132 urn:lsid:arphahub.com:pub:6d16bb82-189e-56df-a05d-907690ec8db9 IMA Fungus imafungus 2210-6340 2210-6359 Pensoft Publishers 10.3897/imafungus.17.187200 187200 Research Article Ascomycota Taxonomy Asia Phylogenetic analysis of the genus Taphrina: proposal of one new genus and seven new species (Taphrinales, Taphrinomycetes) from China Chai Chun-Yue https://orcid.org/0000-0001-7753-6223 1 2 Cai Dan-Yang https://orcid.org/0009-0002-6866-7322 1 Song Bing-Yan https://orcid.org/0009-0000-6802-5077 1 Wang Rui-Xiu https://orcid.org/0009-0009-5141-7096 1 Hui Feng-Li fenglihui@yeah.net https://orcid.org/0000-0001-7928-3055 1 2 School of Life Science, Nanyang Normal University, Nanyang 473061, China Research Center of Henan Provincial Agricultural Biomass Resource Engineering and Technology, Nanyang Normal University Nanyang China https://ror.org/01f7yer47 Research Center of Henan Provincial Agricultural Biomass Resource Engineering and Technology, Nanyang Normal University, Nanyang 473061, China School of Life Science, Nanyang Normal University Nanyang China https://ror.org/01f7yer47

Corresponding author: Feng-Li Hui (fenglihui@yeah.net)

Academic editor: C. Alisha Quandt

 2026 24 08 2026 17 e187200 3C8A0830-A668-5A85-AF36-F1FCA155BB0A 01 02 2026 10 08 2026 Chun-Yue Chai, Dan-Yang Cai, Bing-Yan Song, Rui-Xiu Wang, Feng-Li Hui 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.

Taphrina, a genus within the family Taphrinaceae and the order Taphrinales, is a globally distributed group of plant-associated fungi. However, only three species of this genus have been previously reported in China. To enhance the understanding of species diversity within the Taphrina genus in China, sixteen yeast strains, isolated from leaf surfaces across various locations in China and initially identified as members of Taphrina, were examined from both phylogenetic and phenotypic perspectives. From these isolates, one new genus, Srisukozyma, is described to accommodate two new species, Srisukozyma ovata and Sr. musae, along with five additional new species Taphrina aspidistrae, T. cunninghamiae, T. foliicola, T. fujianensis, and T. guizhouensis which we describe. The descriptions of these taxa are supplemented by illustrations and results from phylogenetic analyses. Additionally, phenotypic comparisons with related genus and species are provided. This study contributes to the broader understanding of ascomycetous yeast diversity in natural environments and sets the stage for future taxonomic and ecological investigations.

 Ascomycota phylogenetic analysis Taphrinomycotina taxonomy Citation

Chai C-Y, Cai D-Y, Song B-Y, Wang R-X, Hui F-L (2026) Phylogenetic analysis of the genus Taphrina: proposal of one new genus and seven new species (Taphrinales, Taphrinomycetes) from China. IMA Fungus 17: e187200. https://doi.org/10.3897/imafungus.17.187200

 Introduction

Taphrina, the type genus of the family Taphrinaceae, was established by Fries in 1832 with T. populina as the type species (Fonseca and Rodrigues 2011). Initially, the genus was described as a sexual ascomycete fungus within the “Archiascomycetes” (Nishida and Sugiyama 1994), later being placed in the class Taphrinomycetes, which was formally proposed for archiascomycete lineages by Eriksson and Winka (1997). The asexual state of Taphrina was originally classified under the genus Lalaria (Eriksson and Winka 1997; Inácio et al. 2004). However, molecular analyses have unequivocally placed previously recognized Lalaria anamorphs within the monophyletic genus Taphrina, which belongs to the order Taphrinales (Sjamsuridzal et al. 1997; Rodrigues and Fonseca 2003; Fonseca and Rodrigues 2011). Therefore, Selbmann et al. (2014) emended the genus Taphrina and, in accordance with the International Code of Nomenclature for Algae, Fungi, and Plants (McNeill et al. 2012), proposed that each asexual morph (previously placed in Lalaria) be combined with its corresponding sexual morph into a single species within the genus Taphrina.

Most species of the genus Taphrina exhibit a dimorphic lifestyle. The sexual morph is filamentous, strictly biotrophic, and forms asci within the infected plant tissues (Rodrigues and Fonseca 2003; Fonseca and Rodrigues 2011; Petrýdesová et al. 2013). In contrast, the asexual morph is yeast-like, saprobic, reproduces by budding, and is easily cultured on artificial media (Tsai et al. 2014; Petrýdesová et al. 2016). Cultures are often pigmented, appearing cream, yellowish, or pinkish-cream in color, with pigmentation intensifying as they age (Selbmann et al. 2014). Notably, cultures of many Taphrina species are composed of yeast forms that have been isolated from infected tissues of the respective host plants using the spore-fall method (Fonseca and Rodrigues 2011).

Members of the genus Taphrina are parasitic pathogens of various vascular plants, including economically important fruit trees, shrubs, herbs, and some ferns (Mix 1949; Cissé et al. 2013; Tsai et al. 2014). Specific examples include T. populina on poplars (Populus spp.) (Robinson et al. 2011; Tsai et al. 2014), T. ulmi on elms (Ulmus spp.) (Bond 1956), T. wiesneri on flowering cherry (Prunus yedoensis) (Tsai et al. 2014; Lee et al. 2025), and T. caerulescens on oaks (Quercus spp.) (Sutton and Hodges 1990). T. deformans is the most extensively studied species in the genus due to its significance as a plant pathogen worldwide, with all known peach and nectarine cultivars being susceptible to infection (Cissé et al. 2013; Oh et al. 2020). One of the most notable symptomatic effects is the severe distortion and reddening of the leaves (Kern and Naef-Roth 1975). These symptoms are typically linked to the production of indole-3-acetic acid (IAA) by T. deformans (Yamada et al. 1990; Tsai et al. 2014). Another significant species is T. flavorubra, the causative agent of deformed fruits, or plum pocket, in plums (Rodrigues and Fonseca 2003; Cissé et al. 2013). Infected fruits exhibit abnormal enlargement, deformation, and a spongy texture. Additionally, T. betulina significantly reduces the mean height and diameter of infected Betula pubescens (birch trees) (Spanos and Woodward 1994), potentially leading to losses in timber production, especially in short-rotation forestry (Selbmann et al. 2014).

