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Research Article
Morpho-phylogenetic evidence reveals new species in Rhytismataceae (Rhytismatales, Leotiomycetes, Ascomycota) from Guizhou Province, China
expand article infoJin-Feng Zhang§|, Jian-Kui Liu§, Kevin D. Hyde|#¤, Anusha H. Ekanayaka|¤, Zuo-Yi Liu§
‡ nstitute of Tea Research, Guizhou Academy of Agricultural Sciences, Guiyang, China
§ Guizhou Key Laboratory of Agriculture Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang, China
| Mae Fah Luang University, Muang, Thailand
¶ University of Electronic Science and Technology of China, Chengdu, China
# Zhongkai University of Agriculture and Engineering, Guangzhou, China
¤ Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
Open Access

Abstract

Karst formations represent a unique eco-environment. Research in the microfungi inhabiting this area is limited. During an ongoing survey of ascomycetous microfungi from karst terrains in Guizhou Province, China, we discovered four new species, which are introduced here as Hypoderma paralinderae, Terriera karsti, T. meitanensis and T. sigmoideospora placed in Rhytismataceae, based on phylogenetic analyses and morphological characters. Molecular analyses, based on concatenated LSU-ITS-mtSSU sequence data, were used to infer phylogenetic affinities. Detail descriptions and comprehensive illustrations of these new taxa are provided and relationships with the allied species are discussed, based on comparative morphology and molecular data.

Keywords

four new taxa, Hypoderma, karst formations, taxonomy, Terriera

Introduction

Rhytismataceae (Rhytismatales) was established by Chevallier (1826), typified by Rhytisma with R. acerinum (Pers.) Fr. as the type species and belongs in Rhytismatales, Leotiomycetes, Ascomycota (Wijayawardene et al. 2020). Members of this family produce variously shaped apothecia that may be sessile, circular, navicular or hysteriform and that typically open by a longitudinal split or radial fissures. Asci are cylindrical, saccate to clavate. Ascospores are one-celled or multi-septate and vary from bacilliform to fusiform or filiform, with or without a sheath (Darker 1967; Ekanayaka et al. 2019). Species of Rhytismataceae occur on a wide range of hosts with a worldwide distribution (Cannon and Minter 1986; Johnston 1986; Hou and Piepenbring 2009; Hernández et al. 2014; Li et al. 2014; Tanney and Seifert 2017; Cai et al. 2020).

Darker (1967) proposed the generic delimitation for Rhytismataceae, based on ascoma and ascospore shapes, although this has been challenged in later studies (Cannon and Minter 1986; Johnston 1990, 2001; Hou et al. 2005). However, Darker (1967) and Cannon and Minter (1986) were followed due to lack of an alternative scheme. Molecular studies (Gernandt et al. 2001; Johnston and Park 2007; Lantz et al. 2011; Tian et al. 2013; Zhang et al. 2015) had revealed the phylogenetic relationships amongst members of Rhytismatales, but the available sequence data for this group remains limited and a phylogenetic classification of some members is unresolved. There are around 50 genera with 1000 species presently accepted in Rhytismataceae (Lumbsch and Huhndorf 2007; Wijayawardene et al. 2018; Index Fungorum 2020); however, a systematic genus-level taxonomic revision is needed to provide a clear, natural generic delimitation within this family and the relationship between Rhytismataceae and allied families within Rhytismatales needs to be resolved (Johnston et al. 2019).

Karst formations are generally characterised by sinking streams, caves, enclosed depressions, fluted rock outcrops and large springs (Ford and Williams 2007). Guizhou, as the eastern portion of the Yunnan-Guizhou Plateau, has the largest proportion of rocky desertification and karst landforms in China (Huang and Cai 2006). The flora in this area, comprising of 264 families with 1667 genera and 7505 vascular plants species, were inventoried from Guizhou Province (Liu et al. 2018). Therefore, it would be interesting to study the fungi in this area because of its unique ecological environment and rich plant resources. A series of studies have already been carried out and yielded several new species (Zhang et al. 2016, 2017a, b, 2018, 2019). The objectives of this study are to introduce four novel species of Rhytismataceae, based on phylogenetic and morphological evidence and elucidate their affinities with related species.

Materials and methods

Collection, examination, isolation and specimen deposition

Specimens were collected from Guizhou Province from 2016 to 2017 and examined in the laboratory with a Motic SMZ 168 stereomicroscope. Vertical sections of fruiting bodies were made by hand and mounted in water for microscopy. Macro-morphological characters were captured using a stereomicroscope (Nikon SMZ800N) with a Cannon EOS 70D digital camera. Micro-morphological characters were observed by differential interference contrast (DIC) using a Nikon ECLIPSE 80i compound microscope and captured by a Cannon EOS 600D digital camera. Measurements were processed in a Tarosoft (R) Image Frame Work version 0.9.7 programme and photographic plates were edited in Adobe Photoshop CS6 (Adobe Systems Inc., USA).

The single spore isolation technique described in Chomnunti et al. (2014) was followed to obtain the pure cultures of these specimens. Single germinated ascospore was picked up and transferred to potato dextrose agar (PDA; 39 g/l distilled water, Difco potato dextrose) for recording growth rates and culture characteristics.

The holotypes are deposited at the Herbarium of Mae Fah Luang University (MFLU), Chiang Rai, Thailand or Guizhou Academy of Agricultural Sciences (GZAAS), Guizhou, China. Ex-type living culture is deposited at Guizhou Culture Collection (GZCC), Guiyang, China. Index Fungorum and Facesoffungi numbers are provided according to Jayasiri et al. (2015) and Index Fungorum (2020). New species were established, based on the recommendations from Jeewon and Hyde (2016).

DNA extraction, PCR and phylogenetic analyses

Following the manufacturer’s instructions, the total genomic DNA was extracted from cultures using a Biospin Fungus Genomic DNA Extraction Kit (BioFlux, Hangzhou, P. R. China) or extracted from the fruiting bodies using an E.Z.N.A. Forensic DNA kit (Omega Bio-Tek, Doraville, Georgia, USA).

Polymerase chain reactions (PCR) were performed in 25 μl reaction volumes, which contained 9.5 μl distilled-deionised-water, 12.5 μl of 2 × Power Taq PCR Master Mix (TIANGEN Co., China), 1 μl of DNA template and 1 μl of each forward and reverse primers. Three different loci were used in this study. The internal transcribed spacer (ITS) and 28S large subunit of the nuclear ribosomal DNA (LSU) regions were amplified by using the primers ITS4/ITS5 and LR0R/LR5, respectively (White et al. 1990; Gardes and Bruns 1993). The primers mrSSU1 and mrSSU3R were used for amplification of the mitochondrial small subunit (mtSSU) partial regions (Zoller et al. 1999). The PCR thermal cycle programme was performed according to White et al. (1990), Gardes and Bruns (1993) and Zoller et al. (1999). Amplicon size and concentration were assessed by gel electrophoresis with 1.2% agarose stained with ethidium bromide. PCR products were purified and sequenced at Sangon Biotechnology Co. Ltd (Shanghai, P. R. China).

