Funiliomycetaceae fam. nov. (Amphisphaeriales, Ascomycota) accommodating Funiliomyces, including F. jiangxiensis sp. nov. from Tetradium ruticarpum and ten new combinations

Abstract

Figures

Maximum likelihood phylogenetic tree of Pezizomycotina based on LSU sequences, with Shimodaira-Hasegawa-like approximate likelihood ratio test (SH-aLRT) (left), ultrafast bootstrap (UFB) (middle), and Bayesian posterior probabilities (BPP) values (right) near the corresponding node. Only one of SH-aLRT > 80 or UFB > 95 for ML and BYPP > 0.90 for BI is indicated along the branches (SH-aLRT/UFB/BPP). The “Dactylaria” species are indicated in red, the newly generated sequences are indicated in green, and species for reclassification from Dactylaria to Funiliomyces are in blue. Ex-type strains are marked with T after the strain number. The ML phylogram is available in TreeBASE (study accession S32460; http://purl.org/phylo/treebase/phylows/study/TB2:S32460).

Maximum likelihood phylogenetic tree of Amphisphaeriales based on combined LSU, ITS, and RPB2 sequences, with Shimodaira-Hasegawa-like approximate likelihood ratio test (SH-aLRT) (left), ultrafast bootstrap (UFB) (middle), and Bayesian posterior probabilities (BPP) values (right) near the corresponding node. Only one of SH-aLRT > 80 or UFB > 95 for ML and BYPP > 0.90 for BI is indicated along the branches (SH-aLRT/UFB/BPP). Ex-type strains are marked with T after the strain number. The newly generated sequences are indicated in red and species for reclassification are in blue. The ML phylogram is available in TreeBASE (study accession S32462; http://purl.org/phylo/treebase/phylows/study/TB2:S32462).

Funiliomyces jiangxiensis sp. nov. (HFJAU10125, holotype) a, b Colonies on the front and back of PDA medium (for 5 days); c Hyphae and sporulation structures; d, e Conidiophores bearing a conidium initial on one of its denticles; f, g Conidiophores; h–l Conidia. Scale bars: 10 μm (c, d); 5 μm (e); 10 μm (f, g); 20 μm (h); 10 μm (i–l).

Tables

Table 1.. Taxa, isolate information, and GenBank accession numbers for sequences used in the phylogenetic analyses of Pezizomycotina.

Taxon	Strain	GenBank accession numbers	References
		LSU
Acarosporina microspora	CBS 338.39	AY584643	Lutzoni et al. (2004)
Amphisphaeria umbrina	HKUCC 994	AF452029	Jeewon et al. (2003)
Anungitiomyces stellenboschiensis	CPC 34726T	NG_067874	Crous et al. (2019a)
Appendicospora hongkongensis	HKAS 107015	MW240581	Samarakoon et al. (2022)
Arthonia dispersa	UPSC 2583	AY571381	Lumbsch et al. (2005)
Arthrobotrys macroides	CBS 120.54	MH868799	Vu et al. (2019)
Arthrobotrys mangrovispora	MGDW17	EU573355	Swe et al. (2008)
Ascosphaera apis	CBS 402.96	FJ358275	Gueidan et al. (2008)
Aspergillus fumigatus	INFU/Jc/KF/6	FM179606	Kusari et al. (2009)
Barrmaelia serenoae	CPC:37572T	MT223876	Crous et al. (2020)
Caliciopsis pinea	CBS 139.64	DQ678097	Schoch et al. (2006b)
Camarops ustulinoides	AFTOL-ID 72	DQ470941	Spatafora et al. (2006)
Candelaria concolor	AFTOL-ID 1706	DQ986791	Miadlikowska et al. (2006)
Candelariella reflexa	AFTOL-ID 1271	DQ912331	Miadlikowska et al. (2006)
Candelariella terrigena	AFTOL-ID 227	DQ986745	Miadlikowska et al. (2006)
Candida albicans	ATCC 18804T	NG_054826	Suh et al. (2013)
Capnodium coffeae	CBS 147.52	DQ247800	Schoch et al. (2006a)
Chaenotheca furfuracea	Wedin 6366 (UPS)T	JX000087	Prieto et al. (2013)
Cirrosporium novae-zelandiae	CBS 125236	HQ878612	Réblová et al. (2012)
Cladonia caroliniana	AFTOL-ID 3	AY584640	Lutzoni et al. (2004)
Cladosporium herbarum	CBS 121621	MH874676	Vu et al. (2019)
Coccomyces strobi	AFTOL-ID 1250	DQ470975	Spatafora et al. (2006)
Collemopsidium angermannicum	s1473	KU556871	Pérez-Ortega et al. (2016)
Collemopsidium pelvetiae	RO25	KU556868	Pérez-Ortega et al. (2016)
Conlarium baiseense	TD17	MK164655	Xie et al. (2019)
Cordyceps militaris	CBS 110.70	MH878247	Vu et al. (2019)
Corynelia africana	PREM 57242T	NG_058910	Wood et al. (2016)
Dactylaria acicularis	CBS 511.72	MH872253	Vu et al. (2019)
Dactylaria aquatica	P003	EU107312	Bhilabutra et al. (2007, submitted directly)
Dactylaria belliana	P020	EU107295	Bhilabutra et al. (2007, submitted directly)
Dactylaria dimorpha	P067	EU107305	Bhilabutra et al. (2007, submitted directly)
Dactylaria dimorphospora	CBS 256.70	MH871358	Vu et al. (2019)
Dactylaria flammulicornuta	P005	EU107313	Bhilabutra et al. (2007, submitted directly)
Dactylaria humicola	P006	EU107304	Bhilabutra et al. (2007, submitted directly)
Dactylaria hyalotunicata	P010	EU107298	Bhilabutra et al. (2007, submitted directly)
Dactylaria longispora	P001	EU107299	Bhilabutra et al. (2007, submitted directly)
Dactylaria lunata	CBS 689.93	MH874102	Vu et al. (2019)
Dactylaria parvispora	P024	EU107296	Bhilabutra et al. (2007, submitted directly)
Dactylaria purpurella	P048	EU107301	Bhilabutra et al. (2007, submitted directly)
Dactylaria sahelensis	CBS 247.93	MH874055	Vu et al. (2019)
Dactylaria simonensis	CBS 367.90	MH873904	Vu et al. (2019)
Dactylaria sporexamorpha	CBS 690.93	MH874103	Vu et al. (2019)
Dactylaria uliginicola	P007	EU107311	Bhilabutra et al. (2007, submitted directly)
Dactylellina appendiculata	CBS 206.64T	NG_059071	Li et al. (2006)
Dactylellina leptospora	CBS 560.92	AY261162	Crous et al. (2022)
Dactylospora haliotrepha	AFTOL-ID 758	FJ176855	Schoch et al. (2009a)
Dactylospora mangrovei	AFTOL-ID 2108	FJ176890	Schoch et al. (2009a)
Delphinella strobiligena	AFTOL-ID 1257	DQ470977	Spatafora et al. (2006)
Dendrographa minor	R. Ornduff 10070	AF279382	Bhattacharya et al. (2000)
Diatrype disciformis	AFTOL-ID 927	DQ470964	Spatafora et al. (2006)
Dothiora cannabinae	CBS 737.71	DQ470984	Spatafora et al. (2006)
Elaphomyces granulatus	AFTOL-ID 436T	KT232217	Voglmayr et al. (2019)
Eremascus albus	CBS 975.69	MH871279	Voglmayr et al. (2019)
Exophiala dermatitidis	AFTOL-ID 668T	DQ823100	Voglmayr et al. (2019)
Funiliomyces acaciae	CPC 29771T	KY173493	Crous et al. (2016)
Funiliomyces biseptatus	P062	EU107288	Bhilabutra et al. (2007, submitted directly)
Funiliomyces calliandrae	CPC 48004T	PV664963	Crous et al. (2025)
Funiliomyces fragilis	P057	EU107290	Bhilabutra et al. (2007, submitted directly)
Funiliomyces hwasunensis	CMML 20-35	PQ741487	Liu et al. (2025)
Funiliomyces jiangxiensis	CCTCC AF2023034T	OQ869216	This study
Funiliomyces mavisleverae	BRIP 76362aT	PQ431199	Tan and Shivas (2024)
Funiliomyces monticola	P060	EU107289	Bhilabutra et al. (2007, submitted directly)
Funiliomyces retrophylli	CBS:148271	ON811548	Crous et al. (2022)
Funiliomyces sparsus	P055	EU107291	Bhilabutra et al. (2007, submitted directly)
Funiliomyces zapatensis	P056	EU107287	Bhilabutra et al. (2007, submitted directly)
Fusarium graminearum	CBS 132906	MH877513	Vu et al. (2019)
Gelasinospora tetrasperma	CBS 178.33	DQ470980	Spatafora et al. (2006)
Geoglossum nigritum	AFTOL-ID 56T	AY544650	Schoch et al. (2009b)
Halosphaeria appendiculata	CBS 197.60	MH869504	Vu et al. (2019)
Herpomyces chaetophilus	D. Haelew. 1097b	MG438352	Haelewaters et al. (2019b)
Hypocrea lutea	ATCC 208838	AF543791	Currie et al. (2003)
Hyponectria buxi	UME 31430	AY083834	Aptroot et al. (2004)
Laboulbenia bruchii	D. Haelew 1346b	MN394843	Haelewaters et al. (2019a)
Lecanora contractula	AFTOL-ID 877	DQ986746	Miadlikowska et al. (2006)
Lecanora paramerae	Lumbsch s.n. (F)	EF105422	Crespo et al. (2007)
Lempholemma polyanthes	Zoladeski and Lutzoni 11294-L1(2/2) (CANL)	AF356691	Lutzoni et al. (2001)
Lepraria lobificans	AFTOL-ID 325	DQ986768	Miadlikowska et al. (2006)
Lichina confinis	I1024	MT877184	Garrido-Benavent (2020, submitted directly)
Lichina pygmaea	I1028	MT877182	Garrido-Benavent (2020, submitted directly)
Marcelleina persoonii	AFTOL-ID 164	DQ470943	Spatafora et al. (2006)
Microascus longirostris	CBS 196.61	NG_058479	Daros-Pawlyta and Pawlyta (2023)
Mycocalicium subtile	Wedin 6889 (UPS)T	AY853379	Wedin et al. (2005)
Mycosphaerella etlingerae	CBS 129062	NG_069080	Crous et al. (2011b)
Myrmecridium schulzeri	CBS 188.96T	EU041829	Arzanlo et al. (2007)
Nectria cinnabarina	CBS_114055	KU382228	Zare and Gams (2016)
Neomyrmecridium asiaticum	CBS:145080	NG_066291	Crous et al. (2018a)
Nothodactylaria nephrolepidis	CPC 37028T	NG_068668	Crous et al. (2019b)
Orbilia albidorosea	CBS 140818T	NG_073646	Vu et al. (2019)
Orbilia vinosa	AFTOL-ID 905T	DQ470952	Spatafora et al. (2006)
Peltula umbilicata	AFTOL-ID 891	DQ832334	James et al. (2006)
Petriella setifera	AFTOL-ID 956	DQ470969	Spatafora et al. (2006)
Peziza varia	JC19031901	MT247057	Van Vooren (2020)
Peziza vesiculosa	AFTOL-ID 507	DQ470948	Spatafora et al. (2006)
Plicaria leiocarpa	CBS 144.92	DQ842029	Spatafora et al. (2006)
Pseudodactylaria xanthorrhoeae	CBS:143414	NG_058521	Crous et al. (2017)
Pseudotruncatella arezzoensis	MFLUCC 14-0988T	NG_070426	Perera et al. (2018)
Pyrenula reebiae	DUKE:0047599T	NG_068722	Reeb et al. (2004)
Ramichloridium anceps	AFTOL-ID 659T	DQ823102	James et al. (2006)
Roccella fuciformis	AFTOL-ID 126T	AY584654	Lutzoni et al. (2004)
Roccellographa cretacea	AFTOL-ID 93	DQ883696	Spatafora et al. (2006)
Sabahriopsis eucalypti	CPC:24957	NG_058167	Crous et al. (2015b)
Saccharomyces cerevisiae	NRRL Y-12632T	NG_042623	Kurtzman et al. (2013)
Sarea difformis	JR6451T	MN938401	Beimforde et al. (2020)
Sarea resinae	JR6450	MN938402	Beimforde et al. (2020)
Schismatomma decolorans	Ertz 5003 (BR)	AY548815	Lutzoni et al. (2004)
Sclerophora farinacea	Wedin 6414 (UPS)	JX000095	Prieto et al. (2013)
Sordaria fimicola	SMH4106	AY780079	Miller and Huhndorf (2005)
Spathularia velutipes	AFTOL-ID 1291	FJ997861	Voglmayr et al. (2019)
Sphinctrina turbinata	AFTOL-ID 1721	EF413632	Geiser et al. (2006)
Teloschistes exilis	AFTOL-ID 87	AY584647	Lutzoni et al. (2004)
Toxicocladosporium chlamydosporum	CBS 124157T	NG_069916	Vu et al. (2019)
Trapelia placodioides	KS163	KU844623	Schneider et al. (2016)
Trichocoma paradoxa	CBS 788.83	FJ358290	Gueidan et al. (2008)
Trichoglossum hirsutum	AFTOL-ID 64	AY544653	Voglmayr et al. (2019)
Trinosporium guianense	CBS 132537T	NG_042682	Crous et al. (2012)
Tryblidiopsis pinastri	AFTOL-ID 1319	DQ470983	Spatafora et al. (2006)
Umbilicaria arctica	AFTOL-ID 1266	DQ986772	Miadlikowska et al. (2006)
Umbilicaria hyperborea	OSC:McCune 35477	MH290350	McCune (2018)
Volutella ciliata	CBS 127312	MH875955	Vu et al. (2019)
Xylaria acuta	AFTOL-ID 63	AY544676	Hongsanan and Hyde (2017)
Xylaria hypoxylon	AFTOL-ID 51	AY544648	Voglmayr et al. (2019)
Xylobotryum andinum	XA1	MH468791	Voglmayr et al. (2019)
Xylobotryum portentosum	XP	MH468792	Voglmayr et al. (2019)
Xylona heveae	CBS 132557T	NG_066160	Gazis et al. (2012)

