The Genera of Fungi – G6: Arthrographis, Kramasamuha, Melnikomyces, Thysanorea, and Verruconis

Abstract

The Genera of Fungi series, of which this is the sixth contribution, links type species of fungal genera to their morphology and DNA sequence data. Five genera of microfungi are treated in this study, with new species introduced in Arthrographis, Melnikomyces, and Verruconis. The genus Thysanorea is emended and two new species and nine combinations are proposed. Kramasamuha sibika, the type species of the genus, is provided with DNA sequence data for first time and shown to be a member of Helminthosphaeriaceae (Sordariomycetes). Aureoconidiella is introduced as a new genus representing a new lineage in the Dothideomycetes.

INTRODUCTION

This study focuses on five genera that form part of the Genera of Fungi project (www.generaoffungi.org; Crous et al. 2014a). The overall intention of this project is to revise and update the generic names of fungi, to provide DNA sequence data for them and to restudy or recollect their type species. In this study, we provide DNA sequence data for the unusual and poorly known genus Kramasamuha. Furthermore, the phylogenetic position of the genus Melnikomyces is clarified. We also resolve the taxonomy and phylogeny of Thysanorea and related Minimelanolocus species in the Herpotrichiellaceae. Additional new taxa are introduced based on morphological and DNA sequence data.

MATERIALS AND METHODS

Isolates

Freshly collected leaves and twigs were placed in damp chambers and treated as described by Castañeda-Ruiz et al. (2016). Protocols used for the collection and processing of soil samples are described in Giraldo et al. (2012, 2019) and Groenewald et al. (2018). After 1 wk of incubation on 2 % malt extract agar (MEA) supplemented with penicillin-G and streptomycin, individual colonies were transferred to MEA plates without antibiotics and incubated between 22–24 °C for 7–14 d, in order to obtain axenic cultures.

Colonies were sub-cultured onto 2 % potato dextrose agar (PDA), oatmeal agar (OA), MEA (Crous et al. 2019), autoclaved pine needles on 2 % tap water agar (PNA) (Smith et al. 1996), and incubated at 25 °C under continuous near-ultraviolet light to promote sporulation. Reference strains and specimens are maintained at the Westerdijk Fungal Biodiversity Institute (CBS Culture Collection and herbarium, respectively), Utrecht, The Netherlands or Coleção Octávio de Almeida Drummond (COAD), Viçosa, Brazil.

DNA isolation, amplification and analyses

Genomic DNA was extracted from fungal colonies growing on MEA using the Wizard^® Genomic DNA purification kit (Promega, Madison, WI), following the manufacturer’s protocol. The primers V9G (de Hoog & Gerrits van den Ende 1998) or ITS5 (White et al. 1990) and LR5 (Vilgalys & Hester 1990) were used to amplify part of the nuclear rDNA operon (ITS) spanning the 3′ end of the 18S nrRNA gene, the first internal transcribed spacer (ITS1), the 5.8S nrRNA gene, the second ITS region (ITS2) and approximately 900 bp of the 5′ end of the 28S nrRNA gene. The primers ITS4 (White et al. 1990) and LR0R (Vilgalys & Hester 1990) were used as internal sequence primers to ensure high quality sequences over the entire length of the amplicon. Part of the 18S small subunit nrRNA gene (SSU) was amplified and sequenced for selected isolates using NS1 and NS4 (White et al. 1990). Amplification conditions followed those described by Cheewangkoon et al. (2008). Part of the actin gene (act) was amplified and sequenced for selected isolates using the primer set Act1/Act4 (Voigt & Wöstemeyer 2000). The software SeqMan Pro v. 13.0.0 (DNASTAR, Madison, WI) and Geneious v. 11.0.4 (Kearse et al. 2012; https://www.geneious.com) were used to obtain consensus sequences of each isolate. Blast searches using ITS and LSU sequences were performed for each isolate and the closest matches were retrieved from GenBank and included in the phylogenetic analyses. Multiple sequence alignments for individual genes were generated using the online version of MAFFT (http://mafft.cbrc.jp/alignment/software/). Subsequent phylogenetic analyses from individual and combined datasets were conducted using Maximum-likelihood (ML) performed on the CIPRES Science Gateway portal (Miller et al. 2012) using RAxML v. 8.2.10 (Stamatakis 2014). The default parameters were used, and bootstrap support (BS) was obtained using the rapid bootstrapping algorithm with the automatic halt option. A BS ≥ 95 % was considered as statistically significant. Sequence data were deposited in the GenBank/ENA public databases (Table 1) and the alignments and trees in TreeBASE (http://www.treebase.org).

Table 1.

GenBank accession numbers of taxa included in this study.