For phylogenetic reconstruction, newly-generated sequences were initially subjected to BLAST search (BLASTn) in NCBI (https://www.ncbi.nlm.nih.gov) and additional related sequences were selected and downloaded from GenBank (https://www.ncbi.nlm.nih.gov/genbank/), based on BLASTn results and recent publications (Tian et al. 2013; Wang et al. 2013; Zhang et al. 2015; Johnston et al. 2019; Cai et al. 2020). The sequences used in this study for phylogenetic analysis are listed in Table 1. All of these sequences were aligned and manually improved with BioEdit v. 7.2 (Hall 1999) and then assembled as a dataset of LSU-ITS-mtSSU to infer the phylogenetic placement of newly identified taxa.

Table 1.

Taxa used in this study. Strains generated/sequenced in this study are given in bold.

Taxa Specimen/Strain No. GenBank accession numbers
LSU ITS mtSSU
Bifusella camelliae HOU 1094 KF797447 KF797435 KF797458
HOU 701B KF797448 KF797436 KF797459
Coccomyces anhuiensis BJTC 201610 MK371314 MK371313 MK371315
Coccomyces dentatus AFTOL ID-147 AY544657 DQ491499 AY544736
Colpoma ledi Lantz 379 (UPS) HM140512 HM143788
Colpoma quercinum Lantz 368 (UPS) HM140513 HM143789
Cryptomyces maximus Lantz and Minter 424 (UPS) HM140514 HM143790
Discocainia nervalis BITC 201405 KJ513473 KJ507206
Duplicariella phyllodoces Lantz 389 (UPS) HM140516
Hypoderma berberidis HOU 892 JX232420 JX232414 KF813010
HOU 942 JX232421 JX232415 KF813009
Hypoderma campanulatum ICMP 17383 HM140517 HM143792
Hypoderma carinatum ICMP 18322 HM140518 HM143793
Hypoderma cordylines ICMP 17344 HM140521 JF683421 HM143796
ICMP 17396 HM140520 HM143795
Hypoderma hederae Lantz and Minter 421 (UPS) HM140522 JF690770 HM143797
Hypoderma liliense ICMP 18323 HM140523 MH921859 HM143798
ICMP 18324 HM140524 HM143799
Hypoderma minteri BJTC 201203 JX232418 JX232416
Hypoderma obtectum ICMP 17365 HM140525 HM143800
Hypoderma paralinderae GZAAS 19-1769 MN638878 MN638873 MN638868
Hypoderma rubi Hanson 2006-451 (UPS) HM140519 JF690769 HM143794
ICMP 17339 HM140526 JF683419 HM143801
ICMP 18325 HM140527 JF683418 HM143802
Lantz 405 (UPS) HM140530 JF690772 HM143805
Hypoderma sticheri ICMP 17353 HM140529 MK039702 HM143804
Hypohelion anhuiense BITC 201311 KF797443 KF797431 KF797455
Hypohelion scirpinum Lantz 394 (UPS) HM140531 HM143806
Lirula macrospora Hou et al. 13 (BJTC) HQ902159 HQ902152
Lirula yunnanensis BJTC 2012 HQ902149 HQ902156
Lophodermium arundinaceum Lantz 323 (UPS) HM140535 HM143811
Lophodermium culmigenum ICMP 18328 HM140538 HM143814
Marthamyces emarginatus ICMP 22854 MK599203 MH921869 MK598751
Meloderma dracophylli ICMP 17343 HM140561 MH921871 HM143833
Nematococcomyces oberwinkleri BJTC 201205 KC312686 KC312689
Nematococcomyces rhododendri HOU 469A KC312687 KU213975 KC312691
Rhytisma huangshanense HOU 564 FJ495192 GQ253101
Rhytisma salicinum Lantz 370 (UPS) HM140566
Sporomega degenerans Lantz 367 (UPS) HM140567 HM143839
Terriera camelliicola AAUF 66555 KP878552 KP878553
Terriera cladophila Lantz & Minter 423 (UPS) HM140568 HM143840
Terriera elliptica BJTC 201419 KP878550 KP878549 KP878551
Terriera guihzouensis BITC 2020149 MT549890 MT534526
BITC 2020147 MT534519 MT549863
BITC 2020148 MT534527 MT549874
BITC 2020149 MT549872 MT534528 MT549865
BITC 2020150 MT534591 MT549888
Terriera houjiazhuangensis BITC 2020145 MT549889 MT549882
BITC 2020146 MT549864 MT549879 MT549884
BITC 2020192 MT549869 MT549883
Terriera ilicis BJTC 2020141 MT549885 MT549875 MT549868
BJTC 2020193 MT549873 MT549861 MT549886
BJTC 2020142 MT549881 MT549877 MT549870
Terriera karsti MFLU 18-2288 MN638881 MN638876 MN638871
Terriera meitanensis MFLU 18-2299 MN638879 MN638874 MN638869
Terriera meitanensis MFLU 18-2301 MN638880 MN638875 MN638870
Terriera minor ICMP 13973 HM140570 HM143842
Terriera pandanicola MFLU 16-1931 MH260320 MH275086 MW334971
Terriera sigmoideospora MFLU 18-2297 MN638882 MN638877 MN638872
Terriera thailandica MFLUCC 14-0818 KX765301
Therrya abieticola HOU 447A KP322580 KP322574 KP322587
Tryblidiopsis pinastri CBS 445.71 MH871979 JF793678 AF431963
Tryblidiopsis sichuanensis BJTC 201211 KC312683 KC312676 KC312692
Tryblidiopsis sinensis BJTC 201212 KC312681 KC312674 KC312694

Phylogenetic analyses were performed using the algorithm of Maximum-Parsimony (MP) and Bayesian Inference (BI). MP analyses were run using PAUP v. 4.0b10 (Swofford 2002) with 1000 replications and inferred using the heuristic search option with 1000 random taxa. All characters were unordered and of equal weight and gaps were treated as missing data. Maxtrees was set as 1000, zero-length branches were collapsed and all equally parsimonious trees were saved. Clade stability was accessed using a bootstrap (BT) analysis with 1000 replicates, each with ten replicates of random stepwise addition of taxa (Hillis and Bull 1993).