Taphrina is an ancient, species-rich ascomycete genus, with nearly 100 scientific names recorded under it (Wang et al. 2020). Unfortunately, approximately 70% of the species described have not been deposited in culture collections. Rodrigues and Fonseca (2003) clarified the generic boundaries of Taphrina using a morpho-molecular approach. Due to low DNA sequence divergence and the absence of available strains, the number of accepted Taphrina species was reduced to 28 (Fonseca and Rodrigues 2011). Since then, nine additional species have been recognized based on phenotypic and phylogenetic analyses (Petrýdesová et al. 2013, 2016; Selbmann et al. 2014). Currently, 38 valid species are accepted in the genus Taphrina (Petrýdesová et al. 2016). However, only four Taphrina species—T. cerasi, T. deformans, T. mume, and T. pruni—have been reported in China (Wang et al. 2022). In the present study, strains of Taphrina-like taxa were isolated from different locations in China. To clarify their taxonomic status, both phylogenetic and phenotypic analyses were conducted. Five new species are described within Taphrina, and one new genus, accommodating two new species, is introduced within the Taphrinomycetes.

 Materials and methods Sample collection and yeast isolation

Fresh leaves were collected from Fujian, Guizhou, Hainan, and Henan Provinces of China. Yeast strains were isolated from the leaf surfaces using an improved spore-fall method, as described by Nakase and Takashima (1993). In brief, the leaves were cut into small pieces and attached to the inner lid of a Petri dish using a thin layer of petroleum jelly. The dish contained yeast extract-malt extract (YM) agar (0.3% yeast extract, 0.3% malt extract, 0.5% peptone, 1% glucose, and 2% agar), with 0.01% chloramphenicol added to prevent bacterial growth. The plates were incubated at 20 °C and monitored daily for colony formation. Colonies were selected and purified by streaking onto separate YM agar plates. After purification, yeast strains were suspended in YM broth containing 20% (v/v) glycerol and stored at −80 °C.

 Phenotypic examination

Phenotypic characteristics were determined following the standard methods described in Kurtzman et al. (2011). Colony and cell morphology: Colony morphology was examined after 7 days of incubation at 20 °C on YM agar. Cell morphology was observed using a LEICA DM2500 microscope (LEICA, Wetzlar, Germany) after 3 days of growth in YM broth at 20 °C, and analyzed with LASV4.13 software. Glucose fermentation: Fermentation was tested using Durham fermentation tubes incubated at 20 °C for 14 days; gas production indicated a positive reaction. Carbon assimilation: Tests were performed in liquid medium containing 0.67% yeast nitrogen base (YNB) and 0.5% of the test carbon compound. Incubation was at 20 °C for 28 days; growth (turbidity) was recorded as positive. Nitrogen assimilation: Starved inoculum was prepared by growing yeasts in YNB broth for 48 h, followed by centrifugation, two washes with sterile distilled water, and resuspension in sterile water. The test medium consisted of 1.77% yeast carbon base (YCB) and the specified concentration of the test nitrogen compound. After 28 days of incubation at 20 °C, growth was evaluated by turbidity. Pseudohyphae formation: The Dalmau plate method on corn meal agar (CMA: 2.5% corn meal infusion and 2% agar) was used, with sterile coverslips creating an anaerobic environment. Plates were incubated at 20 °C for 14 days and examined microscopically. Sexual reproduction: Single strains or strain pairs were inoculated on CMA, 5% malt extract (ME) agar (5% malt extract and 1.5% agar), Fowell’s acetate agar (0.5% sodium acetate and 2% agar), Gorodkowa agar (0.1% glucose, 0.5% sodium chloride, 1% peptone, and 2% agar), and V8 agar (10% V8 juice and 2% agar), and incubated at 20 °C for up to two months; asci and ascospores were examined periodically. Additional growth tests: Growth at different temperatures (15, 20, 25, 30, 35, and 37 °C) was assessed on YM agar. Growth in vitamin-free medium, tolerance to 1% acetic acid, 0.01% cycloheximide, and 10% NaCl plus 5% glucose, starch-like polysaccharide production (Lugol’s iodine), urease activity (Christensen’s urea agar), and Diazonium Blue B (DBB) reaction were all performed according to the standard protocols in Kurtzman et al. (2011). All novel taxonomic descriptions and proposed names were deposited in the MycoBank database (http://www.mycobank.org; accessed 10 December 2025).