Taxa	Isolates1	Country	Substrate	GenBank accession numbers2			References
				ITS	LSU	act/SSU
Dothideomycetes
Aliquandostipite khaoyaiensis	CBS 118232	Thailand	Twig	–	GU301796.1	–	Schoch et al. (2009)
	SS3028	Thailand		–	EF175649.1	–	Campbell et al. (2007)
Alternaria tenuissima	CBS 918.96	UK	Dianthus chinensis	–	KC584311.1	–	Woudenberg et al. (2013)
Alysidiella parasitica	CBS 120088	South Africa	Leaves Eucalyptus sp.	–	DQ923525.1	–	Summerell et al. (2006)
Apiosporina collinsii	CBS 118973	Canada	Amelanchier alnifolia	–	GU301798.1	–	Schoch et al. (2009)
Arthrographis arxii	CBS 203.78	India	Dung of herbivore	GQ272638.1	AB213426.1	HG316563.1/-	Murata et al. (2005), Kang et al. (2010), Giraldo et al. (2014a)
Arthrographis chlamydospora	CBS 135936	USA	Human urine	HG004554.1	HG004543.1	HG316560.1/-	Giraldo et al. (2014a)
Arthrographis curvata	CBS 135933	USA	Human nails	HG004557.1	HG004539.1	HG316557.1/-	Giraldo et al. (2014a)
	CBS 135934	Spain	River sediment	HG004556.1	HG004542.1	HG316558.1/-	Giraldo et al. (2014a)
Arthrographis globosa	UTHSC 11-757	USA	Bronchial wash	HG004553.1	HG004541.1	HG316561.1/-	Giraldo et al. (2014a)
Arthrographis grakistii	JW 22011 = CBS 145529	The Netherlands	Soil	MN794359	MN794336	MN816497	This study
	JW 22015	The Netherlands	Soil	MN794360	MN794337	MN816498	This study
	JW 22019	The Netherlands	Soil	MN794361	MN794338	MN816499	This study
	JW 49011	The Netherlands	Soil	MN794362	MN794339	MN816500	This study
	JW 49012	The Netherlands	Soil	MN794363	MN794340	MN816501	This study
	JW 180011	The Netherlands	Soil	MN794364	MN794341	MN816502	This study
	JW 190014= CBS 145530	The Netherlands	Soil	MN794365	MN794342	MN816503	This study
	JW 199018	The Netherlands	Soil	MN794366	MN794343	MN816504	This study
	JW 209002	The Netherlands	Soil	MN794367	MN794344	MN816505	This study
	JW 209003	The Netherlands	Soil	MN794368	MN794345	MN816506	This study
Arthrographis kalrae	CBS 693.77	India	Sputum	AB116536.1	AB116544.1	HG316544.1/-	Xi et al. (2004), Giraldo et al. (2014a)
	JW 21004	The Netherlands	Soil	MN794369	MN794346	MN816507	This study
	JW 21008 = CBS 145527	The Netherlands	Soil	MN794370	MN794347	MN816508	This study
	JW 21029	The Netherlands	Soil	MN794371	MN794348	MN816509	This study
Arthrographis longispora	CBS 135935	USA	Human foot	HG004555.1	HG004540.1	HG316559.1/-	Giraldo et al. (2014a)
	JW 22007 = CBS 145528	The Netherlands	Soil	MN794372	MN794349	MN816510	This study
Asterina chrysophylli	VIC 42823	Brazil	Leaves Henriettea succosa	–	KP143738.1	–	Guatimosim et al. (2015)
Asterina melastomatis	VIC 42822	Brazil	Leaves Miconia sp.	–	NG_057055.1	–	Guatimosim et al. (2015)
Asterotexis cucurbitacearum	PMA M141224	Panama	Sechium edule	–	HQ610510.1	–	Unpublished
	VIC 24814	Brazil	Leaves Cucurbita pepo	–	NG_057054.1	–	Guatimosim et al. (2015)
Aulographina eucalypti	CPC 12986	Australia	Eucalyptus cloeziana	–	HM535600.1	–	Cheewangkoon et al. (2012)
Aureoconidiella foliicola	CBS 145943	South Africa	Leaves Syzygium cordatum	MN794373	MN794350	–	This study
Bezerromyces brasiliensis	URM7411	Brazil	Tacinga inamoena	–	KX518623.1	–	Bezerra et al. (2017)
Bezerromyces pernambucoensis	URM7412	Brazil	Tacinga inamoena	–	KX518624.1	–	Bezerra et al. (2017)
Blastacervulus eucalypti	CBS 124759	Australia	Eucalyptus robertsonii subsp. hemisphaerica	–	GQ303302.1	–	Cheewangkoon et al. (2009)
Blastacervulus robbenensis	CBS 124780	Cyprus	Eucalyptus sp.	–	HM628777.1	–	Cheewangkoon et al. (2012)
Botryosphaeria dothidea	CBS 115476	Switzerland	Prunus sp.	–	DQ377852.1	–	Crous et al. (2006b)
Brachiosphaera tropicalis	E192	–	–	–	EF175653.1	–	Campbell et al. (2007)
Byssosphaeria jamaicana	SMH 1403	–	–	–	GU385152.1	–	Mugambi & Huhndorf (2009a)
Byssosphaeria salebrosa	SMH 2387	–	–	–	GU385162.1	–	Mugambi & Huhndorf (2009a)
Capnodium coffeae	CBS 147.52	Zaire	Berry Coffea robusta	–	MH868489.1	–	Vu et al. (2019)
Cladoriella eucalypti	CBS 115899	South Africa	Leaves Eucalyptus	–	EU040224.1	–	Crous et al. (2007b)
Cladoriella kinglakensis	CPC 32730	Australia	Leaves Eucalyptus	–	MG386126.1	–	Crous et al. (2017)
Cladoriella paleospora	CBS 124761	Australia	Leaves Eucalyptus	–	GQ303303.1	–	Cheewangkoon et al. (2009)
Cladoriella rubrigena	CBS 124760	Australia	Leaves Eucalyptus	–	MH874921.1	–	Vu et al. (2019)
Cladoriella xanthorrhoeae	CBS 143398	Australia	Leaves Xanthorrhoea sp.	–	NG_059054.1	–	Crous et al. (2017)
Cladosporium halotolerans	CBS 127371	Cuba	Human	–	MH875988.1	–	Vu et al. (2019)
Cladosporium variabile	CBS 121636	USA	Spinacia oleracea	–	MH874684.1	–	Vu et al. (2019)
Clavatispora thailandica	MFLUCC 17-2237	Thailand	Hevea brasiliensi	–	MH062960.1	–	Unpublished
	MFLUCC 10-0107	Thailand	Dead stems	–	NG_058863.1	–	Boonmee et al. (2014)
Dibotryon morbosum	N/A	USA	Prunus sp.	–	EF114694.1	–	Winton et al. (2007)
Diplodia mutila	CBS 431.82	The Netherlands	Dead branches Fraxinus excelsior	–	DQ377863.1	–	Crous et al. (2006b)
Dissoconium aciculare	CBS 204.89	Germany	Astragalus	–	GU214419.1	–	Crous et al. (2009a)
Eremomyces bilateralis	CBS 781.70	USA	Dung of pack rat	HG004552.1	HG004545.1	HG316562.1/-	Giraldo et al. (2014a)
Fusicladium pini	CBS 463.82	The Netherlands	Needle Pinus sylvestris	–	EU035436.1	–	Crous et al. (2007c)
Fusicladium ramoconidii	CBS 462.82	The Netherlands	Needle Pinus sp.	–	EU035439.1	–	Crous et al. (2007c)
Gibbera conferta	CBS 191.53	Switzerland	Vaccinium uliginosum	–	GU301814.1	–	Schoch et al. (2009)
Gloniopsis arciformis	GKM L166A	–	–	–	GU323211.1	–	Schoch et al. (2009)
Glonium circumserpens	CBS 123342	Tasmania	Wood	–	FJ161208.1	–	Boehm et al. (2009)
Glonium circumserpens	CBS 123343	Tasmania	Saxicolous on limestone	–	FJ161200.1	–	Boehm et al. (2009)
Helicomyces roseus	CBS 283.51	Switzerland	Dead bark	–	AY856881.1	–	Tsui et al. (2006)
Herpotrichia juniperi	AFTOL-ID 1608	Switzerland	Juniperus nana	–	DQ678080.1	–	Schoch et al. (2009)
Heteroconium eucalypti	CBS 120122	Uruguay	Leaves Eucalyptus dunnii	–	DQ885893.1	–	Crous et al. (2006a)
Hysterium angustatum	CBS 123334	USA	Bark Pinus rigida	–	FJ161207.1	–	Boehm et al. (2009)
Hysterium pulicare	ANM1455	USA	–	–	GQ221904.1	–	Mugambi & Huhndorf (2009b)
Hysteropatella clavispora	CBS 247.34	USA	Salix sp.	–	AY541483.1	–	Lumbsch et al. (2005)
Hysteropatella prostii	H.B. 9934b	Germany	Malus domestica	–	KT876980.1	–	Unpublished
Jahnula appendiculata	SS2900	Thailand	–	–	EF175654.1	–	Campbell et al. (2007)
Jahnula aquatica	R68-1	USA	–	–	EF175655.1	–	Campbell et al. (2007)
Jahnula bipileata	F49-1	USA	–	–	EF175657.1	–	Campbell et al. (2007)
Jahnula seychellensis	SS2113.1	Thailand	–	–	EF175665.1	–	Campbell et al. (2007)
Leptoxyphium fumago	CBS 123.26	Indonesia	Hibiscus tiliaceus	–	GU214430.1	–	Crous et al. (2009a)
Macrophomina phaseolina	CBS 227.33	–	Zea mays	–	DQ377906.1	–	Crous et al. (2006b)
Melanomma pulvis-pyrius	CBS 124080	France	Bark Salix caprea	–	GU456323.1	–	Zhang et al. (2009a)
Melnikomyces thailandicus	CBS 145767	Thailand	Soil	MN794374	MN794351	–	This study
Melnikomyces vietnamensis	CBS 136209	Vietnam	Leaves	–	NG_058087.1	–	Crous et al. (2014b)
Mycosphaerella punctiformis	CBS 113265	The Netherlands	Dead leaves Quercus robur	–	DQ470968.1	–	Spatafora et al. (2006)
Neocoleroa metrosideri	PDD107531	New Zealand	Metrosideros excelsa	–	NG_059638.1	–	Johnston & Park (2016)
Neofusicoccum mangiferae	CBS 118532	Australia	Mangifera indica	–	NG_055730.1	–	Crous et al. (2006b)
Neofusicoccum nonquaesitum	CBS 126655	USA	Umbellularia californica	–	NG_058258.1	–	Yang et al. (2017)
Ochroconis constricta	CBS 202.27	USA	Soil	–	KF156147.1	–	Samerpitak et al. (2014)
Ochroconis gamsii	CBS 239.78	Sri Lanka	Leaf Caryota plumosa	–	NG057992.1	–	Samerpitak et al. (2014)
Patellaria cf. atrata	BCC 28876	Thailand	–	–	GU371828.1	–	Schoch et al. (2009)
	BCC 28877	Thailand	–	–	GU371829.1	–	Schoch et al. (2009)
Phaeocryptopus gaeumannii	CBS 267.37	Germany	Pseudotsuga menziesii	–	EF114698.1	–	Winton et al. (2007)
Phaeotrichum benjaminii	CBS 541.72	–	Dung of rodent	–	AY779311.1	–	Lumbsch et al. (2005)
Phoma herbarum	CBS 567.63	USA	Fruit Malus sylvestris	–	MH869982.1	–	Vu et al. (2019)
Pirozynskiella laurisilvatica	CBS 138109	Spain	Leaves Laurus sp.	–	NG_058462.1	–	Hernández-Restrepo et al. (2017)
Psiloglonium simulans	CBS 206.34	USA	Tilia sp.	–	FJ161178.1	–	Boehm et al. (2009)
Rhexothecium globosum	CBS 955.73	Egypt	Desert soil	MH860827.1	HG004544.1	–	Giraldo et al. (2014a), Vu et al. (2019)
Rhytidhysteron rufulum	CBS 306.38	–	Pistacia chinensis	–	FJ469672.1	–	Schoch et al. (2009)
Schizothyrium pomi	CBS 228.57	Italy	–	–	EF134947.1	–	Batzer et al. (2008)
	CBS 486.50	The Netherlands	Polygonum sachalinense	–	EF134948.1	–	Batzer et al. (2008)
