Revision of the Massarineae (Pleosporales, Dothideomycetes)

Abstract

We here taxonomically revise the suborder Massarineae (Pleosporales, Dothideomycetes, Ascomycota). Sequences of SSU and LSU nrDNA and the translation elongation factor 1-alpha gene (tef1) are newly obtained from 106 Massarineae taxa that are phylogenetically analysed along with published sequences of 131 taxa in this suborder retrieved from GenBank. We recognise 12 families and five unknown lineages in the Massarineae. Among the nine families previously known, the monophyletic status of the Dictyosporiaceae, Didymosphaeriaceae, Latoruaceae, Macrodiplodiopsidaceae, Massarinaceae, Morosphaeriaceae, and Trematosphaeriaceae was strongly supported with bootstrap support values above 96 %, while the clades of the Bambusicolaceae and the Lentitheciaceae are moderately supported. Two new families, Parabambusicolaceae and Sulcatisporaceae, are proposed. The Parabambusicolaceae is erected to accommodate Aquastroma and Parabambusicola genera nova, as well as two unnamed Monodictys species. The Parabambusicolaceae is characterised by depressed globose to hemispherical ascomata with or without surrounding stromatic tissue, and multi-septate, clavate to fusiform, hyaline ascospores. The Sulcatisporaceae is established for Magnicamarosporium and Sulcatispora genera nova and Neobambusicola. The Sulcatisporaceae is characterised by subglobose ascomata with a short ostiolar neck, trabeculate pseudoparaphyses, clavate asci, broadly fusiform ascospores, and ellipsoid to subglobose conidia with or without striate ornamentation. The genus Periconia and its relatives are segregated from the Massarinaceae and placed in a resurrected family, the Periconiaceae. We have summarised the morphological and ecological features, and clarified the accepted members of each family. Ten new genera, 22 new species, and seven new combinations are described and illustrated. The complete ITS sequences of nrDNA are also provided for all new taxa for use as barcode markers.

Introduction

The Pleosporales is the largest order in the Dothideomycetes. Two suborders have been recognised, viz. the Pleosporineae and the Massarineae, containing more than 19 families that presently still lack subordinal assignment (Hyde et al. 2013). One suborder, the Pleosporineae, includes many economically important plant pathogens, Alternaria, Bipolaris, Didymella, Leptosphaeria, Parastagonospora, and Pyrenophora, for example, containing huge species diversity (Manamgoda et al., 2011, Zhang et al., 2012, Woudenberg et al., 2013). In this suborder, the phylogenetic relationships within each family, the morphological circumscriptions including those of asexual morphs, their evolutionary trend as plant pathogens, biogeography, and speciation are well established and documented (Rouxel and Balesdent, 2005, Peever, 2007, Zhang et al., 2009b, Ohm et al., 2012, Grandaubert et al., 2014). In contrast, for the other suborder, the Massarineae, much fundamental information including the taxonomic framework, phylogenetic relationships, biology, and species diversity are poorly understood. The Massarineae was originally established by Barr (1979) to accommodate the Massarinaceae and the Arthopyreniaceae, but this suborder has long been disregarded in the classification of bitunicate ascomycetes (Hawksworth et al., 1983, Hawksworth et al., 1995, Barr, 1987, Eriksson and Winka, 1998, Kirk et al., 2008, Lumbsch and Huhndorf, 2010).

The family Massarinaceae was established by Munk (1956) to encompass four genera, Massarina, Metasphaeria, Pseudotrichia, and Trichometasphaeria. The thin basal peridium and dark tissue (= clypeus) around the ostiole found in the ascomata of these genera were emphasised as familial characters. Although this proposal was accepted by many researchers (Eriksson, 1981, Boise, 1985), the characters emphasised by Munk (1956) for the family also exist in several other groups of bitunicate ascomycetes, such as Didymosphaeria (Didymosphaeriaceae), Salsuginea (Salsugineaceae) and Roussoella (Roussoellaceae). The Massarinaceae, therefore, has been treated as a synonym of the Pleosporaceae (Bose, 1961, Luttrell, 1973, Von Arx and Müller, 1975) or Lophiostomataceae (Barr, 1987, Barr, 1992). Similarly, taxonomic circumscription of the genus Massarina has also been repeatedly revised (see Notes in Massarina), and as a consequence of a clear taxonomic definition of Massarina, the family Massarinaceae has been regarded as a natural lineage. Several related genera, e.g., Aquaticheirospora, Dictyosporium, and Periconia (Tsui et al., 2006, Kodsueb et al., 2007, Schoch et al., 2009), have been assigned to the Massarinaceae as circumscribed by Eriksson & Hawksworth (2003), and the concept of the family has been revised (Hyde et al. 2013).

In a recent molecular study on the Pleosporales (Zhang et al. 2012), the suborder Massarineae was resurrected as a sister to the Pleosporineae, and was emended to include five families, the Lentitheciaceae (Zhang et al. 2009b), Massarinaceae (Munk 1956), Montagnulaceae (Barr 2001; later synonymised under Didymosphaeriaceae, Ariyawansa et al. 2014), Morosphaeriaceae (Suetrong et al. 2009), and Trematosphaeriaceae (Suetrong et al. 2011b). More recently, the Bambusicolaceae (Hyde et al. 2013), Dictyosporiaceae (nom. prov., see Liu et al. 2015), Latoruaceae and Macrodiplodiopsidaceae (Crous et al. 2015a) have been added to the suborder. Most of these families have been recognised from the results of recent molecular studies. Only a few members of each family are currently known, and thus the morphological characteristics and phylogenetic relationships within each family are not fully understood. Consequently, several new genera belonging to the Massarineae, such as Ascorhombispora (Cai & Hyde 2007b), Inflatispora (Zhang et al. 2011), and Noosia (Crous et al. 2011a), have been published but their phylogenetic placements at familial level remain obscure (Zhang et al., 2012, Hyde et al., 2013).

Our aims were to reveal the species diversity within the Massarineae and to establish a taxonomic framework within this suborder for understanding the relationships among the Massarineae. To this end we examined 106 specimens/isolates belonging to this suborder, and analysed their morphology and partial DNA sequences of the small and large subunit nuclear ribosomal DNA (SSU and LSU nrDNA) and the translation elongation factor 1-alpha gene (tef1).

Materials and methods

Morphological studies

Leaf and twig specimens were collected from various plants in Japan, and deposited in the herbarium of Hirosaki University (HHUF). Measurements of all structures except for ascomata/conidiomata were taken from material mounted in distilled water. India ink or Black-Blue ink in distilled water was added to water mounts to detect gelatinous sheaths or appendages around spores. To observe the internal structure of strongly melanised spores, 5 % sodium hypochlorite solution (NaClO) was used for the bleaching of spores as described in Eriksson (1989). The position of the primary septum of spores was noted using the decimal system (Shoemaker 1984), and the numbers of spore septa were recorded as “septa of upper hemisphere + the primary septum + septa of lower hemisphere”. To observe sporocarp structure, ascomata/conidiomata were boiled in water for a few minutes, sectioned using a freezing microtome (HM 400R; MICROM, Germany), and mounted in diluted lactophenol cotton blue. Morphology was observed using differential interference and phase contrast microscopy (Olympus BX53, Japan).

Single spore cultures were obtained following the methods of Tubaki (1978). Ninety-nine cultures on 2 % potato-dextrose agar (PDA) or potato-carrot agar (PCA) were prepared from the collections and were deposited in the Japan Collection of Microorganisms (JCM), the National Institute of Agrobiological Sciences, Japan (MAFF), and the CBS-KNAW Fungal Biodiversity Centre (Centraalbureau voor Schimmelcultures; CBS). An additional seven strains were obtained from the CBS culture collection (Table 1). Growth rate and colony characteristics were recorded from cultures grown on PDA within 2 or 4 wk at 20 °C in the dark. Colours were designated according to Rayner (1970). Induction of sexual/asexual sporulation was attempted by culturing isolates on rice straw agar (RSA; Tanaka & Harada 2003a) and/or incubating small pieces of colony in sterilised water (Scheuer 1991). Nomenclatural novelties were deposited in MycoBank (Crous et al. 2004).

Table 1.

Cultures and GenBank accession numbers of Massarineae obtained in this study.

Taxon	Family1	Original no.	Culture no.	Specimen no.2	GenBank accession no.3				Notes4
SSU	LSU	tef1	ITS
Aquastroma magniostiolata	Par	KT 2485	CBS 139680 = JCM 19429 = MAFF 243824	HHUF 30122HT	AB797220	AB807510	AB808486	LC014540	A
Aquilomyces rebunensis	Mor	KT 732-2	CBS 139684 = JCM 19427 = MAFF 243862	HHUF 27556HT	AB797252	AB807542	AB808518	AB809630	A
Bactrodesmium cubense	IS	–	CBS 680.96 = JCM 14126	–	AB797218	AB807508	AB808484	LC014541	C
Clypeoloculus akitaensis	Mor	KT 788	CBS 139681 = JCM 19424 = MAFF 239467	HHUF 27557HT	AB797253	AB807543	AB808519	AB809631	A
C. hirosakiensis	Mor	KT 1283	CBS 139682 = JCM 19425 = MAFF 243864	HHUF 30144HT	AB797260	AB807550	AB808526	AB809638	A
C. microsporus	Mor	KT 1131	CBS 139683 = JCM 19426 = MAFF 243863	HHUF 30143HT	AB797245	AB807535	AB808510	AB811451	A
C. towadaensis	Mor	KT 1340	CBS 139685 = JCM 19428 = MAFF 243865	HHUF 30145HT	AB797259	AB807549	AB808525	AB809637	A
Dictyosporium aff. bulbosum	Dic	KH 375	JCM 19403 = MAFF 243829	HHUF 30127	AB797224	AB807514	AB808490	LC014542	C
D. aff. heptasporum	Dic	KH 332	JCM 19406 = MAFF 243828	HHUF 30126	AB797223	AB807513	AB808489	LC014543	C
D. bulbosum	Dic	yone 221	MAFF 243835	HHUF 29990	AB797221	AB807511	AB808487	LC014544	C
D. digitatum	Dic	KH 401	JCM 19404 = MAFF 243830	HHUF 30128	AB797225	AB807515	AB808491	LC014545	C
D. digitatum	Dic	KT 2660	JCM 19405 = MAFF 243833	HHUF 30131	AB797228	AB807518	AB808494	LC014546	C
D. digitatum	Dic	yone 280	MAFF 243837	HHUF 30093	AB797222	AB807512	AB808488	LC014547	C
D. hughesii	Dic	KT 1847	JCM 19407 = MAFF 243832	HHUF 30130	AB797227	AB807517	AB808493	LC014548	C
D. pseudomusae	Dic	KH 412	JCM 19408 = MAFF 243831	HHUF 30129PT	AB797226	AB807516	AB808492	LC014549	C
D. pseudomusae	Dic	yone 234	CBS 139686 = JCM 19409 = MAFF 243836	HHUF 30133HT	AB797230	AB807520	AB808496	LC014550	C
D. tetrasporum	Dic	KT 2865	JCM 19410 = MAFF 243834	HHUF 30132	AB797229	AB807519	AB808495	LC014551	C
Fuscostagonospora sasae	IS	KT 1467	CBS 139687 = JCM 13104 = MAFF 239614	HHUF 29106HT	AB797258	AB807548	AB808524	AB809636	A
Gregarithecium curvisporum	Dic	KT 922	CBS 139688 = JCM 19411 = MAFF 243838	HHUF 30134HT	AB797257	AB807547	AB808523	AB809644	A
Helicascus aquaticus	Mor	KT 1544	JCM 19423 = MAFF 243866	HHUF 30146	AB797242	AB807532	AB808507	AB809627	A
H. elaterascus	Mor	KT 2673	MAFF 243867	HHUF 30147	AB797243	AB807533	AB808508	AB809626	A
H. elaterascus	Mor	KT 2682	CBS 139689	HHUF 30451	LC014603	LC014608	LC014613	LC014552	A
H. thalassioideus	Mor	–	CBS 110441 = JCM 14147	–	AB797267	AB807557	AB808533	LC014553	A
H. thalassioideus	Mor	KH 242	JCM 17526 = NBRC 107811	HHUF 30069	AB797268	AB807558	AB808534	LC014554	A
Helminthosporium dalbergiae	Mas	H 4628 (= TS 36)	MAFF 243853	HHUF 27971	AB797231	AB807521	AB808497	LC014555	C
H. magnisporum	Mas	H 4627 (= TS 33)	MAFF 239278	HHUF 27968HT	AB797232	AB807522	AB808498	AB811452	C
H. massarinum	Mas	KT 838	JCM 13094 = MAFF 239604	HHUF 27573PT	AB797233	AB807523	AB808499	AB809628	A
H. massarinum	Mas	KT 1564	CBS 139690 = JCM 13095 = MAFF 239605	HHUF 29089HT	AB797234	AB807524	AB808500	AB809629	A
Helminthosporium sp.	Mas	H 4743 (= TS 68)	MAFF 243856	HHUF 28248	AB797236	AB807526	–	–	C
Helminthosporium sp.	Mas	yone 38	MAFF 243857	HHUF 29740	AB797237	AB807527	AB808502	–	C
Helminthosporium sp.	Mas	yone 63	MAFF 243858	HHUF 29741	AB797238	AB807528	AB808503	–	C
H. velutinum	Mas	H 4626 (= TS 28)	MAFF 243854	HHUF 27966	AB797240	AB807530	AB808505	LC014556	C
H. velutinum	Mas	H 4739 (= TS 58)	MAFF 243855	HHUF 28243	AB797235	AB807525	AB808501	LC014557	C
H. velutinum	Mas	yone 96	MAFF 243859	HHUF 30140	AB797239	AB807529	AB808504	LC014558	C
Karstenula rhodostoma	Did	–	CBS 691.94	UPS (F-141152) 425947	AB797241	AB807531	AB808506	LC014559	A
Katumotoa bambusicola	Len	KT 1517a	JCM 13131 = MAFF 239641	HHUF 28661PT	AB5244541	AB5245951	AB5391082	LC014560	A
Keissleriella breviasca	Len	KT 540	JCM 19413 = MAFF 239476	HHUF 27715PT	AB797296	AB807586	AB808565	AB811453	A
K. breviasca	Len	KT 581	JCM 19414 = MAFF 243843	HHUF 27717PT	AB797297	AB807587	AB808566	AB811454	A
K. breviasca	Len	KT 649	CBS 139691 = JCM 19415 = MAFF 243844	HHUF 27718HT	AB797298	AB807588	AB808567	AB811455	A
K. culmifida	Len	KT 2308	JCM 19416 = MAFF 243848	HHUF 30135	AB797301	AB807591	AB808570	LC014561	A
K. culmifida	Len	KT 2642	JCM 19417 = MAFF 243849	HHUF 30136	AB797302	AB807592	AB808571	LC014562	A
K. gloeospora	Len	KT 829	MAFF 239474	HHUF 27704	AB797299	AB807589	AB808568	LC014563	A
K. quadriseptata	Len	KT 2292	CBS 139692 = JCM 19418 = MAFF 243850	HHUF 30137HT	AB797303	AB807593	AB808572	AB811456	A
Keissleriella sp.	Len	KT 895	JCM 19420 = MAFF 243845	HHUF 27705	AB797300	AB807590	AB808569	–	A
K. taminensis	Len	KT 571	MAFF 243846	HHUF 27707	AB797305	AB807595	AB808574	LC014564	A
K. taminensis	Len	KT 594	MAFF 243847	HHUF 27709	AB797306	AB807596	–	–	A
K. taminensis	Len	KT 678	MAFF 239475	HHUF 27711	AB797307	AB807597	AB808575	LC014565	A
K. yonaguniensis	Len	KT 2604	CBS 139693 = JCM 19419 = MAFF 243851	HHUF 30138HT	AB797304	AB807594	AB808573	AB811457	A
Lentithecium clioninum	Len	KT 1149A	CBS 139694 = JCM 12703 = MAFF 239293	HHUF 28199HT	AB797250	AB807540	AB808515	LC014566	A
L. clioninum	Len	KT 1220	MAFF 243839	HHUF 28213PT	AB797251	AB807541	AB808516	LC014567	A
L. pseudoclioninum	Len	KT 1111	JCM 19421 = MAFF 243840	HHUF 29053PT	AB797254	AB807544	AB808520	AB809632	A
L. pseudoclioninum	Len	KT 1113	CBS 139695 = JCM 19422 = MAFF 243841	HHUF 29055HT	AB797255	AB807545	AB808521	AB809633	A
Magnicamarosporium iriomotense	Sul	KT 2822	CBS 139696 = JCM 19402 = MAFF 243827	HHUF 30125HT	AB797219	AB807509	AB808485	AB809640	C
Massarina cisti	Mas	–	CBS 266.62 = JCM 14140	ZT (Hütter & Loeffler)HT	AB797249	AB807539	AB808514	LC014568	A
M. eburnea	Mas	H 3953	CBS 139697 = JCM 14422	HHUF 26621	AB5217183	AB5217353	AB808517	LC014569	A
Monodictys capensis	IS	HR 1	CBS 134928 = VKM F-4506	HHUF 29712	AB797261	AB807551	AB808527	LC014570	C
Monodictys sp.	Par	JO 10	MAFF 243825	HHUF 30123	AB797262	AB807552	AB808528	–	C
Monodictys sp.	Par	KH 331	MAFF 243826	HHUF 30124	AB797263	AB807553	AB808529	–	C
Morosphaeria ramunculicola	Mor	KH 220	NBRC 107813	HHUF 30070	AB797264	AB807554	AB808530	–	A
M. velatispora	Mor	KH 218	JCM 17529 = NBRC 107814	HHUF 30072	AB797265	AB807555	AB808531	LC014571	A
M. velatispora	Mor	KH 221	JCM 17530 = NBRC 107812	HHUF 30073	AB797266	AB807556	AB808532	LC014572	A
Neokalmusia brevispora	Did	KT 1466	CBS 120248 = JCM 13543 = MAFF 239276	HHUF 28229	AB5244591	AB5246001	AB5391121	LC014573	A
N. brevispora	Did	KT 2313	NBRC 106240	HHUF 30016ET	AB5244601	AB5246011	AB5391132	LC014574	A
N. scabrispora	Did	KT 1023	CBS 120246 = JCM 12851 = MAFF 239517	HHUF 28608	AB5244521	AB5245931	AB5391062	LC014575	A
N. scabrispora	Did	KT 2202	NBRC 106237	HHUF 30013	AB5244531	AB5245941	AB5391072	LC014576	A
Neoophiosphaerella sasicola	Len	KT 1706	CBS 120247 = JCM 13134 = MAFF 239644	HHUF 29443ET	AB5244581	AB5245991	AB5391112	LC014577	A
Parabambusicola bambusina	Par	H 4321	MAFF 239462	HHUF 26590	AB797246	AB807536	AB808511	LC014578	A
P. bambusina	Par	KH 139	MAFF 243823	HHUF 30121	AB797247	AB807537	AB808512	LC014579	A
P. bambusina	Par	KT 2637	MAFF 243822	HHUF 30120	AB797248	AB807538	AB808513	LC014580	A
Paraphaeosphaeria michotii	Did	KT 2222	MAFF 243861	HHUF 30142	AB797269	AB807559	AB808535	AB809639	A
Periconia byssoides	Per	H 4600 (= TS 29)	MAFF 243872	HHUF 28238	AB797280	AB807570	AB808546	LC014581	C
P. byssoides	Per	H 4432	MAFF 243869	–	AB797279	AB807569	AB808545	LC014582	C
P. byssoides	Per	H 4853 (= TS 60)	MAFF 243873	–	AB797281	AB807571	AB808547	LC014583	C
P. digitata	Per	–	CBS 510.77	–	AB797271	AB807561	AB808537	LC014584	C
P. homothallica	Per	KT 916	CBS 139698 = JCM 13100 = MAFF 239610	HHUF 29105HT	AB797275	AB807565	AB808541	AB809645	A
P. igniaria	Per	–	CBS 379.86	–	AB797276	AB807566	AB808542	LC014585	C
P. igniaria	Per	–	CBS 845.96 = JCM 14142	–	AB797277	AB807567	AB808543	LC014586	C
P. pseudobyssoides	Per	H 4151	MAFF 243868	–	AB797278	AB807568	AB808544	LC014587	C
P. pseudobyssoides	Per	H 4790 ( = TS 102)	MAFF 243874	HHUF 28257	AB797270	AB807560	AB808536	LC014588	C
P. pseudodigitata	Per	KT 644	JCM 13164 = MAFF 239674	HHUF 27569PT	AB797272	AB807562	AB808538	LC014589	A
P. pseudodigitata	Per	KT 1195A	JCM 13165 = MAFF 239675	HHUF 29368PT	AB797273	AB807563	AB808539	LC014590	A
P. pseudodigitata	Per	KT 1395	CBS 139699 = JCM 13166 = MAFF 239676	HHUF 29370HT	AB797274	AB807564	AB808540	LC014591	A
Periconia sp.	Per	KT 1820A	MAFF 243870	HHUF 30148	AB797282	AB807572	AB808548	–	C
Periconia sp.	Per	KT 1825	MAFF 243871	HHUF 30149	AB797283	AB807573	AB808549	–	C
Pseudocoleophoma calamagrostidis	Dic	KT 3284	CBS 139700	HHUF 30450HT	LC014604	LC014609	LC014614	LC014592	A
P. polygonicola	Dic	KT 731	CBS 139701 = JCM 19412 = MAFF 239468	HHUF 27558HT	AB797256	AB807546	AB808522	AB809634	A
Setoseptoria arundinacea	Len	KT 552	MAFF 239460	HHUF 27543	AB797284	AB807574	AB808550	LC014594	A
S. arundinacea	Len	KT 600	MAFF 243842	HHUF 27544	AB797285	AB807575	AB808551	LC014595	A
S. magniarundinacea	Len	KT 1174	CBS 139702 = MAFF 239294	HHUF 28293HT	AB797286	AB807576	AB808552	LC014596	A
Spegazzinia deightonii	Did	yone 66	MAFF 243876	HHUF 30150	AB797291	AB807581	AB808557	–	C
S. deightonii	Did	yone 212	MAFF 243877	HHUF 30151	AB797292	AB807582	AB808558	–	C
Spegazzinia sp.	Did	yone 279	MAFF 243878	HHUF 30152	AB797293	AB807583	AB808559	–	C
S. tessarthra	Did	SH 287	MAFF 243875	HHUF 27691	AB797294	AB807584	AB808560	–	C
Stagonospora perfecta	Mas	KT 1726A	JCM 13099 = MAFF 239609	HHUF 29095	AB797289	AB807579	AB808555	AB809642	A
S. pseudoperfecta	Mas	KT 889	CBS 120236 = JCM 13097 = MAFF 239607	HHUF 29087HT	AB797287	AB807577	AB808553	AB809641	A
Stagonospora sp.	Mas	KT 903	CBS 120237 = JCM 13098 = MAFF 239608	HHUF 29088	AB797288	AB807578	AB808554	–	A
S. tainanensis	Mas	KT 1866	MAFF 243860	HHUF 30141	AB797290	AB807580	AB808556	AB809643	A
Sulcatispora acerina	Sul	KT 2982	CBS 139703	HHUF 30449HT	LC014605	LC014610	LC014615	LC014597	A
S. berchemiae	Sul	KT 1607	CBS 139704 = JCM 13101 = MAFF 239611	HHUF 29097HT	AB797244	AB807534	AB808509	AB809635	A
Tingoldiago graminicola	Len	KH 68	JCM 16485 = NBRC 106131	HHUF 30009HT	AB5217263	AB5217433	AB808561	LC014598	A
T. graminicola	Len	KH 155	JCM 16486 = NBRC 106132	HHUF 30010PT	AB5217283	AB5217453	AB808562	LC014599	A
T. graminicola	Len	KT 891	MAFF 239472	HHUF 27882PT	AB5217273	AB5217443	AB808563	LC014600	A
Trematosphaeria pertusa	Tre	KT 1496	JCM 19430 = MAFF 243879	HHUF 30153	AB797295	AB807585	AB808564	AB809646	A
T. pertusa	Tre	KT 3314	CBS 139705	HHUF 30452	LC014606	LC014611	LC014616	LC014601	A
T. pertusa	Tre	KT 3315	CBS 139706	HHUF 30453	LC014607	LC014612	LC014617	LC014602	A
Pseudoxylomyces elegans	IS	KT 2887	MAFF 243852	HHUF 30139	AB797308	AB807598	AB808576	LC014593	C

¹Abbreviation of family names: Bam (Bambusicolaceae), Dic (Dictyosporaceae), Did (Didymosphaeriaceae), IS (insertae sedis), Len (Lentitheciaceae), Mas (Massarinaceae), Mor (Morosphaeriaceae), Par (Paramorosphaeriaceae), Per (Periconiaceae), Sul (Sulcatisporaceae), and Tre (Trematosphaeriaceae).

²Specimen with HT (holotype), PT (paratype), and ET (epitype).

³Sequences obtained in this study are shown in bold. Sequences with¹ from Tanaka et al. (2009),² from Schoch et al. (2009), and ³ from Hirayama et al. (2010).

⁴Origin of isolates: A, single ascospore; C, single conidium.

Molecular phylogenetic analysis

A total of 106 isolates were used for DNA extraction (Table 1). DNA from mycelia was extracted using the ISOPLANT Kit (Nippon Gene, Tokyo, Japan) following the manufacturer’s instructions. Partial SSU and LSU nrDNA, and tef1 were sequenced to elucidate phylogenetic relationships of the isolates for considering familial and generic classifications. The complete internally transcribed spacer (ITS) regions of nrDNA were also obtained for use as DNA barcode markers (Schoch et al. 2012), although the sequences were not used for phylogenetic reconstruction (Table 1). Four primer sets, NS1–NS4 (White et al. 1990), LR0R–LR7 (Rehner & Samuels 1994), EF1-983F–EF1-2218R (Rehner & Buckley 2005), and ITS1–ITS4 (White et al. 1990) were used for the amplification of SSU, LSU, tef1, and ITS, respectively. DNA fragments were amplified and sequenced following the methods described by Tanaka et al. (2009). Newly obtained sequences have been deposited in GenBank (Table 1). These sequences together with those retrieved from GenBank (Table 2) were aligned by MUSCLE included in the program Molecular Evolutionary Genetic Analysis (MEGA) v. 6 (Tamura et al. 2013) and manually adjusted to optimise the alignment. Hysterobrevium mori and Hysterium pulicare, both belonging to the Hysteriales, were designated as outgroup taxa. The alignments used were deposited in TreeBASE (http://www.treebase.org). Phylogenetic analyses were conducted based on maximum likelihood (ML) method. The optimum substitution models for each dataset were estimated by Kakusan4 (Tanabe 2011), based on the Akaike information criterion (AIC; Akaike 1974) for ML analyses. The ML analyses were performed with TreeFinder Mar 2011 (Jobb 2011) based on the models selected by AIC (separate model among genes and proportional model among codons, HKY85+G for SSU, J2+G for LSU, GTR+G for the first codon of tef1, TVM+G for the second codon of tef1, and GTR+G for the third codon of tef1). Bootstrap proportions (BP) were obtained by 1000 bootstrap replications.

Table 2.

Cultures and GenBank accession numbers of Massarineae used for phylogenetic analysis.

