Taxonomic study of the Japanese Dacrymycetes.

The class Dacrymycetes is a rather small group of brown-rot wood-decaying jelly fungi characterised by forked basidia and an orange to yellow gelatinous to cartilaginous fruit body. In Japan, dacrymycetous fungi had not been investigated for a long time, justifying a taxonomic re-examination. In the present study we attempted an investigation of the dacrymycetous fungal flora of Japan, and recognised 28 taxonomic entities, including five new taxa, i.e. Dacrymyces ancyleus, D. aureosporus, D. pinacearum, D. subarcticus and Dacryopinax sphenocarpa, and nine new records. Due to the present survey, the total number of dacrymycetous species recorded from Japan increased from 28 to 42. Of the newly described species, Dacrymyces ancyleus is characterised by recurved, cylindrical basidiocarps and hyphae with clamp connections. Dacrymyces aureosporus resembles D. chrysospermus, but differs in wall thickness of its marginal hyphae. Dacrymyces pinacearum and D. subarcticus represent new coelomycetous anamorphic species. Dacryopinax sphenocarpa has sharp, spathulate basidiocarps, and hyphae with clamp connections. Descriptions, illustrations and photographs of fruit bodies are presented with some taxonomic notes. Molecular phylogenetic analyses were conducted to verify the species identification, and the remaining problems in Dacrymycetes taxonomy are discussed based on these data.

INTRODUCTION

Dacrymycetes is a rather small class established by Doweld (2001) in the subphylum Agaricomycotina of Basidiomycota with one order, two families, nine genera and 101 species (Kirk et al. 2001, 2008, Hibbett et al. 2007). Dacrymycetes is characterised by forked (bifurcate) basidia, except in Dacrymyces unisporus. Their fruit bodies are gelatinous or cartilaginous, yellow to orange coloured, and vary in shape from thin membranous to pulvinate, spathulate and dendroid. In their hyphae, the dolipore-type septa are surrounded by parenthesomes without perforations (Wells 1994). All species belonging to this class are wood-decaying fungi that cause brown-rot (Oberwinkler 1993, Kirk et al. 2001). Recent molecular phylogenetic studies have shown that Dacrymycetes is a member of the Agaricomycotina clade with the Agaricomycetes and Tremellomycetes, and a sister group of Agaricomycetes (James et al. 2006, Hibbett 2006, Hibbett et al. 2007).

The only order in the class, Dacrymycetales, was established by Hennings (1898; as Dacryomycetinieae) composed of the single family Dacrymycetaceae, introduced by Schröter (1889; as Dacryomycetini) with several genera, until Jülich erected an additional family Cerinomycetaceae to accommodate the genus Cerinomyces, species of which produce resupinate basidiocarps (Jülich 1981, Kirk et al. 2001). In recent years, a taxonomic review of the genera in Dacrymycetaceae was conducted by McNabb (1964, 1965a, b, c, d, e, 1966, 1973). He re-examined the validity of described genera based on the type specimens and original descriptions, and finally recognised eight genera in the Dacrymycetaceae: Calocera, Cerinomyces, Dacrymyces, Dacryopinax, Ditiola, Femsjonia, Guepiniopsis and Heterotextus. He classified these genera mainly based on the external shapes of basidiocarps and the wall thickness of marginal hyphae in the sterile parts of basidiocarps (McNabb & Talbot 1973). The ninth genus, Dacryonaema, is monotypic and rarely reported (Nannfeldt 1947). The taxonomic history of dacrymycetalean genera has been reviewed by Oberwinkler (1993). Some studies discussing the phylogenetic relationship in this class have been published (Weiss & Oberwinkler 2001, Shirouzu et al. 2007).

The fungal floristic data of a certain region adds basic information on the classification and distribution of the species occurring in this region. Some mycologists reported the taxonomic study of dacrymycetous fungi distributed in a certain country, e.g. Reid (1974; UK) and Liu & Fan (1990; China). In Japan, 28 species representing seven genera have been recorded (Table 1), but systematic investigation of Japanese dacrymycetous fungi has not been conducted for 70 years since the works of Kobayasi (1939a, b). Although he described 14 new dacrymycetous species in Japan, several have remained doubtful (McNabb 1965a, b, e, 1973), as their holotypes have not been located. Given this situation, a re-examination of dacrymycetous fungi in Japan was urgently required.

Table 1

Species of Japanese Dacrymycetes.

Name	Literature	McNabb’s opinion3
Calocera alba Kobayasi	Kobayasi (1939b)	= C. cornea?
Calocera coralloides Kobayasi	Kobayasi (1939b)	= C. cornea?
Calocera cornea (Batsch) Fr.	Kobayasi (1939b)	distinct species
Calocera cornea forma glacilis Kobayasi	Kobayasi (1939b)	= C. cornea?
Calocera corniformis Kobayasi	Kobayasi (1939b)	= C. cornea?
Calocera furcata (Fr.) Fr.	Kobayasi (1939b); as Calocera flavida Lloyd	distinct species
Calocera viscosa (Pers.) Fr.	Kobayasi (1939b)	distinct species
Cerinomyces albosporus Boidin & Gilles	Maekawa (1986); as Cerinomyces aculeatus N. Maek.	–
Dacrymyces albidus Kobayasi	Kobayasi (1954)	distinct species?
Dacrymyces applanatus Kobayasi	Kobayasi (1939a)	distinct species?
Dacrymyces chrysocomus (Bull.) Tul.	Kobayasi (1939a)	distinct species
Dacrymyces chrysospermus Berk. & M.A. Curtis	Kobayasi (1939a); as Dacrymyces palmatus (Schwein.) Burt.,
	D. roseotinctus Lloyd, D. puniceus Kobayasi	distinct species
Dacrymyces kohyasanus Kobayasi	Kobayasi (1984)	–
Dacrymyces minor Peck	Kobayasi (1939a)	distinct species
Dacrymyces nikkomontanus Kobayasi	Kobayasi (1939a)	distinct species?
Dacrymyces pezizoides Kobayasi	Kobayasi (1939a)	= D. minutus?
Dacrymyces pulcher Kobayasi	Kobayasi (1939a)	distinct species?
Dacrymyces punctiformis Newhoff	Kobayasi (1939a)	distinct species
Dacrymyces san-augustinii Kobayasi	Kobayasi (1939a)	distinct species
Dacrymyces stillatus Nees	Kobayasi (1939a); as Dacrymyces deliquescens (Bull.) Duby	distinct species
Dacrymyces subalpinus Kobayasi	Kobayasi (1939a)	distinct species?
Dacrymyces tremellosus Kobayasi	Kobayasi (1939a)	= D. capitatus?
Dacryopinax imazekiana (Kobayasi) Lowy	Kobayasi (1939b); as Guepinia imazekiana Kobayasi	= D. dennisii?
Dacryopinax spathularia (Schwein.) G.W. Martin	Kobayasi (1939b); as Guepinia fissa Berk., G. spathularia (Schwein.) Fr.	distinct species
Femsjonia orientalis Kobayasi	Kobayasi (1939b)	= F. peziziformis?
Femsjonia peziziformis (Lèv.) P. Karst.	Kobayasi (1939b); as Femsjonia luteo-alba Fr.	distinct species
Guepiniopsis buccina (Pers.) L.L. Kenn.	Kobayasi (1939a); as Guepiniopsis merulinus (Pers.) Pat.	distinct species
Heterotextus alpinus (Tracy & Earle) Martin	Kobayasi (1939a); as Guepiniopsis alpinus (Tracy & Earle) Brasf.	distinct species

1 McNabb (1965a, b, e, 1973).

Bold: Species collected in this study.

In the present study we attempt to survey the dacrymycetous fungi of Japan. Fungal specimens collected from 2005 to 2008 were examined. Their descriptions and illustrations are presented with some taxonomic notes in this paper. Molecular phylogenetic analyses are also conducted using Japanese and foreign strains to verify species identification. Finally, the problems remaining in Dacrymycetes taxonomy are discussed based on these data.

MATERIALS AND METHODS

Fungal collections and morphological observations

To collect basidiocarps of dacrymycetous fungi, we selected approximately 50 sites in Japan with coniferous, deciduous broad-leaved and evergreen broad-leaved forests. The 23 major sites are shown in Fig. 1. These sites were sampled from 2005 to 2008. When sampling, basidiocarps were collected together with the substrata on which they occurred. The tree species or substrates were identified when possible. Collected samples were quickly air-dried, wrapped with paper and transported to the laboratory.

Fig. 1

Map of the Japanese Islands. Major sampling sites are shown as •.

In the laboratory, dried samples were soaked in distilled water to observe their morphological features. Basidiocarp shapes were observed with the naked eye and under a stereomicroscope. Basidiocarps were then sliced using a freezing microtome (RUB-2100, MC-802A; Yamato Kohki, Saitama, Japan) to a thickness of 10–40 μm to make preparations for microscopic examination. The microscopic features, such as basidia and basidiospores, were observed using a high-powered light microscope (150–1500×). Species identification was performed referring to the descriptions of Kobayasi (1939a), Olive (1944), McNabb (1964, 1965a, b, c, e, 1973), Oberwinkler & Tschen (1989) and Reid (1974). Identified specimens were deposited in the fungal herbarium of the National Museum of Nature and Science (TNS), Tsukuba, Ibaraki, Japan.

Fungal isolations

Pure cultures were established by means of single spore isolation. The established cultures were preserved in slants of 0.1 % cornmeal agar (0.2 % CMA; Nissui, Tokyo, Japan) + 1.25 % malt agar (2.5 % MA; Nissui, Tokyo, Japan) medium (0.2 % CMA 8.5 g, 2.5 % MA 22.5 g, yeast extract 1 g, distilled water 1 L), and deposited in the MAFF Genbank (National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan).

The colony growth rates and characteristics (texture, colour, etc.) were recorded based on 10 d old colonies grown in slants of 0.1 % CMA + 1.25 % MA medium at room temperature (c. 25 °C). Conidiogenesis was observed on colonies grown on 0.2 % CMA plates.

Molecular phylogenetic analyses

Fungal cultures for DNA sequencing were obtained from Japanese Dacrymycetes samples collected in this study, and from the Fungal Biodiversity Centre of the Centraalbureau voor Schimmelcultures (CBS) in Utrecht, The Netherlands. The list of samples used for phylogenetic analyses is shown in Table 2.

Table 2

Fungal samples used in molecular phylogenetic analysis.