BI analyses were carried out by using MrBayes v. 3.2 (Ronquist et al. 2012). The best-fit model (GTR+I+G for LSU, ITS and mtSSU) of evolution was estimated in MrModeltest 2.3 (Nylander 2008). Posterior Probabilities (PP) (Rannala and Yang 1996; Zhaxybayeva and Gogarten 2002) were determined by Markov Chain Monte Carlo sampling (MCMC) in MrBayes v. 3.2. Six simultaneous Markov chains were run for 10,000,000 generations and trees were sampled every 100th generation. The temperature values were lowered to 0.15, burn-in was set to 0.25 and the run was automatically stopped as soon as the average standard deviation of split frequencies reached below 0.01.

The phylogram was visualised in TreeView (Page 1996) and edited in Adobe Illustrator CS v. 5 (Adobe Systems Inc., USA). The finalised alignment and tree were deposited in TreeBASE, submission ID: 27401 (http://www.treebase.org).

Results

Phylogenetic analyses

The dataset for phylogenetic analysis comprised 64 strains, with Marthamyces emarginatus (Cooke & Massee) Minter selected as the outgroup taxon. This dataset consists of 2078 characters (including the gaps), of which 1205 are constant, 236 are variable parsimony-uninformative, while 637 characters are parsimony-informative. The most parsimonious tree showed with length of 2843 steps (CI = 0.480, RI = 0.759, RC = 0.364 and HI = 0.520). The best tree revealed by the MP analysis was selected to represent relationships amongst taxa (Fig. 1). The tree generated from Bayesian inference analyses had similar topology. The phylogram (Fig. 1) shows that Hypoderma is non-monophyletic (Clade A, B, C and D), with H. paralinderae clusters with three existing species viz. H. cordylines P.R. Johnst., H. hederae (T. Nees ex Mart.) De Not. and H. rubi (Pers.) DC. In contrast, all of the Terriera species with available sequences (including the newly generated sequences) form a monophyletic clade with strong statistical support (MPBP 100% and BYPP 1.00). This corresponds to the phylogeny in Zhang et al. (2015). Terriera meitanensis and T. karsti group together with three reported species viz. T. camelliicola (Minter) Y.R. Lin & C.L. Hou, T. elliptica T.T. Zhang & C.L. Hou and T. thailandica Jayasiri & K.D. Hyde, while T. sigmoideospora is placed within another clade that comprises T. houjiazhuangensis C.L. Hou & S.R. Cai and T. pandanicola Tibpromma & K.D. Hyde.

Figure 1. 

Phylogram of Rhytismataceae is presented as the best tree revealed by MP analysis, based on the concatenated LSU-ITS-mtSSU sequence dataset. MP bootstrap support values (MPBP ≥ 50%) and Bayesian inference posterior probabilities (BYPP ≥ 0.95) are shown near the nodes. The tree is rooted to Marthamyces emarginatus (ICMP 22854), the scale bar showing 10 changes. Type strains are indicated in bold and new sequences, generated in this study, are given in red.

Taxonomy

Hypoderma De Not., G. bot. ital. 2(2): 13 (1847)

De Candolle (1805) introduced Hypoderma to accommodate taxa resembling Hysterium Pers., but with apothecia that are immersed in host-plant tissue and the hymenia are exposed via a longitudinal split in the substratum. Subsequently, the nomenclature of Hypoderma was challenged by various authors (Chevallier 1822, 1826; Fries 1823; Wallroth 1833). De Notaris (1847) recognised the distinction between Hypoderma and Lophodermium Chevall. and separated them, based on the ascospore shapes. So far, there are 214 epithets included in Index Fungorum (2020), but around half of these species are synonymized under other genera, such as Lophodermium, Meloderma Darker and Terriera.

Hypoderma paralinderae J.F. Zhang & Z.Y. Liu, sp. nov.

Figure 2

Etymology

Referring to the morphological similarity with Hypoderma linderae.

Holotype

GZAAS 19-1769.

Description

Apothecia developing on dead stems, scattered, dark brown to black, shiny, long elliptical to slightly fusiform, straight or somewhat curved, ends rounded or obtuse, rising above the surface of the substrate, opening by a single longitudinal split. Lips moderately developed, pale brown (Fig. 2a, b). In median vertical section (Fig. 2c), apothecia subcuticular, 200–280 µm deep. Covering stroma (Fig. 2e) up to 38–45 µm thick near the opening, becoming to 12–18 µm thick towards the edges, extending to the basal stroma, consisting of an outer layer of host cuticle and several layers of dark brown, thick-walled cells of textura angularis. Lip cells (Fig. 2d) clavate to cylindrical, 11–23 × 2–3 µm, thin-walled, hyaline to pale brown, 0–1-septate. Basal stroma (Fig. 2f) 10–16 µm thick, consisting of several layers of brown, thick-walled cells, arranged in textura angularis, becoming colourless, thin-walled cells of textura prismatica towards the subhymenium. Subhymenium 19–27 µm thick, composed of several layers of hyaline, thin-walled cells of textura angularis. Paraphyses 1.5–2 µm, filiform, aseptate, unbranched, often curved, but not swollen at the apex, anastomosing at the base. Asci (81.5–)110–120(–129) × 10–14 µm (x¯ = 108 × 12 µm, n = 25), 8-spored, unitunicate, cylindrical-clavate, round to subtruncate at the apex, with a 38–49 µm long stalk, thin-walled, J-, apical ring, without circumapical thickening. Ascospores 26–32.5 × 2.5–4.5 µm (x¯ = 30.5 × 3.5 µm, n = 35, measured without the gelatinous sheath), multi-seriate and mostly arranged in the upper half of ascus, fusiform to slightly cylindrical, straight or lightly curved, apex rounded and tapering slightly to an acute base, aseptate, hyaline, guttulate, surrounded by a 0.5–1.5 µm thick gelatinous sheath (extending to 2.5 µm at the poles). Asexual morph: Not observed.

Figure 2. 

Hypoderma paralinderae a, b apothecia observed under a dissecting microscope in face view c vertical section through an apothecium d lips adjacent to the top of covering stroma e section of covering stroma f section of basal stroma g paraphyses and asci in various states of maturity h immature ascus i, j ascospores. Note: c–j mounted in water. Scale bar: 1 mm (a), 500 µm (b), 200 µm (c), 20 µm (d, g, h), 10 µm (e, i, j), 5 µm (f).

Material examined

CHINA, Guizhou Province, Leishan County, dead stems of unidentified herbaceous plants, 2 November 2017, J.F. Zhang, LS-21 (GZAAS 19-1769, holotype).