Taxon	Family1	Culture no.2	GenBank accession no.3
SSU	LSU	tef1
Alloconiothyrium aptrootii	Did	CBS 980.95HT	NS	JX496234	NS
Aquaticheirospora lignicola	Dic	RK-2006aHT	AY736377	AY736378	NS
Aquilomyces patris	Mor	CBS 135661HT	KP184077	KP184041	NS
Asteromassaria pulchra	IS	CBS 124082	GU296137	GU301800	GU349066
Bambusicola bambusae	Bam	MFLUCC 11-0614HT	JX442039	JX442035	NS
B. irregulispora	Bam	MFLUCC 11-0437HT	JX442040	JX442036	NS
B. loculata	Bam	MFLUCC 13-0856HT	KP761735	KP761729	KP761724
B. massarinia	Bam	MFLUCC 11-0389HT	JX442041	JX442037	NS
B. splendida	Bam	MFLUCC 11-0439HT	JX442042	JX442038	NS
Bambusistroma didymosporum	Per	MFLUCC 13-0862HT	NS	KP761730	KP761727
Bimuria novae-zelandiae	Did	CBS 107.79HT	AY016338	AY016356	DQ471087
Byssothecium circinans	Mas	CBS 675.92	GU205235	AY016357	GU349061
Camarographium koreanum	Mac	CBS 117159HT	NS	JQ044451	NS
“Corynespora” leucadendri	Mas	CBS 135133HT	NS	KF251654	NS
“C.” olivacea	Mas	CBS 114450	NS	GU301809	GU349014
Cucurbidothis pityophila	Did	CBS 149.32	U42480	DQ384102	NS
Darksidea alpha	Len	CBS 135650HT	KP184049	KP184019	KP184166
D. beta	Len	CBS 135637HT	KP184074	KP184023	KP184189
D. gamma	Len	CBS 135634HT	KP184073	KP184028	KP184188
D. delta	Len	CBS 135638HT	KP184069	KP184024	KP184184
D. epsilon	Len	CBS 135658HT	KP184070	KP184029	KP184186
D. zeta	Len	CBS 135640HT	KP184071	KP184013	KP184191
Dendryphiella vinosa	Dic	–	EU848589	EU848590	NS
Deniquelata barringtoniae	Did	MFLUCC 110422HT	JX254656	JX254655	NS
Dictyosporium alatum	Dic	ATCC 34953HT	DQ018080	DQ018101	NS
D. elegans	Dic	NBRC 32502	DQ018079	DQ018100	NS
D. inflatum	Dic	NTOU 3855	JQ267361	JQ267363	NS
D. meiosporum	Dic	MFLUCC 10-0131HT	KP710946	KP710945	NS
D. stellatum	Dic	CCFC 241241HT	NS	JF951177	NS
D. strelitziae	Dic	CBS 123359HT	NS	FJ839653	NS
D. thailandicum	Dic	MFLUCC 13-0773HT	NS	KP716707	NS
D. toruloides	Dic	CBS 209.65	DQ018081	DQ018104	NS
Didymocrea sadasivanii	Did	CBS 438.65IT	DQ384066	DQ384103	NS
Didymosphaeria rubi-ulmifolii	Did	MFLUCC 14-0023HT	KJ436588	KJ436586	NS
“D.” spartii	Mas	CBS 183.58	GU205250	GU205225	NS
Digitodesmium bambusicola	Dic	CBS 110279HT	NS	DQ018103	NS
“Diplococcium” asperum	Dic	CBS 139.95	EF204511	EF204493	NS
Falciformispora lignatilis	Tre	BCC 21117	GU371834	GU371826	GU371819
F. senegalensis	Tre	CBS 196.79HT	KF015636	KF015631	KF015687
F. tompkinsii	Tre	CBS 200.79HT	KF015639	KF015625	KF015685
Flavomyces fulophazii	Per	CBS 135761HT	KP184082	KP184040	NS
Halomassarina thalassiae	Tre	BCC 17054	GQ925842	GQ925849	NS
H. thalassiae	Tre	JK 5262D	NS	GU301816	GU349011
Helicascus aegyptiacus	Mor	FWCC 99HT	KC894852	KC894853	NS
H. aquaticus	Mor	MFLUCC 10-0918HT	KC886638	KC886640	NS
H. elaterascus	Mor	A22-5A = HKUCC 7769	AF053727	AY787934	NS
H. nypae	Mor	BCC 36751	GU479754	GU479788	GU479854
H. nypae	Mor	BCC 36752	GU479755	GU479789	GU479855
H. thalassioideus	Mor	MFLUCC10-0911	KC886637	KC886636	NS
H. unilocularis	Mor	MJF14020HT	NS	KP637166	NS
Hysterium pulicare	OG	CBS 12337	FJ161161	FJ161201	FJ161109
Hysterobrevium mori	OG	CBS 12356	FJ161155	FJ161196	FJ161104
Inflatispora pseudostromatica	IS	CBS 123110HT	JN231132	JN231131	NS
“Kalmusia (Montagnula)” anthostomoides	Did	CBS 615.86	GU205246	GU205223	NS
K. ebuli	Did	CBS 123120NT	JN851818	JN644073	NS
K. italica	Did	MFLUCC 13–0066HT	KP325442	KP325441	NS
K. spartii	Did	MFLUCC 14–0560HT	KP753953	KP744487	NS
K. variispora	Did	CBS 121517HT	NS	JX496143	NS
Karstenula rhodostoma	Did	CBS 690.94	GU296154	GU301821	GU349067
Keissleriella cladophila	Len	CBS 104.55	GU296155	GU301822	GU349043
K. genistae	Len	CBS 113798	GU205242	GU205222	NS
K. linearis	Len	IFRD 2008	FJ795478	FJ795435	NS
K. poagena	Len	CBS 136767HT	NS	KJ869170	NS
K. sparticola	Len	MFLUCC 14–0196HT	NS	KP639571	NS
K. trichophoricola	Len	CBS 136770HT	NS	KJ869171	NS
Latorua caligans	Lat	CBS 576.65HT	NS	KR873266	NS
L. grootfonteinensis	Lat	CBS 369.72HT	NS	KR873267	NS
Lentithecium aquaticum	Len	CBS 123099HT	GU296156	GU301823	GU349068
L. fluviatile	Len	CBS 122367	GU296158	GU301825	GU349074
Leptosphaeria doliolum	Lep	CBS 505.75	GU296159	GU301827	GU349069
Letendraea eurotioides	Did	CBS 212.31	NS	AY787935	NS
L. helminthicola	Did	CBS 884.85	AY016345	AY016362	NS
L. padouk	Did	CBS 485.70	GU296162	AY849951	NS
Lophiostoma arundinis	Lop	JCM 13550	AB618679	AB618998	LC001737
L. macrostomum	Lop	JCM 13544	AB618691	AB619010	LC001751
Macrodiplodiopsis desmazieri	Mac	CBS 140062ET	NS	KR873272	NS
M. desmazieri	Mac	CBS 221.37	DQ678013	DQ678065	DQ677908
Massarina eburnea	Mas	CBS 473.64	GU296170	GU301840	GU349040
Montagnula aloes	Did	CBS 132531HT	NS	JX069847	NS
M. graminicola	Did	MFLUCC 13–0352HT	KM658316	KM658315	NS
M. opulenta	Did	CBS 168.34	AF164370	DQ678086	NS
Morosphaeria ramunculicola	Mor	BCC 18404	GQ925838	GQ925853	NS
M. velatispora	Mor	BCC 17059	GQ925841	GQ925852	NS
Multiseptospora thailandica	Par	MFLUCC 11–0183HT	KP753955	KP744490	NS
Munkovalsaria appendiculata	Did	CBS 109027HT	NS	AY772016	NS
Murilentithecium clematidis	Len	MFLUCC 14-0561HT	KM408761	KM408759	KM454445
Neobambusicola strelitziae	Sul	CBS 138869HT	NS	KP004495	NS
Noosia banksiae	Per	CBS 129526HT	NS	JF951167	NS
Palmiascoma gregariascomum	Bam	MFLUCC 11–0175HT	KP753958	KP744495	NS
Paracamarosporium leucandendri	Did	CBS 123027HT	NS	EU552106	NS
P. psoraleae	Did	CBS 136628HT	NS	KF777199	NS
Paraconiothyrium estuarinum	Did	CBS 109850HT	NS	JX496129	NS
“P.” flavescens	Dic	CBS 178.93	GU238216	GU238075	NS
“P.” fuckelii	Did	CBS 797.95	GU238204	GU237960	NS
P. maculicutis	Did	CBS 101461HT	EU754101	EU754200	NS
P. thysanolaenae	Did	MFLUCC 10–0550HT	KP753959	KP744496	NS
“P.” tiliae	Did	CBS 265.94LT	EU754040	EU754139	NS
Paraphaeosphaeria michotii	Did	CBS 652.86	GQ387520	GQ387581	GU456266
“P.” spartii	Did	MFLUCC 13–0398HT	KP711367	KP711362	NS
Periconia macrospinosa	Per	CBS 135663	KP184080	KP184038	NS
Phaeodothis winteri	Did	CBS 182.58	GU296183	GU301857	DQ677917
Phragmocamarosporium hederae	Len	MFLUCC 13-0552HT	KP842918	KP842915	NS
P. platani	Len	MFLUCC 14-1191HT	KP842919	KP842916	NS
“Pithomyces” valparadisiacus	Mor	CBS 113339	NS	EU552152	NS
Pleospora herbarum	Ple	CBS 191.86	DQ247812	DQ247804	DQ471090
Pleurophoma pleurospora	Len	CBS 130329 LT	NS	JF740327	NS
Poaceascoma helicoides	Len	MFLUCC 11-0136HT	KP998463	KP998462	KP998461
Polyschema congolensis	Lat	CBS 542.73HT	NS	EF204502	NS
P. larviformis	Lat	CBS 463.88	NS	EF204503	NS
P. terricola	Lat	CBS 301.65HT	EF204519	EF204504	NS
Pseudocamarosporium brabeji	Did	CBS 119219HT	NS	EU552104	NS
P. cotinae	Did	MFLUCC 14–0624HT	KP753964	KP744505	NS
P. propinquum	Did	MFLUCC 13-0544ET	KJ819949	KJ813280	NS
P. tilicola	Did	MFLUCC 13-0550HT	KJ819950	KJ813281	NS
Pseudochaetosphaeronema larense	Mac	CBS 640.73HT	KF015652	KF015611	KF015684
Pseudodictyosporium elegans	Dic	CBS 688.93HT	DQ018084	DQ018106	NS
P. wauense	Dic	NBRC 30078	DQ018083	DQ018105	NS
Setoseptoria arundinacea	Len	CBS 619.86	GU296157	GU301824	NS
S. arundinacea	Len	CBS 123131	GU456298	GU456320	GU456281
S. phragmitis	Len	CBS 114802HT	NS	KF251752	NS
S. phragmitis	Len	CBS 114966PT	NS	KF251753	NS
Sporidesmiella fusiformis	Did	HKUCC 10831	NS	DQ408577	NS
“Sporidesmium” tengii	Per	HKUCC 10837	NS	DQ408559	NS
Stagonospora duoseptata	Mas	CBS 135093HT	NS	KF251758	NS
“S.” macropycnidia	Len	CBS 114202	GU296198	GU301873	GU349026
S. paludosa	Mas	CBS 135088NT	NS	KF251760	NS
S. (Neottiosporina) paspali	Mas	CBS 331.37	EU754073	EU754172	GU349079
S. perfecta	Mas	CBS 135099HT	NS	KF251761	NS
S. pseudocaricis	Mas	CBS 135132HT	NS	KF251763	NS
S. pseudopaludosa	Mas	CBS 136424HT	NS	KF777239	NS
S. pseudovitensis	Mas	S602PT	NS	KF251765	NS
S. trichophoricola	Mas	CBS 136764HT	NS	KJ869168	NS
S. uniseptata	Mas	CBS 135090HT	NS	KF251767	NS
Suttonomyces clematidis	Mas	MFLUCC 14-0240HT	KP842920	KP842917	NS
Trematosphaeria grisea	Tre	CBS 332.50HT	KF015641	KF015614	KF015698
T. grisea	Tre	CBS 135984	KF015632	KF015618	KF015694
T. pertusa	Tre	CBS 122368ET	FJ201991	FJ201990	KF015701

¹Abbreviation of family names: Acr (Acrocalymmataceae), Bam (Bambusicolaceae), Dic (Dictyosporaceae), Did (Didymosphaeriaceae), IS (insertae sedis), Lat (Latoruaceae), Len (Lentitheciaceae), Lep (Leptosphaeriaceae), Lop (Lophiostomataceae), Mac (Macrodiplodiopsidaceae), Mas (Massarinaceae), Mor (Morosphaeriaceae), OG (outgroup), Par (Parabambusicolaceae), Per (Periconiaceae), Ple (Pleosporaceae), Sul (Sulcatisporaceae), and Tre (Trematosphaeriaceae).

²Isolate ex ET (epitype), HT (holotype), IT (isotype), LT (lectotype), NT (neotype), and PT (paratype).

³NS: No sequence available in GenBank.

Results

Molecular phylogenetic analysis

Approximately 940–1 750 bp of SSU, 870–1 330 bp of LSU nrDNA, 830–940 bp of tef1, and 500–900 bp of ITS sequences were determined for 106 isolates of fungi within the Massarineae. Analyses of different gene datasets were performed individually, but no topological conflict was observed at familial level with the exception of Lentitheciaceae, which was polyphyletic in the tef1 tree (data not shown). A combined dataset of SSU, LSU, and tef1 sequences was generated after excluding insertions of several species which corresponded to positions 493–1 005 of Monodictys capensis (GenBank AB797261) and positions 1 286–1 651 of Magnicamarosporium iriomotense (GenBank AB797219) in the SSU, and positions 836–892 of Montagnula spartii (GenBank GU205225) and positions 871–924 of Hysterium pulicare (GenBank FJ161201) in the LSU sequences. The combined dataset consisted of 243 taxa and 3 386 characters, of which 28 % were missing and gap characters. The alignment had 82 % representation for SSU, 100 % for LSU and 60 % for tef1. BP support of each familial clade in the LSU tree was generally improved by adding the SSU and tef1 dataset with missing data, e.g., from 82 % to 100 % in the Massarinaceae, and from 90 % to 100 % in the Trematosphaeriaceae.

The ML tree of Massarineae based on the SSU and LSU nrDNA and tef1 regions with the highest log likelihood (−36965.05250) is shown in Fig. 1. A total of 237 taxa of the Massarineae formed a clade (with 98 % BP support) and were scattered in 12 familial clades and five unknown clades. Seven families previously recognised, namely the Dictyosporiaceae (nom. prov., 100 % BP), Didymosphaeriaceae (99 % BP), Latoruaceae (99 % BP), Macrodiplodiopsidaceae (96 % BP), Massarinaceae (100 % BP), Morosphaeriaceae (98 % BP), and Trematosphaeriaceae (100 % BP) were highly supported as independent monophyletic groups. The Bambusicolaceae and Lentitheciaceae received moderate BP support, 87 % and 71 %, respectively. We erected two new families, the Parabambusicolaceae (81 % BP) and Sulcatisporaceae (97 % BP), to accommodate several genera, which cannot be placed in any of the existing families in the Massarineae. Although species in the genus Periconia have been treated as members of the Massarinaceae (Zhang et al., 2012, Hyde et al., 2013), we have placed them in a distinct family, the Periconiaceae (100 % BP), which was established by Nannizzi (1934).

Fig. 1.

Maximum-likelihood tree of Massarineae based on the SSU and LSU nrDNA and tef1 regions. ML bootstrap proportion (BP) greater than 50 % is presented at the nodes. An original isolate designation (or culture collection number) is noted after the species name. Sequences derived from holotype, isotype, neotype, paratype and epitype materials are indicated as HT, IT, NT, PT and ET, respectively. Species used for morphological observation in this study are formatted in bold. New names are marked by five-pointed stars. Families, where known, are indicated with coloured blocks. The tree was rooted to Hysterobrevium mori and Hysterium pulicare in the Hysteriales. The scale bar represents the number of nucleotide substitutions per site.

Taxonomy

As a result of morphological comparisons and phylogenetic analyses of 106 strains, along with sequences from 131 taxa obtained from GenBank, at least 12 families including two new families (the Parabambusicolaceae and Sulcatisporaceae) are recognised. Ten new genera, 22 new species, and seven new combinations are proposed. Taxa are arranged in alphabetical order by family, genus, and species. For the known species a brief description and/or taxonomic notes are provided.

Dictyosporiaceae nom. prov. (see Liu et al. 2015)

Type genus: Dictyosporium Corda.

Dictyosporium Corda, Weitenweber's Beitr. Nat.: 87. 1836.

Type species: Dictyosporium elegans Corda.

Notes: Dictyosporium species have been reported worldwide from dead wood and decaying leaves in terrestrial and freshwater environments. The genus is characterised by pigmented, cheiroid conidia formed on sporodochial colonies. It has been considered that the genus is closely related to the Massarinaceae in the Pleosporales based on phylogenetic analysis using SSU and LSU nrDNA sequences (Tsui et al. 2006). After taxonomic revision of the genus (Goh et al., 1999, Cai et al., 2003), 33 species were accepted as Dictyosporium (Crous et al. 2009a). In addition, 16 species have been further recorded in this genus (Manoharachary et al., 2007, Zhang et al., 2009a, Wongsawas et al., 2009, Hu et al., 2010, McKenzie, 2010, Crous et al., 2011a, Whitton et al., 2012, Kirschner et al., 2013, Liu et al., 2015, Prasher and Verma, 2015).

Dictyosporium bulbosum Tzean & J.L. Chen, Mycol. Res. 92: 500. 1989. Fig. 2A–C.

Fig. 2.

Dictyosporium spp. A–C. D. bulbosum (A from culture yone 221; B, C from yone 221); D–F. D. aff. bulbosum (culture KH 375); G–I. D. digitatum (G from culture KT 2660; H from culture yone 280; I from culture KH 401); J–L. D. aff. heptasporum (KH 332); M–O. D. hughesii (M, O from KT 1847; N from culture KT 1847); P–V. D. pseudomusae (P from yone 234; Q, T from culture yone 234; R, U from culture KH 412; S, V from KH 412); W–Y. D. tetrasporum (culture KT 2865). A, D, G, J, M, P–R, W. Conidial masses on culture media or natural substrates (A, D, G, Q, R, W from culture; J, M, P from nature); B, C, E, F, H, I, K, L, N, O, S–V, X, Y. Conidia (arrowheads indicate conidial appendages). Scale bars: A, D, G, J, M, P–R, W = 200 μm; B, C, E, F, H, I, K, L, N, O, S–V, X, Y = 10 μm.

Specimen examined: Japan, Okinawa, Isl. Iriomote, near Maryudu-falls, on dead twigs of woody plant, 27 Sep. 2007, H. Yonezawa & K. Tanaka, yone 221 = HHUF 29990, culture MAFF 243835.

Note: This specimen was identified as D. bulbosum and morphological features were described and illustrated by Hirayama et al. (2012).

Dictyosporium aff. bulbosum Fig. 2D–F.

Specimen examined: Japan, Okinawa, Isl. Iriomote, Uehara, near Tropical Biosphere Research Center (Ryukyu Univ.), small stream, on submerged twigs of woody plant, 13 Jul. 2011, K. Hirayama & K. Tanaka, KH 375 = HHUF 30127, culture JCM 19403 = MAFF 243829.

Notes: The above isolate produced an asexual morph in culture, which is similar to that on the natural specimen. Conidia in culture were 36–50 × 26–33 μm (av. 45.7 × 29.1 μm, n = 33), l/w 1.3–1.8 (av. 1.6, n = 33), consisting of 29–47 cells arranged in 5–6 rows, with apical appendages. These features almost agree with the description in the protologue of D. bulbosum (Tzean & Chen 1989), but the ITS sequences of our material (KH 375; GenBank LC014542) differed from those of D. bulbosum (GenBank LC014544 and DQ018086) in ca. 8 % (40/515) positions.

Dictyosporium digitatum J.L. Chen et al., Mycol. Res. 95: 1145. 1991. Fig. 2G–I.

Specimens examined: Japan, Okinawa, Isl. Ishigaki, trail of Mt. Omoto, on dead stems of herbaceous plant, 16 Jul. 2011, K. Hirayama & K. Tanaka, KH 401 = HHUF 30128, culture JCM 19404 = MAFF 243830; Okinawa, Isl. Iriomote, Komi, on dead wood of Castanopsis sieboldii, 15 Sep. 2009, Y. Kurihara, KT 2660 = HHUF 30131, culture JCM 19405 = MAFF 243833; Okinawa, Isl. Iriomote, Kanpire-falls, on dead twigs of woody plant, 21 Nov. 2008, K. Tanaka & K. Hirayama, yone 280 = HHUF 30093, culture MAFF 243837.

Notes: The morphological characters of our specimens are consistent with those of D. digitatum (Chen et al. 1991), as reported by Hirayama et al. (2012). The ITS sequences of this species from our three isolates were highly similar (99.1–99.8 %) but their similarities with a deposited sequence of D. digitatum in GenBank (GenBank DQ018089) were rather low (90.1–90.4 %).

Dictyosporium aff. heptasporum Fig. 2J–L.

Specimen examined: Japan, Okinawa, Isl. Ishigaki, Mt. Banna, near small stream, on dead twigs of woody plant, 14 Jul. 2011, K. Hirayama & K. Tanaka, KH 332 = HHUF 30126, culture JCM 19406 = MAFF 243828.

Notes: The morphological features of the above specimen were as follows; conidia 70–90 × 21–31 μm (av. 79.7 × 23.3 μm, n = 43), cylindrical, l/w 3.1–3.9 (av. 3.4, n = 43), with (5–)7 rows, 16–18-septate, without appendages. This agrees with the details in the description of D. heptasporum (Goh et al. 1999), but the conidia in our material are somewhat larger (vs. 50–80 × 20–30 μm; Goh et al. 1999). A BLAST search using ITS sequences from our culture showed D. heptasporum (GenBank DQ018090) as the closest species, but the similarity was relatively low (493/518 = 95.2 %).

Dictyosporium hughesii McKenzie, Mycotaxon 111: 156. 2010. Fig. 2M–O.

Specimen examined: Japan, Kagoshima, Isl. Yakushima, Shirataniunsuikyo, on dead twigs of Stewartia monadelpha, 18 Oct. 2005, K. Tanaka & T. Hosoya, KT 1847 = HHUF 30130 = TNS-F-12407, culture JCM 19407 = MAFF 243832.

Note: This collection was identified as D. hughesii (McKenzie 2010) based on the conidial morphology; conidia 43–51 × 18–28 μm (av. 46.8 × 24.8 μm, n = 30), l/w 1.6–2.3(–2.6) (av. 1.9, n = 30), consisting of 50–71 cells arranged in 6–7 rows, with or without apical appendages.

Dictyosporium pseudomusae Kaz. Tanaka, G. Sato & K. Hiray., sp. nov. MycoBank MB811297. Fig. 2P–V.

Etymology: After its morphological similarity to Dictyosporium musae.

Sporodochia on natural substrate scattered, punctiform, dark brown to black. Mycelium immersed, 170–490 μm diam. Conidiophores micronematous, not differentiated from vegetative hyphae. Conidiogenous cells holoblastic, cylindrical, 5.5–8 μm wide. Conidia solitary, brown, ellipsoid to cylindrical, cheiroid, not complanate, (58–)61–78(–81) × 19–29(–33) μm (av. 69.9 × 22.9 μm, n = 60), l/w (2.2–)2.5–3.8(–4.0) (av. 3.1, n = 60), consisting of 78–100 cells arranged in (6–)7 rows and basal connecting cell (6–8 × 5.5–8 μm); each row cylindrical, with 13–15 cells. Appendages globose to subglobose, hyaline, 6–11.5 μm diam, bearing from apical cells or side of outer rows. Sexual morph unknown.

Colonies on PDA (after 4 wk) attaining a diam of 3.9–4.4 cm, white to rosy buff; reverse buff to cinnamon; rosy vinaceous pigment produced. In culture asexual morph formed.

Specimens examined: Japan, Okinawa, Isl. Ishigaki, Mt. Banna, near small stream, on dead twigs of Bamboo, 14 Jul. 2011, K. Hirayama & K. Tanaka, KH 412 = HHUF 30129, culture JCM 19408 = MAFF 243831; Okinawa, Isl. Iriomote, Inamori-path, on dead twigs of woody plant, 25 Sep. 2007, H. Yonezawa & K. Tanaka (holotype yone 234 = HHUF 30133, culture ex-type CBS 139686 = JCM 19409 = MAFF 243836).

Notes: Among the described species, D. pseudomusae is most similar to D. musae (Photita et al. 2002) in having large-sized conidia comprised of seven cell rows and with appendages arising from side cells of the outer rows. Conidia of the latter species, however, are smaller (45–65 × 20–27 μm, av. 55.9 × 23.5 μm) and comprised of fewer cells (49–77 cells; Photita et al. 2002). The closest species to D. pseudomusae appear to be D. digitatum [GenBank DQ018089; identities = 504/516 (97.7 %), gaps = 2/516] and D. giganticum [GenBank DQ018095; identities = 497/516 (96.3 %), gaps = 2/516] from a BLAST search using ITS sequences.

Dictyosporium tetrasporum L. Cai & K.D. Hyde, Mycoscience 48: 290. 2007. Fig. 2W–Y.

Specimen examined: Japan, Okinawa, Isl. Ishigaki, Mt. Banna, near small stream, on dead twigs of woody plant, 16 Jul. 2011, K. Tanaka & K. Hirayama, KT 2865 = HHUF 30132, culture JCM 19410 = MAFF 243834.

Notes: The collection matches the original description of D. tetrasporum (Cai & Hyde 2007a). The characters of our specimen are as follows; conidia 22–37 × 15–21 μm (av. 27.5 × 17.5 μm, n = 30), l/w 1.2–2.2 (av. 1.6, n = 30), consisting of 11–28 cells arranged in (3–)4 rows, without apical appendages. In culture, conidia were produced that were identical to those on the natural specimen (24–38 × 16–24 μm; av. 30.3 × 20.1 μm, n = 32).

Gregarithecium Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811298.

Etymology: Referring to the gregarious ascomata.

Ascomata grouped, immersed to erumpent, depressed globose to hemispherical with flattened base in section. Ostiolar neck short terete, central, with periphyses, covered by black clypeus. Ascomatal wall composed of thin-walled cells, surrounded by vertically-orientated stromatic tissue. Pseudoparaphyses septate, branched and anastomosed. Asci fissitunicate, cylindrical, short-stalked, with 8 biseriate ascospores. Ascospores broadly fusiform, with a median septum, hyaline, smooth, surrounded by an entire sheath. Asexual morph unknown.

Type species: Gregarithecium curvisporum Kaz. Tanaka & K. Hiray.

Notes: In its depressed globose ascomata with clypeate ostiole, Gregarithecium is reminiscent of species in Massarina s. lat. (Aptroot, 1998, Tanaka and Harada, 2003b), but Gregarithecium can be separated from the latter by the presence of stromatic tissue surrounding the ascomata. Massarina s. lat. is regarded as being polyphyletic and thus has recently been segregated into several new genera. These are scattered in the Lentitheciaceae (Tingoldiago; Hirayama et al. 2010), Morosphaeriaceae (Morosphaeria; Suetrong et al. 2009), Trematosphaeriaceae (Halomassarina; Suetrong et al. 2009), Tetraplosphaeriaceae (Triplosphaeria; Tanaka et al. 2009), and Lindgomycetaceae (Lindgomyces; Hirayama et al. 2010), but are not so far reported in the Dictyosporiaceae.

Gregarithecium curvisporum Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811299. Fig. 3.

Fig. 3.

Gregarithecium curvisporum. A, B. Ascomata on the natural host surface; C. Ascomata in culture (on rice straw); D, E. Ascomata in longitudinal section; F. Ascomatal wall; G. Pseudoparaphyses; H. Ascus; I–K. Ascospores (arrowheads indicate mucilaginous sheath); L. Spermatia. A, B, D–I from KT 922; C, J–L from culture KT 922. Scale bars: A = 2 mm; B, C = 500 μm; D, E = 100 μm; F–L = 10 μm.

Etymology: From the Latin curvi-, in reference to the curved ascospores.

Ascomata grouped in numbers of 3–6, immersed to erumpent, depressed globose to hemispherical with flattened base in section, 140–180 μm high, 290–430 μm diam. Ostiolar neck short terete, central, with an ostiole (25–50 μm diam) and short periphyses, covered by a black clypeus composed of small cells (2.5–7.5 × 1–3 μm). Ascomatal wall in longitudinal section 8–12 μm thick at the sides, composed of thin-walled, flattened, pale brown cells, surrounded by vertically-oriented stromatic tissue composed of rectangular to polygonal cells (5–25 × 5–16 μm) or by compact brown hyphae. Pseudoparaphyses septate, branched and anastomosed, 2–3.5(–5) μm wide. Asci fissitunicate, cylindrical, 73.5–102 × 11–15.5 μm (av. 87.4 × 13.6 μm, n = 50), rounded at the apex, with a shallow ocular chamber (0.5–1 μm high), short-stalked (5–15 μm long; av. 9.6 μm, n = 32), with 8 biseriate ascospores. Ascospores broadly fusiform, mostly curved, 19–31 × 4.5–6.5 μm (av. 24.2 × 5.8 μm, n = 50), l/w (3.3–)3.6–4.9(–5.2) (av. 4.2, n = 50), with a median (0.50) septum and constricted, hyaline, smooth, with an entire sheath; sheath sharply delimited and 1–2 μm wide at first, diffuse and 4–8 μm wide at a later. Senescent ascospores 3-septate, thick-walled, dark brown.

Colonies on PDA (after 4 wk) attaining a diam of 6.2–6.8 cm, buff to smoke grey; reverse pale luteous to sienna; no pigment produced. In culture ascomatal and spermatial morphs observed. Asci 82.5–101.5 × 14.5–17.5 μm. Ascospores 21–31 × 6–8 μm (av. 24.5 × 7.0 μm, n = 31), l/w 3.1–4.2 (av. 3.5, n = 31). Spermatia 2.2–3.5 × 1.4–2.1 μm (av. 3.0 × 1.8 μm, n = 20), subglobose, hyaline.

Specimen examined: Japan, Aomori, Hirosaki, Kozawa, on dead culms of Sasa sp., 25 Oct. 2002, T. Handa (holotype KT 922 = HHUF 30134, culture ex-type CBS 139688 = JCM 19411 = MAFF 243838).

Pseudocoleophoma Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811300.

Etymology: Referring to the similarity of the asexual morph with that of Coleophoma.

Ascomata scattered to grouped, immersed to erumpent, globose to subglobose in section. Ostiolar neck central, composed of subglobose dark brown cells. Ascomatal wall composed of polygonal to rectangular cells. Pseudoparaphyses numerous. Asci fissitunicate, cylindrical to clavate, short-stalked, with 8 ascospores. Ascospores fusiform, 1-septate, smooth, with a conspicuous sheath. Conidiomata coleophoma-like, pycnidial, subglobose. Conidiophores absent. Conidiogenous cells phialidic, doliiform to lageniform. Conidia cylindrical, hyaline, smooth.

Type species: Pseudocoleophoma calamagrostidis Kaz. Tanaka & K. Hiray.

Notes: The pycnidial asexual morph of Pseudocoleophoma bears a slight resemblance to that of Coleophoma. The type species of Coleophoma (C. crateriformis), however, has pycnidia possessing paraphyses that are not found in Pseudocoleophoma, and is a member of the Dothideales, rather than the Pleosporales (De Gruyter et al. 2009).

Pseudocoleophoma calamagrostidis Kaz. Tanaka & K. Hiray., sp. nov. MB811301. Fig. 4.

Fig. 4.

Pseudocoleophoma calamagrostidis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall at side; E. Ascomatal wall surface around ostiole; F. Pseudoparaphyses; G, H. Asci; I–K. Ascospores (arrowheads indicate gelatinous sheath; I. in Black-Blue ink); L. Germinating ascospore; M, N. Conidiomata in culture (on rice straw); O. Conidioma in longitudinal section; P. Conidiomatal wall at side; Q. Conidiomatal wall around ostiole; R. Conidiogenous cell; S. Conidia. A–L from KT 3284; M–S from culture KT 3284. Scale bars: A, B, M, N = 500 μm; C, E, O = 50 μm; D, F–L, P–S = 10 μm.

Etymology: Referring to the host plant.

Ascomata scattered, immersed, erumpent at the top, globose to depressed globose, 140–200 μm high, 160–220 μm diam, ostiolate. Ostiolar neck central. Ascomatal wall in longitudinal section 5–10 μm thick at sides, composed of 2–3 layers of thin-walled, polygonal, flattened, 5–12 × 2–5 μm, brown cells; of polygonal to subglobose, 2–5 × 1.5–2.5 μm cells around ostiole. Pseudoparaphyses septate, branched and anastomosed, 2.5–5 μm wide. Asci fissitunicate, cylindrical, 62.5–80 × 7.5–10 μm (av. 69.0 × 8.4 μm, n = 50), rounded at the apex, with a shallow ocular chamber, short-stalked (5–12 μm long), with 8 biseriate ascospores. Ascospores narrowly fusiform, (14.5–)16–19(–21) × 3–4.5 μm (av. 17.4 × 3.8 μm, n = 30), l/w 4.0–5.3 (av. 4.5, n = 30), with a nearly median (0.43–0.54, av. 0.50, n = 26) septum, constricted at the septum, hyaline, smooth, with an entire sheath; sheath sharply delimited, 1–2 μm wide at sides, 4–6 μm long at both ends, staining with Black-Blue ink when in fresh condition.

Colonies on PDA (after 4 wk) attaining a diam of 2.3–3.2 cm, buff to honey; reverse similar; no pigment produced. In culture coelomycetous asexual morph formed. Conidiomata pycnidial, 220–300 μm high, (150–)250–500 μm diam, immersed to erumpent, depressed globose. Ostiolar neck well-developed, 75–100 μm long, 85–100 μm wide, cylindrical, central. Conidiomatal wall in longitudinal section uniformly 7.5–15 μm thick, composed of 3–4 layers of slightly thick-walled, polygonal to subglobose, 6–15 × 2–6.5 μm, pale brown cells; of polygonal to rectangular, 2–6 × 1.5–2.5 μm cells around ostiole. Conidiophores absent. Conidiogenous cells phialidic, 5–9 × 2–4 μm, doliiform to subglobose. Conidia cylindrical, aseptate, hyaline, smooth, 6–10 × 2–2.5 μm (av. 8.6 × 2.2 μm, n = 50), l/w 2.9–4.7 (av. 3.9, n = 50).