Scolecobasidiella avellanea	CBS 772.73	Somalia	Soil	–	EF204505.1	–	Unpublished
Stemphylium herbarum	CBS 191.86	India	Leaf Medicago sativa	–	JX681120.1	–	Verkley et al. (2014)
Sympoventuria capensis	CBS 120136	South Africa	Leaf litter Eucalyptus sp.	–	NG_057984.1	–	Samerpitak et al. (2014)
Teratosphaeria destructans	CBS 111369	Indonesia	Eucalyptus grandis	–	EU019287.2	–	Crous et al. (2007a)
Teratosphaeria fibrillosa	CBS 121707	South Africa	Leaves Protea sp.	–	KF902075.1	–	Quaedvlieg et al. (2014)
Teratosphaeria stellenboschiana	CBS 116428	South Africa	Leaf litter Eucalyptus sp.	–	EU019295.1	–	Crous et al. (2007a)
Trichodelitschia bisporula	CBS 262.69	The Netherlands	Dung of Rabbit	–	GU348996.2	–	Schoch et al. (2009)
Tubeufia paludosa	CBS 245.49	The Netherlands	Corylus avellana	–	MH856510.1	–	Vu et al. (2019)
Uwebraunia commune	CBS 110747	South Africa	Eucalyptus nitens	–	GQ852589.1	–	Crous et al. (2009b)
Uwebraunia dekkeri	CBS 111282	Zambia	Eucalyptus globulus	–	GU214425.1	–	Crous et al. (2009b)
Venturia inaequalis	CBS 176.42	France	–	–	GU348998.1	–	Schoch et al. (2009)
Venturia populina	CBS 256.38	Italy	Populus canadensis	–	GU323212.1	–	Schoch et al. (2009)
Verruconis calidifluminalis	CBS 125818	Japan	Hot spring effluent	–	NG_057985.1	–	Samerpitak et al. (2014)
Verruconis gallopava	CBS 547.81	New Zealand	–	–	KF156109.1	–	Samerpitak et al. (2014)
	CBS 437.64	USA	Brain abscess Meleagris gallopavo	–	NG_58016.1	–	Machouart et al. (2014)
Verruconis thailandica	CBS 145768	Thailand	Soil	MN794375	MN794352	–	This study
Verruconis verruculosa	CBS 119775	Malaysia	Root Hevea species	–	KF282668.1	–	Machouart et al. (2014)
Xiliomyces brasiliensis	URM7413	Brazil	Tacinga inamoena	–	KX518625.1	–	Bezerra et al. (2017)
Zasmidium cellare	CBS 146.36	–	Wall in wine cellar	–	EU041878.1	–	Arzanlou et al. (2007)
Eurotiomycetes
Aculeata aquatica	MFLUCC 11-0529	Thailand	Submerged wood	MG922575.1	MG922579.1	–/MG922571.1	Dong et al. (2018)
Capronia pilosella	AFTOL-ID 657	–	–	DQ823099.1	DQ823106.1	–/DQ826737.1	James et al. (2006)
Cladophialophora carrionii	CBS 160.54	Australia	Man	FJ358234.1	FJ358302.1	–/AF050262.1	Gueidan et al. (2008), Untereiner & Naveau (1999)
Cladophialophora minourae	CBS 556.83	Japan	Decaying wood	FJ358235.1	FJ358303.1	–/AY251087.1	Braun et al. (2003), Gueidan et al. (2008)
Cladophialophora parmeliae	CBS 129337	Portugal	–	JQ342182.1	–	–/JQ342180.1	Diederich et al. (2013)
Cladophialophora subtilis	CBS 122642	The Netherlands	Ice tea	NG_058961.1	KX822283.1	–/NR_111363.1	Badali et al. (2008), Vasse et al. (2017)
Cyphellophora oxyspora	CBS 698.73	Sri Lanka	–	KC455262.1	KC455305.1	–/KC455249.1	Réblová et al. (2013)
Cyphellophora sessilis	CBS 243.85	The Netherlands	Resin Picea abies	EU514700.1	KC455308.1	–/EU514700.1	Untereiner et al. (2008), Réblová et al. (2013)
Exophiala jeanselmei	CBS 507.90	Uruguay	Man	FJ358242.1	FJ358310.1	–/NR_111129.1	Gueidan et al. (2008)
Exophiala nigra	dH 12,296	–	–	FJ358244.1	FJ358312.1	–	Gueidan et al. (2008)
Exophiala pisciphila	CBS 537.73	USA	Ictalurus punctatus	MH872483.1	JN856018.1	–/AF050272.1	de Hoog et al. (2011), Untereiner & Naveau (1999), Vu et al. (2019)
	AFTOL-ID 669	–	–	DQ823101.1	DQ823108.1	–/DQ826739.1	Gueidan et al. (2008)
Exophiala salmonis	AFTOL-ID 671	–	–	EF413609.1	EF413608.1	–	Geiser et al. (2006)
	CBS 157.67	Canada	Salmo clarkii	MH870616.1	JN856020.1	–/NR_121270.1	de Hoog et al. (2011), Schoch et al. (2014), Vu et al. (2019)
Exophiala xenobiotica	CBS 115831	Germany	Browncol	FJ358246.1	FJ358314.1	–/AY857539.1	Gueidan et al. (2008)
Fonsecaea monophora	CBS 102243			FJ358247.1	FJ358315.1	–/EU938579.1	Gueidan et al. (2008)
Melanoctona tectonae	MFLUCC 12-0389	Thailand	Tectona grandis	KX258779.1	KX258780.1	–/KX258778.1	Unpublished
Phialophora americana	AFTOL-ID 658	–	–	FJ358226.1	FJ358294.1	–	Gueidan et al. (2008)
Phialophora verrucosa	AFTOL-ID 670	–	–	EF413615.1	EF413614.1	–	Geiser et al. (2006)
Rhinocladiella anceps	AFTOL-ID 659	–	–	DQ823102.1	DQ823109.1	–/DQ826740.1	James et al. (2006)
Rhinocladiella anceps	CBS 181.65	Canada	Soil	EU041862.1	AY554292.1	–/MH858534.1	Arzanlou et al. (2007)
Thysanorea asiatica	MFLUCC 15-0237	China	Submerged wood	KR215610.1	KR215615.1	–/KR215604.1	Liu et al. (2015)
Thysanorea cantrelliae	CBS 145909	USA	Unidentified twig	MN794376	MN794353	–/MN794382	This study
Thysanorea curvata	MFLUCC 15-0259	China	Submerged wood	KR215609.1	KR215614.1	–/KR215605.1	Liu et al. (2015)
Thysanorea lotorum	CBS 235.78	USA	Root Lotus corniculatus	MH872892.1	–	–/MH861130.1	Vu et al. (2019)
	KUMCC 15-0206	China	Submerged wood	KX789215.1	–	–/KX789212.1	Liu et al. (2015)
Thysanorea melanica	MFLUCC 15-0415	China	Submerged wood	KR215613.1	KR215618.1	–/KR215608.1	Liu et al. (2015)
Thysanorea nonramosa	MFLUCC 17-2378	Thailand	Wood	MH532970.1	–	–/MH532971.1	Wang et al. (2019)
Thysanorea obscura	MFLUCC 15-0416	China	Submerged wood	KR215611.1	KR215616.1	–/KR215606.1	Liu et al. (2015)
Thysanorea papuana	CBS 212.96	Papua New Guinea	–	EU041871.1	–	–/EU041814.1	Arzanlou et al. (2007)
	MFLUCC 15-0966	Thailand	Submerged wood	MG922576.1	MG922580.1	–/MG922572.1	Dong et al. (2018)
Thysanorea rousseliana	CBS 126086	Spain	Dead branches Quercus ilex	MH875246.1	–	–/MH863784.1	Vu et al. (2019)
Thysanorea seifertii	CBS 145910	USA	Unidentified twig	MN794377	MN794354	–/MN794383	This study
Thysanorea thailandensis	MFLUCC 15-0971	Thailand	Submerged wood	MG922577.1	MG922581.1	–/MG922573.1	Dong et al. (2018)
Thysanorea yunnanense	MFLUCC 15-0414	Thailand	Submerged wood	KR215612.1	KR215617.1	–/KR215607.1	Liu et al. (2015)
Veronaea botryosa	CBS 254.57	Italy	Sansa olive	EU041873.1	JN856021.1	–/EU041816.1	Arzanlou et al. (2007)
	MFLUCC 11-0072	Thailand	Submerged wood	MG922574.1	MG922578.1	–/MG922570.1	Dong et al. (2018)
Veronaea compacta	CBS 268.75	South Africa	–	EU041876.1	–	–/EU041819.1	Arzanlou et al. (2007)
Veronaea japonica	CBS 776.83	Japan	Dead bamboo culm	EU041875.1	–	–/EU041818.1	Arzanlou et al. (2007)
Sordariomycetes
Anthostomella sp.	SMH3101	USA	–	–	AY780050.1	–	Miller & Huhndorf (2005)
Camarops tubulina	SMH4614	Denmark	–	–	AY346266.1	–	Huhndorf et al. (2004)
Camarops ustulinoides	SMH1988	USA	–	–	AY346267.1	–	Huhndorf et al. (2004)
Chaetosphaeria ovoidea	SMH2605	USA	–	–	AF064641.1	–	Fernandez et al. (1999)
Coniochaeta discoidea	SANK 12878	–	–	–	AY346297.1	–	Huhndorf et al. (2004)
Coniochaetidium savoryi	TRTC 51980	–	–	–	AY346276.1	–	Huhndorf et al. (2004)
Cytospora ceratosperma	AR3426	Austria	Quercus robur	–	AF408387.1	–	Castlebury et al. (2002)
Diaporthe phaseolorum	FAU458	USA	–	–	AY346279.1	–	Huhndorf et al. (2004)
Echinosphaeria canescens	JHC97-006	–	–	–	KF765604.1	–	Miller et al. (2014)
	SMH4666	–	–	–	KF765605.1	–	Miller et al. (2014)
	SMH4791	–	–	–	AY436403.1	–	Miller & Huhndorf (2004)
	TL5730	–	–	–	AY436404.1	–	Miller & Huhndorf (2004)
Eutypa sp.	SMH3580	Panama	Branch	–	AY346280.1	–	Huhndorf et al. (2004)
Fusarium ambrosium	SMH1999	–	–	–	AY780077.1	–	Miller & Huhndorf (2005)
Helminthosphaeria carpathica	SMH3903	–	–	–	KF765606.1	–	Miller et al. (2014)
Helminthosphaeria cf. stuppea	JF04120	–	–	–	KF765611.1	–	Miller et al. (2014)
	TL11998	–	–	–	KF765612.1	–	Miller et al. (2014)
Helminthosphaeria clavariarum	SMH4609	Denmark	Clavulina cristata	–	AY346283.1	–	Huhndorf et al. (2004)
Helminthosphaeria corticiorum	JF04225	–	–	–	KF765607.1	–	Miller et al. (2014)
Helminthosphaeria odontiae	ANM928	–	–	–	KF765610.1	–	Miller et al. (2014)
Helminthosphaeria tomaculum	SMH2485	–	–	–	KF765613.1	–	Miller et al. (2014)
Helminthosphaeria triseptata	JF04015	–	–	–	KF765614.1	–	Miller et al. (2014)
Hilberina caudata	SMH1542	–	–	–	KF765615.1	–	Miller et al. (2014)
Hilberina munkii	SMH1531	–	–	–	KF765616.1	–	Miller et al. (2014)
Kramasamuha sibiki	CPC 35619 = CBS 146338	Australia	Leaves Lophostemon confertus	MN794378	MN794355	–	This study
	CPC 36725 = CBS 146339	Malaysia	Needles Pinus tecunumanii	MN794379	MN794356	–	This study
	CBS 146133 = CPC 36153	South Africa	Leaves Syzygium cordatum	MN794380	MN794357	–	This study
	COAD 2632	Brazil	Leaves Hypericum innodorum	MN794381	MN794358	–	This study
Lasiosphaeria ovina	SMH1538	–	–	–	AF064643.1	–	Fernandez et al. (1999)
Neurospora crassa	MUCL 19026	–	–	–	AF286411.1	–	Untereiner et al. (2001)
Ruzenia spermoides	ANM163	–	–	–	KF765618.1	–	Miller et al. (2014)
	SMH4606	–	–	–	AY436422.1	–	Miller & Huhndorf (2004)
	SMH4655	–	–	–	KF765619.1	–	Miller et al. (2014)
Sporoschisma hemipsila	SMH2125	–	–	–	AY346292.1	–	Huhndorf et al. (2004)
Synaptospora plumbea	ANM963	–	–	–	KF765620.1	–	Miller et al. (2014)
	SMH3962	–	–	–	KF765621.1	–	Miller et al. (2014)
Valsonectria pulchella	SMH1193	–	–	–	AY346304.1	–	Huhndorf et al. (2004)