Table 2.. Taxa, isolate information, and GenBank accession numbers for sequences used in the phylogenetic analyses of Amphisphaeriales.

Taxon	Strain	GenBank accession numbers			References
		LSU	ITS	RPB2
Achaetomium macrosporum	CBS 532.94	KX976699	KX976574	KX976797	Wang et al. (2016)
Amphisphaeria fuckelii	CBS:140409T	KT949902	KT949902	MH554918	Jaklitsch et al. (2016)
Amphisphaeria thailandica	MFLU 18-0794T	MH971235	MH971225	MK033640	Samarakoon et al. (2019)
Anungitiomyces stellenboschiensis	CPC:34726T	MK876415	MK876376	NA	Crous et al. (2019b)
Appendicospora hongkongensis	HKAS 107015	MW240581	MW240651	MW658638	Samarakoon et al. (2022)
Arthrinium hysterinum	ICMP 6889	MK014841	MK014874	DQ368649	Pintos et al. (2019)
Arthrinium pseudoparenchymaticum	SICAUCC 18-0008	MK346321	MK346319	MK359207	Yang et al. (2019)
Beltrania pseudorhombica	CBS:138003T	KJ869215	MH554124	MH555032	Crous et al. (2014a)
Beltraniopsis neolitseae	CBS:137974T	KJ869183	KJ869126	NA	Crous et al. (2014a)
Brachiampulla verticillata	ICMP 15993	MW144403	MW144419	NA	Réblová et al. (2021)
Castanediella acaciae	CBS 139896T	MH878661	KR476728	NA	Vu et al. (2019)
Castanediella cagnizarii	MUCL 41095	KC775707	KC775732	NA	Becerra-Hernández et al. (2016)
Castanediella ramosa	MUCL 39857	KC775711	KC775736	NA	Becerra-Hernández et al. (2016)
Chaetomium elatum	CBS 374.66	MH870466	KC109758	KF001820	Vu et al. (2019)
Clypeophysalospora latitans	CBS 141463T	KX820261	KX820250	NA	Giraldo et al. (2017)
Cylindrium elongatum	CBS 115974	KM231733	KM231853	KM232429	Lombard et al. (2015)
Cylindrium grande	CBS:145578	MK876426	MK876385	MK876482	Crous et al. (2019a)
Funiliomyces acaciae	CPC 29771	KY173493	KY173400	NA	Crous et al. (2016)
Funiliomyces biseptatus	P062	EU107288	NA	NA	Bhilabutra et al. (2007, submitted directly)
Funiliomyces biseptatus	CBS 100373T	NG_067443	NR_159862	NA	Aptroot (2004)
Funiliomyces calliandrae	CPC 48004	PV664963	PV664937	PV664022	Crous et al. (2025)
Funiliomyces fragilis	P057	EU107290	NA	NA	Bhilabutra et al. (2007, submitted directly)
Funiliomyces hwasunensis	CMML 20-35	PQ741487	PQ741486	NA	Liu et al. (2025)
Funiliomyces hwasunensis	CMML 20-88	PQ741488	NA	NA	Liu et al. (2025)
Funiliomyces jiangxiensis	CCTCC AF2023034T	OQ869216	OQ869213	OR046688	This study
Funiliomyces jiangxiensis	CCTCC AF2023033	OQ869214	OQ869215	OR046687	This study
Funiliomyces mavisleverae	BRIP 76362a	PQ431199	PQ431206	NA	Tan and Shivas (2024)
Funiliomyces monticola	P060	EU107289	NA	NA	Bhilabutra et al. (2007, submitted directly)
Funiliomyces retrophylli	CBS:148271	ON811548	ON811489	NA	Crous et al. (2022)
Funiliomyces sparsus	P055	EU107291	NA	NA	Bhilabutra et al. (2007, submitted directly)
Funiliomyces zapatensis	P056	EU107287	NA	NA	Bhilabutra et al. (2007, submitted directly)
Hyponectria buxi	UME 31430	AY083834	NA	NA	Smith et al. (2002, submitted directly)
Iodosphaeria honghensis	MFLU 19-0719T	MK722172	MK737501	MK791287	Marasinghe et al. (2019)
Iodosphaeria tongrenensis	MFLU:15-0393	KR095283	KR095282	NA	Li et al. (2015)
Leiosphaerella praeclara	CBS:125586	JF440976	JF440976	NA	Jaklitsch and Voglmayr (2012)
Melogramma campylosporum	MFLU 17-0348	MW240575	MW240645	MW658632	Samarakoon et al. (2022)
Melogramma campylosporum	MFLU 18-0778	MW240576	MW240646	MW658633	Samarakoon et al. (2022)
Neoamphisphaeria hyalinospora	MFLU 19-2131T	MW240579	MW240649	MW658636	Samarakoon et al. (2022)
Neoamphisphaeria shangrilaensis	HKAS 130274	PP584800	PP584703	NA	Dissanayake et al. (2024)
Neophysalospora eucalypti	CBS:138864T	KP004490	KP004462	NA	Crous et al. (2014b)
Nothodactylaria comitabilis	CPC 45173	PQ498974	PQ498925	NA	Crous et al. (2024)
Nothodactylaria fusiformis	KUNCC:23-13961T	PQ671162	PQ671242	PQ662509	Zhang et al. (2025)
Nothodactylaria guizhouensis	KUNCC:23-14080T	PQ671163	PQ671243	PQ662510	Zhang et al. (2025)
Nothodactylaria nephrolepidis	CPC:37028	MN567639	MN562132	MN556809	Crous et al. (2019b)
Nothodactylaria nephrolepidis	CBS:146078T	NG_068668	NR_166331	NA	Crous et al. (2019b)
Nothodactylaria polyblastis	KUNCC:23-13922T	PQ671164	PQ671244	PQ662511	Zhang et al. (2025)
Nothodactylaria woodwardiae	KUNCC:23-13927T	PQ671165	PQ671245	PQ662512	Zhang et al. (2025)
Nothodactylaria woodwardiae	KUNCC:23-13886	PQ671166	PQ671246	PQ662513	Zhang et al. (2025)
Nothodactylaria woodwardiae	KUNCC:23-14045	PQ671167	PQ671247	PQ662514	Zhang et al. (2025)
Nothodactylaria woodwardiae	KUNCC:23-13954	PQ671168	PQ671248	NA	Zhang et al. (2025)
Nothodactylaria woodwardiae	KUNCC23-14006	PQ671169	PQ671249	NA	Zhang et al. (2025)
Oxydothis metroxyli	MFLUCC15-0283	KY206764	KY206775	NA	Konta et al. (2016)
Oxydothis metroxylonicola	MFLUCC 15-0281T	KY206763	KY206774	KY206781	Konta et al. (2016)
Oxydothis palmicola	MFLUCC 15-0806T	KY206765	KY206776	KY206782	Konta et al. (2016)
Phlogicylindrium eucalypti	CBS 120080T	DQ923534	DQ923534	MH554893	Summerell et al. (2006)
Phlogicylindrium uniforme	CBS 131312T	JQ044445	JQ044426	NA	Crous et al. (2011a)
Polyscytalum eucalyptorum	CPC 17207T	KJ869176	KJ869118	NA	Crous et al. (2014a)
Pseudapiospora corni	WU:36852T	KT949907	KT949907	NA	Jaklitsch et al. (2016)
Pseudomassaria chondrospora	CBS:125600	JF440981	JF440981	NA	Jaklitsch and Voglmayr (2012)
Pseudosporidesmium knawiae	CBS:123529T	MH874823	MH863299	NA	Vu et al. (2019)
Pseudosporidesmium lambertiae	CBS:143169T	MG386087	MG386034	NA	Crous et al. (2017)
Pseudotruncatella arezzoensis	MFLUCC:14-0988T	MG192317	MG192320	NA	Perera et al. (2018)
Pseudotruncatella bolusanthi	CBS:145532T	MK876448	MK876407	NA	Crous et al. (2019a)
Robillarda sessilis	CBS 114312T	KR873284	KR873256	NA	Crous et al. (2015a)
Seiridium marginatum	CBS:140403T	KT949914	KT949914	MK523301	Jaklitsch et al. (2016)
Sordaria fimicola	CBS 723.96	NA	MH862606	NA	Vu et al. (2019)
Strelitziomyces knysnanus	CPC:37067T	MN567642	MN562135	MN556810	Crous et al. (2019b)
Subsessila turbinata	MFLUCC:15-0831T	KX762289	KX762288	NA	Lin et al. (2017)
Vialaea insculpta	DAOM:240257	JX139726	JX139726	NA	Hambleton et al. (2010, submitted directly)
Vialaea minutella	BRIP:56959	KC181924	KC181926	NA	McTaggart et al. (2013)
Xyladictyochaeta lusitanica	CBS:142290T	KY853543	KY853479	NA	Hernández-Restrepo et al. (2017)

Table 3.. Estimates of evolutionary divergence in ITS rDNA, LSU rDNA, and RPB2 sequences.

