279
Fi e no el species o Cu ula ia (Pleospo ales, Pleospo aceae)
isola ed om u g asses
Jia-Mei Zhao1, Chuan-Xu Peng1, Qiu-Yue Zhang1,2 , De-Wei Li3, Lin Huang1,2
1 College o Fo es y and G assland, Nanjing Fo es y Uni e si y, Nanjing 210037, China
2 Collabo a i e Inno a ion Cen e o Sus ainable Fo es y in Sou he n China, Nanjing 210037, China
3 The Connec icu Ag icul u al Expe imen S a ion Valley Labo a o y, Windso , CT 06095, USA
Co esponding au ho : Lin Huang ([email p o ec ed]); Qiu-Yue Zhang ([email p o ec ed])
Copy igh : © Jia-Mei Zhao e al.
This is an open access a icle dis ibu ed unde
e ms o he C ea i e Commons A ibu ion
License (A ibu ion 4.0 In e na ional – CC BY 4.0).
Resea ch A icle
Abs ac
Cu ula ia, a cosmopoli an ungal genus, occupies a ious ecological niches, bu dis-
plays a p onounced endency o colonise he lea es o plan s. In his s udy, se e al un-
gal isola es wi h simila cha ac e is ics in he genus Cu ula ia we e collec ed om lea
spo s o u g asses (Cynodon dac ylon and Lolium pe enne) in Jiangsu P o ince, China.
Based on he mo phological cha ac e is ics and h ee locus phylogeny o he in e nal
ansc ibed space (ITS) genes, glyce aldehyde-3-phospha e dehyd ogenase (GAPDH)
and ansla ion elonga ion ac o -1 alpha ( e 1), i e new species in Cu ula ia: C. cyno-
don is, C. he bicola, C. loliicola, C. nanjingensis and C. xuanwuensis, a e desc ibed he e-
by. The p esen s udy con ibu es o he unde s anding o species di e si y, axonomy
and phylogeny o Cu ula ia species in China.
Key wo ds: Fungal di e si y, Helmin hospo ioid ungi, hyphomyce es, phylogeny, Pleo-
spo aceae, axonomy
In oduc ion
Tu g ass, de ined as a i icially main ained ege a ion, plays a c ucial ole in
enhancing aes he ics, p e en ing soil e osion and suppo ing ecological bal-
ance in u ban landscapes, spo s and ec ea ional se ings (B aun e al. 2024).
In China, he esea ch on u g ass b eeding s a ed ela i ely la e, wi h only
1–2 a ie ies being egis e ed annually and only abou 20 o hese a ie ies
a e conside ed sui able o lawn applica ions (Lai and Han 2022). As a esul ,
cool-season u g asses, such as Poa p a ensis, Fes uca a undinacea and Lo-
lium pe enne, along wi h wa m-season species like Cynodon dac ylon, a e p e-
dominan ly impo ed (Asano and Ugaki 1994; Inokuma e al. 1998). Tu g ass-
es a e suscep ible o o e 300 ungal pa hogens ha a ec lea es, shea hs
and oo s. No able diseases include b own pa ch, dolla spo s, summe pa ch,
powde y mildew, smu , Py hium bligh , Fusa ium bligh , an h acnose and Cu u-
la ia lea spo s (Couch 1995; Toda e al. 2007; Kamme e e al. 2011).
The genus Cu ula ia Boedijn, belonging o Pleospo aceae, Pleospo ales,
Do hideomyce es, was es ablished and ypi ied by C. luna a (Wakke ) Boedijn
(Boedjin 1933). I is cha ac e ised by he in e cala y and e minal conidiogenous
Academic edi o :
Malgo za a Ruszkiewicz-Michalska
Recei ed:
13 Augus 2025
Accep ed:
20 Oc obe 2025
Published:
21 No embe 2025
Ci a ion: Zhao J-M, Peng C-X, Zhang
Q-Y, Li D-W, Huang L (2025) Fi e no el
species o Cu ula ia (Pleospo ales,
Pleospo aceae) isola ed om
u g asses. MycoKeys 125:
279–305. h ps://doi.o g/10.3897/
mycokeys.125.168614
MycoKeys 125: 279–305 (2025)
DOI: 10.3897/mycokeys.125.168614
280
MycoKeys 125: 279–305 (2025), DOI: 10.3897/mycokeys.125.168614
Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
cells and sympodial conidiopho es. Mos conidia o his genus a e alca e o
cu ed, hough s aigh conidia also occu in some axa (Manamgoda e al.
2015; Fe dinandez e al. 2023; an Vuu en e al. 2024; Ahmadpou e al. 2025).
Cu ula ia species ha e a cosmopoli an dis ibu ion and inhabi di e se niches
as plan pa hogens, endophy es, sap obes o oppo unis ic human pa hogens
(D ans ield 1966; Almague e al. 2012; Manamgoda e al. 2015; Ma in-Felix e
al. 2017). Addi ionally, mos o he species a e epo ed om poaceous hos s
(Smiley e al. 2005; Manamgoda e al. 2012; Tan e al. 2018; Ma in e al. 2020;
T edway e al. 2023). I is a species- ich genus wi h 253 epi he s and a ie ies
in Index Fungo um (h p://www.index ungo um.o g, accessed on 1 July 2025).
Mo phologically, Cu ula ia was ecognised as a genus o en con used wi h
Bipola is Shoemake (Si anesan 1987; Tan e al. 2018; an Vuu en e al. 2024).
These gene a con ain species wi h s aigh o cu ed conidia; he conidia o
Cu ula ia show cu a u e due o disp opo iona ely la ge in e media e cells,
whe eas he cu a u e o Bipola is is con inuous along he en i e leng h o he
conidium. Though, gene ally, Bipola is conidia a e longe han Cu ula ia conid-
ia, hey a e s ill di icul o dis inguish (Manamgoda e al. 2014, 2015; Ma in-Fe-
lix e al. 2020; Ahmadpou e al. 2025). On he o he hand, he sexual mo phs
o hese gene a a e simila and p e iously classi ied as Cochliobolus D echsle
(synonymous wi h Pseudocochliobolus Tsuda, Ueyama & Nishih.), cha ac e ised
by b own o black, globose ascoma a (pseudo hecia), bi unica e and cylind ical
asci and ili o m o lagelli o m, hyaline ascospo es, which a e loosely o ganised
in o a helix o pa allel in he ascus (Si anesan 1987; Manamgoda e al. 2014,
2015). Howe e , i is a e o ind sexual mo phs in na u e (Si anesan 1987).
Due o he di icul y in dis inguishing hese gene a using mo phological cha -
ac e s, species ecogni ion in hese gene a elies on molecula analyses (Ma-
namgoda e al. 2014). Al hough he in e nal ansc ibed space (ITS) egion has
been selec ed as he p ima y ungal ba code ma ke , i has low esolu ion o Cu -
ula ia. The e o e, o specia ion o Cu ula ia, mul i-locus sequence analyses o
ITS and he pa ial gene egions o glyce aldehyde-3-phospha e dehyd ogenase
(GAPDH) and ansla ion elonga ion ac o 1 alpha ( e 1) genes ha e been p o-
posed (Manamgoda e al. 2012; Ma in-Felix e al. 2017, 2020; Tan e al. 2018). Nu-
me ous s udies ha e shown ha Cu ula ia is cu en ly ecognised as a well-de-
ined monophyle ic genus (Manamgoda e al. 2015; Ma in-Felix e al. 2020).