¹ BCC: BIOTEC Culture Collection, National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok, Thailand; CBS: Culture Collection of the Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands; COAD: Coleção Octávio de Almeida Drummond, Viçosa, Brazil; CPC: Culture Collection of Pedro Crous, Utrecht, The Netherlands; JW: Johanna Westerdijk Culture Collection, Utrecht, The Netherlands; MFLUCC: Mae Fah Luang University Culture Collection, Chiang Rai, Thailand; UTHSC: Fungus Testing Laboratory of the University of Texas Health Science Center at San Antonio, USA. For other acronyms see references.

² LSU: Large subunit of the nrDNA; SSU: Small subunit of the nrDNA; ITS: internal transcribed spacer regions of the nrDNA and intervening 5.8S nrDNA; act: partial actin gene. Accession numbers of sequences newly generated in this study are indicated in bold.

Morphology

Slide preparations were mounted in lactic acid or water from colonies sporulating on the media previously mentioned. Observations were made with a Nikon SMZ1500 dissecting microscope and with a Nikon Eclipse Ni compound microscope using a DSRi2 digital camera (Nikon, Tokyo, Japan) and NIS-Elements imaging software v. 4.3. Colony characters and pigment production were noted after 1–2 wk of growth on MEA, PDA and OA incubated at 25 ºC under natural light. Colony colours (surface and reverse) were determined using the colour charts of Rayner (1970). Taxonomic novelties were deposited in MycoBank (www.MycoBank.org; Crous et al. 2004).

RESULTS

Phylogeny

Three overview phylogenies were generated in this study. The first two of these were based on a partial alignment of LSU to provide the phylogenetic position of the treated genera and species within the Dothideomycetes (Fig. 1) and Sordariomycetes (Fig. 2). A third analysis was implemented for selected Herpotrichiellaceae genera based on a concatenated ITS/LSU/SSU alignment (Fig. 3). Other phylogenetic trees specific to the treated species are discussed in the notes for those taxa.

Fig. 1.

Maximum composite likelihood tree obtained from the RAxML analysis of the LSU sequence alignment of selected Dothideomycetes. Bootstrap support values above 70 % are shown at the nodes. Families and orders are indicated with coloured blocks to the right of the tree. Taxonomic novelties described in this study are indicated in boldface. The tree was rooted to Yarrowia hollandica (CBS 4855).

Fig. 2.

Maximum composite likelihood tree obtained from the RAxML analysis of the LSU sequence alignment of selected Sordariomycetes. Bootstrap support values above 70 % are shown at the nodes. Families and orders are indicated with coloured blocks to the right of the tree. Included strains described in this study are indicated in boldface. The tree was rooted to Orbilia vinosa (AFTOL-ID 905) and Orbilia auricolor (AFTOL-ID 906).

Fig. 3.

Maximum composite likelihood tree obtained from the RAxML analysis of the ITS/LSU/SSU sequence alignment of selected Herpotrichiellaceae. Bootstrap support values above 70 % are shown at the nodes. Taxonomic novelties described in this study are indicated in boldface. The tree was rooted to Cyphellophora oxyspora (CBS 698.73) and Cyphellophora sessilis (CBS 243.85).

The BLAST search results using the LSU and ITS sequences for the isolates CBS 145943, CBS 145767, CBS 145768, and JW showed that they were related to members of Cladoriellales and Asterinales, Venturiales and Eremomycetales in the Dothideomycetes. Consequently, the currently accepted taxa in those orders and other orders in the Dothideomycetes were included in our analyses (Fig. 1).

