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urn:lsid:arphahub.com:pub:C004A564-9D6A-5F9F-B058-6A3815DFE9C3
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MycoKeys
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10.3897/mycokeys.108.128983
128983
Research Article
Ascomycota
Diaporthaceae
Diaporthales
Sordariomycetes
Taxonomy
Asia
﻿New species of Diaporthe (Diaporthaceae, Diaporthales) from Bauhiniavariegata in China
Zhu Yaquan https://orcid.org/0000-0002-3296-239X
1
Ma Lei 2
Xue Han https://orcid.org/0000-0003-0414-6237
1
Li Yong https://orcid.org/0000-0002-4406-1329
1
Jiang Ning n.jiang@caf.ac.cn
https://orcid.org/0000-0002-9656-8500
1
1 Key Laboratory of Biodiversity Conservation of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, China Ecology and Nature Conservation Institute, Chinese Academy of Forestry Beijing China
2 Forest Pest Control and Quarantine Station of Tonghua County, Tonghua 134001, China Forest Pest Control and Quarantine Station of Tonghua County Tonghua China
Corresponding author: Ning Jiang (n.jiang@caf.ac.cn)
Academic editor: Huzefa Raja

2024
13 9 2024
108 317335
79C5782D-30B4-54AD-A854-C2EE13AFB24C04 6 2024
07 8 2024
Yaquan Zhu, Lei Ma, Han Xue, Yong Li, Ning Jiang
https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
﻿Abstract

Diaporthe species are known as endophytes, saprobes and pathogens infecting a wide range of plants and resulting in important crop diseases. In the present study, four strains of Diaporthe were obtained from diseased leaves of Bauhiniavariegata in Guangdong Province, China. Phylogenetic analyses were conducted to identify these strains using five gene regions: internal transcribed spacer (ITS), calmodulin (cal), histone H3 (his3), translation elongation factor 1-α (tef1) and β-tubulin (tub2). The results combined with morphology revealed two new species of Diaporthe named D.bauhiniicola in D.arecae species complex and D.guangzhouensis in D.sojae species complex.

Key words: Diaporthales
morphology
multi-gene phylogeny
taxonomy
two new taxa
This study was supported by Fundamental Research Funds of CAF (CAFYBB2023PA002), and the National Microbial Resource Center of the Ministry of Science and Technology of the People’s Republic of China (NMRC-2023-7).Citation

Zhu Y, Ma L, Xue H, Li Y, Jiang N (2024) New species of Diaporthe (Diaporthaceae, Diaporthales) from Bauhinia variegata in China. MycoKeys 108: 317–335. https://doi.org/10.3897/mycokeys.108.128983
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pmc﻿Introduction

Diaporthe (syn. Phomopsis) is the type genus of Diaporthaceae in Diaporthales (Hyde et al. 2014; Maharachchikumbura et al. 2016). Before the implementation of “one fungus, one name”, it has been a common practice to use two names for the fungal species with pleomorphic life cycles (Taylor 2011). The genus Diaporthe established in 1870 predates Phomopsis established in 1905, thus Diaporthe is recommended for use (Rossman et al. 2015). More than 1200 epithets for Diaporthe have been listed in Index Fungorum with names often based on host association (http://www.indexfungorum.org/, accessed June 2024).

The teleomorph of Diaporthe is characterized by aggregated spherical ascomata with tapering necks, unitunicate, 8-spored, elongate to clavate asci, and septate or aseptate, elongated to elliptical, hyaline ascospores with larger guttules at center and smaller ones at the ends (Senanayake et al. 2018; Yang et al. 2020). The anamorph is characterized by black, ostiolate pycnidia containing cylindrical phialides often producing three types of hyaline, aseptate conidia called α-conidia, β-conidia and γ-conidia (Udayanga et al. 2012a; Dissanayake et al. 2017; Fan et al. 2018). The α-conidia and β-conidia are produced frequently, but the γ-conidia are rarely observed (Gomes et al. 2013).

Diaporthe species are associated with a wide range of plant hosts as pathogens, endophytes and saprobes of crops, forest trees and ornamentals (Farr et al. 2002a; Crous 2005; Udayanga et al. 2012b, 2014a, 2014b, 2015; Jiang et al. 2021; Zhu et al. 2023). As plant pathogens, Diaporthe species cause severe diseases, e.g., leaf spots, blights, dieback, scab, decay, stem end rots and wilt of many economically important plants including species of Citrus (Guarnaccia and Crous 2017), Macadamia (Wrona et al. 2020), Rosa (Caio et al. 2021), Vaccinium (Farr et al. 2002b), Vitis (Manawasinghe et al. 2019) and many more (Yang et al. 2018, 2021; Guarnaccia et al. 2020; Guo et al. 2020; Ariyawansa et al. 2021). In addition, Diaporthe species can live inside the healthy host tissues as endophytes (Huang et al. 2015; Dong et al. 2021). In addition, species of Diaporthe have been also reported as saprobes from different woody hosts (Dissanayake et al. 2020).

Species identification of Diaporthe has traditionally been based on host as well as morphological characters such as the size and shape of fruiting bodies and spores (Mostert et al. 2001; Santos and Phillips 2009). However, recent studies have shown that many species of Diaporthe are not host-specific i.e., one species may infect more than one host species (Vrandecic et al. 2011; Bai et al. 2015; Zhang et al. 2018; Huang et al. 2021; Sun et al. 2021; Cao et al. 2022). Moreover, many Diaporthe species that are morphologically similar have proven to be genetically distinct (van Rensburg et al. 2006; Yang et al. 2018). Phylogenetic analysis using a five-locus dataset (ITS-tef1-tub2-cal-his3) has been widely used to identify species of Diaporthe species (Santos et al. 2017; Marin-Felix et al. 2019; Hilário et al. 2021b; Norphanphoun et al. 2022). Diaporthe was clustered into 13 groups, namely D.arecae, D.biconispora, D.carpini, D.decedens, D.eres, D.oncostoma, D.pustulata, D.rudis, D.scobina, D.sojae, D.toxica, D.varians and D.vawdreyi species complexes and nine singletons as D.acerina, D.acutispora, D.crataegi, D.multiguttulata, D.ocoteae, D.perjuncta, D.pseudoalnea, D.spartinicola and D.undulata based on multilocus phylogeny (Norphanphoun et al. 2022; Hongsanan et al. 2023).

Bauhiniavariegata is a flowering plant species belonging to Fabaceae. It is native to China and cultivated as an ornamental tree in subtropical and tropical climate for its scented flowers. The aim of the present study was to identify new isolates collected from diseased leaves of Bauhiniavariegata in China following the combined approaches of morphology and phylogeny in the genus Diaporthe.

﻿Materials and methods

﻿Isolation and morphological characterization

In 2022, a plant disease investigation was conducted in Guangdong Province, China. Small and irregular leaf spots were observed on the leaves of Bauhiniavariegata, and 14 leaves were collected for isolation. The leaves were firstly surface-sterilized for 1 min in 75% ethanol, 3 min in 1.25% sodium hypochlorite and 1 min in 75% ethanol, rinsed for 2 min in distilled water and blotted on dry sterile filter paper. Then, the discolored areas were cut into 0.5 × 0.5 cm pieces and transferred to the surface of potato dextrose agar plates (PDA; 200 g potatoes, 20 g dextrose, 20 g agar per litre), incubated at 25 °C to obtain pure cultures. The cultures were deposited in the China Forestry Culture Collection Center (CFCC; http://cfcc.caf.ac.cn/) and the specimen was deposited in the Herbarium of the Chinese Academy of Forestry (CAF; http://museum.caf.ac.cn/).

The isolates were grown on PDA, MEA and SNA plates, incubated at 25 °C under a 12 h near-ultraviolet light/12 h dark cycle to induce sporulation. Colony characters and pigment production on PDA, MEA and SNA were noted for the 10-day culture. Microscopic structures of the fungi growing on medium were mounted in water and examined under an Axio Imager 2 microscope (Zeiss, Oberkochen, Germany). At least 30 measurements were made for each structure examined.