Notes

Our phylogenetic analysis shows that Hypoderma paralinderae is placed in Hypoderma D clade (Fig. 1) and clustered with H. cordylines, H. hederae and H. rubi. Both H. paralinderae and H. codylines have similar sized asci (110–122.5 × 5.5–7 µm vs. 90–140 × 11–16 µm); however, they can be distinguished by the different shape and size of ascospores (fusiform to slightly cylindrical, 26–32.5 × 2.5–4.5 µm in H. paralinderae vs. elliptic, 14–21 × 4.5–6 µm in H. cordylines) (Johnston 1990). Hypoderma paralinderae shares similar-sized asci with H. hederae; however, it is differentiated from the latter by larger ascospores (26–32.5 × 2.5–4.5 µm vs. 18–22 × 3.5–4 µm) (Powell 1974). Moreover, H. hederae was described with oblong-cylindrical ascospores that are bluntly round on both ends; however, the ascospores in H. paralinderae are fusiform to cylindrical, but rounded at the apex and tapering slightly to an acute base (Powell 1974), while H. paralinderae differs from H. rubi by having obviously larger asci (110–122.5 × 5.5–7 µm vs. 60–100 × 10–12.5 µm) and ascospores (26–32.5 × 2.5–4.5 µm vs. 14–18 × 3.5–4.5 µm) (Hou et al. 2007). Besides, the recommendations of delineation taxa from Jeewon and Hyde (2016) are followed and comparisons of the ITS gene region between H. paralinderae and H. cordylines (ICMP 17344), as well as H. paralinderae and H. rubi (ICMP 17339) are processed. The results showed that there are 9/468 bp (1.9%) and 9/467 (1.9%) bp differences (including gaps) between them, respectively. According to the above evidence, H. paralinderae is introduced herein as new to science.

Terriera B. Erikss., Symb. bot. upsal. 19(no. 4): 58 (1970)

Terriera was segregated from Lophodermium by Eriksson (1970) with T. cladophila as its type species. Johnston (2001) elucidated some distinctive morphological features (described as oblong to sublinear ascomata with single longitudinal opening slit, narrow-cylindrical asci and 1-septate ascospores that taper slightly at both ends and often becoming gently sigmoid on release and lacking a gelatinous sheath) for this genus and justified its monophyletic classification. There are 38 species accepted in Terriera (Index Fungorum 2020) and around half of these species were discovered recently from China (Chen et al. 2011, 2013; Yang et al. 2011; Zheng et al. 2011; Gao et al. 2012; Song et al. 2012; Zhou et al. 2012; Li et al. 2015a, b; Lu et al. 2015; Wu et al. 2015; Cai et al. 2020). Here, we introduce three novel species. These three species share morphological characters typical of Terriera and cluster together with existing Terriera species in LSU-ITS-mtSSU phylogenetic analyses. In addition, a synopsis for Terriera species is also provided and listed in Table 2.

Table 2.

Synopsis of Terriera species. The new species described in this study are indicated in bold.