Name	Sample no.	Specimen no.1	Culture no.2	GenBank accession no.3
Present study
Calocera cornea	HNo.267	TNS-F-15701	MAFF240116	AB299068
	HNo.358	TNS-F-15702	MAFF240117	AB299076
	HNo.376	TNS-F-15703	MAFF240118	AB299077
	HNo.452	TNS-F-21061	MAFF241186	AB472722†
	HNo.513	TNS-F-21065	MAFF241188	AB472725†
Calocera viscosa	HNo.175	TNS-F-15704	MAFF240119	AB299048
	HNo.466	TNS-F-15705	MAFF240120	AB299082
Cerinomyces albosporus	HNo.191	TNS-F-15706	MAFF240121	AB299050
Cerinomyces canadensis	HNo.199	TNS-F-21034	MAFF241162	AB472696
	HNo.208	TNS-F-21035	MAFF241163	AB472697
	HNo.216	TNS-F-21036	–	AB472698
	HNo.219	TNS-F-21037	MAFF241164	AB472699
Cerinomyces pallidus	HNo.505	TNS-F-21064	–	AB472724
Dacrymyces adpressus	HNo.355	TNS-F-21045	MAFF241172	AB472707
	HNo.554	TNS-F-21069	MAFF241191	AB472729
Dacrymyces ancyleus	HNo.382	TNS-F-21051	MAFF241177	AB472713
Dacrymyces aureosporus	HNo.215	TNS-F-15711	MAFF240126	AB299057
	HNo.354	TNS-F-21044	MAFF241171	AB472706
	HNo.385	TNS-F-21053	–	AB472715†
	HNo.486	TNS-F-15714	MAFF240129	AB299084
	HNo.665	TNS-F-21074	MAFF241195	AB472734†
Dacrymyces capitatus	HNo.182	TNS-F-15707	MAFF240122	AB299049
	HNo.212	TNS-F-15709	MAFF240124	AB299055
	HNo.471	TNS-F-21062	MAFF241187	AB472723†
	HNo.734	TNS-F-21077	MAFF241197	AB472737†
Dacrymyces capitatus (anam.)	HNo.181	TNS-F-21033	MAFF241161	AB472695
Dacrymyces chrysospermus	HNo.320	TNS-F-15712	MAFF240127	AB299073
	HNo.446	TNS-F-21060	MAFF241185	AB472721†
	HNo.468	TNS-F-15713	MAFF240128	AB299083
	HNo.620	TNS-F-21072	MAFF241194	AB472732†
Dacrymyces lacrymalis	HNo.209	TNS-F-15716	MAFF240131	AB299053
	HNo.235	TNS-F-15717	MAFF240132	AB299062
	HNo.243	TNS-F-21039	MAFF241166	AB472701
	HNo.261	TNS-F-15718	MAFF240133	AB299066
	HNo.271	TNS-F-21041	MAFF241168	AB472703
	HNo.277	TNS-F-21042	MAFF241169	AB472704
	HNo.279	TNS-F-21043	MAFF241170	AB472705
	HNo.281	TNS-F-15719	MAFF240134	AB299069
	HNo.563	TNS-F-21070	MAFF241192	AB472730
Dacrymyces lacrymalis (anam.)	HNo.250	TNS-F-21040	MAFF241167	AB472702
Dacrymyces microsporus	HNo.368	TNS-F-21049	MAFF241175	AB472711
	HNo.371	TNS-F-21050	MAFF241176	AB472712
	HNo.390	TNS-F-21054	MAFF241180	AB472716†
Dacrymyces minor	HNo.224	TNS-F-15720	MAFF240135	AB299059
	HNo.237	TNS-F-15721	MAFF240136	AB299063
Dacrymyces minutus	HNo.282	TNS-F-15722	MAFF240137	AB299070
	HNo.648	TNS-F-21073	–	AB472733†
Dacrymyces novae-zelandiae	HNo.225	TNS-F-21038	MAFF241165	AB472700
Dacrymyces pinacearum (anam.)	HNo.418	TNS-F-21056	MAFF241182	AB472718
Dacrymyces punctiformis	HNo.196	TNS-F-15723	MAFF240138	AB299052
	HNo.213	TNS-F-15724	MAFF240139	AB299056
	HNo.285	TNS-F-15725	MAFF240140	AB299071
Dacrymyces san-augustinii	HNo.441	TNS-F-15726	MAFF240141	AB299081
	HNo.666	TNS-F-21075	MAFF241196	AB472735†
Dacrymyces stillatus	HNo.233	TNS-F-15727	MAFF240142	AB299061
	HNo.256	TNS-F-15728	MAFF240144	AB299065
	HNo.383	TNS-F-21052	MAFF241178	AB472714
Dacrymyces stillatus (anam.)	HNo.252	TNS-F-15729	MAFF240143	AB299064
	HNo.411	TNS-F-21055	MAFF241181	AB472717
	HNo.421	TNS-F-21057	MAFF241183	AB472719
Dacrymyces subalpinus	HNo.228	TNS-F-15730	MAFF240145	AB299060
	HNo.570	TNS-F-21071	MAFF241193	AB472731†
Dacrymyces subarcticus (anam.)	HNo.544	TNS-F-21067	–	AB472727†
	HNo.722	TNS-F-21076	–	AB472736†
Dacrymyces unisporus	HNo.332	TNS-F-15731	MAFF240146	AB299074
Dacrymyces variisporus	HNo.263	TNS-F-15732	MAFF240147	AB299067
	HNo.300	TNS-F-15733	MAFF240148	AB299072
	HNo.352	TNS-F-15734	MAFF240149	AB299075
Dacryopinax spathularia	HNo.367	TNS-F-21048	MAFF241174	AB472710
	HNo.379	TNS-F-15735	MAFF240150	AB299078
	HNo.398	TNS-F-15736	MAFF240151	AB299079
Dacryopinax sphenocarpa	HNo.356	TNS-F-21046	MAFF241173	AB472708
	HNo.364	TNS-F-21047	–	AB472709
	HNo.430	TNS-F-21059	MAFF241184	AB472720
	HNo.534	TNS-F-21066	MAFF241189	AB472726†
	HNo.552	TNS-F-21068	MAFF241190	AB472728
Femsjonia peziziformis	HNo.439	TNS-F-15737	MAFF240152	AB299080
Guepiniopsis buccina	HNo.562	TNS-F-15738	MAFF240153	AB299085
Centraalbureau voor Schimmelcultures (CBS)
Dacrymycetes
Calocera cornea	CBS124.84	–	–	AB472738†
	CBS125.84	–	–	AB472739†
Calocera viscosa	CBS292.82	–	–	AB472740†
Dacrymyces capitatus	CBS293.82	–	–	AB472741†
Dacrymyces novae-zelandiae	CBS295.82	–	–	AB472742†
Dacrymyces stillatus	CBS296.82	–	–	AB472743†
Dacryopinax spathularia	CBS197.63	–	–	AB472744†
Guepiniopsis buccina	CBS297.82	–	–	AB472745†
DNA Data Bank of Japan (DDBJ)
Dacrymycetes
Calocera cornea	AFTOL-ID 438	–	–	AY701526
Calocera viscosa	AFTOL-ID 1679	–	–	DQ520102
Cerinomyces crustulinus	–	–	–	AY600248
Dacrymyces chrysospermus	–	–	–	AF287855
Dacrymyces stillatus	–	–	–	AF291309
Dacryomitra pussila	–	–	–	AJ406406
Dacryopinax spathularia	AFTOL-ID 454	–	–	AY701525
Dacryoscyphus chrysochilus	–	–	–	AY604567
Ditiola haasii	–	–	–	AF291314
Femsjonia peziziformis	–	–	–	AF291330
Guepiniopsis buccina	AFTOL-ID 888	–	–	AY745711
Agaricomycetes (out group)
Exidia uvapassa	AFTOL-ID 461	–	–	AY645056
Pseudohydnum gelatinosum	AFTOL-ID 1875	–	–	DQ520094

1 Herbarium of the National Museum of Nature and Science (TNS).

2 Culture collection of National Institute of Agrobiological Science (MAFF).

3 † Sequenced by Macrogen, Inc.

DNA was extracted from mycelia cultured on 2.5 % malt extract liquid medium following the modified CTAB method described by Matsuda & Hijii (1999). The 28S rDNA D1/D2 region was amplified with primers D1 (Peterson 2000) and NL4 (O’Donnell 1993). Polymerase chain reactions (PCR) were performed using a HotStarTaq Master Mix (Qiagen, Mississauga, Canada). Each PCR tube contained a 50 μL mixture (21 μL distilled water, 25 μL Master Mix, 3 μL template DNA, and 0.5 μL each primer; final 0.25 μM). Each DNA fragment was amplified using a PCR thermal cycler (Eppendorf Mastercycler Gradient; Eppendorf, Hamburg, Germany). The thermal cycling schedule was as follows: the first cycle consisted of 15 min at 94 °C, followed by 45 cycles of 30 s at 94 °C, 30 s at 58 °C for annealing, 1 min at 72 °C, and the final cycle of 10 min at 72 °C. The reaction mixture was then cooled at 4 °C for 5 min, and PCR products purified with a QiAquick PCR Purification Kit (Qiagen, Ontario, Canada).

Samples with a dagger (†) in Table 2 were sent to Macrogen, Inc. (Seoul, Korea) and sequenced. Sequencing reactions were performed in a MJ Research PTC-225 Eltier Thermal Cycler using an ABI PRISMR BigDyeTM Terminator Cycle Sequencing Kit with AmpliTaqR DNA polymerase (FS enzyme; PE Applied Biosystems, Foster City, CA, USA), following the protocols supplied by the manufacturer. Single-pass sequencing was performed on each template. The fluorescent-labelled fragments were purified from unincorporated terminators with an ethanol precipitation protocol. The samples were resuspended in distilled water and subjected to electrophoresis in an ABI 3730xl sequencer (PE Applied Biosystems).

Samples without a dagger (†) in Table 2 were sequenced in our laboratory. Sequencing reactions were performed using a BigDye Terminator Cycle Sequencing FS Ready Reaction Kit (PE Applied Biosystems) and an Eppendorf Mastercycler Gradient according to the manufacturer’s instructions. Sequencing reaction products were purified with a DyeEx Spin Kit (Qiagen, Mississauga, Canada), and directly sequenced using an ABI PRISM 377-18 DNA Sequencing System (PE Applied Biosystems). The sequences determined in this study were deposited in GenBank; their accession numbers are shown in Table 2. In addition to the sequences generated, 13 sequences accessed from GenBank belonging to Dacrymycetes and Agaricomycetes were included in the phylogenetic analysis (Table 2).

Preliminary multiple alignments of sequences were conducted using MAFFT v6 (Katoh et al. 2005, http://align.bmr.kyushu-u.ac.jp/mafft/software). Final alignments were manually adjusted. Alignment gaps were treated as missing data, and ambiguous positions were excluded from the analysis. Maximum-parsimony (MP) analyses were carried out using PAUP v4.0b10 (Swofford 2001). MP analyses with the heuristic search option using the tree-bisection-reconstruction (TBR) algorithm with 1 000 random sequence additions were performed to find the global optimum tree. All sites were treated as unordered and unweighted. To estimate clade support, the bootstrap (BS) procedure of Felsenstein (1985) was employed with 1 000 replicates in MP analyses and 100 replicates in the Maximum-likelihood (ML) analysis, with BS values higher than 50 % shown.

RESULTS

Taxonomy

As a result of the field work, about 600 samples were collected and 493 samples were identified to species level. There were 28 species representing 6 genera, of which 5 were new species, including 2 new anamorphic species, and 9 already-described species which were new records for Japan (Table 3). The descriptions, illustrations and photographs of these species are as follows.

Table 3

Species identified in this study.

Species	Literature	New record for Japan1
Calocera cornea (Batsch) Fr.	McNabb (1965a)	–
Calocera viscosa (Pers.) Fr.	McNabb (1965a)	–
Cerinomyces albosporus Boidin & Gilles	Boidin & Gilles (1986)	–
Cerinomyces canadensis (H.S. Jacks. & G.W. Martin) G.W. Martin	McNabb (1964)	°
Cerinomyces pallidus G.W. Martin	McNabb (1964)	°
Dacrymyces adpressus Grognot	McNabb (1973)	°
Dacrymyces ancyleus Shirouzu & Tokum.	This study	°
Dacrymyces aureosporus Shirouzu & Tokum.	This study	°
Dacrymyces capitatus Schwein.	McNabb (1973)	?
Dacrymyces chrysospermus Berk. & M.A. Curtis	McNabb (1973)	–
Dacrymyces dendrocalami Oberw.	Oberwinkler & Tschen (1989)	°
Dacrymyces lacrymalis (Pers.) Sommerf.	McNabb (1973)	°
Dacrymyces microsporus P. Karst.	McNabb (1973)	°
Dacrymyces minor Peck	McNabb (1973)	–
Dacrymyces minutus (L.S. Olive) McNabb	McNabb (1973)	?
Dacrymyces novae-zelandiae McNabb	McNabb (1973)	°
Dacrymyces pinacearum Shirouzu & Tokum.	This study	°
Dacrymyces punctiformis Neuhoff	Reid (1974)	–
Dacrymyces san-augustinii Kobayasi	Kobayasi (1939b)	–
Dacrymyces stillatus Nees	McNabb (1973)	–
Dacrymyces subalpinus Kobayasi	Kobayasi (1939b)	–
Dacrymyces subarcticus Shirouzu & Tokum.	This study	°
Dacrymyces unisporus (L.S. Olive) K. Wells	Olive (1944)	°
Dacrymyces variisporus McNabb	McNabb (1973)	°
Dacryopinax spathularia (Schwein.) G.W. Martin	McNabb (1965b)	–
Dacryopinax sphenocarpa Shirouzu & Tokum.	This study	°
Femsjonia peziziformis (Lèv) P. Karst.	McNabb (1965e)	–
Guepiniopsis buccina (Pers.) L.L. Kenn.	McNabb (1965c)	–

1 ° and – indicate new and existing Japanese records; ? unresolved.

