Towards a natural classification and backbone tree for ...

Fungal Diversity (2015) 74:199–266 DOI 10.1007/s13225-015-0348-3

Towards a natural classification and backbone tree for Lophiostomataceae, Floricolaceae, and Amorosiaceae fam. nov. Kasun M. Thambugala 1,2,3 & Kevin D. Hyde 2,3,4 & Kazuaki Tanaka 5 & Qing Tian 2,3 & Dhanushka N. Wanasinghe 2,3,4 & Hiran A. Ariyawansa 1,2,3 & Subashini C. Jayasiri 2,3 & Saranyaphat Boonmee 2 & Erio Camporesi 9,10 & Akira Hashimoto 5,6 & Kazuyuki Hirayama 7 & René K. Schumacher 11 & Itthayakorn Promputtha 8 & Zuo-Yi Liu 1

Received: 13 July 2015 / Accepted: 10 September 2015 / Published online: 3 November 2015 # School of Science 2015

Abstract The genera Lophiostoma, Misturatosphaeria and several other allied taxa in Lophiostomataceae are revisited. Accounts of these taxa, including their history, morphology, and family placement, based on molecular phylogeny, are provided. Type or representative specimens of Lophiostoma and Misturatosphaeria were examined and fresh specimens were obtained from Germany, Italy, Japan and Thailand. A multigene phylogenetic analysis of the lophiostomataceous genera Floricola, Lophiostoma, Misturatosphaeria and related taxa is provided. Sixteen genera including Lophiostoma, Lophiohelichrysum, Dimorphiopsis, Platystomum and Vaginatispora, plus eleven newly introduced genera Biappendiculispora, Alpestrisphaeria, Capulatispora, Coelodictyosporium, Guttulispora, Lophiopoacea, Neotrematosphaeria, Paucispora, Pseudolophiostoma, Pseudoplatystomum and Sigarispora are accepted in Lophiostomataceae based on morphology and phylogeny.

Lophiostoma caulium, Lophiostoma arundinis and Lophiostoma caudatum are accommodated in Sigarispora. Lophiostoma winteri and Lophiostoma fuckelii are placed in the genera Lophiopoacea and Vaginatispora respectively. Three Curreya species and Misturatosphaeria claviformis are transferred to a new genus, Neocurreya. All other Misturatosphaeria species except Misturatosphaeria aurantiinotata and M. uniseptata are separated in the new genera Asymmetrispora, Aurantiascoma, Magnibotryascoma, Pseudoaurantiascoma and Pseudomisturatosphaeria based on their morphological and phylogenetic affinities. Another new genus, Ramusculicola is introduced for a new collection from Thailand. These seven new genera are accommodated in a new family Floricolaceae, together with Floricola and Misturatosphaeria. Several massarina-like species clustered as a sister clade to Amorosia littoralis and are accommodated in a new genus Angustimassarina. A new family Amorosiaceae is

* Zuo-Yi Liu [email protected]

5

Faculty of Agriculture and Life Sciences, Hirosaki University, 3 Bunkyo-cho Hirosaki, Aomori 036-8561, Japan

6

The United Graduate School of Agricultural Sciences, Iwate University, 18-8 Ueda 3 chome, Morioka 020-8550, Japan

7

Apple Experiment Station, Aomori Prefectural Agriculture and Forestry Research Center, 24 Fukutami, Botandaira, Kuroishi, Aomori 036-0332, Japan

8

Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand

9

A. M. B. Gruppo Micologico Forlivese BAntonio Cicognani^, Via Roma 18, Forlì, Italy

10

A.M.B. Circolo Micologico BGiovanni Carini^, C.P. 314 Brescia, Italy

11

Hölderlinsraße 25, 15517 Fürstenwalde/Spree, Germany

1

Guizhou Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang, Guizhou 550006, People’s Republic of China

2

Center of Excellence in Fungal Research, Chiang Rai 57100, Thailand

3

School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand

4

Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Science, Kunming 650201, Yunnan, China

200

Fungal Diversity (2015) 74:199–266

proposed to accommodate the genera Amorosia and Angustimassarina. The putatively named species Decaisnella formosa and Thyridaria macrostomoides form a separate clade together with a new genus Lignosphaeria which is placed in Dothideomycetes, genera incertae sedis.

of various woody plants and herbaceous plants in terrestrial and aquatic environments (Ellis and Ellis 1985; Mugambi and Huhndorf 2009; Zhang et al. 2009; Hirayama and Tanaka 2011; Hyde et al. 2013). Historic overview of Lophiostomataceae

Keywords Dothideomycetes . Floricola Lophiostoma . Misturatosphaeria . Multi-gene phylogeny new species new genus . Pleosporales taxonomy

Introduction Lophiostomataceae is an interesting and important family in the order Pleosporales, Dothideomycetes. Most species of this family are saprobic and occur mainly on twigs, stems or bark Table 1 study

The family Lophiostomataceae was introduced by Nitschke (1869) with Lophiostoma macrostomum (Tode) Ces. & De Not. as the type species (Eriksson 1981; Mugambi and Huhndorf 2009) and this family has been treated in different orders by various authors (Luttrell 1973; von Arx and Müller 1975; Barr 1987, 1992; Holm and Holm 1988; Hawksworth et al. 1995; Kirk et al. 2008; Zhang et al. 2012). Luttrell (1973) included Lophiostomataceae in Pleosporales with seven other families. Von Arx and Müller (1975) treated this family in the

Historical treatments of Lophiostomataceae by various authors and accepted genera in Amorosiaceae and Floricolaceae fam. nov. in this

von Arx and Holm and Müller 1975 Holm 1988

Barr (1992)

Hawksworth et al. 1995 Lumbsch and Huhndorf 2010

Hyde et al. 2013

Byssolophis Lophiella Lophiostoma Muroia

Lophiostoma Lophiotrema Massariosphaeria Navicella

Cilioplea Entodesmium Herpotrichia Lophiostoma

Cilioplea Entodesmium Herpotrichia Keissleriella

Byssolophis Cilioplea Epiphegia Lophiella

Lophiostoma Lophiostomataceae Misturatosphaeria Alpestrisphaeria Tumularia Biappendiculispora Capulatispora

Ostropella

Trematosphaeria

Lophiotrema

Lophiella

Lophionema

Coelodictyosporium

Lophiostoma Lophiotrema Massariosphaeria Misturatosphaeria Muroia

Dimorphiopsis Guttulispora Lophiohelichrysum Lophiopoacea Lophiostoma

Quintaria

Neotrematosphaeria Paucispora Platystomum

Platystomum Sampaioa

Massarina Lophionema Trichometasphaeria Lophiostoma Lophiotrema Massarina Massariosphaeria Muroia Quintaria Sampaioa Trichometasphaeria

Present study

Pseudolophiostoma Pseudoplatystomum Sigarispora Vaginatispora Amorosiaceae Amorosia Angustimassarina Floricolaceae Asymmetrispora Aurantiascoma Floricola Magnibotryascoma Misturatosphaeria Neocurreya Pseudoaurantiascoma Pseudomisturatosphaeria Ramusculicola


order Dothideales under the suborder Pseudosphaeriineae with seven other genera (Table 1). Barr (1987) synonymized Massarinaceae with Lophiostomataceae after removing melanommataceous taxa and accepted Cilioplea, Dangeardiella, Herpotrichia, Lophidiopsis, Lophiostoma, Massarina and Trichometasphaeria. Holm and Holm (1988) accepted taxa characterized by immersed-erumpent ascomata with a distinctly flattened neck and opening by slit-like ostioles, mostly clavate, bitunicate asci and 1-septate, multiseptate or muriform, hyaline to dark brown ascospores and accepted five genera. They treated Platystomaceae as a synonym of Lophiostomataceae, based on its short, blackish beak and periphysate ostiole. Barr (1992) included Entodesmium in Lophiostomataceae with six other North American genera. Hawksworth et al. (1995) included 14 genera in Lophiostomataceae and treated this family in Dothideales. Kirk et al. (2008) accepted 15 genera with 138 species in Lophiostomataceae. The family Lophiostomataceae was thought to be heterogeneous or paraphyletic, but recent phylogenetic analyses support the monophyletic status of Lophiostomataceae sensu stricto (Mugambi and Huhndorf 2009; Zhang et al. 2012; Hyde et al. 2013; Wijayawardene et al. 2014b). In their multi-gene analysis, Schoch et al. (2006, 2009) showed Lophiostomataceae to belong in Pleosporales. Mugambi and Huhndorf (2009) introduced a new genus Misturatosphaeria in Lophiostomataceae, that was supported by combined phylogenetic analyses of LSU and TEF sequence data and Misturatosphaeria formed a strongly supported sister clade to Lophiostoma. Lumbsch and Huhndorf (2010) included eleven genera in Lophiostomataceae, while Hyde et al. (2013) reduced this to three genera based on morphology and molecular data (Table 1). Morphological treatments of Lophiostomataceae followed the characters described by Holm and Holm (1988), however phylogenetic analyses have shown that the taxa with lophiostomataceous characters can be placed in several families outside of Lophiostomataceae (Hyde et al. 2002; Mugambi and Huhndorf 2009; Zhang et al. 2009; Hirayama and Tanaka 2011; Hyde et al. 2013). Zhang et al. (2009) showed that Lophiostoma, Lophiotrema and Massarina formed wellsupported subclades in Pleosporales, while recent studies (Zhang et al. 2012; Hyde et al. 2013; Quaedvlieg et al. 2013) confirm the placement of Massarina in Massarinaceae, Pleosporales. Hirayama and Tanaka (2011) introduced a new family Lophiotremataceae for Lophiotrema, which, is morphologically similar to Lophiostoma, but they are distinct lineages in phylogenetic analyses. Mugambi and Huhndorf (2009) showed that a putatively named strain of the type of Platystomum (Platystomum compressum (Pers.) Trevis.) is not phylogenetically related to Lophiostomataceae. Wijayawardene et al. (2014b) and Tian et al. (in prep) accepted Herpotrichia in Melanommataceae. Quintaria (another morphologically similar genus to Lophiostomataceae) was

201

introduced by Kohlmeyer and Volkmann-Kohlmeyer (1991) in Lophiostomataceae. Phylogenetic analysis of Schoch et al. (2009) and Suetrong et al. (2009), showed that the type of Quintaria (Q. lignatilis) forms a sister group to Testudinaceae, while Q. submersa clustered near Lindgomycetaceae (Schoch et al. 2009). Although the familial placement of Quintaria is not definitively known, it is placed apart from Lophiostomataceae (Suetrong et al. 2009). Wang et al. (2007) showed the polyphyletic nature of Massariosphaeria based on rDNA and RPB2 phylogenies. However, Massariosphaeria species needs further sampling with fresh collections, reference or ex-type strains and molecular data to obtain a natural classification. Thambugala et al. (2014a) transferred another lophiostomataceous genus, Dangeardiella to Pleosporales, genera incertae sedis considering the morphology and SSU phylogeny of D. macrospora (J. Schröt.) Sacc. & P. Syd. Byssolophis may be a synonym of Lophiostoma as it has slit-like ostioles, cylindrical asci and hyaline, 1-septate ascospores, as well as the form of pseudoparaphyses, which are similar to those of Lophiostoma (Zhang et al. 2012). Barr (1990) referred Navicella to the family Massariaceae based on the wide endotunica, thin apical ring and distoseptate ascospores. Ariyawansa et al. (2014a) however assigned Navicella to Melanommataceae considering the morphological similarities with Melanommataceae, such as the superficial globose to subglobose, coriaceous ascomata, long, trabeculate pseudoparaphyses, embedded in a gelatinous matrix and brown ascospores. Phookamsak et al. (2014) placed Entodesmium in Phaeosphaeriaceae, while Wijayawardene et al. (2014b) listed Cilioplea in Dothideomycetes, genera incertae sedis. Thambugala et al. (2014b) did not accept Lophiostomataceae as a member of Dothideales. Circumscription of Lophiostomataceae The family Lophiostomataceae likely includes numerous genera and species and recent phylogenetic studies have confirmed the generic placement of several taxa (Zhang et al. 2012; Hirayama and Tanaka 2011; Liu et al. 2015). Many taxa have lophiostoma-like characters, but their inclusion in Lophiostomataceae is uncertain (Mugambi and Huhndorf 2009; Zhang et al. 2009; Hirayama and Tanaka 2011). There is a lack of data for asexual morphs of Lophiostomataceae. Molecular data are therefore needed to properly link these asexual morphs to their sexual counterparts within the family. Mugambi and Huhndorf (2009) introduced Misturatosphaeria as a new genus in Lophiostomataceae based on morphology and combined LSU and EF1-α sequence data. Misturatosphaeria differs from Lophiostoma in having erumpent to superficial ascomata, with rounded apices that are often raised, rounded ostiolar openings plugged by gelatinous tissue and cylindrical to clavate asci with phragmo or dictyospores (Mugambi and Huhndorf 2009). In our phylogenetic analysis, clade C which comprises Curreya,

202

Floricola, Misturatosphaeria is distinct from the Lophiostomataceae clade. The aim of this study is to resolve the phylogenetic and morphological boundaries of Lophiostoma, Misturatosphaeria and several other allied genera in the family Lophiostomataceae sensu lato and to provide a backbone tree for this group of taxa.

Materials and methods Sample collection, specimen examination and isolation Type or representative specimens were loaned from Hirosaki University (HHUF), Research Institute of Resource Insects (IFRD) and the Swedish Museum of Natural History (S). Fresh collections were also made in Germany, Italy and Thailand between October 2012 and May 2013. Ascomata were rehydrated in 5 % KOH prior to examination and sectioning. Specimens were examined with a Motic SMZ 168 stereo microscope and fine forceps were used to remove ascomata, which were mounted in water. Sections of ascomata and conidiomata were made with a razor blade. Photographs were made from material mounted in water or lactophenol with cotton blue reagent using a Nikon ECLIPSE 80i light microscope with a Cannon 450D digital camera. Indian ink was added to water mounts to detect the presence of mucilaginous sheaths or ascospore appendages. Measurements were made with Tarosoft (R) Image Frame Work. Freshly collected samples were grown on potato dextrose agar (PDA). Isolates were derived via single spore isolation following the method of Chomnunti et al. (2014). Germinating spores were transferred to PDA and incubated at 25 °C in the dark. The cultural characteristics such as mycelium colour, shape, texture and growth rate were determined. Type and voucher specimens are deposited in the herbarium of Mae Fah Luang University (MFLU), Chiang Rai, Thailand, Hirosaki University (HHUF), Japan and Cryptogams, Kunming Institute of Botany (HKAS), China. The living cultures are deposited at the Mae Fah Luang University Culture Collection (MFLUCC) with duplicates in International Collection of Microorganisms from Plants (ICMP) Landcare Research, New Zealand, BIOTEC Culture Collection (BCC), Bangkok, Thailand and Kunming Institute of Botany Culture Collection (KUMCC). Faces of Fungi numbers are provided as in Jayasiri et al. (2015) and Index Fungorum numbers as in Index Fungorum (2015). DNA extraction, PCR amplification and sequencing Fungal isolates were grown on potato-dextrose agar (PDA) for 7–21 days at 25 °C and total genomic DNA was extracted from 50 to 100 mg of axenic mycelium scraped from the edges of the growing culture (Wu et al. 2001). Mycelium was ground to a fine powder with sterilized quartz sand (Sio2), 1 mg polyvinylpyrrolidone (PVP) and 200 μl of 2 % CTAB buffer using a sterilized


glass or plastic pestles in 1.5 ml eppendorf tubes. 400–500 μl of CTAB was added and incubated in 65 °C for about 60 min, while inverting the tubes several times and then, the tubes were centrifuged at 12,000 rpm for 10 min after cooling to room temperature. The supernatant was transferred in to sterilized microcentrifuge tubes and the equal volumes of Phenol: Chloroform: Iso Amyl Alcohol (25:24:1) were added. This was followed by mixing the solution by inversion and centrifuging the tubes at 10,000 rpm for 10 min (Chen et al. 2007). The supernatant resulted by the previous step was transferred to sterilized microcentrifuge tubes and the equal volumes of Chloroform: Iso Amyl Alcohol (24:1) solution were added. The solution was mixed by inversion and the tubes were centrifuged at 10, 000 rpm for 10 min. Upper aqueous phases (contains the DNA) were transferred to clean microfuge tubes and equal volumes of ice cold isopropanol were added to them. The microtubes were inverted gently to ensure mixing was complete and left for 1 h at −20 °C. Following the precipitation the tubes were spun at 12, 000 rpm for 10 min. The supernatant was removed carefully and the pellets were washed twice with 1 ml ice cold 75 % ethanol, followed by centrifuging at 12,000 rpm for 10 min. The Supernatant was removed and pellet dried in air for 15 min. The DNA was re-suspended in 50–100 μl of DNase free water. Polymerase chain reactions (PCR) were carried out using three partial gene regions in this study; LROR and LR5 (Vilgalys and Hester, 1990) for the nuclear ribosomal large subunit (LSU), ITS4 and ITS5 (White et al. 1990) for the internal transcribed spacer (ITS), EF1-983 F and EF1-2218R (Carbone and Kohn 1999) for translation elongation factor 1alpha (EF1-α) and NS1 and NS4 (White et al. 1990) the nuclear ribosomal small sub unit (SSU). The amplifications were performed in 25 μL of PCR mixtures containing 9.5 μL ddH2O, 12.5 μL 2×PCR Master Mix (TIANGEN Co., China), 1 μL of DNA template, 1 μL of each primer (10 μM). Conditions of amplification for all regions were consisted an initial denaturation step of 5 min at 94 °C and final extension step of 10 minutes at 72 °C. For the SSU and LSU amplification, the 37 cycles consisted of denaturation at 94 °C for 1 minute, annealing at 54 °C for 50 seconds and elongation at 72 °C for 1 minute; for the ITS amplification the 34 cycles consisted of denaturation at 94 °C for 30 seconds, annealing at 55 °C for 30 seconds and elongation at 72 °C for 1 minute; for the EF-1α amplification, the 40 cycles consisted of denaturation at 95 °C for 30 seconds, annealing at 58 °C for 50 seconds and elongation at 72 °C for 1 minute. The PCR products were viewed on 1 % agarose electrophoresis gels stained with Ethidium bromide. Purification and sequencing of PCR products were carried out at Invitrogen Biotechnology Co., Ltd. Shanghai, China. Sequence alignment and phylogenetic analyses LSU, SSU, ITS and EF-1α sequence data generated in this study were subjected to standard BLAST searches of GenBank for

Fungal Diversity (2015) 74:199–266 Table 2

203

GenBank and culture collection accession numbers of species treated in the phylogenetic study. Newly generated sequences are shown in bold

Old species name

New species name

Culture collection*

GenBank accession numbers LSU

Amorosia littoralis

EF1-α –

SSU

ITS

NN 6654

AM292055

Angustimassarina acerina Angustimassarina populi

MFLUCC 14-0505 MFLUCC 13-0034

KP888637 KR075168 KP899123 KP888642 KR075164 KP899128

KP899132 KP899137

Angustimassarina quercicola

MFLUCC 14-0506 CBS 128781

KP888638 KR075169 KP899124 JQ044450 – –

KP899133 JQ044431

Curreya austroafricana

Coelodictyosporium muriforme MFLUCC 13-0351 Neocurreya austroafricana CBS 122674

KP888641 KR075163 KP899127 EU552116 – EU552116

KP899136 EU552116

CBS 119330

EU552115

–

EU552115

EU552115

Curreya grandicipis

Neocurreya grandicipis

CBS 114272 CBS 111702

JN712520 JN712521

–

JN712456 JN712457

JN712456 JN712457

Curreya proteae Decaisnella formosa

Neocurreya proteae

CBS 122675 BCC 25616

EU552117 – EU552117 GQ925846 GU479851 GQ925833

EU552117 –

BCC 25617

GQ925847 GU479850 GQ925834

–

CPC 22679

KF777213

–

KF777160

CBS 610.74 CBS 160.30 JK 5603 K JK 5678I MFLUCC 15-0039

DQ384105 GU371821 DQ384067 JQ044447 – – GU479785 – GU479751 GU301813 GU479852 GU296149 KT305993 – KT305995

GQ203757 JQ044428 – – KT305997

MFLUCC 13-0442 MFLUCC 14-0993 BBH 22481 MFLUCC 11-0377

KP888639 KR075161 KP899125 KP888640 KR075162 KP899126 GU479786 – GU479752 KP888646 – –

KP899134 KP899135 – KP899140

Lignosphaeria thailandica MFLUCC 11-0376 Lophiopoacea paramacrostoma MFLUCC 11-0463 MFLUCC 15-0701

KP888645 – – KP888636 – KP899122 KT333436 KT427535 KT333437

KP899139 – KT333435

Camarographium carpini

Dimorphiopsis brachystegiae Eremodothis angulata Exosporium stylobatum Floricola striata Floricola viticola Guttulispora crataegi Halotthia posidoniae Lignosphaeria fusispora

Lophiohelichrysum helichrysi Lophiostoma alpigenum Lophiostoma arundinis

AM292056 AM292047

–

Sigarispora caudata Sigarispora caulium

GKM 1091b JCM 13550 JCM 13551/MAFF 239449 MAFF 239453 MAFF 239450

GU385193 AB618998 AB618999 AB619000 AB619001

– AB618679 AB618680 AB618681 AB618682

– JN942964 JN942965 LC001723 LC001724

Lophiostoma caulium Bvar. d^

Lophiostoma multiseptatum

JCM 17669 MFLUCC 15-0176 JCM 17668

AB619002 LC001741 AB618683 KT328493 – KT328494 AB619003 LC001742 AB618684

LC001725 – LC001726

Lophiostoma caulium Bvar. f^

Biappendiculispora japonica

MAFF 239451 MAFF 239452

AB619004 LC001743 AB618685 AB619005 LC001744 AB618686

LC001727 LC001728

JCM 17671 JCM 17670 KT 521 JCM 13125/ MAFF 239635 IFRD 2014 CBS 629.86 JCM 17672 MAFF 239458

AB619007 AB619006 JN941380 JN941379 FJ795437 DQ678069 AB619008 AB619009

LC001730 LC001729 JN942963 JN942962 – – LC001731 LC001732

Lophiostoma caudatum Lophiostoma caulium Bvar. a^

Sigarispora arundinis

Lophiostoma compressum

Platystomum actinidiae

Lophiostoma crenatum Lophiostoma fuckelii

Vaginatispora fuckelii

GU327758 LC001737 LC001738 LC001739 LC001740

LC001746 LC001745 LC001747 LC001748 – DQ677912 LC001749 LC001750

AB618688 AB618687 JN941375 JN941376 FJ795480 DQ678017 AB618689 AB618690

204


Table 2 (continued) Old species name

New species name

Culture collection*


EF1-α

SSU

ITS

Lophiostoma heterosporum

CBS 644.86

AY016369 DQ497609 AY016354

Lophiostoma macrostomoides Lophiostoma macrostomum

CBS 123097

FJ795439

JCM 13544

AB619010 LC001751 AB618691

JN942961

JCM 13546/ MAFF 239447 AB433274 LC001753 AB521732

AB433276

JCM 13545 MFLUCC 13-0451

AB433273 LC001752 AB521731 KR025862 – –

AB433275 KR025858

GKM1233

GU385184 GU327760

Lophiostoma pseudodictyosporium Lophiostoma quadrinucleatum Lophiostoma quadrisporum

Coelodictyosporium pseudodictyosporium

GU456277 FJ795482

–

GQ203795 –

–

Paucispora quadrispora

KH 448

LC001722 LC001754 LC001720

LC001733

Lophiostoma ravennicum

Sigarispora ravennica

MAFF 239455/KT 843 MFLUCC 14-0005

AB619011 LC001755 AB618692 KP698414 – KP698415

LC001734 KP698413

Lophiostoma sagittiforme Lophiostoma semiliberum

Capulatispora sagittiformis

JCM 15100 JCM 13548

AB369267 LC001756 AB618693 AB619012 LC001757 AB618694

AB369268 JN942966

JCM 13547 AB619013 LC001758 AB618695 JCM 13549/MAFF 239448 AB619014 LC001759 AB618696

JN942967 JN942970

Lophiostoma triseptatum

SMH 2591 SMH 5287

GU385183 GU385187

– –

– –

– –

Lophiostoma vaginatispora Lophiostoma versicolor Lophiostoma viridarium

Vaginatispora aquatica Paucispora versicolor

MFLUCC 11-0083 MAFF 244508 IFRDCC 2090

KJ591576 – KJ591575 AB918732 LC001760 LC001721 FJ795443 – FJ795486

KJ591577 AB918731

Lophiostoma vitigenum

Pseudolophiostoma vitigenum

Lophiostoma winteri

Lophiopoacea winteri

JCM 13534/MAFF 239459 JCM 17676 JCM 17648/KT 740 MAFF 239454 MFLUCC 12-0088

AB619015 AB619016 AB619017 AB619018 KF531927

LC001735 LC001736 JN942969 JN942968 –

CBS 154.93 BCC 28866 BCC 28867 CBS 124080

FJ795448 – FJ795491 GU371824 GU371817 GU371832 GU371825 GU371818 GU371833 GU456323 GU456265 GU456302

– –

ATCC 42522 GKM 1238

U43479 – U43461 GU385173 GU327761 –

– –

GKM 1280 GKM 1210 SMH 5151

GU385174 GU327762 GU385212 GU327763 GU385211 –

– – –

– – –

GKM L100Na

GU385189 GU327766

–

–

GU385194 GU385188 JN851819 GU385165 GU385182 GU385195 GU385166 GU385207

Macrodiplodiopsis desmazieri Magnibotryascoma sp. Massarina corticola Mauritiana rhizophorae Melanomma pulvis-pyrius Melanomma radicans Misturatosphaeria aurantiinotata

Misturatosphaeria radicans

Misturatosphaeria claviformis Neocurreya claviformis Misturatosphaeria cruciformis Pseudomisturatosphaeria cruciformis Misturatosphaeria kenyensis Pseudoaurantiascoma kenyense

Misturatosphaeria mariae Misturatosphaeria minima

Asymmetrispora mariae Aurantiascoma minimum

Misturatosphaeria tennesseensis

Asymmetrispora tennesseensis

GKM1195 GKM234N CBS 124079 GKM 169 N ANM 60 ANM933 SMH2448 ANM 911

Misturatosphaeria radicans

SMH 4330

LC001761 LC001762 LC001763 LC001764 –

AB618697 AB618698 AB618699 AB618700 KF531928

–

GU327767 GU327765 KR075166 GU327768 –

–

–

– – –

– – –

GU327769

–

–

GU385167 GU327770

–

–


205

Table 2 (continued) Old species name

New species name

Culture collection*


Misturatosphaeria uniseptata Misturatosphaeria uniseriata Misturatosphaeria sp.