﻿DNA extraction, amplification and sequencing

The genomic DNA was extracted from the fresh mycelium harvested from PDA plates after seven days using a cetyltrimethylammonium bromide (CTAB) method (Doyle and Doyle 1990). For initial genus confirmation, the internal transcribed spacer (ITS) region was sequenced. After confirmation of Diaporthe species, four additional gene regions coding for translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), calmodulin (cal) and his-tone H3 (his3) were sequenced. The primer pairs and amplification conditions for each of the above-mentioned gene regions are provided in Table 1.

Table 1. Loci assessed in this study with used PCR primers and program.

Loci	Primers	PCR: Thermal Cycles: (Annealing Temp. in Bold)	Reference	
ITS	ITS1f/ITS4	(95 °C: 30 s, 48 °C: 30 s, 72 °C: 1 min) × 35 cycles	White et al. 1990	
cal	CAL228F/CAL737R	(95 °C: 15 s, 54 °C: 20 s, 72 °C: 1 min) × 35 cycles	Carbone and Kohn 1999	
his3	CYLH3F/H3-1b	(95 °C: 30 s, 57 °C: 30 s, 72 °C: 1 min) × 35 cycles	Crous et al. 2004; Glass and Donaldson 1995	
tef1	EF1-728F/EF1-986R	(95 °C: 15 s, 54 °C: 20 s, 72 °C: 1 min) × 35 cycles	Carbone and Kohn 1999	
tub2	T1(Bt2a)/Bt2b	(95 °C: 30 s, 55 °C: 30 s, 72 °C: 1 min) × 35 cycles	Glass and Donaldson 1995; O’Donnell and Cigelnik 1997	

A PCR reaction was conducted in a 20 µL reaction volume, and the components were as follows: 1 µL DNA template (20 ng/μL), 1 µL forward 10 µM primer, 1 µL reverse 10 µM primer, 10 µL T5 Super PCR Mix (containing Taq polymerase, dNTP and Mg2+, Beijing Tisingke Biotech Co., Ltd., Beijing, China), and 7 µL sterile water. Amplifications were performed using a T100 Thermal Cycler (Bio-Rad, Hercules, CA, USA). All amplified PCR products were evaluated visually with 1.4% agarose gels stained with ethidium bromide and PCR positive products sent to Sangon Biotech (Shanghai) Co., Ltd., (Beijing, China) for sequencing. Strands were sequenced in both directions using PCR primers. The new sequences generated in this study, as well as the reference sequences of all isolates used in the present study, are listed in Table 2.

Table 2. Isolates and GenBank accession numbers used in the phylogenetic analyses of Diaporthe.