Bold: newly described species in this study.

(Batsch) Fr., Stirp. Agric. Femis. 5: 67. 1827 — Fig. 2a–e, 14a–d

Fig. 2

a–e. Calocera cornea TNS-F-15701 (a, c–e; HNo.267), TNS-F-21061 (b, HNo.452). a. Simple cylindrical basidiocarps; b. palmate or dendroid basidiocarps; c. basidiospores; d. probasidia; e. developing basidium. — f–h. Calocera viscosa TNS-F-15705 (HNo.466). f. Basidiocarps; g. basidiospores; h. probasidium and developing basidium.

Fig. 14

Basidiocarps. a, b. Calocera cornea TNS-F-15701 (HNo.267); c, d. Calocera cornea TNS-F-21061 (HNo.452); e. Calocera viscosa TNS-F-15705 (HNo.466); f. Cerinomyces albosporus TNS-F-15706 (HNo.191); g. Cerinomyces canadensis TNS-F-21034 (HNo.199); h. Cerinomyces pallidus TNS-F-21064 (HNo.505); i. Dacrymyces adpressus TNS-F-21045 (HNo.355); j. Dacrymyces ancyleus TNS-F-21051 (HNo.382); k. Dacrymyces aureosporus TNS-F-15714 (HNo.486); l. Dacrymyces capitatus TNS-F-15708 (HNo.192); m. Dacrymyces chrysospermus TNS-F-15712 (HNo.320); n. Dacrymyces dendrocalami TNS-F-15715 (HNo.210); o. Dacrymyces lacrymalis TNS-F-15717 (HNo.235); p. Dacrymyces microsporus TNS-F-21050 (HNo.371). — Scale bars: a–d, f–p = 1 mm; e = 10 mm.

Basionym. Clavaria cornea Batsch, Elench. Fung. 1: 139. 1783.

For other synonyms see Reid (1974).

Basidiocarps scattered, cylindrical, subulate, simple, slightly branched, palmate or dendroid, white to yellow, soft-cartilaginous, 1–5 mm high, 1–2 mm diam. Structure showing in transverse section composed of a central core of compact parallel hyphae surrounded by a zone of loosely interwoven hyphae enclosed by the hymenium. Internal hyphae branched, thin- or thick-walled, septate, sub-hyaline, 2–4.5 μm diam, without clamp connections. Hymenium amphigenous. Probasidia cylindrical to clavate, pale yellow, 10–28 × 3.5–5.5 μm, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline, 7.5–12.5 × 3.5–6.5 μm (av. 10 × 4.5 μm; n = 20), 0–1-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 18 mm diam, velvety, pale orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5–6 × 3 μm.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of Quercus sp., 30 June 2006, T. Shirouzu, TNS-F-15702 (HNo.358); an unknown broad-leaved tree, 4 Oct. 2006, T. Shirouzu, TNS-F-21065 (HNo.513); Takaragaike, on dead branches of Quercus sp., 1 July 2006, T. Shirouzu, TNS-F-15703 (HNo.376); Nagano, Shioda, on dead branches of Pinus densiflora, 16 July 2006, T. Shirouzu, TNS-F-21061 (HNo.452); Wakayama, Mt Shirami, on dead branches of an unknown broad-leaved tree, 30 Apr. 2006, T. Shirouzu, TNS-F-15701 (HNo.267), culture MAFF240116.

Notes — This species is characterised by hyphae without clamp connections, comparatively small, simple basidiocarps and 1-septate basidiospores. We found two types of basidiocarps for this species: simple or slightly branched basidiocarps and palmate or dendroid basidiocarps. As in earlier studies (McNabb 1965a, Reid 1974, Tubaki & Hosoya 1987), this species was frequently found on woody materials of broad-leaved trees. This was previously reported in Japan (Kobayasi 1939b, Tubaki & Hosoya 1987).

(Pers.) Fr., Stirp. Agric. Femis. 5: 67. 1827 — Fig. 2f–h, 14e

Basionym. Clavaria viscosa Pers., Neues Mag. Bot. 1: 117. 1794.

For other synonyms see McNabb (1965a).

Basidiocarps scattered, dendroid composed of cylindrical dichotomous branches, yellow to orange, white near the base, soft-cartilaginous, 10–30 mm high, 1–2 mm diam. Structure showing in transverse section composed of a central core of compact parallel hyphae surrounded by a zone of loosely interwoven hyphae enclosed by the hymenium. Internal hyphae branched, thin-walled, septate, pale yellow, 2–5 μm diam, without clamp connections. Hymenium amphigenous. Probasidia cylindrical, yellow, 25–37 × 3–5 μm, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, pale yellow, 7.5–15 × 3.5–6.5 μm (av. 10 × 4.5 μm; n = 20), 0–1-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 5 mm diam, velvety to lanose, white-yellow. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Nagano, Sugadairakougen, on litter of Abies veitchii Lindl., 10 Aug. 2005, T. Shirouzu, TNS-F-15704 (HNo.175), culture MAFF240119; Saitama, Chichibu, on dead branches of an unknown conifer, 20 July 2006, Y. Takahashi, TNS-F-15705 (HNo.466).

Notes — This species is characterised by hyphae without clamp connections and large, dichotomously branched orange-coloured basidiocarps. In contrast to C. cornea, C. viscosa has been frequently found on woody materials of coniferous trees (McNabb 1965a, Reid 1974). In Japan, this has been also reported from coniferous wood (Kobayasi 1939b).

Boidin & Gilles, Bull. Trimestriel Soc. Mycol. France 102: 318. 1986 — Fig. 3a–e, 14f

Fig. 3

a–e. Cerinomyces albosporus TNS-F-15706 (HNo.191). a. Basidiocarp; b. basidiospores; c. probasidia; d. developing basidia; e. hyphal peg. — f–i. Cerinomyces canadensis TNS-F-21034 (HNo.199). f. Basidiocarp; g. basidiospores; h. probasidia; i. developing basidium. — j–m. Cerinomyces pallidus TNS-F-21064 (HNo.505). j. Basidiocarps; k. basidiospores; l. probasidia; m. developing basidium.

= Cerinomyces aculeatus N. Maek., Canad. J. Bot. 65: 583. 1987.

Basidiocarps resupinate, grey-orange, velvety, 3–10 mm long, 2–10 mm wide, 150–350 μm thick, covered with hymenium. Internal hyphae branched, thin-walled, septate, sub-hyaline, 2–3 μm diam, with clamp connections. Hyphal pegs acute to cylindrical, composed of septate, thin-walled hyphae with clamp connections, up to 180 μm above the hymenium, 30–80 μm diam. Probasidia cylindrical to clavate, pale yellow, 17.5–23.5 × 3–5 μm, with a basal clamp connection, becoming bifurcate. Basidiospores cylindrical to reniform, with an apiculum at the base, thin-walled, hyaline, 15–21 × 5–8.5 μm (av. 15.5 × 6.5 μm; n = 10), 0-septate, germination via germ tubes.

Cultural characteristics — Colonies attaining about 4 mm diam, wet, white. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6 × 3 μm.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of Pinus densiflora, 20 Apr. 2006, T. Shirouzu, TNS-F-15706 (HNo.191), culture MAFF240121; Nagano, Sugadairakougen, on dead branches of P. densiflora, 18 Aug. 2006, T. Shirouzu, TNS-F-21063 (HNo.478).

Notes — Cerinomyces albosporus is characterised by hyphae with clamp connections, presence of hyphal pegs and cylindrical to reniform basidiospores. This species has been reported in Japan by Maekawa (1987) as C. aculeatus. All the Japanese materials have been found on wood of Pinus densiflora.

(H.S. Jacks. & G.W. Martin) G.W. Martin, Mycologia 41: 85. 1949 — Fig. 3f–i, 14g

Basionym. Ceracea canadensis H.S. Jacks. & G.W. Martin, Mycologia 32: 693. 1940.

Basidiocarps resupinate, yellow, soft-waxy, 2–5 mm long, 2–3 mm wide, 100–380 μm thick, covered with hymenium. Internal hyphae branched, thin- or thick-walled, septate, sub-hyaline, 2–5 μm diam, with clamp connections. Probasidia cylindrical to clavate, pale yellow, 28.5–32.5 × 4–5 μm, with a basal clamp connection, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline, 8–13 × 3.5–5.5 μm (av. 10 × 4.5 μm; n = 15), 0–1-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 6 mm diam, velvety, white. Conidiogenesis not observed.

Specimens examined. Japan, Kyoto, Midorogaike, on dead branches of Rhododendron macrosepalum, 21 Apr. 2006, T. Shirouzu, TNS-F-21036 (HNo.216); Mt Daimonji, on dead branches of Pinus densiflora, 20 Apr. 2006, T. Shirouzu, TNS-F-21034 (HNo.199), culture MAFF241162; Mt Kiyomizu, on dead branches of Castanopsis cuspidata, 22 Aug. 2006, T. Shirouzu, TNS-F-21037 (HNo.219); Takaragaike, on dead branches of Clethra barbinervis, 21 Apr. 2006, T. Shirouzu, TNS-F-21035 (HNo.208).

Notes — Cerinomyces canadensis is characterised by yellow-coloured basidiocarps, absence of dikaryophyses and hyphal pegs, and relatively large, 0–1-septate basidiospores. This is the first report from Japan.

G.W. Martin, Mycologia 41: 83. 1949 — Fig. 3j–m, 14h

Basidiocarps resupinate, grey-olive to amber, soft-waxy, 5–20 mm long, 2–5 mm wide, 120–160 μm thick, covered with hymenium. Internal hyphae branched, thin-walled, septate, sub-hyaline, 2–5 μm diam, with clamp connections. Probasidia cylindrical to clavate, pale yellow, 22–29 × 4–6 μm, with a basal clamp connection, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, pale yellow, 7–12 × 3.5–5.5 μm (av. 9 × 4.5 μm; n = 15), 0-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 8 mm diam, velvety, white. Conidiogenesis not observed.

Specimen examined. Japan, Nagano, Shioda, on dead branches of Pinus densiflora, 27 Sept. 2006, T. Shirouzu, TNS-F-21064 (HNo.505).

Notes — Cerinomyces pallidus is characterised by olive to subdued-yellow basidiocarps, absence of dikaryophyses and hyphal pegs, and relatively small, 0-septate basidiospores. Compared with the description by McNabb (1964), the size of basidiospores was slightly larger in our specimens. This is the first report from Japan.

Grognot, Pl. Crypt. Sâone-et-Loire: 200. 1863 — Fig. 4a–e, 14i

Fig. 4

a–e. Dacrymyces adpressus TNS-F-21045 (HNo.355). a. Basidiocarps; b. basidiospores; c. probasidium; d. developing basidium; e. dikaryophyses. — f–i. Dacrymyces ancyleus TNS-F-21051 (HNo.382). f. Basidiocarps; g. basidiospores; h. probasidium; i. developing basidium.

For synonyms see McNabb (1973).

Basidiocarps scattered, pulvinate to cerebriform, sometimes applanate, sessile, orange to amber, soft-cartilaginous to firm-gelatinous, 1–3 mm high, 1–5 mm diam. Sterile parts of basidiocarps covered with simple or branched cylindrical to clavate, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, sub-hyaline, 2–6 μm diam, with clamp connections. Hymenium limited to the superior surface of the basidiocarp. Probasidia cylindrical to clavate, thick-walled, pale yellow, 45–73.5 × 5–8 μm, with a basal clamp connection, becoming bifurcate. Dikaryophyses simple or branched, septate, thin-walled, pale yellow, 40–90 × 3–4.5 μm. Basidiospores cylindrical to reniform, with an apiculum at the base, thin- or thick-walled, pale yellow, 15–24 × 6–10 μm (av. 19 × 8 μm; n = 20), 3-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 10 mm diam, velvety, white-yellow. Conidiogenesis not observed.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of an unknown broad-leaved tree, 30 June 2006, T. Shirouzu, TNS-F-21045 (HNo.355), culture MAFF241172; Wakayama, Mt Shirami, on dead branches of an unknown woody plant, 12 Oct. 2006, T. Shirouzu, TNS-F-21069 (HNo.554).