 Molecular sequencing

Genomic DNA was extracted from yeast strains using the Ezup Column Yeast Genomic DNA Purification Kit following the manufacturer’s instructions (Sangon Biotech, Co., Ltd., Shanghai, China; Cat. No. B518225-0100). The primer pairs ITS5 (5’-GGAAGTAAAAGTCGTAACAAGG-3’)/ITS4 (5’-TCCTCCGCTTATTGATATGC-3’) (White et al. 1990), NL1 (5’-GCATATCAATAAGCGGAGGAAAAG-3’)/NL4 (Kurtzman and Robnett 1998), and SSU1 (5’-CTGGTTGATCCTGCCAGTAG-3’) /SSU5 (5’-TGATCCTTCTGCAGGTTCACCTAC-3’) (Petrýdesová et al. 2013) were selected to amplify the internal transcribed spacer (ITS) region, the D1/D2 domain of the large subunit (LSU) rRNA gene, and the partial mitochondrial small rRNA subunit (mtSSU) region, respectively. PCR amplification was carried out in a 25 µL reaction volume containing 9.5 µL of ddH2O, 12.5 µL of 2 × Taq PCR Master Mix with blue dye (Sangon Biotech Co., Shanghai, China), 1 µL of DNA template, and 1 µL of each primer. The PCR protocols were as follows: for the ITS region, an initial denaturation at 95 °C for 3 min, followed by 34 cycles at 94 °C for 40 s, 57.2 °C for 45 s, and 72 °C for 1 min, with a final extension at 72 °C for 10 min; for the LSU region, an initial denaturation at 94 °C for 1 min, followed by 34 cycles at 94 °C for 30 s, 55.2 °C for 1 min, and 72 °C for 1.5 min, with a final extension at 72 °C for 10 min (Chai et al. 2023); for the mtSSU region, an initial denaturation at 94 °C for 3 min, followed by 36 cycles at 94 °C for 45 s, 50 °C for 2 min, and 72 °C for 2 min, with a final extension at 72 °C for 5 min (Petrýdesová et al. 2013). The PCR products were sequenced using the same primers at Sangon Biotech Co., Ltd., Shanghai, China. Forward and reverse sequences were manually edited using BioEdit v7.1.3.0 (Hall 1999) by removing primer sequences, trimming low-quality ends based on chromatogram quality scores (Phred < 20), and resolving ambiguous base calls manually. The edited forward and reverse reads were then assembled into consensus sequences using the same software. Sequence similarity searches were performed using the BLASTN 2.2.19 algorithm (Zhang et al. 2000) against the GenBank database. All newly generated sequences were deposited in GenBank (https://www.ncbi.nlm.nih.gov/genbank/).

 Phylogenetic analyses

Phylogenetic analyses were based on combined sequences of the ITS, LSU, and mtSSU regions. Sequences from various Taphrinomycotina species retrieved from GenBank were combined with those generated in our study. Information on the studied yeasts and their corresponding GenBank accession numbers is provided in Table 1. Sequences for each locus were aligned using MAFFT v.7.110 (Katoh and Standley 2013) with the G-INI-I option (Katoh et al. 2005). The resulting alignments were visualized using MEGA v.11 (Tamura et al. 2021). To avoid inclusion of poorly aligned or highly gapped positions, the alignments were manually trimmed by removing ambiguous regions at both ends as well as any positions with >90% gaps or unresolved bases; all trimming was performed under visual inspection within MEGA v.11. Phylosuite v.1.2.2 (Zhang et al. 2020) was used to concatenate the trimmed alignments from the different loci. The best-fit evolutionary models for the alignments were estimated using jModelTest (Guindon and Gascuel 2003; Posada 2008) under the Akaike information criterion.

The accession numbers of studied yeasts and reference sequences retrieved from the GenBank database.