Species	Location	Host	Strain	GenBank Accession Number	
ITS	tef1	tub2	cal	his3	
Diaportheabsenteum	China	Camelliasinensis	LC3429*	KP267897	KP267971	KP293477	NA	KP293547	
D.absenteum	China	Camelliasinensis	LC3564	KP267912	KP267986	KP293492	NA	KP293559	
D.acaciarum	Tanzania	Acaciatortilis	CBS 138862*	KP004460	NA	KP004509	NA	KP004504	
D.acericola	Italy	Acernegundo	MFLUCC 17-0956*	KY964224	KY964180	KY964074	KY964137	NA	
D.aceris	Japan	Acer sp.	LC8112	KY491547	KY491557	KY491567	KY491575	NA	
D.actinidiae	New Zealand	Actinidiadeliciosa	ICMP 13683*	KC145886	KC145941	NA	NA	NA	
D.acuta	China	Pyruspyrifolia	CGMCC 3.19600*	MK626957	MK654802	MK691225	MK691124	MK726161	
D.alangii	China	Alangiumkurzii	CFCC 52556*	MH121491	MH121533	MH121573	MH121415	MH121451	
D.alangii	China	Alangiumkurzii	CFCC 52557	MH121492	MH121534	MH121574	MH121416	MH121452	
D.alnea	Netherlands	Alnus sp.	CBS 146.46	KC343008	KC343734	KC343976	KC343250	KC343492	
D.amaranthophila	Japan	Amaranthustricolor	MAFF 246900	LC459575	LC459577	LC459579	LC459583	LC459581	
D.ambigua	South Africa	Pyruscommunis	CBS 114015*	KC343010	KC343736	KC343978	KC343252	KC343494	
D.angelicae	Austria	Heracleumsphondylium	CBS 111592*	KC343027	KC343753	KC343995	KC343269	KC343511	
D.anhuiensis	China	Cunninghamialanceolata	CNUCC 201901*	MN219718	MN224668	MN227008	MN224549	MN224556	
D.arctii	Austria	Arctiumlappa	CBS 139280*	KJ590736	KJ590776	KJ610891	KJ612133	KJ659218	
D.arecae	India	Arecacatechu	CBS 161.64*	KC343032	KC343758	KC344000	KC343274	KC343516	
D.arengae	Hong Kong	Arengaengleri	CBS 114979*	KC343034	KC343760	KC344002	KC343276	KC343518	
D.arezzoensis	Italy	Cytisus sp.	MFLUCC 15-0127	MT185503	NA	NA	NA	NA	
D.aseana	Thailand	Unidentified dead leaf	MFLUCC 12-0299a*	KT459414	KT459448	KT459432	KT459464	NA	
D.australiana	Australia	Macadamia	CBS 146457	MN708222	MN696522	MN696530	NA	NA	
D.bauhiniicola	China	Bauhiniavariegata	CFCC 58154*	PP864723	PP938599	PP938603	PP938607	PP938611	
D.bauhiniicola	China	Bauhiniavariegata	GZ13B	PP864724	PP938600	PP938604	PP938608	PP938612	
D.batatas	USA	Ipomoeabatatas	CBS 122.21*	KC343040	KC343766	KC344008	KC343282	KC343524	
D.beilharziae	Australia	Indigoferaaustralis	BRIP 54792*	JX862529	JX862535	KF170921	NA	NA	
D.biconispora	China	Citrusgrandis	ZJUD62	KJ490597	KJ490476	KJ490418	MT227578	KJ490539	
D.biguttulata	China	Citruslimon	ZJUD47*	KJ490582	KJ490461	KJ490403	NA	KJ490524	
D.brasiliensis	Brazil	Aspidosperma sp.	CBS 133183*	KC343042	KC343768	KC344010	KC343284	KC343526	
D.caatingaensis	Brazil	Tacingainamoena	CBS 141542*	KY085927	KY115603	KY115600	NA	KY115605	
D.camelliae-oleiferae	China	Camelliaoleifera	HNZZ027*	MZ509555	MZ504707	MZ504718	MZ504685	MZ504696	
D.caryae	China	Caryaillinoensis	CFCC 52563*	MH121498	MH121540	MH121580	MH121422	MH121458	
D.caryae	China	Caryaillinoensis	CFCC 52564	MH121499	MH121541	MH121581	MH121423	MH121459	
D.cercidis	China	Cercischinensis	CFCC 52565*	MH121500	MH121542	MH121582	MH121424	MH121460	
D.cercidis	China	Cercischinensis	CFCC 52566	MH121501	MH121543	MH121583	MH121425	MH121461	
D.chiangraiensis	Thailand	Bauhinia sp.	MFLUCC 17-1669*	MF190119	MF377598	NA	NA	NA	
D.chrysalidocarpi	China	Chrysalidocarpuslutescens	SAUCC194.35	MT822563	MT855760	MT855876	MT855646	MT855532	
D.cichorii	Italy	Cichoriumintybus	MFLUCC 17-1023*	KY964220	KY964176	KY964104	KY964133	NA	
D.cinmomi	China	Cinnamomum sp.	CFCC 52569*	MH121504	MH121546	MH121586	NA	MH121464	
D.cinmomi	China	Cinnamomum sp.	CFCC 52570	MH121505	MH121547	MH121587	NA	MH121465	
D.citriasiana	China	Citrusunshiu	CGMCC 3.15224*	JQ954645	JQ954663	KC357459	KC357491	KJ490515	
D.columnaris	USA	Vacciniumvitisidaea	AR3612*	AF439625	NA	NA	NA	NA	
D.compacta	China	Camelliasinensis	CGMCC 3.17536*	KP267854	KP267928	KP293434	NA	KP293508	
D.convolvuli	Turkey	Convolvulusarvensis	CBS 124654*	KC343054	KC343780	KC344022	KC343296	KC343538	
D.cucurbitae	Canada	Cucumis sp.	DAOM 42078*	KM453210	KM453211	KP118848	NA	KM453212	
D.cuppatea	South Africa	Aspalathuslinearis	CBS 117499*	KC343057	KC343783	KC344025	KC343299	KC343541	
D.cyatheae	Taiwan	Cyathealepifera	YMJ 1364*	JX570889	KC465406	KC465403	KC465410	NA	
D.discoidispora	China	Citrusunshiu	ZJUD89*	KJ490624	KJ490503	KJ490445	NA	KJ490566	
D.drenthii	Australia	Macadamia	CBS 146453	MN708229	MN696526	MN696537	NA	NA	
D.durionigena	Vietnam	Duriozibethinus	VTCC 930005	MN453530	MT276157	MT276159	NA	NA	
D.endocitricola	China	Citrusmaxima	ZHKUCC20-0012*	MT355682	MT409336	MT409290	MT409312	NA	
D.endophytica	Brazil	Schinusterebinthifolius	CBS 133811*	KC343065	KC343791	KC344033	KC343307	KC343549	
D.eucalyptorum	China	Eucalyptus	CBS 132525*	MH305525	NA	NA	NA	NA	
D.eugeniae	Indonesia	Eugeniaaromatica	CBS 444.82*	KC343098	KC343824	KC344066	KC343340	KC343582	
D.fraxini-angustifoliae	Australia	Fraxinusangustifolia	BRIP 54781*	JX862528	JX862534	KF170920	NA	NA	
D.fructicola	Japan	Passifloraedulis × P. edulis	MAFF 246408*	LC342734	LC342735	LC342736	LC342738	LC342737	
D.fulvicolor	China	Pyruspyrifolia	CGMCC 3.19601*	MK626859	MK654806	MK691236	MK691132	MK726163	
D.ganjae	USA	Cannabissativa	CBS 180.91*	KC343112	KC343838	KC344080	KC343354	KC343596	
D.goulteri	Australia	Helianthusannuus	BRIP 55657a*	KJ197290	KJ197252	KJ197270	NA	NA	
D.guangdongensis	China	Citrusmaxima	ZHKUCC20-0014*	MT355684	MT409338	MT409292	MT409314	NA	
D.guangxiensis	China	Vitisvinifera	JZB320094*	MK335772	MK523566	MK500168	MK736727	NA	
D.guangzhouensis	China	Bauhiniavariegataa	CFCC 58151*	PP864725	PP938601	PP938605	PP938609	PP938613	
D.guangzhouensis	China	Bauhiniavariegata	GZ13E	PP864726	PP938602	PP938606	PP938610	PP938614	
D.gulyae	Australia	Helianthusannuus	BRIP 54025*	JF431299	JN645803	KJ197271	NA	NA	
D.guttulata	China	Unknown	CGMCC 3.20100	MT385950	MT424685	MT424705	MW022470	MW022491	
D.helianthi	Serbia	Helianthusannuus	CBS 592.81*	KC343115	KC343841	KC344083	KC343357	KC343599	
D.heterostemmatis	China	Heterostemmagrandiflorum	SAUCC194.85*	MT822613	MT855925	MT855810	MT855692	MT855581	
D.hongkongensis	China	Dichroafebrífuga	CBS 115448*	KC343119	KC343845	KC344087	KC343361	KC343603	
D.hordei	Norway	Hordeumvulgare	CBS 481.92*	KC343120	KC343846	KC344088	KC343362	KC343604	
D.huangshanensis	China	Camelliaoleifera	CNUCC 201903*	MN219729	MN224670	MN227010	NA	MN224558	
D.hubeiensis	China	Vitisvinifera	JZB320123	MK335809	MK523570	MK500148	MK500235	NA	
D.hunanensis	China	Camelliaoleifera	HNZZ023*	MZ509550	MZ504702	MZ504713	MZ504680	MZ504691	
D.infecunda	Brazil	Schinus sp.	CBS 133812*	KC343126	KC343852	KC344094	KC343368	KC343610	
D.infertilis	Suriname	Camelliasinensis	CBS 230.52*	KC343052	KC343778	KC344020	KC343294	KC343536	
D.kochmanii	Australia	Helianthusannuus	BRIP 54033*	JF431295	JN645809	NA	NA	NA	
D.kongii	Australia	Portulacagrandifla	BRIP 54031*	JF431301	JN645797	KJ197272	NA	NA	
D.krabiensis	Thailand	marine based habitats	MFLUCC 17-2481*	MN047101	MN433215	MN431495	NA	NA	
D.leucospermi	Australia	Leucospermum sp.	CBS 111980*	JN712460	KY435632	KY435673	KY435663	KY435653	
D.limonicola	Malta	Citruslimon	CPC 28200*	NR_154980	MF418501	MF418582	MF418256	MF418342	
D.litchiicola	Australia	Litchichinensis	BRIP 54900*	JX862533	JX862539	KF170925	NA	NA	
D.lithocarpi	China	Lithocarpusglabra	CGMCC 3.15175*	KC153104	KC153095	KF576311	KF576235	NA	
D.longicolla	USA	Glycinemax	FAU599*	KJ590728	KJ590767	KJ610883	KJ612124	KJ659188	
D.longispora	Canada	Ribes sp.	CBS 194.36*	KC343135	KC343861	KC344103	KC343377	KC343619	
D.lusitanicae	Portugal	Foeniculumvulgare	CBS 123212	KC343136	KC343862	KC344104	KC343378	KC343620	
D.lusitanicae	Portugal	Foeniculumvulgare	CBS 123213*	MH863280	KC343863	KC344105	KC343379	KC343621	
D.malorum	Portugal	Malusdomestica	CAA 734*	KY435638	KY435627	KY435668	KY435658	KY435648	
D.manihotia	Rwanda	Manihotutilissima	CBS 505.76	KC343138	KC343864	KC344106	KC343380	KC343622	
D.masirevicii	Australia	Helianthusannuus	BRIP 57892a*	KJ197276	KJ197239	KJ197257	NA	NA	