Notes — Dacrymyces adpressus is characterised by pulvinate, brown to amber-coloured basidiocarps, hyphae with clamp connections, simple dikaryophyses and relatively large, thin- or thick-walled, 3-septate basidiospores. This species sometimes produces applanate basidiocarps and it may be confused with Cerinomyces species. Dacrymyces adpressus Kobayasi was described in Japan is a later homonym, and a different species from D. adpressus Grognot (McNabb 1973), so this is the first record from Japan.

Shirouzu & Tokum., sp. nov. — MycoBank MB514036; Fig. 4f–i, 14j

Basidiocarpia sparsa, cylindrica, stipitata cum pileo subgloboso vel cylindrico ancyleo, flavida, cartilaginea vel gelatinosa, 1–2 mm alta, 0.5 mm lata. Hyphae interaneae ramosae, tenuitunicatae vel crassitunicatae, gelatinosae, hyalinae, 2–6 μm latae, cum colligationibus unciformibus. Probasidia cylindrica vel clavata, flavida, 30–45 × 4.5–10 μm, bifurcatascentia. Basidiosporae subglobosae vel reniformae, tenuitunicatae, sub-hyalinae, 10.5–19.5 × 4–9 μm, 0–1-septatae.

Etymology. Named after its recurved basidiocarps.

Basidiocarps scattered, cylindrical, stipitate, bearing a subglobose to cylindrical, recurved pileus, yellow, soft-cartilaginous to firm-gelatinous, 1–2 mm high, 0.5 mm diam. Internal hyphae branched, thin- or thick-walled, gelatinous, septate, hyaline, 2–6 μm diam, with clamp connections. Hymenium limited to surface of the pileus. Probasidia cylindrical to clavate, pale yellow, 30–45 × 4.5–10 μm, with a basal clamp connection, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline, 10.5–19.5 × 4–9 μm (av. 15.5 × 7.5 μm; n = 20), 0–1-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 4 mm diam, velvety, white. Conidiogenesis not observed.

Specimen examined. Japan, Kyoto, Takaragaike, on dead branches of Rhododendron sp., 1 July 2006, T. Shirouzu, holotype TNS-F-21051 (HNo.382), culture ex-type MAFF241177.

Notes — Dacrymyces ancyleus is characterised by basidiocarps that are recurved, cylindrical or stipitate with a subglobose pileus, hyphae with clamp connections and 0–1-septate basidiospores. This fungus is likely to be included in the genus Calocera based on the cylindrical basidiocarps; however, the internal hyphal structure of the basidiocarp could not be divided into three zones, differing from Calocera species. Furthermore, this fungus is not a member of Dacryopinax since the hymenium is not amphigenous and cortical hairs at the sterile surface could not be recognised; therefore, we regard this dacrymycetous fungus to belong to the genus Dacrymyces.

Dacrymyces microsporus is similar in basidiocarp and basidiospore shape to D. ancyleus, but the hyphae composing basidiocarps of D. microsporus do not have clamp connections. Dacrymyces flabelliformis is also similar to D. ancyleus in basidiocarp shape and by having clamp connections on hyphae (Burdsall & Laursen 2004), but D. flabelliformis is different from this fungus in basidiospore size (12.5–14 × 5–6 μm) and the septal number (3-septate); therefore, we consider this fungus to be a new species of Dacrymyces. In molecular phylogenetic analyses (Fig. 16), this species represented a monophyletic group with Dacryopinax sphenocarpa, but the phylogenetic position in the Dacrymycetes lineage was uncertain.

Fig. 16

Most parsimonious tree of Dacrymycetes using 28S rDNA D1/D2 region sequences. One of the 24 most parsimonious trees. Length = 901, CI = 0.3563, RI = 0.8137. Branch with asterisk (*) collapse in the strict consensus tree of all most parsimonious trees. Numbers above the node or to the left of slashes (/) indicate support above 50 % in 1 000 bootstrap replicates with parsimony analyses. Numbers above the node or to the right of slashes (/) indicate support above 50 % in 100 bootstrap replicates with likelihood analyses. Bold nodes are supported more than 80 % MP and 80 % ML bootstrap values. Newly described species in this study are shown in bold. TreeBASE SN4250.

Shirouzu & Tokum., sp. nov. — MycoBank MB514037; Fig. 5a–f, 14k

Fig. 5

a–f. Dacrymyces aureosporus TNS-F-15714 (HNo.486). a. Basidiocarps; b. basidiospores; c. probasidium; d. basidium and probasidium; e. dikaryophysis; f. marginal hyphae. — g–j. Dacrymyces capitatus TNS-F-15708 (HNo.192). g. Basidiocarps; h. basidiospores; i. probasidia and basidia; j. marginal hyphae.

Basidiocarpia sparsa vel gregaria, turbinata vel cerebriformia, sessilia vel stipitata cum pileo semigloboso rugoso vel convoluto, aurantiaca, gelatinosa, 1–3 mm alta, 1–6 mm lata. Pili corticales steriles cylindrici vel clavati, tenuitunicati. Hyphae interaneae ramosae, tenuitunicatae, gelatinosae, hyalinae, 2–3 μm latae. Probasidia cylindrica, sub-hyalina, 46.5–80 × 5.5–8 μm, bifurcatascentia. Basidiosporae cylindricae vel allantoideae, tenuitunicatae, flavidae, 17–26 × 6.5–11 μm, 7-septatae.

Etymology. Named after its yellow basidiospores.

Basidiocarps scattered or gregarious, sometimes coalesced, turbinate to cerebriform, sessile or stipitate bearing a rugose to convolute semiglobose pileus, pale orange, firm-gelatinous, 1–3 mm high, 1–6 mm diam. Sterile parts of basidiocarps covered with simple cylindrical to clavate, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–3 μm diam, without clamp connections. Hymenium limited to the upper surface of the pileus. Probasidia cylindrical, sub-hyaline, 46.5–80 × 5.5–8 μm, becoming bifurcate. Basidiospores cylindrical to allantoid, with an apiculum at the base, thin-walled, yellow, 17–26 × 6.5–11 μm (av. 21.5 × 8.5 μm; n = 20), 7-septate, germination via the production of conidia and germ tubes.

Culture characteristics — Colonies attaining about 6 mm diam, velvety, pale orange to orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 4 μm.

Specimens examined. Japan, Kyoto, Midorogaike, on dead branches of Quercus serrata, 21 Apr. 2006, T. Shirouzu, TNS-F-15711 (HNo.215); on dead branches of Cryptomeria japonica, 1 July 2006, T. Shirouzu, TNS-F-21053 (HNo.385); Mt Daimonji, on dead branches of an unknown broad-leaved tree, 30 June 2006, T. Shirouzu, TNS-F-21044 (HNo.354); Nagano, Kijimadaira, on a fallen tree of Fagus crenata, 2 Sept. 2006, T. Shirouzu, holotype TNS-F-15714 (HNo.486), culture ex-type MAFF240129; Okinawa, Iriomote Island, on a fallen tree of an unknown broad-leaved tree, 9 June 2007, T. Shirouzu, TNS-F-21074 (HNo.665).

Notes — Dacrymyces aureosporus is characterised by turbinate to cerebriform basidiocarps, hyphae without clamp connections, thin-walled, marginal hypha and multi-septate basidiospores. This fungus resembles D. chrysospermus in morphological features, except for the absence of thick-walled terminal cells on the sterile surface of the basidiocarps. Dacrymyces aureosporus also produces terminal cells but its walls do not become thick. Kobayasi (1939a) reported a similar dacrymycetous fungus to D. palmatus, which is a synonym of D. chrysospermus, and the terminal cells of the fungus are also thin-walled. In molecular phylogenetic analyses, D. chrysospermus and D. aureosporus were clearly separated (Fig. 16). We considered this to be a new Dacrymyces species closely related to D. chrysospermus.

Schwein., Trans. Amer. Philos. Soc., Ser. II, 4: 186. 1832 — Fig. 5g–j, 14l

For synonyms see McNabb (1973) and Reid (1974).

Basidiocarps scattered, turbinate to stoutly cylindrical, stipitate bearing a concave or rugose semiglobose pileus, yellow to orange, firm-gelatinous, sometimes white tomentose at the base, 1.5–2 mm high, 2–4 mm diam. Sterile parts of basidiocarps densely covered with simple or branched, cylindrical to clavate, septate, hyaline, thin- or thick-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–3.5 μm diam, without clamp connections. Hymenium limited to the upper surface of the pileus. Probasidia cylindrical to clavate, pale yellow, 25–42.5 × 3–6 μm, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline to pale yellow, 10–15 × 5–7.5 μm (av. 13 × 6.5 μm; n = 20), 0–3-septate, germination via conidia and germ tubes. Anamorphic fruit bodies sometimes present with basidiocarps, pulvinate, sessile, orange, gelatinous, 1 mm high, 1–3 mm diam. Conidia holoblastic, cylindrical to subglobose, with a separation scar at the base, thin- or thick-walled, 20–30 × 7.5–10 μm, 0–1-septate.

Culture characteristics — Colonies attaining about 4 mm diam, velvety, yellow to orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of Pinus densiflora, 29 Mar. 2006, T. Shirouzu, TNS-F-21033 (HNo.181; with anamorph stage); 20 Apr. 2006, T. Shirouzu, TNS-F-15708 (HNo.192), culture MAFF240123; TNS-F-15707 (HNo.182); Takaragaike, on dead branches of Clethra barbinervi, 21 Apr. 2006, T. Shirouzu, TNS-F-15709 (HNo.212); Nagano, Sugadairakougen, on dead branches of Picea sp., 19 July 2006, T. Shirouzu, TNS-F-21062 (HNo.471); on dead branches of Pinus densiflora, 7 Aug. 2007, T. Shirouzu, TNS-F-21077 (HNo.734).

Notes — This species is characterised by turbinate or sub-stipitate basidiocarps, hyphae without clamp connections, densely arranged thin- or thick-walled marginal hyphae, and thin-walled, 3-septate basidiospores. In this study, we collected material with anamorphic fruit bodies. Oberwinkler (1993) also reported that the species produced conidium-like diaspores by hyphal phragmentation. According to the taxonomic opinion of McNabb (1973), Dacrymyces capitatus might be reported in Japan as D. tremellosus (Kobayasi 1939a).

Berk. & M.A. Curtis, Grevillea 2: 20. 1873 — Fig. 6a–e, 14m

Fig. 6

a–e. Dacrymyces chrysospermus TNS-F-15712 (HNo.320). a. Basi-diocarps; b. basidiospores; c. probasidium; d. developing basidium; e. marginal hyphae. — f–j. Dacrymyces dendrocalami TNS-F-15715 (HNo.210). f. Basidiocarps; g. basidiospores; h. probasidium and dikaryophysis; i. basidium; j. dikaryophysis. — k–n. Dacrymyces lacrymalis TNS-F-15717 (HNo.235). k. Basidiocarps; l. basidiospores; m. developing basidium; n. pro-basidium and basidium.

For synonyms see McNabb (1973) and Reid (1974).

Basidiocarps scattered or gregarious, sometimes coalesced, turbinate to cerebriform, sessile or stipitate bearing a rugose to convolute semiglobose pileus, orange, firm-gelatinous, 2–5 mm high, 6–13 mm diam. Sterile parts of basidiocarps covered with simple cylindrical to clavate, septate, 2–3-celled, hyaline, conspicuously thick-walled terminal cells, 25–50 × 7.5–16 μm. Internal hyphae branched, thin-walled, gelatinous, septate, pale yellow, 2–3 μm diam, without clamp connections. Hymenium limited to upper surface of the pileus. Probasidia cylindrical to clavate, yellow to orange, 30–48 × 5–8 μm, becoming bifurcate. Basidiospores cylindrical to curved-cylindrical, with an apiculum at the base, thin-walled, yellow, 12–23 × 5–10 μm (av. 18 × 7 μm; n = 20), 3–7-septate, germinated by production of conidia and germ tubes.

Culture characteristics — Colonies of the primary mycelium attaining about 10 mm diam, velvety, orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6 × 2 μm.