Platystomum scabridisporum

Magnibotryascoma uniseriatum ANM 909

EF1-α

SSU

ITS

GU385206

–

–

–

–

–

–

SMH 3747

GU385196

Platystomum compressum

MFLUCC 13-0343

KP888643 KR075165 KP899129

–

Platystomum crataegi Platystomum rosae


KT026109 KT026121 KT026113 KT026111 – KT026115

KT026117 KT026119

Platystomum salicicola

MFLUCC 15-0632

KT026110

–

KT026118

Pseudoplatystomum scabridisporum

BCC 22835

GQ925844 GU479857 GQ925831

KT026114

–

BCC 22836

GQ925845 GU479856 GQ925832

–

Preussia funiculata

CBS 659.74

GU301864 GU349032 GU296187

–

Preussia lignicola Preussia terricola

CBS 264.69 DAOM 230091

GU301872 GU349027 GU296197 AY544686 DQ471063 AY544726

– AY544726


KT026112 – KT026116 KP888647 KR075167 KP899131

KT026120 KP899141

MFLUCC 10-0126

KP888644 KR075170 KP899130

KP899138

CBS 524.50 GKM 1033 GKM 1159 KTC 1124

DQ678056 DQ677897 DQ678003 GU385190 GU327776 – GU385185 GU327778 – GU205227 – GU205256

– – – –

SC-12 CBS 454.72 CBS 508.75

JX985750 – JX985749 JN662930 AY004343 DQ497610 AY016355 DQ491519 DQ468050 – WDU42488 GQ203798

CBS 379.55

GU301880 GU349021 GU296208

Sigarispora coronillae Ramusculicola thailandica Sporormiella minima Thyridaria macrostomoides Trematosphaeria biappendiculata Trematosphaeria terricola Westerdykella cylindrica Westerdykella dispersa Westerdykella ornata

Neotrematosphaeria biappendiculata Alpestrisphaeria terricola

NR103587

*ANM: A.N. Miller, ATCC: American Type Culture Collection, Virginia, USA, BCC: BIOTEC Culture Collection, Bangkok, Thailand, BBH: BIOTEC Bangkok Herbarium, Thailand; CBS: Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, CPC: Working collection of Pedro Crous housed at CBS, DAOM: Plant Research Institute, Department of Agriculture (Mycology), Ottawa, Canada, GKM: G.K. Mugambi, IFRDCC: Culture Collection, International Fungal Research and Development Centre, Chinese Academy of Forestry, Kunming, China, JCM: the Japan Collection of Microorganisms, Japan, JK: J. Kohlmeyer, KT: K. Tanaka, KH: K. Hirayama, MAFF: Ministry of Agriculture, Forestry and Fisheries, Japan, MFLUCC: Mae Fah Luang University Culture Collection, ChiangRai, Thailand, NN: NovoNordisk culture collection (now Novozymes, Bagsvaerd, Denmark), SC: Department of Plant Pathology, Sichuan Agricultural University, SMH: S.M. Huhndorf

rough identification. Sequences of the available ex-type cultures and other taxa were obtained from GenBank (Table 2). Several other taxa from closely related families (Halotthiaceae and Sporormiaceae) were also included in the analyses. The consensus sequences for each gene were initially aligned by ClustalX v. 1.83 and in Bioedit (Thompson et al. 1997). LSU, SSU, ITS and EF1-α datasets were first analyzed separately and single LSU, SSU, ITS and EF1-α gene trees reflect the same underlying phylogeny (no significant differences). Therefore, the individual datasets were concatenated into a combined dataset. A Maximum likelihood analysis was performed at CIPRES using RAxML v.7.2.8 as part of the BRAxML-HPC2 on TG^ tool (Stamatakis et al. 2008). The general time reversible model (GTR) using proportion of invariable sites was applied with a

discrete gamma distribution and four rate classes. The best scoring tree was selected with a final likelihood value of −25, 138.092571. The sequences of novel species and other newly generated sequences are deposited in GenBank. The model of evolution was performed using MrModeltest 2.2 (Nylander 2004). Posterior probabilities (PP) (Rannala and Yang 1996; Zhaxybayeva and Gogarten 2002) were determined by Markov Chain Monte Carlo sampling (MCMC) in MrBayes v. 3.0b4 (Huelsenbeck and Ronquist 2001). Six simultaneous Markov chains were run for 10,000,000 generations and trees were sampled every 100th generation. The first 100,000 trees, representing the burn-in phase of the analyses, were discarded, while the remaining trees (80,000) were used for calculating posterior probabilities in the majority rule

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consensus tree. Bayesian Posterior Probabilities (BYPP) equal or greater than 0.90 are given below or above each node (Fig. 1). Phylogenetic trees were drawn using Treeview (Page 1996), FigTree v. 1.4 (Rambaut and Drummond 2008) and MEGA5 (Tamura et al. 2011).

Results Phylogenetic analyses One hundred and nine strains are included in the combined LSU, SSU, ITS and EF-1α gene analyses with Melanomma pulvis-pyrius (CBS 124080) as the outgroup taxon. Tree topology of the Bayesian analysis was compatible with the ML tree and was not significantly different (data not shown). The best scoring RAxML tree with a final likelihood value of −25, 138.092571 is shown in Fig. 1. The main Lophiostoma clades (A and B) comprise 56 strains, which represents a relatively small number of taxa. Many of the strains used in this tree come from previous studies for which there is no morphological data and they must be regarded as putatively named strains. Lophiostoma species formed two main clades (A and B) with 100 % ML and 1.00 PP values. Clade A separated into 14 subclades (A1–A14) and the uppermost clade (A1), comprising several putative strains of L. compressum (Pers.) Ces. & De Not., L. viridarium Cooke and L. triseptatum Peck. including our strain of Platystomum compressum and three new Platystomum species which represents the genus Platystomum. Lophiostoma sensu stricto (A2), comprises the putative Lophiostoma strains L. semiliberum (JCM 13547, 13548, 13549), L. crenatum (CBS 629.86) and L. multiseptatum (L. caulium Bvar. d^ - JCM 17668, MAFF 239452), along with the type strain of Lophiostoma, L. macrostomum. Subclade A3 comprised Lophiostoma pseudodictyosporium and our new strain MFLUCC 13-0351 for which the new genus Coelodictyosporium is introduced. S u b c l a d e A 4 , N e o t re m a t o s p h a e r i a , c o m p r i s e s Trematosphaeria biappendiculata Kaz. Tanaka et al. Subclade A5, the new genus Sigarispora consists of Lophiostoma arundinis, L. caudatum, L. caulium Bvar. a^, L. ravennicum and a new species MFLUCC 14-0941. Lophiostoma (Lophiopoacea) winteri and a new species MFLUCC 110463 grouped in the subclade A6 for which a new genus Lophiopoacea is introduced. The newly introduced genus Lophiohelichrysum is represented by subclade A7. Subclade A8 containing two new strains MFLUCC 13-0442 and 140585 are incorporated in the new genus Guttulispora. The new genus Capulatispora (A9) is represented by Lophiostoma sagittiforme Kaz. Tanaka & Hosoya, while two putatively named strains of Lophiostoma (Trematosphaeria) heterosporum (De Not.) G. Winter and L. macrostomoides De Not. clustered in the unnamed subclade A10. Platystomum


scabridisporum Abdel-Wahab & E.B.G. Jones (BCC 22835/ 36) grouped in subclade A11 which represents the new genus Pseudoplatystomum. Subclade A12 comprised Lophiostoma caulium Bvar. f^ which is proposed a new genus Biappendiculispora and the ex-type strain of Lophiostoma vitigenum (Kaz. Tanaka & Y. Harada) K. Hiray. & Kaz. Tanaka (JCM 13534) clustered in subclade A13 which is named as the new genus Pseudolophiostoma. The final subclade (A14) in clade A represents Trematosphaeria terricola G.S. Gong which belongs to the new genus Alpestrisphaeria. Clade B contains six Lophiostoma species and the recently introduced Dimorphiopsis brachystegiae Crous (CPC 22679), the type of Dimorphiopsis. These species were separated into four subclades (B1-B4). Subclade B1 incorporates two putative strains of Lophiostoma fuckelii Sacc. (MAFF 239458 and JCM 17672), Vaginatispora aquatica K.D. Hyde and Lophiostoma alpigenum Fuckel which belong in the genus Vaginatispora K.D. Hyde. Paucispora is a new genus represented by subclade B2 which consists of Lophiostoma quadrisporum K. Hiray. & Kaz. Tanaka and L. versicolor K. Hiray. & Kaz. Tanaka. Dimorphiopsis brachystegiae Crous clusters in subclade B3. Subclade B4 remains as an unnamed clade which comprises Lophiostoma quadrinucleatum P. Karst. All species of Misturatosphaeria, together with new species (MFLUCC10–0126 and MFLUCC 13-0284), Floricola striata Kohlm. & Volkm.-Kohlm. (JK 5603K and JK 5678I), Floricola viticola Phukhamsakda et al. (MFLUCC 15-0039), Curreya austroafricana Marinc. et al., C. grandicipis (Joanne E. Taylor & Crous) Joanne E. Taylor & Crous and C. proteae Marinc. et al. form a well-supported clade (C) in the phylogenetic analyses. Misturatosphaeria species separated into nine subclades and six new genera are introduced for them. In this paper we introduce a new family, Floricolaceae, to accommodate taxa in this clade. Clade C1 represents the genus Floricola, while subclade C2 containing Misturatosphaeria mariae Zhang et al. and M. tennesseensis Mugambi et al. are incorporated in the new genus Asymmetrispora. The subclade C3 belongs to the new genus Pseudomisturatosphaeria which is introduced for Misturatosphaeria cruciformis Mugambi & Huhndorf. The new genus Pseudoaurantiascoma (C4) is represented by M. kenyensis Mugambi & Huhndorf. (Pseudoaurantiascoma kenyense), while the genus Misturatosphaeria (C5) consists of M. aurantiinotata Mugambi & Huhndorf, the type species of Misturatosphaeria, M. uniseptata (radicans) Mugambi et al. and an undescribed Fig. 1 RAxML tree based on a combined LSU, SSU, ITS and TEF1α sequence dataset. Bootstrap support values for maximum likelihood (ML) equal or greater than 50 % and Bayesian posterior probabilities (BYPP) equal or greater than 0.90 are given above and below the nodes respectively. Ex-type culture numbers are in bold. The tree is rooted to Melanomma pulvis-pyrius (CBS 124080)


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Misturatosphaeria species (SMH 3747). Two species (MFLUCC10-0126 and MFLUCC 13-0284) grouped in subclade C6 which represents the new genus Ramusculicola. Misturatosphaeria claviformis Mugambi & Huhndorf., Curreya austroafricana, C. grandicipis and C. proteae clustered in subclade C7 for which the new genus Neocurreya is introduced. The subclade C8 is incorporated in the new genus Magnibotryascoma which is introduced for M. uniseriata Mugambi et al. and subclade C9 belongs to the new genus Aurantiascoma which comprises M. minima Mugambi et al. Clades D and E show close phylogenetic affinities with Lophiostomataceae and Floricolaceae. Clade D includes putative strains of Decaisnella formosa (BCC 25616 and BCC 25617), Thyridaria macrostomoides (GKM 1033 and GKM 1159) and two new strains (MFLUCC 11-0377 and MFLUCC 11-0376) from this study, which represent a new genus, Lignosphaeria. Clade E comprises two major subclades (E1 and E2) with 93 % ML/1.00 B.P. support, with one subclade (E2) representing the monotypic genus Amorosia. Another subclade (E1) contains three species (MFLUCC 14-0505, MFLUCC 13-0034 and MFLUCC 14-0506) from this study, and putatively named strains of Exosporium stylobatum (CBS 160.30) and Massarina corticola (CBS 154.93). We introduce a new family, Amorosiaceae, based on phylogeny and unique morphological characters, for the taxa in clade E. A new genus, Angustimassarina, is introduced to accommodate three new species from new collections. Camarographium carpini grouped as a separate clade in between clade E (Amorosiaceae) and Sporormiaceae with low bootstrap support. Halotthiaceae and Sporormiaceae appeared as basal clades and these families can be considered as the closest relatives of Lophiostomataceae, Floricolaceae, and Amorosiaceae. Lophiostomataceae Sacc. [as ‘Lophiostomaceae’], Syll. Fung. (Abellini) 2: 672 (1883); Facesoffungi number: FoF00796 Saprobic on twigs or bark of various woody plants and herbaceous plants in terrestrial and aquatic environments. Sexual morph: Ascomata solitary or scattered to gregarious, immersed to erumpent, coriaceous to carbonaceous, dark brown to black, globose to subglobose, uniloculate, ostiolate. Ostiole, rounded or slit-like, with a small to large, compressed, periphysate or not, crest or slot-like apex, which may be poorly developed or lacking, variable in shape and composed of pseudoparenchymatous cells. Peridium thick at the sides, broad at the apex and thinner at the base, one to several layers, composed of small to medium, dark brown or lightly pigmented to hyaline, thinwalled cells of textura prismatica or textura angularis, fusing and indistinguishable from the host tissues. Hamathecium comprising numerous, septate or aseptate, anastomosing and branched or unbranched, cellular or filamentous pseudoparaphyses, situated between and above


the asci, embedded in a gelatinous matrix. Asci 2–8spored, bitunicate, fissitunicate, cylindrical to clavate, pedicellate, rounded at the apex, with an ocular chamber. Ascospores uni to bi-seriate, partially overlapping, hyaline or yellowish to brown, sometimes with hyaline end cells, fusiform or ellipsoid to fusiform, with acute or rounded ends, 1 to multi-septate or muriform, and constricted at the central septum, often guttulate, smooth-walled or verruculose, sometimes with terminal appendages, with or lacking a mucilaginous sheath. Asexual morph: coelomycetous. Conidiomata dark brown to black, immersed to erumpent to superficial, solitary or scattered, sometimes in groups, unilocular, globose to subglobose covered with brownish, septate, hyphae, ostiolate. Ostiole central, circular. Conidiomtal wall thick at the sides, slightly thinner at the base, composed of dark brown to lightly pigmented, compressed cells of textura angularis. Conidiophores cylindrical, septate, branched, hyaline, and formed all around the locular cavity. Conidiogenous cells phialidic, holoblastic, integrated, hyaline or brown, smooth, cylindrical or ampulliform. Conidia cylindrical to ellipsoid, aseptate to multi-septate or 1-distoseptate, sometimes consisting of 6–8 rows of cells, each row comprising 5–7 cells, hyaline or light brown to brown, smooth-walled (asexual morph description follows Tanaka and Harada 2003a; Crous et al. 2013; Hyde et al. 2013, Liu et al. 2015). Notes: In this study we accept 16 genera in Lophiostomataceae, including eleven new genera. We propose a new family Floricolaceae which contains Floricola, Misturatosphaeria and seven other new genera based on our phylogenetic analyses and morphology. The families Lophiostomataceae and Floricolaceae formed wellsupported clades in our phylogenetic analysis (Fig. 1), while the Lophiostoma species separate in two distinct clades (A and B). Clade A comprises 13 genera, including Lophiostoma sensu stricto. Clade B separates distinctly from clade A and contains six Lophiostoma species and the ex-type strain of Dimorphiopsis brachystegiae (CPC 22679). Dimorphiopsis is a recently introduced genus with D. brachystegiae as its type. Crous et al. (2013) accommodate this genus in Lophiostomataceae using LSU and ITS sequence data. It is difficult to distinguish if this species is a coelomycete or hyphomycete due to its unusual morphology (Crous et al. 2013). The Dimorphiopsis type of asexual morph is not evident in other species in clade B. The other species in clade B share some similar morphological characters with Lophiostoma sensu stricto, such as immersed to erumpent, carbonaceous ascomata with slit-like ostioles, cylindrical to clavate asci and 1 to multi-septate ascospores (Tanaka and Harada 2003a, Zhang et al. 2014). They, however, differ in peridium, asci and ascospores characters and phylogenies. Therefore, we accept Dimorphiopsis, Paucispora and Vaginatispora as


distinct genera in this clade, based on morphology and phylogenetic distance from Lophiostoma sensu stricto. Key to genera of Lophiostomataceae 1. Asexual morph difficult to distinguish between a coelomycete or a hyphomycete, with pale to dark brown, 1septate conidia with obtuse ends and flattened basal scar, asexual morph unknown . . . . . . . . . . . . . . . . . . . Dimorphiopsis 1. Asexual m orph unknown, or not as above, coelomycetous when asexual morph known . . . . . . . . . . . . 2 2. Ascomata covered with sparse, brown, septate hyphae . . . . . . . . . . . . . . . . . . . . . . . . . . . . Alpestrisphaeria 2. Ascomata without hyphal covering . . . . . . . . . . . . . . 3 3. Asci 2-4-spored . . . . . . . . . . . . . . . . . . . . Paucispora 3. Asci 8-spored. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4. Ascomata conical. . . . . . . . . . . . . . . . . .Capulatispora 4. Ascomata usually globose to subglobose . . . . . . . . . . 5 5. Ascomata semi-immersed to erumpent . . . . . . . . . . . . 6 5. Ascomata immersed, only papilla erumpent through host surface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 6. Ascospores lacking a mucilaginous sheath and terminal appendages. . . . . . . . . . . . . . . . . . . . . . Lophiohelichrysum 6. Ascospores with a mucilaginous sheath or terminal appendages. . . . . . . . . . . . . . . . . . . . . . . . . . . Vaginatispora 7. Ostiole broadly papillate . . . . . . . . . . . . . . . . . . . . . . 8 7. Ostiole usually not as above. . . . . . . . . . . . . . . . . . . 11 8. Peridium composed of cells of textura angularis, ascospores muriform with both transverse and vertical septa. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 8. Peridium composed of cells of textura prismatica, ascospores 1-multiseptate with only transverse septa . . . . . . . . . 8 9. Ascospores brown, with verrucose or rough-walls, lacking a mucilaginous sheath or terminal appendages. . . . . . . . . . . . . . . . . . . . . . Pseudoplatystomum 9. Ascospores yellowish to brown, smooth-walled, with a mucilaginous sheath and terminal appendages. . . . . . . . . . . . . . . . . . . . . . Coelodictyosporium 10. Ascospores pale yellowish to olivaceous brown, except at the pale end cells, 5(–7)-septate, with appendages at both ends . . . . . . . . . . . . . . . . . . . . . . . . . . . Neotrematosphaeria 10. Ascospores hyaline, 1-septate, with 2 eusepta and a narrow mucilaginous sheath, lacking terminal appendages. . . . . . . . . . . . . . . . . . . . . . Pseudolophiostoma 11. Ascospores muriform or multi-septate, with both transverse and vertical septa. . . . . . . . . . . . . . . . . . . . . . . . . . . 12 11. Ascospores 1-multi-septate, with only transverse septa. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 12. Ascospores 5–6 transversely septate, rarely with vertical septa, with or without a mucilaginous sheath or terminal appendages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . Sigarispora 12. Ascospores usually muriform, lacking a mucilaginous sheath and terminal appendages . . . . . . . . . . . .Platystomum

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13. Ascospores mostly hyaline, rarely l ightly pigmented . . . . . . . . . . . . . . . Lophiopoacea, Lophiostoma 13. Ascospores pigmented, pale yellowish to brown. . . 14 14. Ascospores ellipsoid to fusiform, straight, brown, 3septate, lacking terminal appendages . . . . . . . . Guttulispora 14. Ascospores fusiform, curved, pale yellowish to brown, 7–9-septate, with terminal appendages . . . . . . . . . . . . . . . . . . . Biappendiculispora Type : Lophiostoma Ces. & De Not., Comm. Soc. crittog. Ital. 1(4): 219 (1863) Facesoffungi number: F0F00797 Saprobic on herbaceous and woody substrates in terrestrial and aquatic habitats. Sexual morph: Ascomata scattered to gregarious, immersed to semi-immersed, papilla erumpent through host surface, coriaceous to carbonaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole slit-like, variable in shape, with crest-like apex, usually opening apically with a pore, plugged by gelatinous tissue and occasionally lighter coloured. Peridium wider at the apex and thinner at the base, composed of a single stratum, comprising several layers of lightly pigmented to dark brown, thin-walled cells of textura angularis to textura prismatica, cells towards the inside lighter, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2 μm wide, septate, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, short to long pedicellate, rounded at the apex with an ocular chamber. Ascospores uniseriate or partially bi-seriate, hyaline to yellowish brown, fusiform with narrow, acute ends, 1 to multi-septate, sometimes with 3–5-eusepta, constricted at the central septum, with or without terminal appendages, lacking a mucilaginous sheath. Asexual morph: see under Lophiostoma semiliberum. Notes: Lophiostoma was established by Cesati and De Notaris (1863) with L. macrostomum as its type (Tanaka and Harada 2003a; Hyde et al. 2013). Lophiostoma is morphologically a well-studied genus (Chesters and Bell 1970; Holm and Holm 1988; Barr 1990; Yuan and Zhao 1994; Tanaka and Harada 2003a; Zhang et al. 2009). Chesters and Bell (1970) treated Lophiotrema as a synonym of Lophiostoma, but later Holm and Holm (1988) distinguished Lophiotrema from Lophiostoma based on the differences in the type of peridium and asci, while Hirayama and Tanaka (2011) separated them on the basis of both morphological characters and molecular analyses. The genus Platystomum shares many characters with Lophiostoma (Tanaka and Harada 2003a) and Holm and Holm (1988) treated Platystomum as a synonym of Lophiostoma. Barr (1990) and Tanaka and Harada (2003a) distinguished Lophiostoma and Platystomum, while Mugambi and Huhndorf (2009) revealed that the putatively named strain (GKM1048) of the type of Platystomum, P. compressum clustered outside of Lophiostomataceae in their phylogenetic analysis. However, in our phylogenetic

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analysis our collection of P. compressum grouped in clade A and therefore, we accept Platystomum in Lophiostomataceae. Combined gene analysis of LSU and SSU sequence data showed Lophiostoma to be polyphyletic, grouping into two well-supported monophyletic clades (Suetrong et al. 2009; Zhang et al. 2009). Kirk et al. (2008) estimated that there are 83 species in Lophiostoma and 446 species epithets are listed in Index Fungorum (2015). Species which grouped in the subclade A2 in our phylogenetic tree are considered as Lophiostoma sensu stricto. Lophiostoma caulium and allied taxa have been considered to be a species complex by different authors (Chesters and Bell 1970; Holm and Holm 1988), the latter who described varieties of this species. Holm and Holm (1988) introduced five varieties and named them as L. caulium var. a to e, mainly on the basis of ascospore size and septation, but none of them are formally published. Tanaka and Harada (2003a) analyzed and illustrated three varieties (a, d, f) and their morphology are different in many aspects. Hirayama and Tanaka (2011) included these three varieties in their phylogenetic analysis and they formed separate sub-clades in clade A. In our study, L. caulium var. a, d and f clustered in three separate sub-clades (A2, A5 and A12) in clade A and these are clearly shown to be three distinct species in both studies. Therefore we accommodate L. caulium Bvar. a^ (L. caulium (Fr.) Ces. & De Not.,) in Sigarispora which grouped in the Sigarispora clade (A5) based on morphology and phylogeny. L. caulium Bvar. d^ clustered in the Lophiostoma clade (A2) and can be considered as a new Lophiostoma species. A new genus is proposed for the L. caulium Bvar. f^ which grouped in the clade A12. Futher studies focusing on epitypification and molecular analyses are needed for the other verities in order to establish natural classification. Lophiostomataceae is a morphologically highly diverse family with many genera and species. Therefore, we establish Lophiostoma sensu stricto (A2) with the type species, while separating other species into 13 genera based on morphology and phylogeny. Lophiostoma sensu stricto is characterized by semi-immersed to densely erumpent, coriaceous to carbonaceous ascomata with crest and slit-like ostioles and hyaline to lightly pigmented, 1 to multi-septate ascospores with terminal appendages. Type species: Lophiostoma macrostomum (Tode, Fungi mecklenberg. sel. (Lüneburg) 2: 12 (1791) Ces. & De Not., Comm. Soc. Critt. Ital. 1(4): 219, 1863 Facesoffungi number: FoF00797; Figs. 2 and 3 Basionym: Sphaeria macrostoma Tode: Fr., Syst Mycol 2: 469, 1823 Saprobic on herbaceous and woody substrates in terrestrial and aquatic habitats. Sexual morph: Ascomata 450–555μm high×514–580μm diam. (x = 500×540μm, n=10), scattered to gregarious, semi-immersed, coriaceous to carbonaceous, dark brown to black, globose to subglobose. Ostiole slit-like, variable in shape, up to 650μm long and 530μm wide, central,


papillate, with a crest-like apex, usually opening apically with a pore, plugged by gelatinous tissue, made up of hyaline, pseudoparenchymatous cells and occasionally lighter coloured. Peridium 25–45μm wide (x = 37μm, n=12), wider at the apex and thinner at the base, composed of a single stratum, with several layers of hyaline to brown cells of textura angularis to textura prismatica, cells towards the inside lighter and somewhat flattened, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2 μm wide, septate, cellular pseudoparaphyses, anastomosing and branched, between and above the asci, embedded in a gelatinous matrix. Asci 94.5– 130.5 × 10–13 μm (x = 115.5 × 11.2 μm, n = 10), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a short, bulbous pedicel, rounded at the apex, with an ocular chamber. Ascospores 27–38.5×4.3–6.9μm (x = 34.5×5.3μm, n=15), uniseriate to bi-seriate, ascospores hyaline becoming pale brown, fusiform, with narrow, acute ends, usually 1-septate with 3–5-eusepta, constricted at the central septum, cells above central septum swollen, guttulate, smooth-walled, with terminal appendages. Asexual morph: Undetermined. Material examined: FRANCE, Ariège, Rimont, Las Muros, on dead stems of Vitis vinifera (Vitaceae), 2 September 1996 (IFRD 2005, epitype); JAPAN, Sanpinai, Hirosaki, Aomori, on stems of unknown herbaceous plant, 5 August 2001, Kazuaki Tanaka KT 635 (HHUF 27290), living culture, JCM 13545; JAPAN, Oowasawa River, river bank, Kadoke, Hirosaki, Aomori, on stems of an unknown plant, 25 August 2001, KT 709 (HHUF 27293), living culture JCM 13546/ MAFF 239447; JAPAN, Sanpinai, Hirosaki, Aomori, On stems of an unknown herbaceous plant, 5 August 2001, KT 635 (HHUF 27290), living culture JCM 13545; JAPAN: Campus of Hirosaki University, Hirosaki, Aomori, On twigs of Morus bombycis (Moraceae), KT 508 (HHUF 27288), living culture JCM 13544. Lophiostoma semiliberum (Desm.) Ces. & De Not., Schem. di Classif. Sferiacei: 46 (1863) Basionym: Sphaeria semilibera Desm., Annls Sci. Nat., Bot., sér. 3 6: 78 (1846); Fig. 4 Saprobic on dead culms of Phragmites australis (Cav.) Trin. ex Steud. Sexual morph: Ascomata 319–366 μm high×330–370μm diam. (x = 345×356μm, n=10), scattered to gregarious, immersed to semi-immersed, coriaceous to carbonaceous, dark brown to black, globose to subglobose. Ostiole slit-like, central, with a small crest-like apex, with a pore-like opening, plugged by gelatinous tissue, made up of hyaline, pseudoparenchymatous cells. Peridium 56–60μm wide(x = 58μm, n=12), wider at the apex and thinner at the base, composed of a single stratum, with several layers of lightly pigmented to reddish brown cells of textura angularis, cells towards the inside lighter, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1.5–2 μm wide, septate, branched, cellular