D.mayteni	Brazil	Maytenusilicifolia	CBS 133185	KC343139	KC343865	KC344107	KC343381	KC343623	
D.megalospora	Not stated	Sambucuscanadensis	CBS 143.27	KC343140	KC343866	KC344108	KC343382	KC343624	
D.melitensis	Malta	Citruslimon	CPC 27873*	MF418424	MF418503	MF418584	MF418258	MF418344	
D.melonis	USA	Cucumismelo	CBS 507.78*	KC343142	KC343868	KC344110	KC343384	KC343626	
D.melonis	Indonesia	Glycinesoja	CBS 435.87	KC343141	KC343867	KC344109	KC343383	KC343625	
D.middletonii	Australia	Rapistrumrugostrum	BRIP 54884e*	KJ197286	KJ197248	KJ197266	NA	NA	
D.millettiae	China	Millettiareticulata	GUCC9167*	MK398674	MK480609	MK502089	MK502086	NA	
D.minusculata	China	saprobic on decaying wood	CGMCC 3.20098*	MT385957	MT424692	MT424712	MW022475	MW022499	
D.miriciae	Australia	Helianthusannuus	BRIP 54736j*	KJ197282	KJ197244	KJ197262	NA	NA	
D.musigena	Australia	Musa sp.	CBS 129519*	KC343143	KC343869	KC344111	KC343385	KC343267	
D.myracrodruonis	Brazil	Astroniumurundeuva	URM 7972*	MK205289	MK213408	MK205291	MK205290	17	
D.nelumbonis	Taiwan	Nelumbonucifera	R. Kirschner 4114*	KT821501	NA	LC086652	NA	NA	
D.neoarctii	USA	Ambrosiatrifi	CBS 109490*	KC343145	KC343871	KC344113	KC343387	KC343629	
D.neoraonikayaporum	Thailand	Tectonagrandis	MFLUCC 14-1136*	KU712449	KU749369	KU743988	KU749356	NA	
D.oculi	Japan	Homosapiens	HHUF 30565*	LC373514	LC373516	LC373518	NA	NA	
D.osmanthi	China	Osmanthusfragrans	GUCC9165*	MK398675	MK480610	MK502091	MK502087	NA	
D.ovalispora	China	Citruslimon	CGMCC 3.17256*	KJ490628	KJ490507	KJ490449	NA	KJ490570	
D.oxe	Brazil	Maytenusilicifolia	CBS 133186*	KC343164	KC343890	KC344132	KC343406	KC343648	
D.pandanicola	Thailand	Pandanus sp.	MFLUCC 17-0607*	MG646974	NA	MG646930	NA	NA	
D.paranensis	Brazil	Maytenusilicifolia	CBS 133184*	KC343171	KC343897	KC344139	KC343413	KC343655	
D.pascoei	Australia	Perseaamericana	BRIP 54847*	JX862532	JX862538	KF170924	NA	NA	
D.passiflorae	South America	Passiflaedulis	CBS 132527*	JX069860	KY435633	KY435674	KY435664	KY435654	
D.passifloricola	Malaysia	Passiflorafoetida	CBS 141329*	KX228292	NA	KX228387	NA	KX228367	
D.perseae	Netherlands	Perseagratissima	CBS 151.73*	KC343173	KC343899	KC343141	KC343415	KC343657	
D.pescicola	China	Prunuspersica	MFLUCC 16-0105*	KU557555	KU557623	KU557579	KU557603	NA	
D.phaseolorum	USA	Phaseolusvulgaris	AR4203*	KJ590738	KJ590739	KJ610893	KJ612135	KJ659220	
D.phoenicicola	India	Arecacatechu	CBS 161.64*	MH858400	GQ250349	JX275440	JX197432	NA	
D.podocarpi-macrophylli	China	Podocarpusmacrophyllus	CGMCC 3.18281*	KX986774	KX999167	KX999207	KX999278	KX999246	
D.pseudobauhiniae	Thailand	Bauhinia sp.	MFLU 17-1670	MF190118	MF377599	NA	NA	NA	
D.pseudobauhiniae	Thailand	Bauhinia sp.	MFLUCC 17-1669*	MF190119	MF377598	NA	NA	NA	
D.pseudolongicolla	Serbia	Glycinemax	PL42*	JQ697843	JQ697856	NA	NA	NA	
D.pseudolongicolla	Croatia	Glycinemax	CBS 127269	KC343155	KC343881	KC344123	KC343397	KC343639	
D.pseudomangiferae	Dominican Republic	Mangiferaindica	CBS 101339*	KC343181	KC343907	KC344149	KC343423	KC343665	
D.pseudooculi	Japan	Homosapiens	HHUF 30617*	NR_161019	LC373517	LC373519	NA	NA	
D.pseudophoenicicola	Spain	Phoenixdactylifera	CBS 462.69*	KC343184	KC343910	KC344152	KC343426	KC343668	
D.pseudophoenicicola	Iraq	Mangiferaindica	CBS 176.77	KC343183	KC343909	KC344151	KC343425	KC343667	
D.pterocarpicola	Thailand	Pterocarpusindicus	MFLUCC 10-0580a*	JQ619887	JX275403	JX275441	JX197433	NA	
D.pyracanthae	Portugal	Pyracanthacoccinea	CBS 142384*	KY435635	KY435625	KY435666	KY435656	KY435646	
D.racemosae	South Africa	Euclearacemosa	CPC 26646*	MG600223	MG600225	MG600227	MG600219	MG600221	
D.raonikayaporum	Brazil	Spondiasmombin	CBS 133182*	KC343188	KC343914	KC344156	KC343430	KC343672	
D.rhodomyrti	China	Rhodomyrtustomentosa	CFCC 53101	MK432643	MK578119	MK578046	MK442965	MK442990	
D.rhodomyrti	China	Rhodomyrtustomentosa	CFCC 53102	MK432644	MK578120	MK578047	MK442966	MK442991	
D.rosae	Thailand	Rosa sp.	MFLUCC 17-2658*	MG828894	NA	MG843878	MG829273	NA	
D.rosiphthora	Brazil	Rosa sp.	COAD 2914*	MT311197	MT313693	NA	MT313691	NA	
D.rossmaniae	Portugal	Vacciniumcorymbosum	CAA762*	MK792290	MK828063	MK837914	MK883822	MK871432	
D.sackstonii	Australia	Helianthusannuus	BRIP 54669b*	KJ197287	KJ197249	KJ197267	NA	NA	
D.salinicola	Thailand	Xylocarpus sp.	MFLU 18-0553*	MN047098	MN077073	NA	NA	NA	
D.sambucusii	China	Sambucuswilliamsii	CFCC 51986*	KY852495	KY852507	KY852511	KY852499	KY852503	
D.sambucusii	China	Sambucuswilliamsii	CFCC 51987	KY852496	KY852508	KY852512	KY852500	KY852504	
D.schimae	China	Schimasuperba	CFCC 53103*	MK432640	MK578116	MK578043	MK442962	MK442987	
D.schimae	China	Schimasuperba	CFCC 53104	MK432641	MK578117	MK578044	MK442963	MK442988	
D.schini	Brazil	Schinusterebinthifolius	CBS 133181*	KC343191	KC343917	KC344159	KC343433	KC343675	
D.schoeni	Italy	Schoenusnigricans	MFLU 15-1279*	KY964226	KY964182	KY964109	KY964139		
D.sclerotioides	Netherlands	Cucumissativus	CBS 296.67*	KC343193	KC343919	KC344161	KC343435	KC343677	
D.searlei	Australia	Macadamia	CBS 146456*	MN708231	NA	MN696540	NA	NA	
D.sennae	China	Sennabicapsularis	CFCC 51636*	KY203724	KY228885	KY228891	KY228875	NA	
D.sennae	China	Sennabicapsularis	CFCC 51637	KY203725	KY228886	KY228892	KY228876	NA	
D.serafiniae	Australia	Helianthusannuus	BRIP 55665a*	KJ197274	KJ197236	KJ197254	NA	NA	
D.siamensis	Thailand	Dasymaschalon sp.	MFLUCC 10-0573a*	JQ619879	JX275393	JX275429	JX197423	NA	
D.sinensis	China	Amaranthus sp.	ZJUP0033-4*	MK637451	MK660449	MK660447	NA	MK660451	
D.sojae	USA	Glycinemax	FAU635*	KJ590719	KJ590762	KJ610875	KJ612116	KJ659208	
D.spinosa	China	Pyruspyrifolia	CGMCC 3.19602*	MK626849	MK654811	MK691234	MK691129	MK726156	
D.stewartii	Not stated	Cosmosbipinnatus	CBS 193.36*	MH867279	GQ250324	JX275421	JX197415	NA	
D.subellipicola	China	On dead wood	KUMCC 17-0153*	MG746632	MG746633	MG746634	NA	NA	
D.subordinaria	New Zealand	Plantagolanceolata	CBS 464.90*	KC343214	KC343940	KC344182	KC343456	KC343698	
D.taiwanensis	Taiwan	Ixorachinensis	NTUCC 18-105-1*	MT241257	MT251199	MT251202	MT251196	NA	
D.taoicola	China	Prunuspersica	MFLUCC 16-0117*	KU557567	KU557635	KU557591	NA	NA	
D.tarchonanthi	South Africa	Tarchonanthuslittoralis	CBS 146073*	MT223794	NA	MT223733	NA	MT223759	
D.tecomae	Brazil	Tabebuia sp.	CBS 100547*	KC343215	KC343941	KC344183	KC343457	KC343699	
D.tectonae	Thailand	Tectonagrandis	MFLUCC 12-0777*	KU712430	KU749359	KU743977	KU749345	NA	
D.tectonendophytica	Thailand	Tectonagrandis	MFLUCC 13-0471*	KU712439	KU749367	KU743986	KU749354	NA	
D.tectonigena	China	Tectonagrandis	MFLUCC 12-0767*	KU712429	KU749371	KU743976	KU749358	NA	
D.tectonigena	China	Camelliasinensis	LC6512	KX986782	KX999174	KX999214	KX999284	KX999254	
D.terebinthifolii	Brazil	Schinusterebinthifolius	CBS 133180*	KC343216	KC343942	KC344184	KC343458	KC343700	
D.thunbergiicola	Thailand	Thunbergialaurifolia	MFLUCC 12-0033*	KP715097	KP715098	NA	NA	NA	
D.tulliensis	Australia	Theobromacacao	BRIP 62248a*	KR936130	KR936133	KR936132	NA	NA	
D.ueckeri	USA	Cucumismelo	FAU656*	KJ590726	KJ590747	KJ610881	KJ612122	KJ659215	
D.unshiuensis	China	Fortunellamargarita	CGMCC 3.17566*	KJ490584	KJ490463	KJ490405	NA	KJ490526	
D.unshiuensis	China	Caryaillinoensis	CFCC 52594	MH121529	MH121571	MH121606	MH121447	MH121487	
D.unshiuensis	China	Caryaillinoensis	CFCC 52595	MH121530	MH121572	MH121607	MH121448	MH121488	
D.vawdreyi	Australia	Psidiumguajava	BRIP 57887a	KR936126	KR936129	KR936128	NA	NA	
D.vexans	USA	Solanummelongena	CBS 127.14	KC343229	KC343955	KC344197	KC343471	KC343713	
D.viniferae	China	Vitisvinifera	JZB320071*	MK341550	MK500107	MK500112	MK500119	NA	
D.vochysiae	Brazil	Vochysiadivergens	LGMF1583*	MG976391	MK007526	MK007527	MK007528	MK033323	
D.xishuangbanica	China	Camelliasinensis	CGMCC 3.18283*	KX986784	KX999176	KX999217	NA	NA	
D.xishuangbanica	China	Camelliasinensis	LC6707	KX986783	KX999175	KX999216	NA	KX999255	
Notes: NA, not applicable. * ex-type strains.