ITS rDNA
	Beltraniaceae	Castanediellaceae	Clypeophysalosporaceae	Amphisphaeriaceae	Sporocadaceae	Pseudomassariaceae	Phlogicylindriaceae	Xyladictyochaetaceae	Cylindriaceae	Vialaeaceae	Nothodactylariaceae	Funiliomycetaceae	Pseudotruncatellaceae	Melogrammataceae	Appendicosporaceae	Apiosporaceae	Oxydothidaceae	Anungitiomycetaceae	Pseudosporidesmiaceae
Castanediellaceae	0.110
Clypeophysalosporaceae	0.181	0.168
Amphisphaeriaceae	0.145	0.157	0.217
Sporocadaceae	0.125	0.119	0.144	0.167
Pseudomassariaceae	0.187	0.185	0.166	0.209	0.157
Phlogicylindriaceae	0.150	0.145	0.185	0.178	0.145	0.186
Xyladictyochaetaceae	0.137	0.122	0.189	0.162	0.128	0.173	0.136
Cylindriaceae	0.140	0.132	0.130	0.150	0.152	0.150	0.136	0.121
Vialaeaceae	0.240	0.239	0.228	0.255	0.225	0.213	0.238	0.259	0.242
Nothodactylariaceae	0.250	0.233	0.237	0.270	0.223	0.183	0.223	0.244	0.195	0.238
Funiliomycetaceae	0.230	0.229	0.230	0.264	0.217	0.182	0.223	0.229	0.209	0.231	0.156
Pseudotruncatellaceae	0.178	0.154	0.147	0.179	0.130	0.158	0.181	0.159	0.167	0.212	0.206	0.219
Melogrammataceae	0.227	0.214	0.209	0.255	0.159	0.179	0.204	0.202	0.180	0.240	0.213	0.221	0.145
Appendicosporaceae	0.286	0.290	0.308	0.339	0.270	0.219	0.292	0.279	0.229	0.258	0.262	0.255	0.223	0.241
Apiosporaceae	0.235	0.232	0.278	0.306	0.256	0.198	0.269	0.233	0.196	0.264	0.273	0.272	0.204	0.207	0.253
Oxydothidaceae	0.288	0.272	0.298	0.324	0.233	0.253	0.281	0.284	0.214	0.262	0.297	0.299	0.221	0.275	0.281	0.282
Anungitiomycetaceae	0.310	0.282	0.293	0.339	0.276	0.233	0.293	0.298	0.245	0.282	0.286	0.280	0.218	0.225	0.300	0.302	0.297
Pseudosporidesmiaceae	0.316	0.314	0.308	0.335	0.302	0.265	0.292	0.321	0.280	0.263	0.257	0.258	0.298	0.293	0.297	0.313	0.318	0.269
Iodosphaeriaceae	0.356	0.329	0.363	0.369	0.348	0.275	0.333	0.318	0.298	0.270	0.321	0.306	0.252	0.271	0.341	0.330	0.320	0.316	0.258
LSU rDNA
	Beltraniaceae	Amphisphaeriaceae	Castanediellaceae	Phlogicylindriaceae	Xyladictyochaetaceae	Funiliomycetaceae	Cylindriaceae	Nothodactylariaceae	Pseudotruncatellaceae	Oxydothidaceae	Anungitiomycetaceae	Hyponectriaceae	Sporocadaceae	Clypeophysalosporaceae	Melogrammataceae	Apiosporaceae	Pseudomassariaceae	Appendicosporaceae	Pseudosporidesmiaceae	Iodosphaeriaceae
Amphisphaeriaceae	0.047
Castanediellaceae	0.041	0.058
Phlogicylindriaceae	0.055	0.056	0.050
Xyladictyochaetaceae	0.049	0.055	0.040	0.032
Funiliomycetaceae	0.059	0.061	0.044	0.050	0.042
Cylindriaceae	0.057	0.053	0.048	0.051	0.044	0.053
Nothodactylariaceae	0.064	0.067	0.055	0.052	0.040	0.040	0.047
Pseudotruncatellaceae	0.060	0.062	0.051	0.053	0.039	0.036	0.053	0.038
Oxydothidaceae	0.056	0.051	0.051	0.051	0.044	0.038	0.053	0.047	0.037
Anungitiomycetaceae	0.072	0.069	0.070	0.076	0.064	0.057	0.059	0.065	0.053	0.052
Hyponectriaceae	0.077	0.080	0.076	0.076	0.065	0.058	0.059	0.054	0.045	0.058	0.059
Sporocadaceae	0.061	0.062	0.062	0.053	0.058	0.066	0.058	0.056	0.070	0.060	0.080	0.083
Clypeophysalosporaceae	0.062	0.054	0.063	0.070	0.063	0.069	0.050	0.062	0.064	0.058	0.072	0.071	0.065
Melogrammataceae	0.060	0.056	0.059	0.047	0.037	0.044	0.046	0.044	0.045	0.036	0.054	0.058	0.055	0.053
Apiosporaceae	0.065	0.064	0.063	0.062	0.062	0.056	0.056	0.057	0.052	0.049	0.060	0.069	0.070	0.065	0.047
Pseudomassariaceae	0.067	0.068	0.064	0.071	0.063	0.065	0.063	0.065	0.065	0.062	0.080	0.076	0.073	0.068	0.065	0.073
Appendicosporaceae	0.076	0.077	0.083	0.083	0.078	0.065	0.070	0.069	0.063	0.057	0.056	0.057	0.081	0.077	0.060	0.066	0.082
Pseudosporidesmiaceae	0.088	0.093	0.098	0.097	0.086	0.078	0.092	0.086	0.077	0.066	0.066	0.080	0.100	0.092	0.073	0.079	0.102	0.053
Iodosphaeriaceae	0.087	0.091	0.098	0.102	0.096	0.084	0.104	0.100	0.087	0.073	0.074	0.089	0.110	0.098	0.084	0.089	0.106	0.067	0.051
Vialaeaceae	0.076	0.075	0.066	0.073	0.067	0.057	0.083	0.072	0.061	0.058	0.074	0.077	0.084	0.086	0.062	0.070	0.085	0.069	0.081	0.089
RPB2 rDNA
	Beltraniaceae	Melogrammataceae	Cylindriaceae	Sporocadaceae	Nothodactylariaceae	Funiliomycetaceae	Amphisphaeriaceae	Phlogicylindriaceae	Apiosporaceae	Iodosphaeriaceae	Appendicosporaceae	Oxydothidaceae
Melogrammataceae	0.208
Cylindriaceae	0.251	0.215
Sporocadaceae	0.234	0.241	0.286
Nothodactylariaceae	0.273	0.256	0.293	0.285
Funiliomycetaceae	0.252	0.259	0.270	0.309	0.262
Amphisphaeriaceae	0.259	0.247	0.285	0.256	0.303	0.304
Phlogicylindriaceae	0.262	0.229	0.271	0.296	0.296	0.309	0.313
Apiosporaceae	0.280	0.286	0.280	0.299	0.298	0.298	0.296	0.301
Iodosphaeriaceae	0.238	0.262	0.319	0.308	0.307	0.311	0.311	0.293	0.340
Appendicosporaceae	0.276	0.282	0.344	0.312	0.315	0.316	0.349	0.323	0.353	0.285
Oxydothidaceae	0.326	0.305	0.315	0.315	0.338	0.324	0.367	0.298	0.342	0.349	0.375
Anungitiomycetaceae	0.376	0.392	0.420	0.418	0.392	0.426	0.433	0.399	0.440	0.366	0.397	0.446

Table 4.. Asexual morphological features, lifestyle, host associations, and distribution of Funiliomyces species.

Species	Mycelium	Conidiophores	Conidiogenous cells	Conidia		Lifestyle	Host	Country	References
				Shape/colour	Size
Funiliomyces acaciae	2–2.5 µm, hyaline,	7–60 × 2–3.5 µm, brown, 0–7-septate	7–25 × 2–3.5 µm, brown, with flat-tipped denticles (0.5–1.5 × 0.5 µm)	narrowly fusoid ellipsoid, 2-septate, hyaline	(16–)25– 34(–37) × 2(–2.5) µm	epiphytic	Acacia koa (Fabaceae)	USA	Crous et al. (2016)
F. biseptatus	1–3 µm, hyaline to moderately brown	5–20 µm × 3–3.5 µm, moderately brown, cylindrical, 0–2-septata	cylindrical, moderately brown, with successive denticles and geniculate structure	cylindrical, 2-septata, individually, hyaline to pale smoky	(22–) 27–33 (–35) × 1.5–2 µm	saprobic	Rhododendron metternichii (Ericaceae)	Japan	Matsushima (1975)
F. biseptatus	–	–	–	–	–	saprobic	Undefined (Bromeliaceae)	Brazil	Aptroot (2004)
F. calliandrae	2–3 µm, hyphae	hyaline (appearing subhyaline with age), mostly reduced to conidiogenous cells	10–25 × 3–4 µm hyaline, prominent cylindrical denticles, 1–3 × 1.5 µm	spindle-shaped, apex subobtuse, base truncate, (3–)5–6(–8)-septate, hyaline	(37–)40–45(–47) × (2.5–)3 µm	epiphytic	Calliandra tweediei (Fabaceae)	Brazil	Crous et al. (2025)
F. fragilis	pale brown	15–30 × 4 μm at the base, 0–3 thin septa, subhyaline to pale brown	thin-walled, hyaline, slightly lobed; denticles absent; rhexolytic secession with inconspicuous scars	clavate, 2-septate, hyaline	18–26 × 1.5, base 0.6 µm wide	saprobic	Fagus sylvatica (Fagaceae)	Netherlands	de Hoog and van Oorscho (1985)
F. hwasunensis	–	6–35 × 2.2–2.8 µm， hyaline, aseptate or septate	2–2.8 μm wide，terminal, integrated, hyaline	clavate, blunt end，hyaline, 1–5 septate	10–60 × 2.2–2.8 μm	endophytic	Zoysia japonica (Poaceae)	Korea	Liu et al. (2025)
F. jiangxiensis	1.7–2.6 µm, hyaline	5–37 × 2–4 µm, hyaline, septate, sometimes reduced to conidiogenous cells	4–18 × 1–4 µm, with conspicuous, cylindrical denticles, up to 0.9 µm wide	narrowly fusoid-ellipsoid, guttulate, 0–3-septate, hyaline	20–40 × 1.5–4 µm	endophytic	Tetradium ruticarpum (Rutaceae)	China	This study
F. mavisleverae	–	–	–	–	–	epiphytic	an unidentified ornamental plant	Australia	Tan and Shivas (2024)
F. monticola	1–1.5 μm, colorless	15–40 × 2–2.5 μm, colorless, septate	12–17 × 2–3 µm, with conspicuous, truncate denticles in the apical region	fusiform, 1-septate, colourless	30–37 × 1–1.5 µm	saprobic	Andira inermis (Leguminosae)	Cuba	Castañeda and Kendr (1991)
F. retrophylli	1.5–2.5 μm, hyaline	conidiophores reduced to conidiogenous cells or with supporting cell	with 1–2 × 1 μm apex denticulate	medianly 1-septate, straight to narrowly fusoid, 6–20 × 2.5–3.5	(26–)30–33(–37) × (1.5–)2 µm	epiphytic	Retrophyllum rospigliosii (Podocarpaceae)	Colombia	Crous et al. (2022)
F. sparsus	1–1.5 μm, pale brown	0–1-septate, pale brown usually reduced to a conidiogenous cell.	7–12 × 3–4 µm, pale brown or almost colorless, with large, conspicuous, truncate denticles, 1.5–2 μm	subcylindrical, (2–)3-septated with a false septum near each end, colorless or almost colorless	26–36 × 1.5–2 µm	saprobic	decaying leaves (unidentified)	Cuba	Castañeda and Kendr (1991)
F. zapatensis	1–1.5 μm, light brown	12–60 × 1–2 µm, septate, light brown, up to 24 µm wide at apex	polyblastic, denticulate, sympodial, inflated at apex	cylindrical, 2-septata, hyaline, septa visible near extremities	18–26 × 1–1.5 µm	saprobic	Nectandra coriacea (Lauraceae)	Cuba	Castañeda Ruiz (1988)