Specimen examined: Japan, Aomori, Hirosaki, Mt Iwaki, on dead leaves of Calamagrostis matsumurae, 27 Jul. 2013, K. Tanaka (holotype KT 3284 = HHUF 30450, culture ex-type CBS 139700).

Note: This species is phylogenetically close to P. polygonicola, but the ITS sequence similarity between these two species is relatively low (490/521 = 94.0 %).

Pseudocoleophoma polygonicola Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811302. Fig. 5.

Fig. 5.

Pseudocoleophoma polygonicola. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–J. Ascospores (arrowheads indicate gelatinous sheath; J. in Black-Blue ink); K. Conidiomata in culture; L. Conidioma in longitudinal section; M. Conidiomatal wall; N. Conidiogenous cells; O. Conidia. A–J from KT 731; K–O from culture KT 731. Scale bars: A = 2 mm; B, K = 500 μm; C, L = 50 μm; D–J, M–O = 10 μm.

Etymology: Referring to the host plant.

Ascomata scattered to 2–4 grouped, immersed to erumpent, 230–310 μm high, 280–350 μm diam. Ostiolar neck central, 50–75 μm long, 70–100 μm wide. Ascomatal wall in longitudinal section 7–13 μm thick at sides. Pseudoparaphyses 2–2.5 μm wide. Asci cylindrical to clavate, (67–)74–90(–100) × 9–12.5 μm (av. 81.3 × 10.7 μm, n = 48). Ascospores fusiform, (17.5–)19–23(–25) × 4–6 μm (av. 20.6 × 4.8 μm, n = 50), l/w 3.6–4.8 (av. 4.2, n = 45), with a septum supramedian (0.46–0.50; av. 0.48, n = 40), surrounded by a sheath; sheath 1–2 μm wide at sides, 2–5 μm long at both ends, staining with Black-Blue ink when in fresh condition.

Colonies on PDA (after 4 wk) attaining a diam of 3.4–4.8 cm, white; reverse white to straw; no pigment produced. In culture coelomycetous asexual morph formed. Conidiomata pycnidial, 170–250 μm diam. Conidiomatal wall in longitudinal section 12–15 μm wide at sides. Conidiophores absent. Conidiogenous cells phialidic, 7–17 × 3.5–5 μm, doliiform to lageniform, formed all around the locular cavity. Conidia cylindrical, aseptate, hyaline, smooth, (9–)11.5–18(–21.5) × 3–4.5 μm (av. 14.4 × 3.4 μm, n = 31).

Specimen examined: Japan, Hokkaido, Isl. Rebun, Kafuka, Nairo-river, on dead stems of polygonaceous plant, 30 Aug. 2001, K. Tanaka (holotype KT 731 = HHUF 27558, culture ex-type CBS 139701 = JCM 19412 = MAFF 239468).

Notes: Additional details of this species were reported by Tanaka & Harada (2003b), who misidentified this fungus as Massarina rubi based mostly on the morphology of the sexual morph. However, Pseudocoleophoma is phylogenetically different from M. rubi, which belongs to the Lophiotremataceae (Zhang et al., 2009b, Hirayama and Tanaka, 2011b). This species resembles P. calamagrostidis but has larger ascospores (20.6 × 4.8 μm vs. 17.4 × 3.8 μm) and conidia (14.4 × 3.4 μm vs. 8.6 × 2.2 μm).

Didymosphaeriaceae Munk, Dansk Bot. Ark. 15 (2): 128. 1953.

= Montagnulaceae M.E. Barr, Mycotaxon 77: 194. 2001.

Type genus: Didymosphaeria Fuckel.

Karstenula Speg., Decades Mycologicae Italicae 7–12: no. 94. 1879.

Type species: Karstenula rhodostoma (Alb. & Schwein.) Speg.

Notes: This genus is phylogenetically close to Paraphaeosphaeria, but can be distinguished from the latter by the ascomata surrounded by well-developed subiculum and cylindrical asci with uniseriate ascospores.

Karstenula rhodostoma (Alb. & Schwein.) Speg., Decades Mycologicae Italicae 7–12: no. 94. 1879. Fig. 6.

Fig. 6.

Karstenula rhodostoma. A. Ascoma in longitudinal section; B. Ascomatal wall; C. Pseudoparaphyses; D. Ascus; E, F. Ascospores. All from UPS (F-141152) 425947. Scale bars: A = 50 μm; B–F = 10 μm.

Basionym: Sphaeria rhodostoma Alb. & Schwein., Consp. Fung. (Leipzig): 43. 1805.

Asci cylindrical, 145–175 × 12.5–14.5 μm (av. 162.2 × 13.4 μm, n = 20), with a short stipe (14–23 μm long). Ascospores cylindrical to ellipsoid, 22–26 × 8–10 μm (av. 23.7 × 9.1 μm, n = 20), l/w 2.3–2.9 (av. 2.6, n = 20), with a submedian primary septum (0.50–0.54; av. 0.52, n = 20), with 3 transverse septa (1+1+1), with or without 1 longitudinal septum.

Specimen examined: Sweden, “Odlingen” field, ca. 250 m S of Jerusalem, on twigs of Frangula alnus, Jan. 1995, K. Holm & L. Holm, UPS (F-141152) 425947, culture CBS 691.94.

Notes: For further description of this species including its asexual morph (Microdiplodia frangulae), see Constantinescu (1993) and Zhang et al. (2012). More than 360 species are recorded as Microdiplodia (MycoBank, http://www.mycobank.org), but the validity of this genus is unknown due to lack of phylogenetic information regarding the lectotype species, M. conigena (Clements & Shear 1931).

Neokalmusia Kaz. Tanaka et al., Fungal Divers. 68: 92. 2014.

Type species: Neokalmusia brevispora (Nagas. & Y. Otani) Kaz. Tanaka et al.

Notes: Neokalmusia is characterised by subglobose to oblong ascomata including several pseudothecia in a row and verrucose ascospores (Ariyawansa et al. 2014). Two bambusicolous species, N. brevispora on Sasa and N. scabrispora on Phyllostachys are known in this genus. Although they share many morphological features, monophyly of Neokalmusia was not supported in this study (Fig. 1). Additional taxa and sequence data of this genus are needed to evaluate validity of Neokalmusia.

Neokalmusia brevispora (Nagas. & Y. Otani) Kaz. Tanaka et al., Fungal Divers. 68: 92. 2014. Fig. 7.

Fig. 7.

Neokalmusia brevispora. A. Ascomata on the natural host surface; B, C. Ascomata in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath; I. in India ink); J. Germinating ascospore. A, F, G, J from KT 2313; B–D, I from KT 1466; E, H from culture KT 1466. Scale bars: A = 1 mm; B, C = 100 μm; D–J = 10 μm.

Basionym: Phaeosphaeria arundinacea var. brevispora Nagas. & Y. Otani, Rep. Tottori Mycol. Inst. 15: 38. 1977.

Specimens examined: Japan, Hokkaido, Sapporo, Moiwa-yama, on culms of Sasa sp., 13 Jun. 1972, E. Nagasawa (holotype of Phaeosphaeria arundinacea var. brevispora TMI 3175); Fukushima, Minamiaizu, Ose pond, on dead twigs of Sasa sp., 30 Aug. 2003, N. Asama, KT 1466 = HHUF 28229, culture CBS 120248 = JCM 13543 = MAFF 239276; Hokkaido, Isl. Rishiri, Afutoromanai-river, on dead twigs of Sasa kurilensis, 29 Jul. 2007, K. Tanaka & G. Sato (epitype designated here KT 2313 = HHUF 30016, MBT202863, culture ex-epitype NBRC 106240).

Notes: For other descriptions of this species, see Nagasawa and Otani, 1977, Shoemaker and Babcock, 1989, Tanaka and Harada, 2004, and Ariyawansa et al. (2014). Although this species was transferred to Kalmusia (Zhang et al. 2009b), subsequent molecular studies (Hyde et al., 2013, Zhang et al., 2014a) did not support this placement, and thus Neokalmusia was established to accommodate N. brevispora as the type species of this genus (Ariyawansa et al. 2014). We here designate an epitype specimen for this species.

Neokalmusia scabrispora (Teng) Kaz. Tanaka et al., Fungal Divers. 68: 92. 2014. Fig. 8.

Fig. 8.

Neokalmusia scabrispora. A. Ascomata on the natural host surface; B, C. Ascomata in longitudinal section (B. in lactophenol cotton blue); D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G, H. Ascospores (arrowheads indicate gelatinous sheath; H. in India ink); I. Germinating ascospores; J. Spermatia. A–D from KT 1023; E–I from KT 2202; J from culture KT 1023. Scale bars: A = 1 mm; B, C = 100 μm; D–J = 10 μm.

Basionym: Leptosphaeria scabrispora Teng, Sinensia, Shanghai 4: 378. 1934.

Specimens examined: Japan, Tochigi, Kanuma, near Ooashi-river, on dead twigs of Phyllostachys bambusoides, 6 Mar. 2003, N. Asama, KT 1023 = HHUF 28608, culture CBS 120246 = JCM 12851 = MAFF 239517; Kagoshima, Kumagegun, Isl. Yakushima, Miyanoura-river, riverbank, on dead twigs of Phyllostachys bambusoides, 17 Mar. 2007, K. Tanaka & H. Yonezawa, KT 2202 = HHUF 30013, culture NBRC 106237.

Notes: For a description of this species, see Tanaka et al. (2005a). This fungus was originally described as a Leptosphaeria (Teng 1934), and later transferred to Massariosphaeria (Shoemaker & Babcock 1989) or Kalmusia (Tanaka et al. 2005a). Molecular studies, however, did not support these placements (Tanaka et al., 2009, Zhang et al., 2014a). This species is currently treated as Neokalmusia (Ariyawansa et al. 2014), but we were not able to confirm the congenericity of N. scabrispora with N. brevispora (type species of Neokalmusia) (Fig. 1).

Paraphaeosphaeria O.E. Erikss., Ark. Bot. 6: 405. 1967.

Type species: Paraphaeosphaeria michotii (Westend.) O.E. Erikss.

Notes: The genus Paraphaeosphaeria was established by Eriksson (1967) as a generic segregate from Leptosphaeria. To date, about 35 taxa have been described as species within Paraphaeosphaeria (http://www.indexfungorum.org, Aug. 2015), of which nine species have been transferred to Neophaeosphaeria (Coniothyriaceae or Leptosphaeriaceae) or Phaeosphaeriopsis (Phaeosphaeriaceae) based on ascospore and conidial morphology, as well as sequence data from SSU and ITS nrDNA (Câmara et al., 2001, Câmara et al., 2003), and 10 asexual species with coniothyrium-like morphology have been added to this genus based on molecular evidence (Trakunyingcharoen et al., 2014, Verkley et al., 2014, Liu et al., 2015).

Paraphaeosphaeria michotii (Westend.) O.E. Erikss., Arkiv Bot. 6: 405. 1967. Fig. 9.

Fig. 9.

Paraphaeosphaeria michotii. A. Ascomata on the natural host surface; B. Ascoma in longitudinal section; C. Ascomatal wall; D. Ascus; E–H. Ascospores (arrowheads indicate gelatinous sheath; H. in India ink); I. Conidioma in culture (on rice straw); J. Conidioma in longitudinal section; K, L. Conidia. A–C from KT 2222; D–L from culture KT 2222. Scale bars: A, I = 500 μm; B, J = 50 μm; C–H, K, L = 10 μm.

Basionym: Sphaeria michotii Westend., Bull. Acad. R. Sci. Belg., Cl. Sci., sér. 2, 7: 87. 1859.

In culture, both sexual and asexual morphs were observed. Asci 77–90 × 13–14.5 μm. Ascospores ellipsoid, 18–24 × 5–7 μm (av. 21.3 × 6.2 μm, n = 30), l/w 2.8–3.8 (av. 3.5, n = 30), with a submedian primary septum (0.56–0.67; av. 0.61, n = 30), 2-septate (1+1+0). Conidia broadly fusiform to ellipsoid, 8–14.5 × 4–7 μm (av. 10.8 × 5.3 μm, n = 50), l/w 1.6–2.9 (av. 2.0, n = 50), echinulate.

Specimen examined: Japan, Tochigi, Utsunomiya, Ootanikannon, on dead leaves of Typha latifolia, 29 Mar. 2007, K. Tanaka & Y. Harada, KT 2222 = HHUF 30142, culture MAFF 243861.

Notes: Morphological features of the sexual morph in our material agree well with the description of P. michotii reported by several authors (Eriksson, 1967, Shoemaker and Eriksson, 1967, Shoemaker and Babcock, 1985, Ariyawansa et al., 2014). The conidia we observed are somewhat larger than those reported by Webster (1955; 6–10 × 3–5 μm) and Câmara et al. (2001; 4–8 × 2.4–4.4 μm), but match those reported by Sivanesan (1984; 5–13.5 × 3–5 μm). A BLAST search using ITS sequences showed 99.8 % similarity to sequences from the ex-epitype of P. michotii (GenBank KJ939279; 519/520) deposited in GenBank by Ariyawansa et al. (2014).

Spegazzinia Sacc., Michelia 2 (6): 37. 1880.

Type species: Spegazzinia ornata Sacc.

Notes: Based on the morphological features of basauxic conidiogenesis, Hyde et al. (1998) considered Spegazzinia to be a possible member of the Apiosporaceae (Sordariomycetes), although this was not fully resolved by molecular evidence (Hyde et al., 2011, Crous and Groenewald, 2013). We have assigned this genus to the Didymosphaeriaceae (Fig. 1).

Spegazzinia deightonii (S. Hughes) Subram., J. Indian Bot. Soc. 35: 78. 1956. Fig. 10.

Fig. 10.

Spegazzinia deightonii. A, B. Sporodochia on the natural host surface; C, D. Conidiophores and conidiogenous cells (arrowheads indicate basauxic conidiogenesis); E. Conidia (arrowheads indicate a type conidia, arrows indicate b type conidia). A from yone 66; B–E from culture yone 66. Scale bars: A, B = 500 μm; C–E = 10 μm.

Basionym: Spegazzinia tessarthra var. deightonii S. Hughes, Mycol. Pap. 50: 65. 1953.

Specimens examined: Japan, Kagoshima, Isl. Yakushima, Kurio, on dead leaves of Arundo donax, 14 Mar. 2007, K. Tanaka & H. Yonezawa, yone 66 = HHUF 30150, culture MAFF 243876; Okinawa, Isl. Iriomote, Inamori path, on dead stems of herbaceous plant, 25 Sep. 2007, H. Yonezawa & K. Tanaka, yone 212 = HHUF 30151, culture MAFF 243877.

Note: The above collections match the original description and illustration of S. tessarthra var. deightonii provided by Hughes (1953b).

Spegazzinia tessarthra (Berk. & M.A. Curtis) Sacc., Syll. Fung. 4: 758. 1886. Fig. 11.

Fig. 11.

Spegazzinia tessarthra. A. Sporodochia on the natural host surface; B, C. Conidiophores and conidiogenous cells (arrowheads indicate basauxic conidiogenesis); D. Conidia (arrowheads indicate a type conidia, arrows indicate b type conidia). All from SH 287. Scale bars: A = 500 μm; B–D = 10 μm.

Basionym: Sporidesmium tessarthrum Berk. & M.A. Curtis, J. Linn. Soc., Bot. 10: 355. 1868 (1869).

Specimen examined: Japan, Aomori, Towada, Sanbongi, Yagami, on balsa wood, 9 Nov. 2002, S. Hatakeyama, SH 287 = HHUF 27691, culture MAFF 243875.

Notes: The features of the collection match the description of S. tessarthra (Ellis 1971). The ITS sequences from the above isolate were identical to those of S. tessarthra in GenBank (GenBank JQ673429).

Lentitheciaceae Y. Zhang ter et al., Stud. Mycol. 64: 93. 2009.

Type genus: Lentithecium K.D. Hyde et al.

Katumotoa Kaz. Tanaka & Y. Harada, Mycoscience 46: 313. 2005.

Type species: Katumotoa bambusicola Kaz. Tanaka & Y. Harada.

Notes: Two bambusicolous genera, Katumotoa (Tanaka & Harada 2005) and Neoophiosphaerella, formed a clade as reported in previous molecular studies (Schoch et al., 2009, Tanaka et al., 2009, Zhang et al., 2012), but these have ascospores with distinct features, i.e., apiosporous in Katumotoa and multi-septate scolecosporous in Neoophiosphaerella. Furthermore, sequence similarity between the type species of both genera was 82.7 % (417/504) in their ITS regions, suggesting that they are not congeneric. Although a correlation between phylogenetic relationships and host preferences has been noted in these bambusicolous fungi (Zhang et al. 2012), discovery of additional species in both monotypic genera will be needed to confirm this.

Katumotoa bambusicola Kaz. Tanaka & Y. Harada, Mycoscience 46: 313. 2005. Fig. 12.

Fig. 12.

Katumotoa bambusicola. A. Ascomata on the natural host surface; B, C. Ascomata in longitudinal section; D. Ascomatal wall (in lactophenol cotton blue); E. Pseudoparaphyses; F. Ascus; G. Ascospore (arrowheads indicate gelatinous sheath). A–D from KT 1517; E–G from culture KT 1517. Scale bars: A = 1 mm; B, C = 100 μm; D–G = 10 μm.

Specimen examined: Japan, Iwate, Nishine, Hirakasa, near Yakebashiri, Mt. Iwate, on Sasa kurilensis, 19 Oct. 2003, K. Tanaka (paratype KT 1517a = HHUF 28661, culture ex-paratype JCM 13131 = MAFF 239641).

Keissleriella Höhn., Sber. Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1 128: 582. 1919.

Type species: Keissleriella aesculi (Höhn.) Höhn.

Notes: Keissleriella is characterised by an ostiolar neck covered by short dark setae. Munk (1953) introduced Trichometasphaeria, which can be separated from Keissleriella by their host preferences and the morphological features of hamathecium and ascospores. Trichometasphaeria occurs on herbaceous plants and has cellular pseudoparaphyses and septate ascospores, while Keissleriella occurs on woody plants and has trabecular pseudoparaphyses and 1-septate ascospores (Munk, 1957, Barr, 1990a, Barr, 1992). However, Bose (1961) treated Trichometasphaeria as a synonym of Keissleriella, and this opinion was accepted by several authors (e.g., Eriksson, 1967, Dennis, 1978, Sivanesan, 1984). We also regard these two genera as congeneric as species with the features of Trichometasphaeria (e.g., K. gloeospora, the type species of Trichometasphaeria) and Keissleriella (e.g., K. cladophila; Corbaz, 1956, Bose, 1961) form a clade (Fig. 1), although the type of the latter genus (K. sambucina) is not included in our analysis.

Keissleriella breviasca Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811304. Fig. 13.

Fig. 13.

Keissleriella breviasca. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Apical setae of ascoma; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–K. Ascospores (arrowheads indicate gelatinous sheath); L, M. Ascomata and conidiomata in culture (on rice straw); N. Ascoma produced in culture; O. Conidioma produced in culture; P. Conidia. A–C, E, I from KT 649; D, G, H from KT 540; F, J from KT 581; K from culture KT 581; L–O from culture KT 649; P from culture KT 540. Scale bars: A, L = 2 mm; B, M = 500 μm; C, N, O = 20 μm; D–K, P = 10 μm.

Etymology: From the Latin brevi- meaning short, in reference to the ascus length.

Ascomata scattered, erumpent, 95–115 μm high, 130–165 μm diam. Ostiolar neck papillate, 18–23(–50) μm long, covered with 1-celled, thick-walled, dark brown to almost black setae. Ascomatal wall 7.5–11.5 μm thick at sides, composed of 3–4 layers of polygonal to subglobose cells of 2.5–10 × 2.5–5 μm. Pseudoparaphyses cellular, septate, branched and anastomosed, 2–3.5 μm wide. Asci fissitunicate, clavate to cylindrical, 51–66 × 7–11 μm (av. 59.0 × 8.7 μm, n = 39), rounded at the apex and with a shallow ocular chamber, short-stalked (5–12 μm long), with 4 biseriate ascospores. Ascospores narrowly fusiform, 17.5–24.5 × 3.5–5 μm (av. 21.1 × 4.3 μm, n = 64), l/w (3.7–)4.0–5.7(–6.3) (av. 4.9, n = 64), with a nearly median primary septum (0.44–0.53; av. 0.49, n = 48), 3-septate (1+1+1), slightly constricted at the septa, hyaline, smooth, with an entire sheath; sheath gelatinous and 8–11 μm wide when fresh, delimited and 1–2 μm wide when dry.

Colonies on PDA (after 4 wk) attaining a diam of 1.0–1.3 cm, coral with white margin; reverse red to flesh; sienna pigment produced. In culture coelomycetous asexual morph formed. Conidiomata pycnidial, 70–90 μm high, 120–200 μm diam, subglobose to hemispherical in section. Conidiomatal wall 7–15 μm thick at sides, composed of thin-walled, flattened cells. Conidiogenous cells cylindrical to doliiform, 8–13 × 2.5–3.5 μm, holoblastic. Conidia cylindrical to bone-shaped, 0–3-septate, hyaline, smooth, 11–20 × 3–4 μm (av. 15.2 × 3.7 μm, n = 20), l/w 3.7–5.0 (av. 4.1, n = 20), without sheath. Sometimes, sexual morph formed. Ascospores 19–23 × 3.5–4.5 μm.

Specimens examined: Japan, Aomori, Hirosaki, Campus of Hirosaki Univ., on dead culms of Dactylis glomerata, 13 Jun. 2001, K. Tanaka, KT 540 = HHUF 27715, culture JCM 19413 = MAFF 239476; ibid., 20 Jul. 2001, KT 581 = HHUF 27717, culture JCM 19414 = MAFF 243843; Aomori, Hirakawa, Hirakawa-river, riverbank, on dead culms of Dactylis glomerata, 5 Aug. 2001, K. Tanaka (holotype KT 649 = HHUF 27718, culture ex-type CBS 139691 = JCM 19415 = MAFF 243844).

Notes: To date, 50 species have been described as Keissleriella (or Trichometasphaeria; MycoBank, http://www.mycobank.org, April 2015), but K. breviasca can be distinguished from all known species by its short asci with consistently four ascospores (Fig. 13G). The asexual morph of K. breviasca is similar to that of K. gallica reported as “Ascochyta sp.” (Sivanesan 1984), but the conidia of K. breviasca have 0–3 septa (vs. 0–1 septa in K. gallica). A collection of K. culmifida with 4-spored asci has sometimes been reported (Dennis, 1978, Ridley, 1988), which is most probably conspecific with K. breviasca.

Keissleriella culmifida (P. Karst.) S.K. Bose, Phytopath. Z. 41: 188. 1961. Fig. 14.

Fig. 14.

Keissleriella culmifida. A. Ascomata on the natural host surface; B. Ascomata in culture (on rice straw); C. Ascoma in longitudinal section; D. Apical setae of ascoma; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath); K. Germinating ascospore. A, C, E, K from KT 2642; B, D, F, G, I, J from culture KT 2308; H from KT 2308. Scale bars: A, B = 500 μm; C, D, K = 20 μm; E–J = 10 μm.

Basionym: Leptosphaeria culmifida P. Karst., Bidr. Känn. Finl. Nat. Folk 23: 103. 1873.

Ascomata 120–145 μm high, 120–150 μm diam, with dark brown setae of 30–58 × 3–5 μm around the ostiole. Asci 8-spored, 70.5–84 × 10.5–13 μm. Ascospores fusiform, 17.5–22 × 4–5.5 μm (av. 19.4 × 4.9 μm, n = 36), l/w 3.5–5.0 (av. 3.9, n = 36), with nearly median primary septum (0.47–0.53; av. 0.51, n = 35), 3-septate (1+1+1), surrounded by an entire gelatinous sheath, 3–9 μm wide.

In culture sexual morph formed. Asci 90–127.5 × 10.5–15 μm (av. 102.9 × 13.0 μm, n = 40). Ascospores fusiform, 19–26 × 5–7 μm (av. 22.5 × 5.9 μm, n = 60), l/w 3.3–4.5 (av. 3.8, n = 60), with a nearly median primary septum (0.46–0.55; av. 0.50, n = 60). Asexual morph not observed.

Specimens examined: Japan, Hokkaido, Isl. Rishiri, Forest park, on dead stems of Agrostis flaccida, 29 Jul. 2007, K. Tanaka & G. Sato, KT 2308 = HHUF 30135, culture JCM 19416 = MAFF 243848; Iwate, Hachimantai, Top of Aspite line, on dead leaves of Festuca sp. 25 Jul. 2009, K. Tanaka & Y. Harada, KT 2642 = HHUF 30136, culture JCM 19417 = MAFF 243849.

Notes: We identified these specimens as K. culmifida, based on the description (Karsten, 1873, Holm, 1957, Bose, 1961) and illustration (Berlese, 1894, Eriksson, 1967) of this species. However, ITS sequences from the above materials completely matched with those from the ex-type of K. poagena (GenBank KJ869112), a species recently published by Crous et al. (2014a). Keissleriella poagena is morphologically close to K. culmifida, and thus taxonomic reassessment of these two species will be needed.

Keissleriella gloeospora (Berk. & Curr.) S.K. Bose, Phytopath. Z. 41: 190. 1961. Fig. 15.

Fig. 15.

Keissleriella gloeospora. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Apical setae of ascoma; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath); K. Spermatia. A–J from KT 829; K from culture KT 829. Scale bars: A = 1 mm; B = 500 μm; C = 50 μm; D–K = 10 μm.

Basionym: Sphaeria gloeospora Berk. & Curr., Ann. Mag. Nat. Hist., Ser. 3 7: 454. 1861.

Ascomata 130–230 μm high, 350–480 μm diam, with dark brown setae (up to 60 μm long) around the ostiole. Asci clavate, 80–118 × 14–19 μm (av. 97.2 × 16.8 μm, n = 20), short-stalked (10–25 μm long). Ascospores fusiform to clavate, 21.5–33 × 6–9 μm (av. 28.0 × 7.3 μm, n = 50), l/w 3.3–4.9 (av. 3.8, n = 50), with a supramedian primary septum (0.39–0.48; av. 0.43, n = 50), 4–6-septate (1+1+2, 1+1+3, 2+1+2, 2+1+3), sometimes with a vertical septum in central cells, surrounded by an entire sheath (6–10 μm wide when fresh, 2–3 μm wide when dry).

In culture spermatial morph formed. Spermatia 3.3–4.9 × 1.2–1.8 μm (av. 4.1 × 1.5 μm, n = 20), hyaline, cylindrical.

Specimen examined: Japan, Aomori, Hirosaki, Campus of Hirosaki Univ., on dead culms of Setaria faberii, 4 Nov. 2001, K. Tanaka, KT 829 = HHUF 27704, culture MAFF 239474.

Note: The above material matches well the descriptions of K. gloeospora provided by Bose (1961) and Shearer et al. (1993).

Keissleriella quadriseptata Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811305. Fig. 16.

Fig. 16.

Keissleriella quadriseptata. A, B. Ascomata on the natural host surface; C, D. Ascomata in longitudinal section; E. Apical setae of ascoma; F. Ascomatal wall; G. Pseudoparaphyses; H. Ascus; I–K. Ascospores (arrowheads indicate gelatinous sheath); L. Germinating ascospore; M. Conidiomata in culture (on rice straw); N. Conidioma in longitudinal section; O. Conidiomatal wall; P. Conidium; Q. Senescent conidium. A–L from KT 2292; M–Q from culture KT 2292. Scale bars: A, M = 500 μm; B = 200 μm; C, D, N = 50 μm; E–L, O–Q = 10 μm.

Etymology: Referring to the 4-septate ascospores.

Ascomata scattered to 2–4 grouped, immersed to erumpent, globose in section, 210–320 μm high, 170–310 μm diam. Ostiolar neck central, papillate, 35–50 μm long, 55–62 μm diam, with setae; setae bluntly pointed, aseptate, slightly waved, dark brown to black, 37–93 μm long, 2.5–4 μm wide at the base. Ascomatal wall in longitudinal section uniformly 15–20 μm thick, composed of 5–6 layers of polygonal, slightly thick-walled, 4.5–16.5 × 3–7.5 μm, brown cells. Pseudoparaphyses cellular, septate, branched and anastomosed, 2–2.5 μm wide. Asci fissitunicate, clavate, 78.5–107.5 × 12–14.5 μm (av. 92.8 × 13.1 μm, n = 30), rounded at the apex and with a shallow ocular chamber, short-stalked (5–24 μm long; av. 13.1 μm, n = 26), with 8 biseriate ascospores. Ascospores clavate, 19–24.5 × 5–7 μm (av. 21.5 × 5.8 μm, n = 50), l/w 3.3–4.4 (av. 3.7, n = 50), with a supramedian primary septum (0.38–0.45; av. 0.42, n = 50), 4-septate (1+1+2), slightly constricted at the primary septum, hyaline, smooth, with an entire gelatinous sheath of 4–6 μm wide.

Colonies on PDA (after 2 wk) attaining a diam of 1.7–2.2 cm, pale luteous; reverse rust; no pigment produced. In culture asexual morph formed. Conidiomata pycnidial, 200–380 μm high, 160–310 μm diam, globose in section. Ostiolar neck papillate, 60–90 μm long, 50–75 μm wide. Conidiomatal wall 13–25 μm thick, composed of 4–6 layers of polygonal cells (5–17 × 2.5–6.5 μm). Conidiophores absent. Conidiogenous cells lageniform, holoblastic. Conidia cylindrical, rounded at the apex, slightly truncate at the base, straight, aseptate, hyaline, smooth, 25–32 × 6–8.5 μm (av. 28.4 × 7.2 μm, n = 30), l/w 3.3–4.8 (av. 3.9, n = 30), without sheath; senescent spores brown, 0–1-septate.

Specimen examined: Japan, Hokkaido, Isl. Rishiri, Beach near Ryuzinnoiwa, on dead culms of Dactylis glomerata, 28 Jul. 2007, K. Tanaka & G. Sato (holotype KT 2292 = HHUF 30137, culture ex-type CBS 139692 = JCM 19418 = MAFF 243850).

Notes: This species is characterised by its consistently 4-septate ascospores (1+1+2). Phylogenetically, it is closest to K. gloeospora (97.8 %; 904/924 in tef1), but the latter has larger ascospores (21.5–33 × 6–9 μm) with 4–6 septa (1+1+2, 1+1+3, 2+1+2, 2+1+3). In culture, K. quadriseptata produced an asexual morph with cylindrical, aseptate, hyaline conidia, but only a spermatial morph was found in K. gloeospora.