﻿Phylogeny

For the phylogenetic analysis, sequences of reference Diaporthe species and related taxa were downloaded from NCBI GenBank based on recent publications on the genus Diaporthe (Norphanphoun et al. 2022) (Table 2). Downloaded sequences were aligned together with the sequences obtained in the present study using MAFFT version 7.526 (Katoh and Standley 2013) and manually corrected using Bioedit 7.0.9.0 (Hall 1999). The phylogenetic analyses of the combined gene regions were performed using Maximum Likelihood (ML) and Bayesian Inference (BI) methods. ML was conducted using PhyML v. 3.0 (Guindon et al. 2010), with 1000 bootstrap replicates while BI was performed using a Markov Chain Monte Carlo (MCMC) algorithm in MrBayes v. 3.0 (Ronquist and Huelsenbeck 2003). Two MCMC chains, started from random trees for 1,000,000 generations and trees, were sampled every 100th generation, resulting in a total of 10,000 trees. The first 25% of trees were discarded as burn-in of each analysis. Branches with significant Bayesian Posterior Probabilities (BPP) were estimated in the remaining 7500 trees. Phylogenetic trees were visualized with FigTree v.1.3.1 (Rambaut and Drummond 2010) and processed by Adobe Illustrator CS5. The nucleotide sequence data of the new taxa were deposited in GenBank (Table 2)

﻿Results

﻿Phylogenetic analyses

In the present study, we inferred a genus tree of Diaporthe covering a large proportion of sequence data available as last summarized by Norphanphoun et al. (2022). Two strains CFCC 58154 and GZ13B formed a clade in the D.arecae species complex, and the other strains CFCC 58151 and GZ13E in the D.sojae species complex.

In the D.arecae species complex, the combined sequence alignments comprised 61 strains, with D.eucalyptorum (CBS 13252), D.biconispora (ZJUD62) and D.vawdreyi (BRIP 57887a) as the outgroup taxa. The dataset comprised 2662 characters including alignment gaps (590 for ITS, 499 for cal, 485 for his3, 375 for tef1 and 713 for tub2). CFCC 58154 and GZ13B from Bauhiniavariegata formed a distinct clade close to D.sennae (Fig. 1). In the D.sojae species complex, the combined sequence alignments comprised 166 strains (Fig. 2), with D.aceris (LC8112) and D.alnea (CBS 146.46) as the outgroup taxa. The dataset comprised 3025 characters including alignment gaps (602 for ITS, 592 for cal, 521 for his3, 483 for tef1 and 827 for tub2). CFCC 58151 and GZ13E from B.variegata clustered in a distinct clade close to D.tulliensis (Fig. 2).

10.3897/mycokeys.108.128983.figure1 7824435C-8A72-5D06-9C2A-78BEC4E2481F Figure 1. Phylogram of Diaporthearecae species complex resulting from a maximum likelihood analysis based on a combined matrix of ITS, cal, his3, tef1 and tub2 loci. Numbers above the branches indicate ML bootstrap values (left, ML BS ≥ 50%) and Bayesian posterior probabilities (right, BPP ≥ 0.9). Isolates from the present study are in bold and ex-type strains are marked with *.

https://binary.pensoft.net/fig/1132377

10.3897/mycokeys.108.128983.figure2 A1B7D625-34C4-5048-AF00-CACCBFAFB06A Figure 2. Phylogram of Diaporthesojae species complex resulting from a maximum likelihood analysis based on a combined matrix of ITS, cal, his3, tef1 and tub2 loci. Numbers above the branches indicate ML bootstrap values (left, ML BS ≥ 50%) and Bayesian posterior probabilities (right, BPP ≥ 0.9). Isolates from the present study are in bold and ex-type strains are marked with *.

https://binary.pensoft.net/fig/1132378

﻿Taxonomy

Taxon classification Fungi
Diaporthales
Diaporthaceae
﻿ Diaporthe bauhiniicola

Ning Jiang & Y.Q. Zhu sp. nov.

2D6AD0F1-0758-54ED-B0EB-B5BDDB93E656

854183

Fig. 3

Holotype.

China • Guangdong Province, Guangzhou City, Luhu Park, 23°9'11.15"N, 113°16'46.01"E, 92 m asl, on diseased leaves of Bauhiniavariegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, (holotype: CAF800094; ex-type culture: CFCC 58154).

10.3897/mycokeys.108.128983.figure3 2FCBC613-A59C-52FA-9870-2332570318A1 Figure 3. Morphology of DiaporthebauhiniicolaA colonies on PDA, MEA and SNA at 25 °C after 2 weeks B a diseased leaf of BauhiniavariegataC conidioma formed on PDA after 30 days D conidiogenous cells with attached alpha conidia E–G alpha conidia. Scale bars: 200 µm (C); 10 µm (D–J).

https://binary.pensoft.net/fig/1132379

Etymology.

Named after the host genus, Bauhinia.

Description.

Conidiomata formed on PDA pycnidial, scattered to aggregated, black, erumpent, raising above surface of culture medium, subglobose, 150–300 μm diam., exuding white or yellowish creamy conidial droplets from central ostioles after 30 days at 25 °C. Conidiophores reduced to conidiogenous cells. Conidiogenous cells hyaline, unbranched, septate, straight, slightly tapering towards the apex, 6.0–15.0 × 1.5–4.0 μm. Alpha conidia hyaline, aseptate, ellipsoidal to spindle-shaped, biguttulate or with one guttulate, 4.5–7.0 × 2.0–3.0 μm. Beta conidia and gamma conidia not observed. Teleomorph not observed.

Culture characteristics.

Colonies covering entire plate after 2 weeks. On PDA with profuse aerial mycelium, white surface, reverse fulvous. On MEA with fluffy aerial mycelium, dirty white surface, reverse ochreous. On SNA white sparse aerial mycelium, surface and reverse white.

Additional material examined.

China • Guangdong Province, Guangzhou City, Luhu Park, 23°9'11.15"N, 113°16'46.01"E, 92 m asl, on diseased leaves of Bauhiniavariegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, living culture GZ13B.

Notes.

Two strains representing Diaporthebauhiniicola clustered in a clade distinct from its closest phylogenetic neighbour, D.sennae (Fig. 1). D.sennae has been reported from the host Sennabicapsularis in China (Yang et al. 2017). D.bauhiniicola differs from D.sennae by wider alpha conidia (4.5–7.0 × 2.0–3.0 μm in D.bauhiniicola vs. 5.0–6.5 × 1.5–1.8 μm in D.sennae) (Yang et al. 2017). Diaporthebauhiniicola differs from D.sennae in nucleotide sequence data (18/529 in ITS, 5/490 in cal, 15/351 in tef1, 14/677 in tub2) (Yang et al. 2017).