Du ing he ex ensi e in es iga ions conduc ed o collec u g ass pa hogens in
Nanjing, China, se e al Cu ula ia axa exhibi ing ypical cha ac e is ics we e suc-
cess ully isola ed. The p ima y objec i e o his s udy was o de e mine he iden-
i ies o hese newly-collec ed Cu ula ia species h ough mo phological s udies
and phylogene ic analyses, while also elucida ing hei phylogene ic ela ionships.
Ma e ials and me hods
Sample collec ion and po en ial ungal pa hogen isola ion
Samples we e collec ed om lea bligh o g assland in six pa ks ac oss main
u ban dis ic s o Nanjing, Jiangsu P o ince, China. Wi hin each pa k, se en o
en 1 × 1 m quad a s we e su eyed. The comp ehensi e da a o each sampling
si e a e lis ed in Suppl. ma e ial 1: able S1, including speci ic loca ion, su ey
da e, geog aphic coo dina es (la i ude, longi ude), u a ea and u g ass a ie y.
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
Small sec ions (2 × 3 mm) we e cu om he ma gins o in ec ed issues
and su ace s e ilised in 75% alcohol o 30 s, hen in 1% sodium hypochlo i e
(NaOCl) o 90 s, ollowed by h ee inses wi h s e ile wa e (Huang e al. 2016),
hen blo ed d y wi h s e ilised il e pape , placed on Pe i pla es wi h 2% po-
a o dex ose aga (PDA) and 100 mg/l ampicillin and hen cul u ed o 3 days
a 25 °C in he da k. Fungal isola es we e pu i ied wi h he monospo ic isola-
ion me hod desc ibed by using he spo es p oduced wi h liquid cul u es (Li e
al. 2007). Single-spo e isola es we e main ained on PDA pla es. The ob ained
isola es we e s o ed in he Fo es Pa hology Labo a o y a Nanjing Fo es y
Uni e si y. Holo ype specimens o new species om his s udy we e deposi ed
a he China Fo es y Cul u e Collec ion Cen e (CFCC, h ps://c cc.ca .ac.cn/),
Chinese Academy o Fo es y, Beijing, China.
Mo phological analyses
The mo phology o he new species iden i ied in his s udy was analysed,
based on ui ing bodies na u ally o med on lea es and PDA pla es. Mi-
c omo phological s uc u es we e pho og aphed using he Zeiss s e eomi-
c oscope. The shape, colou and size o conidiopho es and conidia we e
obse ed using a ZEISS Axio Scope 5 mic oscope (ZEISS, Ge many) wi h
di e en ial in e e ence con as (DIC) op ics. Fo each s uc u e, 30 mea-
su emen s we e made. Colou s we e de e mined using Ko ne up and Wan-
sche (1978). Pho o pla es we e made by Adobe Pho oshop 2021. Fo spo e
measu emen s, 30 spo es we e andomly selec ed. The esul s a e p esen -
ed as maximum and minimum alues (in pa en heses), along wi h he ange
exp essed as he (min.–) X–SD–X + SD (–max.).
Phylogene ic analyses
Genomic DNA o 26 isola es was ex ac ed using a modi ied CTAB me hod
(Damm e al. 2008). The ungal plugs o each isola e we e g own on he PDA
pla es o 5 days and hen collec ed in a 2 ml ube. Then, 500 µl o chlo o o m
and 500 µl o hexadecyl ime hyl ammonium b omide (CTAB) ex ac ion bu e
(0.2 M T is, 1.4 M NaCl, 20 mM EDTA, 0.2 g/l CTAB) we e added in o he ubes,
which we e placed in a shake a 30 °C a 200 pm o 1.5 h. The mix u e was
cen i uged a 15,800 × g o 10 min. Then, 300 µl o he supe na an was ans-
e ed in o a new ube and 600 µl o 100% e hanol we e added. The suspension
was cen i uged a 15,800 × g o 5 min. A ha poin , 600 µl o 75% e hanol
we e added in o he p ecipi a e. The suspension was cen i uged a 15,800 ×
g o 5 min and he supe na an was disca ded. The DNA pelle was d ied and
e-suspended in 30 µl ddH2O. The quali y o he ex ac ed DNA was assessed
using a mic o olume spec opho ome e .
To ampli y he ITS, GAPDH and e 1 loci, he ollowing p ime pai s we e
used: ITS1/ITS4 (Whi e e al. 1990), gpd1/gpd2 (Be bee e al. 1999) and
e 1-983F/ e 1-2218R (Rehne and Buckley 2005), espec i ely. The p ime
sequences a e lis ed in Suppl. ma e ial 1: able S2. The pa ame e s o PCR
p o ocol we e as ollows: 94 °C o 4 min, ollowed by 35 cycles o 30 s a
94 °C, 45 s a 55–60 °C and 45 s a 72 °C, which we e used in a ouch down
PCR (Ko bie and Ma ick 2008). A inal ex ension s ep was conduc ed o
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
10 min a 72 °C. The p ime s we e syn hesised by Sangon Bio ech, Nanjing,
Jiangsu P o ince, China. The ampli ied p oduc s we e sequenced in Shang-
hai Jie Li Bio echnology Co., L d.
To include a mo e comp ehensi e ange o species, we chose speci ic
genes/ egions o ou s udy, based on esea ch by Ahmadpou e al. (2025).
These we e as ollows: ITS + GAPDH + e 1 o Cu ula ia (Table 1). The da ase
was hen aligned sepa a ely using MAFFT .74 (h p://ma .cb c.jp/alignmen /
se e /) (Ka oh e al. 2019) wi h he G-INS-I i e a i e e inemen algo i hm and
op imised manually in BioEdi .7.0.5.3 (Hall 1999). The sepa a e alignmen s
we e hen conca ena ed using PhyloSui e .1.2.2 (Zhang e al. 2020a).
Phylogene ic ees we e in e ed using bo h Maximum Likelihood (ML)
and Bayesian In e ence (BI) app oaches, implemen ed in RAxML .8.2.10
(S ama akis 2014) and M Bayes 3.2.6 (Ronquis e al. 2012), espec i ely.
Fo he ML analysis, s a is ical suppo alues we e ob ained by using apid
boo s apping wi h 1000 eplica es, wi h de aul se ings o o he pa am-
e e s. Fo BI, he op imal pa i ioning scheme and subs i u ion model we e
selec ed wi h ModelFinde (Kalyaanamoo hy e al. 2017) ia i s “g eedy”
algo i hm. B anch leng hs we e linked ac oss pa i ions and selec ed by
AICc. Fou Ma ko Chain Mon e Ca lo chains (one cold) we e cons uc ed
o 5,000,000 gene a ions, wi h sampling e e y 1000 gene a ions. Following
he bu n-in phase ( i s 25% o sampled ees), he disca ded ees we e ex-
cluded and pos e io p obabili ies (BPP) in he majo i y ule consensus ee
we e calcula ed om he emaining ees.
Phylogene ic ees we e isualised by using FigT ee e sion 1.4.4 (Rambau
2018). B anches ha ecei ed boo s ap suppo s o ML (≥ 50%) and BPP (≥
0.90) we e conside ed as signi ican ly suppo ed.
