Research Article
Print
Research Article
Morphological and phylogenetic analyses reveal three new species of Apiospora in China
expand article infoShuji Li‡, Cheng Peng‡, Rong Yuan‡, Chengming Tian‡
‡ Beijing Forestry University, Beijing, China

Corresponding author: Chengming Tian ( chengmt@bjfu.edu.cn )

Academic editor: Ning Jiang
© 2023 Shuji Li, Cheng Peng, Rong Yuan, Chengming Tian.
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.
Citation: Li S, Peng C, Yuan R, Tian C (2023) Morphological and phylogenetic analyses reveal three new species of Apiospora in China. MycoKeys 99: 297-317. https://doi.org/10.3897/mycokeys.99.108384
Open Access

Abstract

Species of Apiospora are distributed worldwide as endophytes, pathogens and saprobes. In this study, we analysed Apiospora strains isolated from diseased leaves in Yunnan Province and dead culms in Shaanxi Province, China and we identified fungal species based on multi-locus phylogeny of ITS, LSU, tef1 and tub2 genes, along with the morphological characters, host and ecological distribution. Analyses revealed three new species, namely A. coryli sp. nov., A. lophatheri sp. nov. and A. oenotherae sp. nov. and one known species A. arundinis. Illustrations and descriptions of the four taxa are provided, along with comparisons with closely-related taxa in the genus.

Key words

Apiosporaceae, Ascomycota, morphology, phylogeny, taxonomy

Introduction

Species in Apiospora are distributed worldwide, primarily in temperate and tropical regions. These fungi can be found in various habitats, including soil, plant materials and insect exoskeletons (Pintos and Alvarado 2021). Many species of Apiospora are associated with plants as endophytic or saprophytic taxa and some can be important plant pathogens (Crous and Groenewald 2013; Wang et al. 2018; Kwon et al. 2021). In recent years, researchers have continuously discovered new Apiospora species in China (Wang et al. 2018; Senanayake et al. 2020, 2023; Feng et al. 2021; Liu et al. 2023).

Apiospora, the type genus of Apiosporaceae, was recognised and established by Saccardo (1875) with A. montagnei as the type species. For a long time, Apiospora was believed to be the sexual state of the genus Arthrinium (Ellis 1965; Samuels et al. 1981; Crous and Groenewald 2013). However, Ellis (1965) synonymised several other asexual genera with basauxic conidiogenesis under Arthrinium, such as Papularia, which was considered the asexual morph of Apiospora by von Höhnel (1919), Petrak (1925) and Hudson (1960, 1963). The asexual morph of Apiospora and Arthrinium are difficult to differentiate, based on morphology alone and the morphological relationships between Arthrinium and Apiospora have been hotly debated since Ellis (1965).

With the help of molecular phylogeny, Apiospora and Arthrinium were initially categorised in their own family Apiosporaceae (Hyde et al. 1998). Later, Crous and Groenewald (2013) considered that Apiospora was actually the sexual form of Arthrinium and both genera aligned to form a monophyletic clade. Following the principle of one fungi, one name policy (Hawksworth et al. 2011), the older name Arthrinium was recommended for use in unitary nomenclature (Réblová et al. 2016). However, due to several names with comparable sexual morphs to those of Arthrinium described as A. montagnei, the exact identity of A. montagnei remained uncertain (Hudson et al. 1976; Pintos et al. 2019; Pintos and Alvarado 2021). With the availability of sequence data of A. montagnei, Pintos and Alvarado (2022) revealed that Apiospora and Arthrinium are distinct genera. With most Apiospora species sharing similar morphologies, molecular phylogenetic information is necessary for accurate species identification (Pintos and Alvarado 2022).

The aim of the present study is to research new Apiospora samples found in western China, including one known species of A. arundinis and three new species and to describe them, based on morphological characters and phylogeny inferred from the combined ITS, LSU, tef1 and tub2 sequences dataset. To identify and compare these species with morphologically similar and phylogenetically related species, thorough analyses have been conducted.

Materials and methods

Sample collection and fungal isolation

Diseased leaves with dried dark brown spots of Oenothera biennis and Lophatherum gracile, as well as diseased leaves with white round patches and black cracks of Brunfelsia brasiliensis were collected from two locations in Yunnan Province: Lincang City (1547 m elevation; 23°52'12"N, 100°4'12"E) and Xishuangbanna City (763 m elevation; 22°1'48"N, 100°52'48"E). Dead plant culms of Corylus yunnanensis were collected in Ankang City (1683 m elevation; 33°26'37"N, 108°26'4"E), located in Shaanxi Province. All samples were placed in paper bags and transported to the laboratory for isolation. The samples were surface-sterilised by being exposed to 75% ethanol for one minute, followed by 1.25% sodium hypochlorite for three minutes, then another minute of exposure to 75% ethanol. The samples were then rinsed with distilled water for two minutes and dried on sterile filter paper. The affected portions of the leaves were excised into 0.5 × 0.5 cm fragments using a sterile razor blade. The fragments were then placed on to potato dextrose agar plates (PDA; containing 200 g potatoes, 20 g dextrose and 20 g agar per litre). The plates were incubated at a temperature of 25 °C to obtain pure cultures. All specimens were deposited at the Museum of Beijing Forestry University (BJFC) and all cultures were preserved at the China Forestry Culture Collection Center (CFCC).

Morphological observation

The morphology of the isolates was examined by analysing sporulating axenic cultures cultivated on PDA in darkness at 25 °C. After a 7-day incubation period, colony diameters were measured and colony characters were recorded. Slide mounts were prepared in lactic acid or water, obtained from colonies sporulating on PDA. Observations were conducted using a Leica DM 2500 dissecting microscope (Wetzlar, Germany) and a Nikon Eclipse 80i compound microscope, equipped with differential interference contrast (DIC) illumination. Images were captured with a Nis DS-Ri2 camera and processed using the Nikon Nis Elements F4.30.01 software. For measurement purposes, 50 conidiogenous cells and conidia were randomly selected. Conidial length was measured from the base of the basal cell to the base of the apical appendage, while conidial width was measured at its widest point. Taxonomic novelties were deposited in MycoBank (http://www.mycobank.org).

DNA extraction, PCR amplification and phylogenetic analyses

Genomic DNA was extracted from colonies grown on PDA using a cetyltrimethylammonium bromide (CTAB) method (Doyle and Doyle 1990). The extracted DNA products were stored at -20 °C until analysis. Four different loci were targeted for sequencing, including the nrDNA internal transcribed spacer regions 1 and 2 with the intervening 5.8S subunit (ITS), a partial sequence of the large subunit nrDNA subunit (LSU), a partial sequence of the translation elongation factor 1-alpha gene (tef1) and a partial sequence of the beta-tubulin gene (tub2). They were all amplified with the primer pairs and polymerase chain reaction (PCR) programme listed in Table 1.

Table 1.
Download as
CSV
XLSX

Gene regions and respective primer pairs used in the study.

Locus PCR primers PCR: thermal cycles: (Annealing temperature in bold) Reference
ITS ITS1/ITS4 (94 °C: 30 s, 55 °C: 30 s, 72 °C: 45 s) × 35 cycles White et al. 1990
LSU LR0R/LR5 (94 °C: 30 s, 48 °C: 50 s, 72 °C: 1 min 30 s) × 35 cycles Cubeta et al. 1991
tef1 EF1-728F/EF2 (95 °C: 30 s, 51 °C: 30 s, 72 °C: 1 min) × 35 cycles O’Donnell et al. 1998; Carbone and Kohn 1999
tub2 Bt-2a/Bt-2b (95 °C: 30 s, 56 °C: 30 s, 72 °C: 1 min) × 35 cycles Glass and Donaldson 1995

The PCR products were assayed by electrophoresis in 2% agarose gels. Amplified PCR products were sent to a commercial sequencing provider (Tsingke Biotechnology Co. Ltd., Beijing, China). The quality of the chromatograms was verified and nucleotide sequences were assembled using SeqMan v.7.1.0. Reference sequences from related publications (Wang et al. 2018; Pintos and Alvarado 2021; Samarakoon et al. 2022; Liu et al. 2023) were retrieved from the National Center for Biotechnology Information (NCBI; https://www.ncbi.nlm.nih.gov). Sequences were aligned on the web server using MAFFT at the web server (http://mafft.cbrc.jp/alignment/server) (Katoh et al. 2019) and further corrected manually utilising MEGA 7.0.21 (Kumar et al. 2016).