Full text

Introduction

Dactylaria was established by Saccardo (1880) with D. purpurella as the type species, originally identified from dead leaves of Castanopsis cuspidata var. sieboldii in Japan. De Hoog (1985) expanded the circumscription of Dactylaria, recognizing 41 species classified into four sections: Dactylaria, Mirandina (G. Arnaud ex Matsush.) de Hoog, Diplorhinotrichum (Höhn.) de Hoog, and Pleurophragmium (Constantin) de Hoog. Later, Goh and Hyde (1997) expanded this framework by providing a taxonomic key for 37 additional species associated with plant material, increasing the number of recognized species to 82. Subsequent to Goh and Hyde’s (1997) review, a further 14 species have been described in Dactylaria (Paulus et al. 2003). The genus has undergone considerable taxonomic changes in recent years. Several new species have been described (Cooper 2005; Barbosa et al. 2013; Mel’nik et al. 2013; Magalhães et al. 2014; Crous et al. 2016; Crous et al. 2022; Tan and Shivas 2024; Crous et al. 2025; de Souza et al. 2025; Liu et al. 2025), while the application of molecular phylogenetic data has resulted in the transfer of some species to other genera, such as Dactylaria hyalotunicata, which is now recognized as Pseudodactylaria hyalotunicata (Crous et al. 2017). Conversely, species from other genera have been reassigned to Dactylaria based solely on traditional morphology, including Subulispora argentina and S. malaysiana (de Souza et al. 2025), although molecular evidence supporting these transfers is still lacking.

Currently, 134 epithets are listed under Dactylaria in Index Fungorum (2025) with 117 possible species listed in Species Fungorum (2025). Traditionally, members of Dactylaria are characterized by hyaline, cylindrical, rachis-bearing conidiogenous cells with apical, scattered, sympodial, tubular, or cylindrical denticles, and their conidia are hyaline, cylindrical, fusiform, or navicular, and can be unicellular or septate, with liberation occurring through schizolytic secession (de Hoog 1985; Goh and Hyde 1997; Paulus et al. 2003). Dactylaria species exhibit diverse ecological roles, with most species functioning as saprobes or parasites on a wide range of substrates including soil (Bhatt and Kendrick 2011) and plant materials (Paulus et al. 2003; Pinnoi et al. 2003; Senwanna et al. 2021; Wijesinghe et al. 2022). Additionally, some species act as endophytes that colonize living plant tissues (Wu et al. 1996), while others serve as nematode-trapping fungi (Krizková et al. 1976; Kumar and Singh 2011), plant pathogens, or opportunistic agents of animal diseases (Kralovic and Rhodes 1995; Goh and Hyde 1997).

To date, only 31 species have available sequence data in GenBank, with these data primarily corresponding to the LSU (large subunit ribosomal RNA gene) and SSU (small subunit ribosomal RNA gene) regions. No comprehensive, genus-wide phylogenetic study including the type species has been conducted and the genus’ systematic placement remains poorly resolved. Most critically, the phylogenetic position of its type species, D. purpurella, has yet to be clarified (Bussaban et al. 2005; Réblová 2009; Lin et al. 2021). LSU-based analyses conducted by Réblová (2009), which primarily addressed the sexual morph of Rhodoveronaea and the reevaluation of Pleurophragmium, demonstrated that D. purpurella, D. monticola, and D. parvispora occupy distinct lineages. The results excluded Dactylaria from synonymy with Pleurophragmium and further highlighted the polyphyletic nature of the genus. Despite its long taxonomic history, Dactylaria is taxonomically heterogeneous, leading to persistent controversy regarding its definition and classification (Goh and Hyde 1997; Paulus et al. 2003). Most species were historically established solely on morphological characteristics, particularly before the application of molecular tools (Goh and Hyde 1997; Cooper 2005; Flavia et al. 2013; Mel’nik et al. 2013; Magalhães et al. 2014). In recent years, with the advent of molecular techniques, newly described Dactylaria species have incorporated both morphological and molecular data, though typically limited to single-gene analyses. For example, D. acaciae (Crous et al. 2016), D. retrophylli (Crous et al. 2022), D. calliandrae (Crous et al. 2025), and D. hwasunensis (Liu et al. 2025) were identified using LSU sequences, whereas D. mavisleverae (Tan and Shivas 2024) was confirmed based on ITS sequences.

Interestingly, recent phylogenetic analyses of dactylaria-like taxa have revealed that Funiliomyces biseptatus consistently clusters with several “Dactylaria” species, forming a well-supported clade with uncertain taxonomic position within the order Amphisphaeriales (Crous et al. 2016, 2022, 2025). Funiliomyces biseptatus, the type and currently the only described species of the genus Funiliomyces, was established by Aptroot (2004) from a dead leaf of a Bromeliaceae plant. The genus is characterized by its fusiform ascospores with two submedian septa and furnished with both apical and median appendages. Notably, F. biseptatus represents a sexual morph, whereas the associated “Dactylaria” species are asexual forms. The close phylogenetic affinity between the sexual morph of Funiliomyces and asexual morphs of dactylaria-like taxa forms a robust, statistically well-supported clade that is phylogenetically distinct from all currently described families within Amphisphaeriales.

Amphisphaeriales was introduced by Eriksson and Hawksworth (1986). Subsequent multigene phylogenetic studies have consistently demonstrated that Amphisphaeriales and Xylariales represent distinct orders within the subclass Xylariomycetidae (Senanayake et al. 2015; Samarakoon et al. 2016; Hongsanan et al. 2017; Crous et al. 2018b). According to molecular dating analyses, the crown age of Amphisphaeriales has been estimated at around 133 MYA (Hyde et al. 2020). Members of Amphisphaeriales are characterized by gelatin-covered, interascal, tapering or reticulate paraphyses, host-immersed perithecia with or without a clypeus, hymenial asci with amyloid apices and apical rings, variable, hyaline or pigmented ascospores with variable septa, and holoblastic conidiogenesis with denticulate conidia and solitary or united conidiophores (McLaughlin and Spatafora 2014). The circumscription of Amphisphaeriales by Hyde et al. (2020) included 17 families: Amphisphaeriaceae, Appendicosporaceae, Apiosporaceae, Beltraniaceae, Castanediellaceae, Clypeophysalosporaceae, Cylindriaceae, Hyponectriaceae, Iodosphaeriaceae, Melogrammataceae, Oxydothidaceae, Phlogicylindriaceae, Pseudomassariaceae, Pseudosporidesmiaceae, Pseudotruncatellaceae, Sporocadaceae, and Xyladictyochaetaceae, while Vialaeaceae was excluded. Hyde et al. (2024) later incorporated Vialaeaceae into the order. In addition, two families originally placed in Xylariales, Nothodactylariaceae, and Anungitiomycetaceae (Crous et al. 2019b), have been transferred to Amphisphaeriales based on phylogenetic evidence (Réblová et al. 2021; Zhang et al. 2025).

Endophytic fungi associated with medicinal plants are well recognized as a rich reservoir of structurally diverse bioactive metabolites, and their interactions with host plants often contribute to the biosynthesis of pharmacologically important compounds, making them a focal point for natural product and fungal taxonomy research (Tiwari and Bae 2022; Wen et al. 2022; Gupta et al. 2023; Chandra et al. 2024). Tetradium ruticarpum (A.Juss.) Hartley, commonly known as Evodia rutaecarpa or Wu Zhu Yu in Chinese, is a medicinal plant of high value in traditional Chinese medicine (Chinese Pharmacopoeia Commission 2010). Its fruits are widely used for their pharmacological activities, including antitumor, cardiovascular protective, anti-inflammatory, antimicrobial, antioxidant, and anti-obesity effects (Li and Wang 2020; Xiao et al. 2023). Research on endophytic fungi associated with T. ruticarpum remains limited. Previously reported endophytes include Sclerotium Tode ex Fr., which produces multiple bioactive compounds (Zhu 2007), as well as other taxa identified only to the genus level, such as Cyanodermella, Guignardia, Hypoxylon, and Nigrospora (Ho et al. 2012). Two novel species, Cyphellophora guangxiensis (Mi et al. 2025b) and Pseudokeissleriella tetradii (Mi et al. 2025a), were recently reported from T. ruticarpum.

In this study, we elucidate the long-recognized polyphyly of Dactylaria**sensu lato. Phylogenetic analyses based on the LSU gene show that species historically assigned to this genus, for which LSU sequence data are available, are distributed across at least six distinct classes within Pezizomycotina, further supporting its non-monophyletic status. Phylogenetic inference using a concatenated LSU-ITS-RPB2 dataset and interfamily genetic differences recover a monophyletic lineage within Amphisphaeriales, comprising Funiliomyces biseptatus, ten species previously classified under Dactylaria, and our newly isolated strains. We thus formally propose the new family Funiliomycetaceae fam. nov. to accommodate this well-delimited clade. Additionally, we describe one new species Funiliomyces jiangxiensis from Tetradium ruticarpum, formally transfer the ten aforementioned Dactylaria species to Funiliomyces as new combinations, and present a table summarizing the asexual morphological features, lifestyle, host associations, and distribution of all known Funiliomyces species.