Resul s
Symp oms in he ield
The symp oms o disease caused by Cu ula ia species isible in he ield a e
shown in Fig. 1. On diseased plan s, he lea es exhibi p og essi e yellowing
and become densely speckled wi h yellowish-b own o black nec o ic lesions.
As he disease ad ances, hese spo s coalesce in o la ge con luen pa ches,
ul ima ely causing comple e lea wil ing and chlo osis. Unde humid condi ions,
he lesion su aces de elop dis inc black mould-like s uc u es, composed o
ungal mycelia and conidia. These symp oms closely esemble hose induced
by Cu ula ia s ains on u g ass species, as p e iously documen ed in pa ho-
logical s udies (Tomaso-Pe e son e al. 2016).
Phylogeny
In his s udy, he conca ena ed (ITS + GAPDH + e 1) da ase included sequenc-
es om 217 s ains, ep esen ing 188 species o Cu ula ia. Sequences o
Bipola is maydis (Y. Nisik. & C. Miyake) Shoemake and used as an ou g oup
ollowed Ahmadpou e al. (2025). The inal alignmen consis ed o 2,006 cha -
ac e s (ITS: 471; GAPDH: 552; e 1: 983), including gaps. Fo Bayesian In e -
ence (BI) analysis, he mos app op ia e models o each locus we e con i med
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
Table 1. Cul u es, specimens and NCBI accession numbe s included in his s udy. T indica es ex- ype s ains.
Species Isola e/Cul u e
collec ion Hos /Subs a um Coun y GenBank accessions
ITS GAPDH e 1
Bipola is maydis CBS 137271 TZea mays USA AF071325 KM034846 KM093794
Cu ula ia ae ia CBS 294.61 TAi B azil HF934910 HG779148 –
C. a inis CBS 154.34 TUnknown Indonesia KJ909780 KM230401 KM196566
C. akaii CBS 318.86 TUnknown Japan LT631340 LT715797 –
C. akaiiensis BRIP 16080 TUnknown India KJ415539 KJ415407 KJ415453
C. alco nii MFLUCC 10-0703 TZea mays Thailand JX256420 JX276433 JX266589
C. alge iensis CBS 150506 TZea mays Alge ia OP218257 OP223173 OP223186
C. ame icana UTHSC 08-3414 THuman ankle USA HE861833 HF565488 –
C. and opogonis CBS 186.49 TAnd opogon na dus Indonesia LT631354 LT715835 –
C. angsiewkeeae BRIP 72449aTScle ia sp. Aus alia OK638993 OK655929 OK655924
C. annelliconidiopho i CGMCC 3.19352 TSaccha um o icina um China MN215641 MN264077 MN263935
C. asia ica MFLUCC 10-0711 TPanicum sp. Thailand JX256424 JX276436 JX266593
C. au an ia USJCC-0096 TZea mays S i Lanka OQ275217 OQ269628 OQ332409
C. aus aliensis BRIP 12044 TO yza sa i a Aus alia KJ415540 KJ415406 KJ415452
C. aus alis BRIP 12521 TSpo obolus ca oli Aus alia KJ415541 KJ415405 KJ415451
C. aus iaca CBS 102694 THuman nasal ca i y Aus ia MN688802 MN688829 MN688856
C. bannonii BRIP 16732 TJacquemon ia amni olia USA KJ415542 KJ415404 KJ415450
C. beasleyi BRIP 10972 TChlo is gayana Aus alia MH414892 MH433638 MH433654
C. beasleyi BRIP 15854 Lee sia hexand a Aus alia MH414893 MH433639 MH433655
C. bee bu umensis BRIP 12942 TE ag os is bahiensis Aus alia MH414895 MH433634 MH433657
C. boe emae IMI 164633 TPo ulaca ole acea India MH414911 MH433641 –
C. bo e iae CBS 859.73 TVolcanic ash soil Chile LT631355 LT715838 –
C. bo h iochloae BRIP 12522 TBo h iochloa bladhii Aus alia KJ415543 KJ415403 KJ415449
C. boudouaouensis CBS 150515 TZea mays Alge ia OP218258 OP223175 OP223187
C. b achyspo a CBS 186.50 TSoil Ja a HG778983 KM061784 KM230405
C. buchloes CBS 246.49 TBuchloe dac yloides USA KJ909765 KM061789 KM196588
C. cac i o a CBS 580.74 Membe o Cac aceae Republic o Su iname MN688803 MN688830 MN688857
C. canadensis CBS 109239 TO e win e ed g ass Canada MN688804 MN688831 MN688858
C. ca icae-papayae CBS 135941 TCa ica papaya India LT631350 LT715816 –
C. caspica IRAN 4275CTEleocha is sp. I an PP593896 PP661368 PP661351
C. caspica FCCUU 1103 Cype us sp. I an PP593897 PP661369 PP661352
C. caspica FCCUU 1104 Fimb is ylis sp. I an PP593898 PP661370 PP661353
C. chiangmaiensis CPC 28829 TZea mays Thailand MF490814 MF490836 MF490857
C. chiang aiensis MFLUCC 22-0091 TSoil Thailand OP581428 OP859013 OP859017
C. chlamydospo a UTHSC 07-2764 THuman oenail USA HG779021 HG779151 –
C. chonbu iensis MFLUCC 16-0375 TPandanus sp. Thailand MH275055 MH412747 –
C. chuasooengiae BRIP 72482a TScle ia sp. Aus alia OK638997 OK655933 –
C. cla a a BRIP 61680b O yza u ipogon Aus alia KU552205 KU552167 KU552159
C. coa esiae BRIP 24261 TLi chi chinensis Aus alia MH414897 MH433636 MH433659
C. coicicola HSAUP 990901 Coix lac yma-jobi China AB453880 – –
C. coicis CBS 192.29 TCoix lac yma-jobi Japan HF934917 HG779130 JN601006
C. coimba o ensis SZMC 22225 THuman co nea India MN628310 MN628306 MN628302
C. colb anii BRIP 13066 TC inum zeylanicum Aus alia MH414898 MH433642 MH433660
C. como iensis CBS 110673 Unknown Unknown LT631357 LT715841 –
C. c assisep a a CBS 503.90 TPlan ma e ial Nige ia LT631310 LT715882 MN688859
C. c us acea BRIP 13524 TSpo obolus sp. Indonesia KJ415544 KJ415402 KJ415448
C. cu culiginis YZU 181230 Cu culigo capi ula a China MK507796 MK507794 MK507795
C. cymbopogonis CBS 419.78 TYucca sp. Ne he lands HG778985 HG779129 –
C. cynodon is SCM16-22TLea es o Cynodon dac ylon China PV973428 PV995188 PV995204
C. cynodon is YZ26-27 Lea es o Cynodon dac ylon China PV973429 PV995189 PV995206
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Species Isola e/Cul u e
collec ion Hos /Subs a um Coun y GenBank accessions
ITS GAPDH e 1
C. cynodon is XW6-23 Lea es o Cynodon dac ylon China PV973430 PV995190 PV995205
C. cype i IRAN 4274C TCype us sp. I an PP593899 PP661371 PP661354
C. cype i FCCUU 1101 Fimb is ylis sp. I an PP593900 PP661372 PP661355
C. cype i FCCUU 1102 Isolepis sp. I an PP593901 PP661373 PP661356