The overview phylogeny of Dothideomycetes (Fig. 1) revealed that in the Arthrographis subclade (96 % BS), the strains JW 49012, JW 190014, and JW 209002 grouped together in a separate clade representing a putative new species that is described below.

The genus Aureoconidiella is introduced to accommodate CBS 145943 that formed a lineage distinct from other genera, families and orders included in the analysis. A new family and order are introduced for this genus. Furthermore, the isolates CBS 145767 and CBS 145768 nested in the Sympoventuriaceae clade (91 % BS) within the Venturiales. They were closely related but different to Melnikomyces vietnamensis and Verruconis verruculosa, respectively. These isolates are consequently considered to represent putative new species in the genera Melnikomyces and Verruconis that are introduced below.

The overview phylogeny of Sordariomycetes (Fig. 2) revealed that four isolates, CBS 146133, CBS 146338, CBS 146339, and COAD 2632 of Kramasamuha sibika grouped together in a fully-supported terminal clade (100 % BS), related to Helminthosphaeriaceae (97 % BS).

The combined analysis of the ITS/LSU/SSU (Fig. 3) revealed that Minimelanolocus and Thysanorea cluster together in the same clade (86 % BS). Several species treated so far as belonging to Minimelanolocus, i.e. M. aquaticus, M. asiaticus, M. curvatus, M. melanicus, M. obscurus, M. rosselianus, M. submersus, and M. thailandensis, proved to be congeneric with Thysanorea and therefore new combinations are proposed to accommodate them. In addition, CBS 145909 and CBS 145910 formed a separate subclade (100 % BS) within Thysanorea, representing two putative new species.

Taxonomy

Aureoconidiellales Hern.-Restr. & Crous, ord. nov. MycoBank MB833918.

Description: See description of Aureoconidiella.

Type family: Aureoconidiellaceae Hern.-Restr. & Crous

Aureoconidiellaceae Hern.-Restr. & Crous, fam. nov. MycoBank MB833917.

Description: See description of Aureoconidiella.

Type genus: Aureoconidiella Hern.-Restr. & Crous

Aureoconidiella Hern.-Restr. & Crous, gen. nov. MycoBank MB833915.

Etymology: Name refers to the golden brown colour of its conidia.

Conidiophores macronematous, simple, septate, brown. Conidiogenous cells integrated, terminal, polyblastic, with thickened scars, brown to pale brown. Conidia globose to subglobose with apiculate base, initially subhyaline, golden brown at maturity, verrucose.

Type species: Aureoconidiella foliicola Hern.-Restr. & Crous

Aureoconidiella foliicola Hern.-Restr. & Crous, sp. nov. MycoBank MB833916. Fig. 4.

Fig. 4.

Aureoconidiella foliicola gen. et sp. nov. (CBS 145943). A. Conidiophores and conidia. B. Conidiophore with conidiogenous cell. C, D. Conidiogenous cells giving rise to conidia. E. Conidia. Scale bars: A = 20 μm, all others = 10 μm.

Etymology: The epithet “foliicola” refers to its habitat on a dead leaf.

Mycelium consisting of septate, smooth, brown, 1–2.5 μm wide hyphae. Conidiophores macronematous, simple, septate, brown, 40–85 × 3–5.5 μm. Conidiogenous cells integrated, terminal, polyblastic, with thickened scars, brown to pale brown, 25–53 × 3–4.5 μm. Conidia globose to subglobose with apiculate base, initially subhyaline, golden brown at maturity, verrucose, 5–8 μm diam, base 1–2 μm wide.

Culture characteristics: On MEA and OA surface cottony aerial mycelium Fawn, sepia to dark brick close to the agar, margin effuse, entire; reverse sepia to black.

Typus: South Africa, KwaZulu-Natal, Richards Bay, on living leaves of Syzygium cordatum (Myrtaceae), Jun. 2016, M.J. Wingfield (holotype CBS H-24099, culture ex-type CPC 36154 = CBS 145943).

Notes: This new lineage is introduced to accommodate a fungus characterised by unbranched conidiophores, cicatrised and sympodial conidiogenous cells with thickened scars, producing sub-globose, verruculose, and golden brown conidia. Other related lineages are those accommodating Asterinales and Cladoriellales (Fig. 1). However, they differ from those in the Aureoconidiellales based on the morphology of the asexual morphs. The Asterinales is mainly characterised by taxa that are coelomycetes with pycnothyrial conidiomata (Guatimosim et al. 2015, Jaklitsch et al. 2016). The Cladoriellales is a monotypic order related to cladosporium-like hyphomycetous fungi with conidia frequently remaining attached in long acropetal chains (Crous et al. 2006c, 2017).

Authors: M. Hernández-Restrepo, P.W. Crous and M.J. Wingfield

Arthrographis Sigler & J.W. Carmich., Mycotaxon 4: 359. 1976.

Synonym: [Arthrographis G. Cochet, Annls Parasit. Hum. Comp. 17: 97. 1939. (Nom. inval., Art. 39.1)]

Vegetative hyphae septate, hyaline, smooth- and thin-walled. Conidiophores macro- or micronematous, erect, simple or poorly branched, hyaline, smooth-walled. Conidiogenous hyphae simple or branched, thin-walled, forming septa basipetally to form arthroconidia released by schizolytic secession. Arthroconidia unicellular, cylindrical or cuboid, straight, subhyaline, thick- and smooth-walled. Synasexual morph trichosporiella-like, with conidia growing directly on undifferentiated hyphae, sessile, lateral, terminal, globose, subglobose or clavate, subhyaline, thin- and smooth-walled. Sexual morph not observed.

Type species: Arthrographis kalrae (R.P. Tewari & Macph.) Sigler & J.W. Carmich.

Arthrographis kalrae (R.P. Tewari & Macph.) Sigler & J.W. Carmich., Mycotaxon 4: 360. 1976.

Basionym: Oidiodendron kalrae R.P. Tewari & Macph., Mycologia 63: 603. 1971 [as ‘kalrai’].

Synonym: [Arthrographis langeronii G. Cochet (as ‘langeroni’), Annls Parasit. hum. comp. 17: 97. 1939. (Nom. inval., Art. 39.1)]

Descriptions and illustrations: Sigler & Carmichael (1976, 1983), Giraldo et al. (2014a).

Specimens examined: The Netherlands, Utrecht, isolated from soil, 2017, E. Kieviet (JW 21004 = CBS 145527), ibid. JW 21008, ibid. JW 21029.

Arthrographis grakistii Giraldo López & Hern.-Restr., sp. nov. MycoBank MB833677. Fig. 5.

Fig. 5.

Arthrographis grakistii sp. nov. (CBS 145530). A. Poorly branched conidiophores and arthroconidia. B, C. Trichosporiella-like synasexual morph. Scale bars = 10 μm.

Etymology. Named after Ewan Grakist, who collected the soil sample. This species was discovered as part of a Citizen Science project in the Netherlands.

Vegetative hyphae septate, hyaline, smooth- and thin-walled, 1.5–2 μm wide. Conidiophores semi-macronematous or micronematous, erect, simple, hyaline, smooth-walled. Conidiogenous hyphae simple or branched, 1.5–2 μm wide, thin-walled, forming septa basipetally to form arthroconidia released by schizolytic secession. Arthroconidia unicellular, cylindrical or cuboid, straight, 2–4 × 2–3 μm, subhyaline, thick- and smooth-walled. Synasexual morph trichosporiella-like with conidia growing directly on undifferentiated hyphae, sessile, sometimes with a subcylindrical to clavate intercalary cell, lateral, terminal, globose, subglobose or clavate, 3–5 × 2–3 μm, subhyaline, thin- and smooth-walled. Sexual morph not observed.

Culture characteristics: Colonies at 25 °C after 14 d: on OA reaching 13–14 mm, flat, glabrous to floccose, surface and reverse buff. On MEA and PDA reaching 11–13 mm and 12–17 mm, respectively; flat or raised, dusty to cottony at centre, glabrous toward the periphery, buff to honey, reverse uncoloured. No growth at 37 °C.

Typus: The Netherlands, Utrecht Province, Wijk bij Duurstede, from garden soil, 2017, E. Grakist (holotype CBS H-23912, culture ex-type CBS 145530 = JW 190014).