The phylogenetic analyses of the combined loci were performed using Maximum Likelihood (ML) and Bayesian Inference (BI) methods. To implement ML, RAxMLHPC BlackBox 8.2.10 (Stamatakis 2014) was used on the CIPRES Science Gateway portal (https://www.phylo.org) employing a GTR GAMMA substitution model with a total of 1000 bootstrap replicates. The Bayesian posterior probabilities (BPP) were determined by Markov Chain Monte Carlo (MCMC) sampling in MrBayes v.3.2.6 (Ronquist et al. 2012). Six simultaneous Markov chains were run for 1 million generations starting from random trees, sampling trees every 100th generation. To ensure accuracy, 25% of aging samples were discarded, running until the average standard deviation of the split frequencies dropped below 0.01. The phylogram was visualised in FigTree v.1.3.1 (http://tree.bio.ed.ac.uk/software) and edited using Adobe Illustrator CS5 (Adobe Systems Inc., USA). The newly-generated nucleotide sequences were deposited in GenBank (Table 2).

Table 2.
Download as
CSV
XLSX

Isolates and GenBank accession numbers used in the phylogenetic analyses.

Species Isolate/Strain Host/ Substrate Origin GenBank accession numbers
ITS LSU tef1 tub2
Apiospora acutiapica KUMCC 20-0210 (Type) Bambusa bambos China MT946343 MT946339 MT947360 MT947366
A. agari KUC 21333 (Type) Agarum cribrosum Korea MH498520 MH498440 MH544663 MH498478
A. aquatica MFLU 18-1628 (Type) Submerged wood China MK828608 MK835806 NA NA
A. arctoscopi KUC 21331 (Type) Egg of Arctoscopus japonicus Korea MH498529 MH498449 MN868918 MH498487
A. arundinis CBS 10612 Unkown substrate Germany KF144883 KF144927 KF145015 KF144973
LX 1918 Saccharum officinarum China MW534386 NA MW584370 MZ090019
CFCC 58977 Brunfelsia brasiliensis China OR125562 OR133584 OR139968 OR139976
LS 107 Brunfelsia brasiliensis China OR125563 OR133585 OR139969 OR139977
A. aurea CBS 24483 (Type) Air Spain AB220251 KF144935 KF145023 KF144981
A. balearica CBS 145129 (Type) Poaceae Spain MK014869 MK014836 MK017946 MK017975
A. bambusae ICPM 6889 (Type) Bamboo China MK014874 MK014841 MK017951 MK017980
A. bambusicola MFLUCC 20-0144 (Type) Schizostachyum brachycladum Thailand MW173030 MW173087 MW183262
A. biserialis CGMCC 320135 (Type) Bamboo China MW481708 MW478885 MW522938 MW522955
A. camelliae-sinensis LC 5007 (Type) Camellia sinensis China KY494704 KY494780 KY705103 KY705173
A. chromolaenae MFLUCC 17-1505 (Type) Chromolaena odorata Thailand MT214342 MT214436 MT235802 NA
A. chiangraiense MFLUCC 21-0053 (Type) Bamboo Thailand MZ542520 MZ542524 NA MZ546409
A. cordylinae GUCC 10027 (Type) Cordyline fruticosa China MT040106 NA MT040127 MT040148
A. coryli CFCC 58978 (Type) Corylus yunnanensis China OR125564 OR133586 OR139974 OR139978
CFCC 58979 Corylus yunnanensis China OR125565 OR133587 OR139975 OR139979
A. cyclobalanopsidis CGMCC 320136 (Type) Cyclobalanopsidis glauca China MW481713 MW478892 MW522945 MW522962
A. descalsii CBS 145130 (Type) Ampelodesmos mauritanicus Spain MK014870 MK014837 MK017947 MK017976
A. dichotomanthi LC 4950 (Type) Dichotomanthus tristaniaecarpa China KY494697 KY494773 KY705096 KY705167
A. dongyingensis SAUCC 0302 (Type) Bamboo China OP563375 OP572424 OP573264 OP573270
A. esporlensis CBS 145136 (Type) Phyllostachys aurea Spain MK014878 MK014845 MK017954 MK017983
A. euphorbiae IMI 285638b Bambusa Bangladesh AB220241 AB220335 NA AB220288
A. fermenti KUC21289 (Type) Seaweed Korea MF615226 MF615213 MH544667 MF615231
A. gaoyouense CFCC 52301 (Type) Phragmites australis China MH197124 NA MH236793 MH236789
A. garethjonesii JHB004 (Type) Bamboo China KY356086 KY356091 NA NA
A. gelatinosa HKAS 111962 (Type) Bamboo China MW481706 MW478888 MW522941 MW522958
A. guiyangensis HKAS 102403 (Type) Poaceae China MW240647 MW240577 MW759535 MW775604
A. guizhouensis LC 5322 (Type) Air in karst cave China KY494709 KY494785 KY705108 KY705178
A. hainanensis SAUCC 1681 (Type) Bamboo China OP563373 OP572422 OP573262 OP573268
A. hispanicum IMI 326877 (Type) Maritime sand Spain AB220242 AB220336 NA AB220289
A. hydei CBS 114990 (Type) Bambusa tuldoides China KF144890 KF144936 KF145024 KF144982
A. hyphopodii MFLUCC 15-0003 (Type) Bamboo China KR069110 NA NA NA
A. ibericum AP 10118 (Type) Arundo donax Portugal MK014879 MK014846 MK017955 MK017984
A. intestini CBS 135835 (Type) Gut of grasshopper India KR011352 MH877577 KR011351 KR011350
A. italicum CBS 145138 (Type) Arundo donax Italy MK014880 MK014847 MK017956 MK017985
A. jatrophae CBS 134262 (Type) Jatropha podagrica India JQ246355 NA NA NA
A. jiangxiensis LC 4577 (Type) Maesa sp. China KY494693 KY494769 KY705092 KY705163
A. kogelbergensis CBS 113333 (Type) Restionaceae South Africa KF144892 KF144938 KF145026 KF144984
A. koreanum KUC 21332 (Type) Egg of Arctoscopus japonicus Korea MH498524 MH498444 MH544664 MH498482
A. lageniformis KUC 21686 (Type) Phyllostachys nigra Korea ON764020 ON787759 ON806624 ON806634
A. locuta-pollinis LC 11683 (Type) Brassica campestris China MF939595 NA MF939616 MF939622
A. longistroma MFLUCC 11-0481 (Type) Bamboo Thailand KU940141 KU863129 NA NA
A. lophatheri CFCC 58975 (Type) Lophatherum gracile China OR125566 OR133588 OR139970 OR139980
CFCC 58976 Lophatherum gracile China OR125567 OR133589 OR139971 OR139981
A. malaysiana CBS 102053 (Type) Macaranga hullettii stem colonised by ants Malaysia KF144896 KF144942 KF145030 KF144988
A. marianiae AP18219 (Type) Phleum pratense Spain ON692406 ON692422 ON677180 ON677186
A. marii CBS 49790 (Type) Atmosphere, pharmaceutical excipients, home dust and beach sands Spain MH873913 KF144947 KF145035 KF144993
A. marinum KU 21328 (Type) Seaweed China MH498538 MH498458 MH544669 MH498496
A. mediterranea IMI 326875 (Type) Air Spain AB220243 AB220337 NA AB220290
A. minutisporum 17E-042 (Type) Soil Korea LC517882 NA LC518889 LC518888
A. montagnei AP 301120 (Type) Arundo micrantha Spain ON692408 ON692424 ON677182 ON67718
A. mori MFLU 18-2514 (Type) Morus australis China MW114313 MW114393 NA NA
A. mukdahanensis MFLUCC 22-0056 (Type) Bambusoideae Thailand OP377735 OP377742 OP381089 NA
A. multiloculata MFLUCC 21-0023 (Type) Bambusae Thailand OL873137 OL873138 NA OL874718
A. mytilomorpha DAOM 214595 (Type) Andropogon India KY494685 NA NA NA
A. neobambusae LC 7106 (Type) Bamboo China KY494718 KY494794 KY806204 KY705186
A. neochinensis CFCC 53036 (Type) Fargesia qinlingensis China MK819291 NA MK818545 MK818547
A. neogarethjonesii HKAS 102408 (Type) Bambusae China MK070897 MK070898 NA NA
A. neosubglobosa JHB007 (Type) Bamboo China KY356090 KY356095 NA NA
A. obovatum LC4940 (Type) Lithocarpus China KY494696 KY494772 KY705095 KY705166
A. oenotherae CFCC 58972 (Type) Oenothera biennis China OR125568 OR133590 OR139972 OR139982
LS 395 Oenothera biennis China OR125569 OR133591 OR139973 OR139983
A. ovata CBS 115042 (Type) Arundinaria hindsii China KF144903 KF144950 KF145037 KF144995
A. paraphaeosperma MFLUCC13-0644 (Type) Bambusa Thailand KX822128 KX822124 NA NA
A. phragmitis CBS 135458 (Type) Phragmites australis Italy KF144909 KF144956 KF145043 KF145001
A. phyllostachydis MFLUCC 18-1101 (Type) Phyllostachys heteroclada China MK351842 MH368077 MK340918 MK291949
A. piptatheri CBS 145149 (Type) Piptatherum miliaceum Spain MK014893 MK014860 MK017969 NA
A. pseudomarii GUCC 10228 (Type) Aristolochia debilis China MT040124 NA MT040145 MT040166
A. pseudohyphopodii KUC 21680 (Type) Phyllostachys pubescens Korea ON764026 ON787765 ON806630 ON806640
A. pseudoparenchymaticum LC 7234 (Type) Bamboo China KY494743 KY494819 KY705139 KY705211
A. pseudorasikravindrae KUMCC 20-0208 (Type) Bambusa dolichoclada China MT946344 NA MT947361 MT947367
A. pseudosinensis CBS 135459 (Type) Bamboo Netherlands KF144910 KF144957 KF145044 NA
A. pseudospegazzinii CBS 102052 (Type) Macaranga hullettii Malaysia KF144911 KF144958 KF145045 KF145002
A. pterosperma CPC 20193 (Type) Lepidosperma gladiatum Australia KF144913 KF144960 KF145046 KF145004
A. pusillisperma KUC 21321 (Type) Seaweed Korea MH498533 MH498453 MN868930 MH498491
A. qinlingense CFCC 52303 (Type) Fargesia qinlingensis China MH197120 NA MH236795 MH236791
A. rasikravindrae NFCCI 2144 (Type) Soil in karst cave China JF326454 NA NA NA
A. sacchari CBS 21230 Phragmites australis Korea KF144919 KF144965 KF145050 KF145008
A. saccharicola CBS 19173 Air Netherlands KF144920 KF144966 KF145051 KF145009
A. sargassi KUC21228 (Type) Sargassum fulvellum Korea KT207746 KT207696 MH544677 KT207644
A. sasae CBS 146808 (Type) Sasa veitchii Netherlands MW883402 MW883797 MW890104 MW890120
A. septata CGMCC 320134 (Type) Bamboo China MW481711 MW478890 MW522943 MW522960
A. serenensis IMI 326869 (Type) Food, pharmaceutical excipients, atmosphere and home dust Spain AB220250 AB220344 NA AB220297
A. setariae CFCC 54041 (Type) Setaria viridis China MT492004 NA NA NA
A. setostroma KUMCC 19-0217 (Type) Bambusoideae China MN528012 MN528011 MN527357 NA
A. sichuanensis HKAS 107008 (Type) Poaceae China MW240648 MW240578 MW759536 MW775605
A. sorghi URM 93000 (Type) Sorghum bicolor Brazil MK371706 NA NA MK348526
A. sphaerosperma CBS114314 (Type) Hordeum vulgare Iran KF144904 KF144951 KF145038 KF144996
A. stipae CBS 146804 (Type) Stipa gigantea Spain MW883403 MW883798 MW890082 MW890121
A. subglobosa MFLUCC 11-0397 (Type) Bamboo Thailand KR069112 KR069113 NA NA
A. subrosea LC7292 (Type) Bamboo China KY494752 KY494828 KY705148 KY705220
A. taeanensis KUC21322 (Type) Seaweed Korea MH498515 MH498435 MH544662 MH498473
A. thailandica MFLUCC 15-0202 (Type) Rotten wood China KU940145 KU863133 NA NA
A. vietnamense IMI 99670 (Type) Citrus sinensis Vietnam KX986096 KX986111 NA KY019466
A. xenocordella CBS 47886 (Type) Soil from roadway Zimbabwe KF144925 KF144970 KF145055 KF145013
A. yunnana MFLUCC 15-0002 (Type) Bamboo China KU940147 KU863135 NA NA
Arthrinium crenatum CBS 146353B (Type) Grass France MW208931 MW208861 MW221917 MW221923