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Fig. 2 Lophiostoma macrostomum (epitype). a, b. Appearance of ascomata on host surface c–e, j. Sections of ascomata; note small to large, flat, crest-like papilla f–i. Asci with ascospores k–n. Ascospores with terminal appendages o. Pseudoparaphyses Scale bars: b, c=200μm, d=100μm, e=50μm, f–o=10μm

pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 84–114×8–12μm (x = 96×10.2μm, n=10), 8-spored, bitunicate, fissitunicate, cylindrical, pedicellate, rounded at the apex, with an ocular chamber. Ascospores 24–28.5×3–4.2μm (x = 25.5×3.9μm, n=15), uniseriate or partially bi-seriate, hyaline, fusiform with narrow, acute ends, usually 1-septate, constricted at the septum, guttulate, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Conidiomata scattered or sometimes in groups, subglobose with a flattened base, covered with

brownish septate hyphae. Ostiole central, papillate to cylindrical. Conidiomatal wall wide at the side, slightly thinner at the base composed of hyaline, compressed cells of textura angularis. Conidiophores septate, branched, hyaline, and formed all around the locular cavity. Conidiogenous cells phialidic. Conidia, ellipsoid to obovoid, aseptate, hyaline, smooth (asexual morph description follows Tanaka and Harada 2003a). Material examined: JAPAN, Hirakawa river bank, Hiraka, Aomori, on culms of Phragmites australis (Poaceae), 05

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Fig. 3 Lophiostoma macrostomum (HHUF 27290). a, b. Specimens and herbarium label. c, d. Appearance of gregarious ascomata on host surface e, f. Sections of ascomata; note ostiole with a small to large, flat, crest-like papilla. g. Ostiole. h. Section through peridium (from HHUF 27293) i–l. Asci with ascospores. m. Pseudoparaphyses. n–p. Ascospores. Scale bars: c, d= 200μm, e, f=50μm, g, h=25μm, i-l=20μm, m-p=10μm

August 2001, K. Tanaka KT 652 (HHUF 27298), living culture JCM 13547; JAPAN, Oowasawa River, river bank, Kadoke, Hirosaki, Aomori, On culms of an unknown Gramineae, 4 August 2001, KT 622 (HHUF 27299), living culture JCM 13548; JAPAN, Oowasawa River, river bank, Simizumori, Hirosaki, Aomori, on culms of an unknown herbaceous plant, 28 October 2001, KT 828 (HHUF 27300), living culture JCM 13549/MAFF 239448. Lophiostoma multiseptatum Thambugala, Wanasinghe, Kaz. Tanaka, & K.D. Hyde, sp. nov. Index Fungorum number: IF551527; Facesoffungi number: FoF00798; Fig. 5 Etymology: Referring to its multi-septate ascospores.

Saprobic on dead herbaceous twigs in terrestrial habitats. Sexual morph: Ascomata 250–450μm high, 180–400μm diam, solitary, immersed, papilla erumpent through host surface, coriaceous to carbonaceous, black, subglobose, ostiolate. Ostiole slit-like, variable in shape, up to 80–220μm long and 30–80μm wide, central, papillate, with a crest-like apex and an irregular pore-like opening, plugged by a gelatinous tissue, made up of hyaline, pseudoparenchymatous cells and occasionally lighter coloured. Peridium 35– 55μm wide (x = 44.7μm, n=20), wider at the apex and thinner at the base, composed of a single stratum, with several layers of lightly pigmented to brown cells of textura prismatica, cells towards the inside lighter and somewhat broad, at the outside, darker, fusing and


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Fig. 4 Lophiostoma semiliberum (HHUF 27298). a. Herbarium material b, c. Appearance of ascomata on the host surface d. Section through ascoma e. Section through peridium f. Pseudoparaphyses g– h Immature ascus i. Mature bitunicate asci j–m. Ascospores. Scale bars: d =100μm, e=30μm, f=5μm, g-i=30μm, j–m=10μm

indistinguishable from the host tissues. Hamathecium comprising numerous, 2–3μm (n=30) wide, filamentous, septate, branched, pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 80–120×12–20μm (x = 101.3×14.6μm, n=40), 8-spored, bitunicate, fissitunicate, clavate, long pedicellate, rounded at the apex, with an ocular chamber. Ascospores 30–35× 5–8μm (x = 31.4×6.7μm, n=50), overlapping 1–2-seriate, fusiform, with narrow, acute ends, mostly curved, 7-transseptate, constricted at the central septum, cell above central septum widest, hyaline when young, becoming yellowish brown at maturity, guttulate, smooth-walled, with appendages at both ends. Asexual morph: Undetermined. Material examined: JAPAN, Oowasawa River, river bank, Kadoke, Hirosaki, Aomori, on stems of unknown herbaceous

plant, 29 July 2001, Kazuaki Tanaka KT 604 (HHUF 27309, holotype); ex-type living culture, JCM 17668; JAPAN, Sanpinai, Hirosaki, Aomori, on stems of unknown herbaceous plant, 16 September 2001, Kazuaki Tanaka KT 777 (HHUF 27310), living culture MAFF 239451. Notes: Holm and Holm (1988) introduced Lophiostoma caulium varities (var Ba-e^) based on morphological characters without a formal procedure. Tanaka and Harada (2003a) designated two strains (JCM 17668 and MAFF 239451 as Lophiostoma caulium var Bd^ based on Holm and Holm (1988). These strains grouped in the Lophiostoma clade in the present phylogenetic analysis and based on their morphological characters and phylogenetic placement we introduce these strains as a new Lophiostoma species. Lophiostoma multiseptatum shares

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similar morphological characteristics with Lophiostoma macrostomum, the type of Lophiostoma such as ascomata with a slit-like ostiole and narrowly fusiform ascospores with polar appendages, but mainly differs in having 7septate, yellowish brown ascospores. Alpestrisphaeria Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551232; Facesoffungi number: FoF00799 Etymology: In reference to where the type species was isolated (alpine soil). Isolated from soil. Sexual morph: Ascomata scattered, or clustered in small groups, erumpent, carbonaceous, black, subglobose, covered with sparse, brown, septate hyphae, ostiolate. Ostiole slit-like, central, suborbicular, papillate, periphysate, with a pore-like opening. Hamathecium comprising numerous, hyaline, filiform, septate, pseudoparaphyses, situated between and above the asci, embedded in a hyaline gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, clavate, sometime curved, pedicellate, rounded at the apex with an ocular chamber. Ascospores bi-seriate, obliquely partially overlapping, hyaline when immature, becoming brown at maturity, end cells pale brown, fusiform with narrowly rounded ends, often slightly curved, 3-septate, deeply constricted at the central septum, verruculose, lacking a mucilaginous sheath. Asexual morph: Undetermined (from Zhou et al. 2014). Type species: Alpestrisphaeria terricola (G.S. Gong) Thambugala & K.D. Hyde, comb. nov. Basionym: Trematosphaeria terricola G.S. Gong, Mycol. Progr. 13(1): 33–43 (2013) [2014] Index Fungorum Number: IF551233 Notes: Trematosphaeria terricola was introduced by Gong (in Zhou et al. 2014). Although phylogenetically this species has a close association with Lophiostoma, Zhou et al. (2014) described this species as Trematosphaeria terricola mainly based on its morphological affinities with the generic concept of Trematosphaeria. In present phylogenetic analyses, T. terricola forms an independent lineage (A14) in Lophiostomataceae (Fig. 1). Hence, we introduce a new genus Alpestrisphaeria in Lophiostomataceae to accommodate this species based on phylogeny and morphology. Biappendiculispora Thambugala, Kaz. Tanaka & K.D. Hyde, gen. nov. Index Fungorum number: IF551528; Facesoffungi number: FoF 01096; Fig. 6. Etymology: In reference to its ascospores with appendages at both ends. Saprobic on dead herbaceous stems or culms of Poaceae spp. Sexual morph: Ascomata solitary, immersed, papilla erumpent through host surface, coriaceous, reddish brown to black, subglobose, ostiolate. Ostiole rounded or slit-like, variable in shape, central, papillate, with a pore-like opening, plugged by gelatinous tissue, made up of hyaline, pseudoparenchymatous cells and occasionally lighter coloured.

Fungal Diversity (2015) 74:199–266 Fig. 5 Lophiostoma multiseptatum (HHUF 27309 and HHUF 27310). a, b. Herbarium labels. c, d. Appearance of ascomata on host surface. e. Section through ascoma f Close up of ostiole. g. Vertical sections through peridium. h. Pseudoparaphyses. i. Immature ascus. j. Mature ascus. k–n. Ascospores with terminal appendages. Scale bars: e=50μm, f=20μm, h=5μm, g, j–n=10μm

Peridium wider at the apex and thinner at the base, composed of a single stratum, with several layers of lightly pigmented to reddish brown, cells of textura angularis, cells towards the inside lighter, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising numero u s , 2 – 3 μ m w i d e , b r a n c h e d , s e p t a t e , c e l l u l a r, pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, with short, bulbous pedicel, rounded at the apex, with an ocular chamber. Ascospores overlapping uni to bi-seriate, fusiform with acute ends, 7–9-septate, constricted at the septa, cell above central septum widest, becoming pale yellowish to brown at maturity, with appendages at both ends. Asexual morph: Undetermined. Type species: Biappendiculispora japonica Thambugala, Wanasinghe, Kaz. Tanaka & K.D. Hyde, sp. nov. Index Fungorum number: IF551529; Facesoffungi number: FoF01097; Fig. 6 Etymology: Named after the country, where it was collected, Japan. Holotype: HHUF 27313 Saprobic on dead herbaceous stems or culms of Poaceae spp. Sexual morph: Ascomata 350–400 μm high, 240– 270μm diam, solitary, immersed, papilla erumpent through host surface, coriaceous, reddish brown to black, subglobose, ostiolate. Ostiole rounded or slit-like, variable in shape, up to 100–130μm long and 40–60μm wide, central, papillate, with a pore-like opening, plugged by gelatinous tissue, made up of hyaline, pseudoparenchymatous cells and occasionally lighter coloured. Peridium 15–22μm wide (x = 17.1μm, n=20), wider at the apex and thinner at the base, composed of a single stratum, with several layers of lightly pigmented to reddish brown, cells of textura angularis, cells towards the inside lighter, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising numerous, 2–3μm wide, branched, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 90–130×15–20μm (x = 108.4×18.5μm, n=40), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with short, bulbous pedicel, rounded at the apex, with an ocular chamber. Ascospores 30–36 × 6–10 μm (x = 32.4 × 7.7μm, n= 50), overlapping uni to bi-seriate, fusiform with acute ends, sometimes curved, 7–9-septate, constricted at the septa, cell above central septum widest, hyaline when young, becoming pale yellowish to brown


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Fig. 6 Biappendiculispora japonica (holotype). a. Herbarium label. b. Appearance of ascomata on host surface c. Section through ascoma d. Close up of ostiole e. Vertical section through peridium f. Pseudoparaphyses g. Immature ascus h–j. Mature asci k–n. Ascospores with terminal appendages. Scale bars: c=50μm, d=20μm, e=10μm, f=5μm, g– j=20μm, k–n=10μm

at maturity, guttulate, smooth-walled, with appendages at both ends. Asexual morph: Undetermined. Material examined: JAPAN, Sanpinai, Hirosaki, Aomori, on stems of unknown herbaceous plant, 17 July 2001, Kazuaki Tanaka KT. 573 (holotype, HHUF 27313); ex-type living culture, MAFF 239452; JAPAN: Hirakawa, river bank, Matsuzaki, Hiraka, Aomori, 18 August 2001, KT 686-1 (HHUF 27315), living culture JCM 17670; JAPAN, Campus of Hirosaki University, Hirosaki, Aomori, On culms of Dactylis glomerata (Poaceae), 1 October 2001, KT 794 (HHUF 27311), living culture JCM 17671. Notes: Biappendiculispora is introduced here as a monotypic genus based on its unique morphological characters and phylogenetic placement. The type species of this genus, Biappendiculispora japonica was originally described as Lophiostoma caulium Bvar. f^ by Tanaka and Harada (2003a). Lophiostoma multiseptatum shares similar morphological characters with B. japonica, such as fusiform, pale

yellowish to brown ascospores with narrow, acute ends, but differs in having a peridium with cells of textura prismatica and usually 7-septate ascospores. Three strains of Biappendiculispora japonica (JCM 17670, 17671, MAFF 239452) grouped as a sister clade to Pseudolophiostoma (Fig. 1). Pseudolophiostoma mainly differs from Biappendiculispora in having a peridium of textura prismatica and hyaline, broadly fusiform, 1-septate, ascospores with a mucilaginous sheath and lacking terminal appendages. Capulatispora Thambugala, Kaz. Tanaka & K.D. Hyde, gen. nov. Index Fungorum number: IF551234; Facesoffungi number: FoF00800 Etymology: In reference to capped sheath around the ascospores. Saprobic on dead twigs. Sexual morph: Ascomata solitary, scattered, immersed, coriaceous, brown, conical, with a


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flattened base, surrounded by a small blackened pseudoclypeus, ostiolate. Ostiole slit-like, central, with a slightly raised crest, with a pore-like opening. Peridium composed of a single stratum, with 4–8 layers of light pigmented to brown, flattened cells of textura angularis, cells towards the inside lighter and somewhat broad, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–1.5μm wide, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, short-pedicellate, apically narrowly rounded apex with an ocular chamber. Ascospores uni to biseriate, hyaline, becoming brownish when over mature, fusiform, with narrow, acute ends, 1-septate, upper part slightly wider, surrounded by a narrow, drawn out sheath. Sheath tips of the sheath capped, staining with blue-black ink. Asexual morph: Undetermined.

Notes: Capulatispora is introduced here in order to accommodate Lophiostoma sagittiforme Kaz. Tanaka & Hosoya (Tanaka and Hosoya 2008). Lophiostoma sagittiforme mainly differs from other genera in Lophiostomataceae in having with a flattened base, surrounded by a small blackened pseudoclypeus and ascospores with drawn-out sheaths that are capped at their tips. In our phylogenetic tree, L. sagittiforme (JCM 15100) forms a sister clade (A9) to the Guttulispora clade (A8), but these clades are morphologically distinct. Type species: Capulatispora sagittiformis (Kaz. Tanaka & Hosoya) Thambugala, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Lophiostoma sagittiforme Kaz. Tanaka & Hosoya, Sydowia 60: 134 (2008) Index Fungorum Number: IF551235; Facesoffungi number: FoF00801; Fig. 7

Fig. 7 Capulatispora sagittiformis (holotype). a. Crest-like papilla on host surface b. Ascomata produced in culture (on rice straw). c. Ascoma in longitudinal section (in lactophenol cotton blue). Note the blackened pseudoclypeus around the ostiolar opening. d. Peridium e. Pseudoparaphyses (in lactophenol cotton blue) f. Ascus g. Fissitunicate

ascus with endoascus extending from ectoascus h–k. Ascospores (i–k. in black-blue ink) l, m. Apex of ascospore appendage (in black-blue ink) n. Germinating ascospore. Scale bars. a, b=200μm, c=100μm, d, f, g, k, n= 20μm, e, h-j, l, m=5μm. b, e–i, k–m from JCM 15100 (ex-type living culture)

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Saprobic on dead twigs of Machilus japonica Siebold & Zucc. Sexual morph: Ascomata 200–250μm high, 500– 560 μm diam., solitary, scattered, immersed, coriaceous, brown, conical, with a flattened base, surrounded by a small blackened pseudoclypeus, ostiolate. Ostiole slit-like, central, with a slightly raised crest, with a pore-like opening. Peridium 15–30μm wide at sides, composed of a single stratum, with 4– 8 layers of light pigmented to brown, flattened cells (5–16×2– 10μm) of textura angularis, cells towards the inside lighter and somewhat broad, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–1.5μm wide, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 125–187.5×15–22.5μm (x = 162.1×19.1 μm, n=20), 8spored, bitunicate, fissitunicate, cylindric-clavate, short pedicellate, apically rounded, with an ocular chamber. Ascospores (33.5–)40–52(–55)×(8–)9–12.5μm (x = 46.2×10.1μm, n= 200), L/W (3.6–)4–5.2(–5.6) (x = 4.6, n=200), uni to bi-seriate, hyaline, becoming brownish when old, fusiform, with narrowed, acute ends, 1-septate, septum nearly median (0.47–0.54; x = 0.50, n=200), upper part slightly wider, smooth-walled, surrounded by a narrow, drawn out sheath. Sheath 5–32μm long (mostly 10–20μm) at both ends; tips of the sheath capped, thickened, 3–6μm high, staining with blue-black ink. Asexual morph: Undetermined. Material examined: JAPAN, Kagoshima, Isl. Yakushima, Anbou forest path, Yakusugi land, on dead twigs of Machilus japonica (Lauraceae), 22 October 2005, K. Tanaka & T. Hosoya KT 1934 (TNS-F-12453, holotype); ex-type living culture, JCM 15100. Coelodictyosporium Thambugala & K.D. Hyde, gen. nov. Index Fungorum Number: IF551286; Facesoffungi number: FoF00802 Etymology: In reference to a new coeleomycetous genus with Dictyosporium-like spores. Saprobic on dead branches in terrestrial habitats. Sexual morph: Ascomata solitary or gregarious, immersed, coriaceous, black, globose to subglobose, ostiolate, apex well-developed. Ostiole crest-like, central, broadly papillate, with a pore-like opening. Peridium wider at the apex and thinner at the base, composed of two strata, outer stratum comprising dark brown to black, somewhat flattened cells of textura angularis, fusing and indistinguishable from the host tissues, and inner stratum comprising 2–3 layers of brown to lightly pigmented cells of textura angularis. Hamathecium comprising 1.5–3μm wide, numerous, aseptate, unbranched, cellular pseudoparaphyses, anastomosing above the asci, embedded in a gelatinous matrix. 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicel, apically rounded, with an ocular chamber. Ascospores uni to bi-seriate, partially overlapping, yellowish to brown, ellipsoid to fusiform, muriform, 5–6 transversely septate, with 1–2 vertical septa, deeply constricted at the middle septum, upper part slightly wider, with


terminal appendages and surrounded by a thick mucilaginous sheath. Asexual morph: Colonies growing on dead branches. Conidiomata pycnidial, on the upper surface of stems, solitary, scattered, superficial, globose to subglobose, black. Peridium thin at the apex, base a single layer, composed of brown to black thin-walled cells of textura angularis. Conidiogenous cells holoblastic, integrated, smooth, brown, producing a branched conidium at the apex, cup-shaped or doliiform. Conidia uniformly medium brown, complanate, dictyosporous, regularly consisting of 6–8 rows of cells, each row comprising 5–7 cells (asexual morph description follows Lophiostoma pseudodictyosporium in Liu et al. 2015). Type species: Coelodictyosporium pseudodictyosporium (Qing Tian, Camporesi & K.D. Hyde) Thambugala & K.D. Hyde, comb. nov. Index Fungorum Number: IF551236 Basionym: Lophiostoma pseudodictyosporium Qing Tian, Camporesi & K.D. Hyde, in Liu et al., Fungal Diversity 72(1): 114 (2015) Notes: Lophiostoma pseudodictyosporium was introduced by Liu et al. (2015) as an asexual species in Lophiostomataceae mainly based on its phylogenetic placement in Lophiostomataceae. In the present phylogenetic analysis L. pseudodictyosporium forms a separate subclade (A3), which is sister to Neotrematosphaeria (A4) along with a new strain MFLUCC 13–0351. Coelodictyosporium dictyosporium is morphologically similar with species of Dictyosporium in having brown applanate conidia. Conidia formation of D i c t y o s p o r i u m i s d ir e c t l y o n m y c e l i a , w h i le i n Coelodictyosporium pseudodictyosporium this occurs in pycnidia (Liu et al. 2015). Our new specimen MFLU 14– 0586 (MFLUCC 13-0351) was collected from Italy on Spartium junceum and this species differs from other Lophiostoma species in having a broadly papillate ostiole, yellowish to brown, muriform ascospores with small apical appendages. Because of these unusual characters we introduce a new genus to accommodate these sexual and asexual species. Coelodictyosporium muriforme Thambugala, E. Camporesi & K.D. Hyde, sp. nov. Index Fungorum Number: IF551237; Facesoffungi number: FoF00803; Fig. 8 Etymology: Referring to its muriform ascospores. Holotype: MFLU 14-0586 Saprobic on dead branches of Spartium junceum L. Sexual morph: Ascomata 400–700μm high, 350–550μm diam. (x = 490×425μm, n=5), solitary or gregarious, immersed, coriaceous, black, globose to subglobose, ostiolate, apex well-developed. Ostiole crest-like, central, broadly papillate, with a pore-like opening. Peridium 25–60μm wide (x = 40μm, n= 15), wider at the apex and thinner at the base, composed of two strata, outer stratum comprising dark brown to black, somewhat flattened cells of textura angularis, fusing and indistinguishable from the host tissues, and inner stratum


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Fig. 8 Coelodictyosporium muriforme (holotype). a, b. Appearance of ascomata on the host tissue c. Section through ascoma d. Ostiole (note the welldeveloped apex) e. Section of the peridium f. Pseudoparaphyses g Immature ascus h–i Mature bitunicate asci j–l. Ascospores (note the small appendages in k) m. Ascospore stained in Indian ink. Scale bars: c-d=100μm, e, f =25μm, g–i=50μm, j–m=25μm

comprising 2–3 layers of brown to lightly pigmented cells of textura angularis. Hamathecium comprising 1.5–3μm wide, numerous, aseptate, unbranched, cellular pseudoparaphyses, anastomosing above the asci, embedded in a gelatinous matrix. Asci 85–135×12–17μm (x = 119×14.5μm, n=20), 8spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicel, apically rounded, with an ocular chamber. Ascospores 17–23.5×6.5–9μm (x = 20×7.4μm, n=25), uni to bi-seriate, partially overlapping, yellowish to brown, ellipsoid to fusiform, muriform, 5–6 transversely septate, with 1–2 vertical septa, deeply constricted at the middle septum, upper part slightly wider, with small, pointed appendages at both ends and surrounded by a thick mucilaginous sheath. Asexual morph: Undetermined.