Specimens examined. Japan, Kagoshima, Amami Island, on dead branches of Pinus luchuensis, 18 Jan. 2007, T. Shirouzu, TNS-F-21072 (HNo.620); Nagano, Sugadairakougen, on dead branches of Abies sp., 22 June 2006, T. Shirouzu, TNS-F-15712 (HNo.320), culture MAFF240127; a log of Larix kaempferi, 16 July 2006, T. Shirouzu, TNS-F-21060 (HNo.446); a standing log of L. kaempferi, 19 July 2006, T. Shirouzu, TNS-F-15713 (HNo.468).

Notes — This species is characterised by turbinate to cerebriform basidiocarps, hyphae without clamp connections, thick-walled terminal cells and multi-septate, cylindrical to curved, thin-walled basidiospores. It has previously been recorded in Japan as Dacrymyces palmatus, D. roseotinctus and D. puniceus (Kobayasi 1939a).

Oberw., Trans. Mycol. Soc. Japan 30: 350. 1989 — Fig. 6f–j, 14n

Basidiocarps scattered, pustulate to pulvinate, sessile, pale white to pale orange, firm-gelatinous, 0.2–0.5 mm high, 1–3 mm diam. Sterile parts of basidiocarps covered with simple or branched, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, 2–3 μm diam, without clamp connections. Hymenium limited to the superior surface of the basidiocarp. Probasidia cylindrical to clavate, thin- or thick-walled, hyaline to pale yellow, 24–37.5 × 5–7.5 μm, becoming bifurcate. Dikaryophyses simple or branched, septate, thin-walled, sub-hyaline, 2–3 μm diam. Basidiospores naviculate, straight or curved, with an apiculum at the base, thick-walled, sub-hyaline, 16–22.5 × 6–8 μm (av. 20 × 7 μm; n = 20), 0–3-septate, germination via germ tubes.

Culture characteristics — Culture not obtained.

Specimens examined. Japan, Kyoto, Takaragaike, on dead branches of an unknown broad-leaved tree, 21 Apr. 2006, T. Shirouzu, TNS-F-15715 (HNo.210); Nara, Mt Tamaki, on dead branches of Parabenzoin trilobum, 7 July 2006, T. Shirouzu, TNS-F-21058 (HNo.426).

Notes — Dacrymyces dendrocalami is characterised by pustulate to pulvinate, sessile basidiocarps, hyphae without clamp connections, and straight- or curved-naviculate, thick-walled basidiospores. This species has been reported only once from Taiwan (Oberwinkler & Tschen 1989). This is the second record for this species, and the first from Japan.

(Pers.) Sommerf., Suppl. Fl. Lapp. (Oslo) no. 1753: 308. 1826 — Fig. 6k–n, 14o

Basionym. Tremella lacrymalis Pers., Syn. Meth. Fung. 2: 628. 1801.

For other synonyms see McNabb (1973).

Basidiocarps scattered or gregarious, sometimes coalesced, pulvinate or irregularly discoid, applanate, gyrose or centrally depressed, sessile or sub-stipitate, yellow, firm-gelatinous, 0.5–2 mm high, 1–4 mm diam. Sterile parts of basidiocarps covered with simple or branched, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–3 μm diam, without clamp connections. Hymenium limited to the upper surface of the basidiocarp. Probasidia cylindrical to clavate, sub-hyaline to pale yellow, 20.5–30 × 3.5–5 μm, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline to pale yellow, 9.5–15 × 3.5–6 μm (av. 11.5 × 5 μm; n = 20), 0–3-septate, germinated by production of conidia and germ tubes. Anamorphic fruit bodies sometimes present with basidiocarps, pulvinate, sessile, yellow, gelatinous, 1 mm high, 1–3 mm diam. Conidia holoblastic, cylindrical to subglobose, with a separation scar at the base, thin-walled, 5–10 × 3–5 μm, 0–1-septate.

Culture characteristics — Colonies attaining about 8 mm diam, velvety, yellow. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Kyoto, Takaragaike, on dead branches of an unknown broad-leaved tree, 21 Apr. 2006, T. Shirouzu, TNS-F-15716 (HNo.209); Nagano, Sugadairakougen, on dead branches of Abies sp., 8 May 2006, T. Shirouzu, TNS-F-21041 (HNo.271); on dead branches of Alnus japonica, 14 May 2006, D. Hirose, TNS-F-15719 (HNo.281); on dead branches of Larix sp., 8 May 2006, T. Shirouzu, TNS-F-21042 (HNo.277); L. kaempferi, 28 May 2006, T. Shirouzu, TNS-F-21043 (HNo.279); Nara, Mt Tamaki, on dead branches of Fagus crenata, Nara, 27 Apr. 2006, T. Shirouzu, TNS-F-15718 (HNo.261); on dead branches of an unknown broad-leaved tree, 13 Oct. 2006, T. Shirouzu, TNS-F-21070 (HNo.563); Oodaigahara, on dead branches of F. crenata, 24 Apr. 2006, T. Shirouzu, TNS-F-15717 (HNo.235), culture MAFF240132; on dead branches of an unknown conifer, 24 Apr. 2006, T. Shirouzu, TNS-F-21040 (HNo. 250; with anamorph stage); on dead branches of F. crenata, 24 Apr. 2006, T. Shirouzu, TNS-F-21039 (HNo.243).

Notes — Dacrymyces lacrymalis is characterised by irregularly discoid, gyrose, sessile basidiocarps, hyphae without clamp connections and thin-walled, 3-septate basidiospores. Although McNabb (1973) did not report the anamorphic stage of this species, anamorphic fruit bodies bearing holoblastic conidia were observed together with basidiocarps in this study. According to the result of this study and McNabb (1973), D. lacrymalis has been frequently found on woody substrata of broad-leaved trees. This record is the first for Japan.

P. Karst., Bidrag Kannedom Finlands Natur Folk 48: 459. 1889 — Fig. 7a–c, 14p

Fig. 7

a–c. Dacrymyces microsporus TNS-F-21050 (HNo.371). a. Basidiocarps; b. basidiospores; c. probasidia. — d–g. Dacrymyces minor TNS-F-15720 (HNo.224). d. Basidiocarps; e. basidiospores; f. probasidium; g. developing basidium. — h–m. Dacrymyces minutus TNS-F-15722 (HNo.282). h. Basidiocarps; i. basidiospores; j. probasidium; k. developing basidium; l. dikaryophysis; m. marginal hyphae.

Basidiocarps scattered or gregarious, sometimes coalesced, turbinate to stoutly cylindrical, stipitate bearing a concave or rugose semiglobose pileus, pale yellow, soft-cartilaginous to firm-gelatinous, 1–1.5 mm high, 1–2 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled, marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–4 μm diam, without clamp connections. Hymenium limited to upper surface of the pileus. Probasidia cylindrical to clavate, sub-hyaline, 27.5–42.5 × 3.5–4.5 μm, becoming bifurcate. Basidiospores reniform, with an apiculum at the base, thin-walled, sub-hyaline, 10–14 × 3.5–6.5 μm (av. 11.5 × 5 μm; n = 20), 0–1-septate, germination by means of conidial production and germ tubes.

Culture characteristics — Colonies of the primary mycelium attaining about 7 mm diam, wet, yellow. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6 × 3 μm.

Specimens examined. Japan, Kyoto, Mt Kiyomizu, on dead branches of an unknown broad-leaved tree, 2 July 2007, T. Shirouzu, TNS-F-21054 (HNo.390); Takaragaike, on a fallen tree of Quercus sp., 1 July 2006, T. Shirouzu, TNS-F-21050 (HNo.371), culture MAFF241176; Wakayama, Mt Nachi, on dead branches of an unknown woody plant, 6 July 2006, T. Shirouzu, TNS-F-21049 (HNo.368).

Notes — Dacrymyces microspores is characterised by basiodiocarps that are turbinate to stipitate, bearing a concave or rugose, semiglobose pileus, hyphae without clamp connections and thin-walled, 0–1-septate basidiospores. This is the first record from Japan.

Peck, Ann. Rep. N. Y. State Mus. 30: 49. 1878 — Fig. 7d–g, 15a

For synonyms see McNabb (1973).

Basidiocarps scattered, pustulate to pulvinate, sessile or sub-stipitate, pale orange, firm-gelatinous, 0.5 mm high, 1–2 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled, marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–3 μm diam, without clamp connections. Hymenium limited to superior surface of the basidiocarp. Probasidia cylindrical to clavate, sub-hyaline, 30–47 × 4–9 μm, becoming bifurcate. Basidiospores reniform, with an apiculum at the base, thin- or thick-walled, pale yellow, 12.5–18 × 5–9.5 μm (av. 15 × 6.5 μm; n = 20), 1–3-septate, germination by means of conidial production and germ tubes.

Culture characteristics — Colonies of the primary mycelium attaining about 6 mm diam, velvety, orange to white. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6 × 3 μm.

Specimens examined. Japan, Nara, Kasuga Shrine, on dead branches of Podocarpus nagi, 23 Apr. 2006, T. Shirouzu, TNS-F-15721 (HNo.237); Mt Kasuga, on dead branches of an unknown broad-leaved tree, 23 Apr. 2006, T. Shirouzu, TNS-F-15720 (HNo.224), culture MAFF240135.

Notes — Dacrymyces minor is characterised by relatively small, pustulate to pulvinate basidiocarps, hyphae without clamp connections, and thin- or thick-walled, 1–3-septate basidiospores. This species was first recorded from Japan by Kobayasi (1939a).

(L.S. Olive) McNabb, New Zealand J. Bot. 11: 497. 1973 — Fig. 7h–m, 15b

Fig. 15

Basidiocarps. a. Dacrymyces minor TNS-F-15720 (HNo.224); b. Dacrymyces minutus TNS-F-15722 (HNo.282); c. Dacrymyces novae-zelandiae TNS-F-21038 (HNo.225); d. Dacrymyces pinacearum TNS-F-21056 (HNo.418); e. Dacrymyces punctiformis TNS-F-15723 (HNo.196); f. Dacrymyces san-augustinii TNS-F-21075 (HNo.666); g. Dacrymyces stillatus TNS-F-15727 (HNo.233); h. Dacrymyces subalpinus TNS-F-21071 (HNo.570); i. Dacrymyces subarcticus TNS-F-21067 (HNo.544); j. Dacrymyces unisporus TNS-F-15731 (HNo.332); k. Dacrymyces variisporus TNS-F-15733 (HNo.300); l. Dacryopinax spathularia TNS-F-15735 (HNo.379); m, n. Dacryopinax sphenocarpa TNS-F-21046 (HNo.356); o. Femsjonia peziziformis TNS-F-15737 (HNo439); p. Guepiniopsis buccina TNS-F-15738 (HNo.562). — Scale bars: a–p = 1 mm.

Basionym. Guepiniopsis minuta L.S. Olive, Bull. Torrey Bot. Club 81: 334. 1954.

Basidiocarps scattered, turbinate or irregularly discoid, sub-stipitate or stipitate bearing a centrally depressed semiglobose pileus, yellow-orange, firm-gelatinous, 1–3 mm high, 2–5 mm diam. Sterile parts of basidiocarps covered with simple cylindrical to clavate, hyaline, conspicuously thick-walled terminal cells, 25–32 × 6–10 μm. Internal hyphae branched, thin-walled, septate, hyaline, 2–3 μm diam, with clamp connections. Hymenium limited to the upper surface of the pileus or disc. Probasidia cylindrical to clavate, sub-hyaline to pale yellow, 30–38 × 5–7.5 μm, with a basal clamp connection, becoming bifurcate. Dikaryophyses cylindrical, simple, sub-hyaline, 45–52 × 2.5 μm, with a basal clamp connection. Basidiospores cylindrical to reniform, with an apiculum at the base, thin-walled, sub-hyaline to pale yellow, 11.5–15.5 × 4–6 μm (av. 14 × 5 μm; n = 20), 1–3-septate, germinated by production of conidia and germ tubes.

Culture characteristics — Colonies slow growing, attaining about 2 mm diam, velvety, orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Gunma, Mt Motoshirane, on dead branches of Abies veitchii, 28 May 2007, T. Shirouzu & D. Hirose, TNS-F-21073 (HNo.648); Nagano, Sugadairakougen, on dead branches of Larix leptolepis, 28 May 2006, T. Shirouzu, TNS-F-15722 (HNo.282), culture MAFF240137.