Materials and methods

Collection, isolation, and morphological studies

The healthy roots of Tetradium ruticarpum were collected from Jiangxi Province, China. The samples were placed into sterile self-sealing plastic bags, transported to the lab and stored at 4 °C until processing. Tissue isolation and culture method were carried out by following the method of Deng et al. (2014). The specific disinfection time was adjusted according to T. ruticarpum. The roots were repeatedly washed by using running tap water and air dried. The surface of each sample was disinfected using 75% ethanol for 1 min, and 3.3% sodium hypochlorite solution for 1 min, followed by three rinses in sterile distilled water before finally being dried on sterile filter papers. After disinfection, the roots were cut into small pieces (ca. 5 mm^2^) by a sterile scalpel and placed in Potato Dextrose Agar plates (PDA), incubated at 25 °C in darkness. When the fungi develop from the tissue segment, a few hyphal fragments were picked up and transferred to new PDA plates to obtain pure cultures.

Micromorphological characteristics from pure cultures were observed and captured using a Nikon ECLIPSE Ni-U compound microscope (Nikon Corporation, Japan), equipped with a Nikon DS-Fi3 camera. All measurements were calculated using PhotoRuler v. 1.1 software (The Genus Inocybe, Hyogo, Japan). For conidiophores and conidia, morphological data are presented as range followed by mean ± standard deviation (SD). The figures were processed using Adobe Photoshop CS6 Extended v.10.0 software (Adobe Systems, USA). The fungal cultures were deposited in the Jiangxi Agricultural University Culture Collection (JAUCC), Nanchang, China, and the China Center for Type Culture Collection (CCTCC), respectively. The dry cultures were deposited in the Herbarium of Fungi, Jiangxi Agricultural University (HFJAU), Nanchang, China. The newly introduced fungal species was submitted to the Index Fungorum (2025) and Faces of Fungi database (Jayasiri et al. 2015).

DNA extraction, PCR amplification and sequencing

Fresh mycelium was extracted using the CTAB method (Doyle and Doyle 1987). According to the locus-specific databases of the Dactylaria species (Crous et al. 2016; Crous et al. 2022), three loci (LSU, ITS, and RPB2) were chosen for polymerase chain reaction (PCR) using the primer pairs LR0R/LR5 (Vilgalys and Hester 1990), ITS1/ITS4 (White et al. 1990), and fRPB2-5F/fRPB2-7CR (Liu et al. 1999), respectively. The final volume of the PCR reaction mixture was 25 μL, contained 12.5 μL 2×Taq PCR MasterMix (Qingke, Changsha, China), 1 μL each forward and reverse primer (0.2 μM), 1 μL template DNA (circa 50–100 ng), and 9.5 μL ddH_2_O. For ITS and LSU genes, the PCR parameters were initial denaturation 94 °C for 5 min; followed by 35 cycles consisting of denaturation at 94 °C for 30 seconds, annealing at 55 °C for 50 seconds and extension at 72 °C for 60 seconds; and final extension at 72 °C for 10 min. For RPB2 gene, the PCR parameters were initial denaturation 94 °C for 5 min; followed by 40 cycles consisting of denaturation at 94 °C for 60 seconds, annealing at 58 °C for 60 seconds and extension at 72 °C for 60 seconds; and a final extension at 72 °C for 10 min. The PCR products were purified and sequenced by QingKe Biotechnology Co. (Changsha, China). All sequences were assembled using BioEdit v.7.2.6 (Hall 1999). The new sequences were deposited in GenBank (https://www.ncbi.nlm.nih.gov/). The sequence alignments and resulting phylogenetic trees have been deposited in TreeBASE (www.treebase.org).

Reference datasets

We combined the new ribosomal and protein coding sequences with data from the National Center for Biotechnology Information (NCBI). Accession numbers for all sequences used for the molecular analyses are provided in Tables 1, 2.

1. Pezizomycotina: To determine the phylogenetic placement of Dactylaria within Pezizomycotina, we compiled a dataset representing all major ascomycete classes and conducted LSU single-gene analyses (Beimforde et al. 2020). Of the 31 Dactylaria species with molecular data in GenBank, five (D. acerosa, D. appendiculata, D. candidula, D. junci, and D. lanosa) lack LSU sequences and were excluded. The final dataset included 124 species, comprising 26 Dactyla­ria species. Accession numbers for all sequences are provided in Table 1.

2. Amphisphaeriales: To assess the placement of our two new strains, 71 taxa were selected based on the Pezizomycotina phylogeny of Dactylaria and recent publications (Crous et al. 2016, 2022, 2025; Tan and Shivas 2024; Liu et al. 2025). Phylogenetic analyses of Dactylaria species affiliated with Amphisphaeriales were performed using LSU, ITS, and RPB2 sequences. Accession numbers for all sequences are listed in Table 2.

Phylogenetic analyses

Phylogenetic analyses were conducted using Maximum Likelihood (ML) and Bayesian Inference (BI). Sequences for both datasets were obtained from GenBank (http://www.ncbi.nlm.nih.gov/). All sequences were aligned with the online version of MAFFT v. 7 (https://mafft.cbrc.jp/alignment/server/) (Katoh and Standley 2013) and manually refined in BioEdit v. 7.2.6 (Hall 1999), with gaps treated as missing data. Dataset 1 contained only LSU sequences, whereas for Dataset 2, LSU, ITS, and RPB2 sequences were concatenated into a supermatrix using PhyloSuite v. 1.2.2 (Zhang et al. 2020). Alignment file formats were subsequently converted with ALTER (Alignment Transformation Environment; http://www.sing-group.org/ALTER/).

ML analyses were carried out on the dataset using IQ-TREE v.1.6 (http://iqtree.cibiv.univie.ac.at/) with partitioning, and the optimal substitution model for each partition was determined by IQ-TREE. (Trifinopoulos et al. 2016). Clade support for the ML analyses was assessed using the Shimodaira-Hasegawa-like approximate likelihood ratio test (SH-aLRT) with 1,000 replicates (Guindon et al. 2010) and 1,000 replicates of the ultrafast bootstrap (UFB) (Hoang et al. 2017). Nodes with support values of both SH-aLRT ≥ 80 and UFB ≥ 95 were considered supported, nodes with one of SH-aLRT ≥ 80 or UFB ≥ 95 were weakly supported, and nodes with both SH-aLRT < 80 and UFB < 95 were unsupported, and other parameters were used for the default settings (Liu et al. 2022).

Bayesian Inference (BI) analyses were performed on the CIPRES Science Gateway (http://www.phylo.org/) (Miller et al. 2010) using MrBayes 3.2.7 (Ronquist et al. 2012). For both datasets, each gene partition was analyzed under the GTR+I+G model, which was selected as the best-fit model using MrModeltest 2.3 in PAUP* v4.0b (Swofford 2002; Nylander 2004). Two independent Markov Chain Monte Carlo (MCMC) runs were conducted for 10,000,000 generations, sampling every 1,000 generations, with the first 25% of samples discarded as burn-in. Convergence was assessed by ensuring the average standard deviation of split frequencies (ASDSF) fell below 0.01. Bayesian posterior probabilities (BPP) were used to assess node support (Rannala and Yang 1996). Resulting trees were visualized in FigTree v. 1.4.4 (Rambaut 2018) and edited using the online service tvBOT (https://www.chiplot.online/tvbot.html) (Xie et al. 2023). Genetic distances were estimated using the Kimura 2-parameter (K2P) model in MEGA-X (Kumar et al. 2018) based on ITS, LSU, and RPB2 sequences of species representing different family within Amphisphaeriales.

Results

Molecular analysis

Based on a nucleotide BLAST search in GenBank database using the LSU sequence, the closest matches are Dactylaria species, including Funiliomyces hwasunensis (≡ D. calliandrae) CMML 20-35 [GenBank PQ741487; identities = 820/824 (99%), gaps = 0/824 (0%)], F. calliandrae (≡D. calliandrae) CPC 48004 [GenBank PV664963; identities = 805/811 (99%), gaps = 0/811 (0%)], F. fragilis (≡ D. fragilis) P057 [GenBank EU107290; identities = 798/807 (99%), gaps = 0/811 (0%)]. The closest match for the ITS sequence is Fusidium griseum Trtsf08 [GenBank GU479905; identities = 468/495 (95%), gaps = 7/495 (1%)], D. acerosa ICMP 13178 [GenBank OR543730; identities = 465/494 (94%), gaps = 4/494 (4%)], and F. calliandrae (≡ D. calliandrae) CPC 48004 [GenBank PV664937; identities = 454/479 (95%), gaps = 6/479 (4%)]. For the RPB2 sequence, the closest matches included Dicyma funiculosa CBS 323.86 [GenBank KU684306; identities = 579/735 (79%), gaps = 6/735 (0%)], and Xylaria liquidambaris FCATAS879 [GenBank MZ707110; identities = 553/700 (79%), gaps = 8/700 (1%)]. This situation may be due to the limited availability of RPB2 gene sequences for this genus.

Phylogenetic analysis

The phylogenetic hypothesis inferred from the LSU gene dataset for Pezizomycotina is presented in Fig. 1, with Candida albicans (ATCC 18804) and Saccharomyces cerevisiae (NRRL Y-12632) selected as outgroup taxa to root the tree, following previous large-scale phylogenetic studies of Pezizomycotina (Beimforde et al. 2020). The LSU alignment comprised 898 bp (including gaps), with 445 parsimony-informative sites, 94 singletons, and 359 constant sites. Maximum likelihood (ML) analysis recovered 629 distinct alignment patterns, with 7.33% undetermined characters or gaps. The best-scoring ML tree (–lnL = 22860.942) is shown in Fig. 1. Overall, the topologies obtained from ML and Bayesian inference (BI) analyses were highly congruent, differing only in the placement of a few taxa and in several unresolved clades. In this analysis, twenty-two Dactylaria species are distributed across six different classes within Pezizomycotina, namely Orbiliomycetes (one species), Leotiomycetes (one species), Eurotiomycetes (two species), Sordariomycetes (16 species), Dothideomycetes (one species), and Lecanoromycetes (one species). The remaining four species could not be confidently assigned to any currently recognized class of Pezizomycotina. The phylogenetic tree reveals that Dactylaria is polyphyletic, with most species clustering in Sordariomycetes. The exact taxonomic position of the type species, D. purpurella, remains uncertain. It forms a distinct lineage with D. humicola and D. dimorpha, representing an incertae sedis lineage in Pezizomycotina. In addition, our newly isolated strain clustered in the Sordariomycetes clade with ten species of Funiliomyces (formerly treated as Dactylaria) in Funiliomycetaceae fam. nov. with branch support values of SH-aLRT = 87.4%, UFB = 94%, and BPP = 0.88 (UFB value < 95% and BPP < 0.90 are not shown on the tree) (Fig. 1).