Notes: Strains in this study are marked in bold. NA = not available.

Results

Phylogeny

The combined ITS, LSU, tef1 and tub2 dataset comprised 99 strains, including eight newly-sequenced strains, with Arthrinium crenatum (CBS 146353) as the outgroup taxon. Multi-locus sequences contain 2,709 characters including gaps with ITS (1–610), LSU (611–1399), tef1 (1400–1948) and tub2 (1949–2691). Of these characters, 1,635 were constant, 367 were variable and parsimony-uninformative and 707 were parsimony-informative. For ML analysis, the matrix had 1,192 distinct alignment patterns. Estimated base frequencies were A = 0.229212, C = 0.248907, G = 0.263837, T = 0.258044; substitution rates: AC = 1.129211, AG = 2.936388, AT = 0.925501, CG = 0.917970, CT = 4.199729, GT = 1.0; gamma distribution shape parameter: α = 0.250690; and likelihood value of ln: -22 496.696950.

The ML tree topology agreed with the BI analysis and, therefore, only the ML tree is presented (Fig. 1). The strains obtained in this study were categorised into four clades, representing one known species and three new species (Fig. 1). The known species is A. arundinis and three new species are now recognised as A. coryli, A. lophatheri and A. oenotherae.

Figure 1. 

Phylogram of Apiospora, based on combined ITS, LSU, tef1 and tub2 genes. ML bootstrap support values (≥ 50%) and Bayesian posterior probability (≥ 0.90) are shown as first and second position above nodes, respectively. Strains from this study are shown in blue boxes, ex-type or ex-epitype cultures are indicated in bold face. Some branches were shortened according to the indicated mulipliers.

Taxonomy

Apiospora arundinis (Corda) Pintos & P. Alvarado, Fungal Systematics and Evolution 7: 205 (2021)

Fig. 2

Description

Asexual morph : Mycelium consisting of smooth, hyaline, branched, septate, 1.1–5.9 µm diam. hyphae (n = 20). Conidiophores reduced to conidiogenous cells. Conidiogenous cells subglobose to ampulliform, erect, blastic, aggregated in clusters on hyphae, smooth, branched, 3.4–9.4 × 1.5–6.4 µm, mean (± SD): 6.8 (± 1.6) × 3.9 (± 1.3) µm (n = 50). Conidia globose, subglobose to lenticular, with a longitudinal germ slit, occasionally elongated to ellipsoidal, brown to dark brown, smooth to finely roughened, 6.4–10.4 × 5.2–8.3 µm, mean (± SD): 7.7 (± 0.6) × 6.8 (± 0.7) µm, L/W = 1.0–1.5 (n = 50). Sexual morph: Undetermined.

Figure 2. 

Apiospora arundinis (CFCC 58977) A leaf of host plant B colony on PDA C conidiomata formed in culture D, E conidiogenous cells giving rise to conidia F conidia. Scale bars: 1000 µm (C); 10 µm (D–F).

Culture characteristics

On PDA, colonies thick and dense, margin undulate and irregular, pale yellow pigment diffused into medium, surface with patches of iron-grey aerial mycelia, reverse yellowish-brown, mycelia white to grey, sporulation on hyphae, reaching 9 cm in 7 days at 25 °C.