Additional materials examined: The Netherlands, Utrecht Province, IJsselstein, from garden soil, 2017, J. Brus (JW 209002, JW 209003); from garden soil, 2017, R. de Bruyn (JW 180011); Utrecht, from garden soil, 2017, M. Wickham (JW 199018); Zeeland Province, Vlissingen, from garden soil, 2017, N. Penabad (CBS 145529 = JW 22011, JW 22015, JW 22019); Gelderland Province, Zaltbommel, from garden soil, 2017, K. & T. de Man (JW 49011, JW 49012).

Notes: Based on a BLAST search using the ITS and LSU loci several soil isolates (JW isolates listed in Table 1) were identified as belonging to Arthrographis. In order to confirm their identity at the species level, a combined analysis of the LSU/ITS/act loci was performed, including all members from Eremomycetaceae. The ML tree (Fig. 6) showed that three isolates (JW 21004, JW 21008 and JW 21029) grouped with the type species of A. kalrae (CBS 693.77) and one (JW 22007) with A. longispora (CBS 135935). However, most of the isolates formed a well-supported clade that represents the new species A. grakistii.

Fig. 6.

Maximum composite likelihood tree based on partial sequences from the LSU, ITS and act regions from all members of Eremomycetales. Colour boxes (blue and green) indicate the generic clades. Bootstrap support values above 70 % are shown at the nodes. Ex-type strains are in boldface. T = Ex-type.

The phylogenetic analyses showed that A. grakistii is closely related to A. longispora (Figs 1, 6). The latter species, however, has longer and narrower arthroconidia [5–10(–13) × 1–1.5 μm in A. longispora vs. 2–4 × 2–3 μm in A. grakistii] and does not produce the trichosporiella-like synasexual morph in culture (Giraldo et al. 2014a).

Morphologically, A. grakistii resembles A. kalrae and A. curvata in having cylindrical arthroconidia and a trichosporiella-like synasexual morph. Arthrographis kalrae and A. curvata are able to grow at 37 °C (Sigler & Carmichael 1983, Giraldo et al. 2014a), while A. grakistii does not grow at this temperature.

Authors: A. Giraldo López and M. Hernández-Restrepo

Kramasamuha Subram. & Vittal, Canad. J. Bot. 51: 1128. 1973.

Conidiophores erect, flexuous, solitary to fasciculate, arising from a swollen basal cell, which appears lobed due to rhizoids; medium brown, smooth, multi-septate, simple or branched, giving rise to parallel stipes, becoming paler towards apex, terminating in an acute conidiogenous cell. Conidiogenous cells monoblastic, pale brown, ampulliform, straight to curved, tapering to a truncate apex, thin-walled, solitary or in clusters, integrated or discrete, terminal and intercalary. Conidia solitary, smooth, septate, obovoid to pyriform; second cell from base thick-walled, dark brown, somewhat swollen, basal and apical cell subhyaline, with short narrow separating cell at base as remnant from conidiogenous cell.

Type species: Kramasamuha sibika Subram. & Vittal

Kramasamuha sibika Subram. & Vittal, Canad. J. Bot. 51: 1129. 1973. Fig. 7.

Fig. 7.

Kramasamuha sibika (CBS 146133). A–C. Conidiophores overview on OA. D–F. Conidiophores, conidiogenous cells and conidia. G. Conidiogenous cells and conidia. H–J. Conidiogenous cells. K–N. Conidia. Scale bars: D–F = 25 μm, all others = 10 μm.

Typus: India, Tamil Nadu, Chingleput district, Vandalur, on dead leaves of Gymnosporia emarginata (Celastraceae), 23 Apr. 1971, B.P.R. Vittal, Herb. MUBL 2153 (not seen).

Occurring on leaf litter. Conidiophores erect, flexuous, solitary to fasciculate, arising from a swollen basal cell, 10–14 μm diam, which appears lobed due to rhizoids; medium brown, thin-walled, smooth, multi-septate, septa 17–30 μm apart, up to 550 μm tall, 3–5 μm diam, unbranched or branched, giving rise to parallel stipes, becoming paler towards apex, terminating in an acute conidiogenous cell. Conidiogenous cells monoblastic, pale brown, ampulliform, straight to curved, tapering to a truncate apex, thin-walled, solitary or in clusters of 2–4, integrated or discrete, terminal and intercalary, 4–8 × 3–4 μm. Conidia solitary, smooth, (1–)2(–3)-septate, blastoconidia, obovoid to pyriform, apex obtuse, (18–)25–27(–34) × (10–) 11(–12) μm; second cell from base thick-walled, dark brown, somewhat swollen, basal and apical cell subhyaline, with short narrow separating cell at base as remnant from conidiogenous cell, 1–1.5 × 1 μm.

Culture characteristics: Colonies on OA with scarce aerial mycelium, cottony to velvety, grey olivaceous, submerged mycelium vinaceous, margin effuse, irregular, reverse vinaceous. Conidiophores, conidiogenous cells, and conidia very similar to those observed in natural substrate. Conidiogenous cells 4.5–9 × 2.5–4 μm. Conidia 14–32.5 × 9–14 μm, (0–)2(–3)-septate.

Materials examined: Australia, New South Wales, Mallanganee, on leaves of Lophostemon confertus (Myrtaceae), 17 Apr. 2018, A.J. Carnegie, CBS 146338. Brazil, state of Minas Gerais, Viçosa, on Hypericum innodorum (Hypericaceae) leaves bearing necrotic spots caused by Seimatosporium hypericinum (Pinaceae), 4 Jul. 2017, A.A. Colmán (VIC 47176, COAD 2632). Malaysia, on needles of Pinus tecunumanii, Oct. 2018, M.J. Wingfield, CBS 146339. South Africa, KwaZulu-Natal, Richards Bay, on living leaves of Syzygium cordatum (Myrtaceae), Jun. 2016, M.J. Wingfield, CBS 146133 = CPC 36153.

Notes: Kramasamuha resembles the genus Garnaudia and some species of Endophragmiella in having conidia with a short narrow separating cell at the base as remnant from the conidiogenous cell. However, species in these genera can be distinguished by the arrangement and colour of the conidiogenous cells. In Kramasamuha they are pale brown and solitary or in clusters along the conidiophores (Subramanian & Vittal 1973). In Garnaudia, the conidiogenous cells are brown, and verticillate in terminal branches (Borowska 1977), while in Endophragmiella the conidiogenous cells are hyaline and mainly solitary and terminal (Hughes 1979, Seifert et al. 2011). Kramasamuha is a monotypic genus originally described from India on Gymnosporia emarginata (Subramanian & Vittal 1973). However, the specimens examined here were from different substrates and continents, i.e. Lophostemon confertus in Australia, Hypericum×innodorum in Brazil, Pinus tecunumanii in Malaysia, and Syzygium cordatum in South Africa and can thus not serve as an epitype. This species has also been recorded from leaves on Feijoa sellowiana in New Zealand, and on Psidium guajava in Western Samoa (Landcare database 2019).

This is the first time that DNA sequence data has become available for K. sibika and shows that Kramasamuha is related to Helminthosphaeriaceae (Sordariomycetes, Fig. 2). Asexual morphs in Helminthosphaeria have been recognised as diplococcium-like (Samuels et al. 1997, Réblová 1999) with tretic conidiogenous cells, differing from the monoblastic conidiogenous cells observed in Kramasamuha (Seifert et al. 2011).

Authors: M. Hernández-Restrepo, P.W. Crous, M.J. Wingfield, A.A. Colmán, P.S.C. Mansur and R.W. Barreto

Melnikomyces Crous & U. Braun, Persoonia 32: 263. 2014.

Mycelium consisting of brown, septate, branched, smooth, thick-walled hyphae. Conidiophores subcylindrical, brown, smooth, erect, straight or geniculate, reduced to conidiogenous cells, or long, flexuous, multiseptate. Conidiogenous cells polyblastic, subcylindrical to subclavate, terminal or intercalary, brown, smooth, developing a rachis with numerous denticle-like loci. Conidia solitary, brown, verruculose, fusoid-ellipsoidal, 1-septate, ends sub-obtuse, released by rhexolytic secession. Chlamydospores terminal, globose to subglobose, in short chains, simple or branched, brown, smooth (modified from Crous et al. 2014b).

Type species: Melnikomyces vietnamensis Crous & U. Braun

Melnikomycesthailandicus Giraldo López, sp. nov. MycoBank MB833678. Fig. 8.