Taxon classification Fungi
Diaporthales
Diaporthaceae
﻿ Diaporthe guangzhouensis

Ning Jiang & Y.Q. Zhu sp. nov.

2A0959D8-088A-5FA5-A7A2-765C956730CF

854184

Fig. 4

Etymology.

Named after the collection site of the type specimen, Guangzhou City.

Holotype.

China • Guangdong Province, Guangzhou City, Longdong straight street, 23°11'41.02"N, 113°22'8.33"E, 46 m asl, on diseased leaves of Bauhiniavariegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, (holotype: CAF800095; ex-type culture: CFCC 58151).

10.3897/mycokeys.108.128983.figure4 B5E70CDE-ED0B-5747-A05F-91B3EFFA11D1 Figure 4. Morphology of DiaportheguangzhouensisA colonies on PDA, MEA and SNA at 25 °C after 2 weeks B the leaf of BauhiniavariegataC conidiomata D conidiogenous cells with attached beta conidia E–G beta conidia. Scale bars: 200 µm (C); 10 µm (D–J).

https://binary.pensoft.net/fig/1132380

Description.

Conidiomata pycnidial, scattered to aggregated, black, erumpent, raising above surface of culture medium, subglobose, 150–450 µm diam, exuding white or yellowish creamy conidial droplets from central ostioles after 30 days at 25 °C. Conidiophores 12.5–24.5 × 1–2.5 μm, cylindrical, hyaline, unbranched, straight to sinuous. Conidiogenous cells densely aggregated, phiailidic, unbranched, straight or slightly curved, 5.5–10 × 2.0–7.5 μm. Beta conidia filiform, hyaline, straight or slightly curved, aseptate, 17.0–29.5 × 1.0–2.0 μm. Alpha conidia and gamma conidia not observed. Teleomorph not observed.

Culture characteristics.

Colonies covering entire plate after 2 weeks. On PDA with profuse aerial mycelium, white surface, reverse amber. On MEA with fluffy aerial mycelium, dirty white surface, reverse ochreous. On SNA white sparse aerial mycelium, surface and reverse white.

Additional material examined.

China • Guangdong Province, Guangzhou City, Longdong straight street, 23°11'41.02"N, 113°22'8.33"E, 46 m asl, on diseased leaves of Bauhiniavariegata, 8 Aug 2022, Yong Li, Chengbin Wang & Yaquan Zhu, living culture GZ13E.

Notes.

Diaportheguangzhouensis from the present study is phylogenetically close to D.tulliensis (Fig. 2). Diaportheguangzhouensis differs from D.tulliensis in nucleotide sequence data (5/526 in ITS, 9/347 in tef1, 13/711 in tub2) (Crous et al. 2015). In addition, host and distribution data are vital for species identification (D.guangzhouensis inhabiting Bauhiniavariegata in China vs. D.tulliensis inhabiting Theobromacacao in Australia) (Crous et al. 2015).

﻿Discussion

In the current study, phylogenetic analyses based on five combined loci (ITS, cal, his3, tef1 and tub2), as well as morphological characters of the anamorph obtained in culture, revealed D.bauhiniicola and D.guangzhouensis spp. nov. from Bauhiniavariegata, which contributed to our knowledge of the diversity of Diaporthe species in China.

Diaporthepseudobauhiniae (syn. D.chiangraiensis, Chiangraiomycesbauhiniae) was described as a saprobic fungus on branches of Bauhinia sp. in Thailand (Senanayake et al. 2017). D.bauhiniae was introduced from branches of B.purpurea in China (Yang et al. 2021). Hence, a total of four species of Diaporthe have been recorded from the host genus Bauhinia. Phylogenetically, D.bauhiniae belongs to D.varians species complex; D.bauhiniicola belongs to D.arecae species complex; D.guangzhouensis and D.pseudobauhiniae belong to D.sojae species complex (Figs 1, 2) (Norphanphoun et al. 2022). Furthermore, D.guangzhouensis and D.pseudobauhiniae formed different clades in D.sojae species complex (Fig. 2). Morphologically, D.bauhiniicola has larger alpha conidia than D.pseudobauhiniae, but longer alpha conidia than D.bauhiniae (4.5–7.0 × 2.0–3.0 μm in D.bauhiniicola vs. 3–5 × 2–4 μm in D.pseudobauhiniae vs. 7.5–14 × 1.5–3 μm in D.bauhiniae) (Senanayake et al. 2017; Yang et al. 2021). D.guangzhouensis shares similar beta conidia size with D.pseudobauhiniae that are shorter and wider than D.bauhiniae (17.0–29.5 × 1.0–2.0 μm in D.guangzhouensis vs. 18–38 × 1.5–2 μm in D.pseudobauhiniae vs. 25–43 × 1 µm in D.bauhiniae) (Senanayake et al. 2017; Yang et al. 2021). Another species named Phomopsisbauhiniae was recorded on the branches of Bauhiniavariegata in Spain, however, this species was only studied in morphology and has not been combined in Diaporthe (Uecker 1988). Diaporthebauhiniicola has shorter but wider alpha conidia than P.bauhiniae morphologically (Uecker 1988). The molecular analyses are necessary for the species P.bauhiniae based on the ex-type culture in the future.

The initial species concept of Diaporthe based on the assumption of host-specificity, resulted in the introduction of more than 1000 taxa (http://www.indexfungorum.org/). However, more than one species of Diaporthe have been often discovered from the same host (Gomes et al. 2013; Guarnaccia and Crous 2017; Guarnaccia et al. 2020; Guo et al. 2020). For example, D.caryae and an additional 18 Diaporthe species are associated with pear shoot canker in China (Guo et al. 2020); D.sennae and D.sennicola inhabit branches of Sennabicapsularis causing canker diseases (Yang et al. 2017). The current study further supports this phenomenon.

Diaporthe is considered as a species-rich genus. Nevertheless, an emerging perspective posits that the quantity of recognized Diaporthe species may have been substantially overestimated. The D.amygdali species complex has been proven a single species evidenced from the genealogical concordance phylogenetic species recognition principle (GCPSR) and coalescence-based models: general mixed yule-coalescent (GMYC) and poisson tree processes (PTP), with several species becoming synonyms (Hilário et al. 2021b). Similarly, several species in the D.eres species complex such as D.betulae and D.padina were treated as synonyms (Hilário et al. 2021a). A comprehensive study is necessary to clarify species boundaries of Diaporthe in the future. This will help improve our understanding of the species concept within this genus.

Supplementary Material

XML Treatment for Diaporthe bauhiniicola

XML Treatment for Diaporthe guangzhouensis

﻿Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This study was supported by Fundamental Research Funds of CAF (CAFYBB2023PA002).

Author contributions

Conceptualization: LM, YL, NJ. Methodology: YZ. Formal analysis: HX. Investigation: YL. Data Curation: LM, HX. Writing - Original draft: YZ. Writing - Review and Editing: NJ. Visualization: NJ.

Author ORCIDs

Yaquan Zhu https://orcid.org/0000-0002-3296-239X

Han Xue https://orcid.org/0000-0003-0414-6237

Yong Li https://orcid.org/0000-0002-4406-1329

Ning Jiang https://orcid.org/0000-0002-9656-8500

Data availability

All of the data that support the findings of this study are available in the main text.
==== Refs
﻿Reference