Taxa name Strain no. Substrate or host Location GenBank accession no. References
ITS LSU D1/D2 mtSSU
Novakomyces olei NCAIM Y.02187T Olive oil Spain NR_172188 MG250349 – Čadež et al. 2021
Protomyces arabidopsidicola DSM 110145T Arabidopsis thaliana (Leaf) Finland LT602858 MK934482 – Wang et al. 2021
Protomyces inouyei NRRL YB-4354T Crepis sp. (Pedicel) Japan NR_121206 NG_042406 KU134844 Petrýdesová et al. 2016
Protomyces inundatus NRRL Y-6349T – England MK937057 U76528 – Kurtzman and Robnett 1998
Protomyces gravidus NRRL Y-17093T Bidens cernua USA MK937055 NG_064601 – Kurtzman and Robnett 1998
Protomyces lactucaedebilis NRRL YB-4353 T Lactuca debilis (Leaf) Japan MK937058 U84343 – Kurtzman and Robnett 1998
Protomyces macrosporus NRRL Y-12879T – USA MK937059 U94939 – Kurtzman and Robnett 1998
Protomyces pachydermus NRRL YB-4355T Taraxacum officinale England MK937060 NG_064602 – Kurtzman and Robnett 1998; Petrýdesová et al. 2016
Saitoella coloradoensis NRRL YB-2330T Insect frass USA NR_137778 NG_059942 JN161163 Kurtzman and Robnett 2012; Petrýdesová et al. 2016
Saitoella complicata NRRL Y-17804T Soil Bhutan NR_111655 NG_027621 KU134846 Goto et al. 1987; Petrýdesová et al. 2016
Savitreella phatthalungensis PYCC 9005T Ananas comosus (Leaf) Thailand MW879743 MW876306 – Nutaratat et al. 2022
Srisukozyma musae NYNU 24437 T Musa nana China PP849688 PP837701 PX939036 This study
Srisukozyma musae NYNU 244126 Musa nana China PX070522 PX070523 PX939145 This study
Srisukozyma musae NYNU 244111 Musa nana China PX070525 PX070524 PX939143 This study
Srisukozyma musae NYNU 24449 Musa nana China PX070527 PX070526 PX939037 This study
Srisukozyma ovata NYNU 2411186T Polygonum chinense China PV843161 PV839685 PX939146 This study
Srisukozyma ovata NYNU 2411221 Polygonum chinense China PX070537 PX070536 PX939149 This study
Taphrina alni CBS 683.93T Alnus incana Austria AF492077 AF492024 KU134812 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina americana CBS 331.55T Betula fontinalis USA NR_155874 AF492025 KU134813 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina antarctica CCFEE 5198T Sandstone Antarctica NR_132870 NG_059477 – Selbmann et al. 2014
Taphrina arrabidae PYCC 5735T Pistacia lentiscus Portugal PX671092 NG_059909 – Inácio et al. 2004
Taphrina aspidistrae NYNU 2411134T Aspidistra elatior China PV839642 PV839641 PX939148 This study
Taphrina aspidistrae JS-42 – Korea JF706660 JF706659 – Direct from Genbank
Taphrina betulina CBS 119536T Betula ‘intermedia’ Sweden AF492080 AF492027 KU134847 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina bullata CBS 1278 T Pyrus communis Slovakia KC491200 JN997390 KC494909 Petrýdesová et al. 2013; Petrýdesová et al. 2016
Taphrina caerulescens CBS 351.35T Quercus marylandica USA NR_155875 NG_057691 KU134815 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina carpini CBS 102169T Quercus pyrenaica Slovakia NR_119488 NG_042399 KU134814 Inácio et al. 2004; Petrýdesová et al. 2016
Taphrina communis CBS 352.35T Prunus americana (Leaf and Fruit) USA NR_160070 NG_070567 KU134816 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina confusa CBS 375.39T Prunus virginiana (Flower and Fruit) USA NR_155876 AF492035 – Rodrigues and Fonseca 2003
Taphrina cunninghamiae NYNU 232134T Cunninghamia lanceolata China OQ851904 OQ851903 PX939040 This study
Taphrina cunninghamiae NYNU 232156 Cunninghamia lanceolata China PX070522 PX070523 PX939039 This study
Taphrina dearnessii CBS 334.55T Acer rubrum (Leaf) USA NR_172284 AF492037 – Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina deformans CBS 356.35T Prunus persica Netherlands NR_160071 MH867217 KU134817 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina epiphylla CBS 111109T Alnus incana Slovakia AF492096 AF492039 KU134818 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina flavorubra CBS 377.39T Prunus susquehanae (Fruit) USA NR_160073 MH867541 KU134819 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina foliicola NYNU 2211137T Carpinus turczaninowii China OP959653 OP959502 PX939147 This study
Taphrina foliicola NYNU 221115 Carpinus turczaninowii China OP954718 OP954686 PX939038 This study
Taphrina foliicola NYNU 236334 Cornus officinalis China PX063790 PX070528 PX939043 This study
Taphrina foliicola NYNU 236258 Carpinus turczaninowii China PX063794 PX070529 PX939042 This study
Taphrina foliicola CGMCC 2.2537 – China PQ309310 PQ309490 – Direct from Genbank
Taphrina fujianensis NYNU 224115T Dicranopteris sp. China OP278686 OP278685 PX939041 This study
Taphrina fujianensis CGMCC 2.5682 – China PQ309311 PQ309491 – Direct from Genbank
Taphrina guizhouensis NYNU 24380T Cunninghamia lanceolata China PV839639 PV839638 PX939035 This study
Taphrina guizhouensis NYNU 243137 Rhododendron simsii China PX070534 PX070535 PX939144 This study
Taphrina gei-montani CBS 14159T Geum montanum Slovakia KU134800 – KU134828 Petrýdesová et al. 2016
Taphrina inositophila PYCC 5733T Acer monspessulanum Portugal NR_111147 AY239199 – Inácio et al. 2004
Taphrina insititiae CBS 12782T Prunus insititia Slovakia KC491202 JN997391 KC494911 Petrýdesová et al. 2013; Petrýdesová et al. 2016
Taphrina johansonii CBS 378.39T Populus tremuloides USA KC511055 NG_057692 KC494916 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina kurtzmanii PYCC 5737T Cistus albidus Portugal PX671093 NG_059910 – Inácio et al. 2004
Taphrina letifera CBS 335.55T Acer spicatum (Leaf) USA NR_155877 NG_057693 KU134820 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina mirabilis CBS 357.35T Prunus angustifolia USA NR_155878 NG_070568 – Rodrigues and Fonseca 2003
Taphrina padi CBS 693.93T Prunus padus (Fruit) Germany NR_155879 AF492048 KC494912 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina populina CBS 337.55T Populus nigra (Leaf) Sweden NR_145337 NG_057694 KU134821 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina populi-salicis CBS 419.54T Populus trichocarpa (Leaf) USA NR_155881 NG_057695 KU134822 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina pruni CBS 119537T Prunus domestica Slovakia AF492111 AF492056 KU134823 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina pruni-subcordatae CBS 381.39T Prunus subcordata Slovakia NR_155880 NG_070569 – Rodrigues and Fonseca 2003
Taphrina rhizophora CBS 1278T Populus alba Slovakia KC491201 KC491203 KC494917 Petrýdesová et al. 2013; Petrýdesová et al. 2016
Taphrina robinsoniana CBS 382.39T Alnus incana Slovakia NR_172369 NG_057696 KU134824 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina sacchari CBS 119738T Acer saccharum USA AF492117 AF492061 – Rodrigues and Fonseca 2003
Taphrina sadebeckii CBS 102170T Alnus glutinosa Germany NR_155882 NG_057697 KU134825 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina tormentillae CBS 339.55T Potentilla canadensis USA NR_155883 NG_057698 KU134845 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina tosquinetii CBS 276.28T Alnus glutinosa (Leaf) Europe NR_155884 NG_057699 KU134826 Rodrigues and Fonseca 2003; Petrýdesová et al. 2016
Taphrina ulmi CBS 420.54T Ulmus rubra (Leaf) USA NR_155885 NG_057700 KU134827 Rodrigues and Fonseca 2003
Taphrina veronaerambellii PYCC 5734T Acer monspessulanum Portugal NR_111148 NG_059911 NG_059911 Inácio et al. 2004; Petrýdesová et al. 2016
Taphrina vestergrenii CBS 679.93T Dryopteris clix-mas Europe NR_155886 NG_057701 – Rodrigues and Fonseca 2003
Taphrina virginica CBS 340.55T Ostrya virginiana USA AF492125 AF492071 – Inácio et al. 2004
Taphrina viridis CCY 58-9-1T Alnus alnobetula Slovakia PQ013128 PQ013143 PQ013155 Petrýdesová et al. 2016
Taphrina wiesneri CBS 275.28T Prunus avium – NR_160065 AF492072 KC494915 Rodrigues and Fonseca 2003
Schizosaccharomyces cryophilus CBS 11777T Osmia rufa (Feed) Denmark NR_121468 GU470882 NC_040930 Kurtzman and Robnett 1998
Schizosaccharomyces japonicus CBS 354T Strawberry wine Japan NR_121199 U94943 – Kurtzman and Robnett 1998
Schizosaccharomyces osmophilus CBS 15793T Osmia florisomnis (Bee bread) Germany MK589403 MK253005 NC_071386 Kurtzman and Robnett 1998
Schizosaccharomyces pombe CBS 356T Beer Africa MK749863 NG_042649 AF442355 Kurtzman and Robnett 1998, 2003
Saccharomyces cerevisiae CBS 1171T – France AB018043 JQ689017 KX657745 Kurtzman and Robnett 1998; Sulo et al. 2017