Species Host Appearance of apothecia Asci Ascospores Origin References
Terriera aequabilis On dead leaves of Photinia villosa Elliptical to sub-circular, straight or slightly curved to one side, ends rounded and opening by a single longitudinal split 75–105 × 4.5–5.5 µm 55–78 × 0.8–1 µm, filiform, aseptate, ends rounded, covered by a 0.3–0.5 µm wide gelatinous sheath Jiangxi, China Li et al. 2015b
T. angularis On leaves of Illicium simonsii Triangular to quadrangular, rarely elliptical and opening by 3–4 radial splits or a longitudinal split 105–130 × 5.5–6.5 µm 70–90 × 1–1.2 µm, filiform, aseptate, slightly tapering towards the round base, covered by a 0.8–1 µm wide gelatinous sheath Hubei, China Zhou et al. 2013
T. arundinacea On decomposed leaves of Bambusa sp. Oblong to sublinear and opening by a single longitudinal split 130–160 × 8–9 µm 90–100 × 2–2.5 µm, slightly tapering towards the base, lacking gelatinous sheath Java, Indonesia Johnston 2001
T. asteliae On dead leaves of Asterlia sp. Elliptical to oblong, ends rounded, opening by a single longitudinal slit 75–105 × 8–10.5 µm 45–70 × 2–2.5 µm, slightly tapering towards both ends and slightly constricted near the centre, aseptate or 1-septate, gently curved, lacking gelatinous sheath Northland, New Zealand Johnston 2001
T. breve On dead leaves of Carex, Unicinia and Gahnia spp. Oblong-elliptical, ends rounded, often sublinear, with a single longitudinal opening slit 110–135(–160) × 6–7 µm (55–)60–75 × 1.5–2 µm, slightly tapering towards both ends, aseptate or 1-septate, gently curved or sigmoid, lacking gelatinous sheath Campbell I, New Zealand Johnston 2001
T. camelliae On fallen leaves of Camellia sp. Subcircular to irregular bleached spots, elliptical or occasionally 3-lobed and opening by a longitudinal split 85–120 × 5.5–6.5 µm 52–80 × 1–1.2 µm, filiform, aseptate, covered by a ca. 0.5 µm wide gelatinous sheath. Fuzhou, China Chen et al. 2011
T. camelliicola On twigs of Camellia sinensis Elliptical, occasionally fusing to form elongated elliptical, opening by a single longitudinal split 80–110 × 5–7 µm 50–70 × 1 µm, filiform, aseptate, covered by a 0.5 µm wide gelatinous sheath. Assam, India Minter and Sharma 1982
T. cladophila On dead twigs of Vaccinium myrtillus Elliptical, rounded at the ends, with a longitudinal opening split 75–100 × 5.5–8 µm 60–70 × 1 µm, filiform, aseptate, lacking gelatinous sheath Norway Terrier 1942; Eriksson 1970
T. clithris On dead leaves of unidentified monocotyledon Cylindrical to linear, with longitudinal opening slit 110–120 × 6.5–7.0 µm 60–80 × 1–1.5 µm, slightly tapering towards both ends, lacking gelatinous sheath Rio Grande Do Sul, Brazil Johnston 2001
T. coacervata On leaves of Lithocarpus cleistocarpus Elliptical, sometimes branching into lobed or polygonal shapes, opening by a longitudinal split or by more than 3 lobes 90–130 × 6.0–7.0 µm 60–110 × 1.5–1.8 µm, filiform, aseptate, covered by a 1.0–1.5 µm wide gelatinous sheath Anhui, China Zheng et al. 2012
T. dracaenae On dead leaves or stems of Dracaena sp. Oblong to oblong-elliptical, ends rounded, opening by a single longitudinal split 130–140 (–160) × 6–7 µm 100 × 2 µm, 1-septate, lacking gelatinous sheath California, USA Johnston 2001
T. elliptica On living twigs of Rhododendron sp. Elliptical, ends rounded to subacute, opening by a longitudinal split 135–175 × 7–9 μm 60–85 × 1.5–2 μm, filiform, slightly tapering towards both ends, aseptate, covered by a 1–1.5 μm wide gelatinous sheath Yunnan, China Zhang et al. 2015
T. fici On dead leaves of Ficus vasculosa Rounded or subrounded, with conspicuous edge and opening by a single longitudinal split 90–115 × 4–5.5 µm 65–80 × 0.8–1 µm, filiform, aseptate, rounded to obtuse at the apex, slightly tapering towards the rounded or subacute base, covered by a 0.5 µm wide gelatinous sheath Hainan, China Wu et al. 2016
T. fuegiana On dead leaves of Rostkovia grandiflora Oblong elliptical to broad-elliptical, ends rounded, opening by a single, longitudinal slit 75–95 × 7–10 μm 60–65 × 1.5–2.5 μm, slightly tapering towards both ends, 1-septate, lacking gelatinous sheath Tierra del Fuego, Argentina Johnston 2001
T. fourcroyae On dead leaves of Furcraea sp. Oblong-elliptical, ends rounded, with a single longitudinal opening slit 95–110 × 5–6.5 µm 60–70 × 1.5–2.5 μm, slightly tapering towards both ends, gently coiled or sigmoid, 1-septate, lacking gelatinous sheath. Sri Lanka Johnston 2001
T. guizhouensis On dead leaves of Eriobotrya japonica Elliptical, occasionally curved, opening by a longitudinal split 88–107 × 4–6 µm 50–80 × 1–1.2 µm, filiform, slightly tapering towards both ends, aseptate, pluriguttulate, covered by a thin gelatinous sheath Guizhou, China Cai et al. 2020
T. houjiashanensis On dead leaves of Ilex cornuta Elliptical, often curved, occasionally confluent, opening by a longitudinal split 103–128 × 4–6 µm 73–82 × 0.6–0.9 µm, filiform, slightly tapering towards both ends, aseptate, pluriguttulate, covered by an inconspicuous gelatinous sheath Anhui, China Cai et al. 2020
T. huangshanensis On leaves of Eurya muricata var. huiana Elliptical, fusiform or subfusiform, straight or curved (lunate), sometimes 3-lobed or triangular, ends rounded to subacute, opening by a single longitudinal split 100–120 × 5–7 µm 58–90 × 1.5–2 µm, filiform, slightly tapering towards the base, aseptate, covered by a 1–1.5 µm thick gelatinous sheath Anhui, China Yang et al. 2011
T. ilicis On dead leaves of Ilex pernyi Elliptical, occasionally curved, triangular or confluent, opening by a longitudinal split 117–139 × 4–7 µm 52–84 × ca. 1 µm, filiform, slightly tapering towards both ends, aseptate, pluriguttulate, covered by a thin gelatinous sheath Hubei, China Cai et al. 2020
T. illiciicola On dead leaves of Lithocarpus cleistocarpus Subcircular to broad-elliptical, opening by a longitudinal split 90–135 × 4.0–5.0 µm 65–95 × 1 µm, filiform, aseptate, covered by an inconspicuous gelatinous sheath Anhui, China Zheng et al. 2011
T. intraepidermalis On fallen leaves of Photinia prunifolia Widely elliptical, sometimes elliptical or subcircular, occasionally triangular, straight or curved to one side slightly, ends round to obtuse, opening by a single longitudinal split or by three radial splits 90–135 × 5.5–7.5 µm 70–105 × 1–1.5 µm, with upper end rounded to obtuse, slightly tapering towards the rounded base, covered by a 0.5 μm wide gelatinous sheath Hunan, China Lu et al. 2015
T. javanica On dead leaves of Elettaria sp. Oblong-elliptical to sublinear, ends acute, opening by a single longitudinal slit 85–95 × 5.5–7 µm 50–60 × 1.5 µm, but the detailed morphological characters were not seen Java, Indonesia Johnston 2001
T. karsti On dead branch of unidentified host Elliptical or oblong-elliptical, ends slightly acute to obtuse, with a single longitudinal opening split (103–)110–122.5 × 5.5–7 µm 55–66 × 1.5–2.0 µm, filiform, gradually tapering towards both ends, aseptate, lacking gelatinous sheath Guihzou, China In this study
T. latiascus On dead leaves of Euterpe and Heliconia spp. Oblong-elliptical, with a single longitudinal opening slit 80–95 × 7–8.5 µm 40–50 × 2–2.5 µm, with 1(–3)-septate, slightly tapering to both ends Amazonas, Brazil Johnston 2001
T. longissima On dead leaves of Bambusaceae sp. Oblong to sublinear, ends rounded, opening by a single, longitudinal slit 175–210 × 6–6.5 µm Approximately 120–130 µm long, but the detailed morphological characters were not seen Potaro-Siparuni region VII, Guyana Johnston 2001
T. mangiferae On dead leaves of Aucuba japonica and Mangifera indica Ellipsoidal, with a longitudinal opening split 80–90 × 5–6 µm 70–80 × 1 µm, filiform, lacking gelatinous sheath Java, Indonesia Koorders 1907; Li et al. 2014
T. meitanensis On dead culms of unidentified host Elliptical to oblong-elliptical, ends slightly acute to obtuse, opening by a single longitudinal split (98.5–)113–125.5(–131.5) × 6–7.5 µm 47–54.5 × 1.5–2.5 µm, filiform, gradually tapering towards both ends, aseptate, lacking gelatinous sheath Guizhou, China In this study
T. nematoidea On dead leaves of Gahnia sp. Elliptical to sublinear, with a single longitudinal opening slit 70–80 × 5–6.5 µm 30–35 × 1 µm, slightly tapering towards both ends, gently curved or sigmoid, 1-septate, lacking gelatinous sheath Northland, New Zealand Johnston 2001
T. nitens On leaves of Cyclobalanopsis myrsinifolia Suborbicular or broadly elliptical, straight or slightly curved, opening by a single longitudinal split 95–150 × 1–1.2 µm 68–115 × 0.8–1.2 µm, filiform, aseptate, round at the apex, slightly tapering towards the acute base, covered by a thin gelatinous sheath Anhui, China Chen et al. 2013
T. pandani On dead leaves of Pandanus sp. Oblong to oblong-elliptical, ends rounded, opening by a single longitudinal slit 100–120 × 5–6 µm 50–70 × 1–1.5 µm, lacking gelatinous sheath San Juan, Puerto Rico Johnston 2001
T. pandanicola On dead leaves of Pandanus sp. Elliptical, with rounded to subacute ends, opening by a longitudinal split 50–66 × 4–5 µm 55–78 × 1–2 µm, filiform, slightly tapering towards both ends, aseptate, lacking gelatinous sheath Prachuap Khiri Khan, Thailand Tibpromma et al. 2018
T. petrakii On fallen leaves of Smilax bracteata Elongate-elliptical, strongly curved or triangular, often coalesced, opening by a longitudinal split 85–110 × 4–5 µm (60–)70–85 × 0.8 µm, filiform, aseptate, covered by a thin gelatinous sheath Yunnan, China Song et al. 2012
T. rotundata On fallen leaves of Quercus sp. Elliptical, occasionally triangular, ends rounded, opening by a longitudinal split or occasionally by teeth 90–120 × 4–5.5 µm 70–90(–95) × 0.8–1 µm, filiform, aseptate, lacking gelatinous sheath Yunnan, China Song et al. 2012
T. sacchari On dead leaves and leaf bases of Saccharum officinarum Narrow-oblong to sublinear, with a single longitudinal opening split 90–100 × 5–7 µm 50–60 × 1.5 µm, lacking gelatinous sheath Hawaii, USA Johnston 2001
T. samuelsii On dead leaves of unidentified monocotyledon Oblong to sublinear, ends rounded, opening by a single longitudinal slit 125–140 × 7–8 µm (65–)75–90 × 2 µm, slightly tapering towards both ends, 1-septate, lacking gelatinous sheath Amazonas, Brazil Johnston 2001; 2003
T. sigmoideospora On dead fallen leaves of unidentified host Elliptical, ends rounded to subacute, opening by a single longitudinal split (93.5–)102–121 × 5–6 μm 79–95 × 5–2 μm, filiform, slightly tapering towards both ends, aseptate, lacking gelatinous sheath Guizhou, China In this study
T. simplex On fallen leaves of Trachelospermum jasminoides Elliptical to ovate, ends obtuse, rounded or slightly acute, opening by a single longitudinal split which is sometimes branched in the triangular ascomata 72–95(–105) × 4.8–5.2 µm (45–)56–82 × 1–1.2 µm, filiform, slightly tapering towards the rounded base, covered by a 0.8–1 µm wide gelatinous sheath Anhui, China Gao et al. 2012
T. stevensii On dead leaves of Vincentia sp. Oblong, ends rounded, opening by a single longitudinal slit 100–125 × 5–6 µm 60–80 × 1.5–2 µm, lacking gelatinous sheath Hawaii, USA Johnston 2001
T. thailandica On dead branch of unidentified host Elliptical, ends rounded to subacute, opening by a longitudinal split 80–105 × 3.4–6.6 µm 38–60 × 1–1.5 µm, filiform, slightly tapering towards both ends, aseptate, lacking gelatinous sheath Chiang Rai, Thailand Hyde et al. 2016
T. transversa On dead leaves of Pandanus sp. Elliptical or oblong-elliptical, ends slightly acute to obtuse, opening by a single longitudinal split 70–86 × 5–6 µm 45–68 × 1–1.2 µm, filiform, slightly tapering towards both ends, aseptate, covered by a 0.5 µm wide gelatinous sheath Hainan, China Li et al. 2015a