Keissleriella taminensis (H. Wegelin) S.K. Bose, Phytopath. Z. 41: 190. 1961. Fig. 17.

Fig. 17.

Keissleriella taminensis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Apical setae of ascoma; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath); K. Spermatia. A–E, G, I from KT 594; F, H from KT 571; J from KT 678; K from culture KT 594. Scale bars: A = 2 mm; B = 500 μm; C = 50 μm; D–K = 10 μm.

Basionym: Leptosphaeria taminensis H. Wegelin, Mitt. Thürgau. Naturf. Ges. 12: 173. 1896.

Ascomata 190–280 μm high, 180–300 μm diam. Asci clavate, 70–106 × 11.5–16 μm (av. 84.3 × 13.3 μm, n = 53), short-stalked (8–25 μm long; av. 15.5 μm, n = 27). Ascospores fusiform, 19–25 × 4–7 μm (av. 21.9 × 5.9 μm, n = 103), l/w 3.0–4.5(–5.0) (av. 3.7, n = 103), with a supramedian primary septum (0.43–0.51; av. 0.47, n = 100), 4–5-septate (1+1+2, 2+1+2), surrounded by an entire sheath (3–10 μm wide when fresh, 1–2 μm wide when dry).

In culture spermatial morph formed. Spermatia cylindrical, hyaline, 3.5–8.8 × 1.3–2.3 μm (av. 5.5 × 1.8 μm, n = 60). Sometimes sexual morph observed.

Specimens examined: Japan, Aomori, Hirosaki, Sanpinai, on dead stems of herbaceous plant, 17 Jul. 2001, K. Tanaka, KT 571 = HHUF 27707, culture MAFF 243846; Aomori, Hirosaki, Kadoke, Oowasawa-river, riverbank, on dead stems of herbaceous plant, 29 Jul. 2001, K. Tanaka, KT 594 = HHUF 27709, culture MAFF 243847; ibid., 14 Aug. 2001, K. Tanaka, KT 678 = HHUF 27711, culture MAFF 239475.

Note: These materials agree well with the description of K. taminensis except for the slightly wider ascospores in our collections (4–7 μm vs. 4.5–5 μm; Bose 1961).

Keissleriella yonaguniensis Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811307. Fig. 18.

Fig. 18.

Keissleriella yonaguniensis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Apical setae of ascoma; E. Ascomatal wall; F. Pseudoparaphyses; G. Asci; H–K. Ascospores (arrowheads indicate gelatinous sheath; K. in India ink). All from KT 2604. Scale bars: A = 2 mm; B = 500 μm; C = 50 μm; D–K = 10 μm.

Etymology: Referring to the collection site.

Ascomata scattered, immersed to erumpent, globose in section, 100–170 μm high, 100–180 μm diam. Ostiolar neck central, papillate to terete, 37–45 μm long, 32–35 μm diam, with setae; setae bluntly pointed, aseptate, straight, dark brown to black, 20–30 μm long, 3–4.5 μm wide at the base. Ascomatal wall in longitudinal section uniformly 7.5–10 μm thick, composed of 3–5 layers of polygonal, thin-walled, flattened, 3.5–7.5 × 1.5–2.5 μm, brown cells. Pseudoparaphyses cellular, septate, branched and anastomosed, 1.5–2.5 μm wide (3–4.5 μm wide at below). Asci fissitunicate, cylindrical, 65–99.5(–112.5) × 10.5–14.5 μm (av. 80.1 × 12.1 μm, n = 35), rounded at apex and with a shallow ocular chamber, with a short stipe of 5–13 μm long, 8-spored. Ascospores cylindrical with rounded ends, 15–20 × 4.5–6.5 μm (av. 18.1 × 5.3 μm, n = 50), l/w 3.0–3.9 (av. 3.5, n = 50), with a nearly median primary septum (0.47–0.53; av. 0.50, n = 50), 5-septate (2+1+2), yellow, smooth, with an entire gelatinous sheath of 3–5 μm wide (later diffuse up to 10 μm wide).

Colonies on PDA (after 4 wk) attaining a diam of 5.2–5.7 cm, white to pale luteous; reverse similar; no pigment produced. In culture sexual morph identical to that formed on the natural host produced.

Specimen examined: Japan, Okinawa, Isl. Yonaguni, Kubura pond, on dead leaves of Typha latifolia, 23 Nov. 2008, K. Tanaka & K. Hirayama (holotype KT 2604 = HHUF 30138, culture ex-type CBS 139693 = JCM 19419 = MAFF 243851).

Notes: Among the 50 described species in Keissleriella (or Trichometasphaeria), six are known to have 5-septate ascospores like K. yonaguniensis. These are K. abruptipapilla, K. gloeospora, T. papillisetosa, K. pindaundeensis, T. populi and K. taminensis. Among these, K. gloeospora, T. populi and K. taminensis differ from K. yonaguniensis in having asymmetrically-septate ascospores (Bose, 1961, Barr, 1992). Keissleriella pindaundeensis (Kobayasi 1971) and T. papillisetosa (Yuan & Barr 1994) are distinguished from K. yonaguniensis by larger ascospores that are more than 20 μm long. Keissleriella abruptipapilla (Barr 1990a) is similar but its ascospores are verruculose. In our phylogenetic tree (Fig. 1), K. yonaguniensis grouped with K. linearis (= Lentithecium lineare; Zhang et al. 2009c), Murilentithecium with muriform ascospores and conidia (Wanasinghe et al. 2014), and Phragmocamarosporium spp. with phragmosporous conidia (Wijayawardene et al. 2015), although this clade received no support. We include this species as Keissleriella based on the morphology of its sexual morph pending further studies of related taxa.

Lentithecium K.D. Hyde et al., Fungal Divers. 38: 234. 2009.

Type species: Lentithecium fluviatile (Aptroot & Van Ryck.) K.D. Hyde et al.

Notes: Lentithecium was established by Zhang et al. (2009c) using L. fluviatile as the type species. This genus was characterised by the lenticular ascomata, but reexamination based on the holotype of L. fluviatile revealed that the species has globose ascomata (Hyde et al. 2013).

Lentithecium clioninum (Kaz. Tanaka et al.) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811308. Fig. 19.

Fig. 19.

Lentithecium clioninum. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath; H, I. in Black-Blue ink). A–D from KT 1149A; E–G from KT 1220; H, I from culture KT 1149A. Scale bars: A = 1 mm; B = 500 μm; C = 50 μm; D–I = 10 μm.

Basionym: Massarina clionina Kaz. Tanaka et al., Mycoscience 46: 288. 2005.

Ascomata 210–280 μm high, 330–430 μm diam. Ostiolar neck 50–75 μm long, 75–125 μm wide. Ascomatal wall 15–23 μm thick at sides. Pseudoparaphyses 2 μm wide. Asci clavate, (81.5–)86–118(–128) × 15–19(–21) μm (av. 100.1 × 17.2 μm, n = 70), short-stalked (5–23 μm long). Ascospores fusiform, (26–)27.5–34.5(–37) × 7–10(–11) μm (av. 31.0 × 8.7 μm, n = 70), l/w 3.2–4.1 (av. 3.6, n = 70), with a septum mostly median (0.48–0.52; av. 0.50, n = 64), with a wing-like sheath staining with Black-Blue ink when fresh.

In culture ascomatal morph formed. Ascospores 27–35 × 8–11 μm (av. 31.0 × 9.3 μm, n = 50). No asexual morph observed.

Specimens examined: Japan, Hokkaido, Akkeshi, Bekanbeushi-river (near Bekanbeushi bridge), on submerged twigs of woody plant, 2 Jun. 2003, K. Tanaka & S. Hatakeyama (holotype KT 1149A = HHUF 28199, culture ex-type CBS 139694 = JCM 12703 = MAFF 239293); Hokkaido, Akkeshi, Toraibetsu-river, on submerged twigs of woody plant, 3 Jun. 2003, K. Tanaka & S. Hatakeyama (paratype KT 1220 = HHUF 28213, culture ex-paratype MAFF 243839).

Notes: This species was previously described as Massarina (Tanaka et al. 2005b), but we here transfer it to Lentithecium, because it has morphological similarities with L. fluviatile, the type species of the genus (see Fig. 43 in Zhang et al. 2012). Both have globose ascomata composed of small polygonal peridial cells, short pedicellate asci, and fusiform ascospores with obtuse ends. In our phylogenetic tree (Fig. 1), this species and L. pseudoclioninum nested on a well-supported branch (100 %) with L. fluviatile.

Lentithecium pseudoclioninum Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811309. Fig. 20.

Fig. 20.

Lentithecium pseudoclioninum. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath; all in Black-Blue ink); J. Germinating ascospore. All from KT 1113. Scale bars: A = 1 mm; B = 500 μm; C = 50 μm; D–J = 10 μm.

Etymology: Referring to its resemblance to Lentithecium clioninum.

Ascomata scattered to 2–3 grouped, immersed to erumpent, globose in section, 200–220 μm diam, with sparse brown hyphae around ascomata. Ostiolar neck central, papillate, 40–50 μm long, 30 μm diam, with periphyses. Ascomatal wall in longitudinal section uniformly 17.5–20 μm thick, composed of 5–6 layers of polygonal to subglobose, 5–12.5 × 2 μm, brown cells. Pseudoparaphyses septate, branched and anastomosed, 2–3 μm wide. Asci fissitunicate, clavate, 62.5–116 × 14–25 μm (av. 92.0 × 18.4 μm, n = 95), rounded at the apex and with a shallow ocular chamber, short-stalked (5–17.5 μm long; av. 9.9 μm, n = 67), with 8 biseriate ascospores. Ascospores clavate to broadly fusiform, slightly curved, 22–39 × 6.5–11.5 μm (av. 29.2 × 8.5 μm, n = 77), l/w (2.4–)2.8–4.1(–4.6) (av. 3.5, n = 77), with a supramedian septum (0.43–0.50; av. 0.48, n = 73), constricted at the septum, hyaline, smooth, with an amorphous gelatinous sheath (1–4 μm wide) staining with Black-Blue ink when in fresh condition.

Colonies on PDA (after 4 wk) attaining a diam of 2.7–3.0 cm, smoke grey; reverse vinaceous buff to citrine; no pigment produced. In culture sexual morph identical to that formed on the natural host produced.

Specimens examined: Japan, Aomori, Hirosaki, Aoki, Mohei pond, on submerged twigs of woody plant, 3 May 2003, K. Tanaka & N. Asama, KT 1111 = HHUF 29053, culture JCM 19421 = MAFF 243840; ibid. (holotype KT 1113 = HHUF 29055, culture ex-type CBS 139695 = JCM 19422 = MAFF 243841).

Notes: Morphologically, this species is close to L. clioninum, but is clearly separated from it on the basis of its smaller ascospores with a supramedian septum. The wing-like sheath of ascospores found in L. clioninum was not observed in L. pseudoclioninum.

Neoophiosphaerella Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811310.

Etymology: After its morphological similarity to Ophiosphaerella.

Ascomata scattered to grouped, erumpent, subglobose to hemispherical with flattened base. Ostiolar neck central, terete, flush, covered by black clypeus. Ascomatal wall at sides composed of several layers of thin-walled brown cells. Pseudoparaphyses numerous, cellular, branched. Asci fissitunicate, cylindrical, short-stalked, with 8 parallel or twisted ascospores. Ascospores filiform, multiseptate, hyaline to pale yellowish brown, smooth, surrounded by a sheath. Asexual morph unknown.

Type species: Neoophiosphaerella sasicola (Nagas. & Y. Otani) Kaz. Tanaka & K. Hiray.

Notes: Neoophiosphaerella sasicola, the type species of this genus, was originally described as a species of Phaeosphaeria (Nagasawa & Otani 1977) and later transferred to Ophiosphaerella (Shoemaker & Babcock 1989). These two genera, however, belong to the Phaeosphaeriaceae in the Pleosporineae (Câmara et al., 2000, Schoch et al., 2009, Phookamsak et al., 2014), a family distantly related to the Lentitheciaceae. We therefore propose a new genus, Neoophiosphaerella, to accommodate this species. Ophiosphaerella has globose to subglobose ascomata with a papillate ostiolar neck (Phookamsak et al. 2014), while Neoophiosphaerella is characterised by hemispherical ascomata without papilla but being covered by clypei.

Neoophiosphaerella sasicola (Nagas. & Y. Otani) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811312. Fig. 21.

Fig. 21.

Neoophiosphaerella sasicola. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G. Ascospore (arrowheads indicate gelatinous sheath; in India ink). All from KT 1706. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–G = 10 μm.

Basionym: Phaeosphaeria sasicola Nagas. & Y. Otani, Rep. Tottori Mycol. Inst. 15: 39. 1977.

Specimens examined: Japan, Hokkaido, Ebestu, Nopporo, on Sasa senanensis, 15 May 1972, E. Nagasawa (holotype of Phaeosphaeria sasicola TMI 3176); Hokkaido, Isl. Rebun, Funadomari, Akaiwa, on dead culms of Sasa kurilensis, 5 Jun. 2004, K. Tanaka (epitype designated here KT 1706 = HHUF 29443, MBT202864, culture ex-epitype CBS 120247 = JCM 13134 = MAFF 239644).

Note: The collection HHUF 29443 is designated as epitype for N. sasicola, the type species of Neoophiosphaerella.

Setoseptoria Quaedvl. et al., Stud. Mycol. 75: 382, 2013.

Type species: Setoseptoria phragmitis Quaedvl. et al.

Notes: Quaedvlieg et al. (2013) established the genus Setoseptoria typified by S. phragmitis on Phragmites. The sexual morph of this coelomycetous genus is presently unknown.

Setoseptoria arundinacea (Sowerby) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811313. Fig. 22.

Fig. 22.

Setoseptoria arundinacea. A. Surface view of ascomata; B, C. Ascomata in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G, H. Ascospores (arrowheads indicate gelatinous sheath). All from KT 600. Scale bars: A = 500 μm; B = 100 μm; C = 50 μm; D–H = 10 μm.

Basionym: Sphaeria arundinacea Sowerby, Col. Fig. Engl. Fung. 3: 139, t. 336. 1803.

Specimens examined: Japan, Aomori, Hirosaki, Sanpinai, on dead culms of Phragmites australis, 1 Jul. 2001, K. Tanaka, KT 552 = HHUF 27543, culture MAFF 239460; Aomori, Hirosaki, Kadoke, Oowasawa-river, on dead culms of Phragmites australis, 29 Jul. 2001, K. Tanaka, KT 600 = HHUF 27544, culture MAFF 243842.

Notes: This species has been placed in various pleosporalean genera, such as Leptosphaeria (see Crane & Shearer 1991), Lophiostoma (Hyde et al. 2002), Massarina (Leuchtmann 1984), Metasphaeria (Vasilyeva 1998), and Phaeosphaeria (Hedjaroude 1968). Most recently, it has been transferred to Lentithecium based on the results of phylogenetic analyses using SSU + LSU nrDNA and rpb2 (Zhang et al. 2009c). However, later molecular studies (Schoch et al., 2009, Shearer et al., 2009, Zhang et al., 2009b, Zhang et al., 2012, Liu et al., 2011), as well as our own work (Fig. 1), do not support this placement. This species and its phenotypically and phylogenetically close relative Massarina magniarundinacea (Tanaka et al. 2004) do not belong to any genera previously suggested. They should, therefore, be transferred to another genus. One candidate genus to accommodate these species would be Setoseptoria. The monotypic genus Setoseptoria typified by S. phragmitis was introduced for a stagonospora-like pycnidial coelomycete with (1–)3-septate, subcylindrical, hyaline conidia (Quaedvlieg et al. 2013). The sexual morph of Setoseptoria is unknown. In contrast, two massarina-like species, M. arundinacea and M. magniarundinacea, have been reported to produce only sexual morphs in culture (Lucas, 1968, Leuchtmann, 1984, Tanaka and Harada, 2003b, Tanaka et al., 2004). Although there is no example of production of stagonospora-like asexual morphs from the massarina-like sexual morphs in culture, congeneric relationships have been suggested several times between Stagonospora elegans (Aptroot, 1998, Eriksson and Hawksworth, 2003) or S. vexata (Grove 1935) and M. arundinacea, based on their close association on the same host tissue. Furthermore, two stagonospora-like species (Setoseptoria phragmitis and “Stagonospora” macropycnidia) and two massarina-like species (M. arundinacea and M. magniarundinacea) form a strongly supported clade (Fig. 1) and sequence similarities between these species in the LSU region are considerably higher (826/834 = 99.0 %). We therefore tentatively assign the two massarina-like species to the genus Setosphaeria, although asexual morphs of these species are presently unknown. A species with both sexual and asexual morphs will be required to confirm the validity of our generic treatment.

The two isolates of S. arundinacea that we examined did not form a clade with S. arundinacea from GenBank (Fig. 1). This species has been reported many times as a common species on Phragmites culms (Aptroot, 1998, Tanaka and Harada, 2003b), but S. arundinacea may consist of several cryptic species with close morphological resemblance.

Setoseptoria magniarundinacea (Kaz. Tanaka & Y. Harada) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811314. Fig. 23.

Fig. 23.

Setoseptoria magniarundinacea. A, B. Surface view of ascomata; C. Ascoma in longitudinal section (in lactophenol cotton blue); D. Ascomatal wall (in lactophenol cotton blue); E. Pseudoparaphyses; F. Ascus; G, H. Ascospores (arrowheads indicate gelatinous sheath); I. Germinating ascospore. A, B, E–H from culture KT 1174; C, D, I from KT 1174. Scale bars: A = 1 mm; B = 500 μm; C = 50 μm; D–I = 10 μm.

Basionym: Massarina magniarundinacea Kaz. Tanaka & Y. Harada, Mycotaxon 90: 349. 2004.

Ascomata subglobose to hemispherical with flattened base, 150–280 μm high, 310–410 μm diam. Ostiolar neck central, short papillate. Ascomatal wall 12.5–20 μm thick at sides. Pseudoparaphyses cellular, 1.5–3.5 μm wide. Asci (119–)125–182.5(–200) × 25–35(–47.5) μm (av.151.5 × 30.2 μm, n = 37). Ascospores 67–82 × 6.5–9 μm (av. 74.0 × 7.7, n = 50), l/w 8.4–11.0 (av. 9.6, n = 50), with a submedian primary septum (0.52–0.57; av. 0.54, n = 50).

In culture the ascomatal morph is similar to that observed on the natural host.

Specimen examined: Japan, Hokkaido, Akkeshi, Ariake, small stream, on submerged stems of herbaceous plant, 3 Jun. 2003, K. Tanaka & S. Hatakeyama (holotype KT 1174 = HHUF 28293, culture ex-type CBS 139702 = MAFF 239294).

Note: The ascospores of S. magniarundinacea are most similar in shape and colour to those of S. arundinacea, but are considerably larger (67–82 × 6.5–9 μm vs. 23–40 × 3.5–6 μm, Tanaka et al. 2004).

Tingoldiago K. Hiray. & Kaz. Tanaka, Mycologia 102: 740. 2010.

Type species: Tingoldiago graminicola K. Hiray. & Kaz. Tanaka.

Notes: Tingoldiago, found in freshwater environments, is characterised by lens-shaped ascomata and narrowly fusiform ascospores, each of which has an elongated sheath (Hirayama et al. 2010; Fig. 27). This genus, however, was regarded as a synonym of Lentithecium, despite the fact that the Lentithecium clade that included Tingoldiago was not well-supported (21 %; Zhang et al. 2012). When Lentithecium was established, the lenticular ascomata with simple peridial structure and hyaline 1-septate ascospores were emphasised as important characters to define the genus (Zhang et al. 2009c), but this generic circumscription is incorrect (see Notes in Lentithecium).

Fig. 27.

Stagonospora perfecta. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall in surface view; E. Ascomatal wall at side; F. Pseudoparaphyses; G. Asci; H. Apex of ascus; I–K. Ascospores (arrowheads indicate gelatinous sheath; I. in Black-Blue ink; K. in India ink); L, M. Conidiomata in culture (on rice straw); N, O. Conidiogenous cells (arrowheads indicate annellations); P. Conidioma in longitudinal section; Q. Conidia. A–K from KT 1726; L–Q from culture KT 1726. Scale bars: A, L = 1 mm; B, M = 200 μm; C, P = 20 μm; D–K, N, O, Q = 10 μm.

Tingoldiago graminicola K. Hiray. & Kaz. Tanaka, Mycologia 102: 740. 2010. Fig. 24.

Fig. 24.

Tingoldiago graminicola. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G. Ascospore (arrowheads indicate gelatinous sheath; in India ink). A–D, F, G from KH 68; E from culture KT 891. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–G = 10 μm.

Specimens examined: Japan, Hokkaido, Isl. Rishiri, Himenuma (pond), on submerged culms of Phragmites australis, 27 Jul. 2007, K. Hirayama & K. Tanaka (holotype KH 68 = HHUF 30009, culture ex-type JCM 16485 = NBRC 106131); ibid., 25 Jul. 2008, K. Hirayama & K. Tanaka (paratype KH 155 = HHUF 30010, culture ex-paratype JCM 16486 = NBRC 106132); Aomori, Hirosaki, Kadoke, Oowasawa-river, on submerged culms of Phragmites japonica, 28 Sep. 2002, K. Tanaka (paratype KT 891 = HHUF 27882, culture ex-paratype MAFF 239472).

Notes: The morphological features of this species have been described by Hirayama et al. (2010). Tingoldiago graminicola and Setoseptoria arundinacea, formerly treated as Lentithecium by Zhang et al., 2009c, Zhang et al., 2012, have lens-shaped ascomata, but are not in the clade of Lentithecium s. str. (Fig. 1). Instead, Lentithecium is restricted to species with globose ascomata (Fig. 19, Fig. 20).

Massarinaceae Munk, Friesia 5: 305. 1956.

Type genus: Massarina Sacc.

Helminthosporium Link, Mag. Gesell. naturf. Freunde, Berlin 3(1–2): 10. 1809.

Type species: Helminthosporium velutinum Link.

Notes: Although more than 700 taxa have been described as species within Helminthosporium (http://www.indexfungorum.org, Aug. 2015), the genus has been quite heterogeneous. Several unrelated pathogens of the Poaceae were segregated from Helminthosporium to other genera, i.e., Bipolaris (= Cochliobolus), Curvularia (= Pseudocochliobolus), Exserohilum (= Setosphaeria), and Pyrenophora (= Drechslera), all of which belong to the Pleosporaceae (Sivanesan, 1987, Hyde et al., 2013). Further distantly related species (e.g., H. asterinum) in the Leotiomycetes were excluded from Helminthosporium, and this genus was restricted to species having phylogenetic affinity with the Massarinaceae (Olivier et al., 2000, Kodsueb et al., 2007, Hyde et al., 2013).

Little is known about the sexual morphs of Helminthosporium s. str. Hughes (1953a) reported that an ascospore isolate of a Massaria species found on Quercus produced a Helminthosporium asexual morph. Subramanian & Sekar (1987) described Splanchnonema kalakadense as the sexual morph of H. velutinum based on cultural study. However, the validity of the generic classification based on the sexual morphs of these two examples remains unknown. The sexual morph of H. massarinum differs from those of Massaria (Massariaceae; Voglmayr & Jaklitsch 2011) and Splanchnonema (Pleomassariaceae; Hyde et al. 2013) in its ellipsoidal hyaline ascospores, and is rather similar to that of Massarina, although M. eburnea (the type species of Massarina) does not have a hyphomycetous asexual morph like that of Helminthosporium.

Helminthosporium massarinum Kaz. Tanaka, K. Hiray. & Shirouzu, sp. nov. MycoBank MB811315. Fig. 25.

Fig. 25.

Helminthosporium massarinum. A, B. Ascomata on the natural host surface; C, D. Ascomata in longitudinal section (in lactophenol cotton blue); E. Ascomatal wall (in lactophenol cotton blue); F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath; J. in Black-Blue ink); K. Germinating ascospore; L–O. Conidiophores; P. Conidiogenous cells (arrows indicate tretic pores); Q–S. Conidia; T. Spermogonium; U. Spermatia. A–I, K from KT 1564; J from KT 838; L–U from culture KT 838. Scale bars: A = 2 mm; B, L, M, T = 200 μm; C, D, N, O = 50 μm; E–K, P–S, U = 10 μm.

Etymology: Referring to the similarity of the sexual morph with that of the genus Massarina.

Ascomata numerous, scattered to 2–4 grouped, immersed below the host epidermis, hemispherical to subglobose with somewhat flattened base, 315–390 μm high, 300–430 μm diam. Ostiolar neck central, cylindrical to papillate, 125–165 μm long, 75–125 μm wide, composed of subglobose, 3–5 μm diam, brown cells, surrounded by dark brown clypeus-like structure, without periphyses. Ascomatal wall surface of textura prismatica, in a longitudinal section 12–18 μm wide at sides and base, composed of 5–7 layers of polygonal to rectangular, 5–15 × 2.5–6.5 μm, brown cells. Pseudoparaphyses cellular, 1.5–3 μm wide, septate at 7–16 μm long intervals, branched, anastomosed. Asci fissitunicate, clavate, 82–135 × 13–16 μm (av. 111.0 × 14.8 μm, n = 90), rounded at the apex, with a narrow apical chamber and faint ring, short-stalked (7–24 μm long; av. 16.7 μm, n = 35), with 8 ascospores biseriate above and uniseriate below. Ascospores ellipsoidal with rounded ends, mostly straight, 20–25(–27) × 5–8 μm (av. 22.6 × 7.0 μm, n = 100), l/w 2.7–3.9 (av. 3.2, n = 100), with a submedian septum (0.50–0.58; av. 0.54, n = 100), constricted at the septum, asymmetric, with wider upper cell, hyaline, guttulate, smooth, with a conspicuous gelatinous sheath of 2–3 μm wide in fresh condition (with a delimited firm sheath of 1 μm wide in dry condition).

Colonies on PDA (after 4 wk) attaining a diam of 5.5–7.1 cm, white to olivaceous grey; reverse smoke grey to buff; no pigment produced. In culture hyphomycetous asexual morph and spermatial morph formed. Conidiophores macronematous, mononematous, 380–810 μm long (av. 587 μm, n = 20), 7–9 μm wide at the apex, 13.5–21 μm wide at the base, arising singly or in groups of 4–5 from the stromata, straight or slightly curved, dark brown, guttulate, 15–25-septate at 8–45 μm long intervals, with small (ca. 1–2 μm diam) pores at the apex and beneath of septa, with thick wall of 1–4 μm wide. Conidia tretic, solitary or in short chains (5–6), 1–8-septate, 17–56.5 × 5–9 μm (av. 37.9 × 7.4 μm, n = 95), l/w 2.9–7.5 (av. 5.1, n = 95) [but mostly 4–6-septate, 27–53 × 6–9 μm (av. 38.7 × 7.4 μm, n = 64), l/w 4.1–6.8 (av. 5.3, n = 64)], obclavate, rostrate, pale brown, smooth, with or without guttules. Conidia germinating from both end cells. Spermogonia pycnidial, produced under the conidiophores, solitary or gregarious, 100–150(–300) μm diam, subglobose, uniloculate to multiloculate. Spermatiogenous cells cylindrical, up to 5 μm long, appearing phialidic. Spermatia 3–4(–5) × 2–2.5 μm (av. 3.5 × 2.2 μm, n = 26), globose to subglobose, hyaline, smooth.

Specimens examined: Japan, Aomori, Towada, Sanbongi, Yagami, on vines of Berchemia racemosa, 15 Dec. 2001, S. Hatakeyama, KT 838 = HHUF 27573, culture JCM 13094 = MAFF 239604; ibid., 23 Nov. 2003, S. Hatakeyama (holotype KT 1564 = HHUF 29089, culture ex-type CBS 139690 = JCM 13095 = MAFF 239605); ibid., KT 1565 = HHUF 29090; ibid., KT 1566 = HHUF 29091; ibid., 2 Dec. 2003, K. Tanaka, S. Hatakeyama & N. Nakagawara, KT 1613–1615 = HHUF 29092–29094.

Note: This species is somewhat similar to H. hypselodelphyos in having solitary or catenate conidia but the latter has smaller conidia (15–28 × 6.5–8 μm, av. 25 × 7.1 μm) (Ellis 1961).

Massarina Sacc., Syll. Fung. 2: 153. 1883.

Type species: Massarina eburnea (Tul. & C. Tul.) Sacc.

Notes: Since establishment of the genus (Saccardo 1883), more than 176 taxa have been described within Massarina (Index Fungorum; http://www.indexfungorum.org, Aug. 2015), but the heterogeneity of the genus has been continuously suggested. Several taxonomic reassessments of many species within Massarina have been attempted based on morphology (Bose, 1961, Barr, 1992, Hyde, 1995, Aptroot, 1998, Hyde et al., 2002, Tanaka and Harada, 2003b). It has more recently been revealed that the genus is highly polyphyletic based on molecular data (Liew et al. 2002), and most species in Massarina except for the type (M. eburnea) have been excluded from the genus. The genera segregated from Massarina s. lat. are phylogenetically diverse groups in the Pleosporales; Halomassarina (Trematosphaeriaceae; Suetrong et al. 2009), Lentithecium and Tingoldiago (Lentitheciaceae; Zhang et al., 2009c, Hirayama et al., 2010), Lindgomyces (Lindgomycetaceae; Hirayama et al. 2010), Morosphaeria (Morosphaeriaceae; Suetrong et al. 2009), and Triplosphaeria (Tetraplosphaeriaceae; Tanaka et al. 2009).

Massarina eburnea (Tul. & C. Tul.) Sacc., Syll. Fung. 2: 153. 1883. Fig. 26.

Fig. 26.

Massarina eburnea. A, B. Ascomata on the natural host surface; C. Side view of ascoma; D. Ascoma in longitudinal section; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath); K. Spermogonia in culture; L. Spermatia. A–J from H 3953; K, L from culture H 3953. Scale bars: A = 1 mm; B, C, K = 500 μm; D = 100 μm; E–J, L = 10 μm.

Basionym: Massaria eburnea Tul. & C. Tul., Select. Fung. Carpol. 2: 239. 1863.

Ascomata scattered to 2–4 grouped, immersed, hemispherical with flattened base or depressed globose, 300–420 μm high, 570–680 μm diam. Ostiolar neck short papillate, 60–90 μm long, central, with black clypeus (250–400 μm wide). Ascomatal wall 12–18 μm thick at sides, composed of 3–5 layers of thin-walled prismatic cells (5–12.5 × 2–4 μm). Pseudoparaphyses numerous, cellular, 2–3(–4.5) μm wide. Asci fissitunicate, clavate, (110–)120–180 × 21.5–30 μm (av. 141.6 × 25.6 μm, n = 50), with a stipe of 12.5–25 μm long (av. 18.6 μm, n = 38). Ascospores broadly fusiform with rounded ends, 34–40 × 12–15(–16) μm (av. 37.2 × 13.9 μm, n = 50; including firm sheath of 1–2 μm thick), l/w 2.5–2.9 (av. 2.7, n = 50), 3-septate (primary septum submedian: 0.51–0.55, av. 0.53, n = 50), hyaline, smooth, with a sharply delimited firm sheath of 1–2 μm thick (but up to 6 μm thick when fresh).