Fig. 8.

Melnikomyces thailandicus sp. nov. (CBS 145767). A–C. Conidiophores and denticulated conidiogenous cells. D. Conidia. E. Chlamydospores. Scale bars = 10 μm.

Etymology: Name refers to Thailand where the fungus was collected.

Mycelium consisting of brown, septate, branched, smooth, thick-walled, 2–2.5 μm diam hyphae. Conidiophores macronematous, arising directly from vegetative hyphae, erect, straight or slightly bent, simple, multiseptate, cylindrical, 14–37 × 2–3 μm, brown, paler apex, thick and smooth-walled. Conidiogenous cells integrated, terminal, polyblastic, brown to pale brown, sympodial, with long open denticles; denticles cylindrical, pale brown, up to 1 μm long. Conidia fusoid, ends subobtuse, 1-septate, solitary, subhyaline, smooth-walled, (8–)9.5–12(–13) × (2–)2.5(–3) μm. Chlamydospores lateral, globose to subglobose, in short and simple chains, light brown, thick-and smooth-walled, 5.5–10 μm diam.

Typus: Thailand, Nakhon Nayok Province, Mueang Nakhon Nayok district, Wang Takrai waterfall, N14.330023° E101.307168°, 64 m above sea level, from soil, 22 Jul. 2008, P.W. Crous (holotype CBS H-24236, culture ex-type CBS 145767).

Notes: The monotypic genus Melnikomyces was introduced by Crous and Braun (Crous et al. 2014b), based on M. vietnamensis collected from dry leaves in Vietnam. This species was treated as incertae sedis in the Chaetothyriales, Eurotiomycetes (Crous et al. 2014b). However, the results of this study show that it resides in the Sympoventuriaceae (Venturiales, Dothideomycetes), together with other genera producing septate conidia from denticulate conidiogenous cells, such as Ochroconis and Verruconis (Machouart et al. 2014, Samerpitak et al. 2014).

Melnikomyces thailandicus is the second species described in the genus, which differs morphologically from M. vietnamensis in having shorter conidiophores (10–60 μm long vs.14–37 μm long) and longer and narrower smooth-walled conidia (8–13 × (2–3) μm vs. 7–11 × 2.5–3.5 μm). Originally, M. vietnamensis was described with two types of conidiophores (Crous et al. 2014b), but in M. thailandicus one of the conidiophore types more closely resembles chlamydospores, as they appear to stay attached to the hyphae.

Authors: A. Giraldo López and P.W. Crous

Thysanorea Arzanlou, W. Gams & Crous, Stud. Mycol. 58: 80. 2007. emend.

Conidiophores micro- or macronematous, erect, simple or apically branched, sometimes proliferating percurrently in the apex, brown, smooth. Conidiogenous cells terminal or intercalary, polyblastic, smooth, brown at the base, paler towards the apex, subcylindrical, clavate to doliiform, sympodial, with crowded conidiogenous loci inconspicuous to slightly prominent, refractive to somewhat obscure, slightly thickened. Conidia solitary, oblong, obovoid, cylindrical, broadly fusiform to subpyriform, pale brown, smooth, with a narrowly truncate base and darkened hilum; conidial secession schizolytic. Synasexual morph: Conidiophores erect, simple, brown, smooth. Conidiogenous cells terminal, discrete, phialidic, subglobose to lageniform, with a balloon- to funnel-shaped collarette, brown, often in clusters at the apex of the conidiophores. Conidia solitary, subglobose to obovate, unicellular, hyaline, guttulate, smooth.

Type species: Thysanorea papuana (Aptroot) Arzanlou et al.

Notes: Thysanorea was established for a genus similar to Periconiella based on the branching pattern of the conidiophores (Arzanlou et al. 2007). However, recent studies have shown that those branching patterns depend on culture conditions, where those on natural substrates or in young cultures are not as prominently branched as previously described (Kirschner 2016, Wang et al. 2019). The generic description is emended here to include species with a phialidic synasexual morphs.

Phylogenetically, Thysanorea is closely related to Minimelanolocus in the Chaetothyriales (Fig. 3). However, the phylogenetic position of M. navicularis, the type species, remains unknown since DNA sequence data are not available for it, and the supposed phylogenetic position has been based on other species (Liu et al. 2015, Wang et al. 2019).

Morphologically, M. navicularis is characterised by terminal conidiogenous cells that produce navicular conidia with sub-hyaline polar cells and darker central cells (Castañeda-Ruiz 1987). They consequently differ from those species placed in Minimelanolocus based on DNA sequence data and in which the conidiogenous cells are terminal and intercalary, and the conidia are oblong, obovoid, cylindrical, broadly fusiform, and uniformly pale brown (Liu et al. 2015, Wang et al. 2019). In this regard, they would fit better with the generic concept of Thysanorea (Arzanlou et al. 2007, Wang et al. 2019). Based on these morphological differences and phylogenetic relationships, we propose new combinations for those species that have been shown as related to Thysanorea. The phylogenetic placement of other species for which DNA sequence data are not available, including M. navicularis must still be determined.

Thysanorea asiatica (H.Y. Su, et al.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833919.

Basionym: Minimelanolocus asiaticus H.Y. Su, et al., Fungal Biol. 119: 1054. 2015.

Thysanorea cantrelliae Hern.-Restr., R. van Doorn & Crous, sp. nov. MycoBank MB833914. Fig. 9.

Fig. 9.

Thysanorea cantrelliae sp. nov. (CBS 145909). A. Conidiophore and conidium. B, C. Conidiophores. D–G. Conidiogenous cells. H, I. Conidia. Scale bars = 10 μm.

Etymology: Named in honour of Sharon Cantrell, who was the organizer of the IMC 11 (2018) in Puerto Rico. This fungus was collected on a field trip held during the IMC 11.

Mycelium composed of hyaline to pale brown, septate, smooth, 1–2 μm wide hyphae. Conidiophores semi-micronematous, sometimes reduced to conidiogenous cells, simple, erect, straight or flexuous, cylindrical, pale brown, smooth, 8–31 × 2–3 μm. Conidiogenous cells holoblastic, polyblastic, mainly terminal, integrated, sympodial, pale brown, 7–22 × 2–3 μm. Conidia solitary, fusiform to acicular, straight or curved, (1–)3(–4)-septate, subhyaline to pale brown, smooth, 10–34 × 1.5–3 μm, apex acute, base truncate.

Culture characteristics: Colonies at 25 °C after 14 d: on OA reaching 22–25 mm, aerial mycelium moderate, cottony to floccose, olivaceous black, margin effuse entire; reverse black. On MEA and PDA reaching 22–30 mm, aerial mycelium abundant, cottony to floccose, olivaceous grey, black close to the agar, margin effuse, entire; reverse black.

Typus: USA, Puerto Rico, from unidentified twig, 20 Jul. 2018, M. Hernández-Restrepo (holotype CBS H-24100, culture ex-type CBS 145909).

Notes: Thysanorea cantrelliae clustered in a separate clade together with T. seifertii (Fig. 3). It can be distinguished from T. seifertii and other species in the genus by its acicular conidia (Fig. 9).

Thysanorea curvata (H.Y. Su et al.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833921.

Basionym: Minimelanolocus curvatus H.Y. Su et al., Fungal Biol. 119: 1055. 2015.

Thysanorea lotorum (Morgan-Jones) Hern.-Restr. & Crous, comb. nov. MycoBank MB833922.

Basionym: Pseudospiropes lotorum Morgan-Jones, Mycotaxon 5: 481. 1977 [as ‘lotorus’]

Synonym: Nigrolentilocus lotorum (Morgan-Jones) R.F. Castañeda & Heredia, Cryptog. Mycol. 22: 15. 2001.

Minimelanolocus submersus Z.L. Luo et al., Fungal Diversity 80: 143. 2016.

Thysanorea melanica (H.Y. Su, et al.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833923.

Basionym: Minimelanolocus melanicus H.Y. Su et al., Fungal Biol. 119: 1056. 2015.

Thysanorea nonramosa (X.D. Yu et al.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833924.

Basionym: Minimelanolocus nonramosus X.D. Yu et al., Mycol. Progr. 18: 514. 2019.

Thysanorea obscura (Matsush.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833925.

Basionym: Pseudospiropes obscurus Matsush., Matsushima Mycol. Mem. 3: 14. 1983.