C. dac yloc enicola CPC 28810 TDac yloc enium aegyp ium Thailand MF490815 MF490837 MF490858
C. dac yloc enii BRIP 12846 TDac yloc enium adulans Aus alia KJ415545 KJ415401 KJ415447
C. dac yloc enii 7938-9 Eu eka bluebe y China AF158106 AF081376 –
C. deigh onii CBS 537.70 TSo ghum ulga e Denma k LT631356 LT715839 –
C. dese icola CBS 151410 TS ipag os is cilia a Namibia ON074985 ON355399 ON355360
C. de e mina a CGMCC 3.19340 TSaccha um o icina um China MN215653 MN264088 MN263947
C. eleusinicola USJCC-0005 TEleusine co acana S i Lanka MT262877 MT393583 MT432925
C. ellip i o mis CGMCC 3.19351 TSaccha um o icina um China MN215656 MN264091 MN263950
C. ellisii CBS 193.62 TAi Pakis an JN192375 JN600963 JN601007
C. e ag os idicola BRIP 12538 TE ag os is pilosa Aus alia MH414899 MH433643 MH433661
C. e ag os idis CBS 189.48 TSo ghum sp. Ja a HG778986 HG779154 –
C. alsiluna a CGMCC 3.19329 TSaccha um o icina um China MN215660 MN264093 MN263954
C. lexuosa CGMCC 3.19447 TSaccha um o icina um China MN215663 MN264096 MN263957
C. ankliniae BRIP 72476aTSo ghum imo ense Aus alia OK638995 OK655931 OK655926
C. ase ae BRIP 64708aTBo h iochloa insculp a Aus alia OM809867 OM721558 OM714552
C. genicula a CBS 187.50 TAnd opogon so ghum Indonesia KJ909781 KM083609 KM230410
C. gladioli CBS 210.79 Gladiolus sp. Romania LT631345 LT715802 –
C. gobabebensis CBS 149140 TS ipag os is cilia a Namibia ON074797 ON355381 ON355347
C. gobabebensis CMW-IA 6921 S ipag os is cilia a Namibia ON332848 ON355373 ON355344
C. g aminicola BRIP 23186aTA is ida ing a a Aus alia JN192376 JN600964 JN601008
C. he bicola JHS14-23 Lea es o Lolium pe enne China PV973431 PV995191 PV995207
C. he bicola YYH23-17 Lea es o Cynodon dac ylon China PV973432 PV995192 PV995208
C. he bicola XW1-15TLea es o Cynodon dac ylon China PV973433 PV995193 PV995209
C. guangxiensis CGMCC 3.19330 TSaccha um o icina um China MN215667 MN264100 MN263961
C. gudauskasii DAOM 165085 Unknown Unknown AF071338 AF081393 –
C. ha eyi BRIP 57412 TT i icum aes i um Aus alia KJ415546 KJ415400 KJ415446
C. hawaiiensis BRIP 11987 TO yza sa i a USA KJ415547 KJ415399 KJ415445
C. he e opogonicola BRIP 14579 THe e opogon con o us India KJ415548 KJ415398 KJ415444
C. he e opogonis CBS 284.91 THe e opogon con o us Aus alia KJ415549 JN600969 JN601013
C. hominis UTHSC 09-464 THuman co nea USA HG779011 HG779106 –
C. homomo pha CBS 156.60 TAi USA JN192380 JN600970 JN601014
C. huamulaniae BRIP 10936a TAi Aus alia OR130931 OR135531 OR135532
C. hus oniae BRIP 72486a THe e opogon i iceus Aus alia OK638999 OK655935 OK655928
C. inaequalis CBS 102.42 T Soil F ance KJ922375 KM061787 KM196574
C. in e media CBS 334.64 A ena e sicolo USA HG778991 HG779155 –
C. in e media XY-3 Eu eka bluebe y China OQ300329 OQ338159 OQ338160
C. i anica IRAN 3487C TBougain illea spec abilis I an MT551122 MN266487 MN266490
C. ischaemi CBS 630.82 TIschaemum indicum Solomon Islands HG778992 HG779131 –
C. jolio cu ieae BRIP 14448a TT i icum aes i um Aus alia OQ917073 OQ889556 OQ889557
C. kenpeggii BRIP 14530 TT i icum aes i um Aus alia MH414900 MH433644 MH433662
C. khuzes anica CBS 144736 TA iplex len i o mis I an MH688044 MH688043 –
C. khuzes anica SCUA-11C-2 A iplex len i o mis I an MH688046 MH688045 –
C. kusanoi CBS 137.29 TE ag os is majo Japan JN192381 LT715862 KM196592
C. laming onensis BRIP 12259 TMic olaena s ipoides Aus alia MH414901 MH433645 MH433663
C. loliicola XW9-5-1TLea es o Lolium pe enne China PV973443 PV995201 PV995217
C. loliicola XW9-5-2 Lea es o Lolium pe enne China PV973441 PV995202 PV995218
C. loliicola XW9-5-3 Lea es o Lolium pe enne China PV973442 PV995203 PV995219
C. lolii CMAA 1785 TLolium mul i lo um B azil MT849336 MT889299 MT881706
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Species Isola e/Cul u e
collec ion Hos /Subs a um Coun y GenBank accessions
ITS GAPDH e 1
C. lona ensis CBS 140569 TLona lake India KT315408 KY007019 –
C. luna a CBS 730.96 THuman lung biopsy USA JX256429 JX276441 JX266596
C. lycope sici S ain 11 Solanum lycope sicum Egyp KY883347 KY883345 –
C. malina CBS 131274 TZoysia ma ella USA JF812154 KP153179 KR493095
C. manamgodae CGMCC 3.19446 TSaccha um o icina um China MN215677 MN264110 MN263971
C. ma aisii CBS 149143 TSoil Namibia OR471647 ON355439 OR486044
C. mebaldsii BRIP 12900 TCynodon ans aalensis Aus alia MH414902 MH433647 MH433664
C. mebaldsii CMW-IA 6956 S ipag os is cilia a Namibia ON644443 ON661549 –
C. mic ai ae BRIP 17068aTMic ai a subuli olia Aus alia OM421618 OM373204 OM373205
C.mic opus CBS 127235 TPaspalum no a um USA HE792934 LT715859 –
C. mic ospo a GUCC 6272 THippeas um s ia um China MF139088 MF139106 MF139115
C. millisiae BRIP 71718a TCype us a oma icus Aus alia OK661031 OK636415 OK636413
C. miyakei CBS 197.29 TE ag os is pilosa Japan KJ909770 KM083611 KM196568
C. mo ingae CPC 38873 TMo inga o ali olia Namibia MW175363 MW173105 –
C. mosaddeghii IRAN 3131CTSyzygium cumini I an MG846737 MH392155 MH392152
C. muehlenbeckiae CBS 144.63 TMuehlenbeckia sp. India HG779002 HG779108 –
C. namibensis CBS 149144 TS ipag os is cilia a Namibia ON074819 ON355384 ON355350
C. nanjingensis YZ25-10-2-1TLea es o Cynodon dac ylon China PV973438 PV995198 PV995214
C. nanjingensis YZ25-10-2-2 Lea es o Cynodon dac ylon China PV973439 PV995199 PV995215
C. nanjingensis YZ25-10-2-3 Lea es o Cynodon dac ylon China PV973440 PV995200 PV995216
C. nanningensis GUCC 11005 TCymbopogon ci a us China MH885321 MH980005 MH980011
C. nee gaa dii BRIP 12919 TO yza sa i a Ghana KJ415550 KJ415397 KJ415443