Specimen examined: UK, Wales, Swansea, dead twigs of Fagus sylvatica, Apr. 2001, H 3953 = HHUF 26621, culture CBS 139697 = JCM 14422.

Notes: The morphological characteristics of the above specimen fit well with those in the description of M. eburnea (Hyde 1995). A pycnidial morph of M. eburnea reported as Ceratophoma sp. (Bose, 1961, Sivanesan, 1984) was observed in our isolate, but the “conidia” did not germinate on several agar media (e.g., water agar, PDA, CMA, and MEA) over 7 d. The Ceratophoma morph is therefore considered to be spermatial in function.

Stagonospora (Sacc.) Sacc., Syll. Fung. 3: 445. 1884.

Type species: Stagonospora paludosa (Sacc. & Speg.) Sacc.

Notes: Stagonospora has been defined morphologically based on a broad generic concept (Sutton 1980) and has been believed to have phylogenetic affinities with the Phaeosphaeriaceae (Zhang et al. 2012). However, in a comprehensive phylogenetic study on Stagonospora and morphologically similar genera, Quaedvlieg et al. (2013) revealed that Stagonospora is polyphyletic and Stagonospora s. str. (based on S. paludosa) belongs to the Massarinaceae. Several new genera, such as Neostagonospora and Parastagonospora, have been established to accommodate unrelated stagonospora-like species in the Phaeosphaeriaceae (Quaedvlieg et al. 2013).

Neottiosporina, typified by N. apoda (Subramanian 1961) may have phylogenetic relationships with Stagonospora, although phylogenetic placement of the type species has not been clarified. At least N. paspali, transferred from Stagonospora to Neottiosporina (Sutton & Alcorn 1974), should be regarded within Stagonospora, because this taxon has close morphological and phylogenetic affinity to Stagonospora species (Fig. 1).

Stagonospora bicolor (D. Hawksw. et al.) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811316.

Basionym: Leptosphaeria bicolor D. Hawksw. et al., Mycologia 71: 483. 1979.

≡ Saccharicola bicolor (D. Hawksw. et al.) D. Hawksw. & O.E. Erikss., Mycologia 95: 431. 2003.

Notes: Although we did not observe any material of this species, the illustration in the original description of Leptosphaeria bicolor (Kaiser et al. 1979), as well as SSU (GenBank U04202) and ITS (GenBank U04203) sequences derived from the type culture (ATCC 42652) of this species clearly indicates that it is a member of Stagonospora, as recently circumscribed by Quaedvlieg et al. (2013). Based on L. bicolor, Eriksson & Hawksworth (2003) erected Saccharicola for leptosphaeria-like species on sugarcane, but Saccharicola should be regarded as a synonym of Stagonospora. The sexual morph of Stagonospora s. str. has been referred to as didymella-like (Quaedvlieg et al. 2013), and some species of Didymella with a stagonospora-like asexual morph (e.g., D. proximella on Carex; Corlett & Smith 1978) may have phylogenetic affinity with Stagonospora s. str. based on their morphologies.

Stagonospora perfecta Quaedvlieg et al., Stud. Mycol. 75: 378. 2013. Fig. 27.

Ascomata scattered, immersed, globose to subglobose in section, 160–220 μm high, 140–250 μm diam. Ostiolar neck central, papillate to short cylindrical, 37–50 μm long, 52–75(–100) μm wide. Ascomatal wall in longitudinal section uniformly 12–20 μm thick, composed of 4–6 layers of polygonal to subglobose, brown cells of 5–20 × 3.5–10 μm. Pseudoparaphyses cellular, septate, branched and anastomosed, 2.5–4 μm wide. Asci fissitunicate, cylindrical, 75–115 × 15–20.5 μm (av. 89.8 × 17.5 μm, n = 30), rounded at the apex and with a shallow ocular chamber, short-stalked (7–14.5 μm long), with 8 biseriate ascospores. Ascospores broadly fusiform, 20–28 × 6.5–11 μm (av. 23.4 × 8.3 μm, n = 50), l/w 2.3–3.3 (av. 2.8, n = 50), with a submedian septum (0.54–0.59; av. 0.56, n = 50), constricted at the septum, hyaline, smooth, with an entire sheath; sheath gelatinous, 2–7 μm, staining with Black-Blue ink when in fresh condition, delimited and 1–3 μm wide when in dry condition.

In culture both sexual and asexual morphs formed. Conidiomata pycnidial, 80–100 μm high, 100–120 μm diam, scattered, globose in section. Conidiomatal wall in longitudinal section uniformly 6–13 μm wide, composed of 3–4 layers of polygonal, thin-walled, 7–17 × 2.5–6 μm, pale brown cells. Conidiophores absent. Conidiogenous cells annellidic, cylindrical to subglobose, 8–17 × 5–9 μm. Conidia cylindrical, 3(–5)-septate, hyaline, smooth, (25–)27–37(–48) × 8–11 μm (av. 32.9 × 9.5 μm, n = 60), l/w 2.7–4.1(–5.1) (av. 3.5, n = 60), without sheath. Ascospores in culture slightly larger than those on the natural host, 27–33 × 10–12 μm.

Specimen examined: Japan, Hokkaido, Isl. Rebun, Funadomari, Kusyuko (pond), on dead leaves of Carex sp., 3 Jun. 2004, K. Tanaka, KT 1726A = HHUF 29095, culture JCM 13099 = MAFF 239609.

Notes: The characteristics of the above material match those in the original description of S. perfecta described from Carex acutiformis in the Netherlands (Quaedvlieg et al. 2013), except that the conidia in our material were wider (8–11 μm vs. 6–8 μm). LSU and ITS sequences obtained from our material and those from the ex-type of S. perfecta (GenBank LSU: KF251761, ITS: KF251258) were identical. The sexual morph of this species has been referred to as didymella-like (Quaedvlieg et al. 2013), and fits well with the generic concept of Saccharicola (Eriksson & Hawksworth 2003), but this genus should be synonymised under the older genus Stagonospora (Saccardo 1884).

Stagonospora pseudoperfecta Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811317. Fig. 28.

Fig. 28.

Stagonospora pseudoperfecta. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath); J. Conidia. A–I from KT 889; J from culture KT 889. Scale bars: A = 2 mm; B = 500 μm; C = 20 μm; D–J = 10 μm.

Etymology: After its morphological similarity to Stagonospora perfecta.

Ascomata scattered, immersed, globose in section, 200–250 μm high, 210–260 μm diam. Ostiolar neck central, papillate, 35–63 μm long, 63 μm wide. Ascomatal wall in longitudinal section uniformly 10–15 μm thick, composed of 3–5 layers of polygonal to subglobose, brown cells of 6–16.5 × 3.5–11.5 μm. Pseudoparaphyses cellular, numerous, 3–6 μm wide, septate, branched, anastomosed. Asci fissitunicate, clavate, 66–100 × 13.5–17.5 μm (av. 86.3 × 15.4 μm, n = 20), rounded at apex and with a shallow apical chamber, short-stalked (7–17.5 μm long), with 8 irregularly biseriate to triseriate ascospores. Ascospores narrowly fusiform, straight or slightly curved, 21–30.5 × 5–7 μm (av. 26.0 × 5.7 μm, n = 40), l/w 3.3–5.9 (av. 4.6, n = 40), with submedian septum (0.50–0.57, av. 5.4, n = 40), slightly constricted at the septum, hyaline, with or without guttules, smooth, with an entire sheath; sheath gelatinous, 0.5–2 μm wide at side.

Colonies on PDA (after 2 wk) attaining a diam of 4.9–5.5 cm, white; reverse sepia to fuscous black; no pigment produced. In culture asexual morph formed. Conidiomata pycnidial. Conidia cylindrical, aseptate, hyaline, 21.5–26 × 4–5.5 μm (av. 24.1 × 4.9 μm, n = 10), l/w 4.6–5.8 (av. 5.0, n = 10).

Specimens examined: Japan, Aomori, Hirosaki, Kadoke, Oowasawa-river, riverbank (Horikoshi-bridge), on dead leaves of Typha latifolia, 8 Sep. 2002, K. Tanaka (holotype KT 889 = HHUF 29087, culture ex-type CBS 120236 = JCM 13097 = MAFF 239607); ibid., KT 888 = HHUF 29086.

Notes: Eight species are currently accepted in Stagonospora s. str. based on molecular evidence (Crous et al., 2013b, Crous et al., 2014b, Quaedvlieg et al., 2013). Stagonospora pseudoperfecta is similar to S. perfecta, but has slightly longer and more slender ascospores (l/w 4.6 vs. 2.8). In terms of overall morphology and host preference, S. pseudoperfecta superficially resembles “Massarina” lacustris sensu Leuchtmann (1984) (non Wettsteinina lacustris sensu Shoemaker & Babcock 1989, or Khashnobish & Shearer 1993) reported from Typha and Schoenoplectus. However, molecular data (SSU, rpb2, tef1) obtained from a strain studied by Leuchtmann (CBS 618.86) suggests that the latter fungus is a member of the Lentitheciaceae (Schoch et al. 2009). In ITS analysis with other Stagonospora species (data not shown), S. pseudoperfecta positioned as a sister taxon to S. duoseptata, but similarity between the two taxa in this region was 95.2 % (452/475) with 1.3 % (6/475) gaps.

In culture, pycnidial conidiomata with cylindrical hyaline conidia were observed only once, but this may have been an immature condition because the conidia were aseptate and smaller (Fig. 28J). Unfortunately, the asexual morph in culture failed to be observed again, despite several attempts.

Stagonospora tainanensis W.H. Hsieh, Mycologia 71: 893. 1979. Fig. 29.

Fig. 29.

Stagonospora tainanensis. A, B. Ascomata on the natural host surface; C. Ascomata and conidiomata in culture (on rice straw); D. Ascoma in longitudinal section; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath); K. Conidia. A, B, D, E, G, H from KT 1866; C, F, I–K from culture KT 1866. Scale bars: A, C = 2 mm; B = 500 μm; D = 20 μm; E–K = 10 μm.

Asci ovoid to cylindrical, 102–122.5 × 26.5–32.5 μm, 8-spored. Ascospores fusiform, 36–44 × 8.5–12 μm, l/w 3.6–4.7, with a submedian septum (0.53–0.56), hyaline, smooth, with an entire sheath (2–5 μm wide when fresh, 1–2 μm wide when dry).

In culture, both sexual and asexual morphs formed. Ascospores 32–40 × 10.5–13 μm, l/w 2.8–3.2, with a submedian septum (0.54–0.56). Conidia ellipsoid, 37–48(–55) × 12–13(–15) μm, l/w 3.0–3.8, 3-septate, hyaline.

Specimen examined: Japan, Kagoshima, Isl. Yakushima, Nunobikinotaki park, on dead leaves of herbaceous plant, 19 Oct. 2005, K. Tanaka & T. Hosoya, KT 1866 = HHUF 30141, culture MAFF 243860.

Notes: The ITS sequence from this material is identical with the sequence (GenBank AF439464) of Stagonospora taiwanensis obtained from the ex-type culture (ATCC 38204; Hsieh 1979), and two ITS sequences (GenBank AF439462, AF439463) of Saccharicola taiwanensis on Saccharum. An unnamed “Saccharicola” on Miscanthus (O'Neill & Farr 1996), a grass genus related to Saccharum, is also considered to be conspecific, because these have identical ITS sequences (GenBank AF439467; Câmara et al. 2002) and morphological similarities (O'Neill & Farr 1996). Morphologically, our specimen agrees with the previous description of this species (as Leptosphaeria taiwanensis; Hsieh, 1979, Sivanesan, 1984, Sivanesan and Waller, 1986), but the large ascospores reported by Shoemaker & Babcock (1989) were not observed.

Morosphaeriaceae Suetrong et al., Stud. Mycol. 64: 161. 2009.

Type genus: Morosphaeria Suetrong et al.

Aquilomyces D.G. Knapp et al., Persoonia 35: 93. 2015.

Type species: Aquilomyces patris D.G. Knapp et al.

Notes: Aquilomyces was erected by Knapp et al. (2015) to accommodate A. patris, a root endophyte of white poplar. There was no morphological information of this genus, because no fructifications were observed for A. patris.

Aquilomyces rebunensis Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811322. Fig. 30.

Fig. 30.

Aquilomyces rebunensis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section (in lactophenol cotton blue); D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–K. Ascospores (arrowheads indicate gelatinous sheath); L. Spermatia. A–D, F–I from KT 732; E, J–L from culture KT 732. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–L = 10 μm.

Etymology: After the locality where this fungus was collected.

Ascomata subglobose, 350–550 μm high, (430–)600–700 μm diam, covered with numerous brown hyphae of 2–3 μm thick. Ostiolar neck terete, central, 85–130 μm long, 75–130 μm wide, clypeate, with periphyses, composed of carbonaceous polygonal cells of 5–10 × 3–5 μm. Ascomatal wall at side wedge-shaped, up to 125 μm thick, composed of vertically orientated angular brown cells (5–8 × 2–5 μm); wall at the base 25–50 μm thick, composed of 3–8 μm diam, subglobose, brown cells. Pseudoparaphyses branched and anastomosed, septate, 1.5–2 μm wide. Asci fissitunicate, cylindrical to clavate, 97.5–147.5 × 15–21 μm (av. 121.3 × 18.3 μm, n = 45), with a short stipe of 7.5–25(–37.5) μm (av. 18.0 μm, n = 34). Ascospores fusiform, slightly curved, 30–38.5 × 6.5–11.5 μm (av. 35.1 × 8.6 μm, n = 50), l/w 3.3–5.0(–5.6) (av. 4.2, n = 50), with a supramedian septum (0.44–0.50; av. 0.47, n = 57), hyaline, smooth, with a sheath of 1–2 μm wide.

Colonies on PDA (after 4 wk) attaining a diam of 4.3–4.6 cm, pale olivaceous grey with white margin; reverse black to olivaceous grey; no pigment produced. In culture spermatial and ascomatal morphs formed. Spermatia 2–3 × 1.5–1.8 μm, subglobose to oblong. Ascospores slightly larger than those found on nature, 35–48 × 8–11 μm (av. 39.7 × 9.3 μm, n = 25).

Specimen examined: Japan, Hokkaido, Isl. Rebun, Kafuka, Nairo-river, on submerged twigs of woody plant, 30 Aug. 2001, K. Tanaka (holotype KT 732-2 = HHUF 27556, culture ex-type CBS 139684 = JCM 19427 = MAFF 243862).

Notes: We tentatively describe this aquatic fungus as Aquilomyces based on our molecular results. Phylogenetic analysis showed that A. patris, the type species of Aquilomyces, and A. rebunensis clustered into a distinct and moderately-supported clade (85 %, Fig. 1). Sequence similarities between these two taxa were 97.7 % (858/878) with 2.3 % gaps (20/878) in LSU and 95.4 % (576/604) with 0.7 % gaps (4/604) in ITS, suggesting that they are congeneric.

Clypeoloculus Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811318.

Etymology: Referring to the morphology of the ascomata covered by clypeus.

Ascomata scattered to 2–3 grouped, immersed to erumpent, subglobose or hemispherical with flattened base in section, covered with numerous brown hyphae around ascomata. Ostiolar neck terete to papillate, central, clypeate, composed of carbonaceous cells. Ascomatal wall wedge-shaped or “rim-like” at sides. Pseudoparaphyses septate, branched and anastomosed. Asci fissitunicate, clavate, with a short stipe, 8-spored. Ascospores narrowly fusiform, slightly curved, with a primary septum median to supramedian, constricted at the septum, hyaline, smooth, surrounded by an entire gelatinous sheath. Asexual morph unknown.

Type species: Clypeoloculus akitaensis Kaz. Tanaka & K. Hiray.

Notes: A new genus, Clypeoloculus, is proposed here for freshwater species having globose to subglobose ascomata with prominent clypeus, clavate asci, and 1-septate, hyaline ascospores with an entire sheath. These morphological characters fit with the broad generic concept of Massarina (Aptroot, 1998, Tanaka and Harada, 2003b), but Clypeoloculus can be distinguished from Massarina s. str. by the ascomata with wedge-shaped or “rim-like” ascomatal wall and narrowly fusiform ascospores. Furthermore, species in Clypeoloculus are located in the Morosphaeriaceae clade and are distantly related to M. eburnea, the type species of Massarina (Massarinaceae; Fig. 1).

Clypeoloculus akitaensis Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811319. Fig. 31.

Fig. 31.

Clypeoloculus akitaensis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath); J. Spermogonium in culture (on rice straw); K. Spermogonium in longitudinal section (in lactophenol cotton blue); L. Spermatia. A–I from KT 788; J–L from culture KT 788. Scale bars: A = 2 μm; B, J = 500 μm; C, K = 100 μm; D–I, L = 10 μm.

Etymology: Referring to the location where the specimen was collected.

Ascomata scattered to 2–3 grouped, immersed to erumpent, globose to subglobose in section, 400–550 μm high, 580–720 μm diam, covered with numerous brown hyphae around ascomata. Ostiolar neck terete, central, clypeate, composed of carbonaceous cells. Ascomatal wall in longitudinal section 37–100 μm wide at sides and base, composed of subglobose to globose, thick-walled, 2.5–10 μm diam, brown cells. Pseudoparaphyses septate, branched and anastomosed, 2–2.5 μm wide. Asci fissitunicate, clavate, (95–)110–155 × 17.5–27.5 μm (av. 125.1 × 20.9 μm, n = 30 μm), with a short stipe of (7.5–)12–27.5(–30) μm (av. 19.4 μm, n = 26) long, with 8 biseriate ascospores. Ascospores narrowly fusiform, slightly curved, 33.5–43 × 7–10 μm (av. 38.7 × 8.6 μm, n = 50), l/w 3.8–5.2 (av. 4.5, n = 50), with a primary septum supramedian (0.44–0.50; av. 0.48, n = 47), constricted at the septum, hyaline, smooth, surrounded by an entire gelatinous sheath, 1.5–3 μm wide.

Colonies on PDA (after 4 wk) attaining a diam of 2.0–2.2 cm, greenish grey; reverse black to smoke grey; no pigment produced. Spermatial morph observed in culture. Spermatia oblong, hyaline, 2.8–4 × 1.3–1.9 μm (av. 3.3 × 1.7 μm, n = 30).

Specimen examined: Japan, Akita, Kisakata, Akagawa-river, riverbank, on submerged twigs of woody plant, 23 Sep. 2001, K. Tanaka (holotype KT 788 = HHUF 27557, culture ex-type CBS 139681 = JCM 19424 = MAFF 239467).

Notes: In ascospore dimensions, C. akitaensis (av. 38.7 × 8.6 μm) is most similar to C. towadaensis (av. 38.7 × 8.7 μm), but the latter species has hemispherical ascomata with flattened bases and longer asci [(112–)120–170 μm long]. In the ITS sequences of these two species, there were differences at 29 positions.

Tanaka & Harada (2003b) reported this fungus as Massarina peerallyi (Hyde & Aptroot 1998). However, our re-examination of M. peerallyi based on its holotype [HKU (M) 2409] revealed that they are distinct species, because M. peerallyi has broader asci (80–127.5 × 25–35 μm) and larger ascospores (37–45 × 9–12 μm) with a supramedian septum (0.43–0.50 av. 0.48, n = 32). The morphological features of M. peerallyi are generally in accordance with the generic concept of Clypeoloculus, but phylogenetic reassessment using molecular sequences will be necessary before a new combination is proposed for this species.

Clypeoloculus hirosakiensis Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811320. Fig. 32.

Fig. 32.

Clypeoloculus hirosakiensis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath; I. in India ink); J. Germinating ascospore; K. Spermogonia in culture (on rice straw); L. Spermatia. A–J from KT 1283; K, L from culture KT 1283. Scale bars: A = 1 mm; B, K = 500 μm; C = 100 μm; D–J, L = 10 μm.

Etymology: Referring to the collection site.

Ascomata scattered, immersed to erumpent, globose to subglobose in section, 350–520 μm high, 340–550 μm diam, covered with numerous brown hyphae around ascomata. Ostiolar neck terete, central, 75–100 μm long, 40–170 μm wide, clypeate, composed of carbonaceous cells. Ascomatal wall in longitudinal section at sides, 50–60 μm wide, composed of parallel rows of rectangular to polygonal, brown to dark brown cells of 7.5–12.5 × 3.5–9.5 μm; wall at the base 17–25 μm wide, composed of subglobose to polygonal, slightly thick-walled, 3–10 × 3.5–6.5 μm, brown cells. Pseudoparaphyses septate, branched and anastomosed, 1.5–3 μm wide. Asci fissitunicate, clavate, 100–147.5 × (13.5–)15–20 μm (av. 118.6 × 16.5 μm, n = 50 μm), with a short stipe of (7.5–)10–25(–30) μm (av. 16.5 μm, n = 50) long, with 8 biseriate ascospores. Ascospores narrowly fusiform, slightly curved, (24–)29–36 × 7–9.5 μm (av. 33.0 × 8.0 μm, n = 50), l/w 3.7–4.7 (av. 4.1, n = 50), with a primary septum nearly median (0.47–0.52; av. 0.50, n = 50), constricted at the septum, hyaline, smooth, surrounded by an entire gelatinous sheath, 3–7 μm wide.

Colonies on PDA (after 4 wk) attaining a diam of 2.7–3.0 cm, greenish grey to lavender grey; reverse chestnut; ochreous pigment produced. In culture spermatial morph formed. Spermatia oblong, hyaline, 2.6–3.5 × 1.2–1.5.

Specimen examined: Japan, Aomori, Hirosaki, Zatoishi (river), on submerged twigs of woody plant, 19 Jul. 2003, K. Tanaka & N. Asama (holotype KT 1283 = HHUF 30144, culture ex-type CBS 139682 = JCM 19425 = MAFF 243864).

Note: The ascospores of C. hirosakiensis (av. 33.0 × 8.0 μm) are larger than those of C. microsporus (av. 29.9 × 7.0 μm) but smaller than those of C. akitaensis (av. 38.7 × 8.6 μm) and C. towadaensis (av. 38.7 × 8.7 μm).

Clypeoloculus microsporus Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811321. Fig. 33.

Fig. 33.

Clypeoloculus microsporus. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–J. Ascospores (arrowheads indicate gelatinous sheath); K. Spermogonium in culture (on rice straw); L. Spermatia. A–J from KT 1131; K, L from culture KT 1131. Scale bars: A = 1 mm; B, K = 500 μm; C = 100 μm; D–J, L = 10 μm.

Etymology: Referring to the small ascospores.

Ascomata scattered to 2–3 grouped, immersed to erumpent, globose to subglobose in section, 290–310 μm high, 280–350 μm diam, covered with sparse brown hyphae around ascomata. Ostiolar neck central, papillate, 70–90 μm long, 100–120 μm wide, clypeate, composed of carbonaceous cells. Ascomatal wall in longitudinal section 25–38 μm wide at sides and 25–30 μm wide at the base, composed of subglobose brown cells (2.5–10 μm diam). Pseudoparaphyses branched and anastomosed, septate, 2–3 μm wide. Asci fissitunicate, cylindrical to clavate, 80–145 × 13.5–22.5 μm (av. 115.5 × 17.5 μm, n = 33), with a short stipe of (5–)7.5–15(–29.5) μm (av. 13.9 μm, n = 30) long, with 8 biseriate to triseriate ascospores. Ascospores narrowly fusiform, slightly curved, 25.5–34.5 × 6–8(–9) μm (av. 29.9 × 7.0 μm, n = 56), l/w 3.4–5.2(–5.5) (av. 4.3, n = 56), with a primary septum nearly median (0.47–0.52; av. 0.50, n = 56), constricted at the septum, hyaline, smooth, surrounded by an entire gelatinous sheath of 2–4 μm wide.

Colonies on PDA (after 4 wk) attaining a diam of 3.1–3.3 cm, olivaceous grey; reverse umber to chestnut; amber pigment produced. In culture spermatial morph formed. Spermatia oblong, hyaline, 2.5–3.2 × 1.2–1.6 μm.

Specimen examined: Japan, Aomori, Kuroishi, Lake Nizinoko, on submerged twigs of woody plant, 3 May 2003, K. Tanaka & N. Asama (holotype KT 1131 = HHUF 30143, culture ex-type CBS 139683 = JCM 19426 = MAFF 243863).

Note: This species is distinguished from other species of Clypeoloculus by having the smallest ascospores known in the genus.

Clypeoloculus towadaensis Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811323. Fig. 34.

Fig. 34.

Clypeoloculus towadaensis. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–J. Ascospores (arrowheads indicate gelatinous sheath; J. senescent ascospore); K. Spermogonia in culture; L. Spermatia. A–J from KT 1340; K, L from culture KT 1340. Scale bars: A = 1 mm; B, K = 500 μm; C = 100 μm; D–J, L = 10 μm.

Etymology: Referring to the collection site.

Ascomata scattered to 2–3 grouped, immersed to erumpent, hemispherical with flattened base in section, 260–360 μm high, 550–700 μm diam. Ostiolar neck central, terete, 65–135 μm long, 75–88 μm wide, clypeate, composed of carbonaceous cells. Ascomatal wall at sides 95–150 μm wide and “rim-like”, composed of vertically orientated, rectangular to subglobose, hyaline to pale brown cells of 6–10 × 3–6 μm; at the base 15–25 μm wide or poorly developed. Pseudoparaphyses septate, branched and anastomosed, 1–2 μm wide. Asci fissitunicate, cylindrical, (112–)120–170 × 17.5–25 μm (av. 136.7 × 21.3 μm, n = 35), with a short stipe of (7.5–)12.5–25 μm (av. 17.2 μm, n = 35) long, with 8 biseriate ascospores. Ascospores narrowly fusiform, slightly curved, 34–43 × 7–10 μm (av. 38.7 × 8.7 μm, n = 50), l/w 3.9–5.3 (av. 4.5, n = 50), with a primary septum nearly median (0.47–0.53; av. 0.50, n = 50), constricted at the septum, hyaline, smooth, surrounded by an entire gelatinous sheath of 1–5 μm wide. Senescent ascospores becoming brown, 1–3(–5)-septate.

Colonies on PDA (after 4 wk) attaining a diam of 1.9–2.2 cm, greenish grey; reverse buff to sepia; ochreous pigment produced. In culture spermatial morph formed. Spermatia oblong to subglobose, hyaline, 3–4 × 1.4–1.8 μm.

Specimen examined: Japan, Akita, Kazuno, Kosaka, Lake Towada, on submerged twigs of woody plant, 26 Jul. 2003, K. Tanaka & N. Asama (holotype KT 1340 = HHUF 30145, culture ex-type CBS 139685 = JCM 19428 = MAFF 243865).

Notes: The most distinctive feature of C. towadaensis is its hemispherical ascomata with “rim-like” ascomatal walls. The asci of this species are longer than those of other species in Clypeoloculus.

Helicascus Kohlm., Canad. J. Bot. 47: 1471. 1969.

Type species: Helicascus kanaloanus Kohlm.

Notes: Helicascus was revised by Zhang et al. (2013) who accepted six species: two marine (H. kanaloanus and H. nypae) and four freshwater species (H. aegyptiacus, H. aquaticus, H. elaterascus and H. thalassioideus). Recently, two additional species in freshwater habitat (H. gallicus and H. unilocularis) were described in the genus (Zhang et al., 2014b, Zhang et al., 2015).

Helicascus aquaticus H. Zhang & K.D. Hyde, Sydowia 65: 155. 2013. Fig. 35.

Fig. 35.

Helicascus aquaticus. A, B. Ascomata on the natural host surface; C–E. Ascomata in longitudinal section; F. Ostiolar neck of ascoma; G. Ascomatal wall; H. Pseudoparaphyses; I. Ascus; J–L. Ascospores; M. Germinating ascospore; N. Spermogonium in culture (on rice straw); O. Spermatia. A–M from KT 1544; N, O from culture KT 1544. Scale bars: A = 1 mm; B, N = 500 μm; C–E = 100 μm; F, M = 20 μm; G–L, O = 10 μm.

Ascomata scattered to 2–3 grouped, immersed, hemispherical in section, 170–300 μm high (excluding the long ostiolar neck), 230–530 μm diam. Ostiolar neck cylindrical, oblique to central, 230–340 μm long, 110–160 μm wide, composed of polygonal to subglobose, thick-walled, 2.5–7.5 × 2–4 μm, brown to dark brown cells, with dense periphyses. Ascomatal wall in longitudinal section, 40–110 μm thick at sides, “rim-like”, composed of parallel rows of rectangular to polygonal, brown cells of 3.5–20 × 2.5–10 μm; at base poorly developed, 7–15 μm thick, composed of thin-walled flattened cells. Pseudoparaphyses septate, branched and anastomosed, 1.5–2(–3.5) μm wide, associated with gelatinous material. Asci fissitunicate, cylindrical to clavate, 80–122.5 × 15–19.5 μm (av. 102.8 × 17.5 μm, n = 20), with a stipe of 17.5–32.5 μm long (av. 24.7 μm long, n = 20), 8-spored. Ascospores broadly fusiform, slightly curved, 21.5–29 × 8–10.5 μm (av. 24.4 × 9.1 μm, n = 50), l/w 2.2–3.3 (av. 2.7, n = 50), with a submedian septum (0.51–0.60; av. 0.57, n = 50), constricted at the septum, pale greenish brown to brown, smooth, without sheath.

In culture spermatial morph formed. Spermatia oblong, hyaline, 2.4–4 × 1.5–2 μm.

Specimen examined: Japan, Aomori, Hirosaki, Aoki, Mohei-pond, on submerged twigs of woody plant, 25 Oct. 2003, K. Tanaka & N. Asama, KT 1544 = HHUF 30146, culture JCM 19423 = MAFF 243866.

Notes: The ITS sequence from the above isolate showed 100 % identity with that from the ex-type of H. aquaticus (GenBank KC886639), an aquatic species formerly known only from the type specimen collected on submerged wood in Thailand (Zhang et al. 2013). Our collection has a longer ascomatal neck (vs. 130–200 μm) and shorter asci [vs. 90–140(–185) μm] than described in the original publication. The ascospores possessing three septa were noted as the most distinctive feature of H. aquaticus (Zhang et al. 2013), but these were consistently 1-septate in our material.

Helicascus elaterascus (Shearer) H. Zhang & K.D. Hyde, Sydowia 65: 158. 2013. Fig. 36.

Fig. 36.