Note. T, type strain. Species obtained in this study are in bold.

Maximum Likelihood (ML) and Bayesian Inference (BI) analyses were performed using RAxML v.8.2.3 (Stamatakis 2014) and MrBayes v.3.2.7a (Ronquist et al. 2012), respectively. For the ML analysis, a GTRGAMMA model of evolution was applied, and bootstrap (BS) values were assessed through 1,000 rapid bootstrap replicates. In the BI analysis, six Markov Chain Monte Carlo (MCMC) chains were run simultaneously for 50 million generations under the best-fit evolutionary models selected using jModelTest (Guindon and Gascuel 2003; Posada 2008), with trees sampled every 1,000 generations. The first 25% of the generated trees were discarded as burn-in, and the remaining trees were used to estimate Bayesian posterior probabilities (BPPs) for the clades. Each tree was visualized with BS values and BPPs in Figtree v.1.4.3 (Andrew 2016).

 Results Molecular phylogeny

A total of 73 sequences, including the outgroup Saccharomyces cerevisiae CBS 1171, were used in the phylogenetic analyses to determine the taxonomic placement of the isolates obtained in this study. The concatenated dataset of the three regions across the 73 samples had a total length of 1,979 bp, consisting of 598 bp for the ITS region, 574 bp for the LSU region, and 807 bp for the mtSSU region. The phylogeny generated from the combined ITS, LSU, and mtSSU dataset strongly supported Taphrinales as an independent order (BS = 100%, BPPs = 1.0). Within Taphrinales, five new lineages emerged from Taphrina, and two new independent clades clustered with Savitreella phatthalungensis, with full support.

Ten strains, NYNU 2211137, NYNU 221115, NYNU 236334, NYNU 236258, NYNU 224115, NYNU 24380, NYNU 243137, NYNU 2411134, NYNU 232134, and NYNU 232156, formed five groups within the genus Taphrina, represented by NYNU 2211137, NYNU 224115, NYNU 24380, NYNU 2411134, and NYNU 232134 (Fig. 1). Group NYNU 2211137, containing four strains with one nucleotide substitution in the ITS region, had identical LSU sequences with strain CGMCC 2.2537 and two nucleotide ITS sequences difference, which indicated that they were conspecific. This group formed a clade clustered with T. carpini, T. inositophila, and T. virginica with significant support (Fig. 1) and differed from those three known species in this clade by more than three nucleotide substitutions (0.5%) in the LSU region and over 35 nucleotide mismatches (5.9%) in the ITS region. Based on these results, we conclude that the group NYNU 2211137 corresponds to a novel species, for which we propose the name Taphrina foliicola sp. nov.

10.3897/imafungus.17.187200.figure1 54A2C631-26D9-5EB5-B922-36CCBA0E8D53

Phylogenetic positions of the newly studied strains within Taphrinomycotina, inferred from the combined ITS, LSU, and mtSSU dataset. The tree topology was generated using the maximum likelihood method, with bootstrap values and Bayesian posterior probabilities (above 50% and 0.95, respectively) indicated at the nodes. The tree was rooted with Saccharomyces cerevisiae CBS 1171. Sequences from type strains are marked with a (T), and the newly proposed species are highlighted in bold.

https://binary.pensoft.net/fig/1753477

Strain NYNU 224115 clustered with strain CGMCC 2.5682, sharing identical LSU sequences and similar ITS sequences (with two nucleotide differences), which indicated that they belong to same species. Group NYNU 24380, consisting of two strains with identical sequences, indicating that they are also conspecific. These two groups formed two separate branches within the genus Taphrina (Fig. 1). They differed from each other by nine nucleotide substitutions (1.6%) and 40 nucleotide mismatches (6.3–6.6%) in the LSU and ITS regions, respectively. They also differed from other Taphrina species by more than nine nucleotide substitutions (1.6%) in the LSU region and more than 57 nucleotide substitutions (9.1%) in the ITS region. Based on these differences, we conclude that these two groups represent two novel Taphrina species. Therefore, we propose Taphrina fujianensis sp. nov. and Taphrina guizhouensis sp. nov. to accommodate the groups NYNU 224115 and NYNU 24380, respectively.

Strain NYNU 2411134 clustered with strain JS-42, sharing identical ITS sequences and similar LSU sequences (with a three nucleotides difference), suggesting that they are the same species. Groups NYNU 232134, including two strains with identical sequences, and 2411134 were closely related to T. arrabidae (Fig. 1). These two groups differed from T. arrabidae by 14 nucleotide substitutions (2.4%) in the LSU region and by greater than 38 nucleotide mismatches (6.1%) in the ITS region. Groups NYNU 232134 and 2411134 showed 15 nucleotide substitutions (2.6%) in the LSU region and more than 9% nucleotide mismatches in the ITS region. Based on these findings, we propose two new species, Taphrina aspidistrae sp. nov. and Taphrina cunninghamiae sp. nov., to accommodate the groups NYNU 2411134 and NYNU 232134.