C. neoindica IMI 129790 TB assica nig a India MH414910 MH433649 MH433667
C. nico iae BRIP 11983 TSoil Alge ia KJ415551 KJ415396 KJ415442
C. nodosa CPC 28800 TDigi a ia cilia is Thailand MF490816 MF490838 MF490859
C. nuci e ae YzU 231509 Nelumbo nuci e a China OR888819 PP066825 PP792703
C. nuci e ae YzU 231510 Nelumbo nuci e a China OR888818 PP066826 PP792704
C. nodosa CPC 28801 –Thailand MF490817 MF490839 MF490860
C. nodulosa CBS 160.58 Eleusine indica USA JN601033 JN600975 JN601019
C. nodulosa IRAN 4804C Eleusine indica I an PP593892 PP661366 –
C. nuci e ae YzU 231509 Nelumbo nuci e a China OR888819 PP066825 PP792703
C. nuci e ae YzU 231510 Nelumbo nuci e a China OR888818 PP066826 PP792704
C. o yzae CBS 169.53 TO yza sa i a Vie nam KP400650 HG779156 KM196590
C. o yzae IRAN 5044C Unknown I an PP593893 PP661367 –
C. o yzae-sa i ae CBS 127725 TO yza sa i a A gen ina MN688808 MN688835 MN688863
C. o a iicola CBS 470.90 TE ag os is in e up a Aus alia MN688809 MN688836 –
C. pallescens CBS 156.35 TAi Indonesia KJ922380 KM083606 KM196570
C. palmicola MFLUCC 14-0404 TAcoelo haphe w igh ii Thailand MF621582 – –
C. pandanicola MFLUCC 15-0746 TPandanus sp. Thailand MH275056 MH412748 MH412763
C. panici-maximi USJCC-0006 TPanicum maximum S i Lanka MN044757 MN053040 MN053009
C. papendo ii CBS 308.67 TAcacia ka oo Sou h A ica KJ909774 KM083617 KM196594
C. pa a e uculosa FMR 17656 TSoil Mexico LR736641 LR736646 LR736649
C. pa e eae CBS 198.87 TT i icum du um A gen ina MN688810 MN688837 MN688864
C. pennise i CBS 528.70 Unknown Unknown MH859833 LT715840 –
C. pe o idis CBS 350.90 TPe o is a a Aus alia HG778995 HG779138 KM230407
C. pe e sonii BRIP 14642 TDac yloc enium aegyp ium Aus alia MH414905 MH433650 MH433668
C. phaeospa a CGMCC 3.19448 TSaccha um o icina um China MN215686 MN264118 MN263980
C. pisi CBS 190.48 TPisum sa i um Canada KY905678 KY905690 KY905697
C. plan a um CGMCC 3.19342 TSaccha um o icina um China MN215688 MN264120 MN263982
C. pla zii BRIP 27703b TCench us clandes inum Aus alia MH414906 MH433651 MH433669
C. poly a a CGMCC 3.19338 TSaccha um o icina um China MN215691 MN264123 MN263984
C. p asadii CBS 143.64 TJasminum sambac India KJ922373 KM061785 KM230408
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Species Isola e/Cul u e
collec ion Hos /Subs a um Coun y GenBank accessions
ITS GAPDH e 1
C. p o ube ans CGMCC 3.19360 TSaccha um o icina um China MN215693 MN264125 MN263986
C. p o ube a a CBS 376.65 TDeschampsia lexuosa Sco land KJ922376 KM083605 KM196576
C. pseudob achyspo a CPC 28808 TEleusine indica Thailand MF490819 MF490841 MF490862
C. pseudocla a a CBS 539.70 TO yza sa i a Denma k MN688817 MN688844 MN688869
C. pseudoellisii CBS 298.80 TSo ghum bicolo Sudan MN688818 MN688845 MN688870
C. pseudoin e media CBS 553.89 TCul i a ed pas u e soil B azil MN688819 MN688846 MN688871
C. pseudoluna a UTHSC 09-2092 THuman nasal sinus USA HE861842 HF565459 –
C. pseudop o ube a a CBS 385.69 TSoil unde Thuja occiden alis Canada MN688821 MN688848 MN688873
C. pseudo obus a UTHSC 08-3458 Human nasal sinus USA HE861838 HF565476 –
C. adicicola CGMCC 3.19327 TSaccha um o icina um China MN215699 MN264131 MN263992
C. adici- oliigena CGMCC 3.19328 TSaccha um o icina um China MN215695 MN264127 MN263988
C. a enelii BRIP 13165 TSpo obolus e ilis Aus alia JN192386 JN600978 JN601024
C. eesii BRIP 4358 TAi Aus alia MH414907 MH433637 MH433670
C. icha diae BRIP 4371 TRicha dia b asiliensis Aus alia KJ415555 KJ415391 KJ415438
C. obus a CBS 624.68 TDichan hium annula um USA KJ909783 KM083613 KM196577
C. ouhanii CBS 144674 TSyngonium ellozianum I an KX139030 MG428694 MG428687
C. ouhanii CN022H5 S ipag os is cilia a Namibia ON074966 ON355388 ON355353
C. yleyi BRIP 12554 TSpo obolus c ebe Aus alia KJ415556 KJ415390 KJ415437
C. saccha icola CGMCC 3.19344 TSaccha um o icina um China MN215701 MN264133 MN263994
C. saccha i-o icina um CGMCC 3.19331 TSaccha um o icina um China MN215705 MN264137 MN263998
C. senegalensis CBS 149.71 Unknown Nige ia HG779001 HG779128 –
C. sesu ii CGMCC 3.9578 TSesu ium po ulacas um China EF175940 – –
C. shahidcham anensis IRAN 3133CTSoil I an MH550084 MH550083 –
C. sichuanensis BN9 Ai China MH483998 – –
C. siddiquii CBS 196.62 TAi Pakis an MN688823 MN688850 –
C. simmonsii USJCC-0002 TPanicum maximum S i Lanka MN044753 MN053011 MN053005
C. soli CBS 222.96 TSoil Papua New Guinea KY905679 KY905691 KY905698
C. so ghina BRIP 15900 TSo ghum bicolo Aus alia KJ415558 KJ415388 KJ415435
C. spici e a CBS 274.52 Soil Spain JN192387 JN600979 JN601023
C. spici e a AT-102 co n No he n Alge ia OP218259 OP223176 OP223188
C. spo obolicola BRIP 23040b TSpo obolus aus alasicus Aus alia MH414908 MH433652 MH433671
C. s ipag os idicola CBS 149150 TS ipag os is cilia a Namibia ON332838 ON355415 ON355368
C. s eno aph i BRIP 71303 TS eno aph um secunda um Aus alia MZ681952 MZ695824 MZ695819
C. subpapendo ii CBS 656.74 TSoil Egyp KJ909777 KM061791 KM196585
C. su oniae FMR 10992 THuman leg wound USA HE861828 HF565479 LR736651
C. abaci YzU 221481 Nico iana abacum China OR888817 PP066824 OR818398
C. abaci YzU 221482 Nico iana abacum China PP601257 PP779503 PP779504
C. abaci YzU 221481 Nico iana abacum China OR888817 PP066824 OR818398
C. abaci YzU 221482 Nico iana abacum China PP601257 PP779503 PP779504
C. amilnaduensis SZMC 22226 THuman co nea India MN628311 MN628307 MN628303
C. anzanica IMI 507176 TCype us a oma icus Tanzania MW396857 MW388669 –
C. emple oniae BRIP 72453a THypa henia hi a Aus alia OK638994 OK655930 OK655925
C. hailandicum MFLUCC 15-0747 TPandanus sp. Thailand MH275057 MH412749 MH412764