Terriera karsti J.F. Zhang & J.K. Liu, sp. nov.

Figure 3

Holotype

MFLU 18-2288.

Etymology

Refers to the karst landscape where the holotype was collected.

Description

Apothecia developing on dead branch, elliptical or oblong-elliptical in outline, ends slightly acute to obtuse. Apothecia surface black, matt or slightly glossy, moderately raising the substratum surface, opening by a single longitudinal split that extends to the ends of the apothecium (Fig. 3a, b). Lips absent. In median vertical section (Fig. 3d), apothecia deeply embedded in host tissue, with host cells becoming filled with fungal tissue as the apothecium develops. Covering stroma (Fig. 3c) 30–45 µm thick, composed of blackish-brown to black, thick-walled cells of textura angularis towards the exterior and several layers of pale to nearly hyaline, thin-walled cells towards the interior. Along the edge of the apothecial opening, there is a flattened, 12–20 µm thick extension adjacent to the covering stroma that is composed of strongly melanised tissue with no obvious cellular structure. Basal stroma 8–18 µm thick, dark brown or blackish-brown, composed of angular to globose, thick-walled cells, 2.5–4 µm diam. A triangular space between the covering stroma and basal stroma consists of thin-walled, nearly hyaline to grey-brown cells arranged in textura prismatica. Paraphyses 1–2 µm, filiform, hyaline, septate, gradually swollen or branching once at the apex, embedded in gelatinous sheaths. Asci (103–)110–122.5 × 5.5–7 µm (x¯ = 113 × 6 µm, n = 20), 8-spored, unitunicate, cylindrical, long stalk, thin-walled, apex truncate to somewhat round, J-, without circumapical thickening. Ascospores 55–66 × 1.5–2.0 µm (x¯ = 61 × 1.8 µm, n = 25), fascicle, but not coiled, filiform, gradually tapering toward the ends, hyaline, aseptate, smooth-walled, straight or slightly curved, lacking gelatinous sheath. Asexual morph: Not observed.

Figure 3. 

Terriera karsti a, b apothecia observed under the dissecting microscope c detail of covering stroma in vertical section d vertical section through an apothecium e, f asci in various states of maturity g apices of paraphyses h, i ascospores. Note: c–i mounted in water. Scale bar: 1 mm (a), 500 µm (b), 20 µm (c, e, f), 100 µm (d), 10 µm (g, i).

Culture characteristics

Colonies on PDA reaching 51 mm after 14 days at 25 °C, irregular in shape, cottony with moderately dense, fluffy aerial mycelium. At first, white, becoming slightly greyish in the centre, reverse side bronze in the centre and pale towards the edge.

Material examined

CHINA, Guizhou Province, Guiyang, Yunyan District, dead branch of unidentified ligneous plants, 6 May 2016, J.F. Zhang, SH-06 (MFLU 18-2288, holotype); ibid. (GZAAS 19-1720, isotype); ex-type living culture, GZCC 19-0047.