Specimens examined

China, Yunnan Province: Xishuangbanna Botanical Garden, on diseased leaves of Brunfelsia brasiliensis, 6 June 2022, S.J. Li, BJFC-S1918; living cultures CFCC 58977, LS 107).

Notes

In this study, two isolates clustered together with the culture of A. arundinis with high-support values (ML/BI = 100/0.99)in the multi-locus phylogenetic tree (Fig. 1). Thus, these isolates were identified as A. arundinis and Brunfelsia brasiliensis as a new host record for this species. Apiospora arundinis was introduced from Phyllostachys praecox, Castanea mollissima and Saccharum officinarum in China (Chen et al. 2014; Jiang et al. 2021; Liao et al. 2022). Comparing with the description from Chen et al. (2014) (5–7 × 2–4 µm), Jiang et al. (2021) (3–4 µm) and Liao et al. (2022) (4.5–7.4 × 3.3–4.4 µm), the conidia in this study show larger sizes (6.4–10.4 × 5.2–8.3 µm). These differences may result from different host and habitat.

Apiospora coryli S.J. Li & C.M. Tian, sp. nov.

MycoBank No: 849126
Fig. 3

Type

China, Shanxi Province: Ankang City, Huoditang Forest Farm, on dead plant culms of Corylus yunnanensis, 16 July 2021, R. Yuan & S.J. Li, holotype BJFC-S1920, ex-type living cultures CFCC 58978, CFCC 58979.

Etymology

Named after the host from which it was isolated.

Description

Asexual morph : Derived from sporulated cultures on PDA, hyphae hyaline, branched, septate, 1.1–5.2 µm diam. Conidiophores reduced to conidiogenous cells. Conidiogenous cells erect, aggregated in clusters on hyphae, hyaline to pale brown, smooth, doliiform to clavate or lageniform, 2.6–10.6 × 2.1–5.8 µm, mean (± SD): 5.5 (± 2.4) × 3.4 (± 1.1) µm (n = 50). Conidia brown to dark brown, globose to subglobose, oval or irregular, smooth to finely roughened, guttulate, usually with a longitudinal germ slit, 7.4–18.4 × 6.2–12.5 µm, mean (± SD): 10.8 (± 1.7) × 9.4 (± 1.3) µm, L/W = 0.8–1.6 (n = 50). Sexual morph: Undetermined.

Figure 3. 

Apiospora coryli (CFCC 58978, ex-holotype culture) A leaf of host plant B colony on PDA C conidiomata formed in culture D, E conidiogenous cells giving rise to conidia F conidia. Scale bars: 1000 µm (C); 10 µm (D–F).

Culture characteristics

On PDA, colonies circular, flat, entire margin, thick and cottony, concentrically spreading with aerial mycelium, margin regular, reddish-brown pigment diffused into medium, surface dark yellowish-brown, reverse dark reddish-brown to yellowish-brown from the centre, mycelia white to pale umber, sporulation on hyphae, reaching 9 cm in 7 days at 25 °C.

Notes

Strains of A. coryli constitutes a distinct clade, but there is poor support value in concatenated gene trees (Fig. 1). The most prominent distinguishing characteristic is the production of reddish-brown pigments on the culture medium.

Apiospora lophatheri S.J. Li & C.M. Tian, sp. nov.

MycoBank No: 849123
Fig. 4

Type

China, Yunnan Province, Xishuangbanna Primeval Forest Park, on diseased leaves of Lophatherum gracile, 4 June 2022, S.J. Li, holotype BJFC-S1917; ex-type living cultures CFCC 58975, CFCC 58976.

Etymology

Named after the host from which it was isolated.

Description

Asexual morph : Sporulated on PDA, mycelium consisting of hyaline, smooth, branched, septate hyphae 1.0–5.2 µm in diam. (n = 20). Conidiophores reduced to conidiogenous cells. Conidiogenous cells aggregated in clusters on hyphae, hyaline to pale brown, smooth, doliiform, clavate to ampulliform, 2.2–11.9 × 2.2–4.9 µm, mean (± SD): 6.4 (± 2.5) × 3.4 (± 0.6) µm (n = 50). Conidia globose, subglobose to lenticular, with a longitudinal germ slit, olive to dark brown, smooth to finely roughened and two or more conidia are produced on each conidiogenous cell, 5.1–8.9 × 4.6–7.7 µm, mean (± SD): 6.5 (± 0.8) × 5.9 (± 0.7) µm, L/W = 1.0–1.4 (n = 50). Sexual morph: Undetermined.

Figure 4. 

Apiospora lophatheri (CFCC 58975, ex-holotype culture) A leaf of host plant B colony on PDA C conidiomata formed in culture D conidiogenous cells giving rise to conidia E, F conidia. Scale bars: 1000 µm (C); 10 µm (D–F).

Culture characteristics

On PDA, colonies flat, spreading, margin circular, thick, concentrically spreading with aerial mycelium, surface light greyish-brown, reverse tawny pigment diffused in media, mycelia white to grey and pale brown, sporulation on hyphae, reaching 9 cm in 7 days at 25 °C.

Notes

Phylogenetic analysis indicated that Apiospora lophatheri is closely related to a clade comprising A. chromolaenae, A. euphorbiae, A. italicum, A. malaysiana, A. phyllostachydis, A. thailandica and A. vietnamense (Fig. 1). We compared the new species with phylogenetically similar taxa, based on morphological differences (Table 3) and base pair differences (Table 4). A. lophatheri can be differentiated from A. chromolaenae by its wider conidiogenous cells (2.2–11.9 × 2.2–4.9 µm vs. 6.5–12 × 1–2 µm) (from Euphorbia sp.; collected in Zambia; Ellis (1965)) and by 18 gene base pair differences (17/529 in ITS, 1/838 in LSU). A. lophatheri differs from A. euphorbiae by its larger olive to dark brown conidia (5.1–8.9 × 4.6–7.7 µm vs. 4–5.5 × 3–4 µm) (from Euphorbia sp.; collected in Zambia; Ellis (1965)), with nucleotide differences in ITS as 3/529, in LSU as 2/318, in tub2 as 22/801. A. italicum has smaller conidia (4–6 × 3–4 µm) (from Arundo donax; collected in Italy; Pintos et al. (2019)) and has 125 nucleotides differences (41/552 in ITS, 2/828 in LSU, 27/432 in tef1, 55/838 in tub2). Additionally, A. lophatheri is distinguished from A. malaysiana by having larger globose or subglobose conidia (5.1–8.9 × 4.6–7.7 µm vs. 5–6 × 3–4 µm) (from Macaranga hullettii; collected in Malaysia; Crous and Groenewald (2013)), with 43 nucleotide differences (3/529 in ITS, 1/838 in LSU, 18/424 in tef1, 21/801 in tub2). A. lophatheri differs from A. phyllostachydis by its relatively shorter conidiogenous cells (2.2–11.9 × 2.2–4.9 µm vs. 20–55 × 1.5–2.5 µm) (from Phyllostachys heteroclada; collected in China; Yang et al. (2019)) and by 48 nucleotides differences (7/529 in ITS, 3/838 in LSU, 12/424 in tef1, 26/795 in tub2). A. lophatheri can be differentiated from A. thailandica by having shorter conidiogenous cells (2.2–11.9 × 2.2–4.9 µm vs. 11.5–39 × 2–3.5 µm) (from bamboo; collected in Thailand; Dai et al. (2017)) and by 12 nucleotides differences (9/529 in ITS, 3/828 in LSU). The conidia of A. lophatheri are significantly wider and paler-coloured than those of A. vietnamense (5.1–8.9 × 4.6–7.7 µm vs. 5–6 × 3–4 µm) (from Citrus sinensis; collected in Vietnam; Wang et al. (2018)) and there are 7 nucleotides differences between the two species (2/526 in ITS, 2/803 in LSU, 3/315 in tub2). Therefore, A. lophatheri is described as a new species, based on phylogeny and morphological comparison.

Table 3.
Download as
CSV
XLSX

Summary of morphology of new Apiospora species and phylogenetic related species.