C. ibuli CBS 126975 TT ibulus e es is Sou h A ica MN688825 MN688852 MN688875
C. i olii ICMP 6149 Se a ia glauca New Zealand KM230395 KM083607 JX266600
C. ipogonis BRIP 12375 TT ipogon lolii o mis Aus alia JN192388 JN600980 JN601025
C. opicalis BRIP 14834 TCo ea a abica India KJ415559 KJ415387 KJ415434
C. sudae ATCC 44764 TChlo is gayana Japan KC424596 KC747745 KC503940
C. ube cula a CBS 146.63 TZea mays India JX256433 JX276445 JX266599
C. umbilici o mis CGMCC 3.19346 TSaccha um o icina um China MN215711 MN264142 MN264004
C. uncina a CBS 221.52 TO yza sa i a Vie nam HG779024 HG779134 –
C. a iabilis CPC 28815 TChlo is ba ba a Thailand MF490822 MF490844 MF490865
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Species Isola e/Cul u e
collec ion Hos /Subs a um Coun y GenBank accessions
ITS GAPDH e 1
C. e uci o mis CBS 537.75 Vanellus miles NewZealand HG779026 HG779133 –
C. e ucosa CBS 422.93 Ai Cuba MN688826 MN688853 MN688876
C. e uculosa CBS 150.63 Punica g ana um India KP400652 KP645346 KP735695
C. e uculosa NCK 1675 Human skin Taiwan LC802641LC802651 LC802661
C. idyodayana USJCC-0029 TO yza sa i a S i Lanka OQ275234 OQ269645 OQ332413
C. ie namensis FMR 17659 Tuniden i ied dead lea es Vie nam LR736642 LR736644 LR736647
C. wa abe ensis BRIP 14817 TDac yloc enium aegyp ium Aus alia MH414909 MH433653 MH433672
C. xishuangbannaensis KUMCC 17-0185 TPandanus ama ylli ollus Thailand MH275058 MH412750 MH412765
C. xuanwuensis XW4-33 Lea es o Cynodon dac ylon China PV973434 PV995194 PV995210
C. xuanwuensis XW9-10 Lea es o Cynodon dac ylon China PV973435 PV995195 PV995211
C. xuanwuensis ZS19-14TLea es o Cynodon dac ylon China PV973436 PV995196 PV995212
C. xuanwuensis ZS19-15 Lea es o Cynodon dac ylon China PV973437 PV995197 PV995213
C. yamadana COAD 359 Cype us o undus B azil MN954705 –MT008260
Cu ula ia sp. IRAN 4273C Saccha um o icina um I an PP593902 PP661374 PP661357
Cu ula ia sp. IRAN 3500C Saccha um o icina um I an PP593903 PP661375 PP661358
Figu e 1. Disease symp oms associa ed wi h u g ass samples (Cynodon dac ylon and Lolium pe enne) in his s udy.
A–D. Symp oms documen ed a he si e o collec ion; E–H. De ailed iew in he collec ed Lolium pe enne lea es; I–L.
De ailed iew in he collec ed Cynodon dac ylon lea es.
using ModelFinde (The selec ed models we e SYM + I + G4 o ITS, GTR + F +
I + G4 o GAPDH and GTR + F + I + G4 o e 1). Bayesian analysis esul ed in a
nea ly cong uen opology wi h an a e age s anda d de ia ion o spli equen-
cies as 0.011855 o ML analysis and, hus, only he ML ee is p o ided (Fig. 2).
ML boo s ap suppo alues (BS) ≥ 50% and Bayesian pos e io p obabili ies
(BPP) ≥ 0.90 a e indica ed on he b anches.
Phylogene ic analyses e ealed i e new dis inc lineages o Cu ula ia
amongs he examined isola es (Fig. 2), ul ima ely leading o he ecogni ion o
i e new species, based on mo phological cha ac e is ics and mul i-locus phy-
logeny (ITS, GAPDH and e 1). Sequences o h ee s ains (SCM16-22, XW6-23,
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Figu e 5. Cu ula ia loliicola (XW9-5-1, ype). A–C. Colony on PDA (A), OA (B) and CMA (C), espec i ely in 5 days a e
incuba ion a 25 °C; D. Spo ula ion on PDA; E–F. Conidiogenous cell and conidiopho e; G–K. Conidia, conidiogenous cell
and conidiopho e; L. Conidia. Scale ba s: 100 μm (D); 50 μm (H); 10 μm (E–G, I–L).
295
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g eyish-g een and he edge is da k yellow. On OA medium, he colonies each up
o 47 mm in diam. The edge is nea , co ony appea ance, he colony su ace is
g ey, ae ial hyphae a e lu y and e e se is black. On CMA medium, he colonies
a e co ony and each 32 mm in diam. The edge is i egula and he cen e is black.
Addi ional ma e ials examined. China • Jiangsu P o ince, Nanjing, Xuanwu
Dis ic , Xuanwu Lake Pa k, isola ed om lea bligh o Lolium pe enne, 5 June
2024, Jia-Mei Zhao and Lin Huang, XW9-5-2, XW9-5-3.
No es. Phylogene ically, Cu ula ia loliicola o ms a dis inc , s ongly sup-
po ed lineage sis e o C. in e media (Fig. 2). Mo phologically, C. loliicola e-
sembles C. in e media in p oducing 3-sep a e conidia, bu di e s in conidial
and conidiopho e size and o namen a ion. Cu ula ia in e media possesses
longe conidia (33–37 s. 23–29 μm in leng h). In addi ion, C. in e media dis-
inc ly di e s om C. loliicola by i s longe conidiopho es which can each up o
800 μm, while he conidiopho es o C. loliicola show no basal enla gemen and
a e semi- o mac o-nema ous, up o 192 μm (Ahmadpou e al. 2012). Fu he -
mo e, i has also been epo ed ha i can cause lea spo s o Cynodon dac y-
lon, bluebe ies and o he plan s (Couch 1995; Li e al. 2019; Cheng e al. 2022;
Kong e al. 2024). In addi ion, he no el species esembles Cu ula ia g aminis
Meng Zhang & T.Y. Zhang, desc ibed om g ass hos s in China, pa icula ly in
conidial mo phology, whe e he second and hi d sep a om he base a e o en
da ke pigmen ed. Howe e , C. g aminis di e s om C. loliicola in ha ing longe
conidia (29–46 s. 23–29 μm, Zhang and Zhang (2007)).
Cu ula ia nanjingensis Lin Huang, Jia-Mei Zhao & D.W. Li, sp. no .
Fig. 6
MycoBank No: 860703
Holo ype. China • Jiangsu P o ince, Nanjing, Jianye Dis ic , Yuzui We land Pa k,
31°97'13"N, 118°65'54"E, isola ed om lea bligh o Cynodon dac ylon, 29 Aug
2024. Holo ype: CFCC 72723 is a li ing specimen being main ained ia lyophilisa-
ion a he China Fo es y Cul u e Collec ion Cen e (CFCC). Ex- ype (YZ25-10-2-
1) is main ained a he Fo es Pa hology Labo a o y, Nanjing Fo es y Uni e si y.