Six strains, NYNU 24437, NYNU 244126, NYNU 244111, NYNU 24449, NYNU 2411186, and NYNU 2411221, formed two distinct branches closely related to Savitreella (Fig. 1), a new genus recently described by Nutaratat et al. (2022). The group represented by NYNU 24437 differed from the group represented by NYNU 2411186 by 33 nucleotide substitutions (5.7%) in the LSU region and by 85–88 nucleotide mismatches (13.4–13.9%) in the ITS region. A BLASTn search of the LSU sequences revealed that the two groups showed 88–90% similarity to genera within the Taphrinales, such as Taphrina, Protomyces, and Savitreella. However, only 25–35% of the ITS sequence of these groups matched taxa from these genera when using ITS sequences as the query. The taxonomic thresholds predicted by Vu et al. (2016) to discriminate current yeast genera were 97.11% for LSU and 96.31% for ITS. Given the low sequence similarity to any known taxa and strong phylogenetic support within Taphrinales, it is appropriate to establish one new genus Srisukozyma within Taphrinales. Therefore, we propose Srisukozyma musae sp. nov. and Srisukozyma ovata sp. nov. for the groups NYNU 24437 and NYNU 2411186, respectively.

 Taxonomy Fungi Haplotaxida Taphrinaceae EAF3CCC7-3F36-5478-A863-274704D9DEF7 Taphrina aspidistrae 861665 C.Y. Chai & F.L. Hui sp. nov. Fig. 2A, B Etymology.

The specific epithet aspidistrae refers to Aspidistra, the plant genus from which the type strain was isolated.

10.3897/imafungus.17.187200.figure2 BADB93F6-3AD6-561B-908F-3FA4D4F17706

The streak culture grown on YM agar for 7 d at 20 °C and vegetative cells grown in YM broth for 3 d at 20 °C. A, B. T. aspidistrae NYNU 2411134. C, D. T. cunninghamiae NYNU 232134. E, F. T. fujianensis NYNU 224115. G, H. T. guizhouensis NYNU 24380. I, J. T. foliicola NYNU 2211137. Scale bars: 10 μm.

https://binary.pensoft.net/fig/1753478 Diagnosis.

In the multi-gene phylogenetic analysis, T. aspidistrae clustered as sister to T. cunninghamiae and T. arrabidae with high support values (Fig. 1). T. aspidistrae differed from the closely related species T. cunninghamiae and T. arrabidae by 14–15 nucleotide substitutions (2.4–2.6%) in the LSU region and by 40–57 nucleotide mismatches (6.4–9%) in the ITS region (Suppl. material 2: table SS1). Physiologically, T. aspidistrae can be distinguished from T. cunninghamiae in its inability to assimilate trehalose, maltose, ethanol, and DL-lactate. It can also be distinguished from T. arrabidae in its ability to assimilate inulin, methyl-α-D-glucoside, L-arabinose, D-ribose, ribitol, and D-glucitol as well as grow in vitamin-free medium (Table 2; Suppl. material 2: table SS1).

Physiological and biochemical characteristics that differ between the new species and closely related species.

Characteristics 1 2* 3* 4* 5 6 7* 8* 9 10* 11 12 13 14*
Carbon assimilation
Inulin d w + – + + – – + – + – + N
Sucrose + + + + + + – – + + + + + +
Raffinose d/w – + + – – – – – – – + d –
Trehalose + + + + + w s + – – + + + +/w
Maltose + + + + + d – – – – + d/w + +
Melezitose + + + + + w – – w + + + + +
Methyl-α-D-glucoside – – + d/– + d – – d/w – + – d/w –
Cellobiose + + + + + d s + w + + – – S
Salicin d/w + + d + w – + w S + d/w d/w W
L-Sorbose – – – –/d – – – v – – – d – –
L-Rhamnose – – – – – – – – – – + – – –
D-Xylose + + + v + w s + + + + + d +/s
L-Arabinose – + + v + d/w – – d/w – + – d/w –
D-Arabinose – w + + + + – – + – + – dw –
D-Ribose – w + v + d/w – – d – + – – –
Ethanol – – + v – – + – – – w w + N
Ribitol – w w d + + – v d/w – + d/w d/w N
D-Mannitol + + + + + w s + w + + + + +/w
D-Glucitol + + + + w d + –/s d – + d/w d –
DL-Lactate – w – – – – – – – – + + d/w +/s
Succinate d + + + – + + + + + + + + +
Citrate – + + + d – + + d/w + + – + –
D-Gluconate – + – d/– – – – – – – – – – +
Growth tests
Growth in vitamin-free medium w w w –/+ + + – v + – + + + –
Growth at 30 °C – – – – + – – – – – – + + +
Growth at 35 °C – – – – – – – – – – – – – +

Note. 1, T. foliicola; 2, T. carpini; 3, T. virginica; 4, T. inositophila; 5, T. fujianensis; 6, T. guizhouensis; 7, T. kurtzmanii; 8, T. caerulescens; 9, T. aspidistrae; 10, T. arrabidae; 11, T. cunninghamiae; 12, Sr. musae; 13, Sr. ovata; 14, Sa. phatthalungensis. +, positive reaction; –, negative reaction; d, delayed positive; n, not available; s, slowly positive; v, variable; w, weakly positive. All data from this study, except* which were obtained from the original description (Fonseca and Rodrigues 2011; Inácio et al. 2004; Nutaratat et al. 2022).

 Typus.