Species Isolation source Country Conidiogenous cells (µm) Conidia in surface view Conidia in side view References
Shape Diam (μm) Shape Diam (μm)
A. gaoyouense Phragmites australis China 1–2 × 2–3 globose to elongate ellipsoid 5–8 lenticular 4–8 Jiang et al. (2018)
A. hispanicum Maritime sand Spain – globose to ellipsoid 7.5–8.5 × 6–7.5 lenticular 6.5 Larrondo (1992)
A. locuta-pollinis Brassica campestris China 3–7.5 × 3–6 globose to elongate ellipsoid 8–15× 5–9.5 – – Zhao et al. (2018)
A. longistroma Bamboo Thailand – asexual morph: Undetermined – – – Dai et al. (2017)
A. marii Beach sand/ Poaceae Spain 5–10 × 3–4.5 globose to elongate ellipsoid 8–10(−13) lenticular (5–)6(−8) Crous and Groenewald (2013)
A. mediterranei Airborn spore/ grass Spain – lentiform 9–9.5 × 7.5–9 – – Larrondo (1992)
A. oenotherae Oenothera biennis China 2.0–14.2 × 1.1–4.9 globose, subglobose to lenticular 6.6–13.9 × 5.5–10.1 – – This study
A. piptatheri Piptatherum miliaceum Spain 6–27 × 2–5 globose to elongate ellips oid 6–8 × 3–5 lenticular 4.5–6 Pintos et al. (2019)
A. pseudomarii Aristolochia debilis China 8–13 × 2.5–5 subglobose to ellipsoid 6–9 × 4.5–6 – – Chen et al. (2021)
A. chromolaenae Chromolaena odorata Thailand 6.5–12 × 1–2 elongated, broadly fliform to ampulliform 4–6×4.5–6.5 – – Mapook et al. (2020)
A. euphorbiae Bambusa Bangladesh – circular or nearly circular (4–)4.7(–5.5) lenticular (3–)3.2(–4) Sharma et al. (2014)
A. italicum Arundo donax Italy (3–)4–7(–9) × (1.5–)2–3(–5) globose 4–6×3–4 lenticular – Pintos et al. (2019)
A. lophatheri Lophatherum gracile China 2.2–11.9 × 2.2–4.9 globose, subglobose to lenticular 5.1–8.9 × 4.6–7.7 – – This study
A. malaysiana Macaranga hullettii Malaysia 4–7 × 3–5 globose 5–6 lenticular 3–4 Crous and Groenewald (2013)
A. phyllostachydis Phyllostachys heteroclada China 20–55 × 1.5–2.5 globose to subglobose, oval or irregular 5–6 × 4–6 – – Yang et al. (2019)
A. thailandicum Bamboo Thailand 11.5–39 × 2–3.5 globose to subglobose, elongated to ellipsoidal 5–9 × 5–8 – – Dai et al. (2017)
A. vietnamense Citrus sinensis Vietnam 4–7 × 3–5 globose 5–6 lenticular 3–4 Wang et al. (2017)
Table 4.
Download as
CSV
XLSX

DNA base differences comparing Apiospora lophatheri sequences and sequences from related species.

Taxa Loci Nucleotides difference without gaps Rates of base pair differences
A. chromolaenae ITS 17/529 (40, 102, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122) 3.21%
LSU 1/838 (426) 0.12%
A. euphorbiae ITS 3/515 (26, 88, 89) 0.58%
LSU 2/318 (146, 306) 0.63%
tub2 22/801 (95, 96, 123, 151, 154, 163, 166, 182, 185, 193, 216, 237, 312, 347, 372, 429, 453, 454, 474, 559, 569, 574) 2.75%
A. italicum ITS 41/552 (40, 82, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 132, 165, 177, 180, 205, 207, 213, 487, 529) 7.43%
LSU 2/828 (406, 416) 0.24%
tef1 27/432 (16, 18, 19, 20, 21, 22, 23, 24, 25, 27, 35, 46, 53, 60, 75, 80, 90, 102, 119, 123, 125, 172, 210, 211, 240, 248, 272) 6.25%
tub2 55/838 (5, 29, 44, 45, 46, 92, 99, 119, 121, 122, 126, 155, 157, 171, 185, 188, 193, 194, 196, 198, 202, 297, 219, 229, 240, 265, 315, 338, 358, 363, 367, 368, 382, 384, 386, 390, 403, 407, 412, 430, 434, 454, 463, 465, 467, 480, 491, 499, 502, 556, 564, 580, 642, 756, 757) 6.56%
A. malaysiana ITS 3/529 (40, 102, 103) 0.57%
LSU 1/838 (426) 0.12%
tef1 18/424 (15, 16, 19, 27, 29, 38, 52, 56, 82, 83, 91, 93, 95, 111, 115, 202, 203, 264) 4.25%
tub2 21/801 (95, 96, 123, 151, 154, 163, 166, 182, 185, 193, 216, 237, 312, 347, 372, 429, 453, 474, 559, 569, 574) 2.62%
A. phyllostachydis ITS 7/529 (40, 44, 85, 102, 106, 433, 500) 1.32%
LSU 3/838 (7,8,9) 0.36%
tef1 12/424 (16, 19, 26, 27, 51, 52, 53, 111, 197, 202, 203, 264) 2.83%
tub2 26/795 (35, 52, 55, 84, 89, 112, 116, 147, 151, 175, 178, 186, 209, 211, 231, 329, 352, 354, 360, 462, 469, 489, 570, 572, 575, 608) 3.27%
A. thailandicum ITS 9/529 (40, 82, 102, 107, 122, 175, 177, 183, 501) 1.70%
LSU 3/828 (5, 416, 434) 0.36%
A. vietnamense ITS 2/526 (37, 99) 0.38%
LSU 2/803 (237, 391) 0.25%
tub2 3/315 (72, 82, 87) 0.95%

Apiospora oenotherae S.J. Li & C.M. Tian, sp. nov.

MycoBank No: 849125
Fig. 5

Type

China, Yunnan Province, Lincang City Triangle Plum Garden, on diseased leaves of Oenothera biennis, 26 April 2022, S.J. Li, holotype BJFC-S1919, ex-type living cultures CFCC 58972, LS 395.

Etymology

Named after the host from which it was isolated.

Description

Asexual morph : Hyphae hyaline, branched, septate, 1.2–4.8 µm in diam. (n = 20). Conidiophores reduced to conidiogenous cells. Conidiogenous cells smooth, ampulliform to doliiform, 2.0–14.2 × 1.1–4.9 µm, mean (± SD): 5.4 (± 2.9) × 3.1 (± 1.1) µm (n = 50). Conidia globose, subglobose to lenticular, with a longitudinal germ slit, occasionally elongated to ellipsoidal, colourless to dark brown, smooth to finely roughened, 6.6–13.9 × 5.5–10.1 µm, mean (± SD): 8.9 (± 1.2) × 7.8 (± 1.1) µm, L/W = 1.0–1.5 (n = 50). Sexual morph: Undetermined.

Figure 5. 

Apiospora oenotherae (CFCC 58972, ex-holotype culture) A leaf of host plant B colony on PDA C conidiomata formed in culture D, E conidiogenous cells giving rise to conidia F conidia. Scale bars: 1000 µm (C); 10 µm (D–F).

Culture characteristics

On PDA, colonies thick, concentrically spreading with aerial mycelium, circular, margin irregular, yellow to pale green pigment diffused into medium, surface with aerial mycelia, the reverse lightly pigmented with a few dark yellow patches, mycelia white to grey, sporulation occurs after 10 days, reaching 9 cm in 7 days at 25 °C.

