TNS-F-81420). A. Scattered apothecia on Quercus leaf. B. Dried apothecia. C.
Ascus. D. Paraphyses. E, F. Hairs. G. Ascospores. H. Ectal excipular cells. I. Ectal excipular cells with granulate surface. Scales. A, 1 cm. B, 1 mm. C–I, 10 μm. C–I, mounted in CB/W.

3.

Incrucipulum capitatum (Peck) Baral, Beih.
Z. Mykol. 6: 72. 1985.
[Figs. 5, 6]

MycoBank. MB104378
Peziza capitata Peck, Ann. Rep. N.Y. St.
Mus. nat. Hist. 30: 60. 1878. (1877).
Trichopeziza capitata (Peck) Sacc., Syll.
fung. (Abellini) 8: 417. 1889.
Dasyscyphus capitatus (Peck) Le Gal,
Revue Mycol., Paris 4: 29. 1939.
Lachnella capitata (Peck) Seaver, North

American Cup-fungi (Inoperculates) (New
York): 254. 1951.
Lachnum capitatum (Peck) Svrček, Česká
Mykol. 39 (4): 214. 1985.
Atractobolus scintillans (Massee) Kuntze,
Revis. gen. pl. (Leipzig) 3 (2): 446. 1898.
Dasyscyphus scintillans Massee [as ʻDasyscyphaʼ], Brit. Fung.-Fl. (London) 4: 328. 1895.
Apothecia scattered, superﬁcial, very minute, atﬁrst spherical and later urceolate, 0.1–0.2 mm indiameter, subsessile to sessile, up to 0.2 mm high,Yukito Tochihara et al.

54

Fig. 6. Incrucipulum capitatum (TNS-F-81420). A. Apothecium. B. Ascospores. C. Asci. D. Paraphyses. E.
Thick-walled hairs with crystals. F. Ectal exipular cells with granulate surface.

externally covered with short, white, stiff, capitate hairs. Disc concave, almost enclosed by anincurving margin when fresh and dry. Ectal
excipulum textura prismatica composed of
cubic cells like stone pavings with granulated
surface, 3–10×5–13 μm, hyaline, thick-walled
up to 2 μm wide; cells arranged in parallel rows.
Medullary excipulum textura intricata of hyaline hyphae up to 2 μm wide. Hairs straight,
cylindrical with rounded apices, 55–80×5.5–
7.5 μm, 2–3-septate, hyaline, totally granulate,thin to slightly thick-walled towards the base,very thick-walled up to 3 μm wide in one or twocells at the apices; apex crowned by crystal
masses easily detached in squash mounts. Asci

26.5–35×2–3.8 μm, 8-spored, cylindrical-clavate; pore blue in Melzerʼs reagent without 3%
KOH pretreatment; croziers absent at the basal
septa. Ascospores 4.5–10×1–1.2 μm, slightly
long fusiform, aseptate, containing some small
lipid bodies. Paraphyses straight, lanceolate, up
to 4 (–5) μm wide, aseptate, exceeding the asci
by 20 μm.
Specimen examined. HOKKAIDO: TNS-F81420, Jozankei, Sapporo-city on dead leaves ofQuercus crispula. 2018-VI-17. coll. T. Hosoya
and Y. Tochihara.
Ecology. Saprophytic on dead leaves of Quercus.
Previously known distribution. USA***,Japanese Discomycetes (11)Venezuela*, France*, Belgium*, Germany*,
Estonia***, UK***, Netherlands*, Lithuania*,
Armenia* (Dennis, 1949; Peck, 1878; Raitviir
1969; Seaver, 1951).
Notes. Incrucipulum capitatum is characterized by having lanceolate paraphyses, very-thickwalled and short hairs, and occurring on Quercus
leaves. Morphology and the host plant correspond descriptions by Peck (1878), Dennis(1949), Seaver (1951) and Raitviir (1969). Incrucipulum capitatum is newly documented from
Asia (including the Far East) in this study. In
Japan, Incrucipulum ciliare (Schrad.) Baral ispresent covering a wide area on Quercus leaves,
while I. capitatum is rare. Since the host Quercus
species are distributed in temperate zones, I. capitatum should be widely distributed in the temperate zones.
Acknowledgments
The authors are very thankful to an anonymous reviewer for valuable comments on the
manuscript.
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