Notes

In the present study (Fig. 1), Terriera karsti is phylogenetically close to T. camelliicola and T. thailandica with moderate support (MPBP 63% and BYPP 1.00). Terriera karsti is not significantly distinguished from T. camelliicola, based only on morphological characters as they share similar-sized asci (110–122.5 × 5.5–7 µm vs. 85–120 × 5.5–6.5 µm) and ascospores (55–66 × 1.5–2 µm vs. 50–70 × 1 µm) (Johnston 2001). However, the ascospores of T. camelliicola are covered by a 0.5 µm wide gelatinous sheath, while this is not observed in T. karsti (Sharma 1982). In order to clarify their affinity, the recommendations of species delineation from Jeewon and Hyde (2016) were followed and the comparison of each gene region between these two taxa is processed and showed that there are 9/840 bp (1%) and 10/694 bp (14.4%) differences in LSU and mtSSU regions, respectively, while T. karsti can be easily differentiated from T. thailandica by its larger asci (110–122.5 × 5.5–7 µm vs. 80–105 × 3.4–6.6 µm) and ascospores (55–66 × 1.5–2 µm vs. 38–60 × 1–1.5 µm) (Hyde et al. 2016). A comparison of the LSU gene region between these two taxa has also been processed and the result showed that there are 3/838 bp (base pair) differences. Based on phylogenetic analyses, coupled with morphological distinction, Terriera karsti is introduced herein as a new species.

Terriera meitanensis J.F. Zhang & Z.Y. Liu, sp. nov.

Figure 4

Holotype

MFLU 18-2299.

Etymology

Referring to the locality of the holotype, Meitan County, Guizhou Province, China.

Description

Apothecia developing on dead stems (Fig. 4a), semi-immersed to superficial, elliptical or oblong-elliptical, ends slightly acute to obtuse, surface black, matt, raising the substratum surface, opening by a single longitudinal split that extends nearly the entire length (Fig. 4b, c). In median vertical section (Fig. 4d), apothecia deeply embedded in host tissue, with host cells becoming filled with fungal tissue as the apothecium develops. Covering stroma (Fig. 4e) 33–42 µm thick, composed of blackish-brown, thick-walled cells that are fused with host tissue in the outermost layers, becoming pale pigmented or nearly colourless towards the hymenium, thin-walled cells, arranged in textura angularis or textura globulosa. Along the upper edge of the apothecial opening, there is a flattened, 19–34 µm thick extension adjacent to the covering stroma that is composed of strongly melanised tissue with no obvious cellular structure. Basal stroma (Fig. 4g) 8–18 µm thick, dark-brown or blackish-brown, composed of angular to globose, thick-walled cells, 2.5–4 µm diam. Where the covering stroma meets the basal stroma, there is a triangular-shaped, 35–60 µm thick, tissue composed of thin-walled, hyaline to pale brown cells forming a textura prismatica (Fig. 4f). Subhymenium 12–16 µm thick, consisting of hyaline textura angularis to textura intricata. Paraphyses 1–2 µm, filiform, hyaline, septate, gradually swollen or branching once at the apex, embedded in gelatinous matrix, anastomosing at the base. Asci (98.5–)113–125.5(–131.5) × 6–7.5 µm (x¯ = 117 × 6.5 µm, n = 20), 8-spored, unitunicate, cylindrical, somewhat long-stalked, thin-walled, apex generally truncate, J-, without circumapical thickening. Ascospores 47–54.5 × 1.5–2.5 µm (x¯ = 50.5 × 2 µm, n = 35), fascicle, filiform, gradually tapering towards the ends, hyaline, aseptate, smooth-walled, straight or slightly curved, lacking a gelatinous sheath. Asexual morph: Not observed.

Figure 4. 

Terriera meitanensis a habit of apothecia on substrate b, c apothecia observed under the dissecting microscope in face view d vertical section through an apothecium e covering stroma f triangular space in section between the covering stroma and basal stroma g basal stroma h paraphyses with anastomoses amongst asci in various states of maturity i, j immature asci k, l ascospores. Note: d–l mounted in water. Scale bar: 1 cm (a), 1 mm (b), 500 µm (c), 100 µm (d), 10 µm (e, g, k, l), 30 µm (f), 20 µm (h–j).

Material examined

CHINA, Guizhou Province, Zunyi, Meitan County, dead stems of unidentified host, 28 August 2017, J.F. Zhang, MT-1 (MFLU 18-2299, holotype); ibid. (GZAAS 19-1731, isotype).

Notes

In our phylogenetic analysis (Fig. 1), Terriera meitanensis is placed in a robust clade with T. camelliicola, T. elliptica, T. karsti and T. thailandica by strong statistical support (MPBP 100% and BYPP 1.00). Terriera meitanensis has larger asci than T. camelliicola and T. thailandica, while the ascospores of T. meitanensis are smaller (Johnston 2001; Hyde et al. 2016). Both T. meitanensis and T. karsti share similar-sized asci, but T. karsti has larger ascospores (47–54.5 × 1.5–2.5 µm vs. 55–66 × 1.5–2.0 µm). Terriera meitanensis differs from T. elliptica by its obviously smaller asci (113–122.5 × 6–7.5 µm vs. 135–175 × 7–9 µm) and ascospores (47–54.5 × 1.5–2.5 µm vs. 60–85 × 1.5–2 µm) (Zhang et al. 2015). Moreover, the ascospores of T. camelliicola and T. elliptica are enveloped by a gelatinous sheath, respectively, while this is not observed in T. meitanensis. In addition, the comparison of the ITS gene region is processed between T. meitanensis and its closest species T. elliptica, based on the recommendations from Jeewon and Hyde (2016) and the results showed that there are 15/489 bp (3%) differences. Therefore, we introduce T. meitanensis herein as a new species, based on morphological and molecular evidence.

Terriera sigmoideospora J.F. Zhang & K.D. Hyde, sp. nov.

Figure 5

Holotype

MFLU 18-2297.

Etymology

Refers to its sigmoidal ascospores.