Culture characteristics: Ascospores germinating on PDA within 18 h and germ tubes produced from several septa. Colonies on PDA reaching 10 mm diam. after 4 weeks at 25 °C, flat to effuse, surface white, reverse yellowish-orange, dense, aerial, surface smooth, with entire to slightly undulate edge. Material examined: ITALY, Province of Forlì-Cesena [FC], Montebello, Modigliana, on dead branches of Spartium junceum (Fabaceae), 20 October 2012, Erio Camporesi IT 839 (MFLU 14-0586, holotype); ibid., (HHUF 30458, isotype); ex-type living culture, MFLUCC 13-0351, ICMP; ibid., Passo delle Forche, Galeata, on dead branches of Spartium junceum (Fabaceae), 28 February 2013, Erio Camporesi IT 839-2 (MFLU 15-1122, paratype); ibid.,

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Marsignano, Predappio, on dead branches of Spartium junceum (Fabaceae), 14 March 2013, Erio Camporesi IT 839-3 (MFLU 15-1123, paratype). Guttulispora Thambugala, Qing Tian & K.D. Hyde, gen. nov. Index Fungorum number: IF551238; Facesoffungi number: FoF00804 Etymology: In reference to its guttulate ascospores. Saprobic on dead branches in terrestrial habitats. Sexual morph: Ascomata solitary or scattered, immersed, coriaceous to carbonaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole central, papillate, with a small crest-like apex and an irregular pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium composed of a single stratum with dark to reddish brown, thick-walled cells of textura angularis, cells towards the inside lighter, at the outside, darker, somewhat compressed, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2 μm wide, septate, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 8spored, bitunicate, fissitunicate, cylindrical, short pedicellate, apically rounded with an ocular chamber. Ascospores uniseriate, partially overlapping, hyaline when immature and becoming brown when mature, ellipsoid to fusiform, 3-septate, constricted at the each septa, upper part slightly wider, guttulate at each cell, lacking a mucilaginous sheath. Asexual morph: Undetermined. Type species: Guttulispora crataegi Qing Tian, Thambugala, Camporesi & K.D. Hyde, sp. nov. Index Fungorum Number: IF551239; Facesoffungi number: FoF00805; Fig. 9 Etymology: Referring to the host genus Crataegus. Holotype: MFLU 14-0584 Saprobic on a dead and branches of Crataegus sp. Sexual morph: Ascomata 175.5–363×152.5–313.5μm diam. (x = 288.5×226μm, n=10), solitary or scattered, immersed, coriaceous to carbonaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole central, papillate, with a small crest-like apex and an irregular pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 28–46.5μm wide (x = 38μm, n= 12), composed of a single stratum, with dark to reddish brown, thick-walled cells of textura angularis, cells towards the inside lighter, at the outside, darker, somewhat compressed, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2 μm wide, septate, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 45–130×7.5–11μm (x = 83×9.5μm, n=10), 8-spored, bitunicate, fissitunicate, cylindrical, short-pedicellate, apically rounded with an ocular chamber. Ascospores 14.5–18.5×5.5–7μm (x = 16×6μm, n = 10), uniseriate, partially overlapping, hyaline when


immature and becoming brown when mature, ellipsoid to fusiform, 3-septate, constricted at the each septa, upper part slightly wider, guttulate at each cell, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined. Culture characteristics: Ascospores germinating on PDA within 18 h and producing germ tubes from several septa. Colonies on PDA reaching 20 mm diam. after 4 weeks at 25 °C, effuse to raised, surface grey, reverse greenish-black, dense, with entire to slightly undulate, greyish green edge, producing a yellowish pigment in the culture. Material examined: ITALY, Province of Forlì-Cesena [FC], Marsignano - Predappio, on dead branch of Crataegus sp. (Rosaceae), 15 March 2013, Erio Camporesi IT 232 (MFLU 14–0584, holotype); ibid., (HHUF 30457, isotype), ex-type living culture, MFLUCC 13-0442, ICMP; ibid., 14 May 2013, Erio Camporesi IT 232-2 (MFLU 14-0585, paratype), living culture, MFLUCC 14-0993, ICMP. ibid., 15 April 2013, Erio Camporesi IT 232-3, IT 232-4, IT 232-5 (MFLU 15-1124, 151125, 15-1126, paratypes). Notes: Guttulispora crataegi is distinct from other species in Lophiostomataceae in having small, brown, guttulate, 3septate ascospores, constricted at each septa. In the phylogenetic analysis (Fig. 1) two strains of G. crataegi (MFLUCC 13-0442 and MFLUCC 14-0993), including the ex-type strain, formed a sister group to the new genus Capulatispora which is characterized by coriaceous, conical ascomata surrounded by a small blackened pseudoclypeus, a 4–8 layered, light brown peridium and hyaline ascospores, surrounded by a narrow mucilaginous sheath. Therefore, a new genus Guttulispora is introduced here to accommodate G. crataegi. Lophiohelichrysum Dayarathne et al., in Ariyawansa et al., Fungal Diversity (in press) Notes: Ariyawansa et al. (2015) introduced Lophiohelichrysum as a new genus in Lophiostomataceae based on its unique morphological characteristics and phylogenetic placement to accommodate Lophiohelichrysum helichrysi Dayarathne et al. Lophiohelichrysum is characterized by coriaceous ascomata with slit-like ostioles and 1-septate, assymetrical, hyaline to lightly pigmented ascospores. The ex-type strain of Lophiohelichrysum helichrysi (MFLUCC 15-0701) grouped as aseparate clade (A7) in the phylogenetic tree resulted from the present study (Fig. 1). Lophiopoacea Ariyawansa, Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551240; Facesoffungi number: FoF00806 Etymology: In reference to Lophiostoma-like genus and its Poaceae host. Saprobic on Poaceae species in terrestrial habitats. Sexual morph: Ascomata scattered to gregarious, immersed, coriaceous to carbonaceous, dark brown to black, surrounded by a small blackened pseudoclypeus, subglobose to conical,


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Fig. 9 Guttulispora crataegi (holotype). a–c. Appearance of ascomata on the host surface d. Section through ascoma (note the small crest-like apex) e. Ostiole. f. Section through peridium g. Squash mount of numerous, crowded asci h. Pseudoparaphyses i. Immature ascus j, k. Mature asci l. Apex of ascus with an ocular chamber m. Germinating ascospore n, o. Immature hyaline ascospores p, q. Mature brown ascospores r. Colony on PDA from above. Scale bars: b=1000μm, c= 100μm, d, g=50μm, e, f=20μm, i–l=5μm, h, m–p=10μm

ostiolate. Ostiole slit-like, central, with a reduced crest and a pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium wider at the apex, and thinner at the base, comprising several layers of lightly pigmented to dark brown, cells of textura angularis, cells towards the inside lighter and somewhat broad, at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1.5–2.5μm wide, septate, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix.

Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, pedicellate, rounded at the apex, with an indistinct ocular chamber. Ascospores uniseriate to bi-seriate, overlapping, hyaline, fusiform with narrow, acute ends, 1–5-septate, constricted at the central septum, with or without a mucilaginous sheath. Asexual morph: Undetermined. Type spec ie s : Lophiop oacea pa ramacrostoma Ariyawansa, Thambugala, Camporesi & K.D. Hyde, sp. nov. Index Fungorum Number: IF551241; Facesoffungi number: FoF00807; Fig. 10

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Fig. 10 Lophiopoacea paramacrostoma (holotype). a. Appearance of ascomata on host surface b. Section through ascoma c, d. Peridium. note the ostiole in c e. Pseudoparaphyses f. Immature ascus g. Mature ascus with ascospores h–k. Ascospores. Scale bars: b=100μm, c, d= 50μm, e=10μm, f, g=20μm, hk=20μm

Etymology: Referring to the host genus Arundo. Holotype: MFLU 14-0590 Saprobic on Poaceae species in terrestrial habitats. Sexual morph: Ascomata 220–310μm high×220–375μm diam. (x = 267×289μm, n=5), scattered to gregarious, immersed, coriaceous, dark brown to black, surrounded by a small blackened

pseudoclypeus, subglobose to conical, ostiolate. Ostiole slitlike, central, with a reduced crest and a pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 30–65μm wide (x = 46μm, n=15), wider at the apex, and thinner at the base, composed of two strata, outer stratum comprising several


layers with dark brown to black, somewhat flattened cells of textura angularis, fusing and indistinguishable from the host tissues, inner stratum comprising several layers with lightly pigmented cells of textura angularis. Hamathecium comprising 1.5 –2.5 μm wide, septate, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 75–105×9–13.5μm (x = 86×11.5μm, n=15), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicel, rounded at the apex, with an indistinct ocular chamber. Ascospores 35–40×4.8–6.7μm (x = 37.7×5.6μm, n=20), uniseriate to bi-seriate, overlapping, hyaline, fusiform with narrow, acute ends, 1-septate, euseptate, constricted at the septum, region above central septum widest, smooth-walled, guttulate, lacking a mucilaginous sheath. Asexual morph: Undetermined. Culture characteristics: Ascospores germinating on PDA within 24 h and producing germ tubes from both ends. Colonies on PDA growing slowly, reaching 52 mm diam. after 10 days at 25 °C, effuse, circular, whitish grey, dense, with entire to slightly undulate edge. Material examined: ITALY, Province of Forlì-Cesena [FC], Monte Vescovo, on dead Arundo pliniana Turra (Poaceae), 20 December 2011, Erio Camporesi E3 (MFLU 14-0590, holotype); ibid., (HHUF 30461, isotype), ex-type living culture, MFLUCC 11-0463, ICMP 20759. Notes: We introduce a new genus Lophiopoacea in Lophiostomataceae based on both morphological characters together with multi-gene phylogeny of the new species Lophiopoacea paramacrostoma and Lophiostoma winteri (Sacc.) G. Winter. Lophiopoacea and Lophiostoma are morphologically similar genera, but they are phylogenetically different (Fig. 1). In the phylogenetic tree the ex-type strain of L. paramacrostoma clustered with two putatively named strains of Lophiostoma winteri (JCM 17648, MAFF 239454) as a distinct group. Therefore, Lophiopoacea winteri is introduced for Lophiostoma winteri and accepted as second species of Lophiopoacea. Lophiopoacea winteri differs from L. paramacrostoma in having a peridium with brown to reddish brown, flattened cells of textura angularis and 5-septate ascospores, with a thin mucilaginous sheath. Lophiopoacea paramacrostoma shares similar m orphological traits with Lophiostoma macrostomum, the type species of Lophiostoma. Both have ascomata with slit-like ostioles and 1-septate, hyaline, fusiform ascospores with narrow, acute ends. L o p h i o p o ac e a p a r a m a c ro s t o m a d i ffe r s fr o m Lophiostoma macrostomum in having a reduced crest, a peridium comprising two strata with hyaline to brown, large cells of textura angularis, wider pseudoparaphyses and larger, fusiform ascospores. Lophiopoacea is therefore proposed for the subclade A6 (Fig. 1). Lophiopoacea winteri (Sacc.)Thambugala, Jayasiri, Kaz. Tanaka & K.D. Hyde, comb. nov.

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Basionym: Lophiotrema winteri Sacc., Syll. fung. (Abellini) 2: 688 (1883) ≡ Lophiostoma winteri (Sacc.) G. Winter, Rabenh. Krypt.Fl., Edn 2 (Leipzig) 1.2: 297 (1885) Index Fungorum number: IF551530; Facesoffungi number: FoF00808; Fig. 11 Saprobic on stem of Polygonum sp. Sexual morph: Ascomata 290–320μm high, 310–330μm diam. (x = 300× 314 μm, n = 10), solitary or scattered, immersed, black, subglobose to globose, coriaceous to carbonaceous, ostiolate. Ostiole slit-like, central, rounded, with a pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 35–41μm wide (x = 39μm, n=10), composed of a single stratum, with several layers of brown to reddish brown, flattened cells of textura angularis, cells towards the inside lighter and at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1.5–2μm wide, numerous, filamentous, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 87– 102 × 10–13 μm (x = 90 × 12.5 μm, n = 15), 8-spored, bitunicate, fissitunicate, cylindrical, with a short, bulbous pedicel, rounded at the apex, with an ocular chamber. Ascospores 30–33×5–5.9μm (x = 31.5×5.3μm, n=20), bi-seriate, overlapping, hyaline, fusiform, with narrow, acute ends, usually 5septate, constricted at the central septum, cell above central septum widest, filled with small guttules when immature, surrounded by a thin, sheath, drawn out at the ends. Asexual morph: Undetermined. Material examined: JAPAN: Toyohira River, river bank, Sapporo, Hokkaido, on stem of Polygonum sp., 2 September 2001, K. Tanaka KT. 764 (HHUF 27316), living culture MAFF 239454; JAPAN: Akaiwa, Funadomari, Isl. Rebun, Hokkaido, on stems of an unknown plant, 31August 2001, KT 740 (HHUF 27317), living culture JCM 17648. Neotrematosphaeria Thambugala, Kaz. Tanaka & K.D. Hyde, gen. nov. Index Fungorum number: IF551242; Facesoffungi number: FoF00809 Etymology: The generic epithet, neo (Lat., new), refers to the similarity to Trematosphaeria. Saprobic on dead twigs in freshwater habitats. Sexual morph: Ascomata solitary, semi-immersed or slightly erumpent, coriaceous to carbonaceous, black, subglobose, ostiolate. Ostiole rounded, central, broadly papillate, with a crest-like apex and a pore-like opening. Peridium broad at sides, poorly developed at the base, composed of a single stratum, with several layers of dark brown to black, cells of textura prismatica, cells towards the inside lighter and at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–1.5μm wide, filiform, cellular, pseudoparaphyses, branching above and between asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate,

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Fig. 11 Lophiopoacea winteri (HHUF 27316). a Herbarium material. b-c Appearance of ascomata on the host tissue d. Section through ascoma e. Section through peridium f. Pseudoparaphyses g, h Immature asci i Mature bitunicate ascus j– m. Ascospores (m. in black-blue ink). Scale bars: d=100μm, e= 50μm, f, j–m=10μm, g–i=30μm

fissitunicate, cylindric-clavate, with a long pedicel, rounded at the apex, with an ocular chamber. Ascospores uni to bi-seriate, partially overlapping, pale yellowish to olivaceous brown, except at the pale end cells, fusiform, mostly curved, 5(–7)-septate, slightly constricted at the septa, cell above central septum widest, with appendages at both ends. Asexual morph: See notes. Notes: Although described as Trematosphaeria biappendiculata (Tanaka et al. 2005a), several morphological features of this species indicate that it is a member of Lophiostomataceae. However, T. biappendiculata differs from the other species in Lophiostomataceae by having a peridium with 15–25 layers of dark brown to black, cells of

textura prismatica, and pale yellowish to olivaceous brown, 5(–7)-septate ascospores with globose to conical terminal appendages. Tanaka et al. (2005a) reported the asexual morph of this fungus as Bpleurophomopsis^-like. It should however, be regarded as a spermatial state because the Bconidia^ produced in culture did not germinate. In this study, Trematosphaeria b i a p p e n d i c u l a t u m ( K T C 11 2 4 ) c l u s t e r e d i n t h e Lophiostomataceae as a sister clade to the Coelodictyosporium clade (A3, Fig. 1). Morphologically, Trematosphaeria biappendiculata is close to T. lineolatispora K.D. Hyde and T. pachycarpa (Sacc. & El. Marchal) Shoemaker & C.E. Babc., but T. lineolatispora the has striate ornamentation in ascospore surface and is a marine species


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Fig. 12 Neotrematosphaeria biappendiculata (paratype and holotype of Neotrematosphaeria biappendiculata). a, b. Appearance of ascomata on host surface c–e. Ascomata in longitudinal section (d. in lactophenol cotton blue) f. Peridium at side. g. Pseudoparaphyses h, i. Asci j. Fissitunicate ascus with endoascus extending from ectoascus k–p. Ascospores (k–m. in black-blue

ink) q. Germinating ascospore r. Spermatia. Scale bars. a=1 mm, b=200μm, c–e=100μm, f, g, j, q=10μm, h, i, k–p, r=5μm. a–d, k, l from HHUF 28017 (paratype); e–j, m–q from HHUF 28026 (holotype); r from MAFF 239296, JCM 12850 (ex-paratype living culture)

while T. pachycarpa has median central septum and graminicolous habit (Shoemaker and Babcock 1989). Type species: Neotrematosphaeria biappendiculata (Kaz. Tanaka, Y. Harada & M.E. Barr) Thambugala, Kaz. Tanaka & K.D. Hyde, comb. nov.

Basionym: Trematosphaeria biappendiculata Kaz. Tanaka, Y. Harada & M.E. Barr, Fungal Diversity 19: 149 (2005) Index Fungorum Number: IF551243; Facesoffungi number: FoF00810; Fig. 12

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Saprobic on dead twigs in freshwater habitats. Sexual morph: Ascomata 450–860μm high, 750–980μm diam., solitary, semi-immersed or slightly erumpent, coriaceous to carbonaceous, black, subglobose, ostiolate. Ostiole rounded, central, broadly papillate, with a crest-like apex and a pore-like opening. Peridium 55–100μm wide at sides, poorly developed at the base, composed of a single stratum, with 15–25 layers of dark brown to black, cells of textura prismatica, cells towards the inside lighter and at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–1.5μm wide, filiform, cellular, pseudoparaphyses, branching above and between asci, embedded in a gelatinous matrix. Asci 112.5–152.5×15.5–20μm (x = 133.6×17.6μm, n=27), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicel, rounded at the apex, with an ocular chamber. Ascospores (28–)31–37(–40.5)×(8–)9–10.5(–12) μm (x = 33.6×9.8μm, n=172), L/W (3–)3.2–3.7(–3.9) (x = 3.4, n= 172), uni to bi-seriate, partially overlapping, pale yellowish to olivaceous brown, except at the pale end cells, fusiform, mostly curved, 5(–7)-septate, slightly constricted at the septa, cell above central septum widest, with a primary submedian septum (0.50–0.54; x = 0.52, n=161), smooth-walled, with appendages at both ends. Appendages globose to conical, 0.5–3μm diam., staining blue-black in Indian ink. Asexual morph: Bpleurophomopsis^-like. Material examined: JAPAN, Aomori, Hirosaki, Aoki, Mohei Pond, on submerged wood, 3 May 2003, K. Tanaka & N. Asama, KT 1124 (HHUF 28026, holotype); extype living culture, MAFF 239297, JCM 12849; Aomori, Hirosaki, Matsuzaki, Serisawa Park (pond), on submerged wood, 7 December 2002, K. Tanaka & N. Asama, KT 975 (HHUF 28017, paratype); ex-paratype living culture MAFF 239296, JCM 12850. Paucispora Thambugala, Kaz. Tanaka & K.D. Hyde, gen. nov. Index Fungorum number: IF551244; Facesoffungi number: FoF00811 Etymology: In reference to having fewer than eight ascospores in the ascus. Saprobic on dead twigs in terrestrial habitats. Sexual morph: Ascomata scattered, immersed, coriaceous, black, globose to subglobose, ostiolate. Ostiole slit-like, central, with a crest-like papilla, with an irregular, pore-like opening, plugged by hyaline, filamentous hyphae, and occasionally lighter coloured. Peridium wider at the apex, and thinner at the base, composed of several layers with brown to black cells of textura angularis, cells towards the inside lighter and at the outside, darker, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2μm wide, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 2–4-spored, bitunicate, fissitunicate, cylindrical-clavate, with a long pedicel apically rounded with an ocular chamber. Ascospores uniseriate,


partially overlapping, hyaline to reddish brown, ellipsoidal, 1–3-septate, constricted at the septum, lacking a mucilaginous sheath. Asexual morph: Undetermined. Notes: Lophiostoma quadrisporum was introduced by Hirayama and Tanaka (2011) based on misidentified material of Lophiotrema nucula (HHUF 27321; Tanaka and Harada 2003b). Hirayama and Tanaka (2011) showed phylogenetic affinity of this species with Lophiostoma rather than with Lophiotrema. In the phylogenetic analysis (Fig. 1), L. quadrisporum forms a well-supported clade (B2) with L. versicolor (MAFF 244508, Hirayama et al. 2014). Lophiostoma quadrisporum and L. versicolor have distinctive 2–4-spored asci and ellipsoidal, 1–3-septate ascospores. Ascomata with slit-like ostioles, and crest-like papilla, cylindrical-clavate asci and ellipsoidal ascospores suggest that these taxa are members of Lophiostomataceae, but they also have unusual features for the genus, such as 2(–4)-spored asci and hyaline to reddish brown ascospores (Hirayama et al. 2014). Because of the unique morphological traits and phylogenetic inference, we introduce Paucispora to accommodate L. quadrisporum and L. versicolor. Type species: Paucispora quadrispora (K. Hiray. & Kaz. Tanaka) Thambugala, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Lophiostoma quadrisporum K. Hiray. & Kaz. Tanaka, Mycoscience 52(6): 407 (2011) Index Fungorum number: IF551245; Facesoffungi number: FoF00812; Fig. 13 Saprobic on dead twigs of Liriodendron tulipifera L. Sexual morph: Ascomata 300–360μm high, 300–435μm diam., scattered, immersed, coriaceous, black, globose to subglobose, ostiolate. Ostiole slit-like, central, with a crestlike papilla, with an irregular, pore-like opening, plugged by hyaline, filamentous hyphae, and occasionally lighter coloured. Peridium 10–30μm wide at sides, composed of two strata, outer stratum wider, comprising several layers with black, somewhat flattened cells of textura angularis fusing and indistinguishable from the host tissues and inner stratum comprising 2–4 layers of lightly pigmented to hyaline cells of textura angularis. Hamathecium comprising 1–2μm wide, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci (70–)80–110(–120)×(8–)9–11.5μm (x = 98.9×10μm, n= 28), 4-spored, bitunicate, fissitunicate, cylindrical-clavate, with a long pedicel (10–34μm long; x = 27μm, n=12), apically rounded with an ocular chamber. Ascospores (19–)20– 24(–26) × 6–8(–9.5) μm (x = 22 × 7.5 μm, n = 106), L/W (2.4–)2.7–3.2(–3.6) (x = 2.9, n=106), uniseriate, partially overlapping, hyaline, ellipsoidal, 1-septate, becoming 3septate at germination, constricted at the septum, with two distinct large guttules in each cells, smooth-walled, laking a mucilaginous sheath. Asexual morph: Undetermined. Material examined: JAPAN, Iwate, Morioka, Ueda, Iwate University Garden, on twigs of Liriodendron tulipifera


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Fig. 13 Paucispora quadrispora (holotype). a, b. Appearance of ascomata on host surface. c, d. Ascomata in longitudinal section (in lactophenol cotton blue) with mainly immersed crest-like papilla e. Peridium (in lactophenol cotton blue) f. Pseudoparaphyses g, h. Asci i–m. Ascospores n. Germinating ascospore. Scale bars. a, b= 200μm, c, d=50μm, e–h, i–n= 10μm, m from MAFF 239455 (ex-type living culture)

(Magnoliaceae), 11 January 2002, Y. Harada KT 843 (HHUF 27321, holotype); ex-type living culture, MAFF 239455; Fukuoka, Higashi-Park, on woody plant, 25 March 2012, K. Hirayama KH 448 (HHUF 30455). Paucispora versicolor (K. Hiray. & Kaz. Tanaka) Thambugala, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Lophiostoma versicolor K. Hiray. & Kaz. Tanaka, Mycosphere 5 (3): 414 (2014) Index Fungorum number: IF551246 Facesoffungi number: FoF00813; Fig. 14 Saprobic on dead twigs of Acer sp. Sexual morph: Ascomata 330–350μm high, 360–400μm diam., scattered to gregarious, semi-immersed, coriaceous, black, globose to subglobose, coriaceous, ostiolate. Ostiole slit-like, central, with a crest-like apex, broadly papillate, with a pore-like opening. Peridium 20–27.5μm wide at sides, wider at the apex, and thinner at the base, composed of a single stratum, with 5–7 layers of brown to lightly pigmented cells of textura angularis, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2μm wide, numerous, septate, branched pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 60–67.5(–77.5)× 10–12.5(–15) μm, 2(–4)-spored, bitunicate, fissitunicate, cylindrical-clavate, with a short pedicel, apically rounded, with an ocular chamber. Ascospores 18–23×6–9μm (x = 20.6×

6.9 μm, n = 90), L/W (2.3–)2.5–3.4(–3.7) (x = 3, n = 90), uniseriate, partially overlapping, brown to reddish brown, except pale end cells, ellipsoidal, straight, 3-septate, constricted at the central septum, with a distinct guttule in each cell, verruculose, lacking a mucilaginous sheath. Asexual morph: Undetermined. Material examined: JAPAN, Mie, Tsu, Mie University garden, on twigs of Acer sp. (Sapindaceae), 30 May 2008, K. Tanaka & H. Yonezawa KH 110 (HHUF 30448, holotype); ex-type living culture, MAFF 244508. Platystomum Trevis., Bull. Soc. R. Bot. Belg. 16: 16 (1877) Facesoffungi number: FoF00814 Saprobic on woody substrates in terrestrial habitats. Sexual morph: Ascomata solitary or gregarious, immersed to semiimmersed, papilla erumpent through host surface, coriaceous to carbonaceous, dark brown to black, globose to subglobose or conical, ostiolate. Ostiole slit or slot-like, central, papillate, with or without a crest, opening by an apical, lysigenous pore or dehiscence, usually plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium broad at the apex and thinner at the base, comprising two strata with several layers of brown or lightly pigmented to hyaline cells of textura angularis to textura prismatica, fusing and indistinguishable from the host tissues. Hamathecium

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Fig. 14 Paucispora versicolor (holotype). a, b. Appearance of ascomata on host surface. Note the crest-like papilla surrounding ostiole. c-e. Ascomata in longitudinal section f. Peridium at the side g. Peridium around neck h. Pseudoparaphyses i, j. Asci k–o Ascospores p.

Germinating ascospore. Scale bars: a, b=500μm, c–e=100μm, f, g= 20μm, h-j, p=10μm, k-o =5μm; n, o from MAFF 244508 (ex-type living culture)

comprising 1–3μm wide, numerous, branch or unbranched, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 8spored, bitunicate, fissitunicate, cylindric-clavate, pedicellate, apically rounded, with an ocular chamber. Ascospores uni to bi-seriate, partially overlapping, hyaline when immature, becoming brown to dark brown when mature, ellipsoidal to fusiform, muriform, 3–8 transversely septate, with 1–7 vertical septa, constricted at the central septum, lacking a mucilaginous sheath. Asexual morph: Coelomycetous. Conidiomata pycnidial, phoma-like, solitary, gregarious, dark brown to black, slightly erumpent, papillate, with a sphaerical mass of slimy conidia oozing out at ostiolar tip. Conidiomata wall multi-layered, with 3–4 outer layers of brown-walled cells of textura angularis, with inner most layer thin, hyaline. Conidiophores short, septate, sparsely branched, formed from the inner most layer of pycnidium wall. Conidiogenous cells phialidic, cylindrical, hyaline, flexuous, smooth, with short

collarette. Conidia hyaline, aseptate, straight to curved, ellipsoidal, with rounded ends, thin-walled, smooth, numerous. Type species: Platystomum compressum (Pers.) Trevis., Bull. Soc. R. Bot. Belg. 16: 16 (1877) Facesoffungi number: FoF 00815; Figs. 15 and 16 Reference specimen: MFLU 14-0589 = Lophiostoma lojkanum (Sacc.) Mussat, in Saccardo, Syll. fung. (Abellini) 15: 198 (1900) Saprobic on woody substrates in terrestrial habitats. Sexual morph: Ascomata 400–950μm high, 350–950μm diam. (x = 654×606μm, n=15), solitary or gregarious, immersed, papilla erumpent through host surface, coriaceous to carbonaceous, black, globose to subglobose, ostiolate. Ostiole slit-like, central, papillate, with a reduced crest and a pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 50–130μm wide (x = 85μm, n=15), wider at the apex and thinner at the base, comprising two strata, outer layer composed of dark brown-


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Fig. 15 Platystomum compressum (Reference specimen). a, b. Appearance of ascomata on the host tissue c, d. Sections through ascomata e. Ostiole f. Section through peridium g. Pseudoparaphyses h. Immature ascus i–j. Mature bitunicate asci k. Ascospores l. Germinating ascospore m. Colony on PDA. Scale bars: c-d= 300μm, e–f=50μm, g–k=25μm, i=10μm

walled, flattened cells of textura prismatica, fusing and indistinguishable from the host tissues and the inner layers with 6– 8 layers of brown to lightly pigmented cells of textura angularis. Hamathecium comprising 1–3μm wide, numerous, branched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 76– 160×8–15μm (x = 126×13μm, n=15), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicel, apically rounded with an ocular chamber. Ascospores 17–32.5×7– 9.5μm (x = 22.5×7.9μm, n=30), uniseriate, partially overlapping, hyaline when immature, becoming brown to dark brown when mature, ellipsoidal to fusiform, muriform, (4−)5–7 transversely septate, with 1–3 vertical septa, constricted at the central septum, guttulate, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined.