Helicascus elaterascus. A. Ascomata on the natural host surface; B. Ascomata in culture (on rice straw); C–E. Ascomata in longitudinal section; F. Ostiolar neck of ascoma; G. Ascomatal wall; H. Pseudoparaphyses; I. Ascus; J–L. Ascospores (arrowheads indicate gelatinous sheath); M. Germinating ascospore. A, C–G, I, M from KT 2673; B, H, J–L from culture KT 2673. Scale bars: A, B = 500 μm; C–E = 100 μm; F, M = 20 μm; G–L = 10 μm.

Basionym: Kirschsteiniothelia elaterascus Shearer, Mycologia 85: 963. 1994 (1993).

Ascomata scattered, immersed, depressed globose in section, 250–300 μm high (excluding the long ostiolar neck), 600–910 μm diam. Ostiolar neck 200–230 μm long, 110–160 μm wide, with dense periphyses. Pseudoparaphyses branched and anastomosed, septate, associated with gelatinous material. Asci fissitunicate, clavate, ca. 140 × 19 μm, with a long stipe, 8-spored. Ascospores broadly fusiform, slightly curved, 26.5–31.5 × 9–12.5 μm (av. 29.0 × 10.6 μm, n = 30), l/w 2.4–3.1 (av. 2.8, n = 30), with a submedian septum (0.50–0.58; av. 0.54, n = 30), slightly constricted at the septum, pale greenish brown to brown, with an entire sheath slightly enlarged below. In culture only sexual morph observed.

Specimens examined: Japan, Okinawa, Isl. Ishigaki, Mt. Omoto (river), on submerged twigs of woody plant, 2 Dec. 2009, Y. Kurihara, KT 2673 = HHUF 30147, culture MAFF 243867; Okinawa, Isl. Iriomote, Aira-river, on submerged twigs of woody plant, 8, Dec. 2009, Y. Kurihara & A. Ooba, KT 2682 = HHUF 30451, culture CBS 139689.

Notes: The morphological features of the examined material matched well with those in the original description of this species (Shearer 1993, as Kirschsteiniothelia elaterascus) and the LSU sequences were identical with that of the species deposited in GenBank (GenBank AY787934; Kodsueb et al. 2006). This species has been reported frequently from submerged wood in temperate and tropical freshwater habitats (Shearer and Raja, 2010, Raja et al., 2011) including Japan (Tsui et al. 2003).

Helicascus thalassioideus (K.D. Hyde & Aptroot) H. Zhang & K.D. Hyde, Sydowia 65: 159. 2013. Fig. 37.

Fig. 37.

Helicascus thalassioideus. A, B. Ascomata on the natural host surface; C. Ascomata in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G. Ascospore. All from KH 242. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–G = 10 μm.

Basionym: Massarina thalassioidea K.D. Hyde & Aptroot, Nova Hedwigia: 498. 1998.

Asci clavate, 80–120 × 15–17.5 μm (av. 100.2 × 16.7 μm, n = 30), with a stipe of 15–32.5 μm long (av. 23.0 μm, n = 30). Ascospores 25–31 × 8–10 μm (av. 27.5 × 8.6 μm, n = 30), l/w 2.8–3.5 (av. 3.2, n = 30), with a median septum. Asexual morph unknown.

Specimens examined: Japan, Okinawa, Isl. Iriomote, Geta-river, on submerged twigs of woody plant, 22 Nov. 2008, K. Hirayama & K. Tanaka, KH 242 = HHUF 30069, culture JCM 17526 = NBRC 107811. China, Hong Kong, Lam Tsuen river, N.T. on submerged wood, culture CBS 110441 = JCM 14147.

Notes: A more detailed description of this species was provided by Hirayama & Tanaka (2011a). This species was originally described as Massarina (Hyde & Aptroot 1998) and has commonly been reported from submerged wood in freshwater habitats throughout the world including Australia, Brunei, China (Yunnan), French West Indies, Hong Kong, the Philippines, and Thailand (Zhang et al., 2013, Zhang et al., 2015). Zhang et al. (2014b) suspected that helicascus-like taxa with a worldwide distribution might be a species-complex. However, ITS sequences obtained from the above two isolates were identical and differed by only one position (517/518) from that of H. thalassioideus collected in French West Indies (GenBank KP637162, Zhang et al. 2015).

Morosphaeria Suetrong et al., Stud. Mycol. 64: 161. 2009.

Type species: Morosphaeria velatispora (K.D. Hyde & Borse) Suetrong et al.

Notes: The genus Morosphaeria currently comprises two marine species, M. ramunculicola and M. velatispora (Suetrong et al. 2009). A freshwater fungus, Kirschsteiniothelia elaterascus, was transferred to Morosphaeria (Boonmee et al. 2012), but it is presently placed in Helicascus (Zhang et al. 2013).

Morosphaeria ramunculicola (K.D. Hyde) Suetrong et al., Stud. Mycol. 64: 162. 2009. Fig. 38.

Fig. 38.

Morosphaeria ramunculicola. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G. Ascospore (arrowheads indicate gelatinous sheath). All from KH 220. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–G = 10 μm.

Basionym: Massarina ramunculicola K.D. Hyde, Mycologia 83: 839. 1992 (1991).

Asci cylindrical to clavate, 121–187.5 × 22–32.5 μm, stalked (20–42.5 μm long). Ascospores 33–41 × 11–15 μm (av. 36.5 × 12.6 μm, n = 30), l/w 2.4–3.4 (av. 2.9, n = 30), with a submedian primary septum (0.50–0.56; av. 0.52, n = 30). Asexual morph unknown.

Specimen examined: Japan, Okinawa, Isl. Iriomote, Oomijya-river, on twigs of Rhizophora mucronata, 22 Nov. 2008, K. Hirayama & K. Tanaka, KH 220 = HHUF 30070, culture NBRC 107813.

Notes: The morphological features of the specimen cited here were identical to those of M. ramunculicola (Hyde 1991). There were only 1–2 bp differences between the LSU sequences (ca. 1 280 bp) of our material and those of M. ramunculicola deposited in GenBank (GenBank GQ925853, GQ925854).

Morosphaeria velatispora (K.D. Hyde & Borse) Suetrong et al., Stud. Mycol. 64: 161. 2009. Fig. 39.

Fig. 39.

Morosphaeria velatispora. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G. Ascospore (arrowheads indicate gelatinous sheath). A, D from KH 218; B, C, E–G from KH 221. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–G = 10 μm.

Basionym: Massarina velatispora K.D. Hyde & Borse, Mycotaxon 27: 161. 1986.

Asci clavate, (135–)165–245(–280) × 25–38 μm (av. 212.2 × 32.8 μm, n = 30), with a stipe of 25–55 μm long (av. 37.0 μm, n = 26). Ascospores 42–52 × 14–18 μm (av. 47.1 × 16.1 μm, n = 40), l/w 2.6–3.4 (av. 2.9, n = 40), 1–3-septate, with a median primary septum (0.49–0.53; av. 0.50, n = 40). Asexual morph unknown.

Specimens examined: Japan, Okinawa, Isl. Iriomote, Oomijya-river, on twigs of Rhizophora mucronata, 22 Nov. 2008, K. Hirayama & K. Tanaka, KH 218 = HHUF 30072, culture JCM 17529 = NBRC 107814; Okinawa, Isl. Iriomote, Geda river, on twigs of Rhizophora mucronata, 20 Nov. 2008, K. Hirayama & K. Tanaka, KH 221 = HHUF 30073, culture JCM 17530 = NBRC 107812.

Note: Hirayama & Tanaka (2011a) provided a more precise description based on the above specimens.

Parabambusicolaceae Kaz. Tanaka & K. Hiray., fam. nov. MycoBank MB811324.

Etymology: Referring to the name of the type genus.

Ascomata scattered to grouped, immersed to erumpent, globose to subglobose in surface view, depressed globose to hemispherical in section. Ostiolar neck or short papillate, central, sometimes compressed, composed of heavily melanised cells. Ascomatal wall composed of pale brown cells, with or without “rim-like” stromatic region. Pseudoparaphyses septate, branched and anastomosed. Asci fissitunicate, clavate to broadly cylindrical, stipitate, with 8 ascospores. Ascospores clavate to fusiform, with a supramedian primary septum, multiseptate, slightly constricted at septa, hyaline, smooth, with an entire sheath. Asexual morph where known sporodochial, monodictys-like hyphomycetes.

Type genus: Parabambusicola Kaz. Tanaka & K. Hiray.

Notes: This family superficially resembles Bambusicola, but can be distinguished from the latter by the ascomata surrounded by stromatic tissue (Parabambusicola) or compressed necks with wide ostioles (Aquastroma), and the multiseptate, clavate to fusiform, hyaline ascospores.

Aquastroma Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811325.

Etymology: From the Latin aqua meaning water and stroma.

Ascomata scattered to grouped, immersed to erumpent, globose in surface view, depressed globose in section. Ostiolar neck central, compressed, composed of thick-walled, heavily melanised, subglobose cells, with a wide ostiole. Ascomatal wall composed of pale brown, thin-walled cells. Pseudoparaphyses septate, branched and anastomosed. Asci fissitunicate, clavate, stipitate, with 8 ascospores. Ascospores clavate to fusiform, slightly curved, with a supramedian primary septum, multiseptate, slightly constricted at septa, hyaline, smooth, with an entire sheath. Asexual morph unknown.

Type species: Aquastroma magniostiolata Kaz. Tanaka & K. Hiray.

Notes: Aquastroma is superficially similar to Quintaria in having multiseptate ascospores and aquatic habitat. However, the type species of Quintaria, Q. lignatilis known from marine habitats, has long-necked ascomata with black incrustations surrounding the sides of the ostiolar canal (Zhang et al. 2012), and is phylogenetically close to members of the family Amniculicolaceae (Shearer et al. 2009) or the Testudinaceae/Platystomaceae (Suetrong et al., 2009, Zhang et al., 2012). Quintaria is regarded as a polyphyletic genus, because Q. submersa known from freshwater habitats does not cluster with Q. lignatilis and forms a sister clade to the Lindgomycetaceae (Zhang et al. 2012). An asexual morph was not observed for Aquastroma in culture, but this genus forms a clade with two hyphomycetes, Monodictys species, and their relationship is highly supported (100 %; Fig. 1). Monodictys, however, is not monophyletic and species in this genus are phylogenetically distributed in three ascomycete classes, Dothideomycetes, Leotiomycetes, and Sordariomycetes (see Notes in Monodictys). Aquastroma should be retained as a distinct genus, until sequence data of the type species of Monodictys (M. putredinis) become available to clarify the phylogenetic placement of Monodictys s. str.

Aquastroma magniostiolata Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811326. Fig. 40.

Fig. 40.

Aquastroma magniostiolata. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F, G. Asci; H–M. Ascospores (arrowheads indicate gelatinous sheath; J. in Black-Blue ink; K–M. in India ink); N. Germinating ascospore. A–E, G–J, N from KT 2485; F, K–M from culture KT 2485. Scale bars: A = 1 mm; B = 500 μm; C = 100 μm; D–N = 10 μm.

Etymology: From the Latin mangi-, meaning large, and ostiolatus, meaning ostiolate.

Ascomata scattered to 3–4 grouped, immersed to erumpent, globose in surface view, depressed globose in section, 180–250 μm high, 260–340 μm diam. Ostiolar neck central, compressed, 60–750 μm long, 75–130 μm wide, composed of thick-walled, heavily melanised, subglobose cells (2–3 μm diam), with a wide ostiole. Ascomatal wall in longitudinal section 25–30 μm thick at sides, composed of 5–7 layers of polygonal, thin-walled, 6–17 × 2.5–7.5 μm, pale brown cells; wall at the base 10–18 μm thick or poorly developed, composed of thin-walled flattened cells. Pseudoparaphyses septate, branched and anastomosed, 2–3 μm wide. Asci fissitunicate, clavate, 112.5–137.5 × 25–29.5 μm, with a stipe of 22–28 μm long, with 8 biseriate to triseriate ascospores. Ascospores clavate to fusiform, slightly curved, 30–44 × 9–13.5 μm (av. 36.5 × 10.3 μm, n = 39), l/w 2.8–4.5 (av. 3.6, n = 39), with a supramedian primary septum (0.40–0.47; av. 0.43, n = 39), 6–8-septate (2+1+3, 2+1+4, 3+1+3, 3+1+4), slightly constricted at septa, hyaline, smooth, with an entire sheath; sheath delimited and 2–3 μm wide at first, diffuse and up to 10 μm wide at a later.

Colonies on PDA (after 4 wk) attaining a diam of 3.2–3.4 cm, greenish grey; reverse almost black; no pigment produced. In culture sexual morph formed. Asci and ascospores larger than those on the natural host. Asci 130–170 × 22.5–37.5 μm (av. 148.9 × 29.9 μm, n = 30). Ascospores 39–47.5 × 11–16 μm (av. 43.5 × 13.5 μm, n = 50), l/w 2.8–3.8 (av. 3.2, n = 50).

Specimen examined: Japan, Mie, Tsu, Kamihama, Hyourtan pond, on submerged twigs of woody plant, 1 Jun. 2008, K. Tanaka & H. Yonezawa (holotype KT 2485 = HHUF 30122, culture ex-type CBS 139680 = JCM 19429 = MAFF 243824).

Notes: Aquastroma magniostiolata is shown to be morphologically distinct from Quintaria aquatica by its ascospores with 6–8 septa [vs. (10–)11–13(–14) septa in Q. aquatica; Hyde & Goh 1999]. It is also different from Q. microspora in its larger ascospores (vs. 26–31 × 5–5.5 μm in Q. microspora; Zhang et al. 2008a).

Parabambusicola Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811327.

Etymology: After its phylogenetic similarity to Bambusicola.

Ascomata mostly grouped, immersed to erumpent, subglobose in surface view, hemispherical with flattened base in section. Ostiolar neck short papillate, central. Ascomatal wall at the sides composed of pale brown, flattened cells; at the rim, composed of parallel rows of rectangular to polygonal cells. Pseudoparaphyses numerous. Asci broadly cylindrical to clavate, short-stalked, 8-spored. Ascospores fusiform, multiseptate, primary septum mostly supramedian, hyaline, smooth, with an entire sheath. Asexual morph unknown.

Type species: Parabambusicola bambusina (Teng) Kaz. Tanaka & K. Hiray.

Notes: A new genus Parabambusicola is established to accommodate P. bambusina formerly classified in Massarina (Teng, 1936, Aptroot, 1998, Tanaka and Harada, 2003b). Parabambusicola differs from Massarina in its hemispherical to depressed globose ascomata surrounded by stromatic tissue and without prominent clypeus, broadly cylindrical asci, and fusiform ascospores with several septa. It is distantly related to Massarina (Fig. 1).

Parabambusicola bambusina (Teng) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811392. Fig. 41.

Fig. 41.

Parabambusicola bambusina. A, B. Ascomata on the natural host surface; C. Ascomata in culture (on rice straw); D, E. Ascomata in longitudinal section; F. Ascomatal wall; G. Pseudoparaphyses; H. Ascus; I, J. Ascospores (arrowheads indicate gelatinous sheath); K. Spermatia. A, B from KH 139; C, G–J from culture KH 139; D–F from KT 2637; K from culture KT 2637. Scale bars: A = 1 mm; B, C = 500 μm; D, E = 100 μm; F–K = 10 μm.

Basionym: Massarina bambusina Teng, Sinensia 7: 512. 1936.

Specimens examined: Japan, Hokkaido, Chitose, Shikotsuko, on dead twigs of Sasa kurilensis, 5 Sep. 2001, Y. Harada, H 4321 = HHUF 26590, culture MAFF 239462; Iwate, Hachimantai, Aspite line, on dead twigs of Sasa kurilensis, 25 Jul. 2009, K. Tanaka & Y. Harada, KT 2637 = HHUF 30120, culture MAFF 243822; Aomori, Hirosaki, Tokiwano, Kuromori, Top of Mt. Iwaki, on dead twigs of Sasa sp., 21 Jun. 2008, K. Hirayama & K. Tanaka, KH 139 = HHUF 30121, culture MAFF 243823.

Note: For further information on this species, see Teng (1936) and Tanaka & Harada (2003b).

Periconiaceae (Sacc.) Nann., Repert. mic. uomo 4: 482. 1934. emend.

= Periconieae Sacc., Syll. Fung. 4: 235. 1886.

Ascomata scattered to grouped, immersed to erumpent, globose. Ostiolar neck central, papillate, with hyaline periphyses. Ascomatal wall in longitudinal section composed of several layers of thin-walled, pale brown to brown cells. Pseudoparaphyses cellular, branched, anastomosed. Asci fissitunicate, oblong to cylindrical, 8-spored. Ascospores broadly fusiform, 1-septate, hyaline, smooth, with an entire sheath. Conidiomata periconia- or noosia-like. Conidiophores macronematous, mononematous, sometimes lacking. Branches pale brown to brown, smooth to slightly echinulate. Conidial heads spherical. Conidiogenous cells monoblastic to polyblastic, discrete on stipe and branches. Conidia globose to ellipsoidal, aseptate, catenate, brown, verruculose to echinulate.

Type genus: Periconia Tode.

Note: Although Periconiaceae (Nannizzi 1934) has long been ignored in modern fungal systematics, it should be regarded as a natural taxon in the suborder Massarineae.

Periconia Tode, Fung. mecklenb. sel. (Lüneburg) 2: 2. 1791.

Type species: Periconia lichenoides Tode.

Notes: Among more than 185 taxa described as Periconia (http://www.indexfungorum.org, Aug. 2015), only two species have been reported to have sexual morphs. They are P. igniaria (the sexual morph was referred to as Didymosphaeria or Massarina in the Dothideomycetes; Booth, 1968, Aptroot, 1998) and P. prolifica (Remispora or Okeanomyces in the Sordariomycetes; Kohlmeyer, 1969, Pang et al., 2004), and therefore it is obvious that the genus Periconia is not monophyletic. DNA sequence data of the type species of this genus, P. lichenoides, are currently unavailable, but we judged that the lineage of Periconia in the Dothideomycetes, including typical members of Periconia, such as P. byssoides, P. cookei, P. igniaria, and P. digitata, corresponds to Periconia s. str., based on their morphological similarities with P. lichenoides (Mason & Ellis 1953). In contrast, P. prolifica, a marine fungus in the Sordariomycetes, does not have macronematous conidiophores (Anastasiou, 1963, Kohlmeyer, 1969), and is not regarded as Periconia s. str.

Periconia homothallica Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811328. Fig. 42.

Fig. 42.

Periconia homothallica. A, B. Ascomata on the natural host surface; C. Ascoma in culture (on rice straw); D. Ascoma in longitudinal section (in lactophenol cotton blue); E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath; J. in India ink); K. Germinating ascospores. A, B, D–H, K from KT 916; C, I, J from culture KT 916. Scale bars: A = 2 mm; B, C = 500 μm; D = 50 μm; E–K = 10 μm.

Etymology: Referring to its homothallism.

Ascomata scattered, immersed to erumpent, globose, 140–190 μm high, 160–180 μm diam, with an ostiole. Ostiolar neck central, papillate, 24–40 μm long, 37–62 μm wide, with hyaline periphyses. Ascomatal wall in longitudinal section uniformly 11–15 μm thick, composed of 4–6 layers of polygonal, thin-walled, 3–15 × 2–5 μm, pale brown cells. Pseudoparaphyses cellular, 2–2.5 μm wide at the apex, up to 5 μm wide at the base, branched, anastomosed. Asci fissitunicate, cylindrical to lageniform, 85–119.5 × 13–17.5 μm (av. 96.5 × 15.3 μm, n = 20), with a shallow ocular chamber, short-stalked (3.5–6 μm long), with 8 biseriate ascospores. Ascospores broadly fusiform, 22–31 × 7–10 μm (av. 26.3 × 8.7 μm, n = 60), l/w 2.6–3.7 (av. 3.0, n = 60), with a nearly median septum (0.48–0.53; av. 0.51, n = 38), hyaline, smooth, with an entire sheath; sheath gelatinous, up to 10 μm wide when fresh, later 1–2 μm wide.

Colonies on PDA (after 2 wk) attaining a diam of 4.3–5.6 cm, straw; reverse similar; no pigment produced. In culture sexual morph formed. Ascospores slightly smaller than those on the natural host, 20–28 × 6–9 μm (av. 24.0 × 7.8 μm, n = 45), l/w 2.7–3.5 (av. 3.1, n = 45).

Specimen examined: Japan, Yamagata, Mogamigun, Sakekawa, Magarikawa-river, riverbank, on dead leaves of Phragmites japonica, 13 Oct. 2002, Y. Ooki & Y. Harada (holotype KT 916 = HHUF 29105, culture ex-type CBS 139698 = JCM 13100 = MAFF 239610).

Notes: Although we were not able to observe a typical periconia-like asexual morph from our specimen and isolate, we introduce this as a new species of Periconia based on the morphology of the sexual morph. The above material shares many characteristics with sexual species in Periconia s. str. (i.e., P. igniaria and P. pseudodigitata), such as those with immersed globose ascomata with papillate necks, a peridium composed of thin-walled polygonal cells, cylindrical asci, and broadly fusiform hyaline ascospores with nearly median septum. Periconia homothallica, however, has shorter but wider asci than those of P. igniaria (85–119.5 × 13–17.5 μm vs. 150–165 × 8–12 μm, Booth 1968) and has wider ascospores than those of P. pseudodigitata [22–31 × 7–10 μm vs. 19.5–27(–32) × 5–7 μm].

In our phylogenetic tree (Fig. 1), P. homothallica formed an unsupported clade with P. igniaria, as well as species in other hyphomycetous genera, such as “Sporidesmium” tengii that have macronematous conidiophores and multiseptate conidia (Wu & Zhuang 2005), and Noosia banksiae, a species that lacks conspicuous conidiophores (Crous et al. 2011a). The presence of these genera in this lineage may indicate that Periconia should be subdivided into several morphologically similar genera or that the taxonomic status of these Noosia/Sporidesmium isolates should be re-evaluated.

Periconia pseudodigitata Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811329. Fig. 43.

Fig. 43.

Periconia pseudodigitata. A, B. Ascomata on the natural host surface; C. Ascomata in longitudinal section (in lactophenol cotton blue); D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores (arrowheads indicate gelatinous sheath; H. in Black-Blue ink); J. Germinating ascospore; K. Conidiomata in culture (on rice straw); L–O. Conidial heads and conidiophores; P, Q. Conidia. A, B, E, G–J from KT 1395; C, D, F from KT 644; K–M from culture KT 1195; N from culture KT 644; O–Q from culture KT 1395. Scale bars: A = 1 mm; B, K = 500 μm; C = 100 μm; D–J, P, Q = 10 μm; L–O = 20 μm.

Etymology: After its morphological and phylogenetic similarity to Periconia digitata.

Ascomata numerous, scattered or 2–3 grouped, immersed to erumpent, globose, 160–200 μm high, 130–250 μm diam. Ostiolar neck central, papillate, 45–55 μm long, 45–65 μm wide, with hyaline periphyses, surrounded by clypeus-like structure composed of subglobose to polygonal, slightly thickened, 5–7 × 3–4 μm, dark brown cells. Ascomatal wall in longitudinal section 8–13 μm thick at side, 5–8 μm thick at the base, composed of 3–5 layers of thin-walled, 6–13 × 2–5 μm, brown cells. Pseudoparaphyses cellular, numerous, 2–2.5 μm wide at the apex, 4–6 μm wide at the base, septate, branched, anastomosed, guttulate, associated with gelatinous material. Asci fissitunicate, cylindrical, 70–110 × 10.5–15.5 μm (av. 88.4 × 12.2 μm, n = 33), rounded at the apex and with an apical chamber, short-stalked (5–15 μm long), with 8 irregularly biseriate ascospores. Ascospores broadly fusiform with rounded ends, straight or slightly curved, 19.5–27(–32) × 5–7 μm (av. 22.5 × 6.1 μm, n = 134), l/w 2.9–4.5 (av. 3.7, n = 134), with almost median septum (0.48–0.55, av. 5.1, n = 36), slightly constricted at the septum, hyaline, with or without guttules, smooth, with an entire sheath; sheath gelatinous, 1–2 μm wide at side and 2–4 μm wide at both ends in fresh, becoming delimited sheath in dry condition. Senescent spores brown, echinulate, 1-septate. Ascospores germinating from one or both ends.

Colonies on PDA (after 4 wk) attaining a diam of 3.2–4.6 cm, straw to amber with white margin; reverse pale luteous; no pigment produced. In culture conidial and spermatial morphs formed. Conidiophores macronematous, mononematous, single or rarely 2–3 together on stromata, 250–450 μm long, mostly 8.5–15 μm wide (16–24 μm wide at basal stromatic portion), 4–7-septate at 35–90 μm long intervals, with several branches. Branches 10–50 μm long, septate, pale brown to brown, slightly echinulate. Conidial heads spherical, ca. 60–120(–150) μm diam. Conidiogenous cells monoblastic, discrete on stipe and branches. Conidia catenate, globose, brown, verruculose to shortly echinulate, 7–9.5 μm (av. 8.3 μm, n = 90). Spermogonia globose to subglobose, single to gregarious, uniloculate to multiloculate, 140–340 μm diam. Spermatia subglobose, hyaline, smooth, 3–5 × 1.5–2 μm (av. 3.7 × 1.7 μm, n = 30).

Specimens examined: Japan, Aomori, Hirakawa, Hirakawa-river, riverbank, on dead culms of Phragmites australis, 5 Aug. 2001, K. Tanaka, KT 644 = HHUF 27569, culture JCM 13164 = MAFF 239674; Aomori, Hirosaki, Kawai, Hirakawa-river, riverbank, 18 Aug. 2001, K. Tanaka, KT 680 = HHUF 27570; Aomori, Hirosaki, Shimizumori, Oowasawa-river, riverbank, 14 Aug 2001, K. Tanaka, KT 673 = HHUF 27571; Hokkaido, Akkeshi, Toraibetsu-river, on submerged stems of herbaceous plant, 3 Jun. 2003, K. Tanaka & S. Hatakeyama, KT 1195A = HHUF 29368, culture JCM 13165 = MAFF 239675; ibid., on dead culms of Phragmites australis, 7 Sep. 2003, K. Tanaka & S. Hatakeyama (holotype KT 1395 = HHUF 29370, culture ex-type CBS 139699 = JCM 13166 = MAFF 239676); ibid., KT 1396–1398 = HHUF 29371–29373.

Notes: Periconia pseudodigitata is phylogenetically and morphologically closest to P. digitata, but the conidial dimensions of P. pseudodigitata (7–9.5 μm) are slightly smaller than those reported for P. digitata by Ellis (1971; 7–11 μm). Sequence similarity between these two taxa in the ITS region was 95.8 % (503/525) with 0.8 % (4/525) gaps. Periconia pseudodigitata superficially resembles P. igniaria (= Massarina igniaria), which also has an ascomatal morph. The sexual morph of P. pseudodigitata, however, differs from that of P. igniaria (Booth 1968) by its narrower ascospores (5–7 μm vs. 8–9 μm) surrounded by a mucilaginous sheath, and shorter but wider asci (70–110 × 10.5–15.5 μm vs. 150–165 × 8–12 μm) with biseriate ascospores. The asexual morph of P. pseudodigitata has verruculose to shortly echinulate conidia, while that of P. igniaria has conidia with conspicuous spines about 1 μm long (Mason and Ellis, 1953, Ellis, 1971, Matsushima, 1971, Bunning and Griffiths, 1984).

Sulcatisporaceae Kaz. Tanaka & K. Hiray., fam. nov. MycoBank MB814431.

Etymology: Referring to the name of the type genus.

Ascomata scattered to grouped, immersed to erumpent, globose in surface view, subglobose to hemispherical in section. Ostiolar neck short papillate, central, with periphyses. Ascomatal wall composed of several layers of compressed cells, poorly developed at the base. Pseudoparaphyses trabeculate, branched and anastomosed. Asci clavate, short-stalked, 8-spored. Ascospores broadly fusiform, 1-septate, hyaline, with an entire sheath. Conidiomata pycnidial, globose. Conidiogenous cells cylindrical to doliiform, annellidic or phialidic. Conidia ellipsoid to subglobose, hyaline to dark brown, 1- to multi-septate (occasionally muriform), with or without striate ornamentation.

Type genus: Sulcatispora Kaz. Tanaka & K. Hiray.

Note: A new family, Sulcatisporaceae is established here for Magnicamarosporium and Sulcatispora genera nova and Neobambusicola (Crous et al. 2014b).

Magnicamarosporium Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811292.

Etymology: After its morphological similarity to Camarosporium and its large-sized conidiomata.

Conidiomata pycnidial, scattered, immersed, depressed globose in section. Ostiolar neck central, papillate to short cylindrical. Paraphyses filamentous, sometimes branched, associated with gelatinous material. Conidiophores absent. Conidiogenous cells holoblastic, cylindrical to doliiform. Conidia subglobose to obovoid, sometimes obpyriform, muriform, dark brown. Sexual morph unknown.

Type species: Magnicamarosporium iriomotense Kaz. Tanaka & K. Hiray.

Notes: Several coelomycetous genera are known to have dictyosporous pigmented conidia and their taxonomic revision has been reported (e.g., Van Warmelo and Sutton, 1981, Nag Raj, 1989, Verkley et al., 2005, Crous et al., 2015b). Amarenographium is characterised by longer and branched conidiophores and conidia with cap-like gelatinous appendages (Eriksson 1982). Species in this genus are reported from marine environments (Kohlmeyer & Volkmann-Kohlmeyer 1991) or palms (Taylor & Hyde 2003), and may be genetically close to the Trematosphaeriaceae (Hodhod et al. 2012). Camarosporellum and Camarographium have distoseptate conidia (Sutton, 1980, Verkley et al., 2005) rather than euseptate conidia like Magnicamarosporium. Camarographium is heterogenous, and C. koreanum belongs to Unknown Clade III of the Massarineae (Fig. 1), while C. carpini may have affinity with the Sporormiaceae based on a BLAST search using LSU and ITS nrDNA sequences (Crous et al. 2011b). Camarosporium, to some degree resembles our new genus, but Magnicamarosporium has larger conidiomata with a conspicuous projecting ostiole composed of thick-walled cells, long paraphyses with a gelatinous coating, and larger dark brown conidia than Camarosporium. In addition, Camarosporium does not have phylogenetic affinity with Magnicamarosporium, because the type species of Camarosporium (C. quaternatum) is a member of the Pleosporineae, based on molecular data (Crous et al., 2006, Wijayawardene et al., 2014a, Wijayawardene et al., 2014b, Wijayawardene et al., 2014d, Crous et al., 2015b).

The phylogenetic tree generated in this study indicated that Magnicamarosporium and Neobambusicola cluster together with 74 % BP (Fig. 1). However, the conidial morphology of Neobambusicola (fusoid-ellipsoid and 1-septate; Crous et al. 2014b) is quite distinct from that of Magnicamarosporium.

Magnicamarosporium iriomotense Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811293. Fig. 44.

Fig. 44.