CHINA • Guizhou Prov.: Guiyang City, Guiyang Botanical Garden, on the phylloplane of Aspidistra elatior, Oct. 2024, D. Lu, NYNU 2411134 (holotype GDMCC 2.541T preserved as a metabolically inactive state, culture ex-type JCM 38266).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is yellowish-cream, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ellipsoidal and cylindrical, 2.3–3.6 × 3.8–9.4 μm, occurring singly or with buds. After 1 month at 20 °C, a pellicle and a sediment are present. In Dalmau plate culture on CMA, simple pseudohyphae are formed. Ascospores formation is not observed on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, inulin, sucrose, melezitose (weak), methyl-α-D-glucoside (delayed and weak), cellobiose (weak), salicin (weak), D-xylose, L-arabinose (delayed and weak), D-arabinose, D-ribose (delayed), glycerol, ribitol (delayed and weak), D-mannitol (weak), D-glucitol (delayed), succinate, and citrate (delayed and weak). No growth occurred in raffinose, melibiose, galactose, lactose, trehalose, maltose, L-sorbose, L-rhamnose, 5-keto-D-gluconate, methanol, ethanol, erythritol, galactitol, myo-inositol, DL-lactate, D-gluconate, D-glucosamine, N-acetyl-D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1,5-lactone. Nitrate, nitrite (weak), and ethylamine (delayed and weak) are utilized as a nitrogen source. L-Lysine and cadaverine are not utilized. Growth in vitamin-free medium is positive. No growth occurred with 1% acetic acid, 0.01% cycloheximide or 10% NaCl plus 5% glucose. Maximum growth temperature is 25 °C. Starch-like polysaccharide is produced. Urease reaction is positive. Diazonium blue B reaction is negative.

 GenBank accession numbers.

Holotype GDMCC 2.541T (ITS: PV839642, D1/D2: PV839641, mtSSU: PX939148).

 Notes.

An unpublished strain, JS-42 isolated from Korea, had identical ITS sequences and similar LSU sequences (with a three nucleotides difference) with T. aspidistrae, which indicated that this new species may be also found in other locations than China.

 Fungi Haplotaxida Taphrinaceae 5C5DA1AF-F406-5E8E-BF05-4F7DAA702B35 Taphrina cunninghamiae 861666 C.Y. Chai & F.L. Hui sp. nov. Fig. 2C, D Etymology.

The specific epithet cunninghamiae refers to Cunninghamia, the plant genus from which the type strain was isolated.

 Diagnosis.

Multi-gene phylogenetic analysis revealed that T. cunninghamiae formed a distinct lineage closely related to T. arrabidae and T. aspidistrae (Fig. 1). T. cunninghamiae differed from T. arrabidae and T. aspidistrae by 14–15 nucleotide substitutions (2.4–2.6%) in the LSU region and 57–67 nucleotide mismatches (9–10.6%) in the ITS region. Physiologically, T. cunninghamiae can be distinguished from T. arrabidae and T. aspidistrae in its ability to assimilate trehalose, maltose, L-rhamnose, ethanol, and DL-lactate (Table 2; Suppl. material 2: table SS2).

 Typus.

CHINA • Guizhou Prov.: Pingtang County, Sifangjing Village, on the phylloplane of Cunninghamia lanceolata, Feb. 2023, D. Lu, NYNU 232134 (holotype GDMCC 2.492T preserved as a metabolically inactive state, culture ex-type PYCC 9978).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is yellowish-cream, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ellipsoidal and cylindrical, 2.2–4.4 × 4.3–6.6 μm, occurring singly or with buds. After 1 month at 20 °C, a pellicle and a sediment are present. In Dalmau plate culture on CMA, pseudohyphae are not formed. Ascospores formation is not observed in any of the strains or when strains are paired on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, inulin, sucrose, trehalose, maltose, melezitose, methyl-α-D-glucoside, cellobiose, salicin, L-rhamnose, D-xylose, L-arabinose, D-arabinose, D-ribose, ethanol (weak), glycerol, ribitol, D-mannitol, D-glucitol, DL-lactate, succinate, citrate, and N-acetyl-D-glucosamine (weak). No growth occurred in raffinose, melibiose, galactose, lactose, L-sorbose, 5-keto-D-gluconate, methanol, erythritol, galactitol, myo-inositol, D-gluconate, D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1,5-lactone. Nitrite, nitrate, and L-lysine are utilized as a nitrogen source. Ethylamine and cadaverine are not utilized. Growth in vitamin-free medium is positive. No growth occurred with 0.01% cycloheximide or 10% NaCl plus 5% glucose. Maximum growth temperature is 25 °C. Starch-like polysaccharide is produced. Urease reaction is positive. Diazonium blue B reaction is negative.

 Additional strain examined.

CHINA • Guizhou Prov.: Pingtang County, Sifangjing Village, on the phylloplane of Cunninghamia lanceolata, Feb. 2023, D. Lu, NYNU 232156.

 GenBank accession numbers.

Holotype GDMCC 2.492T (ITS: OQ851904, D1/D2: OQ851903, mtSSU: PX939040); additional strains NYNU 232156 (ITS: PX070522, D1/D2: PX070523, mtSSU: PX939039).

 Fungi Haplotaxida Taphrinaceae 58C4FC54-7C00-5B72-82E9-6C0908FFBC6B Taphrina fujianensis 861667 C.Y. Chai & F.L. Hui sp. nov. Fig. 2E, F Etymology.

The specific epithet fujianensis refers to the geographic origin of the type strain, Quanzhou city, Fujian Province.

 Diagnosis.

Multi-gene phylogenetic tree showed that T. fujianensis formed a distinct lineage and was sister to T. guizhouensis (Fig. 1). T. fujianensis differed from T. guizhouensis by nine nucleotide substitutions (1.6%) and 40 nucleotide mismatches (6.6%) in the LSU and ITS regions, respectively. Physiologically, T. fujianensis can be differentiated from T. guizhouensis in its inability to assimilate succinate and its ability to assimilate citrate and grow at 30 °C (Table 2; Suppl. material 2: table S3).

 Typus.