Culture characteristics: Ascospores germinating on PDA within 18 h and producing germ tubes from several septa. Colonies on PDA growing slowly, attaining 12 mm diam. after 21 days at 25–30 °C, white at first, after 1 to 2 weeks becoming ashy-white, flat to effuse, raised in the middle, velvety, dense, surface smooth, with entire to slightly undulate edge. Material examined: HUNGARY, Somogy, Kaposvàr in Ungarn, Lojka, 1872 (S-F7232, holotype of Lophiostoma lojkanum Rehm.); ITALY, Province of Forlì-Cesena [FC], Strada San Zeno, Galeata, on dead branch of Salix sp. (Salicaceae), 1 December 2012, Nello Camporesi IT 932 (MFLU 14-0589, HHUF 30462, reference specimen of Platystomum compressum designated here); living culture, MFLUCC 13-0343, ICMP.

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Fig. 16 Platystomum compressum (S-F7232 holotype of Lophiostoma lojkanum) a. Herbarium material b,c. Appearance of ascomata on the surface or immersed in the host d. Vertical section of ascoma e. Ostiole f. Peridium g. Immature ascus h–k. Asci with ascospores l–n. Ascospores o. Pseudoparaphyses. Scale bars: b=1000μm, c=200μm, d= 100μm, e=50μm, f=20μm, g– k=10μm, l–o=5μm

Notes: Cesati and De Notaris (1863) transferred and accommodated Sphaeria compressa Pers. in Lophiostoma (Lophiostomataceae) based on immersed to slightly erumpent ascomata with a cylindrical papilla. Lophiostoma compressum was transferred to several different genera, but Zhang et al. (2009, 2011) placed L. compressum in Lophiostoma based on its phylogenetic placement. Platystomum was introduced by Trevisan (1877) and is typified by Platystomum compressum (Pers.) Trevis. Platystomum compressum is listed as a

synonym of Lophiostoma compressum (Pers.) Ces. & De Not. in Index Fungorum (2015). However, Mugambi and Huhndorf (2009) showed in their combined phylogenetic analysis, a putatively named strains of P. compressum grouped with Pseudotrichia guatopoensis Huhndorf and Xenolophium guianense Huhndorf in a separate clade from Lophiostomataceae, which may correspond to the family Platystomaceae (Mugambi and Huhndorf 2009; Hyde et al. 2013). However, in this phylogenetic analysis (Fig. 1) the


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Fig. 17 Platystomum actinidiae (holotype). a. Herbarium material b. Appearance of ascomata on the host tissue c. Section through ascoma d. Ostiole e. Section through peridium f. Pseudoparaphyses g, h Immature asci i Mature bitunicate ascus j– m. Ascospores. Scale bars: c =150μm, d =100μm, e, g–I= 30μm, f=10μm, j–m=5μm

strain (MFLUCC 13-0343) from new collection of P. compressum grouped in Lophiostomataceae with other new species of Platystomum (MFLUCC 14-0925, 15-0632, 15-0633) and the putatively named strains of L. compressum (KT 521, JCM 13125 and IFRD 2014), Lophiostoma triseptatum Peck (SMH 5287 and SMH 2591) and Lophiostoma viridarium Cooke (IFRDCC 2090) in a same subclade (A1). We observed and illustrated the specimens attached to KT 521, JCM 13125 strains and the strains of L. triseptatum and L. viridarium in GenBank are named without attached voucher material. Therefore, it is impractical to confirm their characters. Re-collection, epitypification or a reference specimen (Ariyawansa et al. 2014b) with molecular data are essential for the type species of L. viridarium and L. triseptatum to establish their placement in Platystomum.

We accept Platystomum in Lophiostomataceae as a distinct genus and introduce four new species in Platystomum based on new collections and two strains (KT 521 and JCM 13125). Platystomum actinidiae Thambugala, Jayasiri, Kaz. Tanaka & K.D. Hyde, sp. nov. Index Fungorum Number: IF551531; Facesoffungi number: FoF00816; Fig. 17 Etymology: Referring to the host genus Actinidia. Holotype: HHUF 29192 Saprobic on vines of Actinidia deliciosa C. F. Liang & A. R. Ferguson. Sexual morph: Ascomata 519–548μm high× 257–310μm diam. (x = 330×280μm, n=10), solitary or gregarious, immersed to semi-immersed, coriaceous to carbonaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole slit-like, central, papillate, with a reduced crest,

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opening by an apical, lysigenous pore or dehiscence. Peridium 45–60μm wide, broad at the apex and thinner at the base, composed of two strata, outer stratum comprising several layers of brown to reddish brown, flattened cells of textura angularis fusing and indistinguishable from the host tissues and inner stratum comprising 2–3 layers of hyaline to lightly pigmented cells of textura intricata. Hamathecium comprising 1–1.5 μm wide, numerous, branch, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 101–140×8–13μm (x = 120×6μm, n=10), 8-spored, bitunicate, fissitunicate, cylindric or clavate, with a long pedicel, apically rounded, with an ocular chamber. Ascospores 14–16×6–8μm (x = 15.5× 6.9μm, n=15), uni seriate, partially overlapping, hyaline when immature, becoming brown when mature, broadly ellipsoid, 3-septate to muriform, with a single vertical septum, deeply constricted at the central septum, guttulate, smoothFig. 18 Platystomum crataegi (holotype). a. Appearance of ascomata on host surface b. Section through ascoma (note the crest-like papilla) c. Ostiole d. Peridium e. Pseudoparaphyses f– i. Asci j–n. Ascospores. Scale bars: b=100μm, c, d=20μm, e= 10μm, f–i=20μm, j–n=10μm


walled, lacking a mucilaginous sheath. Asexual morph: Undetermined. Material examined: JAPAN: Campus of Hirosaki University, Hirosaki, Aomori, on vines of Actinidia deliciosa (Actinidiaceae), 21 May 2001, K. Tanaka KT 521 (HHUF 29192, holotype), ex-type living culture KT 521; JAPAN: Ainai, Sauma, Aomori, on a submerged twigs of woody plant, 28 May 2001, K. Tanaka KT 534 (HHUF 29194), living culture JCM 13125; MAFF 239635. Notes: Platystomum actinidiae is introduced as a new species based on the material (HHUF 29192 and 29194) previously designated as of Platystomum compressum. Platystomum actinidiae can be distinguished from P. compressum based on its unique morphological characteristics and phylogenetic placement. Platystomum actinidiae differs from P. compressum in having a brown to reddish brown peridium with cells of textura


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Fig. 19 Platystomum crataegi (Asexual morph from the extype living culture). a Germinating ascospore b, c Colonies on PDA (c from below) d Conidiomata e Close up of conidioma. f Immature and mature conidia attached to conidiogenous cell. g Conidia. Scale bars: a, f, g =10μm

angularis to textura intricata and brown, 3-septate to muriform ascospores, deeply constricted at the central septum. In the present phylogenetic analysis, two strains of Platystomum actinidiae are clearly separated from the reference strain of P. compressum (MFLUCC 13-0343). Platystomum crataegi Wanasinghe, Thambugala, Camporesi & K.D. Hyde, sp. nov. Index Fungorum Number: IF551247; Facesoffungi number: FoF00817; Figs. 18 and 19 Etymology: Referring to the host genus Crataegus. Holotype: MFLU 15-1176 Saprobic on species of Crataegus in terrestrial habitats. Sexual morph: Ascomata 250–500μm high×275–300μm diam. (x = 351.1×288.1μm, n=5), scattered to gregarious, immersed, coriaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole crest-like, central, papillate, with a pore-like opening. Peridium 10–15μm wide at the

base, 10–20μm wide at the sides, broad at the apex, comprising two strata, outer stratum composed of small, brown to dark brown, small, somewhat flattened, thick-walled cells of textura angularis, fusing and indistinguishable from the host tissues, inner stratum composed of several layers with lightly pigmented to hyaline cells of textura angularis. Hamathecium comprising 1–2μm wide, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 80–120×15–17μm (x = 103.3×15.9μm, n=15), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicel, rounded at the apex, with an ocular chamber. Ascospores 22–28×9–11μm (x = 25.3×9.9μm, n=40), uni to bi-seriate, partially overlapping, initially hyaline, becoming brown at maturity, ellipsoidal to fusiform, muriform, 6–8 transversely septate, with 2–7 vertical septa, slightly curved, deeply constricted at the central septum, slightly constricted at the remaining septa, rounded at the ends, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Coelomycetous.

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Conidiomata 1.5–2 mm diam. pycnidial, phoma-like, solitary, gregarious, dark brown to black, slightly erumpent, papillate, with a sphaerical mass of slimy conidia oozing out at ostiolar apex. Conidiomata wall 170–250 μm wide, multi-layered, with 3–4 outer layers of brown-walled cells of textura angularis, with inner most layer thin, hyaline. Conidiophores short, septate, sparsely branched, formed from the inner most layer of pycnidium wall. Conidiogenous cells phialidic, cylindrical, hyaline, flexuous, smooth, with short collarette. Conidia 2.5–3.5×1.5–2μm (x = 3×1.7μm, n=50), hyaline, aseptate, straight to curved, ellipsoidal, with rounded ends, thin-walled, smooth-walled, numerous. Culture characteristics: Ascospores germinating on PDA within 24 h and producing germ tubes from both ends. Colonies on PDA growing slowly, reaching 30–35 mm diam. after 21 days at 16 °C, surface dirty white, spreading with moderate aerial mycelium, and even, smooth margins; reverse buff. Material examined: ITALY, Province of Arezzo [AR], Stia, Papiano, on dead and hanging branches of Crataegus sp. (Rosaceae), 9 April 2013, Erio Camporesi IT 1183 (MFLU Fig. 20 Platystomum rosae (holotype). a. Appearance of ascomata on host surface b. Section through ascoma c. Ostiole d. Peridium e. Pseudoparaphyses f, g. Immature and mature asci h– k. Ascospores. Scale bars: b= 100μm, c=20μm, d, e=10μm, f,g=20μm, h-k=10μm


15-1176, holotype);. ibid., (HKAS, isotype), ex-type living culture, MFLUCC 14-0925, KUMCC. Notes: Platystomum crataegi is introduced here as a new species based on its morphological traits and phylogenetic placement. Platystomum crataegi differs from other Platystomum species in having lightly pigmented, muriform, 6–8 transversely septate, ascospores with 2–7 vertical septa. Although this species has some similar morphological characters with Pleospora species, the crest-like, papillate ostiole, and ellipsoidal ascospores with a deeply constricted central septum and lack of a mucilaginous sheath differ from Pleospora species. Platystomum rosae Wanasinghe, Thambugala, Camporesi, & K.D. Hyde, sp. nov. Index Fungorum Number: IF551248; Facesoffungi number: FoF00818; Fig. 20 Etymology: Referring to the host genus Rosa. Holotype: MFLU 15-1177 Saprobic on species of Rosa, in terrestrial environments. Sexual morph: Ascomata 300–410μm high×250–400μm


diam. (x = 366.8×337.9μm, n=5), scattered to gregarious, immersed, coriaceous, dark brown to black, subglobose to conical, ostiolate. Ostiole slit-like, central, papillate, opening by an apical, lysigenous pore or dehiscence, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 15–25μm wide at the base, 18– 30μm wide at the sides, broad at the apex, comprising two strata, outer stratum comprising dark brown, somewhat flattened cells of textura angularis, fusing and indistinguishable from the host tissues, and inner layers with several layers of lightly pigmented to hyaline cells of textura angularis. Hamathecium comprising 1.5–2.5μm wide, unbranched, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 110–150×13–15μm (x = 130.6×13.9μm, n=15), 8-spored, bitunicate, fissitunicate, cylindrical to cylindric-clavate, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores 20–25×7– 9μm (x = 22.2×8.1μm, n=40), uni to bi-seriate, partially overlapping, initially hyaline, becoming brown at maturity, ellipsoidal to fusiform, muriform, 3–6 transversely septate, with 1–4 vertical septa, slightly curved, deeply constricted at the central septum, broadly to narrowly rounded at the ends, guttulate, lacking a mucilaginous sheath. Asexual morph: Undetermined. Culture characteristics: Ascospores germinating on PDA within 24 h and producing germ tubes from both ends. Colonies on PDA growing slowly, reaching 30–35 mm diam. after 21 days at 16 °C, greenish-grey in centre, with dirty white in outer region, spreading with moderate aerial mycelium, and even, smooth margins; reverse, buff-yellow. Material examined: ITALY, Province of Forlì-Cesena [FC], Fiumicello di Premilcuore, on dead and hanging branches of Rosa canina L. (Rosaceae), 20 January 2014, Erio Camporesi IT 1660 (MFLU 15-1177, holotype);. ibid., (HKAS, isotype), ex-type living culture, MFLUCC 15–0633, KUMCC. Notes: Platystomum rosae is morphologically very similar to P. salicicola, but differs in having small, subglobose to conical ascomata, an ostiole without periphyses, a thin peridium and light brown, 3–6 transversely septate ascospores, with 1–4 vertical septa. Platystomum salicicola Wanasinghe, Thambugala, Camporesi, & K.D. Hyde, sp. nov. Index Fungorum Number: IF551249; Facesoffungi number: FoF 00819; Fig. 21 Etymology: Referring to the host genus Salix. Holotype: MFLU 15-0201 Saprobic on species of Salix, in terrestrial habitats. Sexual morph: Ascomata 300–450μm high×250–550μm diam. (x = 385×396μm, n=5), immersed, coriaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole slit-like, central, papillate, periphysate, with a crest-like apex and opening by an apical, lysigenous pore or dehiscence. Peridium 15– 25μm wide at the base, 25–35μm wide at the sides, broad at

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the apex, comprising two strata, outer stratum comprising dark brown, somewhat flattened cells of textura prismatica to textura angularis, fusing and indistinguishable from the host tissues, and inner layers comprising several layers of lightly pigmented to hyaline cells of textura angularis. Hamathecium comprising 1–2.5 μm wide, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 110–140×14–17μm (x = 127.2×15.4μm, n=15), 8-spored, bitunicate, fissitunicate, cylindrical to cylindric-clavate, with a long pedicel, rounded at the apex, with an ocular chamber. Ascospores 20–25×7– 10μm (x = 22.3×8.3μm, n=40), uni to bi-seriate, partially overlapping initially hyaline, becoming brown to dark brown at maturity, ellipsoidal to fusiform, muriform, 4–6 transversely septate, with 2–4 vertical septa, deeply constricted at the central septum, slightly constricted at the remaining septa, broadly to narrowly rounded at the ends, guttulate, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined. Culture characteristics: Ascospores germinating on PDA within 24 h and producing germ tubes from both ends. Colonies on PDA growing slowly, reaching 25–30 mm diam. after 21 days at 16 °C, surface dirty white, spreading with moderate aerial mycelium, and even, smooth margins; reverse buff. Material examined: ITALY, Province of Forlì-Cesena [FC], Fiumicello di Premilcuore, on dead and hanging branches of Salix sp. (Salicaceae), 8 Dec 2013, Erio Camporesi IT 1567 (MFLU 15-0201, holotype);. ibid., (HKAS, isotype), ex-type living culture, MFLUCC 15-0632, KUMCC. Pseudolophiostoma Thambugala, Kaz. Tanaka & K.D. Hyde, gen. nov. Index Fungorum number: IF551250; Facesoffungi number: FoF00820 Etymology: Referring to its similarity with Lophiostoma. Saprobic on dead twigs. Sexual morph: Ascomata scattered, immersed, coriaceous to carbonaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole crest-like, central, broadly papillate, with an irregular pore-like opening, plugged by hyaline, filamentous hyphae, and occasionally lighter coloured. Peridium composed of a single stratum, with 4–6 layers of dark brown to black cells of textura prismatica, fusing and indistinguishable from the host tissues. Hamathecium comprising 1–2.5μm wide, numerous, filamentous, septate, branched, anastomosing pseudoparaphyses, situated between and above the asci embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a short pedicel, apically rounded, with an ocular chamber. Ascospores uni to bi-seriate, hyaline, fusiform, 1septate, with 2 euseptate, constricted at the septum, with a narrow sheath. Asexual morph: Undetermined. Type species: Pseudolophiostoma vitigenum (Kaz. Tanaka & Y. Harada) Thambugala, Kaz. Tanaka & K.D. Hyde, comb. nov.

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Fig. 21 Platystomum salicicola (holotype). a. Appearance of ascomata on host surface b. Section through ascoma. note the crest-like ostiole c. Pseudoparaphyses d. Peridium e, f. Asci g–l. Ascospores. Scale bars: b=100μm, c=10μm, d–f= 20μm, g–l=10μm

Basionym: Lophiotrema vitigenum Kaz. Tanaka & Y. Harada, Mycoscience 44(2): 119 (2003) ≡ Lophiostoma vitigenum (Kaz. Tanaka & Y. Harada) K. Hiray. & Kaz. Tanaka, Mycoscience 52(6): 407 (2011) Index Fungorum number: IF551252; Facesoffungi number: FoF008821; Fig. 22 Saprobic on dead vines of Vitis coignetiae Pulliat ex Planch. Sexual morph: Ascomata 450–570μm high, 350–

410μm diam., scattered, immersed, papilla erumpent through host surface, coriaceous to carbonaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole crest-like, central, periphysate, broadly papillate, with an irregular pore-like opening, plugged by hyaline, filamentous hyphae, and occasionally lighter coloured. Peridium 13–25μm wide, composed of a single stratum, with 4–6 layers of dark brown to black cells of textura prismatica, fusing and


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Fig. 22 Pseudolophiostoma vitigenum (holotype). a, b. Appearance of ascomata on host surface with crest-like papilla c, d. Ascomata in longitudinal section (in lactophenol cotton blue) e. Peridium f.

Pseudoparaphyses g, h. Asci i–l. Ascospores (j. in black-blue ink) m. Germinating ascospore. Scale bars. a, b=200μm, c, d=50μm, e–h, l, m=10μm, i–k=5μm

indistinguishable from the host tissues. Hamathecium comprising 1–2.5μm wide, numerous, filamentous, septate, branched, anastomosing pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci (105–)115–147(–158)×(17.5–)18.5–22 (–23) μm (x = 129.8 × 20.1 μm, n = 50), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a short pedicel, apically rounded, with an ocular chamber. Ascospores (30.5–)34–44(–51) × (8–)9–11.5(–13) μm (x = 38.5 × 10.5μm, n=141), L/W (3–)3.2–4.3(–5) (x = 3.7, n=141), uni to bi-seriate, hyaline, fusiform, 1-septate, with 2 euseptate, constricted at the septum, guttulate, smoothwalled, with a narrow sheath. Sheath drawn out at the ends, 3–12 μm long at both ends. Asexual morph: Undetermined. Notes: Lophiostoma vitigenum Kaz. Tanaka & Y. Harada was introduced as Lophiotrema vitigenum Kaz. Tanaka & Y.

Harada based on their rather small ascomata and peridia of equal thickness (Tanaka and Harada 2003b). The phylogenetic analysis and morphological re-examination of L. vitigenum revealed that this species belongs to Lophiostoma (Hirayama and Tanaka 2011). In the present phylogeny (Fig. 1) the extype strain of L. vitigenum (MAFF 239459) clustered in subclade A13 (Fig. 1). We introduce a new genus Pseudolophiostoma to accommodate Lophiostoma vitigenum in Lophiostomataceae. Material examined: JAPAN, Aomori, Hirosaki, Kudoji, on vines of Vitis coignetiae. (Vitaceae), 27 October 2001, S. Hatakeyama (HHUF 26930 holotype); ex-type living culture JCM 13534, MAFF 239459; ibid., (HHUF 26931, isotype), ex-isotype living culture JCM 17676. Pseudoplatystomum Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551253; Facesoffungi number: FoF00822

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Etymology: Referring to its similarity with Platystomum. Saprobic on marine driftwood. Sexual morph: Ascomata solitary or gregarious, immersed to slightly erumpent, subglobose, black, coriaceous, ostiolate. Ostiole slit-like, central, rounded, broadly papillate, ostiolar canal cylindrical, filled with a network of branched, septate periphyses. Peridium composed of two strata, outer layer wide at the base and the sides, wider near the ostiole, composed of melanized cells forming textura angularis, fusing with the host tissue, and the inner stratum thick at the base, composed of hyaline, rounded or weakly angular small cells that merge with the pseudoparaphyses. Hamathecium comprising 1.2–2μm wide, trabeculae pseudoparaphyses, unbranched at the base, becoming branched and anastomosing above the asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical, pedicellate, apically rounded, with an ocular chamber. Ascospores uni seriate, brown, fusiform, muriform, with 5–7(−8) transverse septa and 1–2(−3) vertical septa, constricted at the central septum, rough-walled or verrucose, lacking a mucilaginous sheath (Abdel-Wahab and Jones (2000). Asexual morph: Undetermined. Type species: Pseudoplatystomum scabridisporum (Abdel-Wahab & E.B.G. Jones) Thambugala & K.D. Hyde, comb. nov. Basionym: Platystomum scabridisporum Abdel-Wahab & E.B.G. Jones, Mycoscience 41(4): 384 (2000) Index Fungorum Number: IF551254 Notes: Platystomum scabridisporum was introduced by Abdel-Wahab and Jones (2000) in Platystomaceae based on morphological characteristics. Suetrong et al. (2009) showed that two strains of P. scabridisporum (BCC 22835 and BCC 22836) clustered in Lophiostomataceae in their phylogenetic analysis. In our analysis, the placement of P. scabridisporum is confirmed in Lophiostomataceae (clade A11, Fig. 1). Suetrong et al. ( 2009) did not formally transfer P. s c a b r i d i s p o r u m t o L o p h i o s t o m a t a c e a e . Pseudoplatystomum scabridisporum differs from Platystomum compressum in having smaller ascomata, broadly papillate ostiole with periphyses, wider peridium with cells of textura angularis, longer asci and the greater number of transverse and vertical septa in ascospores with verrucose walls (Abdel-Wahab and Jones 2000). Sigarispora Thambugala & K.D. Hyde, gen. nov. Index Fungorum Number: IF551255; Facesoffungi number: FoF00823 Etymology: Referring to the cigar-shaped ascospores in the type species. Saprobic on decaying culms of grasses (Poaceae) or dead herbaceous stems. Sexual morph: Ascomata solitary, scattered to gregarious, immersed, papilla erumpent through host surface, black, globose to subglobose, ostiolate. Ostiole crest or slit-like, central, rounded, papillate, with a pore-like opening, plugged by gelatinous tissue, made up of lightly


pigmented, pseudoparenchymatous cells. Peridium composed small, light brown to dark brown, thin-walled cells of textura angularis, fusing at the outside with the host tissues. Hamathecium comprising 1–3μm wide, numerous, long, cellular, septate, branched, pseudoparaphyses, embedded in a gelatinous matrix, between and above the asci. Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, long pedicellate, apically rounded, with an ocular chamber. Ascospores uni to bi-seriate, overlapping, yellowish brown to dark brown, ellipsoidal-fusiform or cigar-shaped, 5–6 transversely septate or muriform, with 2–4 vertical septa, constricted at the central septum, smooth-walled, with or without a mucilaginous sheath, sometimes with terminal appendages. Asexual morph: Undetermined. Type species: Sigarispora ravennica (Tibpromma, Camporesi & K.D. Hyde) Thambugala & K.D. Hyde, comb. nov. Basionym: Lophiostoma ravennicum Tibpromma et al., in Liu et al., Fungal Diversity 72(1): 117 (2015) Index Fungorum Number: IF551256 Notes: Lophiostoma ravennicum was introduced in Liu et al. (2015) based on its phylogenetic placement in Lophiostomataceae. Its cigar-shaped, usually 6-septate, brown ascospores, with wide mucilaginous sheath are unlike Lophiostoma sensu stricto, which has hyaline ascospores. Lophiostoma ravennicum grouped in clade A5 in the phylogenetic analyses (Fig. 1) with new strain (MFLUCC 14-0941) and putatively named Lophiostoma strains L. arundinis, L. caudatum, and L. caulium Bvar. a^. We transfer these species to Sigarispora based on their morohological similarities and phylogenetic placement. The distinct characters of Sigarispora include immersed to semi-immersed ascomata, a small crest-like ostiole, a peridium of elongate cells of textura angularis, and brown cigar-shaped, multi-septate ascospores. Sigarispora coronillae (MFLUCC 14-0941) differs from S. ravennica in having 3–5 transversely septate, muriform ascospores without a mucilaginous sheath. Sigarispora caulium (Fr.) Thambugala, Wanasinghe, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Lophiostoma caulium (Fr.) Ces. & De Not., Commun. Soc. Critt. Ital. 1:219, 1863 Index Fungorum Number: IF551532; Facesoffungi number: FoF00824; Fig. 23 Saprobic on dead herbaceous twigs. Sexual morph: Ascomata 180–340μm high, 200–280μm diam, solitary, immersed, coriaceous, black, subglobose, ostiolate. Ostiole rounded or slit-like, variable in shape, up to 40–120μm long and 20–40μm wide, central, papillate, with a crest like apex and a pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 10–20μm wide (x = 16.25μm, n=20), wider at the apex, thinner at the base, comprising a single stratum, with lightly pigmented, thick-walled cells of textura angularis,


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Fig. 23 Sigarispora caulium (HHUF 27306). a, b. Herbarium material. c. Appearance of ascomata on host surface. d. Section through of ascoma e. Ostiole. f. Peridium. g–i. Asci. j. Pseudoparaphyses. k–n. Ascospores. Scale bars: d=50μm, e=20μm, f–i=10μm, j–n=5μm

cells towards the inside lighter, at the outside, darker, somewhat flattened, fusing and indistinguishable from the host tissues. Hamathecium comprising 2–3μm wide, numerous, filamentous, branched, septate, pseudoparaphyses, situated between and above the asci embedded in a gelatinous matrix. Asci 75–100×12–14μm (x = 84.6×13.1μm, n=40), 8-spored,

bitunicate, fissitunicate, cylindric-clavate, with a short, bulbous pedicel, rounded at the apex, with an ocular chamber. Ascospores 18–25×5–8μm (x = 22.6×6.1μm, n=50), uni to bi-seriate, partially overlapping, hyaline when young, becoming yellowish brown at maturity, fusiform with narrow, acute ends, mostly curved, 5-septate, constricted at the central

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septum, cells above central septum swollen, guttulate, smooth-walled, with appendages at both ends. Asexual morph: Undetermined. Material examined: JAPAN, Oowasawa River, river bank, Kadoke, Hirosaki, Aomori, on stems of unknown herbaceous plant, 29 July 2001, Kazuaki Tanaka KT. 603 (HHUF 27306), living culture, MAFF 239450; JAPAN, Sanpinai, Hirosaki, Aomori, on stems of unknown herbaceous plant, 29 July 2001, Kazuaki Tanaka KT. 633 (HHUF 27307), living culture JCM 17669. Fig. 24 Sigarispora arundinis (HHUF 27305). a, b. Herbarium specimen c, d. Appearance of ascomata on host surface e. Sections through ascoma f. Ostiole g. Sections throught peridium h. Pseudoparaphyses i– l. Asci m–q. Ascospores (q from HHUF 27304) . Scale bars: b, c= 500μm, d=50μm, f, g=25μm, i– l=10μm, h, m–q=5μm


Notes: Sigarispora caulium is introduced here to accommodate Lophiostoma caulium (Fr.) Ces. & De Not. Three reference strains of Lophiostoma caulium (MFLUCC 150176, MAFF 239450 and JCM 17669) clustered in the Sigarispora clade and Lophiostoma caulium shares similar morphological characteristics with Sigarispora such as immersed ascomata with a crest-like ostiole, cylindric-clavate asci and brown, ellipsoidal-fusiform, multiseptate ascospores, while mainly differs from other Sigarispora species in having ascospores with appendages at both ends.