Magnicamarosporium iriomotense. A. Immersed conidiomata and discharged conidia lying on the natural host surface; B. Conidiomata produced in culture (on rice straw); C, D. Conidiomata in longitudinal section (C. on nature; D. in culture); E. Conidiomatal wall; F. Paraphyses; G. Conidiogenous cell (in trypan blue); H–L. Conidia; M. Immature conidium; N. Germinating conidium. A, C, E, H–J, N from KT 2822; B, D, F, G, K–M from culture KT 2822. Scale bars: A, B = 500 μm; C, D = 50 μm; E–N = 10 μm.

Etymology: Referring to the location where the specimen was collected.

Conidiomata pycnidial, scattered, deeply immersed, depressed globose in section, 330–440 μm high, 700–760 μm diam. Ostiolar neck central, papillate to short cylindrical, 120–150 μm long, 80–100 μm wide, composed of polygonal to rectangular, thick-walled, dark brown cells of 2–5 μm diam. Conidiomatal wall in longitudinal section uniformly 10–20 μm thick, composed of 3–6 layers of polygonal to subglobose, thin-walled, pale brown cells of 2–7.5 × 2–5 μm. Paraphyses sometimes branched, 20–50(–80) μm long, 1.5–2.5 μm wide, associated with gelatinous material. Conidiophores absent. Conidiogenous cells holoblastic, 7–11 × 4.5–6 μm, cylindrical to doliiform. Conidia subglobose to obovoid, sometimes obpyriform, 29–40.5(–42.5) × 19.5–25(–27) μm (av. 34.7 × 22.2 μm, n = 100), l/w 1.2–2.0 (av. 1.6, n = 100), with 4–6 trans- and 1–2 vertical-septa, frequently with oblique septa, dark brown, smooth, without sheath.

Colonies on PDA (after 4 wk) attaining a diam of 1.1–1.5 cm, olivaceous black with buff margin; reverse buff to honey; no pigment produced. In culture asexual morph formed. Conidiomata pycnidial, 130–170 μm diam, scattered to 2–3 grouped, globose to subglobose. Conidia 29.5–43 × 18–23 μm (av. 35.3 × 20.3 μm, n = 31), l/w 1.5–2.1 (av. 1.7, n = 31). Sexual morph unknown.

Specimen examined: Japan, Okinawa, Isl. Iriomote, Tropical botanic garden, on dead twigs of Diplospora dubia, 13 Jul. 2011, K. Tanaka & K. Hirayama (holotype KT 2822 = HHUF 30125, culture ex-type CBS 139696 = JCM 19402 = MAFF 243827).

Notes: There are more than 500 names in Camarosporium (http://www.indexfungorum.org, Aug. 2015). Many of these taxa have been described on the basis of their host associations (Sutton, 1980, Marincowitz et al., 2008), but there is no record of Camarosporium from host plants within the Rubiales, which includes Diplospora dubia, the host of M. iriomotense.

Sulcatispora Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811294.

Etymology: From the Latin sulcatus meaning “furrowed” and spora meaning “spore”, referring to the striate ornamentation of the conidia.

Ascomata scattered to grouped, immersed to erumpent, globose in surface view, subglobose to hemispherical in section. Ostiolar neck short papillate, central, with periphyses. Ascomatal wall composed of several layers of compressed cells, poorly developed at the base. Pseudoparaphyses trabeculate, branched and anastomosed. Asci fissitunicate, clavate, short-stalked. Ascospores broadly fusiform, 1-septate, hyaline, with an entire sheath. Conidiomata pycnidial, globose. Conidiogenous cells cylindrical, annellidic. Conidia ellipsoid, yellowish brown, multiseptate, with striate ornamentation.

Type species: Sulcatispora acerina Kaz. Tanaka & K. Hiray.

Notes: Ascomatal features of Sulcatispora are most similar to those of Massarina s. str. (Massarinaceae) but the ascomata lack a prominent clypeus, in contrast to species within Massarina s. str. The most distinctive feature of Sulcatispora is the striated conidia. Some species in Phaeophleospora (e.g., P. striae; Taylor & Hyde 2003) are known to have such conidia, but Phaeophleospora (type species: P. eugeniae) is phylogenetically placed in the Mycosphaerellaceae of the Capnodiales (Crous et al., 2007, Crous et al., 2009b). Sclerostagonospora also has species with conidia with striate ornamentation (e.g., S. opuntiae; Huhndorf 1992), but species of Sclerostagonospora have phylogenetic affinities with members of the Phaeosphaeriaceae, such as “Phoma” caloplacae and “Phoma” foliaceiphila (Crous et al., 2011a, Lawrey et al., 2012), and may have phaeosphaeria-like sexual morphs (Quaedvlieg et al. 2013). Sulcatispora is phylogenetically related to Bambusicola, which also has a similar asexual morph. Bambusicola, however, has conical ascomata with flattened bases, narrower asci (up to 14 μm wide), and narrowly fusiform ascospores (Dai et al. 2012).

Sulcatispora acerina Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811295. Fig. 45.

Fig. 45.

Sulcatispora acerina. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–K. Ascospores (arrowheads indicate mucilaginous sheath); L. Germinating ascospore; M, N. Conidiomata in culture (on rice straw); O. Conidioma in longitudinal section; P. Conidiomatal wall; Q. Conidiogenous cells (arrowheads indicate annellations); R–W. Conidia. A–L from KT 2982; M–W from culture KT 2982. Scale bars: A = 1 mm; B, M, N = 500 μm; C, O = 50 μm; D–L, P–W = 10 μm.

Etymology: Referring to the generic name of the host.

Ascomata scattered, immersed to erumpent, globose in surface view, depressed globose to hemispherical in section, 200–250 μm high, 260–450 μm diam. Ostiolar neck inconspicuous short papillate, 60–85 μm diam, central, with periphyses. Ascomatal wall in longitudinal section 12–25 μm thick at sides, composed of polygonal, 5–8 × 2–3 μm, compressed cells, surrounded by short, brown hyphae (2.5–3.5 μm thick), poorly developed at the base. Pseudoparaphyses trabeculate, 1.5–2 μm wide, branched and anastomosed, associated with gelatinous material. Asci fissitunicate, clavate, 87–113 × 15–20 μm (av. 98.4 × 17.1 μm, n = 17), rounded at the apex, with an apical chamber and faint ring, short-stalked (15–22 μm long). Ascospores broadly fusiform, slightly acute at the apex, (23–)26–32 × 7–9 μm (av. 28.5 × 7.7 μm, n = 50), l/w 3.2–4.2 (av. 3.7, n = 50), with a submedian primary septum (0.50–0.57; av. 0.53, n = 50), hyaline, surrounded by an entire sheath; sheath 2–3 μm wide at first, later diffuse, 4–5 μm wide.

Colonies on PDA (after 4 wk) attaining a diam of 2.1–2.4 cm, grey olivaceous to honey; reverse olivaceous to black; no pigment produced. In culture asexual morph formed. Conidiomata pycnidial, 180–270 μm high, 160–240 μm diam, immersed, scattered, globose, ostiolate. Ostiolar neck short papillate, 25–45 μm long, 35–45 μm diam. Conidiomatal wall uniformly 10–20 μm thick, composed of 4–7 layers of polygonal, 6–10 × 2–3 μm, dark brown cells. Conidiophores absent. Conidiogenous cells cylindrical to lageniform, 10–20 μm long, 2 μm wide at the apex, 3.5–5.5 μm wide at the base, annellidic. Conidia ellipsoid, rounded at the apex, truncate at the base, yellowish brown, 20–28(–30) × 6.5–8 μm (av. 25.9 × 7.2 μm, n = 50), l/w 3.1–4.2 (av. 3.6, n = 50), 3(–5)-septate (1+1+1, rarely 2+1+2), with striate ornamentation.

Specimen examined: Japan, Fukuoka, Kasuya, Hisayama, Yamada, Mt. Tachibanayama, on dead twigs of Acer palmatum, 31 Mar. 2012, K. Tanaka (holotype KT 2982 = HHUF 30449, culture ex-type CBS 139703).

Note: The conidia of S. acerina are very similar to those of S. berchemiae, but S. acerina can be distinguished from the latter by its larger ascospores [(23–)26–32 × 7–9 μm vs. 22–29 × 5.5–7 μm].

Sulcatispora berchemiae Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811296. Fig. 46.

Fig. 46.

Sulcatispora berchemiae. A, B. Ascomata on the natural host surface; C, D. Ascomata in longitudinal section; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate mucilaginous sheath); K. Germinating ascospore; L. Conidiomata in culture (on rice straw); M. Conidioma in longitudinal section; N. Conidiomatal wall; O. Conidiogenous cells (arrowheads indicate annellations); P–S. Conidia. A–K from KT 1607; L–S from culture KT 1607. Scale bars: A = 1 mm; B, L = 500 μm; C, D = 100 μm; E–K, M–S = 10 μm.

Etymology: Referring to the generic name of the host.

Ascomata scattered to 2–4 grouped, immersed to erumpent, globose in surface view, subglobose to hemispherical in section, 150–280 μm high, 350–530 μm diam. Ostiolar neck short papillate, ca. 60 μm diam, central, with periphyses. Ascomatal wall in longitudinal section 10–18 μm thick at sides, composed of 3–6 layers of 7–18 × 2 μm compressed cells, poorly developed at the base. Pseudoparaphyses trabeculate, 1.5–2.5 μm wide, branched and anastomosed, associated with gelatinous material. Asci fissitunicate, clavate, 77.5–100 × 13.5–18 μm (av. 90.1 × 15.2 μm, n = 20), rounded at the apex, with an apical chamber and faint ring, short-stalked (13–15 μm long). Ascospores broadly fusiform, 22–29 × 5.5–7 μm (av. 26.0 × 6.4 μm, n = 50), l/w 3.5–4.4(–4.7) (av. 4.1, n = 50), with a submedian primary septum (0.50–0.57; av. 0.53, n = 49), hyaline, surrounded by an entire sheath; sheath delimited, 2 μm wide at first, later diffuse, up to 6 μm wide.

Colonies on PDA (after 4 wk) attaining a diam of 2.7–2.8 cm, white to buff; reverse buff to greyish sepia; no pigment produced. In culture asexual morph formed. Conidiomata pycnidial, 90–130 μm high, 90–150 μm diam, immersed, scattered, globose to subglobose, ostiolate. Ostiolar neck short papillate. Conidiomatal wall uniformly 12–17.5 μm thick, composed of flattened, thin-walled, polygonal cells (3.5–6 × 1.5–2.5 μm). Conidiophores absent. Conidiogenous cells cylindrical, 12–15 μm long, 2 μm wide at the apex, 4–8 μm wide at the base, annellidic. Conidia ellipsoid, 20–30 × 6.5–8 μm (av. 24.8 × 7.1 μm, n = 50), l/w 2.9–4.2 (av. 3.5, n = 50), rounded at the apex, truncate at the base, yellowish brown, 3(–5)-septate (1+1+1, rarely 2+1+2), with striate ornamentation.

Specimens examined: Japan, Aomori, Towada, Sanbongi, Yagami, on vines of Berchemia racemosa, 2 Dec. 2003, K. Tanaka, S. Hatakeyama & N. Nakagawara (holotype KT 1607 = HHUF 29097, culture ex-type CBS 139704 = JCM 13101 = MAFF 239611); ibid., KT 1608 = HHUF 29098.

Note: This species is closely related to S. acerina with regard to its LSU sequences (similarity 1295/1302 = 99.5 %), but the ITS sequence shows a great deal of variation compared to the ITS of S. acerina (similarity 770/850 = 90.6 %, with gaps 29/850 = 3.4 %).

Trematosphaeriaceae K.D. Hyde et al., Cryptog. Mycol. 32: 347. 2011.

Type genus: Trematosphaeria Fuckel.

Trematosphaeria Fuckel, Jb. nassau. Ver. Naturk. 23–24: 161. 1870.

Type species: Trematosphaeria pertusa Fuckel.

Notes: Although more than 200 species have been assigned to Trematosphaeria (http://www.indexfungorum.org, Aug. 2015), most of these species have not had their generic placements verified by molecular evidence. The type species of this genus, T. pertusa, usually grows on terrestrial wood, but also can survive within freshwater (Suetrong et al. 2011b). Recently, Madurella grisea, originating from a human infection, water and pastry gel, was added to Trematosphaeria based on a multi-gene phylogeny (Ahmed et al. 2014).

Trematosphaeria pertusa Fuckel, Jb. Nassau. Ver. Naturk. 23–24: 161. 1870. Fig. 47.

Fig. 47.

Trematosphaeria pertusa. A, B. Ascomata on the natural host surface; C. Ascoma in longitudinal section; D. Ascomatal wall; E. Pseudoparaphyses; F. Ascus; G–I. Ascospores; J. Germinating ascospore; K. Spermogonia in culture; L. Spermatiophores; M. Spermatia. A–J from KT 1496; K–M from culture KT 1496. Scale bars: A, K = 500 μm; B = 200 μm; C = 50 μm; D–J, L, M = 10 μm.

Ascomata gregarious, immersed or becoming superficial by weathering of host tissue, globose to pyriform in section, 270–380 μm high, 190–350 μm diam. Ostiolar neck central, papillate to cylindrical, 70–140 μm long, 90–130 μm wide, composed of thick-walled, heavily melanised cells (2–4 μm diam), with periphyses. Ascomatal wall in longitudinal section uniformly 17–25 μm thick, composed of 6–9 layers of polygonal to rectangular, thin-walled, brown cells (2–15 × 2–5 μm). Pseudoparaphyses branched and anastomosed, 1.5–2.5 μm wide. Asci fissitunicate, clavate to cylindrical, 73.5–102.5 × 12.5–17 μm (av. 89.8 × 14.6 μm, n = 20), with a stipe of 8.5–22 μm long, with 8 biseriate ascospores. Ascospores fusiform, slightly curved, 20–27.5 × 6–8 μm (av. 23.9 × 7.1 μm, n = 50), l/w 2.8–4.0 (av. 3.4, n = 50), with a nearly median or somewhat supramedian primary septum (0.43–0.53; av. 0.48, n = 50), rarely 3-septate, slightly constricted at the septum, reddish brown, smooth, without sheath.

In culture spermatial morph formed. Spermatia subglobose, hyaline, 2.8–3.9 × 1.9–2.3 μm (av. 3.4 × 2.1 μm, n = 20), l/w 1.4–1.9 (av. 1.6, n = 20).

Specimens examined: Japan, Aomori, Hirosaki, Aoki, Mohei-pond, on submerged twigs of woody plant, 27 Sep. 2003, K. Tanaka & N. Asama, KT 1496 = HHUF 30153, culture JCM 19430 = MAFF 243879; Hokkaido, Isl. Rebun, Nairo, Nairo-river, on submerged twigs of woody plant, 16 Aug. 2013, K. Tanaka, KT 3314 = HHUF 30452, culture CBS 139705; ibid., KT 3315 = HHUF 30453, culture CBS 139706.

Notes: We identified our specimens/isolates as T. pertusa, the type species of Trematosphaeria, based on close similarities between sequences obtained from the above isolates and an ex-epitype strain of the species (CBS 122368; Ahmed et al. 2014), i.e., 484/485 (99.8 %) in ITS (GenBank KF015668) and 900/907 (99.2 %) in tef1 (GenBank KF015701) regions. In comparison with the description of T. pertusa based on the neotype (Zhang et al. 2008b), our specimens have shorter asci (73.5–102.5 μm vs. 100–145 μm) and ascospores (av. 20–27.5 μm vs. 27.5–32.5 μm). Further collections of this species are needed to clarify the taxonomic significance of the intraspecific morphological variation.

Unknown Clade I

Fuscostagonospora Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811330.

Etymology: From the Latin fusco-, meaning dark brown and the generic name, Stagonospora.

Ascomata scattered, immersed, globose to subglobose. Ostiolar neck clypeate, central, short papillate, with periphyses. Ascomatal wall composed of pale brown, compressed cells. Pseudoparaphyses appearing trabecular, branched and anastomosed, associated with gelatinous material. Asci fissitunicate, cylindrical, with a long stipe. Ascospores narrowly fusiform, 1(–3)-septate, hyaline, with an entire sheath. Conidiomata pycnidial, scattered, immersed, depressed globose, ostiolate. Conidiomatal wall composed of thin-walled cells. Conidiophores absent. Conidiogenous cells doliiform, annellidic. Conidia yellow to pale brown and 3-septate.

Type species: Fuscostagonospora sasae Kaz. Tanaka & K. Hiray.

Notes: The new genus, Fuscostagonospora, is introduced to accommodate the bambusicolous fungus, F. sasae. This genus is reminiscent of Stilbospora in having pigmented phragmosporous conidia, but the latter genus has acervular conidiomata filled with paraphyses and phylogenetically groups within the Diaporthales in the Sordariomycetes (Crous et al., 2012, Voglmayr and Jaklitsch, 2014). The conidial morphology of Fuscostagonospora is also similar to that of Sclerostagonospora, but the latter genus has phylogenetic affinity with species in the Phaeosphaeriaceae (Pleosporales) and may have a phaeosphaeria-like sexual morph (Quaedvlieg et al. 2013). In overall morphology, such as the narrowly fusiform ascospores and pigmented septate conidia, as well as in host preferences, Fuscostagonospora is most similar to Bambusicola (Bambusicolaceae; Dai et al. 2012). Fuscostagonospora, however, has ascomata with a prominent clypeus and phylogenetically deviates from the Bambusicolaceae, forming an Unknown Clade I (Fig. 1).

Fuscostagonospora sasae Kaz. Tanaka & K. Hiray., sp. nov. MycoBank MB811331. Fig. 48.

Fig. 48.

Fuscostagonospora sasae. A, B. Ascomata on the natural host surface; C, D. Ascomata in longitudinal section; E. Ascomatal wall; F. Pseudoparaphyses; G. Ascus; H–J. Ascospores (arrowheads indicate gelatinous sheath; H, I. in India ink); K. Germinating ascospore; L, M. Conidiomata in culture (on rice straw); N. Conidioma in longitudinal section; O. Conidiomatal wall; P, Q. Conidiogenous cells (arrowheads indicate annellations); R. Conidia. A–K, N, O from KT 1467; L, M, P–R from culture KT 1467. Scale bars: A, L = 1 mm; B, M = 500 μm; C, D, N = 100 μm; E–K, O–R = 10 μm.

Etymology: Referring to the host genus.

Ascomata scattered, immersed, globose to subglobose, 220–250 μm high, 400–450 μm diam. Ostiolar neck clypeate, central, short papillate, 50–65 μm long, 75–90 μm wide, with periphyses. Ascomatal wall in longitudinal section 10–15 μm thick at sides, composed of 3–4 layers of compressed, 5–10 × 2–3 μm, pale brown cells. Pseudoparaphyses appearing trabecular, 1–1.5 μm wide, branched and anastomosed, associated with gelatinous material. Asci fissitunicate, cylindrical, 90–140 × 8.5–14.5 μm (av. 107.6 × 9.8 μm, n = 30), with an apical chamber, with a relatively long stipe (20–32.5 μm long). Ascospores narrowly fusiform, 1(–3)-septate, 22–31.5(–36) × 3–5.5 μm (av. 27.5 × 4.1 μm, n = 25), l/w 5.6–8.3 (av. 6.8, n = 25), with a nearly median septum (0.48–0.57; av. 0.52, n = 20), hyaline, surrounded by an entire gelatinous sheath up to 7 μm wide.

Conidiomata on the natural host pycnidial, scattered, immersed, depressed globose in section, 250 μm high, 500 μm diam, ostiolate. Conidiomatal wall in longitudinal section 12–20 μm thick, composed of thin-walled, hyaline to pale brown cells. In culture same coelomycetous asexual morph formed. Conidiophores absent. Conidiogenous cells doliiform, annellidic, 5–12 × 3–7 μm (av. 9.7 × 5.3 μm, n = 20). Conidia hyaline and aseptate at first, later yellow to pale brown and 3-septate, 17–22.5 × 4–5 μm (av. 19.8 × 4.4 μm, n = 50), l/w (3.4–)3.7–5.5 (av. 4.5, n = 50).

Colonies on PDA (after 4 wk) attaining a diam of 3.7–4.7 cm, iron grey; reverse similar; no pigment produced. In culture asexual morph formed.

Specimen examined: Japan, Fukushima. Minamiaizu, Ose pond, on dead twigs of Sasa sp., 30 Aug. 2003, N. Asama (holotype KT 1467 = HHUF 29106, culture ex-type CBS 139687 = JCM 13104 = MAFF 239614).

Unknown Clade IV

Pseudoxylomyces Kaz. Tanaka & K. Hiray., gen. nov. MycoBank MB811332.

Etymology: After its morphological similarity to Xylomyces.

Colonies on natural substratum scattered, dark brown, glistening. Mycelium immersed in agar medium, pale brown to reddish brown. Stromata lacking. Conidiophores branched, septate, brown. Conidiogenesis holoblastic. Conidia broadly fusiform, with several trans-septa of thick-walled, yellowish brown to dark brown with paler end cells. Chlamydospores absent. Sexual morph unknown.

Type species: Pseudoxylomyces elegans (Goh et al.) Kaz. Tanaka & K. Hiray.

Notes: A new genus Pseudoxylomyces is proposed to accommodate Xylomyces elegans. The type species of Xylomyces (X. chlamydosporus) is known to have phylogenetic affinities with the freshwater genus, Jahnula in the Jahnulales (Campbell et al., 2007, Sivichai et al., 2011). Therefore, Xylomyces is now regarded as a synonym of Jahnula (Hyde et al. 2013). Xylomyces elegans was excluded from the genus (Suetrong et al. 2011a), because it did not cluster with the type species of Xylomyces in phylogenetic analyses using ITS (Prihatini et al. 2008) and SSU sequences (Shearer et al. 2009). However, no taxonomic decision has been made for X. elegans.

Xylomyces is characterised by its lack of conidiophores and conidiogenous cells but it produces large, dark, thick-walled, multiseptate, intercalary, narrowly fusiform chlamydospores (Goos et al., 1977, Goh et al., 1997). In contrast, Pseudoxylomyces typified by P. elegans is quite distinctive in producing broadly fusiform conidia holoblastically at the tip of the conidiophores (Fig. 49A, B).

Fig. 49.

Pseudoxylomyces elegans. A, B. Conidiophores and conidia; C–E. Conidia (E. bleached conidium); F. Germinating conidium. A, B, D, E from culture KT 2887; C, F from KT 2887. Scale bars: A = 200 μm; B–F = 20 μm.

Pseudoxylomyces elegans (Goh et al.) Kaz. Tanaka & K. Hiray., comb. nov. MycoBank MB811333. Fig. 49.

Basionym: Xylomyces elegans Goh et al., Mycol. Res. 101: 1324. 1997.

Mycelium immersed in agar, pale brown to reddish brown. Conidiophores up to 40 μm long. Conidiogenesis holoblastic. Conidia broadly fusiform, 4–7-septate, 72.5–98 × 35–41 μm, l/w 2.0–2.8, brown, with paler end cells. Sexual morph unknown.

Specimen examined: Japan, Okinawa, Isl. Iriomote, Oomijya-river, on submerged twigs of woody plant, 12 Jul. 2011, K. Tanaka & K. Hirayama, KT 2887 = HHUF 30139, culture MAFF 243852.

Notes: This species appears to be widely distributed and has been reported from Australia, Seychelles (Goh et al. 1997), Thailand (Sivichai et al. 2000), Hong Kong (Tsui & Hyde 2004), USA (Raja et al. 2007), Brazil (Barbosa & Gusmão 2011), and India (Patil & Borse 2015). A BLAST search using ITS sequences from our culture showed 98.7 % (464/470) similarity to sequences of X. elegans (GenBank FJ887920) collected from Thailand (Prihatini et al. 2008).

Unknown Clade VI

Monodictys S. Hughes, Canad. J. Bot. 36: 785. 1958.

Type species: Monodictys putredinis (Wallr.) S. Hughes, Canad. J. Bot. 36: 785. 1958.

Notes: More than 60 species have been described in Monodictys (http://www.indexfungorum.org, Aug. 2015), but their phylogenetic placements are mostly unknown. Apparently, Monodictys is a heterogenous group of hyphomycetes. Several species within this genus such as M. arctica (Leptosphaeriaceae; Day et al. 2006), M. cf. putredinis (Melanommataceae; Samuels 1980), Monodictys spp. (Parabambusicolaceae; Fig. 1), and M. capensis (Unknown Clade IV; Fig. 1) are scattered within the Dothideomycetes. Monodictys pelagica is known to have phylogenetic relationships with the Sordariomycetes (Mouzouras and Jones, 1985, Campbell et al., 2002), and Monodictys sp. is reported as an asexual morph of Hyaloscypha albohyalina var. monodictys (Hosoya & Huhtinen 2002; currently Hyaloscypha monodictys, Han et al. 2014) in the Leotiomycetes. Phylogenetic reassessment of many species of Monodictys including the type species of this genus (M. putredinis) is needed for taxonomic revision of this genus.

Monodictys capensis R.C. Sinclair et al., Mycotaxon 59: 359. 1996. Fig. 50.

Fig. 50.

Monodictys capensis. A, B. Colonies on rice straw in culture; C. Conidiophores and conidia; D, E. Conidia (E. bleached conidium). All from culture HR 1. Scale bars: A = 2 mm; B = 500 μm; C = 100 μm; D, E = 10 μm.

Specimen examined: Russia, St. Petersburg, Botanical garden of the Komarov Botanical Institute of the Russian Academy of Sciences, on dead wood of Padus avium, 19 Oct. 2005, V. Mel'nik, HR 1 = HHUF 29712 = LE 226298, culture CBS 134928 = VKM F-4506.

Notes: This material has been reported as M. capensis by Mel’nik & Shabunin (2010). In culture, abundant conidia similar to those on the natural host were produced on rice straw agar (RSA).

Discussion

New familial lineages

The Parabambusicolaceae is erected here to accommodate Aquastroma (Fig. 40) and Parabambusicola (Fig. 41), as well as two unnamed “Monodictys” species. This family is superficially similar to the Bambusicolaceae in having depressed globose to hemispherical ascomata. In particular, Parabambusicola, the type genus of the Parabambusicolaceae, has a bambusicolous habitat like that of Bambusicola (Dai et al. 2012). However, members of the Parabambusicolaceae have ascomata surrounded by stromatic tissue (Parabambusicola) or compressed necks with wide ostioles (Aquastroma), and also multiseptate, clavate to fusiform, hyaline ascospores, unlike those of the Bambusicolaceae. Asexual morphs possessing sporodochial conidiomata and muriform conidia like those of Monodictys are not found in the Bambusicolaceae.

A new family Sulcatisporaceae is introduced to encompas Magnicamarosporium and Sulcatispora genera nova, as well as Neobambusicola (Crous et al. 2014b). These three genera form a strongly supported clade (97 %), which is a sister of the Bambusicolaceae. The sexual morphs of species in the Sulcatisporaceae are superficially similar to those of Bambusicolaceae, but subglobose to obovoid conidia with muriform septation (Magnicamarosporium) or 1 to several septate conidia with or without striate ornamentation (Neobambusicola, Sulcatispora) are not found in species of the Bambusicolaceae.

We resurrect the Periconiaceae as a sister taxon of the Massarinaceae (Fig. 1). The name of “Periconieae” was originally used for dematiaceous hyphomycetes that have macronematous conidiophores and 1-celled, pigmented conidia forming conidial heads (Saccardo 1886). Later, the group was raised to familial rank, and Periconia and Stachybotrys were assigned to the Periconiaceae (Nannizzi 1934), the latter genus now being placed in its own family, Stachybotryaceae (Crous et al. 2014a). The Periconiaceae has long been ignored in modern fungal systematics, but it should be regarded as a natural taxon in the suborder Massarineae. The main genus Periconia has been treated as a member of the Massarinaceae based on a topology of genealogical trees (Kodsueb et al., 2007, Schoch et al., 2009, Zhang et al., 2009b, Zhang et al., 2009c, Zhang et al., 2012, Hyde et al., 2013). Species of Periconia, however, are separated from the Massarinaceae and form a strongly supported clade of the Periconiaceae (100 %; Fig. 1). Sexual morphs within the Periconiaceae (Fig. 42, Fig. 43) differ from those of Helminthosporium (Fig. 25) and Massarina (Fig. 26) in the Massarinaceae. Members in the latter family possess medium to large subglobose ascomata, which are covered by a clypeus, and have clavate asci. The small ascomata of Periconia superficially resemble those of Stagonospora (= Saccharicola, Massarinaceae; Fig. 27, Fig. 28, Fig. 29) but differ in having a peridium composed of small-sized compressed cells. The most diagnostic features of the Periconiaceae are the asexual morphs of Periconia; these have macronematous, mononematous conidiophores with globose to cylindrical, blastic conidia (Mason and Ellis, 1953, Ellis, 1971).

Dictyosporium and phenotypically similar genera, such as Aquaticheirospora and Pseudodictyosporium, have been considered to belong in the Massarinaceae (Wijayawardene et al. 2012). However, the group including dictyosporium-like fungi forms a distinct lineage (as Dictyosporiaceae nom. prov.; see Liu et al. 2015), independent from the Massarinaceae (Fig. 1). One of the diagnostic features of the Dictyosporiaceae is their multicellular cheiroid conidia (Fig. 2), and these morphological features separate it from other families in the Massarineae. Sexual morphs observed in the Dictyosporiaceae (Gregarithecium and Pseudocoleophoma; Fig. 3, Fig. 4, Fig. 5) are somewhat similar to those of the Massarinaceae (e.g., Stagonospora; Fig. 27, Fig. 28, Fig. 29) or the Periconiaceae (e.g., Periconia; Fig. 42, Fig. 43), but can be differentiated by their subglobose to hemispherical ascomata with or without surrounding stromatic tissue, short ascomatal necks without clypeus, cylindrical asci with a short stipes, and narrowly fusiform, 1-septate, hyaline ascospores.

Summary of accepted families and genera incertae sedis of the Massarineae

Based on our phylogenetic analyses and morphological evaluation of fungi in the Massarineae, we recognise the following 12 families, as well as “Massarineae, incertae sedis”.

Bambusicolaceae. This family was established by Hyde et al. (2013) to accommodate Bambusicola species on bamboo (Dai et al. 2012). Palmiascoma on dead fronds of palms in Thailand was also reported as a member of Bambusicolaceae (Liu et al. 2015).