Notes — Dacrymyces minutus is characterised by turbinate to irregularly discoid basidiocarps, hyphae with clamp connections, cylindrical to clavate, thick-walled terminal cells, and thin-walled, 3-septate basidiospores. According to the results of our survey and that of McNabb (1973), D. minutus occurs frequently on woody materials of conifers. If D. pezizoides is thought of as a synonym of D. minutus as McNabb (1973) suggested, this species has already been reported from Japan by Kobayasi (1939a).

McNabb, New Zealand J. Bot. 11: 493. 1973 — Fig. 8a–e, 15c

Fig. 8

a–e. Dacrymyces novae-zelandiae TNS-F-21038 (HNo.225). a. Basi-diocarps; b. basidiospores; c. probasidium; d. developing basidium; e. dikaryophysis and probasidium. — f, g. Dacrymyces pinacearum TNS-F-21056 (HNo.418). f. Conidiocarps; g. conidia. — h–l. Dacrymyces punctiformis TNS-F-15723 (HNo.196). h. Basidiocarps; i. basidiospores; j. probasidium; k. developing basidium; l. dikaryophysis.

Basidiocarps scattered or gregarious, sometimes coalesced, pulvinate to convoluted, sessile or sub-stipitate, yellow, firm-gelatinous, 1–2 mm high, 1–4 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, sub-hyaline, 2–3 μm diam, without clamp connections. Hymenium limited to the upper surface of the basidiocarp. Probasidia cylindrical to clavate, thin-walled, pale yellow, 55–72 × 5–8.5 μm, becoming bifurcate. Dikaryophyses simple, thin-walled, pale yellow, 50–60 × 3–4 μm. Basidiospores cylindrical to reniform, with an apiculum at the base, thin-walled, yellow, 15–27 × 7–12.5 μm (av. 20.5 × 8.5 μm; n = 20), 0–7-septate, germination via the production of conidia and germ tubes.

Culture characteristics — Colonies attaining about 6 mm diam, velvety, white-orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6 × 3 μm.

Specimen examined. Japan, Nara, Kasuga Shrine, on dead branches of Podocarpus nagi, 23 Apr. 2006, T. Shirouzu, TNS-F-21038 (HNo.225), culture MAFF241165.

Notes — Dacrymyces novae-zelandiae is characterised by pustulate to pulvinate, sessile basidiocarps, hyphae without clamp connections, simple dikaryophyses and thin-walled, multi-septate basidiospores. This is the first record from Japan.

Shirouzu & Tokum., sp. nov. — MycoBank MB514038; Fig. 8f, g, 15d

Conidiomata sparsa, pulvinata vel flabellata, sessilia, aurantiaca, fragilia, 1 mm alta, 1–2 mm lata. Hyphae interaneae ramosae, tenuitunicatae, flavidae, 2–3 μm latae. Cellulae conidiogenae micronematicae, cylindricae, flavidae. Conidia holoblastica, ramosa, dendroidea, cum ramis cylindricis 1–8-cellularibus 27–65 × 5–9 μm tenuitunicatis aurantiacis.

Etymology. Named after the family name of its host.

Conidiocarps scattered, pulvinate to flabellate, sessile, orange, sometimes white at the lower-side, fragile, 1 mm high, 1–2 mm diam. Internal hyphae branched, thin-walled, septate, pale yellow, 2–3 μm diam, without clamp connections. Hymenium limited to the orange part of the basidiocarp. Conidiogenous cells micronematous, cylindrical, pale yellow. Conidia holoblastic, branched, dendroid, composed of cylindrical, 1–8-celled branches of 27–65 × 5–9 μm, thin-walled, orange.

Culture characteristics — Colonies attaining about 8 mm diam, velvety, white to pale yellow. Conidiogenesis not observed.

Specimen examined. Japan, Nara, Oodaigahara, on a dead trunk of Abies sp., 4 July 2006, T. Shirouzu, holotype TNS-F-21056 (HNo.418), culture ex-type MAFF241182.

Notes — Dacrymyces pinacearum is characterised by pulvinate to flabellate conidiocarps and holoblastic, dendroid conidia. Coelomycetous anamorphs of Dacrymycetales are usually linked with the genus Dacrymyces and have pulvinate fruit bodies and arthric conidia as represented by D. stillatus. An unusual coelomycetous anamorph of Dacrymycetales having holoblastic, 2–4-armed stauroconidia was described by Kirschner & Yang (2005) as Dacryoscyphus chrysochilus. Our fungus resembles the anamorphic species of Kirschner & Yang, but is different from D. chrysochilus in having dendroid conidia composed of branches without accurate tips. In molecular phylogenetic analyses, Dacrymyces pinacearum was clearly separate from Dacryoscyphus chrysochilus and formed a clade with Dacrymyces san-augustinii and D. novae-zelandiae (Fig. 16); therefore, we consider that this fungus is a new anamorphic species of Dacrymyces.

Neuhoff, Schweiz. Z. Pilzk. 12: 81. 1934 — Fig. 8h–l, 15e

For synonyms see Reid (1974).

Basidiocarps scattered or gregarious, sometimes coalesced, pustulate to pulvinate, sessile, pale yellow, firm-gelatinous, 0.5–1 mm high, 1–2 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, sub-hyaline, 2–3.5 μm diam, with clamp connections. Hymenium limited to the superior surface of the basidiocarp. Probasidia cylindrical to clavate, thick-walled, pale yellow, 30–50 × 5 μm, with a basal clamp connection, becoming bifurcate. Dikaryophyses simple, septate, thin-walled, sub-hyaline, 30–50 × 2.5 μm. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline, 7–13 × 4–6 μm (av. 9 × 5 μm; n = 15), 0–1(–3)-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 4 mm diam, velvety, white. Conidiogenesis not observed.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of Pinus densiflora, 20 Apr. 2006, T. Shirouzu, TNS-F-15723 (HNo.196), culture MAFF240138; Takaragaike, on dead branches of Clethra barbinervi, 21 Apr. 2006, T. Shirouzu, TNS-F-15724 (HNo.213); Nagano, Shioda, on dead branches of Pinus densiflora, 20 May 2006, T. Shirouzu, TNS-F-15725 (HNo.285).

Notes — Dacrymyces punctiformis is characterised by pustulate to pulvinate, sessile basidiocarps, hyphae with clamp connections, simple dikaryophyses and thin-walled, 0–1(–3)-septate basidiospores. This study as well as others (Kobayasi 1939a, McNabb 1973, Reid 1974), it was mentioned that this species frequently occurred on conifers wood.

Kobayasi, Sci. Rep. Tokyo Bunrika Daigaku, Sect. B, 4: 122. 1939 — Fig. 9a–e, 15f

Fig. 9

a–e. Dacrymyces san-augustinii TNS-F-21075 (HNo.666). a. Basi-diocarps; b. basidiospores; c. probasidium; d. developing basidium; e. dikaryophysis. — f–j. Dacrymyces stillatus TNS-F-15727 (HNo.233). f. Basidiocarps; g. basidiospores; h. probasidium; i. developing basidium; j. dikaryophysis.

Basidiocarps scattered or gregarious, sometimes coalesced, pustulate to pulvinate, sessile, pale orange to pale amber, firm-gelatinous, 1–2 mm high, 1–3 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, sub-hyaline, 2–3 μm diam, without clamp connections. Hymenium limited to the upper surface of the basidiocarp. Probasidia cylindrical to clavate, thin-walled, pale yellow, 38–58 × 5.5–7 μm, becoming bifurcate. Dikaryophyses simple or branched, septate, thin- or thick-walled, pale yellow, 40–120 × 4 μm. Basidiospores curved-allantoid, with an apiculum at the base, thin-walled, sub-hyaline, 16–27.5 × 6–10 μm (av. 21.5 × 7.5 μm; n = 20), 0–7-septate, germination via conidial production and germ tubes.

Culture characteristics — Colonies attaining about 20 mm diam, velvety, white-orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Okinawa, Iriomote Island, on dead branches of an unknown broad-leaved tree, 10 June 2007, T. Shirouzu, TNS-F-21075 (HNo.666), culture MAFF241196; Wakayama, Mt Shirami, on dead branches of an unknown woody plant, 8 July 2006, T. Shirouzu, TNS-F-15726 (HNo.441).

Notes — Dacrymyces san-augustinii is characterised by pustulate to pulvinate, sessile basidiocarps, hyphae without clamp connections, branched dikaryophyses and thin-walled, multi-septate basidiospores. Dacrymyces novae-zelandiae resembles D. san-augustinii in morphological characteristics. McNabb (1973) noted that these species can be distinguished by the presence of branched dikaryophyses, which are present in D. san-augustinii and absent in D. novae-zelandiae. This species was described by Kobayasi (1939a) on a specimen collected from the trunk of a broad-leaved tree in Japan.

Nees, Syst. Mycol. 2: 250. 1822 — Fig. 9f–j, 15g

For synonyms see McNabb (1973) and Reid (1974).

Basidiocarps scattered, pustulate, pulvinate to applanate, sometimes centrally depressed, sessile, pale yellow to pale amber, firm-gelatinous, 0.5–1 mm high, 1–3 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled marginal hyphae. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–3 μm diam, without clamp connections. Hymenium limited to the upper surface of the basidiocarp. Probasidia cylindrical to clavate, sub-hyaline, 25–42.5 × 4.5–7 μm, becoming bifurcate. Basidiospores reniform, with an apiculum at the base, thick-walled, pale yellow, 12.5–17 × 5–8 μm (av. 14.5 × 6 μm; n = 20), 1–3-septate, germination via the production of conidia and germ tubes. Anamorphic fruit bodies pustulate to pulvinate, sessile, orange, firm-gelatinous, 0.5 mm high, 0.5–1.5 mm diam. Conidia arthric, cylindrical, thin-walled, pale orange, 7.5–12.5 × 2.5–3.5 μm, 0–1-septate.

Culture characteristics — Colonies attaining about 15 mm diam, velvety, pale orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Kyoto, Midorogaike, on dead branches of Quercus glauca, 1 July 2006, T. Shirouzu, TNS-F-21052 (HNo.383); Nara, Kasuga Shrine, on dead branches of Podocarpus nagi, 23 Apr. 2006, T. Shirouzu, TNS-F-15727 (HNo.233), culture MAFF240142; Mt Tamaki, on dead branches of Deutzia crenata, 27 Apr. 2006, T. Shirouzu, TNS-F-15729 (HNo.252; anamorphic stage); on dead branches of an unknown conifer, 27 Apr. 2006, T. Shirouzu, TNS-F-15728 (HNo.256); Oodaigahara, on dead branches of Fagus crenata, 4 July 2006, T. Shirouzu, TNS-F-21057 (HNo.421; anamorphic stage); on dead branches of an unknown conifer, 4 July 2006, T. Shirouzu, TNS-F-21055 (HNo.411; anamorphic stage).

Notes — Dacrymyces stillatus is characterised by pustulate to pulvinate basidiocarps, hyphae without clamp connections and thick-walled, 1–3-septate basidiospores. This species is similar to D. minor in morphology on the whole, but McNabb (1973) emphasised that these species could be distinguished by the difference in basidiocarp size. In this study, we identified materials with basidiocarps of 2 mm or more in diameter as D. stillatus, but from the results of molecular phylogenetic analyses, these two species were nested in the same clade (Fig. 16). This species was reported in Japan by Kobayasi (1939a; as D. deliquescens) and Tubaki & Hosoya (1987).

Kobayasi, Sci. Rep. Tokyo Bunrika Daigaku, Sect. B, 4: 120. 1939 — Fig. 10a–e, 15h

Fig. 10

a–e. Dacrymyces subalpinus TNS-F-21071 (HNo.570). a. Basidiocarp; b. basidiospores; c. probasidium; d. basidium; e. marginal hyphae. — f, g. Dacrymyces subarcticus TNS-F-21067 (HNo.544). f. Conidiocarps; g. conidia.