Description

Apothecia developing on fallen leaves, scattered, dark brown to black, matt, elliptical, sometimes 3-lobed or triangular, straight or slightly curved, ends rounded to subacute, strongly raising the surface of the substrate at maturity, opening by a single longitudinal split that extends almost the whole length of the apothecium (Fig. 5a, b). Immature apothecia appearing as a single dark brown protrusion, circular to slightly elongated. In median vertical section (Fig. 5d), apothecia 185–220 μm deep. Covering stroma (Fig. 5c) 20–25 μm thick near the centre of the apothecium, consisting of an outer layer of host cuticle, remains of epidermal and hypodermal cells filled with thick-walled, angular fungal cells and an inner layer of textura angularis to textura globulosa with 4–7 μm diam., dark brown, thick-walled cells, slightly thinner towards the edges, extending to the basal stroma, but conspicuously thicker towards the apothecial opening, with a 15–27 μm thick extension comprising highly melanised tissue with no obvious cellular structure. Excipulum moderately developed, closely adhering to the covering stroma and the extension, arising from the marginal paraphyses, becoming thinner towards the base. Basal stroma concave, 12–15 μm thick, composed of dark brown, thick-walled, angular cells. A triangular space between the covering stroma and basal stroma is composed of thin-walled, colourless cells that are vertically arranged in rows. Subhymenium 6–9 μm thick, flat, consisting of hyaline cells of textura intricata. Paraphyses filiform, hyaline, septate, gradually or suddenly swollen to 2.5 μm near the apex, covered by a thin gelatinous sheath, forming a 4–8 μm thick epithecium. Asci (93.5–)102–121 × 5–6 μm (x¯ = 108.5 × 5.5 µm, n = 20), 8-spored, unitunicate, cylindrical, apex tapering to round, thin-walled, J-, without circumapical thickening. Ascospores 79–95 × 1.5–2 μm (x¯ = 89.5 × 1.9 µm, n = 30), fascicle, filiform, sigmoid, tapering slightly towards the ends, hyaline, aseptate, guttulate, gelatinous sheath not observed. Asexual morph: Not observed.

Figure 5. 

Terriera sigmoideospora a, b apothecia observed under the dissecting microscope c section of covering stroma d median vertical section through an apothecium e immature ascus f paraphyses and asci in various states of maturity g, h ascospores. Note: c–h mounted in water. Scale bar: 1 mm (a), 500 µm (b), 100 µm (c), 20 µm (d–h).

Material examined

CHINA, Guizhou Province, Guiyang, dead leaves of unidentified host, 5 October 2016, J.F. Zhang, GZ-28 (MFLU 18-2297, holotype); ibid. (GZAAS 19-1729, isotype).

Notes

In the present phylogenetic analysis (Fig. 1), Terriera sigmoideospora is placed within Terriera and is related to T. houjiazhuangensis C.L. Hou & S.R. Hou by strong statistical support (MPBP 99% and BYPP 1.00). Terriera sigmoideospora shares similar-sized asci with T. houjiazhuangensis (102–121 × 5–6 μm vs. 103–128 × 4–6 μm), but has larger ascospores (79–95 × 1.5–2 μm vs. 73–82 × 0.6–0.9 μm) (Cai et al. 2020). Besides, the ascospores of T. houjiazhuangensis are enveloped by an inconspicuous gelatinous sheath, while this is not observed in T. sigmoideospora. In addition, the comparison of the ITS gene region between these two taxa has been processed and showed that there are 19/815 (2.3%) bp differences. Terriera pandanicola is sister to the above two taxa; however, it is significantly distinguished from T. sigmoideospora as its obviously smaller asci (50–66 × 4–5 μm vs. 102–121 × 5–6 μm) and ascospores (55–78 × 1–2 μm vs. 79–95 × 1.5–2 μm) (Tibpromma et al. 2018).

Discussion

The diversity of microfungi in many parts of the world is understudied. This is evident from the numerous new species being described from Asia and South America (Hyde et al. 2018, 2019a, 2020). With this in mind, we are studying the fungi of the Karst regions in China and Thailand, where we are also finding numerous new species (Zhang et al. 2016, 2017a, b, 2018, 2019). Our study is contributing to the knowledge of fungal diversity in the region, where species may also have biotechnological potential (Hyde et al. 2019b). Additionally, as Rhytismataceae is a relatively poorly studied group, we report on one new species from Hypoderma and three new Terriera species, thereby illustrating the diversity and potential for new discoveries of these fungi in Asia.

Hypoderma, a large genus in Rhytismataceae, is a complicated group. There are only a few species in this genus with sequence data, but these have shown the group to be polyphyletic (Lantieri et al. 2011; Wang et al. 2013). This is also true of the phylogenies in this study (Fig. 1). Hypoderma is morphologically similar to Lophodermium and they mainly differ on the basis of ascospore shape as the former have elliptical to cylindrical-fusiform ascospores, while the latter has filiform ascospores (Powell 1974). However, there are no molecular studies that provide a natural classification for these two genera, even though more than 35 species have been synonymized under Lophodermium (Index Fungorum 2020). Fresh collections and molecular sequences are required to move toward a revision of these genera.

Terriera is one of the few genera in Rhytismataceae that can be considered a monophyletic group, based on distinctive morphology and phylogenetic characterisation (Zhang et al. 2015). Our molecular analyses corroborate this. However, there are only nine taxa with available sequences in GenBank and most of Terriera species were established, based only on morphological features (Yang et al. 2011; Gao et al. 2012; Song et al. 2012; Zhou et al. 2012; Chen et al. 2013; Li et al. 2015b; Lu et al. 2015; Zhang et al. 2015; Cai et al. 2020). In the latest study (Cai et al. 2020), T. pandanicola was distant from Terriera in ITS analysis, but included in this group on the basis of concatenated LSU-mtSSU sequence data. Cai et al. (2020) indicated that this taxon should be revised in a future study. Based on their suggestion, we checked the sequence data of T. pandanicola and found that the ITS sequence of this species is misidentified as it is not a related Terriera or even a Rhytismataceae species in BLASTn results. However, the newly generated available sequences (ITS and mtSSU) of T. pandanicola have been uploaded in GenBank and included in our phylogenetic analysis and the results indicated that it is a unique species in Terriera in the present study (Fig. 1).

Acknowledgements

Kevin D. Hyde thanks the Thailand Research grants entitled “The future of specialist fungi in a changing climate: baseline data for generalist and specialist fungi associated with ants, Rhododendron species and Dracaena species” (Grant No. DBG6080013) and “Impact of climate change on fungal diversity and biogeography in the Greater Mekong Subregion” (Grant No. RDG6130001). Jason M. Karakehian is thanked for revising the manuscript. Dr. Shaun Pennycook (Manaaki Whenua Landcare Research, New Zealand) is gratefully thanked for advising on the fungal names. Dr. Saowaluck Tibpromma is thanked for updating the new sequences of T. pandanicola. Jin-Feng Zhang would like to thank Dr. Peter R. Johnston for providing literature and suggestions.

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Supplementary material

Supplementary material 1 

Dataset for molecular analyses

Jin-Feng Zhang, Jian-Kui Liu, Kevin D. Hyde, Anusha H. Ekanayaka, Zuo-Yi Liu

Data type: phylogenetic

Explanation note: The dataset of combined of LSU_ITS_mtSSU to build the phylogenetic tree.

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.
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