Ariyawansa HA Tsai I Wang JY Withee P Tanjira M Lin SR Suwannarach N Kumla J Elgorban AM Cheewangkoon R (2021) Molecular phylogenetic diversity and biological characterization of Diaporthe species associated with leaf spots of Camelliasinensis in Taiwan. Plants 10 (7 ): 1434. 10.3390/plants10071434 34371637
Bai Q Zhai LF Chen XR Hong N Xu WX Wang GP (2015) Biological and molecular characterization of five Phomopsis species associated with pear shoot canker in China. Plant Disease 99 (12 ): 1704–1712. 10.1094/PDIS-03-15-0259-RE 30699520
Caio P Bruno F Carlos AP Robert B (2021) Diaportherosiphthora sp. nov.: Yet another rose dieback fungus. Crop Protection (Guildford, Surrey) 139: 105365. 10.1016/j.cropro.2020.105365
Cao L Luo D Lin W Yang Q Deng X (2022) Four new species of Diaporthe (Diaporthaceae, Diaporthales) from forest plants in China. MycoKeys 91 : 25–47. 10.3897/mycokeys.91.84970 36760894
Carbone I Kohn LM (1999) A Method for designing primer sets for speciation studies in filamentous Ascomycetes. Mycologia 91 (3 ): 553–556. 10.1080/00275514.1999.12061051
Crous PW (2005) Impact of molecular phylogenetics on the taxonomy and diagnostics of fungi. Bulletin OEPP. EPPO Bulletin. European and Mediterranean Plant Protection Organisation 35 (1 ): 47–51. 10.1111/j.1365-2338.2005.00811.x
Crous PW Groenewald JZ Risède JM Simoneau P Hywel-Jones NL (2004) Calonectria species and their Cylindrocladium anamorphs: Species with sphaeropedunculate vesicles. Studies in Mycology 50 : 415–430.
Crous PW Wingfield MJ Roux JJ Richardson DM Strasberg D Shivas RG Alvarado P Edwards J Moreno G Sharma R Sonawane MS Tan YP Altés A Barasubiye T Barnes CW Blanchette RA Boertmann D Bogo A Carlavilla JR Cheewangkoon R Daniel R de Beer ZW Yáñez-Morales MJ Duong TA Fernández-Vicente J Geering ADW Guest DI Held BW Heykoop M Hubka V Ismail AM Kajale SC Khemmuk W Kolařík M Kurli R Lebeuf R Lévesque CA Lombard L Magista D Manjón JL Marincowitz S Mohedano JM Nováková A Oberlies NH Otto EC Paguigan ND Pascoe IG Pérez-Butrón JL Perrone G Rahi P Raja HA Rintoul T Sanhueza RMV Scarlett K Shouche YS Shuttleworth LA Taylor PWJ Thorn RG Vawdrey LL Solano-Vidal R Voitk A Wong PTW Wood AR Zamora JC Groenewald JZ (2015) Fungal Planet description sheets: 371–399. Persoonia 35 (1 ): 264–327. 10.3767/003158515X690269 26823636
Dissanayake AJ Phillips AJL Hyde KD Yan JY Li XH (2017) The current status of species in Diaporthe. Mycosphere : Journal of Fungal Biology 8(5): 1106–1156. 10.5943/mycosphere/8/5/5
Dissanayake AJ Chen YY Liu JK (2020) Unravelling Diaporthe species associated with woody hosts from karst formations (Guizhou) in China. Journal of Fungi (Basel, Switzerland) 6 (4 ): 251. 10.3390/jof6040251 33121032
Dong Z Manawasinghe IS Huang Y Shu Y Phillips AJL Dissanayake AJ Hyde KD Xiang M Luo M (2021) Endophytic Diaporthe associated with Citrusgrandis cv. tomentosa in China. Frontiers in Microbiology 11: e3621. 10.3389/fmicb.2020.609387
Doyle JJ Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12 : 13–15.
Fan XL Yang Q Bezerra JDP Alvarez LV Tian CM (2018) Diaporthe from walnut tree (Juglansregia) in China, with insight of Diaportheeres complex. Mycological Progress 17 (7 ): 1–13. 10.1007/s11557-018-1395-4
Farr DF Castlebury LA Rossman AY (2002a) Morphological and molecular characterization of Phomopsisvaccinii and additional isolates of Phomopsis from blueberry and cranberry in the eastern United States. Mycologia 94 (3 ): 494–504. 10.1080/15572536.2003.11833214 21156520
Farr DF Castlebury LA Rossman AY Putnam ML (2002b) A new species of Phomopsis causing twig dieback of Vacciniumvitisidaea (lingonberry). Mycological Research 106 (6 ): 745–752. 10.1017/S095375620200583X
Glass NL Donaldson GC (1995) Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61 (4 ): 1323–1330. 10.1128/aem.61.4.1323-1330.1995 7747954
Gomes RR Glienke C Videira SIR Lombard L Groenewald JZ Crous PW (2013) Diaporthe: A genus of endophytic, saprobic and plant pathogenic fungi. Persoonia 31 (1 ): 1–41. 10.3767/003158513X666844 24761033
Guarnaccia V Crous PW (2017) Emerging citrus diseases in Europe caused by Diaporthe spp. IMA Fungus 8 (2 ): 317–334. 10.5598/imafungus.2017.08.02.07 29242778
Guarnaccia V Martino I Tabone G Brondino L Gullino ML (2020) Fungal pathogens associated with stem blight and dieback of blueberry in northern Italy. Phytopathologia Mediterranea 59 (2 ): 229–245. 10.14601/Phyto-11278
Guindon S Dufayard JF Lefort V Anisimova M Hordijk W Gascuel O (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML 3.0. Systematic Biology 59 (3 ): 307–321. 10.1093/sysbio/syq010 20525638
Guo YS Crous PW Bai Q Fu M Yang MM Wang XH Du YM Hong N Xu WX Wang GP (2020) High diversity of Diaporthe species associated with pear shoot canker in China. Persoonia 45 (1 ): 132–162. 10.3767/persoonia.2020.45.05 34456374
Hall T (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41 : 95–98.
Hilário S Micael FM Artur A (2021a) Using genealogical concordance and coalescent-based species delimitation to assess species boundaries in the Diaportheeres complex. Journal of Fungi 7 (7 ): 507. 10.3390/jof7070507 34202282
Hilário S Santos L Alves A (2021b) Diaportheamygdali, a species complex or a complex species? Fungal Biology 125(7): 505–518. 10.1016/j.funbio.2021.01.006
Hongsanan S Norphanphoun C Senanayake IC Jayawardena RS Manawasinghe IS Abeywickrama PD Khuna S Suwannarach N Senwanna C Monkai J Hyde KD Gentekaki E Bhunjun CS (2023) Annotated notes on Diaporthe species. Mycosphere 14 (1 ): 918–1189. 10.5943/mycosphere/14/1/12
Huang F Udayanga D Wang X Hou X Mei X Fu Y Hyde KD Li HY (2015) Endophytic Diaporthe associated with Citrus: A phylogenetic reassessment with seven new species from China. Fungal Biology 119 (5 ): 331–347. 10.1016/j.funbio.2015.02.006 25937062
Huang ST Xia JW Zhang XG Sun WX (2021) Morphological and phylogenetic analyses reveal three new species of Diaporthe from Yunnan, China. MycoKeys 78 : 49–77. 10.3897/mycokeys.78.60878 33664613
Hyde KD Nilsson RH Alias SA Ariyawansa HA Blair JE Cai L De Cock AWAM Dissanayake AJ Glockling SL Goonasekara ID Gorczak M Hahn M Jayawardena RS Van Kan JAL Laurence MH Lévesque CA Li XH Liu JK Maharachchikumbura SSN Manamgoda DS Martin FN McKenzie EHC McTaggart AR Mortimer PE Nair PVR Pawłowska J Rintoul TL Shivas RG Spies CFJ Summerell BA Taylor PWJ Terhem RB Udayanga D Vaghefi N Walther G Wilk M Wrzosek M Xu JX Yan JY Zhou N (2014) One stop shop: backbones trees for important phytopathogenic genera: I. Fungal Diversity 67 (1 ): 21–125. 10.1007/s13225-014-0298-1
Jiang N Voglmayr H Piao CG Li Y (2021) Two new species of Diaporthe (Diaporthaceae, Diaporthales) associated with tree cankers in the Netherlands. MycoKeys 85 : 31–56. 10.3897/mycokeys.85.73107 34934385
Katoh K Standley DM (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30 (4 ): 772–780. 10.1093/molbev/mst010 23329690