Sigarispora arundinis (Pers.) Thambugala, Qing Tian, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Sphaeria cristata ß arundinis Pers., Syn. meth. fung. (Göttingen) 1: 54 (1801) ≡ Lophiostoma arundinis (Pers.) Ces. & De Not., Schem. di Classif. Sferiacei: 46 (1863) Index Fungorum Number: IF551533; Facesoffungi number: FoF00825; Fig. 24 Saprobic on culm of Phragmites australis (Cav.) Trin. Ex Steud. in terrestrial habitats. Sexual morph: Ascomata 320– 416μm high×250–310μm diam. (x = 383×276μm, n=10), scattered to gregarious, immersed to semi-immersed, papilla erumpent through host surface, coriaceous, dark brown to black, ostiolate. Ostiole slit-like, variable in shape, up to 120 μm long and 249 μm wide, central, papillate, with a crest-like apex and a pore-like opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 32–53μm wide (x = 49μm, n=10), wider at the apex and thinner at the base, comprising two strata, outer stratum composed of dark brown to black, flattened, thickwalled cells of textura angularis, fusing and indistinguishable from the host tissues, inner stratum composed of several layers with lightly pigmented to hyaline, somewhat fattened cells of textura angularis. Hamathecium comprising 1–2μm wide, septate, cellular pseudoparaphyses, branching and anastomosing between and above the asci, embedded in a gelatinous matrix. Asci 94–112.5×12.5–14.5μm (x = 104.5×13.8μm, n=10), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a short, bulbous pedicel, apically rounded with an ocular chamber. Ascospores 22–32.5×6.3–7.9μm (x = 25.5×7.2μm, n = 10), uni to bi-seriate, partially overlapping, lightly pigmented to brown, fusiform with narrow, acute ends, usually 5-septate at maturity, constricted at the central septum, guttulate, smooth-walled. Asexual morph: Undetermined. Material examined: JAPAN: Hirakawa, river bank, Hiraka, Aomori, on culm of Phragmites australis (Poaceae), 5 August 2001, Kazuaki Tanaka KT. 651 (HHUF 27305, reference specimen designated here); living culture, MAFF 239449, JCM 13551; JAPAN: Oowasawa River, river bank, Kadoke, Hirosaki, Aomori, 29 July 2001, On culms of Phragmites australis (Poaceae), Kazuaki Tanaka KT 606 (HHUF 27304), living culture JCM 13550. Notes: Lophiostoma arundinis (Pers.) Ces. & De Not. shares similar morphological traits with Sigarispora and putatively named strains of L. arundinis (JCM 13551 and JCM 13550) grouped in the Sigarispora clade with high support. We observed and illustrated the reference specimens (HHUF 27305 and HHUF 27304) attached to these strains and they show typical characteristics of L. arundinis (Chesters and Bell 1970, Holm and Holm 1988). Therefore we introduce Lophiostoma arundinis as a new combination in Sigarispora. Sigarispora arundinis differs from other Sigarispora species in having wide peridium consists with cells of textura

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angularis to textura prismatica and initially 1-septate ascospores becoming 5-septate at maturity. Sigarispora caudata (Fabre) Thambugala, Qing Tian, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Lophiostoma caudatum Fabre, Annls Sci. Nat., Bot., sér. 6 9: 103 (1879) [1878] Index Fungorum Number: IF551534; Facesoffungi number: FoF00826; Fig. 25 Saprobic on culms of Dactylis glomerata L. in terrestrial habitats. Sexual morph: Ascomata 145–210μm high×210– 305μm diam. (x = 173×251μm, n=10), scattered to gregarious, immersed to semi-immersed, papilla erumpent through host surface, flattened at the base, coriaceous, black, usually staining the substrate around the ascomata with black shade, ostiolate. Ostiole slit-like, variable in shape, up to 180μm long and 115μm wide, central, with a crest-like apex and a porelike opening, plugged by gelatinous tissue, made up of lightly pigmented, pseudoparenchymatous cells. Peridium 18–32μm wide (x = 27μm, n=12), wider at the apex and thinner at the base, comprising a single stratum, with brown to lightly pigmented, thin-walled cells of textura angularis, cells towards the inside lighter, at the outside, darker, somewhat flattened, fusing and with the host tissues. Hamathecium comprising 1–2μm wide, septate, cellular pseudoparaphyses, branching and anastomosing between and above the asci, embedded in a gelatinous matrix. Asci 86–112.5×10.5–13μm (x = 105.5×12.5μm, n=10), 8-spored, bitunicate, fissitunicate, clavate, with a long, bulbous pedicel, apically rounded with a distinct ocular chamber. Ascospores 23.5–34.5×5.5–7μm (x = 26.5×6.2μm, n=10), bi-seriate, overlapping, brown to dark brown, fusiform with narrow, acute ends, usually (4)–5–(6)septate, constricted at the central septum, guttulate, smoothwalled, lacking a mucilaginous sheath. Asexual morph: Undetermined. Material examined: JAPAN: Hirosaki University, Hirosaki, Aomori, on culm of Dactylis glomerata (Poaceae), 21 May 2001, Kazuaki Tanaka KT 530 (HHUF 27319), living culture, MAFF 239453. Notes: Sigarispora caudatum is introduced here based on Lophiostoma caudatum Fabre and the shares similar morphological characteristics with Sigarispora such as semiimmersed ascomata with a crest-like spex, and brown, multiseptate fusiform ascospores, while differs from other Sigarispora species in having ascomata with a flattened base and well-developed crest-like apex. Sigarispora coronillae Wanasinghe, Thambugala, Camporesi, & K.D. Hyde, sp. nov. Index Fungorum Number: IF551257; Facesoffungi number: FoF00827; Fig. 26 Etymology: Referring to the host genus Coronilla. Holotype: MFLU 15–1178 Saprobic on species of Coronilla, in terrestrial habitats. Sexual morph: Ascomata 350–400μm high×390–450μm

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diam. (x = 390.2×413.1μm, n=5), scattered to gregarious, immersed, coriaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole crest-like, central, rounded, papillate, periphysate, a pore-like opening. Peridium 20–40μm wide at the base, wider at the sides, composed of two strata, outer stratum comprising dark brown to black, flattened, thick-walled cells of textura angularis, fusing and indistinguishable from the host tissues, inner stratum composed of several layers of brown to lightly pigmented cells of textura angularis. Hamathecium comprising 1.5–2.5μm wide, septate, hyaline, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 120–140×14–17μm (x = 129.3×15.4μm, n=15), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a long pedicell, rounded at the apex, with an ocular chamber. Ascospores 20–26×8–10μm (x = 23.4×9.1μm, n=40), uni to bi-seriate, overlapping, initially hyaline, becoming brown at maturity, ellipsoid to fusiform, broadly to narrowly rounded at the ends, muriform, with 4–5 transverse septa, and 2–4 vertical septa, slightly curved, constricted at the central septum, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined. Culture characteristics: Ascospores germinating on PDA within 24 h and producing germ tubes from both ends. Colonies on PDA growing slowly, reaching 30–35 mm diam. after 21 days at 16 °C, pale olivaceous-grey in outer region, spreading with moderate aerial mycelium, and even, smooth margins; reverse; olivaceous-grey. Material examined: ITALY, Province of Forlì-Cesena [FC], San Piero in Bagno, Valgianna, on dead and hanging branches of Coronilla emerus L. (Fabaceae), 6 February 2014, Erio Camporesi IT 1704 (MFLU 15-1178, holotype);. ibid., (HKAS, isotype), ex-type living culture, MFLUCC 140941, KUMCC. Notes: Sigarispora coronillae is introduced here as a new species from Italy and morphologically clearly differs from other Sigarispora species in having muriform ascospores. The ex-type strain of Sigarispora coronillae (MFLUCC 140941) clustered in the Sigarispora clade together with Sigarispora ravennica and Sigarispora caudatum. Vaginatispora K.D. Hyde, Nova Hedwigia 61(1–2): 234 (1995) Facesoffungi number: FoF00828 Saprobic on submerged wood in fresh water or herbaceous plants in terrestrial habitats. Asexual morph: Ascomata scattered to gregarious, semi-immersed to erumpent, coriaceous, black, subglobose, ostiolate. Ostiole slot-like, central, periphysate or not, papillate, with a pore-like opening. Peridium wider at the apex and thinner at the base, composed of hyaline to brown textura angularis, cells towards the inside lighter, at the outside, darker, fusing at the outside with the host cells. Hamathecium of 1.5–2.5μm wide, numerous, hyaline, septate, cellular pseudoparaphyses, situated between and

Fungal Diversity (2015) 74:199–266 Fig. 25 Sigarispora caudata (holotype). a, b. Herbarium specimens c, d. Appearance of ascomata on host surface. e, f. Sections through ascoma. g. Ostiole. h. Sections through peridium. i–l. Asci. m. Pseudoparaphyses. n–p. Ascospores. Scale bars: c=200μm, d=100μm, e, f=50μm, g, h= 25μm, i–l=10μm, h, m-p=5μm

above the asci, embedded in a gelatinous matrix. Asci 8spored, fissitunicate, cylindrical-clavate, with a pedicel, apically round with an ocular chamber. Ascospores mostly 2seriate, fusiform with acute ends, 1-septate, sometimes euseptate, constricted at the septum, upper cell slightly broader than lower cell, hyaline, with or lacking a mucilaginous sheath or terminal appendages. Asexual morph: Undetermined. Type species: Vaginatispora aquatica K.D. Hyde, Nova Hedwigia 61(1–2): 235 (1995) ≡ Lophiostoma vaginatispora Zhang et al., Phytotaxa 176(1): 177 (2014) Notes: Vaginatispora was introduced by Hyde (1995) in Massarinaceae to accommodate Vaginatispora aquatica and is characterised by immersed to erumpent ascomata, with slotlike ostioles, with numerous periphyses, and 1-septate, narrowly fusiform ascospores, with a thick surrounding papilionaceous sheath. Zhang et al. (2014) showed that V. aquatica had phylogenetic affinities with Lophiostomataceae based on new collection from Thailand and transferred the species to Lophiostoma. In the present phylogenetic analysis L. vaginatispora (MFLUCC 11-0083) nested in clade B (subclade B1, Fig. 1) along with the putative strains of L. fuckelii (MAFF 239458, JCM 17672) and L. alpigenum Fuckel. In this study we accept Vaginatispora as a separate genus in Lophiostomataceae and based on morphological characteristics and phylogeny, Lophiostoma fuckelii is transferred to the genus Vaginatispora. Vaginatispora fuckelii differs from V. aquatica in having erumpent ascomata, a paridium composed of two strata and ascospores with appendages at both ends. Vaginatispora fuckelii (Sacc.) Thambugala, Wanasinghe, Kaz. Tanaka & K.D. Hyde, comb. nov. Basionym: Lophiostoma fuckelii Sacc., Michelia 1(no. 3): 336 (1878) Index Fungorum Number: IF551535; Facesoffungi number: FoF00829; Fig. 27 Saprobic on dead herbaceous twigs. Sexual morph: Ascomata 150–180μm high, 200–250 μm diam, solitary, semi-immersed to erumpent, coriaceous, black, subglobose, ostiolate. Ostiole rounded or slit-like, variable in shape, up to 50–75μm long and 30–40μm wide, central, with a porelike opening. Peridium 15–25μm wide (x = 21.7μm, n=20), wider at the apex and thinner at the base, composed of two strata, outer stratum comprising light brown to brown, somewhat flattened cells of textura angularis, fusing at the outside with the host cells, and inner stratum comprising several


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Fig. 26 Sigarispora coronillae (holotype). a. Appearance of ascomata host surface b. Section through ascoma c. Ostiole d. Peridium e. Pseudoparaphyses f, i. Asci j–n. Ascospores. Scale bars: b=100μm, c=20μm, d, e= 10μm, f–i=20μm, j–n=10μm

layers of hyaline cells of textura angularis. Hamathecium comprising 1.5–2.5μm wide, numerous, branched, septate, cellular pseudoparaphyses, situated between and above the asci, embedded in a gelatinous matrix. Asci 60–90×9–12μm (x = 75.7×10.4μm, n=40), 8-spored, bitunicate, fissitunicate, cylindric-clavate, with a short, bulbous pedicel, rounded at the apex, with an indistinct ocular chamber. Ascospores 15–17× 4–5μm (x = 15.5×4.3μm, n=50), uniseriate to bi-seriate, fusiform with acute ends, 1-septate, 2-euseptate, constricted at the septum, upper cell slightly broader than lower cell, hyaline, guttulate, smooth-walled, with globose appendages at both ends. Asexual morph: Undetermined. Material examined: JAPAN, Sanpinai, Hirosaki, Aomori, on stems of unknown herbaceous plant, 5 August 2001, Kazuaki Tanaka KT 634 (HHUF 27325), living culture, MAFF 239458. JAPAN, Shinrin Park, Hokkaido, on Vitis coignetiae (Vitaceae), 25 July 2008, Kazuaki Hirayama KH 161 (HHUF 30076), living culture, JCM 17672.

Floricolaceae Thambugala, Kaz. Tanaka & K. D. Hyde, fam. nov. Index Fungorum number: IF551258; Facesoffungi number: FoF00830 Saprobic on twigs, branches or bark of various woody and herbaceous plants in terrestrial habitats. Sexual morph: Ascomata solitary, or scattered to gregarious, immersed to erumpent or superficial, dark brown to black, pyriform, globose, or globose or subglobose, with or without a subiculum, coriaceous to carbonaceous, ostiolate. Ostiole central, area light coloured or not and ostiole opening appearing plugged by gelatinous tissue, ostiolar canal filled with periphyses, with a pore-like opening. Peridium thick at the apex, thinner at the base, comprising two cell types, outer stratum composed of small heavily pigmented, dark brown, thick-walled, compressed cells of textura angularis, inner stratum thin, composed of smaller, hyaline cells.


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Fig. 27 Vaginatispora fuckelii (HHUF 27325). a, b. Herbarium labels. c. Appearance of ascomata on host surface. d. Section through ascoma e. Section through peridium. f. Pseudoparaphyses. g, h. Asci. i– k. Ascospores (i in Black-blue ink). Scale bars: d=50μm, e= 10μm, f=5μm, g–k=10μm

Hamathecium comprising 1–2μm wide, numerous, septate, branched, cellular or filamentous pseudoparaphyses, anastomosing above the asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate cylindroclavate, pedicellate, rounded at the apex, with an ocular chamber. Ascospores uni to bi-seriate, hyaline to dark brown, fusiform, ovoid, or oblong to elliptical, 1 to multi-septate or muriform, and constricted at the central septum, often guttulate, smooth-walled, with or without a mucilaginous sheath. Asexual morph: coelomycetous. Conidiomata pycnidial, subepidermal, substromatal, separate, dark brown, immersed, separate unilocular, globose or subglobose to depressed ellipsoidal, ostiolate. Ostiole central, papillate or not. Conidiomata wall

composed of several-layered of textura angularis; outer layers thick-walled, brown becoming thinner-walled and hyaline cells of in the inner layers. Conidiophores absent or 0–2-septate and branched at the base. Conidiogenous cells with or without annellations, discrete, smooth, indeterminate, cylindrical, hyaline to pale brown, branched, conidiogenesis holoblastic, proliferating 1–3 times precurrently, arising from the inner cells of the pycnidial wall. Conidia cylindrical to elongate ellipsoidal or ellipsoidal to oblong, aseptate or 3distoseptate, pale to dark brown, smooth-walled to verruculose, with truncate base and apex obtuse or rounded, sometimes surrounded by a thin mucilaginous sheath (Asexual morph follows Floricola striata in

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Kohlmeyer and Volkmann-Kohlmeyer (2000) and Coniothyrium grandicipis in Crous et al. (2004). Key to genera of Floricolaceae 1. Asexual morph with 1–3-septate, dark brown conidia with rounded apex and truncate base, sexual morph unknown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Floricola 1. Asexual morph not as above, sexual morph known . . 2 2. Ascomata immersed. . . . . . . . . . . . . . . Ramusculicola 2. Ascomata semi-immersed t o e rumpent, or superficial . . . . . . . . . . . . . . . . . . . . . . . . . . 3 3. Ascomata with a long neck . . . . . . . . . . . .Neocurreya 3. Ascomata without a long neck . . . . . . . . . . . . . . . . . . 4 4. Ascospores hyaline . . . . . . . . . . . . . . . . . . . . . . . . . . 5 4. Ascospores pigmented, brown to dark brown . . . . . . . 6 5. Ostiolar area orange . . . . . . . . . . . . . . . Aurantiascoma 5. Ostiolar area lighter, not orange Pseudoaurantiascoma 6. Ascomata solitary, or aggregated in large clusters on the host surface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 6. Ascomata solitary, or aggregated in small clusters on the host surface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 7. Ascomata superficial, with or without a subiculum. . . . . . . . . . . . . . . . . . . . . . . . Misturatosphaeria 7. Ascomata erumpent, lacking a subiculum. . . . . . . . . . . . . . . . . . . . . . . .Magnibotryascoma 8. Ascospores asymmetrical, fusiform to broadly clavate, with a somewhat tapering lower end. . . . . . Asymmetrispora 8. Ascospores mostly symmetrical, oblong to elliptical, with a rounded lower end . . . . . . . Pseudomisturatosphaeria Type: Floricola Kohlm. & Volkm.-Kohlm., Bot. Mar. 43(4): 385 (2000) Saprobic on dead leaves, inflorescences or branches of Juncus roemenanus and Vitis vinifera. Sexual morph: Undetermined. Asexual morph: Conidiomata pycnidial, scattered, immersed to erumpent, unilocular, subglobose to depressed ellipsoidal, ostiolate. Ostiole central, rounded, shortly papillate, light to dark brown, ostiolar canal lined with papillate and filled with a gelatinous mass. Conidiomata wall composed of 5–6 layers of polygonal, thicked walled, hyaline to brown cells of textura angularis merging with host tissues externally. C o n i d i o p h o re s r e d u c e d t o c o n i d i o g e n o u s c e l l s . Conidiogenous cells with annellations, enteroblastic, discrete, phialidic, determinate, doliiform or conical to cylindrical, hyaline. Conidia cylindrical to elongate ellipsoidal, with rounded apex and with truncate base with an abscission scar, 3distoseptate or with 1–3-longitudinal septa, constricted at some septa, dark brown, smooth-walled to verruculose, surrounded by a thin mucilaginous sheath or not (Kohlmeyer and Volkmann-Kohlmeyer 2000, Ariyawansa et al. 2015). Notes: The genus Floricola was established to accommodate the coelomycetous taxon F. striata. (Kohlmeyer and Volkmann-Kohlmeyer 2000). Ariyawansa et al. (2015)


introduced F. viticola. Two strains of F. striata (JK 56781 and JK 5603K), including the ex-type strain and the ex-type strain of Floricola viticola clustered with Misturatosphaeria in clade C (Fig. 1). No asexual morph has been reported for Misturatosphaeria (Mugambi and Huhndorf 2009) and no sexual morph is linked for Floricola (Volkmann-Kohlmeyer 2000). Curreya grandicipis, which has a coniothyrium-like asexual morph (Crous et al. 2011), also clustered in the Floricolaceae clade. Morphologically we cannot confirm the relationship of Floricola with Misturatosphaeria. Therefore, we maintain Floricola and Misturatosphaeria as separate genera and accommodate them in a new family Floricolaceae. The name of the new family Floricolaceae is selected based on the older genus Floricola. Phylogenetically, they form a well-supported clade distinctly separate from Lophiostomataceae. Misturatosphaeria species are polyphyletic and morphologically distinct. Therefore, we introduce six new genera to accommodate these distinct taxa. Here we introduce Aurantiascoma for M. minima and Pseudoaurantiascoma for M. kenyensis. The new genera Magnibotryascoma and Pseudomisturatosphaeria represent M . u n i s e r i a t a a n d M . c r u c i f o r m i s r e s p e c t i v e l y. Misturatosphaeria tennesseensis and Misturatosphaeria mariae are accommodated in Asymmetrispora. A new genus Ramusculicola is introduced for a new collection from Thailand. Curreya austroafricana, C. grandicipis and C. proteae clustered in clade C7 along with Misturatosphaeria uniseriata. The asexual morphs of three Curreya species have been reported as coniothyrium-like (Crous et al. 2004; Marincowitz et al. 2008). No asexual morph has been reported for Misturatosphaeria uniseriata (Mugambi and Huhndorf 2009). The sexual morph of Curreya austroafricana and C. proteae, however, share similar characteristics with those of Misturatosphaeria uniseriata Therefore, we introduced a new genus, Neocurreya for Curreya austroafricana, C. grandicipis and C. proteae in Floricolaceae based on morphology and phylogeny. Type species: Floricola striata Kohlm. & Volkm.-Kohlm., Bot. Mar. 43(4): 385 (2000); Fig. 28 Misturatosphaeria Mugambi & Huhndorf, in Schoch et al., Stud. Mycol. 64: 108 (2009) Facesoffungi number: FoF00831 Saprobic on woody branches. Sexual morph: Ascomata solitary or aggregated, superficial, coriaceous to carbonaceous, dark brown to black, pyriform or globose to subglobose, ostiolate, ostiolar area orange or lighter. Ostiole central, rounded, papillate. Peridium comprising several layers of cells, outer layers of dark-brown, compressed cells of textura angularis; becoming thin-walled and hyaline towards the inner region. Hamathecium comprising 1–2μm wide, numerous, septate, branching, anastomosing pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-


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Fig. 28 Floricola striata (holotype, redrawn from Kohlmeyer and Volkmann-Kohlmeyer 2000). a. Conidiomata wall and conidiogenesis cells b. Conidia. Scale bars: a, b=10μm

spored, bitunicate, fissitunicate, cylindrical-clavate, long pedicellate, apically rounded, with an occular chamber. Ascospores uni to bi-seriate, partially overlapping, brown to dark brown, fusiform, often slightly curved, 1–3-septate, one of the central cells is often wider than the others, slightly constricted at the central septum, rough-walled, with or without a mucilaginous sheath. Asexual morph: Undetermined. Type species: Misturatosphaeria aurantiinotata Mugambi & Huhndorf [as ‘aurantonotata’], Stud. Mycol. 64: 108 (2009) Facesoffungi number: FoF00832; Fig. 29 Saprobic on woody branches. Sexual morph: Ascomata 500–685 high×475–540μm diam. (x = 630×501μm, n=5), solitary or aggregated, superficial, coriaceous to carbonaceous, dark brown to black, pyriform or globose to subglobose, associated with a subiculum, ostiolate, ostiolar area orange. Ostiole central, rounded, papillate. Peridium 60–90 (x = 71 μm, n=15) μm wide, comprising several layers of cells; outer layers of dark-brown, compressed cells of textura angularis; becoming thin-walled and hyaline towards the inner region. Hamathecium comprising 1–2μm wide, numerous, septate, branching, anastomosing pseudoparaphyses, embedded in a gelatinous matrix. Asci 110–170×8.5–12μm (x = 130×10.2μm, n=10), 8-spored, bitunicate, fissitunicate, cylindrical-clavate, long pedicellate, apically rounded. Ascospores 18–22.5×5–6.5μm (x = 20.7×5.8μm, n=15) uni to bi-seriate, partially overlapping, fusiform, often slightly curved, at first hyaline, later becoming brown to dark brown, 3-septate at maturity, one of the central cells is often wider than the others, slightly constricted at the central septum, rough-walled, mucilaginous sheath present when the spores are young. Asexual morph: Undetermined. Material examined: KENYA, Rift Valley Province, Kajiando District, Ngong Hills Forest Reserve, on woody branch, 12 July 2006, G.K.Mugambi (GKM1238, isotype).