E ymology. The name e e s o he ci y, Nanjing whe e he holo ype was
collec ed.
Desc ip ion. Asexual mo ph on PDA: Hyphae 3–4 μm wide, subhyaline o pale
b own, hin- and smoo h-walled, sep a e, b anched. Conidiopho es mononema ous,
semi- o mac onema ous, sep a e, a ising singly o equen ly in g oups, s aigh o
lexuous, genicula e owa ds he apex, unb anched, smoo h-walled, subhyaline o
da k b own, (113–)163–279(–384) × (3–)4–6 μm (mean ± SD = 221 ± 58 × 5 ± 1
μm). Conidiogenous cells mono- o poly e ic, p oli e a ing sympodially, in eg a -
ed, e minal o in e cala y, subcylind ical o i egula ly swollen, yellowish-b own,
smoo h, 6–14(–18) × 4–6 μm (mean ± SD = 10 ± 4 × 5 ± 1 μm). Conidia s aigh
o cu ed, ellip ical o luna e, smoo h, 2–3-eusep a e, median cells unequally en-
la ged, pigmen a ion in ensi ied o da k b own and median sep um hickened, api-
cal and basal cells subhyaline o pale b own, (13–)16–20(–22) × (7–)9–13(–14)
μm (mean ± SD = 18 ± 2 × 11 ± 2 μm); ge mina ion mono- o bipola . Hila 2–3
μm wide, inconspicuous o sligh ly conspicuous, sligh ly hickened and da kened.
Chlamydospo es p esen , in e cala y, smoo h-walled, soli a y o g ouped in chain,
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Figu e 6. Cu ula ia nanjingensis (YZ25-10-2-1, ype). A–C. Colony on PDA (A), OA (B) and CMA (C), espec i ely in 5
days a e incuba ion a 25 °C; D. Spo ula ion on PDA; E–K. Conidia, conidiogenous cells and conidiopho e; L. Chlamydo-
spo es; M–N. conidia. Scale ba s: 100 μm (D); 10 μm (H–M).
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
subglobose o oblong, 8–12(–13) × (6–)7–9(–10) μm (mean ± SD = 10 ± 2 × 8 ± 1
μm, n = 20). Mic oconidia no obse ed. Sexual mo ph: Unde e mined.
Cul u e cha ac e is ics. Colonies on PDA each 46 mm diam. a e 5 days
a 25 °C in he da k. Round colonies, egula ma gin, la wi hou bulge, g ey-
ish-black a spo ula ing zone, whi e a aging zone. The cen e o he e e se
is g eyish-g een and he edge is yellowish-whi e. On OA medium, he colonies
a e up o 51 mm in diam., wi h egula ma gin, smoo h su ace, black on bo h
on and e e se sides. On CMA medium, he diame e is up o 49 mm and he
mo phology is simila o ha on OA medium.
Addi ional ma e ials examined. China • Jiangsu P o ince, Nanjing, Jianye
Dis ic , Yuzui We land Pa k, 29 Augus 2024, isola ed om lea bligh o Cyno-
don dac ylon, Jia-Mei Zhao and Lin Huang, YZ25-10-2-2, YZ25-10-2-3.
No es. Phylogene ically, Cu ula ia nanjingensis is alloca ed o a s ongly sup-
po ed lineage (100/0.98) in Cu ula ia and ela ed o C. chiang aiensis and C.
simmonsii (Fig. 2). Howe e , C. chiang aiensis can be easily dis inguished om
C. nanjingensis by i s smalle conidiopho es (50–150 × 2–7 μm s. 163–279 ×
4–6 μm, Yasan hika e al. (2023)). Cu ula ia simmonsii di e s om C. nanjin-
gensis by i s longe conidia (21–27 μm s. 16–20 μm, Fe dinandez e al. (2021)).
Addi ionally, C. nanjingensis can p oduce chlamydospo es, while his ea u e is
no obse ed in C. chiang aiensis and C. simmonsii. Fu he mo e, C. chiang aien-
sis and C. simmonsii ha e only been spo adically epo ed. The o me has been
epo ed o be isola ed om soil; he la e was isola ed om he lea lesions o
Panicum maximum in S i Lanka (Fe dinandez e al. 2021; Yasan hika e al. 2023).
Cu ula ia xuanwuensis Lin Huang, Jia-Mei Zhao & D.W. Li, sp. no .
Fig. 7
MycoBank No: 860704
Holo ype. China • Jiangsu P o ince, Nanjing, Xuanwu Dis ic , Zhongshan Spo s
Pa k, 32°05'03"N, 118°86'69"E, isola ed om lea bligh o Cynodon dac ylon, 3
July 2024. Holo ype: CFCC 72729 is a li ing specimen being main ained ia lyoph-
ilisa ion a he China Fo es y Cul u e Collec ion Cen e (CFCC). Ex- ype (ZS19-
14) is main ained a he Fo es Pa hology Labo a o y, Nanjing Fo es y Uni e si y.
E ymology. The epi he name a e he Xuanwu Dis ic whe e he holo ype
o he ungus was collec ed.
Desc ip ion. Asexual mo ph on PDA: Hyphae 2–4 μm wide, subhyaline o
pale b own, hin- and smoo h-walled hyphae, sep a e, b anched. Conidiopho es
mononema ous, semi- o mac onema ous, sep a e, a ising singly o equen ly in
g oups, s aigh o lexuous, genicula e a uppe pa , unb anched, smoo h, pale
b own o b own, pale owa ds he apex, (49–)60–180(–239) × (3–)4–6(–7) μm
(mean ± SD = 120 ± 60 × 5 ± 1 μm). Conidiogenous cells mono- o poly e ic, p o-
li e a ing sympodially, in eg a ed, e minal o in e cala y, subcylind ical, yellow-
ish-b own, smoo h o sligh ly e uculose, wi h hickened and da kened sca s,
(6–)7–11(–14) × 4–6(–7) μm (mean ± SD = 9 ± 2 × 5 ± 1 μm). Conidia s aigh ,
pale b own o da k golden b own, smoo h, ellipsoidal, 2–3-eusep a e, (14–)18–
22(–24) × (7–)8–10 μm (mean ± SD = 20 ± 2 × 9 ± 1 μm); ge mina ion mono- o
bipola . Hila 2–3 μm wide, non-p o ube an , hickened and da kened. Chlamydo-
spo es and mic oconidia we e no obse ed. Sexual mo ph no obse ed.
298
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
Figu e 7. Cu ula ia xuanwuensis (ZS19-14, ype). A–C. Colony on PDA (A), OA (B) and CMA (C), espec i ely in 5 days
a e incuba ion a 25 °C; D. Spo ula ion on PDA; E. Conidiogenous cell and conidiopho e; F–K. Conidia, conidiogenous
cells and conidiopho es; L. Conidia. Scale ba s: 100 μm (D); 20 μm (H–I, K); 10 μm (E–G, J, L).
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
Cul u e cha ac e is ics. Colonies on PDA each 44 mm diame e a e 5 days
a 25 °C in he da k. Round colonies wi h an en i e ma gin, g eyish-black a spo -
ula ing zone, la and whi e a aging zone, co ony appea ance. The cen e o
he e e se is g eyish-g een and he edge is whi e. On OA medium, he colonies
a ain up o 51 mm in diam. The edge is nea , co ony appea ance, he su ace
o he colony is g ey-black, he edge is whi e and he ae ial hyphae a e lu y.