Maharachchikumbura SSN Hyde KD Jones EBG McKenzie EHC Bhat JD Dayarathne MC Huang SK Norphanphoun C Senanayake IC Perera RH Shang QJ D’souza MJ Hongsanan S Jayawardena RS Daranagama DA Konta S Goonasekara ID Zhuang WY Jeewon R Phillips AJL Abdel-Wahab MA Al-Sadi AM Bahkali AH Boonmee S Boonyuen N Cheewangkoon R Dissanayake AJ Kang J Li QR Liu JK Liu XZ Liu ZY Luangsa-ard JJ Pang KL Phookamsak R Promputtha I Suetrong S Stadler M Wen T Wijayawardene NN (2016) Families of Sordariomycetes. Fungal Diversity 79 (1 ): 1–317. 10.1007/s13225-016-0369-6
Manawasinghe IS Dissanayake AJ Li X Liu M Wanasinghe DN Xu J Zhao W Zhang W Zhou Y Hyde KD Brooks S Yan J (2019) High genetic diversity and species complexity of Diaporthe associated with grapevine dieback in China. Frontiers in Microbiology 10: 1936. 10.3389/fmicb.2019.01936
Marin-Felix Y Hernandez-Restrepo M Wingfield MJ Akulov A Carnegie AJ Cheewangkoon R Gramaje D Groenewald JZ Guarnaccia V Halleen F Lombard L Luangsa-ard J Marincowitz S Moslemi A Mostert L Quaedvlieg W Schumacher RK Spies CFJ Thangavel R Taylor PWJ Wilson AM Wingfield BD Wood AR Crous PW (2019) Genera of phytopathogenic fungi: GOPHY 2. Studies in Mycology 92 (1 ): 47–133. 10.1016/j.simyco.2018.04.002 29997401
Mostert L Crous PW Kang J-C Phillips AJL (2001) Species of Phomopsis and a Libertella sp. occurring on grapevines with specific reference to South Africa: Morphological, cultural, molecular and pathological characterization. Mycologia 93 (1 ): 146–167. 10.1080/00275514.2001.12061286
Norphanphoun C Gentekaki E Hongsanan S Jayawardena R Senanayake C Manawasinghe I Abeywickrama P Bhunjun CS Hyde KD (2022) Diaporthe: Formalizing species-group concepts. Mycosphere 13 (1 ): 752–819. 10.5943/mycosphere/13/1/9
O’Donnell K Cigelnik E (1997) Two divergent intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution 7 (1 ): 103–116. 10.1006/mpev.1996.0376 9007025
Rambaut A Drummond A (2010) FigTree v.1.3.1. Institute of Evolutionary Biology, University of Edinburgh, Edinburgh.
Ronquist F Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19 (12 ): 1572–1574. 10.1093/bioinformatics/btg180 12912839
Rossman AY Adams GC Cannon PF Castlebury LA Crous PW Gryzenhout M Jaklitsch WM Mejia LC Stoykov D Udayanga D Voglmayr H Walker DM (2015) Recommendations of generic names in Diaporthales competing for protection or use. IMA Fungus 6 (1 ): 145–154. 10.5598/imafungus.2015.06.01.09 26203420
Santos JM Phillips AJL (2009) Resolving the complex of Diaporthe (Phomopsis) species occurring on Foeniculumvulgare in Portugal. Fungal Diversity 34 (11 ): 111–125.
Santos L Alves A Alves R (2017) Evaluating multi-locus phylogenies for species boundaries determination in the genus Diaporthe. PeerJ 5: e3120. 10.7717/peerj.3120
Senanayake IC Crous PW Groenewald JZ Maharachchikumbura SSN Jeewon R Phillips AJL Bhat DJ Perera RH Li QR Li WJ Tangthirasunun N Norphanphoun C Karunarathna SC Camporesi E Manawasighe IS Al-Sadi AM Hyde KD (2017) Families of Diaporthales based on morphological and phylogenetic evidence. Studies in Mycology 86 (1 ): 217–296. 10.1016/j.simyco.2017.07.003 28947840
Senanayake IC Jeewon R Chomnunti P Wanasinghe DN Norphanphoun C Karunarathna A Pem D Perera RH Camporesi E McKenzie EHC Hyde KD Karunarathna SC (2018) Taxonomic circumscription of Diaporthales based on multigene phylogeny and morphology. Fungal Diversity 93 (1 ): 241–443. 10.1007/s13225-018-0410-z
Sun W Huang S Xia J Zhang X Li Z (2021) Morphological and molecular identification of Diaporthe species in south-western China, with description of eight new species. MycoKeys 77 : 65–95. 10.3897/mycokeys.77.59852 33519269
Taylor JW (2011) One Fungus = One Name: DNA and fungal nomenclature twenty years after PCR. IMA Fungus 2 (2 ): 113–120. 10.5598/imafungus.2011.02.02.01 22679595
Udayanga D Liu X Crous PW McKenzie EH Chukeatirote E Chukeatirote E Hyde KD (2012a) A multi-locus phylogenetic evaluation of Diaporthe (Phomopsis). Fungal Diversity 56 (1 ): 157–171. 10.1007/s13225-012-0190-9
Udayanga D Liu X McKenzie EH Chukeatirote E Hyde KD (2012b) Multi-locus phylogeny reveals three new species of Diaporthe from Thailand. Cryptogamie. Mycologie 33 (3 ): 295–309. 10.7872/crym.v33.iss3.2012.295
Udayanga D Castlebury LA Rossman AY Hyde KD (2014a) Species limits in Diaporthe: Molecular re-assessment of D.citri, D.cytosporella, D.foeniculina and D.rudis. Persoonia 32(1): 83–101. 10.3767/003158514X679984
Udayanga D Castlebury LA Rossman AY Chukeatirote E Hyde KD (2014b) Insights into the genus Diaporthe: Phylogenetic species delimitation in the D.eres species complex. Fungal Diversity 67 (1 ): 203–229. 10.1007/s13225-014-0297-2
Udayanga D Castlebury LA Rossman AY Chukeatirote E Hyde KD (2015) The Diaporthesojae species complex: Phylogenetic re-assessment of pathogens associated with soybean, cucurbits and other field crops. Fungal Biology 119 (5 ): 383–407. 10.1016/j.funbio.2014.10.009 25937066
Uecker FA (1988) A world list of Phomopsis names with notes on nomenclature, morphology and biology. Morphology and Biology 13 : 1–231.
van Rensburg JCJ Lamprecht SC Groenewald JZ Castlebury LA Crous PW (2006) Characterization of Phomopsis spp. associated with dieback of rooibos (Aspalathuslinearis) in South Africa. Studies in Mycology 55 : 65–74. 10.3114/sim.55.1.65 18490972
Vrandecic K Jurkovic D Cosic J Postic J Riccioni L (2011) First report of cane blight on blackberry caused by Diaportheeres in Croatia. Plant Disease 95 (5 ): 612–612. 10.1094/PDIS-11-10-0860
White TJ Bruns T Lee S Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: A Guide to Methods and Applications 18 : 315–322. 10.1016/B978-0-12-372180-8.50042-1
Wrona CJ Mohankumar V Schoeman MH Tan YP Shivas RG Jeff‐Ego OS Akinsanmi OA (2020) Phomopsis husk rot of macadamia in Australia and South Africa caused by novel Diaporthe species. Plant Pathology 69 (5 ): 911–921. 10.1111/ppa.13170
Yang Q Fan XL Du Z Tian CM (2017) Diaporthe species occurring on Sennabicapsularis in southern China, with descriptions of two new species. Phytotaxa 302 (2 ): 145–155. 10.11646/phytotaxa.302.2.4
Yang Q Fan XL Guarnaccia V Tian CM (2018) High diversity of Diaporthe species associated with dieback diseases in China, with twelve new species described. MycoKeys 39 : 97–149. 10.3897/mycokeys.39.26914
Yang Q Jiang N Tian CM (2020) Three new Diaporthe species from Shaanxi Province, China. MycoKeys 67 : 1–18. 10.3897/mycokeys.67.49483 32425650
Yang Q Jiang N Tian CM (2021) New species and records of Diaporthe from Jiangxi Province, China. MycoKeys 77 : 41–64. 10.3897/mycokeys.77.59999 33519268
Zhang QM Yu CL Li GF Wang CX (2018) First report of Diaportheeres causing twig canker on Zizyphusjujuba (Jujube) in China. Plant Disease 102(7): e1458. 10.1094/PDIS-12-17-1910-PDN
Zhu YQ Ma CY Xue H Piao CG Li Y Jiang N (2023) Two new species of Diaporthe (Diaporthaceae, Diaporthales) in China. MycoKeys 95 : 209–228. 10.3897/mycokeys.95.98969 37251990