Synonym: Minimelanolocus obscurus (Matsush.) R.F. Castañeda & Heredia, Cryptog. Mycol. 22:10. 2001.

Thysanorea rousseliana (Mont.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833926.

Basionym: Helminthosporium rousselianum Mont., Ann. Sci. Nat., Sér. 3, Bot., 12: 300. 1849.

Synonyms: Pleurophragmium rousselianum (Mont.) S. Hughes, Canad. J. Bot. 36: 798. 1958.

Spiropes rousselianus (Mont.) de Hoog & Arx, Kavaka 1: 59. 1973.

Pseudospiropes rousselianus (Mont.) M.B. Ellis, More Dematiaceous Hyphomycetes: 221. 1976.

Minimelanolocus rousselianus (Mont.) R.F. Castañeda & Heredia, Cryptog. Mycol. 22: 10. 2001.

Thysanorea seifertii Hern.-Restr., R. van Doorn & Crous, sp. nov. MycoBank MB833920. Fig. 10.

Fig. 10.

Thysanorea seifertii sp. nov. (CBS 145910). A. Conidiophores and conidia. B, C. Conidiogenous cells and conidia. D. Conidia. E–J. Synasexual morph. E, F. Conidiophores. G–I. Conidiogenous cells and conidia. H. Conidia. Scale bars: G–J = 5 μm, all others = 10 μm.

Etymology: Named in honour of Prof. dr Keith A. Seifert, who was President of the International Mycological Association during the IMC 11 (2018) in Puerto Rico. This fungus was collected on a field trip during the IMC 11.

Mycelium composed of hyaline to pale brown, septate, smooth, 1–2 μm wide hyphae. Conidiophores mononematous, simple, erect, straight or flexuous, cylindrical, brown, paler towards the apex, smooth, 30–133 × 2–3 μm. Conidiogenous cells holoblastic, polyblastic, terminal or intercalary, integrated, sympodial, pale brown to brown, 7.5–46.5 × 2–3 μm. Conidia solitary, subcylindrical to clavate or oblong, straight or slightly curved, (0–)1–3(–4)-septate, pale brown, smooth, 7–15 × 1.5–3 μm, apex rounded, base darkened and truncated. Synasexual morph: Conidiophores micro- or macronematous, erect, straight or flexuous, cylindrical, brown, smooth, 10–51 × 2–4 μm. Conidiogenous cells enteroblastic, phialidic, arranged around the apex of the conidiophore, brown, subglobose to ampulliform, 2–4(–6) × 2–3 μm, with a balloon- to funnel-shaped collarette, 1–3 × 1–3 μm. Conidia solitary, subglobose to obovate, unicellular, hyaline, guttulate, smooth, 1–2 × 1–1.5 μm, base truncated.

Culture characteristics: Colonies at 25 °C after 14 d: on OA reaching 22–25 mm, aerial mycelium moderate, cottony to floccose, grey olivaceous to olivaceous black, margin effuse, entire; reverse black. On MEA and PDA reaching 18–20 mm, aerial mycelium moderate to abundant, cottony to floccose, olivaceous grey, black close to the agar, margin effuse, entire; reverse black.

Typus: USA, Puerto Rico, from unidentified twig, 20 Jul. 2018, M. Hernández-Restrepo (holotype CBS H-24101, culture ex-type CBS 145910).

Notes: Some of the conidia in T. seifertii resemble those of T. obscura in being 3-septate. However, conidia in T. seifertii are smaller than those of T. obscura (7–15 × 1.5–3 μm vs. 20–31 × 5–8 μm, Castañeda-Ruiz et al. 2001). Thysanorea seifertii is the only species in the genus known to produce a phialidic synasexual morph.

Thysanorea thailandensis (W. Dong et al.) Hern.-Restr. & Crous, comb. nov. MycoBank MB833927.

Basionym: Minimelanolocus thailandensis W. Dong et al., Mycol. Progr. 17: 622. 2018.

Thysanorea yunnanensis Hern.-Restr. & Crous, nom. nov. MycoBank MB833928.

Replaced synonym: Minimelanolocus aquaticus H.Y. Su et al., Fungal Biol. 119: 1049. 2015 [non Thysanorea aquatica W. Dong, H. Zhang & K.D. Hyde, 2018].

Etymology: The name refers to the Chinese Province of Yunnan where the fungus was collected.

Authors: M. Hernández-Restrepo, R. van Doorn and P.W. Crous

Verruconis Samerp. et al., Fungal Diversity 65: 117. 2014.

Mycelium consisting of septate, pale brown, smooth and thick-walled hyphae. Conidiophores differentiated, erect, straight or slightly bent, unbranched, pale brown. Conidiogenous cells mostly polyblastic, subcylindrical to narrowly mucronate, producing conidia sympodially on long open denticles; denticles cylindrical, pale brown, scattered at the apical third of the conidiogenous cell. Conidia two-celled, ellipsoidal, cylindrical or clavate, brown, verrucose or smooth-walled, released by rhexolytic secession. Sexual morph unknown.

Type species: Verruconis gallopava (W.B. Cooke) Samerp. & de Hoog

Verruconis thailandica Giraldo López & Crous, sp. nov. MycoBank MB833679. Fig. 11.

Fig. 11.

Verruconis thailandica sp. nov. (CBS 145768). A, B. Colonies on PDA and OA, respectively, at 25 °C after 14 d. C–E. Denticulated conidiogenous cells. F. Conidia with gelatinous brown sheath (arrow). Scale bars = 10 μm.

Etymology: The name refers to Thailand where the fungus was collected.

Mycelium consisting of septate, pale brown, smooth, thick-walled, 2–2.5 μm diam hyphae. Conidiophores differentiated, arising directly from vegetative hyphae, erect, straight or slightly bent, simple, 0–1-septate, subcylindrical, (3.6–)4.1–7.1(–9) × (1.3–)2(–2.3) μm, pale brown, thick- and smooth-walled, producing conidia sympodially on long open denticles; denticles cylindrical, pale brown, up to 1 μm long. Conidia abundant on OA and PCA, scarce on PDA, two-celled, broadly ellipsoidal with a protuberant hilum, constricted at the septum, (5–)5.8(–7) × (2.2–)2.6(–3.1) μm, brown, verrucose, thick-walled, sometimes with a wing-like gelatinous brown sheath, released by rhexolytic secession. Sexual morph not observed.

Cultural characteristics: Colonies at 25 °C after 14 d: on OA and PCA, flat, woolly at centre, glabrous at periphery, top and reverse sepia. On PDA raised, felty, top and reverse olivaceous with ochreous diffusible pigment.

Typus: Thailand, Nakhon Nayok Province, Mueang Nakhon Nayok district, Wang Takrai waterfall, N14.330023° E101.307168°, 64 m above sea level, from soil, 22 Jul. 2008, P.W. Crous (holotype CBS H-24237, culture ex-type CBS 145768).

Notes: The genus Verruconis (Sympoventuriaceae, Venturiales, Dothideomycetes) was established to accommodate thermophilic species segregated from Ochroconis (O. gallopava and O. calidifluminalis) and Scolecobasidium (S. verruculosum), which produce septate conidia from sympodially proliferating conidiophores, released by rhexolytic secession (Samerpitak et al. 2014). These species have been isolated from hot spring water, warm effluents or as soil saprobes (Yarita et al. 2007, Samerpitak et al. 2014, Giraldo et al. 2014b). However, the type species, V. gallopava has been reported as an opportunistic pathogen of humans and causing infections in other warm-blooded animals, mainly birds (Revankar & Sutton 2010, de Hoog et al. 2011).

Verruconis thailandica is phylogenetically related to V. verruculosa (Fig. 1), but the two species can be distinguished by the length of their conidiophores (up to 9 μm long in V. thailandica vs. up to 45 μm long in V. verruculosa) and conidia (up to 7 μm long in V. thailandica vs. up to 9 μm long in V. verruculosa; Samerpitak et al. 2014). Recently, three new species were added to the genus, namely V. panacis from Panax notoginseng (Zhang et al. 2018), V. hainanensis and V. pseudotricladiata from submerged decaying leaves (Qiao et al. 2019). These species clustered in a separate clade phylogenetically distant from V. thailandica. They can be distinguished by the presence of four-celled conidia in V. panacis and V. hainanensis, and branched Y-shaped conidia in V. pseudotricladiata (Zhang et al. 2018, Qiao et al. 2019).

Authors: A. Giraldo López and P.W. Crous