Re e se side is g eyish-black in he cen e and whi e on he edges. On CMA
medium, he colony is up o 49 mm in diam., wi h an i egula ma gin, he on
and back sides a e black and he su ace is smoo h.
Addi ional ma e ials examined. China • Jiangsu P o ince, Nanjing, Xuanwu
Dis ic , Zhongshan Spo s Pa k, 3 July 2024, isola ed om lea bligh o Cy-
nodon dac ylon, Jia-Mei Zhao and Lin Huang, ZS19-15; Xuanwu Lake Pa k, 5
June 2024, isola ed om lea bligh o Cynodon dac ylon, Jia-Mei Zhao and Lin
Huang, XW4-33, XW9-10.
No es. Ou phylogene ic analyses e eal ha Cu ula ia xuanwuensis o ms
a s ongly suppo ed (100/1) lineage and sis e s o C. aus aliensis. Howe e ,
hese species a e eadily dis inguishable in mo phology. C. aus aliensis di e s
om C. xuanwuensis by i s longe conidia (15–40 μm s. 18–22 μm), wi h hick-
e and mo e conspicuous conidial sep a and la ge conidiopho es (95–205 ×
3–7 μm s. 60–180 × 4–6 μm, La o e (2015)).
Discussion
Species iden i ica ion wi hin he genus Cu ula ia adi ionally elies on mo -
phological cha ac e is ics, encompassing shape and leng h o conidiopho es
and conidia, as well as he p esence o s uc u es like hilum, s oma, chlam-
ydospo es and mic oconidia (Tan e al. 2014, 2018; Manamgoda e al. 2015;
Ma in-Felix e al. 2020). Howe e , eliable species dis inc ion is challenging due
o many o e lapping mo phological cha ac e s (Si anesan 1987; Alco n 1988;
Manamgoda e al. 2012, 2015). S udies ha e epo ed mul iple isola ion o Cu -
ula ia species om a single plan in di e en geog aphic loca ions, u he
con i ming he complexi y o Cu ula ia species delinea ion (Ahmadpou e al.
2014; Heida ian e al. 2020; Fe dinandez e al. 2021, 2023). The e o e, phylo-
gene ic in e ence, based on DNA sequence da a, is essen ial. Fu he mo e, ITS
sequences alone a e insu icien o accu a ely di e en ia e indi idual species
and phylogene ic analyses using mul i-locus ITS + GAPDH + e 1 genomic loci
p o ide he highes esolu ion o species bounda ies amongs Cu ula ia spe-
cies (Be bee e al. 1999; Kiss e al. 2020; an Vuu en e al. 2024; Ahmadpou
e al. 2025; Guo e al. 2025). Consequen ly, he mode n axonomic sys em o
Cu ula ia is based on mo phology and mul i-locus phylogene ic analyses. In
his s udy, i e new species o Cu ula ia a e iden i ied in Jiangsu P o ince, Chi-
na: C. cynodon is, C. he bicola, C. loliicola, C. nanjingensis and C. xuanwuensis.
Cu ula ia species exhibi a wide ange o ecological oles, including plan , ani-
mal o human pa hogens, as well as epiphy es, sap obes o endophy es, p edom-
inan ly associa ed wi h cul i a ed ce eals (Manamgoda e al. 2015; Ma in-Felix e
al. 2020; Fa e al. 2021; Ahmadpou e al. 2025). Amongs hem, as he phy o-
pa hogens, Cu ula ia species we e p ima ily associa ed wi h g asses o he Poa-
ceae, such as majo g ains like ice, whea and co n, as well as o namen al plan s
like u g asses (Smiley e al. 2005; Zhang e al. 2020b; T edway e al. 2023; Ram
300
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Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
e al. 2024). In his s udy, based on mul i-locus phylogene ic analyses and mo -
phological a ibu es, i e new Cu ula ia species we e desc ibed om o namen-
al Poaceae plan s in China. Cu ula ia cynodon is, C. xuanwuensis and C. nanjin-
gensis we e only isola ed om Cynodon dac ylon and C. cynodon is was he mos
equen ly isola ed species in his s udy, including 11 s ains. In addi ion, o o he
species desc ibed in his s udy, C. loliicola was ound only in he lea es o Lolium
pe enne, while C. he bicola was isola ed om hese wo g ass plan s.
Cu ula ia is a cosmopoli an genus, wi h mo e han 30 i s epo s on di e -
en plan s in China, India and Pakis an ound in li e a u e om 2010 o he p es-
en (Zhang e al. 2020b). I is e iden ha Cu ula ia species ha e main ained
a close associa ion wi h plan di e si y. Al hough his s udy e ealed associa-
ions be ween speci ic Cu ula ia species and pa icula subs a es (Cynodon
dac ylon and Lolium pe enne), b oade sampling would help be e unde s ood
hei biology, subs a e p e e ences, and dis ibu ion pa e ns.
Collec i ely, his s udy con ibu es o a be e unde s anding o he di e si y
o Cu ula ia isola es associa ed wi h Poaceae hos s. Fu he , his s udy p o-
ides aluable insigh s o plan pa hologis s, mycologis s, ag onomis s and
en i onmen al scien is s wi h ac ionable insigh s o enhance conse a ion and
managemen s a egies o key Poaceae o namen als o c ops.
Addi ional in o ma ion
Con lic o in e es
The au ho s ha e decla ed ha no compe ing in e es s exis .
E hical s a emen
No e hical s a emen was epo ed.
Use o AI
No use o AI was epo ed.
Funding
Pos g adua e Resea ch & P ac ice Inno a ion P og am o Jiangsu P o ince (G an /
Awa d Numbe : KYCX23_1225), Qing Lan P ojec , and P io i y Academic P og am De-
elopmen o Jiangsu Highe Educa ion Ins i u ions (PAPD).
Au ho con ibu ions
Sampling, Jia-Mei Zhao and Chuan-Xu Peng; ungal isola ion and sequencing Jia-Mei
Zhao and Chuan-Xu Peng; phylogene ic analysis, Jia-Mei Zhao, Qiu-Yue Zhang and De-
Wei Li; desc ip ion, Jia-Mei Zhao; w i ing – o iginal d a p epa a ion, Jia-Mei Zhao and
Qiu-Yue Zhang; w i ing – e iew and edi ing, Jia-Mei Zhao, Chuan-Xu Peng, Huang Lin,
Qiu-Yue Zhang, De-Wei Li; supe ision, Huang Lin. All au ho s ha e ead and ag eed o
he published e sion o he manusc ip .
Au ho ORCIDs
Jia-Mei Zhao h ps://o cid.o g/0009-0005-6748-8682
Qiu-Yue Zhang h ps://o cid.o g/0000-0001-9458-3566
De-Wei Li h ps://o cid.o g/0000-0002-2788-7938
Lin Huang h ps://o cid.o g/0000-0001-7536-0914
301
MycoKeys 125: 279–305 (2025), DOI: 10.3897/mycokeys.125.168614
Jia-Mei Zhao e al.: Fi e no el species o Cu ula ia isola ed om u g asses
Da a a ailabili y
All o he da a ha suppo he indings o his s udy a e a ailable in he main ex o
Supplemen a y In o ma ion.
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