Basidiocarps scattered or gregarious, sometimes coalesced, turbinate to cerebriform, sessile or stipitate, bearing a rugose to convoluted, semiglobose pileus, yellow to orange, firm-gelatinous, 1–2 mm high, 4–6 mm diam. Sterile parts of basidiocarps covered with simple, cylindrical to clavate, septate, 2–3-celled, hyaline, conspicuously thick-walled terminal cells, 38–50.5 × 7–12 μm. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–4 μm diam, without clamp connections. Hymenium limited to superior surface of the pileus. Probasidia cylindrical, yellow, 63.5–90 × 8–11.5 μm, becoming bifurcate. Basidiospores cylindrical to curved-naviculate, with an apiculum at the base, thin-walled, yellow, 32–54 × 9.5–17 μm (av. 41.5 × 12 μm; n = 20), 0–14-transverse septate, occasionally with 0–7-vertical septa, germinated by production of conidia and germ tubes.

Culture characteristics — Colonies attaining about 6 mm diam, velvety, pale yellow to white. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6–7 × 4 μm.

Specimens examined. Japan, Gifu, Mt Kiso-ontake, on dead branches of Tsuga sieboldii, 25 Oct. 2006, T. Osono, TNS-F-21071 (HNo.570), culture MAFF241193; Nara, Mt Kasuga, on dead branches of Abies firma, 23 Apr. 2006, T. Shirouzu, TNS-F-15730 (HNo.228).

Notes — Dacrymyces subalpinus is characterised by hyphae without clamp connections and curved-naviculate, multi-transverse and vertically septate basidiospores. Although McNabb (1973) noted that the holotype was destroyed during the Second World War, he recognised this as a distinct species based on the descriptions by Kobayasi (1939a).

Shirouzu & Tokum., sp. nov. — MycoBank MB514039; Fig. 10f, g, 15i

Conidiomata sparsa, papilliformia vel pulvinata, sessilia, flavida, fragilia, 0.5–1 mm alta, 1–2 mm lata. Hyphae interaneae ramosae, tenuitunicatae, flavidae, 2–3 μm latae. Cellulae conidiogenae micronematicae, cylindricae, flavidae. Conidia holoblastica, irregulariter ramosa, cum ramis cylindricis 1–4-cellularibus 6–22.5 × 2–4 μm tenuitunicatis flavidis.

Etymology. Named after its subarctic habitat.

Conidiocarps scattered, pustulate to pulvinate, sessile, yellow, fragile, 0.5–1 mm high, 1–2 mm diam. Internal hyphae branched, thin-walled, septate, pale yellow, 2–3 μm diam, without clamp connections. Hymenium limited to the upper surface of the basidiocarp. Conidiogenous cells micronematous, cylindrical, pale yellow. Conidia holoblastic, irregularly branched, composed of cylindrical, 1–4-celled branches of 6–22.5 × 2–4 μm, with a separation scar at the base, thin-walled, pale yellow.

Culture characteristics — Colonies attaining about 6 mm diam, velvety, white to pale yellow. Conidiogenesis not observed.

Specimens examined. Japan, Gunma, Mt Motoshirane, on a dead trunk of an unknown conifer, 23 July 2007, T. Shirouzu & D. Hirose, TNS-F-21076 (HNo.722); Nara, Oodaigahara, on dead branches of Abies sp., 9 Oct. 2006, T. Shirouzu, holotype TNS-F-21067 (HNo.544).

Notes — Dacrymyces subarcticus is characterised by pulvinate conidiocarps and holoblastic, irregularly branched conidia. This anamorphic fungus has pulvinate conidiocarps and is similar to sporodochial anamorphs, such as D. stillatus, in the shape of conidiocarps. However, D. subarcticus differs from other sporodochial anamorphs of Dacrymyces in bearing holoblastic, irregularly branched conidia. We described this fungus as a new anamorphic species of Dacrymyces. In molecular phylogenetic analyses, D. subarcticus formed a monophyletic group with Dacryoscyphus chrysochilus (Fig. 16).

(L.S. Olive) K. Wells, Mycologia 86: 31. 1994 — Fig. 11a–c, 15j

Fig. 11

a–c. Dacrymyces unisporus TNS-F-15731 (HNo.332). a. Basidiocarps; b. basidiospores; c. basidia and probasidium. — d–h. Dacrymyces variisporus TNS-F-15733 (HNo.300). d. Basidiocarps; e. basidiospores; f. probasidium and basidium; g. dikaryophysis; h. marginal hyphae.

Basionym. Platygloea unispora L.S. Olive, J. Elisha Mitchell Sci. Soc. 60: 17. 1944.

Basidiocarps scattered, pustulate or pulvinate, sometimes centrally depressed, sessile, olive-orange to yellow-orange, firm-gelatinous, 0.5 mm high, 0.5–1 mm diam. Sterile parts of basidiocarps covered with simple or branched, cylindrical, septate, hyaline, thin-walled, marginal hyphae. Internal hyphae branched, thick-walled, septate, hyaline, 3–5 μm diam, with clamp connections. Hymenium limited to the superior surface of the basidiocarp. Basidia cylindrical to clavate, tapering toward the apex bearing a sterigma, sub-hyaline, 40–58 × 4–7 μm, with a basal clamp connection. Basidiospores globose to subglobose, with an apiculum at the base, thin-walled, sub-hyaline, 11–17.5 × 8.5–13 μm (av. 14 × 11.5 μm; n = 20), 2–4-celled, germinated by germ tubes.

Culture characteristics — Colonies slow growing, attaining about 2 mm diam, lanose, white. Conidiogenesis not observed.

Specimen examined. Japan, Nagano, Sugadairakougen, on dead branches of Pinus densiflora, 16 June 2006, T. Shirouzu, TNS-F-15731 (HNo.332), culture MAFF240146.

Notes — The most remarkable morphological feature of Dacrymyces unisporus is a simple, not bifurcate, cylindrical basidium producing a single, globose basidiospore. This species was initially described as a species of Platygloea (Olive 1944). In 1994, Wells transferred this fungus to the genus Dacrymyces based on the fact that “very often the epibasidium and sterigma arise eccentirically suggesting a derivation from the furcated, dacrymycetoid basidium” (Wells 1994). This is the first record from Japan.

McNabb, New Zealand J. Bot. 11: 504. 1973 — Fig. 11d–h, 15k

Basidiocarps scattered or gregarious, sometimes coalesced, turbinate, pulvinate or discoid, centrally depressed, sessile or sub-stipitate, orange, firm-gelatinous, 0.5–1 mm high, 0.5–1.5 mm diam. Sterile parts of basidiocarps covered with cylindrical-clavate or narrowly naviculate, simple or branched, straight or flexuous, conspicuously thick-walled, hyaline terminal cells, 17.5–32 × 5–10 μm. Internal hyphae branched, thin- or thick-walled, septate, hyaline, 2–4.5 μm diam, with clamp connections. Hymenium limited to the superior surface of the basidiocarp. Probasidia cylindrical to clavate, yellow, 44.5–62.5 × 4.5–7.5 μm, with a basal clamp connection, becoming bifurcate. Dikaryophyses cylindrical, simple, septate, pale yellow, 30–50 × 3 μm. Basidiospores cylindrical to curved-cylindrical, with an apiculum at the base, thick-walled, yellow, 18–25 × 8–10.5 μm (av. 22 × 9 μm; n = 20), 1–7-transverse septate, occasionally with 1–4-vertical septa, germination via the production of conidia and germ tubes.

Culture characteristics — Colonies attaining about 4 mm diam, velvety, sulcate, yellow-orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of Pinus densiflora, 30 June 2006, T. Shirouzu, TNS-F-15734 (HNo.352); Nagano, Sugadairakougen, on dead branches of P. densiflora, 21 May 2006, T. Shirouzu, TNS-F-15733 (HNo.300), culture MAFF240148; Nara, Mt Tamaki, on dead branches of P. densiflora, 27 Apr. 2006, T. Shirouzu, TNS-F-15732 (HNo.263).

Notes — Dacrymyces variisporus is characterised by relatively small, turbinate to pulvinate basidiocarps, hyphae with clamp connections, cylindrical to clavate, thick-walled terminal cells, and thick-walled, multi-septate basidiospores. All our materials were collected from woody substrata of conifers. This is the first record from Japan.

(Schwein.) G.W. Martin, Lloydia 11: 116. 1948. — Fig. 12a–e, 15l

Fig. 12

a–e. Dacryopinax spathularia TNS-F-15735 (HNo.379). a. Basidiocarps; b. basidiospores; c. probasidium; d. basidium; e. marginal hyphae. — f–j. Dacryopinax sphenocarpa TNS-F-21046 (HNo.356). f. Basidiocarps; g. basidiospores; h. probasidium; i. basidium; j. marginal hyphae.

Basionym. Merulius spathularia Schwein., Schr. Naturf. Ges. Leipzig 1: 97. 1882.

For other synonyms see McNabb (1965b).

Basidiocarps scattered or gregarious, spathulate, stipitate bearing sinuate flabellate to petaloid pileus, orange, white-yellow at the sterile surface of basidiocarps, soft-cartilaginous, 5–13 mm high, 1–2 mm diam at stipe, 3–7 mm diam at pileus. Sterile parts of basidiocarps covered with solitary or fasciculate, cylindrical, simple or branched, straight or flexuous, septate, thick-walled, pale yellow marginal hypha of 30–140 × 5–6 μm. Internal hyphae branched, thin-walled, septate, pale yellow, 2–5 μm diam, without clamp connections. Hymenium unilateral. Probasidia cylindrical to clavate, pale yellow, 23.5–34 × 3.5–7.5 μm, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, pale yellow, 9–13.5 × 3.5–7.5 μm (av. 10.5 × 5 μm; n = 20), 0–1-septate, germinated by germ tubes.

Culture characteristics — Colonies attaining about 10 mm diam, velvety, yellow to white. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 5 × 3 μm.

Specimens examined. Japan, Kyoto, Midorogaike, on a bench made from an unknown conifer, 1 July 2006, T. Shirouzu, TNS-F-15735 (HNo.379), culture MAFF240150; Nara, Kasuga Shrine, on a handrail made from an unknown conifer, 3 July 2006, T. Shirouzu, TNS-F-15736 (HNo.398); Wakayama, Mt Nachi, on a fallen tree of an unknown palm, 6 July 2006, T. Shirouzu, TNS-F-21048 (HNo.367).

Notes — Dacryopinax spathularia is characterised by spathulate to stipitate basidiocarps with a flabellate pileus, thick-walled marginal hypha, and 0–1-septate basidiospores. This species has been reported from Japan by Kobayasi (1939b) as Guepinia spathularia.

Shirouzu & Tokum., sp. nov. — MycoBank MB514040; Fig. 12f–j, 15m, n

Basidiocarpia sparsa vel gregaria, spathulata vel subulata, albo-flavida vel sub-succinea, cartilaginea, 2–6 mm alta, 0.5–1 mm lata. Pili corticales steriles solitarii vel fasciculati, cylindrici vel clavati, tenuitunicati vel crassitunicati, sub-hyalini, 15–25 × 3.5–6 μm. Hyphae interaneae ramosae, tenuitunicatae, hyalinae, 2–4 μm latae, cum colligationibus unciformibus. Probasidia cylindrica vel clavata, hyalina, 15–34 × 3.5–8 μm, bifurcatascentia. Basidiosporae subglobosae vel reniformae, tenuitunicatae, hyalinae, 10–16 × 4.5–8.5 μm, 0–1-septatae.

Etymology. Named after its subulate basidiocarps.

Basidiocarps scattered or gregarious, spathulate to subulate, sometimes slightly branched at apex, yellow-white to pale amber, sometimes with simple root, soft-cartilaginous, 2–6 mm high, 0.5–1 mm diam. Sterile parts of basidiocarps covered with solitary or fasciculate, cylindrical to clavate, simple or branched, straight or flexuous, septate, thin- or thick-walled, sub-hyaline marginal hypha, 15–25 × 3.5–6 μm. Internal hyphae branched, thin-walled, septate, hyaline, 2–4 μm diam, with clamp connections. Hymenium unilateral, sometimes amphigenous. Probasidia cylindrical to clavate, hyaline, 15–34 × 3.5–8 μm, with a basal clamp connection, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, hyaline, 10–16 × 4.5–8.5 μm (av. 12.5 × 6 μm; n = 20), 0–1-septate, germination via germ tubes.