Maximum likelihood phylogenetic tree of Pezizomycotina based on LSU sequences, with Shimodaira-Hasegawa-like approximate likelihood ratio test (SH-aLRT) (left), ultrafast bootstrap (UFB) (middle), and Bayesian posterior probabilities (BPP) values (right) near the corresponding node. Only one of SH-aLRT > 80 or UFB > 95 for ML and BYPP > 0.90 for BI is indicated along the branches (SH-aLRT/UFB/BPP). The “Dactylaria” species are indicated in red, the newly generated sequences are indicated in green, and species for reclassification from Dactylaria to Funiliomyces are in blue. Ex-type strains are marked with T after the strain number. The ML phylogram is available in TreeBASE (study accession S32460; http://purl.org/phylo/treebase/phylows/study/TB2:S32460).

The phylogenetic tree of Amphisphaeriales based on the three-gene dataset, rooted with Achaetomium macrosporum (CBS 532.94), Chaetomium elatum (CBS 374.66), and Sordaria fimicola (CBS 723.96), is shown in Fig. 2. The combined dataset of LSU, ITS, and RPB2 consisted of 2642 bp including gaps (LSU 857 bp, ITS 728 bp, and RPB2 1057 bp), which contained 1501 parsimony-informative characters, 247 singleton sites, and 1339 constant characters. The maximum likelihood matrix had 1804 distinct alignment patterns with 37.19% undetermined characters or gaps. The best maximum likelihood tree, with a final likelihood value of -29373.458, is shown in Fig. 2. The ML tree and BI analysis tree were similar, except for some interchanged positions and some unseparated clade. In this study, phylogenetic analyses revealed that our two new strains, together with ten previously described “Dactylaria” species and one Funiliomyces species, formed a distinct yet unplaced lineage within Amphisphaeriales (Fig. 2). The overall tree topology (Fig. 2) recovered in this study is largely congruent with those reported by Crous et al. (2022, 2025). Our phylogenetic analyses further confirmed that Dactylaria sensu lato is polyphyletic. Dactylaria purpurella, the type species of the genus, forms an independent lineage within Pezizomycotina and is distantly related to the Amphisphaeriales clade comprising ten “Dactylaria” taxa. (Fig. 1). Accordingly, these species in Amphisphaeriales are not congeneric with the type of Dactylaria. Based on phylogenetic evidence and following the principles of monophyly and the “One Fungus, One Name” concept, these ten species are here transferred to the genus Funiliomyces. Although Funiliomyces biseptatus is only known from its sexual morph, whereas the transferred species are asexual, phylogenetic placement indicates that they represent different morphs of the same genus. In addition, our analyses also showed that the two newly generated strains, CCTCC AF 2023033 and CCTCC AF 2023034, form a distinct branch and cluster as sister lineages with Funiliomyces hwasunensis (≡ Dactylaria hwasunensis) and F. calliandrae (≡ D. calliandrae), supported by SH-aLRT = 98.8%, UFB = 100%, and Bayesian posterior probability = 0.98 (Fig. 2), indicating that they represent a phylogenetically novel species in Funiliomyces.

Maximum likelihood phylogenetic tree of Amphisphaeriales based on combined LSU, ITS, and RPB2 sequences, with Shimodaira-Hasegawa-like approximate likelihood ratio test (SH-aLRT) (left), ultrafast bootstrap (UFB) (middle), and Bayesian posterior probabilities (BPP) values (right) near the corresponding node. Only one of SH-aLRT > 80 or UFB > 95 for ML and BYPP > 0.90 for BI is indicated along the branches (SH-aLRT/UFB/BPP). Ex-type strains are marked with T after the strain number. The newly generated sequences are indicated in red and species for reclassification are in blue. The ML phylogram is available in TreeBASE (study accession S32462; http://purl.org/phylo/treebase/phylows/study/TB2:S32462).

Furthermore, the phylogenetic tree inferred from the combined three-locus dataset resolved twenty main clades within Amphisphaeriales (Fig. 2), corresponding to the families Beltraniaceae (98.8\100\1.0), Amphisphaeriaceae (100\100\1.0), Castanediellaceae (96\100\1.0), Sporocadaceae (100\100\1.0), Phlogicylindriaceae (99.6\100\1.0), Xyladictyochaetaceae (-\95\0.98), Clypeophysalosporaceae (99.7\100\1.0), Cylindriaceae (100\100\1.0), Pseudomassariaceae (98.2\99\1.0), Apiosporaceae (100\100\1.0), Melogrammataceae (100\100\1.0), Pseudotruncatellaceae (100\100\1.0), a distinct clade formed by Funiliomyces (86.6--), Nothodactylariaceae (96.1\98\1.0), Oxydothidaceae (100\100\1.0), Appendicosporaceae (92.8\99\1.0), Hyponectriaceae (-/-/-), Anungitiomycetaceae (100\100\1.0), Vialaeaceae (100\100\1.0), Pseudosporidesmiaceae (100\100\1.0), and Iodosphaeriaceae (100\100\1.0). Both Bayesian inference and maximum likelihood analyses supported the monophyly of this Funiliomyces clade and its sister relationship to Nothodactylariaceae (SH-aLRT = 99.9%, UFB = 94%, BPP = 0.99; UFB < 95% is not shown on the tree). Based on these phylogenetic results, a new family, Funiliomycetaceae fam. nov., is proposed to accommodate the Funiliomyces lineage.

Genetic distance analysis among families within Amphisphaeriales

To further clarify the phylogenetic position of the “Dactylaria” lineage within Amphisphaeriales, we conducted genetic divergence analyses based on ITS, LSU, and RPB2 sequences.

Within Amphisphaeriales, the ITS genetic distances among 19 families range from 0.110 (between Sporocadaceae and Beltraniaceae) to 0.369 (between Pseudosporidesmiaceae and Iodosphaeriaceae), indicating substantial inter-family variation in this locus (Table 3). When focusing on Funiliomycetaceae, its ITS genetic distances to the other 19 families range from 0.156 to 0.306, with the smallest distance to Nothodactylariaceae (0.156) and the largest to Iodosphaeriaceae (0.306). Notably, the minimum divergence of Funiliomycetaceae from other families (0.156) is still greater than the smallest inter-family distance within the order (0.110), suggesting that Funiliomycetaceae is not closely allied with any established family.

For the LSU dataset, the genetic distances among the 20 families range from 0.032 between Xyladictyochaetaceae and Phlogicylindriaceae to 0.110 between Sporocadaceae and Iodosphaeriaceae. The LSU distances between Funiliomycetaceae and the other families vary from 0.036 with Pseudotruncatellaceae to 0.084 with Iodosphaeriaceae. Again, the smallest LSU distance between Funiliomycetaceae and Pseudotruncatellaceae (0.036) remains higher than the overall minimum value (0.032) observed within the order, supporting its independent molecular identity.

For the RPB2 dataset, only 13 families were included because not all families within Amphisphaeriales have publicly available RPB2 sequences. Nevertheless, the inter-family distances reveal clear divergence of Funiliomycetaceae. Overall, the inter-family distances range from 0.208 between Melogrammataceae and Beltraniaceae to 0.446 between Anungitiomycetaceae and Oxydothidaceae. The RPB2 distances between Funiliomycetaceae and the other families range from 0.252 with Beltraniaceae to 0.426 with Anungitiomycetaceae. The minimum distance between Funiliomycetaceae and Beltraniaceae (0.252) is notably higher than the smallest inter-family distance in the dataset (0.208), indicating consistent divergence.

Collectively, the analyses based on ITS, LSU, and RPB2 sequences demonstrate that Funiliomycetaceae is genetically distinct from all other families within Amphisphaeriales. Across three loci, its inter-family distances are consistently greater than the smallest distances observed among other established families, providing robust molecular evidence that Funiliomycetaceae represents an independent family within the order.

Taxonomy

Funiliomycetaceae

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

L.X. Mi, K.D. Hyde, H.Y. Song & D.M. Hu fam. nov.

1F5906FE-B7AA-56E9-BDE3-BAAB5CA80D61

Index Fungorum: IF904529

Facesoffungi Number: FoF18896

Etymology.

Name refers to the type genus Funiliomyces.

Type genus.

Funiliomyces Aptroot, Studies in Mycology 50(2): 309 (2004).

Description.

Saprobic, endophytic, or epiphytic on diverse plant hosts in tropical to temperate regions. Sexual morph: Ascomata black, subglobose, immersed to erumpent. Wall composed of irregular layers of regularly melanized, flattened cells, with no color change in KOH. Paraphyses absent. Asci cylindrical, 8-spored, with a thickened apex bearing a central refractive, IKI-negative apical apparatus, and enclosed by parenchymatous tissue. Ascospores pale brown, torpedo-shaped, 2-septate, upper cell pointed, lower cell rounded, hyaline granules or oil droplets, bearing two hyaline mucilaginous appendages. Asexual morph: Mycelium consisting of branched, septate, smooth, hyaline to pale brown hyphae, sometimes forming hyphal ropes. Conidiophores macronematous, mononematous, solitary or in small groups, erect, straight to flexuous, subcylindrical to cylindrical (apex sometimes inflated), simple or occasionally branched, hyaline to brown, septate, sometimes reduced to conidiogenous cells. Conidiogenous cells integrated, terminal or lateral, sympodial, mono- to polyblastic, hyaline to brown, cylindrical to clavate, with flat-tipped or denticulate apices; denticles (when present) large, cylindrical to geniculate, cylindrical, truncate, or pimple-like, or lacking entirely. Conidia solitary, hyaline to pale smoky, smooth, septate, narrowly fusiform to cylindrical, with obtuse, subobtuse, or tapering apices and truncate or rounded bases; dimensions variable among species.

Notes.

A new family, Funiliomycetaceae, is proposed to accommodate a distinct, strongly supported clade comprising the genus Funiliomyces and several related lineages historically identified as “Dactylaria”. Funiliomycetaceae shares general sexual characters with Amphisphaeriales, including immersed to semi-immersed ascomata and unitunicate asci with an apical ring (Aptroot 2004). However, the family is readily distinguished from all currently accepted families in the order by its unique ascospore morphology: the ascospores are fusiform, consistently biseptate, and bear both apical and median appendages, a combination of characters not reported in other families of Amphisphaeriales (Aptroot 2004). The asexual morph of Funiliomycetaceae is hyphomycetous, characterized by conidiogenous cells that are sympodial, terminal or lateral, integrated, and denticulate (when present, denticles are large and cylindrical to geniculate), producing solitary, hyaline to pale smoky, smooth, septate, narrowly fusiform to cylindrical conidia with obtuse to tapering apices and truncate or rounded bases. Funiliomycetaceae shares similar sexual features with Nothodactylariaceae (Nothodactylaria), such as Nothodactylaria nephrolepidis (Cours et al. 2019b). However, Funiliomycetaceae is phylogenetically distinct from the families in Amphisphaeriales, as supported by multi-locus phylogenetic analyses (Fig. 2) and inter-familial genetic distance comparisons (Table 3).

Funiliomyces

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

Aptroot, Stud. Mycol. 50 (2): 309 (2004)

8987BC25-26FA-557A-A755-8B01ACE7817F

Index Fungorum: IF500077

Facesoffungi Number: FoF18897

Type species.

Funiliomyces biseptatus Aptroot, Stud. Mycol. 50 (2): 309 (2004). Index Fungorum: IF500164.

Holotype.

CBS H-10505.

Type information.