Notes

Apiospora oenotherae belongs to the large clade, where it shows a relationship with A. gaoyouense, A. hispanicum, A. locuta-pollinis, A. longistroma, A. marii, A. mediterranei, A. piptatheri and A. pseudomarii (Fig. 1), but differs in distinct morphological characters (Table 3) and nucleotide differences (Table 5). A. oenotherae differs from A. gaoyouense by its production of significantly conidiogenous cells (2.0–14.2 × 1.1–4.9 µm vs. 1–2 × 2–3 μm) (from Phragmites australis; collected in China; Jiang et al. (2018)) and the presence of 30 distinct nucleotide positions (9/583 in ITS, 12/413 in tef1, 9/784 in tub2). A. oenotherae is distinct from A. hispanicum in producing larger conidial cells (6.6–13.9 × 5.5–10.1 µm vs. 7.5–8.5 × 6.2–7.6 µm) (from maritime sand; collected in Spain; Larrondo and Calvo (1992)) and in 30 nucleotides differences (1/539 in ITS, 1/320 in LSU, 28/796 in tub2). A. oenotherae differs from A. locuta-pollinis by its production of significantly conidiogenous cells (2.0–14.2 × 1.1–4.9 µm vs. 3–7.5 × 3–6 μm) (from hive-stored pollen; collected in China; Zhao et al. (2018)) and by the presence of 19 distinct nucleotide positions (1/539 in ITS, 7/416 in tef1, 11/485 in tub2). A. longistroma can be distinguished by growth rate, growing slowly on PDA, reaching 60 mm in 4 weeks (from bamboo; collected in Thailand; Dai et al. (2017)) and by the presence of 8 distinct nucleotide positions (6/572 in ITS, 2/840 in LSU). Moreover, A. mari produces elongated cells intermingled amongst conidia (from beach sand; collected in Spain; Crous and Groenewald (2013)), but A. oenotherae does not and can be distinguished by the presence of 23 distinct nucleotide positions (1/539 in ITS, 10/414 in tef1, 12/787 in tub2). Strains of A. mediterranei were isolated from pharmaceutical excipient, air-borne and on grass in Spain, while those of A. oenotherae collected from Oenothera biennis in China. There are no discernible morphological characters distinguishing these species, but the elongated stem branches and the presence of 30 distinct nucleotide positions (1/539 in ITS, 1/320 in LSU, 28/796 in tub2) serve as clear indicators of their distinct and phylogenetically well-separated taxa. A. oenotherae differs from A. piptatheri because of its wider conidial cells (6.6–13.9 × 5.5–10.1 µm vs. 6–8 × 3–5 μm) (from Piptatherum miliaceum; collected in Spain; Pintos et al. (2019)) and the presence of 14 distinct nucleotide positions (10/528 in ITS, 4/827 in LSU). It also differentiates from A. pseudomarii through the production of notably wider conidial cells (6.6–13.9 × 5.5–10.1 µm vs. 6–9 × 4.5–6 µm) and through 12 unique nucleotide positions (5/556 in tef1, 7/416 in tub2) (from Aristolochia debilis; collected in China; Chen et al. (2021)).

Table 5.
Download as
CSV
XLSX

DNA base differences comparing Apiospora oenotherae sequences and sequences from related species.

Taxa Loci Nucleotides difference without gaps Rates of base pair differences
A. gaoyouense ITS 9/583 (9, 10, 22, 36, 533, 535, 544, 555, 557) 1.54%
tef1 12/413 (34, 48, 56, 57, 69, 90, 122, 129, 134, 170, 226, 228) 2.91%
tub2 9/784 (538, 760, 766, 767, 768, 771, 775, 781, 782) 1.15%
A. hispanicum ITS 1/539 (528) 0.19%
LSU 1/320 (13) 0.31%
tub2 28/796 (30, 186, 539, 761, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 792, 794) 3.52%
A. locuta-pollinis ITS 1/539 (528) 0.19%
tef1 7/416 (33, 38, 94, 173, 177, 212, 258) 1.68%
tub2 11/485 (237, 459, 465, 466, 467, 470, 474, 480, 481, 483, 485) 2.27%
A. longistroma ITS 6/572 (20, 30, 38, 177, 213, 530) 1.05%
LSU 2/840 (655, 825) 0.24%
A. marii ITS 1/539 (528) 0.19%
tef1 10/414 (35, 49, 57, 58, 91, 123, 135, 171, 227, 229) 2.42%
tub2 12/787 (30, 186, 539, 761, 767, 768, 769, 772, 776, 782, 783, 785, 787) 1.52%
A. mediterranei ITS 1/539 (528) 0.19%
LSU 1/320 (13) 0.31%
tub2 28/796 (30, 186, 539, 761, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 792, 794) 3.52%
A. piptatheri ITS 10/528 (30, 38, 142, 177, 182, 213, 420, 421, 430, 431) 1.89%
LSU 4/827 (417, 431, 480, 632) 0.48%
A. pseudomarii ITS 5/556 (425, 528, 541, 560, 561) 0.90%
tef1 7/416 (33, 38, 94, 173, 177, 212, 258) 1.68%
tub2 1/718 (520) 0.14%

Discussion

Apiospora has been revised using different approaches and its taxonomy and classification have changed several times since its introduction. The taxonomic classification of the genus in relation to Arthrinium has been a topic of debate (Crous and Groenewald 2013; Pintos and Alvarado 2021). Morphologically, Apiospora and Arthrinium share similarities in basauxic conidiogenesis. The conidia of Apiospora are typically lenticular or obovoid in the side view, with colours ranging from pale brown to brown. Conversely, the conidia of Arthrinium exhibit various shapes, such as angular, curved, fusiform, globose, navicular and polygonal (Kunze 1817; Hyde et al. 1998; Wang et al. 2018; Pintos and Alvarado 2021).

Recently, several revisions have been made in the course of unitary nomenclature resulting in the discovery of a plethora of new species, based on multigene phylogenies (Kwon et al. 2021; Pintos and Alvarado 2021, 2022; Liu et al. 2023). Currently there are 93 accepted species in Apiospora (Table 2), which are found on a wide range of materials.

In this study, A. arundinis and A. lophatheri were collected from the tropical region of Xishuangbanna City, while A. coryli was discovered in Ankang City and A. oenotherae was found in Lincang City, which are both subtropical regions. Consistent with previous studies, the majority of Apiospora species inhabit a diverse range of habitats primarily located in tropical and subtropical regions (Pintos and Alvarado 2021).

Specimens of Apiospora were collected from the Qinling Mountains in Ankang City and, in addition to A. coryli, Jiang et al. reported species found including A. qinlingense and A. neochinensis (Jiang et al. 2018; Jiang et al. 2020). Amongst these species, A. coryli was found to have longer conidiogenous cells (2.6–10.6 × 2.1–5.8 µm) compared to A. qinlingense (1–2 × 2–3 µm) and A. neochinensis (1.5–6.5 × 1–3.5 µm) and much larger spores than A. qinlingense (4–18.4 × 6.2–12.5 µm vs. 5–8 × 5–8 µm) (Table 6). These morphological differences suggest that A. coryli is distinct from A. qinlingense and A. neochinensis. This distinction is also supported by phylogenetic analysis shown in Fig. 1 which revealed that these species are phylogenetically distant from each other. Different species have been discovered in this region over several years, indicating that variation in species may be linked to the timing of collection, host plants, growth rates, developmental cycles and activity levels. These findings highlight the diversity of fungi within Apiospora genus in the subtropical region of the Qinling Mountains and suggest the existence of numerous undiscovered species with significant research potential. Further investigation is necessary to determine the value of specific regions for future research on fungi.

Table 6.
Download as
CSV
XLSX

Synopsis of new Apiospora species and species collected from the Qinling Mountains in Apiospora.

Species Conidiogenous cells (µm) Conidia (µm) Host Date References
Apiospora coryli 2.6–10.6 × 2.1–5.8 4–18.4 × 6.2–12.5 Corylus yunnanensis 16 July 2021 Present study
A. qinlingense 1–2 5–8 Fargesia qinlingensis 27 June 2017 Jiang et al. 2018
A. neochinensis 1.5–6.5 × 1–3.5 8–12 × 5.5–9 Fargesia qinlingensis 16 July 2018 Jiang et al. 2020

* Newly described taxa are in bold.