Notes: Misturatosphaeria was established by Mugambi and Huhndorf (2009), from Africa and North, Central and South America. Zhang et al. (2013) introduced M. mariae from Europe. In our phylogenetic analyses (Fig. 1) Misturatosphaeria species grouped in several subclades in Floricolaceae. The morphology and phylogeny of these species suggest new genera for these groups. Misturatosphaeria aurantiinotata Mugambi & Huhndorf, the type species of Misturatosphaeria and M. uniseptata are retained in Misturatosphaeria. The type strain of Melanomma radicans (ATCC 42522) clustered together with Misturatosphaeria uniseptata with 98 % ML and 0.96 B.P. (Fig. 1). Comparing illustration and description of M. radicans (NY!, holotype) in Khashnobish and Shearer (1996) with M. uniseptata in Mugambi and Huhndorf (2009), it seems that these two species are conspecific. Therefore, we introduce Misturatosphaeria radicans (Samuels & E. Müll.) Thambugala & K.D. Hyde, comb. nov. and synonymies Misturatosphaeria uniseptata and Melanomma radicans under Misturatosphaeria radicans. Misturatosphaeria radicans (Samuels & E. Müll.) Thambugala & K.D. Hyde, comb. nov. Basionym: Melanomma radicans Samuels & E. Müll., Sydowia 31(1–6): 147 (1979) [1978] ≡ Misturatosphaeria uniseptata Mugambi, A.N. Mill. & Huhndorf, in Mugambi & Huhndorf, Stud. Mycol. 64: 114 (2009) Index Fungorum number: IF551536 Asymmetrispora Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551259; Facesoffungi number: FoF00833 Etymology: In reference to its asymmetrical ascospores. Saprobic on dead branches. Sexual morph: Ascomata solitary, scattered, or in small groups, semi-immersed or

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Fig. 29 Misturatosphaeria aurantiinotata (isotype). a. Herbarium material b, c. Appearance of ascomata on host surface d–e. Section through ascomata f. Peridium g. Hamathecium h–j. Bitunicate asci k–n. Ascospores. Scale bars: d= 100μm, e=150μm, f=10μm, g, k–n=5μm, h–j=40μm

erumpent to nearly superficial, coriaceous to carbonaceous, dark brown to black, globose to subglobose or pyriform, with rounded to conical opening. Ostiole central, rounded, usually raised, papillate. Peridium thick at the apex, thinner at the base, comprising two cell types, outer layer composed of small heavily pigmented, thickwalled, compressed cells, inner layer thin, composed of smaller, hyaline cells of textura angularis. Hamathecium comprising 1–2 μm wide, numerous, septate, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical to cylindro-clavate, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores uni to bi-seriate, partially overlapping, pale to dark brown, fusiform to broadly clavate, with somewhat tapering lower end, 3-septate or muriform, asymmetrical, usually constricted at the central septum, smooth-walled, lacking a

mucilaginous sheath. Asexual morph: Undetermined (Mugambi and Huhndorf 2009; Zhang et al. 2013). Type species: Asymmetrispora tennesseensis (Mugambi, A.N. Mill. & Huhndorf) Thambugala & K.D. Hyde comb. nov. Basionym: Misturatosphaeria tennesseensis Mugambi, A.N. Mill. & Huhndorf, in Mugambi & Huhndorf, Stud. Mycol. 64: 114 (2009) Index Fungorum Number: IF551260 Asymmetrispora mariae (Ying Zhang, J. Fourn. & K.D. Hyde) Thambugala & K.D. Hyde comb. nov. Basionym: Misturatosphaeria mariae Ying Zhang, J. Fourn. & K.D. Hyde, Mycoscience 54(2): 2 (2012) Index Fungorum Number: IF551261 Notes: We introduce a new genus Asymmetrispora in Floricolaceae based on a unique combination of morphology and phylogeny with Misturatosphaeria tennesseensis as the


type species. In our phylogenetic analyses (Fig. 1), A. mariae grouped together with A. tennesseensis with 92 ML and 0.98 B.P. supports in subclade C2. Asymmetrispora mariae is therefore included in the genus, but mainly differs from A. tennesseensis, in having globose to subglobose ascomata without raised papillae, and dark brown, muriform ascospores, with (3–) 4 transverse septa (Zhang et al. 2013). It will be interesting to establish this genus with further collections in odrer to resolve the taxonomic relationship between A. mariae and A. tennesseensis, as there are significant morphological differences between them. Asymmetrispora can be distinguished from Misturatosphaeria in having semi-immersed to erumpent, globose to pyriform ascomata, without a subiculum, and pale to dark brown, fusiform to broadly clavate, muriform, smooth-walled, ascospores lacking a mucilaginous sheath (Mugambi and Huhndorf 2009; Zhang et al. 2013). Aurantiascoma Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551262; Facesoffungi number: FoF00834 Etymology: The generic name Aurantiascoma, refers to the orange pigment produced at the ascomata apex. Saprobic on dead branches. Sexual morph: Ascomata solitary or aggregated in small clusters, erumpent to superficial, pyriform to subglobose, dark brown, ostiolate, ostiolar area orange. Ostiole central, rounded, usually raised, papillate. Hamathecium comprising 1–2μm wide, numerous, branched, septate, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical-clavate, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores uni to bi-seriate, partially overlapping, hyaline, fusiform, 1-(–3)-septate, one of the central cells broader than others, constricted at the central septum, smooth-walled, surrounded by a thin mucilaginous sheath that extends at the apex of the spores. Asexual morph: Undetermined (Mugambi and Huhndorf 2009). Type species: Aurantiascoma minimum (Mugambi, A.N. Mill. & Huhndorf) Thambugala & K.D. Hyde, comb. nov. Basionym: Misturatosphaeria minima Mugambi, A.N. Mill. & Huhndorf, in Mugambi & Huhndorf, Stud. Mycol. 64: 114 (2009) Index Fungorum Number: IF551263 Notes: Misturatosphaeria minima shares a similar morphology with the type species of Misturatosphaeria, M. aurantiinotata in having superficial ascomata, with an orange ostiolar area, septate, cellular pseudoparaphyses and fusiform ascospores, but differs as M. aurantiinotata has pyriform ascomata forming on a brown subiculum and 3-septate, brown to dark brown, rough-walled ascospores. In our phylogenetic analyses (Fig. 1), M. minima formed a separate clade in Floricolaceae (clade C9). A new genus Aurantiascoma is therefore, introduced to accommodate M. minima. Misturatosphaeria kenyensis and M. minima are quite similar

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in their morphologies, but the latter differs in its ascomata, which are solitary or in small groups of usually less than five individuals and slightly narrower ascospores (Mugambi and Huhndorf 2009). Magnibotryascoma Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551266; Facesoffungi number: FoF00835 Etymology: In reference to occurrence of large clusters of ascomata on the host surface. Saprobic on dead branches. Sexual morph: Ascomata solitary, or aggregated in large clusters, erumpent, pyriform to subglobose, dark brown, ostiolate, ostiolar area often lighter. Ostiole central, rounded, slightly raised, papillate, with a porelike opening. Hamathecium comprising 1–2μm wide, numerous, branched, septate, hyaline, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical to cylindrical-clavate, pedicellate, rounded at the apex, with an ocular chamber. Ascospores uniseriate, partially overlapping, brown to dark brown, fusiform to elliptical, usually 1–3-septate, but commonly 3-septate, constricted at the septa, guttulate, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined (Mugambi and Huhndorf 2009). Type species: Magnibotryascoma uniseriatum (Mugambi, A.N. Mill. & Huhndorf) Thambugala & K.D. Hyde, comb. nov. Basionym: Misturatosphaeria uniseriata Mugambi, A.N. Mill. & Huhndorf, in Mugambi & Huhndorf, Stud. Mycol. 64: 116 (2009) Index Fungorum Number: IF551267 Notes: Magnabotrioscoma is introduced to accommodate Misturatosphaeria uniseriata which has unique morphological characters in Floricolaceae. Magnabotrioscoma differs from Neocurreya which is the phylogenetically most closely related genus (Fig. 1) in having erumpent to superficial ascomata, with short, rounded prominent openings that appear plugged by centrum tissue and uniseriate, partially overlapping, fusiform to elliptical, 3-septate, guttulate ascospores. Misturatosphaeria (Asymmetrispora) tennesseensis has some similar morphological characteristics to M. uniseriata, but differs in the shape and loose aggregation of the ascomata and paler brown ascospores, which are strongly constricted at the central septum. Neocurreya Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551268; Facesoffungi number: FoF00836 Etymology: In reference to its similarity with Curreya. Saprobic on twigs and branches or leaves. Sexual morph: Ascomata solitary or aggregated, immersed, becoming erumpent to superficial, coriaceous, subglobose or pyriform to obclavate, dark brown, with broad, rounded opening, with a long neck, ostiolate. Ostiole central, central, periphysate, with a pore-like opening. Peridium comprising 2-layers, outer layer

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composed of brown, thick-walled cells, inner layer consisting of a few layers of hyaline, thin-walled, compressed cells of textura angularis. Hamathecium comprising 1–2μm wide, branched, septate, filamentous, anastomosing pseudoparaphyses, embedded in a gelatinous matrix. Asci bitunicate, fissitunicate, cylindrical, pedicellate, apically rounded, with an ocular chamber, slightly elongated or inflated at the apex. Ascospores uni to bi-seriate, brown to dark brown, elliptical, broadly fusiform or ovoid, with the one end pointed and the other obtuse, muriform, 3–4(–5)-transversely septate, with one vertical septum in median cells, end cells without septa or with a Y-form pair of septa, one longitudinal or oblique septum, smooth-walled, lacking a mucilaginous sheath (Mugambi and Huhndorf 2009; Marincowitz et al. 2008). Asexual morph: Conidiomata pycnidial, subepidermal, substromatal, dark brown to black, globose or subglobose, ostiolate. Conidiomata wall several-layered, hyaline to brown cells of textura angularis. Conidiophores hyaline, 0–2-septate and branched at the base. Conidiogenous cells discrete, smooth, indeterminate, cylindrical, hyaline to pale brown, branched, conidiogenesis holoblastic, proliferating 1–3 times precurrently, arising from the inner cells of the pycnidial wall. Conidia ellipsoidal or ellipsoidal to oblong, aseptate, pale to dark brown, smooth-walled (Asexual morph follows Coniothyrium grandicipis in Crous et al. (2004). Type species: Neocurreya austroafricana (Marinc., M.J. Wingf. & Crous) Thambugala & K.D. Hyde, comb. nov. Basionym: Curreya austroafricana Marinc., M.J. Wingf. & Crous, in Marincowitz et al., CBS Diversity Ser. (Utrecht) 7: 37 (2008) Index Fungorum number: IF551269 Notes: Curreya austroafricana and C. proteae were introduced by Marincowitz et al. (2008), based on their morphological similarities with Curreya, while C. grandicipis (Coniothyrium grandicipis Joanne E. Taylor & Crous) was introduced by Crous et al. (2011) as coniothyrium-like asexual morphs reported in species of Curreya. However, this classification is doubtful, as in our phylogenetic analyses they clustered in our new family Floricolaceae together with Misturatosphaeria claviformis. These species formed a separate clade (C7) in Floricolaceae. This group of species show unique morphological characters as compared to other genera in Floricolaceae, such as immersed to superficial, pyriform to obclavate ascomata, with long necks and periphysate ostioles, filamentous pseudoparaphyses, and brown to dark brown, broadly fusiform or ovoid to elliptical, muriform ascospores without a mucilaginous sheath. Considering the morphological and phylogenetic affinities we accommodate C. austroafricana, C. proteae and Misturatosphaeria claviformis in a new genus Neocurreya in Floricolaceae. Neocurreya austroafricana and N. proteae are morphologically similar, except for the larger ascospores in C. proteae.


Neocurreya claviformis (Mugambi & Huhndorf) Thambugala & K.D. Hyde, comb. nov. Basionym: Misturatosphaeria claviformis Mugambi & Huhndorf, Stud. Mycol. 64: 113 (2009) Index Fungorum Number: IF551272 Neocurreya grandicipis (Joanne E. Taylor & Crous) Thambugala & K.D. Hyde, comb. nov. Basionym: Coniothyrium grandicipis Joanne E. Taylor & Crous, in Crous et al., CBS Diversity Ser. (Utrecht) 2: 60 (2004) ≡ Curreya grandicipis (Joanne E. Taylor & Crous) Joanne E. Taylor & Crous, in Crous et al. Persoonia, Mol. Phyl. Evol. Fungi 27: 32 (2011) Index Fungorum number: IF551271 Neocurreya proteae (Marinc., M.J. Wingf. & Crous) Thambugala & K.D. Hyde, comb. nov. Basionym: Curreya proteae Marinc., M.J. Wingf. & Crous, in Marincowitz et al., CBS Diversity Ser. (Utrecht) 7: 39 (2008) Index Fungorum number: IF551270 Pseudoaurantiascoma Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551273; Facesoffungi number: FoF00837 Etymology: In reference to its similarity w ith Aurantiascoma. Saprobic on dead branches. Sexual morph: Ascomata solitary or aggregated into large clusters or in small groups, erumpent to superficial, pyriform to globose, dark brown, ostiolar area often lightly coloured. Ostiole central, rounded, usually raised, papillate, with a pore-like opening. Hamathecium comprising 1–2μm wide, numerous, branched, septate, cellular, pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical-clavate, pedicellate, rounded at the apex, with an ocular chamber. Ascospores bi-seriate, partially overlapping, hyaline, fusiform, usually 1–3-septate, but commonly 1-septate, occasionally one of the central cells broader than others, constricted at the central septum, smooth-walled, surrounded by a small mucilaginous sheath that extends at the apex of ascospores. Asexual morph: Undetermined (Mugambi and Huhndorf 2009). Type species: Pseudoaurantiascoma kenyense (Mugambi & Huhndorf) Thambugala & K.D. Hyde, comb. nov. Basionym: Misturatosphaeria kenyensis Mugambi & Huhndorf, Stud. Mycol. 64: 113 (2009) Index Fungorum Number: IF551274 Notes: Pseudoaurantiascoma is introduced to accommodate Misturatosphaeria kenyensis based on both morphological and molecular characters. Pseudoaurantiascoma kenyense differs from the type species of Misturatosphaeria in having 1–3-septate ascospores, surrounded by a small mucilaginous sheath, that extends at the ends of ascospores. In our phylogenetic analyses the ex-type strain of M. kenyensis formed a


separate subclade (C4). Hence we introduce it as a new genus in Floricolaceae. Pseudomisturatosphaeria Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551275; Facesoffungi number: FoF00838 Etymology: In reference to its similarity to Misturatosphaeria. Saprobic on decorticated woody branches. Sexual morph: Ascomata usually single or rarely clustered in small groups, erumpent, dark brown, pyriform to globose, ostiolate. Ostiole rounded, papillate, usually raised. Hamathecium comprising 1–2μm wide, numerous, septate, branching, anastomosing, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical-clavate, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores partially bi-seriate to sometimes overlapping uniseriate, hyaline when immature, becoming brown to dark brown when mature, oblong to elliptical, muriform, usually constricted at the central transverse septum, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined (Mugambi and Huhndorf 2009). Type species: Pseudomisturatosphaeria cruciformis (Mugambi & Huhndorf) Thambugala & K.D. Hyde comb. nov. Basionym: Misturatosphaeria cruciformis Mugambi & Huhndorf, Stud. Mycol. 64: 113 (2009) Index Fungorum Number: IF551276 Notes: Misturatosphaeria cruciformis was introduced by Mugambi and Huhndorf (2009) and it shares a similar ascospore morphology with Misturatosphaeria (Neocurreya) claviformis Mugambi & Huhndorf, but the latter differs in having much larger ascomata that are predominantly pyriform and ascospores that are often constricted at the central septum (Mugambi and Huhndorf 2009). The type species of Misturatosphaeria, M. aurantiinotata however, is distinct from M. cruciformis in having a sparse, brown subiculum, an orange ostiolar area and fusiform, often slightly curved, hyaline to brown or dark brown, 3-septate, rough-walled ascospores, surrounded by a mucilaginous sheath. In the present phylogenetic analysis M. cruciformis clustered as a sister clade (C3) to Asymmetrispora (Fig. 1). Misturatosphaeria cruciformis differs from Asymmetrispora species in having ascomata usually occurring singly and oblong to elliptical, symmetrical, dictyosporous ascospores with 5-7-transverse septa. Therefore, considering the substantial differences in morphology and distinct differences in phylogeny we introduced a new genus Pseudomisturatosphaeria for M. cruciformis in the family Floricolaceae. Ramusculicola Thambugala & K.D. Hyde, gen. nov. Index Fungorum number: IF551264; Facesoffungi number: FoF00839 Etymology: referring to its habitat on twigs.

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Saprobic on dead twigs in terrestrial habitats. Sexual morph: Ascomata solitary or scattered, immersed, papilla usually erumpent, black, subglobose to globose, coriaceous, uniloculate, ostiolate. Ostiole central, rounded, periphysate, with a pore-like opening. Peridium 1-layered, composed of small, dark brown to lightly pigmented cells of textura angularis, fusing at the outside with the host tissues. Hamathecium comprising 1–2μm wide, numerous, filamentous, aseptate, sparsely anastomosing, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 8spored, bitunicate, fissitunicate, cylindrical, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores overlapping bi-seriate, hyaline, fusiform to cylindrical, usually 1-septate, constricted at the septum, surrounded by a thin mucilaginous sheath, filled with small guttules when immature. Asexual morph: Undetermined. Type species: Ramusculicola thailandica Thambugala & K.D. Hyde sp. nov. Index Fungorum Number: IF551265; Facesoffungi number: FoF 01092; Fig. 30 Etymology: Named after the country, where it was collected, Thailand. Saprobic on dead twigs in terrestrial habitats. Sexual morph: Ascomata 190–260 μm high, 190–280 μm (x = 222 × 233 μm, n = 10) diam., solitary or scattered, immersed, papilla usually erumpent, black, subglobose to globose, coriaceous, uniloculate, ostiolate. Ostiole central, rounded, compressed, periphysate, with a pore-like opening. Peridium 19–44μm (x = 29μm, n=10) wide, 1-layered, composed of small, dark brown to lightly pigmented cells of textura angularis, fusing at the outside with the host tissues. Hamathecium comprising 1–2μm wide, numerous, filamentous, aseptate, sparsely anastomosing, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 55–75×8–10μm (x = 62.5×9.3μm, n=15), 8-spored, bitunicate, fissitunicate, cylindrical, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores 18–27×3.6–5μm (x = 22.5×4.3μm, n=20), overlapping bi-seriate, hyaline, fusiform to cylindrical, usually 1-septate, constricted at the septum, surrounded by a thin mucilaginous sheath, filled with small guttules when immature. Asexual morph: Undetermined. Culture characteristics: Ascospores germinating on PDA within 18 h and producing germ tubes from one or both ends. Colonies on PDA attaining 20 mm diam. after 14 days at 25– 30 °C, flat, circular, dull red to pink, reverse dull red, with entire to slightly undulate margin. Material examined: THAILAND, Chiang Rai Province, Mae Fah Luang University Garden, on fallen twig of deciduous tree, 12 January 2013, Kasun M. Thambugala TL 031 (MFLU 14-0587, holotype); ibid., (HHUF 30459, isotype); ex-type living culture, MFLUCC 13-0284, ICMP 20756; ibid., 10 June 2009, Saranyaphat Boonmee DT-03

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Fig. 30 Ramusculicola thailandica (holotype). a, b. Appearance of ascomata on the host surface c. Section through ascoma d. Ostiole with periphyses e. Section through peridium f Pseudoparaphyses g–i. Immature and mature bitunicate asci j–l. Released ascospores with thin sheath m. Germinating ascospores Scale bars: c=100μm, d=50μm, e, f=25μm, g-i=20μm, j–m=10μm

(MFLU10-0059, paratype), living culture, MFLUCC100126, BCC 38862, IFRD 2182. Notes: Ramusculicola is introduced to accommodate our new collection from Thailand. Ramusculicola differs from other genera in Floricolaceae in having semi-immersed to partially erumpent, coriaceous ascomata, a comparatively thin-walled peridium and short pedicellate asci. Ramusculicola separates distinctly from other genera in the phylogenetic tree (Fig. 1) and can be considered as a new genus. Asci and ascospores characters of Ramusculicola thailandica share similar ascospore morphology with some other genera in Floricolaceae such as Aurantiascoma and Pseudoaurantiascoma. Amorosiaceae Thambugala, K. D. Hyde, fam. nov.

Index Fungorum number: IF551277; Facesoffungi number: FoF01084 Fungicolous or saprobic on other fungi or dead wood. Sexual morph: Ascomata solitary or gregarious, immersed to semiimmersed, becoming erumpent, coriaceous, dark brown to black, globose to subglobose or conical, uniloculate, ostiolate. Ostiole crest-like, central, rounded, cylindrical, papillate. Peridium comprising several layers of dark brown to lightly pigmented cells of textura angularis, fusing with the stromata. Hamathecium comprising numerous, wide, cellular, septate, unbranched pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical to cylindricclavate, pedicellate, rounded at the apex, with an ocular chamber. Ascospores uni to tri-seriate, partially overlapping, hyaline,


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some light brown when mature, fusiform, to cylindrical, or ellipsoidal-fusiform, 1(–3)-septate, constricted at the central septum, filled with small guttules when immature, surrounded by a mucilaginous sheath. Asexual morph: hyphomycetous. Chlamydospores occasionally, formed in short chains, arising from the mycelium, individual chlamydospores subhyaline, broadly ellipsoid to subglobose. Conidiophores micronematous to semimacronematous, arising singly and not combined in sporodochia or synemmata, pale brown, unbranched, similar to the mycelium. Conidiogenous cells integrated, terminal or intercalary, where terminal, monoblastic, determinate, shortcylindrical to elongate-cylindrical, subhyaline to pale brown or brown, smooth-walled. Conidia solitary, dry, lateral, elongate-clavate, pale brown to brown, 1–3-septate, 3–4-septate when mature, constricted at the septa, smooth-walled, lacking any mucilaginous sheath or appendages. Type: Amorosia Mantle & D. Hawksw., in Mantle et al., Mycol. Res. 110(12): 1373 (2006) Type species: Amorosia littoralis Mantle & D. Hawksw., in Mantle et al., Mycol. Res. 110(12): 1373 (2006); Fig. 31 Notes: The hyphomycete genus Amorosia was introduced from intertidal sediment in the Bahamas Islands by Mantle et al. (2006) to accommodate A. littoralis Mantle & D. Hawksw. Mantle et al. (2006) placed this genus in Sporormiaceae based on molecular phylogenetic analysis. Amorosia is characterized by micronematous to semimacronematous conidiophores, integrated, terminal or intercalary conidiogenous cells, elongate-clavate and 3–4-septate conidia with a distinct central pore in each septum. In our multi-gene phylogenetic analysis the ex-type strain of

A. littoralis clustered in subclade E2 as a sister clade to the Angustimassarina clade (E1). This clade (E) forms a wellsupported clade in between clade D and Sporormiaceae. Therefore, we believe that Amorosia should not be placed in Sporormiaceae and a new family Amorosiaceae is proposed for Amorosia and Angustimassarina. Amorosiaceae mainly differs from Lophiostomataceae, Floricolaceae and Sporormiaceae in having hyphomycete asexual morphs and appears to grow within other ascomata of other ascomycetes and may be mycoparasitic. Angustimassarina Thambugala, Kaz. Tanaka & K.D. Hyde, gen. nov. Index Fungorum Number: IF551278; Facesoffungi number: FoF01085 Etymology: angustus (arrow)+Massarina, referring to narrowly fusiform ascospores. Fungicolous or saprobic on other fungi or dead wood in terrestrial habitats. Sexual morph: Ascomata solitary or gregarious, immersed to semi-immersed, becoming erumpent, coriaceous, dark brown to black, globose to subglobose, uniloculate, ostiolate. Ostiole central, cylindrical, papillate, and composed of pseudoparenchymatous cells, broad, well-developed, with a pore-like opening or through the cracks of host surface. Peridium composed of several layers of dark brown to lightly pigmented cells of textura angularis, fusing at the outside with the host ascomata. Hamathecium comprising 1–2.5μm wide, septate, unbranched, cellular, pseudoparaphyses, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindrical to cylindric-clavate, short

Fig. 31 Amorosia littoralis (holotype, redrawn from Mantle et al. 2006). a. Chlamydospores b–d. Terminally produced conidiogenous cells and conidia e, f. Laterally produced conidia and conidiogenous cells g–j.

Mature released 3–4-septate conidia k. Conidium in which the apical cell is germinating. Scale bar=10μm

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pedicellate, rounded at the apex, with an ocular chamber. Ascospores uni to tri-seriate, partially overlapping, hyaline, becoming ocher brown at maturity, fusiform to cylindrical or ellipsoidal-fusiform, mostly straight, widest at the centre and tapering toward the ends, 1(–3)-septate, constricted at the central septum, smooth-walled, verruculose at maturity, filled with a different sized guttule per cell and surrounded by a mucilaginous sheath. Asexual morph: hyphomyc e t o u s . C o n i d i o p h o re s m i c r o n e m a t o u s t o s e m i macronematous, arising singly and not in sporodochia or synemmata, pale brown, unbranched, similar to the mycelial hypha. Conidiogenous cells integrated, terminal, holoblastic, short-cylindrical to elongate-cylindrical, brown to pale brown, smooth-walled. Conidia solitary, dry, elongateFig. 32 Angustimassarina populi (holotype). a, b. Ascomata immersed on host surface c. Section through ascomata (immersed in host fungi) d. Peridium e. Pseudoparaphyses f–h. Mature bitunicate asci i–k. Ascospores l. Germinating ascospore. Scale bars: c=100μm, d–h=20μm, i– l=10μm


clavate, pale to dark brown, 1–3-septate, evenly pigmented, constricted at the septa, smooth-walled. Notes: We introduce Angustimassarina to accommodate three new ascomycetous species from Italy and Germany. The Angustimassarina clade (E1) clustered as a sister clade to the Amorosia (E2) clade in Amorosiaceae (E). Species of Angustimassarina are considered as fungicolous. They may be parasitic on other fungi and appear to grow within other ascomata of other ascomycetes. The Angustimassarina clade formed a distinctly separate sister clade to the Amorosia clade with high ML and BYPP support. Type species: Angustimassarina populi Thambugala & K.D. Hyde, sp. nov.