Dictyosporiaceae (Fig. 2, Fig. 3, Fig. 4, Fig. 5). Dictyosporiaceae (nom. prov., see Liu et al. 2015) includes the four asexual genera Aquaticheirospora (Kodsueb et al. 2007), Dendryphiella (Jones et al. 2008), Dictyosporium (Fig. 2) and Pseudodictyosporium (Kirschner et al. 2013), and two new genera with sexual morphs, Gregarithecium (Fig. 3) and Pseudocoleophoma (Fig. 4, Fig. 5). The two asexual monotypic genera Cheirosporium (Cai et al. 2008) and Kamatia (Kirschner et al. 2013) also belong in the Dictyosporiaceae. “Diplococcium” asperum (Pirozynski, 1972, Goh and Hyde, 1998) is located in the Dictyosporiaceae (Fig. 1), but the type species of Diplococcium (D. spicatum) has phylogenetic affinity with the Helotiales (Shenoy et al. 2010). This species as well as Diplococcium pulneyense, the asexual morph of Otthia pulneyensis (Subramanian & Sekar 1987), are both related to the Dothideomycetes, and should be excluded from Diplococcium s. str. Likewise, “Paraconiothyrium” flavescens in the Dictyosporiaceae clade should be renamed, because the type lineage of the genus, represented by P. estuarinum, groups with the Didymosphaeriaceae (= Montagnulaceae) (Verkley et al. 2014). The Dictyosporiaceae clade includes Digitodesmium bambusicola (Cai et al. 2002), but the phylogenetic placement of the generic type D. elegans is unknown at present.

Didymosphaeriaceae (Fig. 6, Fig. 7, Fig. 8, Fig. 9, Fig. 10, Fig. 11). Ariyawansa et al. (2014) accepted 16 genera including Karstenula (Fig. 6), Neokalmusia (Fig. 7, Fig. 8), and Paraphaeosphaeria (Fig. 9) as members of the Didymosphaeriaceae, and subsequently three genera, Paracamarosporium, Pseudocamarosporium, and Pseudotrichia were also added to this family (Thambugala et al., 2014, Wijayawardene et al., 2014d). We assigned the hyphomycetous genus Spegazzinia (Fig. 10, Fig. 11) with basauxic conidiogenesis and pigmented conidia with spine-like appendages, to the Didymosphaeriaceae, although it has long been treated as “Ascomycota, genera incertae sedis” (Wijayawardene et al. 2012). However, taxonomic reassessment will be required for several genera, such as Neokalmusia, Paraconiothyrium, Paraphaeosphaeria and Pseudocamarosporium, because their monophyletic status was not supported in this study. In addition to these genera, the type species of Cucurbidothis (C. pityophila) resides in the Didymosphaeriaceae (Fig. 1). Phaeodothis winteri with a didymosphaeria-like sexual morph (Aptroot 1995), and Sporidesmiella fusiformis with macronematous conidiophores and obclavate, multi-distoseptate, pigmented conidia (Wu & Zhuang 2005) are placed in the Didymosphaeriaceae, but sequences of the type species of these genera are presently unknown. Two species of “Camarosporium” with muriform conidia, C. brabeji and C. leucadendri (Marincowitz et al. 2008), are positioned in the Didymosphaeriaceae, and have subsequently been allocated to Pseudocamarosporium and Paracamarosporium, respectively (Crous et al. 2015b). In contrast, the type species of Camarosporium (C. quaternatum) is known to cluster within the Pleosporineae (Crous et al., 2006, Wijayawardene et al., 2014b). Munkovalsaria appendiculata (Aptroot 2004) groups with Montagnula species (M. aloes, M. opulenta and M. graminicola). However, Munkovalsaria is apparently polyphyletic, because M. rubra is found outside the Pleosporales (Voglmayr and Jaklitsch, 2011, Hernández-Restrepo et al., 2014). Although Munkovalsaria is regarded as a member of the Dacampiaceae (Hyde et al. 2013), molecular data from the generic type (M. donacina) are needed to clarify its phylogenetic position.

Latoruaceae. This family was established by Crous et al. (2015a) to accommodate two hyphomycetous genera, Latorua and Polyschema, both mostly known from soil (Ellis, 1976, Shenoy et al., 2010, Crous et al., 2015a). More recently, a new genus Matsushimamyces from soil has been added to this family (Sharma et al. 2015).

Lentitheciaceae (Fig. 12, Fig. 13, Fig. 14, Fig. 15, Fig. 16, Fig. 17, Fig. 18, Fig. 19, Fig. 20, Fig. 21, Fig. 22, Fig. 23, Fig. 24). Darksidea (Knapp et al. 2015), Katumotoa (Fig. 12), Keissleriella (Fig. 13, Fig. 14, Fig. 15, Fig. 16, Fig. 17, Fig. 18), Lentithecium (Fig. 19, Fig. 20), Murilentithecium (Wanasinghe et al. 2014), Phragmocamarosporium (Wijayawardene et al. 2015), Poaceascoma (Phookamsak et al. 2015) and Tingoldiago (Fig. 24) have been assigned in the Lentitheciaceae, and we add another two genera, i.e., Neoophiosphaerella (Fig. 21) and Setoseptoria (Fig. 22, Fig. 23). The clade comprising species of these six genera received moderate BP support (71 %; Fig. 1). “Wettsteinina” lacustris is also a member of this clade (Schoch et al. 2009). In our preliminary analysis using SSU and LSU sequences, Ascorhombispora aquatica (Cai & Hyde 2007b) resided in the Lentitheciaceae (data not shown), but the phylogenetic affinity of this species with members in the Dictyosporiaceae (Cai & Hyde 2007b) or Didymosphaeriaceae (Shearer et al. 2009) has also been suggested. Further investigation using additional sequence data is needed to confirm familial placement of this genus.

Macrodiplodiopsidaceae. Three species with coelomycetous asexual morphs, i.e., Camarographium koreanum (on Cornus, Verkley et al. 2005), Macrodiplodiopsis desmazieri (on Platanus, Barr 1982), and Pseudochaetosphaeronema larense (a human pathogen, Ahmed et al. 2014) comprise the Macrodiplodiopsidaceae, which was recently established by Crous et al. (2015a).

Massarinaceae (Fig. 25, Fig. 26, Fig. 27, Fig. 28, Fig. 29). Byssothecium (Boise 1983), Helminthosporium (Fig. 25), Massarina (Fig. 26), Stagonospora (Fig. 27, Fig. 28, Fig. 29) and Suttonomyces (Wijayawardene et al. 2015) are accepted in the Massarinaceae (Fig. 1). Neottiosporina paspali (Sutton & Alcorn 1974) should be treated as Stagonospora paspali (Atkinson 1897) based on the phylogenetic topology (Fig. 1) as well as the morphological resemblance with S. paludosa, the type species of Stagonospora (Quaedvlieg et al. 2013). The two Corynespora species C. leucadendri (Quaedvlieg et al. 2013) and C. olivacea (Ellis 1960) clustered in this family but are not congeneric. The type species of Corynespora (C. mazei = C. cassiicola, Wei 1950) is sister of the clade of the Pleosporineae + Massarineae (Hyde et al. 2013), hence the Corynespora species in the Massarinaceae should be renamed. “Didymosphaeria” spartii has been previously transferred to Montagnula (Aptroot 1995) and also to Didymosphaerella (Chlebicki 2009), but these generic placements are inappropriate (Fig. 1). A cultural study of D. spartii (Scheinpflug 1958, using CBS 183.58) indicated that it has a dendrophoma-like asexual morph with conidia 4–7 × 1–2 μm in size.

Morosphaeriaceae (Fig. 30, Fig. 31, Fig. 32, Fig. 33, Fig. 34, Fig. 35, Fig. 36, Fig. 37, Fig. 38, Fig. 39). In addition to Aquilomyces (Fig. 30), Helicascus (Fig. 35, Fig. 36, Fig. 37) and Morosphaeria (Fig. 38, Fig. 39), which were previously recognised as members of the Morosphaeriaceae (Suetrong et al., 2009, Hyde et al., 2013, Knapp et al., 2015), we add the new genus Clypeoloculus (Fig. 31, Fig. 32, Fig. 33, Fig. 34) to this family. Species in these four genera are only known to have sexual morphs. A pleurophomopsis-like conidial state has been reported for H. aquaticus (Zhang et al. 2013) as its asexual morph, and we also observed this for H. aquaticus, as well as for four species in Clypeoloculus, but these should be regarded as spermatial morphs. “Pithomyces” valparadisiacus (Kirk, 1983, Marincowitz et al., 2008) is basal to Clypeoloculus species, and is found to be a rare case in which a species known only from an asexual form belongs to the Morosphaeriaceae (Fig. 1). A hyphomycetous genus Pithomyces is regarded as a heterogenous group comprising more than three lineages (Phookamsak et al. 2013), and the generic type (P. flavus) has phylogenetic affinity with Astrosphaeriella basal to family Aigialaceae (Pratibha & Prabhugaonkar 2015).

Parabambusicolaceae (Fig. 40, Fig. 41). This new family includes two new sexual genera, Aquastroma (Fig. 40) and Parabambusicola (Fig. 41). Multiseptospora formerly classified in Pleosporales, genera incertae sedis (Liu et al. 2015) is also accepted as a member of the Parabambusicolaceae. Two unnamed Monodictys species with muriform conidia are also located in this clade, but this genus is obviously heterogenous (see Notes in Monodictys capensis).

Periconiaceae (Fig. 42, Fig. 43). Bambusistroma (Adamčík et al. 2015), Flavomyces (Knapp et al. 2015), Periconia (Fig. 42, Fig. 43) and Noosia (Crous et al. 2011a) are accepted in this family. “Sporidesmium” tengii is also placed in the Periconiaceae, but the phylogenetic position of the type species of Sporidesmium (S. atrum) is currently unknown. According to Shenoy et al. (2006), this genus is not monophyletic and species in Sporidesmium are phylogenetically distributed in seven lineages between two major ascomycete classes, Dothideomycetes and Sordariomycetes. Periconia species used in this study did not form a single clade. Therefore, a taxonomic revision of Periconia species along with Noosia and “Sporidesmium” tengii should be conducted in future.

Sulcatisporaceae (Fig. 44, Fig. 45, Fig. 46). Magnicamarosporium (Fig. 44) on Diplospora (Rubiaceae) and two species of Sulcatispora, S. acerina (Fig. 45) on Acer (Aceraceae) and S. berchemiae (Fig. 46) on Berchemia (Rhamnaceae), are assigned to the Sulcatisporaceae. Neobambusicola (on Grewia sp.; Malvaceae) previously placed in the Bambusicolaceae (Crous et al. 2014b) is accepted here as a member of the Sulcatisporaceae.

Trematosphaeriaceae (Fig. 47). This family was established by Suetrong et al. (2011b) to accommodate the three genera, Falciformispora, Halomassarina, and Trematosphaeria (Fig. 47) (Hyde et al. 2013). Later, Bryosphaeria, Hadrospora, and Medicopsis were listed as additional members of the Trematosphaeriaceae (Wijayawardene et al. 2014c). However, the placement of Bryosphaeria has not been verified by molecular data. Hadrospora may have affinity with the Lindgomycetaceae (Tanaka, unpublished data), and Medicopsis belongs to the Pleosporineae, rather than the Massarineae (Ahmed et al. 2014). A coelomycetous species, Amarenographium solium with pigmented muriform conidia (Hodhod et al. 2012), has been reported to have phylogenetic affinity to the Trematosphaeriaceae/Didymosphaeriaceae, but molecular data from the generic type (A. metableticum, Eriksson 1982) are currently unavailable.

Massarineae incertae sedis (Unknown Clades I–V; Fig. 48, Fig. 49, Fig. 50). Fuscostagonospora (Fig. 48) on bamboo is located in the Unknown Clade I and its familial placement remains unresolved. Pseudoxylomyces (Fig. 49), from freshwater environments, occupies a basal position in the Trematosphaeriaceae (Unknown Clade II, Fig. 1). Bactrodesmium cubense (Unknown Clade III), a sporodochial fungus with pigmented phragmoconidia (Zucconi & Lunghini 1997), is a sister taxon of the Morosphaeriaceae (Fig. 1). Monodictys capensis (Fig. 50), without a known sexual morph (Mel’nik & Shabunin 2010), and Inflatispora pseudostromatica, without a known asexual morph (Zhang et al. 2011), groups in Unknown Clade IV.

Morphological characteristics of sexual morphs

The Massarinaceae clade is supported by high BP support (100 %, Fig. 1), but it is somewhat difficult to circumscribe based on their sexual morphs. The Massarinaceae mainly contains two types of sexual morphs; massarina-like and stagonospora-like. The sexual morph observed in Helminthosporium (Fig. 25) and Massarina (Fig. 26) have immersed, hemispherical, medium to large (ca. 400–700 μm diam) ascomata with short papillate necks covered by a clypeus, clavate asci, and broadly fusiform to ellipsoidal, 1- to 3-septate, hyaline ascospores surrounded by a thick conspicuous sheath. Those of Stagonospora (Fig. 27, Fig. 28, Fig. 29) are characterised by globose, small (ca. 200–300 μm diam) ascomata lacking a clypeus, cylindrical to clavate asci, and fusiform, 1-septate, hyaline ascospores with a thin sheath. This family, however, includes further sexual species, such as “Didymosphaerella (or Montagnula)” spartii with globose ascomata and thick-walled, 1-septate, pigmented ascospores (Aptroot, 1995, Chlebicki, 2009), and Byssothecium circinans with subglobose ascomata with broadly papillate necks and versicoloured ascospores (Boise 1983). Further molecular investigation using additional taxa close to Massarina and Stagonospora may provide sufficient data to subdivide this family.

Bambusicolaceae, Macrodiplodiopsidaceae, Parabambusicolaceae and Sulcatisporaceae presently include only a few species. One to two genera with sexual morphs are recognised in each family: Bambusicola (Dai et al. 2012) and Palmiascoma (Liu et al. 2015) in the Bambusicolaceae, Macrodiplodiopsis (Crous et al. 2015a) in the Macrodiplodiopsidaceae, Aquastroma (Fig. 40) and Parabambusicola (Fig. 41) in the Parabambusicolaceae, and Sulcatispora (Fig. 45, Fig. 46) in the Sulcatisporaceae. The sexual morphs in the Bambusicolaceae, Parabambusicolaceae and Sulcatisporaceae are similar to those of Massarina s. lat. (Aptroot, 1998, Tanaka and Harada, 2003b) in having immersed to erumpent, hemispherical to depressed globose ascomata. Species in the Bambusicolaceae have conical ascomata with short ostiolar necks, narrow pseudoparaphyses, cylindrical asci, and narrowly fusiform, 1-septate ascospores, whereas those in the Parabambusicolaceae lack a prominent neck, and have relatively wide pseudoparaphyses, clavate to broadly cylindrical asci, and clavate to fusiform ascospores with multiple transverse septa. Species in the Sulcatisporaceae have subglobose ascomata, trabeculate pseudoparaphyses, clavate asci, and broadly fusiform ascospores, and occur on woody host plant rather than herbaceous host. On the other hand, the sexual morphs in the Macrodiplodiopsidaceae are similar to those of Pleomassariaceae s. lat. (Barr 1982). Macrodiplodiopsis desmazieri (= Splanchnonema platani) in this family is characterised by large-sized ascomata (500–900 μm diam) with thick ascomatal wall, clavate asci, and dark brown ascospores with 3–5(–6) eudistosepta (Barr, 1982, Crous et al., 2015a).

Morosphaeriaceae (Fig. 30, Fig. 31, Fig. 32, Fig. 33, Fig. 34, Fig. 35, Fig. 36, Fig. 37, Fig. 38, Fig. 39) and Trematosphaeriaceae (Fig. 47), mostly found from aquatic environments, are characterised by immersed to erumpent, medium to large, black, carbonaceous ascomata with or without papillate necks, but necks are prominent and/or surrounded by a clypeus in members of the Morosphaeriaceae. Asexual morphs are relatively rare in these families with few exceptions, i.e., Pithomyces valparadisiacus (Morosphaeriaceae, Kirk 1983), and Trematosphaeria grisea (Trematosphaeriaceae, Ahmed et al. 2014). This tendency has been observed in the Aliquandostipitaceae (Jahnulales), which also occurs in aquatic habitats, and a few asexual species in the genera Brachiosphaera, Speiropsis, and Xylomyces (Suetrong et al. 2011a).

We tentatively accept the Lentitheciaceae as a diverse assemblage. Four groups seem to be recognised based on morphological variations in the sexual morphs: Lentithecium with immersed, globose ascomata without clypeus, and broadly fusiform, 1-septate, hyaline ascospores (Fig. 19, Fig. 20), Keissleriella with small, globose to subglobose ascomata with setose necks and several septate ascospores (Fig. 13, Fig. 14, Fig. 15, Fig. 16, Fig. 17, Fig. 18), Setoseptoria (Fig. 22, Fig. 23) and Tingoldiago (Fig. 24) with single to grouped, hemispherical ascomata and cylindrical to fusiform ascospores, Katumotoa (Fig. 12) and Neoophiosphaerella (Fig. 21) with subglobose ascomata without prominent necks and fusiform or filiform ascospores. The morphological diversity of the family, as well as relatively low phylogenetic support of the clade (71 %, Fig. 1), strongly suggests polyphyly of the Lentitheciaceae s. lat. used in this study.

Similarly, various sexual morphs are observed in the Didymosphaeriaceae (Fig. 6, Fig. 7, Fig. 8, Fig. 9). Most of the species in this family have previously been placed in several genera of the Pleosporineae, which include Didymosphaeria (Montagnula opulenta, Phaeodothis winteri), Leptosphaeria (Paraconiothyrium fuckelii, Neokalmusia scabrispora), and Phaeosphaeria (Neokalmusia brevispora). The type species of Montagnula (M. infernalis; Berlese 1896) has been treated as Leptosphaeria or Pleospora (Wehmeyer 1961). Species having muriform ascospores similar to Pleospora are known in Deniquelata (Ariyawansa et al. 2013) and Tremateia (Kohlmeyer et al. 1995). Sexual morphs in the Didymosphaeriaceae have small to medium, globose to subglobose ascomata with papillate necks and melanised, didymo/phragmo/dictyo-ascospores resembling those of the Pleosporineae, but the ascomata are surrounded by abundant hyphae that form an apical clypeus at times, as was defined by Barr (2001). In addition, Bimuria (Hawksworth et al. 1979), a soil-borne ascomycete, with unique morphological characters such as ascomata composed of very thin peridium, 2-spored asci, and muriform, dark brown, verrucose ascospores, also belongs in the Didymosphaeriaceae. The family includes a wide variety of sexual morphs with dark coloured ascospores, as well as coniothyrium-like or Spegazzinia asexual morphs, and the clade receives 99 % BP support (Fig. 1).

In contrast, the Dictyosporiaceae and Periconiaceae mostly comprise asexual species. The few known sexual morphs in these families are characterised by single to grouped, globose to hemispherical ascomata with or without surrounding stromatic tissue, cylindrical asci with short stipes, and fusiform to broadly fusiform, 1-septate, hyaline ascospores. Examples are Dictyosporium (e.g., D. meiosporum; Liu et al. 2015), Gregarithecium (Fig. 3) and Pseudocoleophoma (Fig. 4, Fig. 5) in the Dictyosporiaceae and Bambusistroma (Adamčík et al. 2015) and Periconia (Fig. 42, Fig. 43) in the Periconiaceae. These morphological features agree in most respects with those of Massarina s. lat. (Aptroot, 1998, Tanaka and Harada, 2003b) and are similar to those of species in the Massarinaceae (e.g., Stagonospora). In the Latoruaceae, no sexual morphs are known at present (Crous et al. 2015a).

Morphological characteristics of asexual morphs

Although various asexual genera are found in the Massarineae, hyphomycetous species with macronematous, mononematous conidiophores are restricted to the Massarinaceae [Helminthosporium (Fig. 25) and Corynespora], Periconiaceae [Periconia (Fig. 43) and Sporidesmium], Dictyosporiaceae (Diplococcium and Dendryphiella), and Didymosphaeriaceae (Sporidesmiella). Similarly, synnematous hyphomycetes are extremely rare in this suborder and have only been observed in one instance, in Aquaticheirospora (Kodsueb et al. 2007) in the Dictyosporiaceae. Hyphomycetous asexual morphs without prominent conidiophores are found in the Periconiaceae (Noosia), Latoruaceae (Latorua and Polyschema), and the Morosphaeriaceae (Pithomyces). Sporodochial hyphomycetes are scattered through the Dictyosporiaceae [Dictyosporium (Fig. 2) and its morphologically similar genera], Didymosphaeriaceae (Spegazzinia, Fig. 10, Fig. 11), Parabambusicolaceae (Monodictys), and Unknown Clades III (Bactrodesmium) and IV (Monodictys, Fig. 50). In contrast, coelomycetous genera seem to be more common than hyphomycetous asexual morphs and are present throughout this suborder, i.e., in the Bambusicolaceae (Bambusicola), Dictyosporiaceae (Pseudocoleophoma, Fig. 4, Fig. 5), Didymosphaeriaceae [Paracamarosporium, Paraconiothyrium, Pseudocamarosporium, Karstenula, Paraphaeosphaeria (Fig. 9)], Lentitheciaceae [Keissleriella (Fig. 13, Fig. 16), Setoseptoria], Macrodiplodiopsidaceae (Camarographium, Macrodiplodiopsis, Pseudochaetosphaeronema), Massarinaceae (Stagonospora, Fig. 27, Fig. 28, Fig. 29), Sulcatisporaceae [Magnicamarosporium (Fig. 44), Sulcatispora (Fig. 45, Fig. 46)] and Unknown Clades I (Fuscostagonospora, Fig. 48).

In general, Dictyosporiaceae is characterised by hyphomycetous asexual morphs with pigmented cheiroconidia. These usually have sporodochial (e.g., Dictyosporium, Digitodesmium, and Pseudodictyosporium) or synnematous conidiomata (e.g., Aquaticheirospora). Dendryphiella and Diplococcium have macronematous, mononematous conidiophores with pigmented, multi-celled, tretic conidia. The coelomycetous morphs known in this family are “Paraconiothyrium” flavescens and Pseudocoleophoma species, with phialidic, 1-celled conidia. In contrast, most asexual species in the Periconiaceae have mononematous, macronematous conidiophores and produce conidia in chains (e.g., Periconia), but some deviating species, such as “Sporidesmium” tengii have monoblastic, obpyriform, multi-septate, pigmented conidia (Wu & Zhuang 2005), and Noosia which lacks a prominent conidiophore (Crous et al. 2011a), are also known in the family. Further phylogenetic investigations of asexual genera exhibiting morphological resemblance to these two groups but without molecular information, such as dictyosporium-like (e.g., Cheiromycella, Digitomyces, Paratetraploa; Seifert et al. 2011) and periconia-like genera (e.g., Lacellinopsis, Sadasivania, Trichobotrys; Seifert et al. 2011), should be conducted to evaluate the phylogenetic significance of these phenotypic characters at familial level, and/or to clarify taxonomic understanding of the Dictyosporiaceae and Periconiaceae. Although in some families (e.g., the Didymosphaeriaceae and Massarinaceae), asexual morphs are extremely diverse and thus asexual characters alone appear insufficient for familial circumscriptions, there are many recent examples of families that are well defined by asexual morphological characters along with their phylogenetic information. These include the Coniothyriaceae (De Gruyter et al. 2013), Cladosporiaceae (Bensch et al. 2012), Kirschsteiniotheliaceae (Boonmee et al. 2012), Planistromellaceae (Minnis et al. 2012), and Tetraplosphaeriaceae (Tanaka et al. 2009).

Habitat

In contrast to the Pleosporineae which comprises numerous plant pathogens (Zhang et al. 2009b), such as necrotrophs (e.g., Alternaria, Bipolaris) and hemibiotrophs (e.g., Leptosphaeria, Setosphaeria) on economically important crops (Ohm et al. 2012), the Massarineae, a sister lineage of the Pleosporineae, mostly includes saprobes on various plant substrates.

Species in the Morosphaeriaceae and Trematosphaeriaceae have been reported mostly as saprobes on decomposed woody substrates submerged in freshwater or marine habitats. Recently, several human pathogenic species have been reported in genera in the Trematosphaeriaceae, and an association of the virulence factors with oligotrophism or halotolerance has been suggested (Ahmed et al. 2014). This may further indicate the presence of undescribed lineages of mycetoma agents in the Morosphaeriaceae.

Most other families in the Massarineae do not seem to have specific habitat preferences in each family. Species in the Dictyosporiaceae, for example, are reported from ecologically diverse environments, i.e., terrestrial monocots and dicots (e.g., Dictyosporium strelitziae and Diplococcium asperum; Pirozynski, 1972, Crous et al., 2009a), submerged wood in freshwater (e.g., Aquaticheirospora lignicola; Kodsueb et al. 2007), mangroves or drift wood in marine ecosystems (Dictyosporium inflatum; Kirschner et al. 2013), and rhizosphere soil (Pseudodictyosporium elegans; De Gruyter et al. 2013). In the Didymosphaeriaceae, many coelomycetous species similar to Phoma s. lat. are generally soil-borne fungi (Verkley et al. 2014), but species with other ecological features, e.g., endophytes (Dendrothyrium; Verkley et al. 2014), mycoparasites (Paraphaeosphaeria minitans; Campbell 1947), symbionts with scale insects (Cucurbidothis pityophila; Casagrande, 1969, Barr, 1990b), and marine saprobes (Tremateia halophila; Kohlmeyer et al. 1995) are also known. Paraconiothyrium fuckelii in the Didymosphaeriaceae is known as an agent of mycetoma and a serious plant pathogen of the Rosaceae (Verkley et al. 2014).

In the suborder Massarineae, relatively few species are known as plant pathogens. They are restricted to the Didymosphaeriaceae (Paraconiothyrium tiliae, Deniquelata barringtoniae; Butin and Kehr, 1995, Ariyawansa et al., 2013), Massarineae (Byssothecium circinans, Stagonospora spp., Helminthosporium solani; Kaiser et al., 1979, Semeniuk, 1983, Errampalli et al., 2001) and Periconiaceae (Periconia igniaria; Kolomiets et al. 2008); lineages rich in asexual species. Coprophilous species such as those in the Sporormiaceae (Kruys & Wedin 2009) and lichenicolous species such as Arthopyrenia salicis (Nelsen et al. 2009) are not known from the Massarineae.

Future studies

We examined the morphology of 106 taxa belonging to the Massarineae and analysed their phylogenetic relationships based on sequences from SSU rDNA, LSU rDNA and tef1, along with sequences of 131 taxa previously assigned to the Massarineae. Our results delineated 10 new genera and 29 new species/new combinations in more than 12 families and five unknown lineages (Fig. 1). Our study has contributed to the understanding of species diversity within the Massarineae, and improves the classification of these species, but several taxonomic issues remain unclear. The Lentitheciaceae s. lat. used here, for example, is probably polyphyletic based on the ecological and morphological divergence of these species, as well as based on our phylogenetic analysis, although we have tentatively accepted the family at this point. Furthermore, the presence of several lineages (Unknown Clades I to V), which we were unable to assign to any existing families, makes it certain that there are many undiscovered taxa, which should form a new family in the suborder.

Several comprehensive works have been published recently that have shown interest in the phylogeny of the Dothideomycetes. Schoch et al. (2006) provided a fundamental overview of the class, and its composition of two subclasses, the Pleosporomycetidae (pseudoparaphyses present) and the Dothideomycetidae (pseudoparaphyses absent), based on phylogenetic analyses of four loci (nucSSU, nucLSU rDNA, tef1, rpb2) from 96 taxa. Schoch et al. (2009) reconstructed the dothideomycete phylogeny based on five genes (nucSSU, nucLSU rDNA, tef1, rpb1, rpb2) from 356 isolates, and discussed the evolutionary transitions of ecological characteristics. Following these publications, the understanding of the natural relationships among dothideomycetous taxa and their taxonomic revision have tremendously progressed, and the number of accepted families in this class has been increased from 41 (Schoch et al. 2009) to 105 (Hyde et al. 2013) with or without molecular evidence. Monographic revision based on the type specimens along with phylogeneric analyses have been published by Zhang et al. (2012) who concentrated on reassessment of genera in the Pleosporales, and by Hyde et al. (2013) who circumscribed each family in the Dothideomycetes. The phylogenies of plant pathogenic taxa in this class, such as Alternaria (Woudenberg et al. 2013), Cercospora (Groenewald et al. 2013), Cladosporium (Bensch et al. 2012), Phoma (De Gruyter et al. 2013), Pseudocercospora (Crous et al. 2013a), Septoria (Verkley et al., 2013, Quaedvlieg et al., 2013) and several genera in the Botryosphaeriales (Wikee et al., 2013, Slippers et al., 2013, Phillips et al., 2013), have been intensively studied. More recently, a special issue of Phytotaxa has been published, comprising 26 articles focused on the taxonomy and phylogeny of the Dothideomycetes (Mckenzie et al. 2014).

However, these works on Dothideomycetes include relatively few species belonging to the Massarineae. The number of Massarineae taxa used in phylogenetic analyses of the Dothideomycetes was only six (among 96 taxa; Schoch et al. 2006), and 35 (among 356 taxa; Schoch et al. 2009). In the revision of the Pleosporales by Zhang et al. (2012) who recognised the validity of Massarineae, only 46 taxa of the suborder were phylogenetically analysed. Furthermore, most families in this suborder including the Bambusicolaceae (Hyde et al. 2013), Lentitheciaceae (Zhang et al. 2009b), Morosphaeriaceae (Suetrong et al. 2009), and Trematosphaeriaceae (Suetrong et al. 2011b), have been lineages recently recognised based on molecular data, and are currently characterised morphologically by only a few genera. To provide more precise circumscription of these families, as well as of the unnamed lineages we have found (Clades I to V, Fig. 1), further phylogenetic analyses using a greater number of taxa should be conducted with priority. These should include a fundamental taxonomic approach to species discovery along with sequence verification, such as the work undertaken by Fungal Planet (Crous et al. 2015c), Fungal Diversity Notes (Liu et al. 2015) and Fungal Systematics and Evolution (FUSE) (Crous et al. 2015b).

In due course further phylogenetic study is also necessary of the more than 1 500 asexual genera treated as “Ascomycota, genera incertae sedis” (Wijayawardene et al. 2012); those without sexual links or molecular phylogenetic information. However, asexual characters alone may not provide good resolution for familial circumscription for some aquatic lineages, such as Morosphaeriaceae and Trematosphaeriaceae, in which predominantly sexual species are known. In contrast, species in the Dictyosporiaceae and Periconiaceae comprise a high proportion of asexual taxa. These have previously been treated as Massarinaceae (Hyde et al. 2013), but we have further characterised these as independent families based on their sexual morphs, in addition to their distinct asexual morphologies. Recognition of the Dictyosporiaceae and Periconiaceae partially indicates the phylogenetic significance of asexual taxa, but further comprehensive taxonomic work based on the holomorph and not weighted towards a particular fungal morph (e.g., Crous et al., 2009b, Tanaka et al., 2009, Tanaka et al., 2010, Voglmayr and Jaklitsch, 2011, Dai et al., 2012, Phillips et al., 2013, Boonmee et al., 2014) should be conducted to revise sexual morph-based fungal systematics.