Brazil, Minas Gerais, Catas Altas, Serro do Caraça, Parque Natural do Caraça, near Funil, 1 km NW of monastery Santuário do Caraça, 20°06'S, 43°29'W, on dead leaf of Bromeliaceae in rock field, 18 Sept. 1997, A. Aptroot, holotype herb. CBS H-10505, isotypes herb. SP, living culture ex-type CBS 100373, also dried culture CBS H-10506.

Description.

See Aptroot (2004) on page 309.

Emended diagnosis.

Saprobic, endophytic, or epiphytic on diverse plant hosts in tropical to temperate regions. Sexual morph: Ascomata black, subglobose, immersed to erumpent. Wall composed of irregular layers of regularly melanized, flattened cells, with no color change in KOH. Paraphyses absent. Asci cylindrical, 8-spored, with a thickened apex bearing a central refractive, IKI-negative apical apparatus, and enclosed by parenchymatous tissue. Ascospores pale brown, torpedo-shaped, 2-septate, upper cell pointed, lower cell rounded, hyaline granules or oil droplets, bearing two hyaline mucilaginous appendages. Asexual morph: Mycelium consisting of branched, septate, smooth, hyaline to pale brown hyphae, sometimes forming hyphal ropes. Conidiophores macronematous, mononematous, solitary or in small groups, erect, straight to flexuous, subcylindrical to cylindrical (apex sometimes inflated), simple or occasionally branched, hyaline to brown, septate, sometimes reduced to conidiogenous cells. Conidiogenous cells integrated, terminal or lateral, sympodial, mono- to polyblastic, hyaline to brown, cylindrical to clavate, with flat-tipped or denticulate apices; denticles (when present) large, cylindrical to geniculate, cylindrical, truncate, or pimple-like, or lacking entirely. Conidia solitary, hyaline to pale smoky, smooth, septate, narrowly fusiform to cylindrical, with obtuse, subobtuse, or tapering apices and truncate or rounded bases, dimensions variable among species.

Notes.

Funiliomyces was first described by Aptroot (2004) as a monospecific genus, containing only its type species F. biseptatuss. With respect to ten species previously assigned to Dactylaria, our phylogenetic evidence shows they are phylogenetically distant from the type species of the genus (Dactylaria purpurella) (Fig. 1). However, these ten species cluster with Funiliomyces biseptatus, forming a distinct, well-supported lineage (Fig. 2). Therefore, in accordance with the “One Fungus, One Name” principle, we transfer these ten species to Funiliomyces and introduce the new species F. jiangxiensis within this genus. This emendation expands the genus to encompass both sexual and asexual morphs. The type species, F. biseptatus, represents the sexual morph, characterized by torpedo-shaped ascospores with two nearly central septa and appendages. In contrast, the eleven other species represent the asexual morphs, producing hyaline, septate conidiophores with sympodial, denticulate conidiogenous cells and solitary, hyaline, clavate or fusoid-ellipsoid conidia.

Funiliomyces

jiangxiensis

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

L.X. Mi, K.D. Hyde, H.Y. Song & D.M. Hu sp. nov

C4E7F4E0-6D94-56B2-B84C-E02A364C46B2

Index Fungorum: IF904530

Facesoffungi Number: FoF18898

Fig. 3

Etymology.

The name refers to the place where the fungal was collected.

Funiliomyces jiangxiensis sp. nov. (HFJAU10125, holotype) a, b Colonies on the front and back of PDA medium (for 5 days); c Hyphae and sporulation structures; d, e Conidiophores bearing a conidium initial on one of its denticles; f, g Conidiophores; h–l Conidia. Scale bars: 10 μm (c, d); 5 μm (e); 10 μm (f, g); 20 μm (h); 10 μm (i–l).

Holotype.

HFJAU10125.

Description.

Endophytic fungus isolated from the roots of Tetradium ruticarpum. Sexual morph: Undetermined. Asexual morph: Mycelium consisted of hyaline, smooth, branched, septate, 1.7–2.6 µm diam hyphae. Conidiophores (6–)8.5–17.5(–23.5) × 1.8–3.5(–4) µm (mean ± SD = 12.5 ± 4.3 × 3 ± 0.4 µm, n = 35), macronematous, mononematous, hyaline, subcylindrical, arising from terminal or intercalary parts of aerial hyphae, mostly reduced to conidiogenous cells (rarely with a supporting cell), tapering towards the base, apex polyblastic, sympodial, inflated or geniculous-sinuous, with conspicuous, cylindrical denticles, up to 0.9 µm wide. Conidia 20–40 × 1.5–3.5 μm (mean ± SD = 27.8 ± 3.9 × 2.6 ± 0.3 µm, n = 40), hyaline, narrowly fusoid-ellipsoid, 0–3-septate, guttulate, apex sub-obtuse, base truncate.

Culture characteristics.

Colonies on PDA at 25 °C for 5 days, convex, white with cream margin, reverse pale brown with cream margin, no pigment in agar.

Material examined.

CHINA, Jiangxi Province, Ganzhou City, Ganxian District, 25.6121°N, 115.1211°E 412.9 m asl, isolated from healthy roots of Tetradium ruticarpum, 26 June 2022, Lixue Mi, dry culture HFJAU10125 (holotype); ex-type JAUCC 5298 = CCTCC AF 2023034; ibid., Yichun City, Zhangshu county-level city, 27.9931°N, 115.2123°E, 46 m asl, 21 September 2021, Lixue Mi, dry culture HFJAU10124; living culture: JAUCC 4255 = CCTCC AF 2023033.

Notes.

In the multi-gene analysis, Funiliomyces jiangxiensis (strains CCTCC AF 2023033 and CCTCC AF 2023034) forms a distinct lineage that groups with F. hwasunensis (≡ Dactylaria hwasunensis) and F. calliandrae (≡ D. calliandrae) as a sister branch, supported by high statistical values in the phylogenetic tree (SH-aLRT/UFB/BPP = 98.8/100/0.98). Morphologically, Funiliomyces jiangxiensis can be distinguished from F. hwasunensis by their conidiogenous cells and conidia. Specifically, Funiliomyces jiangxiensis develops distinct denticles on its conidiogenous cells, a feature that is absent in F. hwasunensis. Additionally, there is a notable difference in the number of septa in their conidia: Funiliomyces jiangxiensis has conidia with 0–3 septa, whereas F. hwasunensis produces conidia with more septa, ranging from 1 to 5. In terms of morphology, F. jiangxiensis resembles F. calliandrae but can be distinguished from it. Both have denticles on conidiogenous cells, yet those of F. jiangxiensis (0.9 μm) are shorter than F. calliandrae’s (1–3 μm). Additionally, the conidia of F. jiangxiensis (20–40 μm) are shorter than those of F. calliandrae [(37–)40–45(–47) μm]. The species was isolated from the roots of Tetradium ruticarpum (medicinal plant), which provides a basis for further research on its potential functional properties.

Other species in Funiliomyces

Funiliomyces

acaciae

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(Crous) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

F81157E9-1057-56E8-A44A-9566BD4A5A14

Index Fungorum: IF819073

Basionym.

Dactylaria acaciae Crous, Persoonia 37: 321 (2016).

Holotype.

CBS H-22876.

Type information.

USA, Hawaii, Oahu, on leaves of Acacia koa (Fabaceae), 30 September 2015, J.J. Le Roux (holotype CBS H-22876, culture ex-type CPC 29771 = CBS 142087).

Description.

See the original description in D’Souza et al. (2002) on page 141.

Funiliomyces

bisepatus

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(Matsushima) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

35065E27-80F5-5BFF-A470-B6A5F948E900

Index Fungorum: IF312614

Basionym.

Dactylaria biseptata Matsushima, Icones Microfungorum a Matsushima lectorum: 48 (1975).

Holotype.

MFC-4029.

Type information.

Japan, Ohdaigahara, Nara Pref, on a rotten leaf of Rhododendron metternichii (Ericaceae), July 1970, MFC-4029 (holotype).

Description.

See the original description in Matsushima (1975) on page 48–49.

Funiliomyces

calliandrae

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(Crous) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

49F7F1F4-0A9A-57DD-8C14-DADE48E5762E

Index Fungorum: IF859210

Basionym.

Dactylaria calliandrae Crous et al. Persoonia 54: 376–377 (2025).

Holotype.

CBS H-25715.

Type information.

Brazil, Minas Gerais, Viçosa, Clonar nursery, on living leaf of Calliandra tweediei (Fabaceae), 25 February, 2024, P.W. Crous, HPC 4399 (holotype CBS H-25715; culture ex-type COAD 3994 = CPC 48004).

Description.

See the original description in Crous et al. (2025) on page 376–377.

Funiliomyces

fragilis

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(de Hoog) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

BEA3A81B-7FCA-508A-BFFB-06B2C5C603EE

Index Fungorum: IF104169

Basionym.

Dactylaria fragilis de Hoog, Studies in Mycology 26: 30 (1985).

Holotype.

No.6074(CBS).

Type information.

The Netherlands, Opsterland, Oldeterp, on cupules of Fagus sylvatica (Fagaceae), H.A. van der Aa, October, 1977.

Description.

See the original description in de Hoog and van Oorscho (1985) on page 30.

Funiliomyces

hwasunensis

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(H.F. Liu & H.K. Sang) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

0E0930FD-B913-5AEF-BDA3-199EF4DB63F0

Index Fungorum: IF857258

Basionym.

Dactylaria hwasunensis H.F. Liu & H.K, IMA Fungus 16(e138479): 10 (2025).

Holotype.

CMML 20-35H.

Type information.

Korea, South Jeolla Province, Hwasun, isolated from roots of Zoysia japonica (Poaceae), October 2020, H. Liu & H. Sang, holotype CMML 20-35H, ex-holotype CMML 20-35, ex-isotype CMML 20-88.

Description.

See the original description in Liu et al. (2025) on page 12–14.

Funiliomyces

mavisleverae

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(Y.P. Tan, Bishop-Hurley & Marney) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

C0E4BDBC-B516-562F-AB90-5A208DABF199

Index Fungorum: IF902836

Basionym.

Dactylaria mavisleverae Y.P. Tan, Bishop-Hurley & Marney, Index of Australian Fungi 46: 3 (2024).

Holotype.

BRIP 76362a.

Type information.

Australia, Queensland, Brisbane, phylloplane of unidentified ornamental plant, January, 2024, T.S. Marney, BRIP 76362a (holotype).

Description.

See the original description in Tan et al. (2024) on page 3–4.

Funiliomyces

monticola

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(R. F. Castañeda & W. B. Kendr.) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

7E02CE3E-BF51-5EC0-A89D-F02181465D82

Index Fungorum: IF361523

Basionym.

Dactylaria monticola R.F. Castañeda & W.B. Kendr, University of Waterloo Biology Series, 35: 30 (1991).

Holotype.

INIFAT C 91/82.

Type information.

Cuba, Granma, Buey Arriba, La Estrella, on dead leaves of Andira inermis (Leguminosae), R.F. Castañeda, 14 March 1991.

Description.

See the original description in Castañeda and Kendr (1991) on page 30.

Funiliomyces

retrophylli

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(Crous) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

D277145E-E22D-5B20-8EEC-CAA7314CC75B

Index Fungorum: IF844283

Basionym.