Culture characteristics — Colonies attaining about 6 mm diam, wet, yellow to orange. Conidiogenesis not observed.

Specimens examined. Japan, Kyoto, Mt Daimonji, on dead branches of an unknown conifer, 30 June 2006, T. Shirouzu, holotype TNS-F-21046 (HNo.356), culture ex-type MAFF241173; Mt Kiyomizu, on dead branches of Pinus densiflora, 7 Oct. 2006, T. Shirouzu, TNS-F-21066 (HNo.534); Nara, Mt Tamaki, on dead branches of P. densiflora, 7 July 2006, T. Shirouzu, TNS-F-21059 (HNo.430); Wakayama, Mt Nachi, on dead branches of Cryptomeria japonica, 6 July 2006, T. Shirouzu, TNS-F-21047 (HNo.364); 11 Oct. 2006, T. Shirouzu, TNS-F-21068 (HNo.552).

Notes — Dacryopinax sphenocarpa is characterised by sharp, spathulate basidiocarps, hyphae with clamp connections, solitary or fasciculate, thin- or thick-walled marginal hypha, and thin-walled, 0–1-septate basidiospores. This dacrymycetous fungus belongs to the genus Dacryopinax because of its basidiocarp morphology, having a unilateral hymenium and fasciculate, thick-walled marginal hypha. Only D. taibaishanensis is known as a species having hyphae with clamp connections in this genus (Liu & Fan 1990). Dacryopinax sphenocarpa is different from D. taibaishanensis in the shape of basidiocarps (D. taibaishanensis is discoid) and in the size (20–35 × 7–10 μm) and septal number (5–7-septate) of basidiospores. In molecular phylogenetic analyses, this species did not cluster in a monophyletic clade with another species of Dacryopinax, D. spathularia, but represented a monophyletic group with Dacrymyces ancyleus (Fig. 16).

(Lév.) P. Karst., Bidrag Kannedam Finlands Natur Folk 31: 352. 1876 — Fig. 13a–f, 15o

Fig. 13

a–f. Femsjonia peziziformis TNS-F-15737 (HNo.439). a. Basidiocarps; b. basidiospores; c. probasidium; d. developing basidium; e. dikaryophysis; f. marginal hyphae. — g–k. Guepiniopsis buccina TNS-F-15738 (HNo.562). g. Basidiocarps; h. basidiospores; i. probasidium; j. developing basidium; k. marginal hyphae.

Basionym. Exidia peziziformis Lév., Ann. Sci. Nat., Bot. 9: 127. 1848.

For other synonyms see McNabb (1965e).

Basidiocarps scattered or gregarious, turbinate to discoid, centrally depressed, sessile, yellow at hymenium, white rough tomentose at the sterile surface of basidiocarps, firm-gelatinous, 3–10 mm high, 5–10 mm diam. Sterile parts of basidiocarps densely covered with cylindrical to clavate, straight or flexuous, conspicuously thick-walled, hyaline to pale yellow terminal cells, 40–80 μm long, 4–7.5 μm diam. Internal hyphae branched, thin-walled, septate, hyaline, 2–5 μm diam, with clamp connections. Hymenium limited to the superior surface of the basidiocarp. Probasidia cylindrical to clavate, pale orange, 40–80 × 5.5–10.5 μm, with a basal clamp connection, becoming bifurcate. Dikaryophyses cylindrical, simple, septate, pale orange, 48–80 × 3–4 μm. Basidiospores cylindrical to curved-cylindrical, with an apiculum at the base, thin-walled, pale orange, 22–36 × 8.5–13 μm (av. 26.5 × 9.5 μm; n = 20), 1–13-septate, germinated by production of conidia and germ tubes.

Culture characteristics — Colonies attaining about 10 mm diam, wet, white-orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 6 × 3 μm.

Specimen examined. Japan, Nagano, Sugadairakougen, on dead branches of Pinus densiflora, 27 Sept. 2006, T. Shirouzu & T. Hosoya, TNS-F-15737 (HNo.439), culture MAFF240152.

Notes — Femsjonia peziziformis is characterised by turbinate to discoid basidiocarps that are tomentose in the sterile part, with cylindrical, thick-walled terminal cells, and multi-septate basidiospores. Reid (1974) treated this genus as a synonym of Ditiola peziziformis (as D. pezizaeformis). In this study, we classified this fungus in Femsjonia according to the opinion of McNabb (1965e). This species has been recorded from Japan by Kobayasi (1939b) as F. luteoalba, a synonym of F. peziziformis.

(Pers.) L.L. Kenn., Mycologia 50: 888. 1958 — Fig. 13g–k, 15p

Basionym. Peziza buccina Pers., Syn. Meth. Fung. 2: 659. 1801.

For additional synonyms see Reid (1974).

Basidiocarps scattered, stipitate bearing a cupulate pileus, pale orange, firm-gelatinous, 4–5 mm high, 1.5–3 mm diam at the stipe, 2–3 mm high, 3–4 mm diam at the pileus. Sterile parts of basidiocarps densely covered with simple, cylindrical to clavate, septate, 2–3-celled, hyaline, conspicuously thick-walled terminal cells, 35–50 × 8–14.5 μm. Internal hyphae branched, thin-walled, gelatinous, septate, hyaline, 2–5 μm diam, without clamp connections. Hymenium limited in the cup of the pileus. Probasidia cylindrical, sub-hyaline, 31.5–55 × 3.5–8 μm, becoming bifurcate. Basidiospores subglobose to reniform, with an apiculum at the base, thin-walled, sub-hyaline, 10–16 × 5–7 μm (av. 13 × 6 μm; n = 10), 0–3-septate, germinated by germ tubes.

Culture characteristics — Colonies attaining about 5 mm diam, velvety, yellow to orange. Conidiogenous cells on vegetative hyphae, polyblastic, sympodial. Conidia subglobose, 4 × 3 μm.

Specimen examined. Japan, Nara, Mt Tamaki, on dead branches of an unknown broad-leaved tree, 13 Oct. 2006, T. Shirouzu, TNS-F-15738 (HNo.562), culture MAFF240153.

Notes — Guepiniopsis buccina is characterised by stipitate basidiocarps with cupulate pilei, thick-walled terminal cells, and 0–3-septate basidiospores. This species has been recorded from Japan as G. melurinus (Kobayasi 1939a).

Molecular phylogenetic analysis

The molecular phylogenetic tree using D1/D2 sequences is shown in Fig. 16. Four clades in Fig. 16 were defined as Dacrymycetes, Dacrymycetales, Cerinomycetaceae and Dacrymycetaceae, of which the Dacrymycetes, Dacrymycetales and Cerinomycetaceae clades were well supported (BS value (MP/ML) = 84/95 %, 90/93 %, 100/100 %, respectively). However, the Dacrymycetaceae clade (BS value (MP/ML) = 68/55 %) and other relatively higher-level nodes, were not well supported. At the species level, several clades such as the Guepiniopsis buccina, Calocera viscosa and Dacrymyces punctiformis clades were well supported (BS values = 100 %).

Species of the genus Dacrymyces were dispersed widely over the Dacrymycetes lineage. Dacrymyces stillatus and D. minor formed a clade with Guepiniopsis buccina (BS value (MP/ML) = 75/64 %), and Dacrymyces punctiformis formed a well-supported monophyletic group with Cerinomyces species (BS value (MP/ML) = 100/100 %).

Dacrymyces stillatus and D. minor were paraphyletic. Morphologically similar species, D. aureosporus and D. chrysospermus, represented a monophyletic group, but they were distributed in different sub-clades (BS value (MP/ML) = 74/69 %, 62/<50 %, respectively). Dacrymyces unisporus was located in the Dacrymycetes clade. Dacrymyces subarcticus formed a monophyletic group with Dacryoschyphus chrysochilus (BS value (MP/ML) = 100/100 %). Two Dacryopinax species, D. shenocarpus and D. spathularia, did not form a monophyletic group.

DISCUSSION

Japanese Dacrymycetes

In this study we recognised 28 species, including 5 new species and 9 species that were newly recorded from Japan (Table 3). Based on our survey, the total number of dacrymycetous species recorded from Japan increased from 28 to 42, although we were unable to recollect all species recorded by Kobayasi (1939a, b). This result suggests that the species diversity of dacrymycetous fungi in Japan is richer than previously thought.

The basidiocarps of Dacrymycetes could be widely collected throughout the Japanese Islands, from Hokkaido (subarctic coniferous forests) to the Yaeyama Islands (subtropical evergreen forests). However, dacrymycetous species and samples were more commonly collected from coniferous and deciduous broad-leaved forests in the subarctic and cool-temperate zone, than in the evergreen, broad-leaved forests in the warm-temperate and subtropical zone. The main diversity of Dacrymycetes may thus be from the cool-temperate to the subarctic zone.

To date, anamorphic stages with conidiocarps are known in Dacrymyces lacrymalis (present study), D. capitatus, D. minor and D. stillatus, (Oberwinkler 1993). A unique coelomycetous species, Dacryoscyphus chrysochilus (Kirschner & Yang 2005), and a hyphomycetous species, Cerinosterus luteoalbus, have also been reported (Moore 1987, Middelhoven et al. 2000). Among the newly described Dacrymyces species in this study, D. pianacearum and D. subarcticus were anamorphic species. In their fruit bodies (conidiocarps), only conidia were observed, and their holomorphs remain to be discovered. They also had various shapes of conidiocarps (flabellate and pustulate to pulvinate) and conidia (dendroid and irregularly branched). These new Japanese coelomycetous species suggest an unexpected high diversity of anamorphic stages in Dacrymycetes.

Some specimens agreed with the original description of D. subalpinus (Kobayasi 1939a). It had been thought that the holotype of this species was destroyed in the past (McNabb 1973). In this study, we reconfirmed the existence of D. subalpinus, and were able to recollected and deposit fresh specimens; however, many holotypes of other Japanese dacrymycetous species described by Kobayasi (1939a, b) have still not been found. During this study, specimens corresponding to such problematic species could not be recollected and examined (Table 1, 3), and hence their taxonomical validity remains uncertain. To confirm these doubtful species, future investigations of unlabelled dacrymycetous specimens deposited in TNS may provide useful additional information. It is suspected that these specimens may include the lost holotypes of Dacrymycetes species previously described from Japan.

One higher node in the molecular phylogenetic tree, the Dacrymycetaceae clade, was not well supported, and other higher phylogenetic relationships in this clade could not be resolved (Fig. 16). To define these unclear phylogenetic relationships, it may be necessary to perform additional analyses using more dacrymycetous DNA sequences. Phylogenetic analyses using multi-gene sequences, which succeeded in revealing more detailed relationships in fungal lineages (e.g. James et al. 2006), may resolve the higher phylogenetic relationships in Dacrymycetes.

A cushion-shaped Dacrymycetaceae species, Dacrymyces punctiformis, was included in the Cerinomycetaceae clade (Fig. 16). In addition, an exceptional species having non-bifurcate basidia, D. unisporus, was located basally in the Dacrymycetales clade (Fig. 16). These results suggest that a reconsideration of not only the taxonomical circumscriptions of genera, but also that of families and orders is required in this class. To solve these taxonomic problems, we may have to search for other informative criteria that correlate with the higher classification.

The molecular phylogenetic analysis of the present study in most cases supported the validity of the species identifications (Fig. 16), and also revealed the polyphyletic arrangement of Dacrymyces and Dacryopinax species throughout the Dacrymycetes lineage (Fig. 16). The classification of dacrymycetous genera is mainly based on the external morphology of the basidiocarp, and the wall thickness of marginal hyphae (McNabb & Talbot 1973), but our results underline the difficulty of using these criteria as primary characters for generic classification. Taxonomic rearrangement of dacrymycetous genera is thus necessary (Shirouzu et al. 2007). At this time, however, we have no phenotypic clues to rearrange dacrymycetous genera; therefore, it is appropriate that the newly described species such as Dacrymyces ancyleus and Dacryopinax sphenocarpa belong to genera ‘Dacrymyces’ and ‘Dacryopinax’ according to the existing taxonomic framework. Presently we are working on phylogenetic analyses that incorporate more taxa and DNA sequences, with the eventual aim to present a new interpretation to the remaining taxonomic problems.