This paper reports the initial discovery of A. lophatheri on Lophatherum gracile (Poaceae). While numerous Apiospora have been discovered on Poaceae plants worldwide, previous research has primarily focused on bamboo, with limited investigation into herbaceous plants, such as Lophatherum (Liu et al. 2023). However, prior to this study, Apiospora had not been previously found on Brunfelsia (Solanaceae) and Oenothera (Onagraceae). While Cercospora brunfelsiicola has been reported on other host Brunfelsia uniflora within the genus and Pestalotiopsis oenotberae has been identified specifically on Oenothera laciniata, the restricted cultivation of these plants along with insufficient research on their associated fungi have resulted in few related studies (Venkatasubbaiah et al. 1991; Hidayat and Meeboon 2014). This discovery highlights potential interactions between these plant species and their fungal counterparts, emphasising the importance of uncommon herbaceous plants for fungal taxonomy alongside Rosaceae and silvicultural species like Populus (Peng et al. 2022; Lin et al. 2022). Hence, collecting various specimens is crucial for studying and identifying the fungi of Apiospora, while also promoting fungal diversity.

Most Apiospora species exhibit round or lenticular conidia, as demonstrated in this study. Nevertheless, the sizes of these conidia often overlap amongst morphologically similar, but phylogenetically distinct species within the genus Apiospora. For example, the conidia of A. piptatheri (7.5–10 × 7–9 µm) and A. pseudosinense (8–10 × 7–10 µm) are similar, but the two species are comparable despite their distinct evolutionary lineages in Fig. 1 (Crous and Groenewald 2013; Pintos et al. 2019). Therefore, relying merely on morphology can pose challenges for accurate identification.

The monophyly of taxonomic classification units at every rank is crucially important. Morphology is frequently insufficient for phylogenetic classification and, thus, molecular evidence has become increasingly significant and indispensable for identifying and classifying fungal taxa. In recent years, there has been a steady growth in DNA sequencing data available for Apiospora species (Crous and Groenewald 2013; Wang et al. 2018; Pintos et al. 2019), leading to the recognition of 93 species of Apiospora. Sequence data are accessible for ITS in 93 species, LSU in 80, tef1 in 71 and tub2 in 73, facilitating accurate and swift identification (Wang et al. 2018; Pintos et al. 2019). However, using ITS alone has its limitations in identifying Apiospora species. Therefore, LSU, tef1, tub2 and multigene sequence data (ITS, LSU, tef1 and tub2) have been particularly useful in establishing phylogenetic relationships and increasing accuracy in Apiospora identification. Furthermore, this study yielded 32 sequence datasets for four gene regions (ITS, LSU, tef1 and tub2), enhancing our comprehension of the genus Apiospora. Novel species were identified by examining morphological and molecular characteristics, host associations and ecological distributions.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This study is financed by National Natural Science Foundation of China (Project No.: 31670647).

Author contributions

Conceptualization, Shuji Li and Chengming Tian; data curation, Shuji Li;funding acquisition, Chengming Tian; investigation, Shuji Li and Rong Yuan; project administration, Chengming Tian; resources, Shuji Li and Rong Yuan; supervision, Chengming Tian; writing-original draft, Shuji Li; writing-review and editing, Shuji Li, Cheng Peng, and Chengming Tian. All authors have read and agreed to the published version of the manuscript.

Author ORCIDs

Shuji Li https://orcid.org/0009-0006-4734-8399

Chengming Tian https://orcid.org/0000-0002-3352-7664

Data availability

All of the data that support the findings of this study are available in the main text or Supplementary Information.