Dactylaria retrophylli Crous, Fungal Systematics and Evolution 10: 41 (2022).

Holotype.

HPC 3260.

Type information.

Colombia, Finca El Cedral, on leaves of Retrophyllum rospigliosii (Podocarpaceae), M.J. Wingfield, February, 2020, HPC 3260 (holotype CBS H-24817, culture ex-type CPC 39510 = CBS 148271).

Description.

See the original description in Crous et al. (2022) on page 41–42.

Funiliomyces

sparsus

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(R. F. Castañeda & W. B. Kendr.) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

0F863F0C-362F-5604-BC23-01C7C748F899

Index Fungorum: IF361528

Basionym.

Dactylaria sparsa R.F. Castañeda & W.B. Kendr, University of Waterloo Biology Series. 35:33 (1991).

Holotype.

INIFAT C 91/68-2.

Type information.

Cuba, C. Habana, Santiago de las Vegas, on decaying leaves, R.F. Castañeda, 18 February 1990.

Description.

See the original description in Castañeda and Kendr (1991) on page 33.

Funiliomyces

zapatensis

Taxon classification

Animalia

AmphisphaerialesFuniliomycetaceae

(R.F. Castañeda) L.X. Mi, H.Y. Song, D.M. Hu & K.D. Hyde comb. nov.

8E847D83-3E72-582C-818E-5CFA62B73CD2

Index Fungorum: IF125340

Basionym.

Dactylaria zapatensis R.F. Castañeda, Fungi Cubenses III (La Habana): 5 (1988)

Holotype.

INIFAT C85/98

Type information.

Cuba, Matanzas, Ciénaga de Zapata, on fallen leaves of Nectandra coriacea (Lauraceae), R.F. Castañeda Ruiz, 26 May 1985.

Description.

See the original description in Castañeda Ruiz (1988) on page 5.

Funiliomyces currently includes only a single sexual morph species, F. biseptatus; therefore, we summarized the asexual morphological features, lifestyle, host associations, and distribution for all species (Table 4) to facilitate comparison within the genus.

Discussion

Previous studies have consistently demonstrated that Dactylaria is polyphyletic, yet the taxonomic placement of its type species, D. purpurella, has never been formally resolved. To date, there has been no phylogenetic study specifically designed to address the classification of D. purpurella; rather, it has generally been included only as part of broader datasets, without being treated as a focal taxon. As a result, its placement has remained ambiguous. For instance, Bussaban et al. (2005) showed that D. purpurella clustered with Ochroconis rather than other Dactylaria species based on ITS sequence data. Similarly, Réblová (2009) using LSU sequence data, demonstrated that D. purpurella, D. monticola, and D. parvispora fall into distinct clades, with D. purpurella forming an isolated lineage, while D. monticola grouping within Xylariales, and D. parvispora clustering with Papulosa amerospora in Papulosaceae. Subsequent studies further added to this complexity, placing D. purpurella within Magnaporthales (Klaubauf et al. 2014) or Venturiales (Lin et al. 2021). Taken together, these conflicting results highlight not only the unstable phylogenetic position of D. purpurella, but also the broader taxonomic challenges in delimiting generic boundaries within Dactylaria. In our study, the LSU-based phylogenetic analysis of Pezizomycotina demonstrated that Dactylaria is polyphyletic, with its species distributed across multiple classes, predominantly within Sordariomycetes. However, the phylogenetic placement of four species, D. purpurella (the type species), D. humicola, D. dimorpha, and D. dimorphospora, has not been assigned to any of the known classes within Pezizomycotina and thus remains unresolved (Fig. 1). As the first systematic investigation of the phylogenetic position of Dactylaria, based on all available LSU sequences data, our work provides a comprehensive overview of its taxonomic placement. The results emphasize the necessity of additional sampling and multi-locus phylogenetic analyses to resolve the taxonomy and evolutionary relationships within the genus, thereby laying the foundation for future refinement of its classification.

Although Dactylaria is polyphyletic, new species have continued to be introduced into the genus in recent years (Crous et al. 2016, 2022, 2025), all of which are placed within Amphisphaeriales, forming a single unresolved lineage. Fortunately, we also collected two new strains that cluster with the recently published Dactylaria species. Phylogenetic analysis based on the combined LSU, ITS, and RPB2 loci reveals that ten previously described “Dactylaria” species, our newly obtained isolates, and Funiliomyces biseptatus (type strain CBS 100373) cluster together into a single clade within Amphisphaeriales (Fig. 2). Topologically, this clade is comparable to lineages recovered in multiple previous studies, which remained unplaced within any established family of Amphisphaeriales (Crous et al. 2016, 2022, 2025). With the discovery of additional species of Nothodactylariaceae (Zhang et al. 2025) and their incorporation into the phylogenetic analyses of this study, this well-distinguished clade is recovered as a sister lineage to Nothodactylaria, the sole genus currently recognized in Nothodactylariaceae, with strong phylogenetic support (SH-aLRT/UFB/BPP = 99.9/94/0.99). In addition, multi-locus genetic distance analyses (ITS, LSU, and RPB2) revealed substantial divergence between this clade and other families within Amphisphaeriales, further supporting its independence from previously recognized families. Based on our results (Fig. 2, Table 3), both the phylogenetic analyses and genetic distance divergence clearly support that this distinct lineage represents a family-level taxon within Amphisphaeriales. Therefore, we establish the new family Funiliomycetaceae within Amphisphaeriales to accommodate this distinct lineage.

Morphologically, Funiliomycetaceae integrates both sexual and asexual morphs. Funiliomyces biseptatus, the only species with a sexual morph, produces torpedo-shaped ascospores with two nearly central septa and appendages, while the eleven other species represent asexual morphs, producing hyaline, septate conidiophores with sympodial, denticulate conidiogenous cells and solitary, hyaline, clavate or fusoid-ellipsoid conidia. This integration of sexual and asexual morphs within a single lineage highlights the evolutionary significance of the family and provides a robust morphological framework for family-level delimitation. When compared with Nothodactylariaceae, morphological evidence corroborates the close relationship between the two families, yet the overall conidial range in Funiliomycetaceae is considerably broader, encompassing species with distinctly larger conidia (Table 4). A more definitive diagnostic feature is the diversity of conidiogenous cells, which bear large cylindrical to geniculate denticles or lack denticles entirely, in sharp contrast to the uniform, pimple-like denticles of Nothodactylariaceae (Crous et al. 2019b, Zhang et al. 2025). Future studies will require additional species sampling and isolation to uncover sexual morphs, explore ecological adaptations, and expand molecular datasets for a more comprehensive understanding of Funiliomycetaceae.

Species within Funiliomycetaceae exhibit considerable ecological diversity in lifestyle, host range, and geographical distribution. The family includes endophytic, saprobic, and epiphytic species, colonizing hosts from a broad spectrum of plant families such as Fabaceae, Ericaceae, Poaceae, and Rutaceae, as well as decaying unidentified leaves or ornamentals, indicating no strict host specificity. Geographically, Funiliomycetaceae species are widely distributed across multiple continents, including North America, Asia, and South America, and occur in diverse climatic zones ranging from temperate to tropical regions (Table 4). By contrast, most Nothodactylariaceae species are saprobic, with the exception of Nothodactylaria nephrolepidis, which is epiphytic. They show a strong preference for ferns (e.g., Blechnaceae, Lomariopsidaceae) and are known only from China and South Africa, mostly in tropical and subtropical climates (Zhang et al. 2025). These marked differences in host range, ecological strategy, geographical distribution, and climatic preference underscore the distinct ecological niche occupied by Funiliomycetaceae. With the addition of Funiliomycetaceae, the order now comprises 21 families, highlighting both the considerable diversity within Amphisphaeriales and the presence of previously unrecognized lineages.

The new family comprises Funiliomyces biseptatus, ten previously described “Dactylaria” species, and our newly obtained isolates. As confirmed by phylogenetic analyses (Fig. 1), these ten “Dactylaria” species are not congeneric with the type of Dactylaria (D. purpurella) but form a coherent, monophyletic group with F. biseptatus. This supports their recognition as a single genus, consistent with the “One Fungus, One Name” principle (Hawksworth 2012) and the general taxonomic guideline of recognizing monophyletic genera with coherent morphological characters. Accordingly, we transfer the ten previously described “Dactylaria” species into the genus Funiliomyces as new combinations (F. acaciae, F. bisepatus, F. calliandrae, F. fragilis, F. hwasunensis, F. mavisleverae, F. monticola, F. retrophylli, F. sparsus, and F. zapatensis). Furthermore, Funiliomyces is designated as the type genus of Funiliomycetaceae. The genus was described by Aptroot (2004), containing only its type species, F. biseptatus, which was collected from dead leaves of a Bromeliaceae species in a rock field. This species differs from the other newly transferred species in the genus by possessing only a sexual morph. In the present study, we summarize the asexual morphological features, lifestyle, host associations, and distribution of Funiliomyces species (Table 4), providing a comprehensive framework for future identification, ecological studies, and taxonomic research of the genus.

In addition to transferring ten “Dactylaria” species to Funiliomyces, we describe a new species (F. jiangxiensis) that further enriches the diversity of this genus. Multi-gene phylogenetic analyses also indicate that the new isolates, Funiliomyces jiangxiensis, occupies a distinct position within Funiliomyces, supported by high statistical values (SH-aLRT/UFB/PP = 98.8/100/0.90). Morphologically, F. jiangxiensis can be distinguished from its closely related species (F. hwasunensis and F. calliandrae) by differences in conidiophore structure, conidiogenous cell denticulation, and conidial characteristics (Table 4). Additionally, Funiliomyces jiangxiensis was isolated from the medicinal plant Tetradium ruticarpum. This plant, commonly known as Evodia, is renowned for its therapeutic properties, including anti-inflammatory, analgesic, and antimicrobial activities (Xiao et al. 2023). Endophytes have been studied and are known to produce novel biologically active compounds (Liao et al. 2025). For example, Funiliomyces hwasunensis CMML 20-35, isolated from the roots of Zoysia japonica, inhibited the mycelial growth of Rhizoctonia solani AG2-2(IIIB) by 37.97% in dual culture (Liu et al. 2025). To date, endophytic fungal studies on this host are limited, with only a few taxa reported recently, including Cyphellophora guangxiensis and Pseudokeissleriella tetradii (Mi et al. 2025a, 2025b). These findings emphasize the need for comprehensive surveys and further functional studies to explore the diversity and antimicrobial capabilities of endophytic fungi associated with this medicinal plant.

Supplementary Material

XML Treatment for Funiliomycetaceae

XML Treatment for Funiliomyces

XML Treatment for Funiliomyces jiangxiensis

XML Treatment for Funiliomyces acaciae

XML Treatment for Funiliomyces bisepatus

XML Treatment for Funiliomyces calliandrae

XML Treatment for Funiliomyces fragilis

XML Treatment for Funiliomyces hwasunensis

XML Treatment for Funiliomyces mavisleverae

XML Treatment for Funiliomyces monticola

XML Treatment for Funiliomyces retrophylli

XML Treatment for Funiliomyces sparsus

XML Treatment for Funiliomyces zapatensis