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Fig. 33 Angustimassarina populi (Asexual morph from the ex-type living culture). m, n. Culture on PDA o–r. Conidiogenesis and developing conidia. Scale bars: o–s=10μm

Index Fungorum Number: IF551279; Facesoffungi number: FoF 01086; Figs. 32 and 33 Etymology: Referring to the host genus Populus. Holotype: MFLU 14-0588 Fungicolous or saprobic on other fungi or Populus sp. in terrestrial habitats. Sexual morph: Ascomata 125– 175 μm high, 100–120 μm (x = 154 × 137.8 μm, n = 10) diam., solitary or gregarious, immersed, becoming erumpent, coriaceous, black, globose to subglobose, uniloculate, ostiolate. Ostiole crest-like, rounded, papillate, immersed in other ascomata, with a pore-like opening. Peridium 14–32μm (x = 23μm, n=15) wide, composed dark brown to hyaline cells of textura angularis. Hamathecium comprising 1–2 μm wide, septate, unbranched, cellular pseudoparaphyses, embedded in a gelatinous matrix. Asci 80–95×9.5–13μm (x = 87×11.4μm, n=15), 8-spored, bitunicate, fissitunicate, cylindrical, pedicellate, rounded at the apex, with an ocular chamber. Ascospores 19–22×3.2–5.5μm (x = 20.5×4.3μm, n=20), uni to bi-seriate, partially overlapping, hyaline, fusiform, 1(–3)-septate, constricted at the central septum, filled with a different sized guttule per cell and surrounded by a mucilaginous sheath. Asexual morph: hyphomycetous. Colonies on PDA moderately growing, circular with a wavy margin, cottony on the surface, grayish brown, with reverse dark brown. Conidiophores 2.5– 4μm wide, micronematous to semi-macronematous, arising singly and not in sporodochia or synemmata, pale brown, unbranched, similar to the mycelial hypha. Conidiogenous cells 4.2–9.8 × 2.3–3.4 μm (x = 6.9 × 2.9μm, n= 10), integrated, terminal, holoblastic, shortcylindrical to elongate-cylindrical, brown to pale brown,

smooth-walled. Conidia 6.6–10×5.6–7.5μm (x = 8.3× 6.5 μm, n = 10) solitary, dry, elongate-clavate, pale to dark brown, 1–3-septate, evenly pigmented, constricted at the septa, smooth-walled. Culture characteristics: Ascospores germinating on PDA within 18 h and producing germ tubes from both ends. Colonies on PDA attaining 40 mm diam. after 5 weeks at 25 °C, flat, grayish brown, with reverse dark brown, with crenate edge. After 2 weeks the culture produces lightly pigmented chemical exudate. Mycelium well-developed, superficial, composed of pale brown, thin-walled hyphae, frequently septate, smooth-walled. Chlamydospores sometimes formed individually, pale brown, broadly globose to subglobose. Material examined: ITALY, Province of Forlì-Cesena [FC], Strada San Zeno, Galeata, on dead branches of Populus sp., 15 November 2012, Erio Camporesi IT 896 (MFLU 14-0588, holotype); ibid., (HHUF 30460, isotype); ex-type living culture, MFLUCC 13-0034, ICMP 20742. Angustimassarina acerina Jayasiri, Thambugala, R.K. Schumach. & K.D. Hyde, sp. nov. Index Fungorum Number: IF551280; Facesoffungi number: FoF01087; Fig. 34 Etymology: Referring to the host genus Acer. Holotype: MFLU 14-0611 Fungicolous or saprobic on other fungi or dead wood. Sexual morph: Ascomata 200–350 × 164–183 μm (x = 280× 170μm, n=5), scattered to gregarious, immersed, coriaceous, dark brown to black, globose to subglobose, without a crest, ostiolate. Ostiole central, rounded, papillate, opening through the cracks of host surface. Peridium 15–26 μm (x = 20 μm, n = 10) thick at the

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Fig. 34 Angustimassarina acerina (holotype). a, b. Ascomata immersed in the host tissue c. Section through ascoma d. Ostiole e. Peridium f. Pseudoparaphyses g–i. Mature and immature asci j–m. Ascospores (m. Ascospore mounted in Indian ink) n. Germinated ascospore. Scale bars: a, b=500μm, c=100μm, d= 30μm, e, g–i=20μm, f, j–n= 10μm

sides, broad at the apex and thinner at the base, comprising brown to dark brown cells of textura angularis, fusing at the outside with the host tissues. Hamathecium comprising 1.5–2.5 wide, septate, long, cellular pseudoparaphyses, embedded in a gelatinous matrix.

Asci 92–105 × 7.5–8.6 μm (x = 97 × 8 μm, n = 10), 8spored, bitunicate, fissitunicate, cylindrical to clavate, pedicellate, rounded at the apex, with a minute ocular chamber. Ascospores 21–23 × 4.1–4.6 μm (x = 21 × 4.2μm, n=15), bi-seriate, partially overlapping, hyaline,


becoming ocher brown at maturity, 1(–3)-septate, deeply constricted at the central septum, widest at the centre and tapering toward the ends, straight, smooth-walled, verruculose at maturity, guttulate, surrounded by a thin mucilaginous sheath. Asexual morph: Undetermined. Material examined: GERMANY, on twigs of Acer platanoides L. (Sapindaceae), 23 February 2014, René K. Schumacher G005 (MFLU 14-0611, holotype); ibid., (HHUF 30456, isotype); ex-type living cultures, MFLUCC 14-0505, ICMP 20711. Culture characteristics: Ascospores germinating on PDA within 24 h and germinating from one end. Colonies on PDA attaining 35 mm diam. after 24 days at 25 °C, flat to effuse, Fig. 35 Angustimassarina quercicola (holotype). a, b. Ascomata immersed on host surface c. Section through ascoma d. Peridium e. Pseudoparaphyses f, g. Mature bitunicate asci h–k. Ascospores (k. Ascospore stained with Indian ink). Scale bars: c= 100μm, d=20μm, e, h–k=10μm, f, g=40μm

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circular, grey, reverse greyish white, with entire to slightly undulate edge. After 7–14 days culture producing exudates (Fig. 19). Notes: Angustimassarina acerina and A. quercicola are morphologically similar species, but they occur on different hosts and are phylogenetically different (Fig. 1). Angustimassarina quercicola differs from A. acerina in having a peridium of somewhat flattened cells of textura angularis and wider ascospores surrounded by a wide mucilaginous sheath. Angustimassarina quercicola Thambugala, Jayasiri, R.K. Schumach. & K.D. Hyde, sp. nov. Index Fungorum Number: IF551281; Facesoffungi number: FoF01088; Fig. 35

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Etymology: Referring to the host genus Quercus. Holotype: MFLU 14-0612 Saprobic on dead wood, often forming fruiting bodies in old ascomata of other fungi, possible fungicolous. Sexual morph: Ascomata 200–250 high×150–265μm (x = 224× 221μm, n=5) diam., scattered to gregarious, immersed, coriaceous, dark brown to black, globose to subglobose, ostiolate. Ostiole central, rounded, papillate, with a pore-like opening. Peridium 14–27μm (x = 19.7μm, n=15) thick at the sides, broad at the apex and thinner at the base, comprising of brown to dark brown to lightly pigmented cells of textura angularis, fusing with the host tissue. Hamathecium comprising 1–2μm wide, septate, cellular pseudoparaphyses, embedded in a gelatinous matrix, anastomosing above the asci. Asci 60–94× 8.8–13μm (x = 77×11.2μm, n=15), 8-spored, bitunicate, fissitunicate, cylindrical to clavate, with a short pedicel, rounded at the apex, with an ocular chamber. Ascospores 17–21×4–6μm (x = 19.5×4.6μm, n=20), bi-seriate, partially overlapping, hyaline, becoming ocher brown at maturity, 1(– 3)-septate, deeply constricted at the central septum, widest at the centre and tapering toward the ends, straight, guttulate, smooth-walled, finely verruculose at maturity, surrounded by a wide mucilaginous sheath. Asexual morph: Undetermined. Material examined: GERMANY, on twigs of Quercus robur L. (Fagaceae), 28 December 2013, Rene K. Schumacher G013 (MFLU 14-0612, holotype); ex-type living culture, MFLUCC 14-0506, ICMP 21070. Culture characteristics: Ascospores germinating on PDA within 24 h and producing germ tubes from one end. Colonies on PDA attaining 36 mm diam. after 24 days at 25 °C, flat to effuse, circular, greyish black, reverse white to greyish black,


with entire edge. After 1–2 week culture produce exudates (Fig. 36). Massarina corticola (Fuckel) L. Holm, Symb. bot. upsal. 14(no. 3): 147 (1957) ≡ Trematosphaeria corticola Fuckel, Jb. nassau. Ver. Naturk. 23–24: 162 (1870) [1869–70] Saprobic on palms and various other hosts. Sexual morph: Ascomata scattered to gregarious, immersed to semi-immersed, becoming erumpent, coriaceous, black, subglobose, base flattened, surrounded by darkened pseudostromatic tissue, ostiolate. Ostiole central, cylindrical, broadly papillate. Peridium comprising several layers of brown-walled, compressed cells, fusing with the stromata at the outside. Hamathecium comprising numerous, filamentous, pseudoparaphyses, anastomosing above the asci, embedded in a gelatinous matrix. Asci 8-spored, bitunicate, fissitunicate, cylindric-clavate, or clavate, short prdicellate, apically rounded with an ocular chamber. Ascospores 2–3 seriate, hyaline, ellipsoidal-fusiform, 1-septate, constricted at the septum, some light brown when mature, smooth-walled, with a thin mucilaginous sheath (Hyde and Aptroot 1997). Asexual morph: Undetermined. Notes: Trematosphaeria corticola was introduced by Fuckel (1870) and then Holm (1957) transferred this species to Massarina based on its morphology. Liew et al. (2002) transferred this species to Lophiostoma. This species is thought to be widely distributed and occurs on various hosts including palms (Hyde and Aptroot 1997). In our phylogenetic analysis a putative strain of M. corticola (CBS 154.93) grouped in the Angustimassarina clade (E) and can be another species

Fig. 36 Culture characteristics of a, b. = Angustimassarina acerina (MFLUCC 14-0505). c, d. = Angustimassarina quercicola (MFLUCC 14-0506)


of Angustimassarina. However, recollection and epitypification are needed to confirm the placement. Exosporium stylobatum Curzi & Barbaini, Atti Ist. bot. R. Univ. Pavia, 3 Sér. 3(3): 199 (1927) Notes: Exosporium is a hyphomycetous genus characterised by stroma that gives rise to fasciculate conidiophores with sympodial proliferation, and darkened scars, each with a visible central pore (Crous et al. 2011). In our phylogenetic analysis a putative strain of Exosporium stylobatum (CBS 160.30) clustered in the Angustimassarina clade along with M. corticola (CBS 154.93) and Angustimassarina populi (MFLU 140588). In Crous et al. (2011) Exosporium livistonae Crous & Summerell grouped in Capnodiales. Onehundred and twenty five epithets are listed as Exosporium in Index Fungorum (2015) and molecular sequence data are not available for the type species, Exosporium tiliae Link.

Dothideomycetes, genera incertae sedis Notes: The family Thyridariaceae Q. Tian & K.D. Hyde was introduced by Hyde et al. (2013) to accommodate the genus Thyridaria Sacc. based on its unique morphology and clustering in a unique family in the Dothideomycete phylogeny. The placement of Thyridaria has been assigned to various families such as Melanommataceae, Platystomaceae, Pleosporaceae and Sphaeriaceae by various authors (Barr 1979a, b; Luttrell 1973; Munk 1957; Müller and Arx 1973; Wehmeyer 1975). Thyridaria is characterized by perithecial, coriaceous, ascomata surrounded by a subiculum, cylindrical to subclavate asci and ellipsoid to fusiform, dark brown to blackish-brown, 2–3-septate ascospores (Hyde et al. 2013). In our phylogenetic analysis, a putative strain of Thyridaria macrostomoides (De Not.) M.E. Barr clustered in clade D, together with Decaisnella formosa Abdel-Wahab & E.B.G. Jones and our two new fungal strains (MFLUCC 11-0376 and MFLUCC 11-0377) which are described here in a new genus Lignosphaeria. Zhang et al. (2012) listed Decaisnella in Pleosporales genera, incertae sedis and currently 14 epithets are listed for the genus in Index Fungorum (2015). A putative strain of T. rubronotata (Berk. & Broome) Sacc. was included by Schoch et al. (2009) and Hyde et al. (2013) which clustered in a separate lineage in the order Pleosporales. Sequences from the type strains of Decaisnella and Thyridaria are needed to resolve their phylogenetic placement in Pleosporales in order to confirm whether they represent distinct families or not. Lignosphaeria Boonmee, Thambugala & K.D. Hyde, gen. nov. Index Fungorum Number: IF551282; Facesoffungi number: FoF01089 Etymology: Referring to its habitat on dead wood.

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Saprobic on dead wood. Sexual morph: Ascomata scattered to gregarious, immersed to erumpent, coriaceous to carbonaceous, black, subglobose, ostiolate. Ostiole slit-like, central, papillate, periphysate. Peridium thick, broad at the apex, comprising several layers of dark brown to black or hyaline cells of textura angularis, fusing at the outside with the host tissues. Hamathecium comprising 2–3μm wide, septate, filiform, unbranched, pseudoparaphyses. Asci 8-spored, bitunicate, fissitunicate, cylindrical to clavate, pedicellate, apically rounded with an ocular chamber. Ascospores uni to triseriate, hyaline, cylindrical to fusiform, narrowly rounded or acute at the ends, wide at the second cell from the apex, 3septate, constricted at the median septum, guttulate, smoothwalled, lacking a mucilaginous sheath. Asexual morph: coelomycetous. Conidiomata grouped to scattered, partly immersed, subglobose to globose, with a papillate ostiole, median brown. Conidiomatal wall thin, 2-layered, outer layer composed of dark brown to median brown cells of textura epidermoidea to intricate, with inner layer subhyaline. Conidiophores monochasial, cylindrical, straight or curved, septate, branched, hyaline, smooth, developed from cells of inner layer of conidiomatal wall. Conidiogenous cells holoblastic to phialidic, cylindrical, hyaline, with a conspicuous collarette at the tip. Conidia one-celled, oval-ellipsoid, with obtuse apex, widest in the middle, narrow and truncate at the base, aseptate, hyaline, guttulate, smooth-walled. Notes: We describe Lignosphaeria as a new genus based on morphology and multi-gene phylogeny to accommodate Lignosphaeria thailandica and L. fusispora. In our phylogenetic tree these two species grouped together in the clade D and form a well-supported sister clade (D3) (100 ML/1.00 BYPP) to T. macrostomum (D2). Lignosphaeria can be distinguished from Thyridaria (Hyde et al. 2013) in having immersed to erumpent ascomata, pseudoparaphyses and cylindrical to fusiform ascospores with narrowly rounded or acute ends. We include Lignosphaeria in Dothideomycetes, genera incertae sedis as there is no existing family to accommodate this new genus. Type species: Lignosphaeria thailandica Boonmee, Thambugala & K.D. Hyde, sp. nov. Index Fungorum Number: IF551283; Facesoffungi number: FoF01090; Figs. 37 and 38 Etymology: Named after the country, where it was collected, Thailand. Holotype: MFLU11-0135 Saprobic on dead wood. Sexual morph: Ascomata 390– 468μm high×299–315μm (x = 418.5×308μm, n=3) diam., scattered to gregarious, semi-immersed to erumpent, carbonaceous, black, subglobose, ostiolate. Ostiole slit-like, central, papillate, periphysate. Peridium 40–45μm (x = 58.5×59μm, n=6) wide, broad at the apex, comprising several layers of dark brown to black cells of textura angularis, fusing at the outside with the host tissues. Hamathecium comprising 2–3μm wide, septate, unbranched, cellular pseudoparaphyses. Asci 63–68× 7–8.5 μm (x = 65 × 8 μm, n = 10), 8-spored, bitunicate,

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Fig. 37 Lignosphaeria thailandica (holotype). a. Ascomata immersed on host surface b, c. Section of ascoma and peridium d. Pseudoparaphyses e, f. Asci g–j. Ascospores. Scale bars: a= 500μm, b=100μm, c=40μm, d= 5μm, e, f=20μm, g–j=10μm

cylindrical to clavate, long pedicellate, apically rounded, with a minute ocular chamber. Ascospores 15–19×5–6μm (x = 17× 5μm, n=10), uni to bi-seriate, hyaline, cylindrical to fusiform, with narrowly rounded ends, wide at the second cell from the apex, 3-septate, constricted at the septa, cell above central septum widest, small guttulate, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Conidiomata 162–335μm diam. × 143–300μm high, grouped to scattered, partly immersed, subglobose to globose, with a papillate ostiole, median brown. Conidiomatal wall thin, 13–15μm diam., 2-layered, outer layer composed of dark brown to median brown cells of textura epidermoidea to intricate, with inner layer subhyaline. Conidiophores 12–28×1.5–2μm monochasial, cylindrical, straight or curved, septate, branched, hyaline, smooth, developed from cells of inner layer of conidiomatal wall. Conidiogenous cells 2.5–3μm wide, holoblastic to phialidic, cylindrical, hyaline, with a conspicuous collarette at the apex. Conidia 3–4×2–3μm (x = 4×2μm, n=20), one-celled, ovalellipsoid, with obtuse apex, widest in the centre, narrow and truncate at the base, aseptate, hyaline, guttulate, smooth-walled. Culture characteristics: Ascospores germinating on MEA within 24 h. Colonies growing on MEA, reaching 5 mm diam.

in 1 week at 28 °C, forming numerous water droplets on the mycelium. Mycelium superficial, slightly effuse, with undulate edge, ashy white, conidiomata produced in culture within 60 days. Material examined: THAILAND, Chiang Mai Province, Muang, Doi Su Thep, Huai Kok Ma, on dead wood, 21 April 2011, Saranyaphat Boonmee DST-04 (MFLU11-0135, holotype); ex-type living culture, MFLUCC 11-0376, BCC 52034. Lignosphaeria fusispora Boonmee, Thambugala & K.D. Hyde, sp. nov. Index Fungorum Number: IF551284; Facesoffungi number: FoF01091; Fig. 39 Etymology: Referring to the fusiform ascospores. Saprobic on dead wood. Sexual morph: Ascomata 326– 474μm high×248–292μm (x = 383×274μm, n=3) diam., scattered to gregarious, immersed, coriaceous to carbonaceous, black, subglobose, ostiolate. Ostiole central, rounded, periphysate. Peridium 40–70μm wide (x = 63×59μm, n=4), comprising several layers of brown to hyaline cells of textura angularis, fusing with the host tissue. Hamathecium comprising 1–2 μm wide, septate, unbranched, cellular pseudoparaphyses. Asci 47.5–74×8–13μm (x = 61×9μm,


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Fig. 38 Lignosphaeria thailandica (Asexual morph from the ex-type living culture). a. Germinating ascospore b, c. Colonies on MEA d. Vegetative hyphae in culture e, f. Asexual morph test and conidiomata developing on plant substrates g, h. Section of conidiomata and conidiophores developing in conidioma i, j. Conidiophores k. Conidiogenous cells with developing conidia l. Conidia. Scale bars: a, i–k=10μm, b, c, e= 10 mm, d, l=5μm, f=500μm, g, h=100μm

n=20), 8-spored, bitunicate, fissitunicate, cylindrical to clavate, pedicellate, apically rounded, with an indistinct ocular chamber. Ascospores 15–19×3–5μm (x = 17×4μm, n=10), bi to try-seriate, hyaline, narrow fusiform, wide at the second cell from the apex, narrow towards sub-acute ends, 3-septate, constricted at the septum, smooth-walled, lacking a mucilaginous sheath. Asexual morph: Undetermined. Notes: Lignosphaeria fusispora can be distinguished from L. thailandica, the type species of Lignosphaeria by having coriaceous to carbonaceous ascomata with rounded ostiole, 2–3-seriate, narrowly fusiform

ascospores with acute ends. In our phylogeny, L. fusispora clearly separates from L. thailandica and can be considered as a distinct species. Culture characteristics: Ascospores germinating on WA within 12 h. Colonies growing on MEA, reaching 5 mm diam. in 1 week at 28 °C, with numerous water droplets on mycelium. Mycelium superficial, slightly effuse, ashy white, sparsely hairy, with undulate edge. Material examined: THAILAND, Chiang Mai Province, Muang, Doi Su Thep, Huai Kok Ma, on dead wood, 21 April 2011, Saranyaphat Boonmee DST-05 (MFLU11–0136,

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Fig. 39 Lignosphaeria fusispora (holotype). a. Ascomata immersed on substrate b–d. Section of ascoma and peridium e. Pseudoparaphyses f– h. Asci i–l Ascospores m Germinating ascospore n, o. Colonies on MEA p. Asexual morph test but without developed conidiomata on plant substrates. Scale bars: a–c=100μm, d= 40μm, e=5μm, f–h=20μm, i– m=10μm, n–p=10 mm

holotype); ex-type living culture, MFLUCC 11-0377, BCC 52035.

Discussion In this paper we introduce 20 new genera which may appear excessive. However, we have only introduced new genera that have been resolved by molecular data and that have further support from morphological data. If we consider that only a small percentage of fungi have been discovered (Hawksworth 1991) and that tropical and saprobic fungi are under studied (Hyde et al. 2010), it is not surprising that there are large

numbers of undiscovered taxa. Furthermore, anything with a lophiostomataceous, crest-like ostiole was usually previously placed in Lophiostoma without considering other characters (Tanaka and Harada 2003a, Tanaka and Hosoya 2008, Hirayama et al. 2014, Liu et al. 2015). We have confidence in our newly introduced genera and predict that further collections of lophiostomataceous taxa will confirm our taxonomy and also reveal further new genera (Ariyawansa et al. 2015). Lophiostomataceae is a highly diverse family with many genera and species, while some species need to be recollected and sequenced as data are not available in GenBank. Morphologically they are highly diverse having different characters in ascomata, peridium, pseudoparaphyses and ascospores.


Molecular sequence data are important when identifying genera in Lophiostomataceae as there are several taxa morphologically similar to Lophiostoma (Tanaka and Harada 2003a, b, c; Tanaka et al. 2005b) which grouped in different genera and families (Mugambi and Huhndorf 2009; Hirayama et al. 2010; Hirayama and Tanaka 2011; Hyde et al. 2013). Lophiostoma (Pseudolophiostoma) vitigenum and L. fuckelii have been treated as Lophiotrema species because of their rather small ascomata and peridia of equal thickness (Tanaka and Harada 2003b). However, recent phylogenetic analysis as well as the present study revealed that these species belong to Lophiostomataceae (Hirayama and Tanaka 2011). Most strains in GenBank are named without attached voucher material and it is difficult to confirm their correct identification. Therefore, recollecting these fungus and epitypification with molecular sequence data are essential in order to ensure correct taxonomy. In this study, we separated Lophiostoma species into sixteen genera including Lophiostoma, Lophiohelichrysum, Dimorphiopsis, Platystomum and Vaginatispora, plus eleven newly introduced genera Biappendiculispora, Alpestrisphaeria, Capulatispora, Coelodictyosporium, Guttulispora, Lophiopoacea, Neotrematosphaeria, Paucispora, Pseudolophiostoma, Pseudoplatystomum and Sigarispora based on morphology and the results obtained from the phylogenetic analyses. However, we transferred only species having ex-type strains into the appropriate genus. The remaining species need to be confirmed with further sequence data from fresh collections. Mugambi and Huhndorf (2009) introduced Misturatosphaeria based on LSU and EF1-α sequence data. It is necessary to obtain additional sequence data (SSU, ITS) in order to obtain a better phylogenetic resolution for Floricolaceae. Mugambi and Huhndorf (2009) used an extraction kit to obtain DNA from whole fruiting bodies of Misturatosphaeria specimens And therefore other researches cannot study the culture characteristics and formation of asexual morphs of these taxa. Furthermore, Mugambi and Huhndorf (2009) have not illustrated morphological characters of their Misturatosphaeria species in detail. Therefore, recollection, illustration, isolation and molecular analysis of those Misturatosphaeria species are needed in order to derive a better understanding of these taxa. Wijayawardene et al. (2014a) synonymized Floricola and Misturatosphaeria under Macrodiplodiopsis Petr. based on phylogeny. In their phylogenetic analysis, a strain of Macrodiplodiopsis desmazieri (Mont.) Petr. clustered with Misturatosphaeria uniseriata. Wijayawardene et al. (2014b) listed four genera under the family Lophiostomataceae Sacc. (Dimorphiopsis Crous, Lophiostoma Ces. & De Not., Macrodiplodiopsis Petr. and Tumularia Descals & Marvanová). Based on new collection data, Macrodiplodiopsis desmazierii is clearly the asexual morphs of Splanchnonema platani (Ces.) M.E. Barr, and the fungus is apparently associated with a new family

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Macrodiplodiopsidaceae Voglmayr et al. (Crous et al. 2015). Therefore, in this study we did not follow the conclusions of Wijayawardene et al. (2014a) regarding the treatment of the genus Macrodiplodiopsis. However, we separated Misturatosphaeria species in to seven genera including Misturatosphaeria based on morphology and phylogeny. Angustimassarina the new genus we introduce in this study which is fungicolous and distributed on wide range of hosts. Putative strains of Massarina corticola (CBS 154.93) and Exosporium stylobatum (CBS 160.30) grouped in Angustimassarina clade and belong to this genus. However, M. corticola and E. stylobatum need to be recollected, sequenced and morphologycally studied in order to confirm the placement of these two species. Camarographium was introduced by Bubák (1916) and is typified by C. stephensii (Berk. & Broome) Bubák while currently Index Fungorum (2015) lists eight epithets. In our phylogenetic analysis Camarographium carpini Melnik et al. formed a separate clade between the Sporormiaceae clade and clade E. Another species C. koreanum Verkley et al. grouped in a separate lineage in Pleosporales. Camarographium carpini is not congeneric with C. koreanum, and recollections of the type species, C. stephensii, are required to resolve the familial placement (Crous et al. 2011). Acknowledgments The curators of the herbarium HHUF, IFRD and S are especially thanked for loaning herbarium specimens. Ying Zhang is thanked for providing EF sequence data for Misturatosphaeria mariae. We would like to thank The Mushroom Research Foundation, Chiang Rai, Thailand and Guizhou Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang, Guizhou Province, People’s Republic of China for providing postgraduate scholarship support to Kasun M. Thambugala. Kevin D. Hyde thanks the Chinese Academy of Sciences, project number 2013T2S0030, for the award of Visiting Professorship for Senior International Scientists at Kunming Institute of Botany. K. Tanaka would like to thank the Japan Society for the Promotion of Science (JSPS, 25440199 and 26291084) and Hirosaki University Grant for Exploratory Research by Young Scientists and Newly–appointed Scientists for financial support. Kasun M. Thambugala specially appreciates Profesor Jayarama Bhat and Dhanushka Udayanga for helpful comments and advice on the Manuscript. Shaun Pennycook is thanked for checking and suggesting corrections to most of the Latin names.

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