References

  • Chen K, Wu XQ, Huang MX, Han YY (2014) First report of brown culm streak of Phyllostachys praecox caused by Arthrinium arundinis in Nanjing, China. Plant Disease 98(9): е1274. https://doi.org/10.1094/PDIS-02-14-0165-PDN
  • Chen TZ, Zhang Y, Ming XB, Zhang Q, Long H, Hyde KD, Li Y, Wang Y (2021) Morphological and phylogenetic resolution of Arthrinium from medicinal plants in Yunnan, including A. cordylines and A. pseudomarii spp. nov. Mycotaxon 136(1): 183–199. https://doi.org/10.5248/136.183
  • Cubeta MA, Echandi E, Abernethy T, Vilgalys R (1991) Characterization of anastomosis groups of binucleate Rhizoctonia species using restriction analysis of an amplified ribosomal RNA gene. Phytopathology 81(11): 1395–1400. https://doi.org/10.1094/Phyto-81-1395
  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12: 39–40.
  • Ellis MB (1965) Dematiaceous Hyphomycetes. VI. Mycological Papers 103: 1–46.
  • Feng Y, Liu JK, Lin CG, Chen YY, Xiang MM, Liu ZY (2021) Additions to the genus Arthrinium (Apiosporaceae) from bamboos in China. Frontiers in Microbiology 12: e661281. https://doi.org/10.3389/fmicb.2021.661281
  • 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. https://doi.org/10.1128/aem.61.4.1323-1330.1995
  • Hawksworth DL, Crous PW, Redhead SA, Reynolds DR, Samson RA, Seifert KA, Taylor JW, Wingfield MJ, Abaci Ö, Aime C, Asan A, Bai F-Y, de Beer ZW, Begerow D, Berikten D, Boekhout T, Buchanan PK, Burgess T, Buzina W, Cai L, Cannon PF, Crane JL, Damm U, Daniel H-M, van Diepeningen AD, Druzhinina I, Dyer PS, Eberhardt U, Fell JW, Frisvad JC, Geiser DM, Geml J, Glienke C, Gräfenhan T, Groenewald JZ, Groenewald M, de Gruyter J, Guého-Kellermann E, Guo LD, Hibbett DS, Hong SB, de Hoog GS, Houbraken J, Huhndorf SM, Hyde KD, Ismail A, Johnston PR, Kadaifciler DG, Kirk PM, Kõljalg U, Kurtzman CP, Lagneau PE, Lévesque CA, Liu X, Lombard L, Meyer W, Miller A, Minter DW, Najafzadeh MJ, Norvell L, Ozerskaya SM, Öziç R, Pennycook SR, Peterson SW, Pettersson OV, Quaedvlieg W, Robert VA, Ruibal C, Schnürer J, Schroers HJ, Shivas R, Slippers B, Spierenburg H, Takashima M, Taşkoin E, Thines M, Thrane U, Uztan AH, van Raak M, Varga J, Vasco A, Verkley G, Videira SIR, de Vries RP, Weir BS, Yilmaz N, Yurkov A, Zhang N (2011) The Amsterdam declaration on fungal nomenclature. IMA Fungus 2(1): 105–112. https://doi.org/10.5598/imafungus.2011.02.01.14
  • Hudson HJ (1960) Pyrenomycetes of sugar cane and other grasses in Jamaica. I. Conidia of Apiospora camptospora and Leptosphaeria sacchari. Transactions of the British Mycological Society 43(4): 607–616. https://doi.org/10.1016/S0007-1536(60)80051-4
  • Hyde KD, Fröhlich J, Taylor JE (1998) Fungi from palms. XXXVI. Reflections on unitunicate ascomycetes with apiospores. Sydowia 50: 21–80.
  • Jiang N, Fan XL, Tian CM (2021) Identification and characterization of leaf-inhabiting fungi from Castanea plantations in China. Journal of Fungi 7(1): 1–64. https://doi.org/10.3390/jof7010064
  • Katoh K, Rozewicki J, Yamada KD (2019) MAFFT online service: Multiple sequence alignment, interactive sequence choice and visualization. Briefings in Bioinformatics 20(4): 1160–1166. https://doi.org/10.1093/bib/bbx108
  • Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for bigger datasets. Molecular Biology and Evolution 33(7): 1870–1874. https://doi.org/10.1093/molbev/msw054
  • Kunze G (1817) Zehn neue Pilzgattungen. Mykol (1): 1–18.
  • Kwon SL, Park MS, Jang S, Lee YM, Heo YM, Hong JH, Lee H, Jang Y, Park JH, Kim C, Kim GH, Lim YW, Kim JJ (2021) The genus Arthrinium (Ascomycota, Sordariomycetes, Apiosporaceae) from marine habitats from Korea, with eight new species. IMA Fungus 12(1): 1–13. https://doi.org/10.1186/s43008-021-00065-z
  • Liao J, Jiang W, Wu X, He J, Li H, Wang T, Cheng L, Chen W, Mo L (2022) First report of Apiospora mold on Sugarcane in China caused by Apiospora arundinis (Arthrinium arundinis). Plant Disease 106(3): е1058. [Epub2022Feb16] https://doi.org/10.1094/PDIS-02-21-0386-PDN
  • Lin L, Pan M, Bezerra J, Tian CM, Fan XL (2022) Re-evaluation of the fungal diversity and pathogenicity of Cytospora species from Populus in China. Plant Disease 107(1): 183–196. https://doi.org/10.1094/PDIS-02-22-0260-RE
  • Liu RY, Li DH, Zhang ZX, Liu SB, Liu XY, Wang YX, Zhao H, Liu XY, Zhang XG, Xia JW, Wang YJ (2023) Morphological and phylogenetic analyses reveal two new species and a new record of Apiospora (Amphisphaeriales, Apiosporaceae) in China. MycoKeys 95: 27–45. https://doi.org/10.3897/mycokeys.95.96400
  • Mapook A, Hyde KD, McKenzie EHC, Bhat DJ, Jeewon R, Stadler M, Samarakoon MC, Malaithong M, Tanunchai B, Buscot F, Wubet T, Purahong W (2020) Taxonomic and phylogenetic contributions to fungi associated with the invasive weed Chromolaena odorata (Siam weed). Fungal Diversity 101(1): 1–175. https://doi.org/10.1007/s13225-020-00444-8
  • O’Donnell K, Kistler HC, Cigelnik E, Ploetz RC (1998) Multiple evolutionary origins of the fungus causing panama disease of banana: Concordant evidence from nuclear and mitochondrial gene genealogies. Proceedings of the National Academy of Sciences of the United States of America 95(5): 2044–2049. https://doi.org/10.1073/pnas.95.5.2044
  • Peng C, Crous PW, Jiang N, Fan XL, Liang YM, Tian CM (2022) Diversity of Sporocadaceae (pestalotioid fungi) from Rosa in China. Persoonia – Molecular Phylogeny and Evolution of Fungi 49: 201–260. https://doi.org/10.3767/persoonia.2022.49.07
  • Petrak F (1925) Mykologische Notizen VIII. Annales Mycologici 23: 1–143.
  • Pintos Á, Alvarado P (2022) New studies on Apiospora (Amphisphaeriales, Apiosporaceae): Epitypification of Sphaeria apiospora, proposal of Ap. marianiae sp. nov. and description of the asexual morph of Ap. sichuanensis. MycoKeys 92: 63–78. https://doi.org/10.3897/mycokeys.92.87593
  • Pintos Á, Alvarado P, Planas J, Jarling R (2019) Six new species of Arthrinium from Europe and notes about A. caricicola and other species found in Carex spp. hosts. MycoKeys 49: 15–48. https://doi.org/10.3897/mycokeys.49.32115
  • Réblová M, Miller AN, Rossman AY, Seifert KA, Crous PW, Hawksworth DL, Abdel-Wahab MA, Cannon PF, Daranagama DA, Wilhelm De Beer Z, Huang SK, Hyde KD, Jayawardena R, Jaklitsch W, Gareth Jones EB, Ju YM, Judith C, Maharachchikumbura SSN, Pang KL, Petrini LE, Raja HA, Romero AI, Shearer C, Senanayake IC, Voglmayr H, Weir BS, Wijayawarden NN (2016) Recommendations for competing sexual-asexually typified generic names in Sordariomycetes (except Diaporthales, Hypocreales, and Magnaporthales). IMA Fungus 7: 131–153. https://doi.org/10.5598/imafungus.2016.07.01.08
  • Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Hohna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61(3): 539–542. https://doi.org/10.1093/sysbio/sys029
  • Saccardo PA (1875) Conspectus generum pyrenomycetum italicorum additis speciebus fungorum Venetorum novis vel criticis, systemate carpologico dispositorum. Atti della Società Veneto-Trentina di Scienze Naturali 4: 77–100.
  • Samuels G, McKenzie E, Buchanan DE (1981) Ascomycetes of New Zealand 3. Two new species of Apiospora and their Arthrinium anamorphs on bamboo. New Zealand Journal of Botany 19(2): 137–149. https://doi.org/10.1080/0028825X.1981.10425113
  • Samarakoon MC, Hyde KD, Maharachchikumbura SSN, Stadler M, Jones EBG, Promputtha I, Suwannarach N, Camporesi E, Bulgakov TS, Liu JK (2022) Taxonomy, phylogeny, molecular dating and ancestral state reconstruction of Xylariomycetidae (Sordariomycetes). Fungal Diversity 112(1): 1–88. https://doi.org/10.1007/s13225-021-00495-5
  • Senanayake IC, Rossi W, Leonardi M, Weir A, McHugh M, Rajeshkumar KC, Verma RK, Karunarathna SC, Tibpromma S, Ashtekar N, Ashtamoorthy KS, Raveendran S, Kour G, Singh A, De la Peña-Lastra S, Mateos A, Kolařík M, Antonín V, Ševčíková H, Esteve-Raventós F, Larsson E, Pancorbo F, Moreno G, Altés A, Turégano Y, Du TY, Lu L, Li QR, Kang JC, Gunaseelan S, Kezo K, Kaliyaperumal M, Fu J, Samarakoon MC, Gafforov Y, Teshaboeva S, Kunjan PC, Chamaparambath A, Flakus A, Etayo J, Rodriguez-Flakus P, Zhurbenko MP, de Silva NI, Tennakoon DS, Latha KPD, Manimohan P, Raj KNA, Calabon MS, Ahmadpour A, Heidarian Z, Alavi Z, Alavi F, Ghosta Y, Azizi R, Luo M, Zhao MP, Kularathnage ND, Hua L, Yang YH, Liao CF, Zhao HJ, Lestari AS, Jayasiri SC, Yu FM, Lei L, Liu JW, Karimi O, Tang SM, Sun YR, Wang Y, Zeng M, Htet ZH, Linaldeddu BT, Alves A, Phillips AJL, Bregant C, Montecchio L, Kesel AD, Hustad VP, Miller AN, Fedosova AG, Kučera V, Raza M, Hussain M, Chen YP, Thiyagaraja V, Gomdola D, Rathnayaka AR, Dissanayake AJ, Suwannarach N, Hongsanan S, Maharachchikumbura SSN, Dissanayake LS, Wijayawardene NN, Phookamsak R, Lumyong S, Jones EBG, Yapa PN, Wanasinghe DN, Xie N, Doilom M, Manawasinghe IS, Liu JK, Zhao Q, Xu B, Hyde KD, Song J (2023) [in press] Fungal diversity notes 1611–1716: Taxonomic and phylogenetic contributions on fungal genera and species emphasis in south China. Fungal Diversity.
  • Sharma R, Kulkarni G, Sonawane MS, Shouche YS (2014) A new endophytic species of Arthrinium (Apiosporaceae) from Jatropha podagrica. Mycoscience 55(2): 118–123. https://doi.org/10.1016/j.myc.2013.06.004
  • von Höhnel F (1919) Fragmente zur Mykologie. XXII Mitteilungen, nr. 1092 bis 1153. Sitzungsberichte der Akademie der Wissenschaften in Wien 127(8–9): 549–634.
  • Yang CL, Xu XL, Dong W, Wanasinghe DN, Liu YG, Hyde KD (2019) Introducing Arthrinium phyllostachium sp. nov. (Apiosporaceae, Xylariales) on Phyllostachys heteroclada from Sichuan province, China. Phytotaxa 406(2): 91–110. https://doi.org/10.11646/phytotaxa.406.2.2
  • Zhao YZ, Zhang ZF, Cai L, Peng WJ, Liu F (2018) Four new filamentous fungal species from newly-collected and hive-stored bee pollen. Mycosphere: Journal of Fungal Biology 9(6): 1089–1116. https://doi.org/10.5943/mycosphere/9/6/3

Supplementary material

Supplementary material 1 

Isolates and GenBank accession numbers used in the phylogenetic analyses

Shuji Li, Cheng Peng, Rong Yuan, Chengming Tian

Data type: docx

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (38.08 kb)
login to comment