CHINA • Fujian Prov.: Quanzhou City, Qingyuan Mountain, on the phylloplane of Dicranopteris sp., Mar. 2022, W.T. Hu, NYNU 224115 (holotype GDMCC 2.310T preserved as a metabolically inactive state, culture ex-type PYCC 9943).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is pinkish-cream, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ovoid, ellipsoidal and cylindrical, 1.9–3.1 × 3.6–5.9 μm, occurring singly or with buds. After 1 month at 20 °C, a pellicle and a sediment are present. In Dalmau plate culture on CMA, pseudohyphae and hyphae are not formed. Ascospores formation is not observed on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, inulin, sucrose, trehalose, maltose, melezitose, methyl-α-D-glucoside, cellobiose, salicin, L-rhamnose (weak), D-xylose, L-arabinose, D-arabinose, D-ribose, glycerol, ribitol, D-mannitol, D-glucitol (weak), citrate (delayed), and N-acetyl-D-glucosamine (weak). No growth occurred in raffinose, melibiose, galactose, lactose, L-sorbose, 5-keto-D-gluconate, methanol, ethanol, erythritol, galactitol, myo-inositol, DL-lactate, succinate, D-gluconate, D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1,5-lactone. Nitrite, nitrate (weak), and L-lysine are utilized as a nitrogen source. Ethylamine and cadaverine are not utilized. Growth in vitamin-free medium is positive. No growth occurred with 1% acetic acid, 0.01% cycloheximide or 10% NaCl plus 5% glucose. Maximum growth temperature is 30 °C. Starch-like polysaccharide is produced. Urease reaction is positive. Diazonium blue B reaction is negative.

 GenBank accession numbers.

Holotype GDMCC 2.310T (ITS: OP278686, D1/D2: OP278685, mtSSU: PX939041).

 Notes.

Another Chinese strain CGMCC 2.5682 shared an identical LSU sequence and a similar ITS sequence (with only two nucleotides differences) with T. fujianensis, indicating that this new species has a wide geographical distribution.

 Fungi Haplotaxida Taphrinaceae E83B5BA5-F229-5E09-A0CE-54971B0D55FC Taphrina guizhouensis 861668 C.Y. Chai & F.L. Hui sp. nov. Fig. 2G, H Etymology.

The specific epithet guizhouensis refers to the geographic origin of the type strain, Pingtang County, Guizhou Province.

 Diagnosis.

The new species T. guizhouensis formed a distinct lineage, closely related to T. fujianensis (Fig. 1). T. guizhouensis differed from T. fujianensis by more than nine nucleotide substitutions (1.6%) and 40 nucleotide mismatches (6.6%) in the LSU and ITS regions, respectively (Suppl. material 2: table S4). Physiologically, T. guizhouensis can be distinguished from T. fujianensis in its ability to assimilate succinate and its inability to assimilate citrate and grow at 30 °C (Table 2; Suppl. material 2: table S4).

 Typus.

CHINA • Guizhou Prov.: Pingtang County, on the phylloplane of Cunninghamia lanceolata, Mar. 2024, D. Lu, NYNU 24380 (holotype GDMCC 2.543T preserved as a metabolically inactive state, culture ex-type JCM 38268).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is yellowish-cream, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ovoid, ellipsoidal and cylindrical, 1.7–3.1 × 3.5–8.8 μm, occurring singly or with buds. After 1 month at 20 °C, a pellicle and a sediment are present. In Dalmau plate culture on CMA, pseudohyphae and hyphae are not formed. Ascospores formation is not observed in any of the strains or when strains are paired on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, inulin, sucrose, trehalose (weak), maltose (delayed), melezitose (weak), methyl-α-D-glucoside (delayed), cellobiose (delayed), salicin (weak), D-xylose (weak), L-arabinose (delayed and weak), D-arabinose, D-ribose (delayed and weak), glycerol, ribitol, D-mannitol (weak), D-glucitol (delayed), and succinate. No growth occurred in raffinose, melibiose, galactose, lactose, L-sorbose, L-rhamnose, 5-keto-D-gluconate, methanol, ethanol, erythritol, galactitol, myo-inositol, DL-lactate, citrate, D-gluconate, D-glucosamine, N-acetyl-D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1, 5-lactone. Nitrate, nitrite (weak), and ethylamine (delayed and weak) are utilized as a nitrogen source. L-Lysine and cadaverine are not utilized. Growth in vitamin-free medium is positive. No growth occurred with 1% acetic acid, 0.01% cycloheximide or 10% NaCl plus 5% glucose. Maximum growth temperature is 25 °C. Starch-like polysaccharide is produced. Urease reaction is positive. Diazonium blue B reaction is negative.

 Additional strain examined.

CHINA • Guizhou Prov.: Pingtang County, Sifangjing Village, on the phylloplane of Rhododendron simsii, Mar. 2024, D. Lu, NYNU 243137.

 GenBank accession numbers.

Holotype GDMCC 2.543T (ITS: PV839639, D1/D2: PV839638, mtSSU: PX939035); additional strains NYNU 243137 (ITS: PX070534, D1/D2: PX070535, mtSSU: PX939144).

 Fungi Haplotaxida Taphrinaceae EF37AABB-EC93-59C2-A4C3-2CC2D3DFA06C Taphrina foliicola 861669 C.Y. Chai & F.L. Hui sp. nov. Fig. 2I, J Etymology.

The specific epithet foliicola refers to the ex-type strain isolated from a leaf.

 Diagnosis.

Based on multi-gene phylogenetic analysis, T. foliicola formed a clade clustered with T. carpini, T. inositophila, and T. virginica with significant support values (Fig. 1). T. foliicola differed from those three known species in this clade by more than three nucleotide substitutions (0.5%) in the LSU region and over 35 nucleotide mismatches (5.9%) in the ITS region (Suppl. material 2: table S5). Physiologically, T. foliicola can be distinguished from T. carpini, T. inositophila, and T. virginica in its inability to assimilate D-arabinose, ribitol, and citrate (Table 2; Suppl. material 2: table S5).

 Typus.

CHINA • Henan Prov.: Neixiang County, Baotianman Nature Reserve, on the phylloplane of Carpinus turczaninowii, Sep. 2022, J.Z. Li, NYNU 2211137 (holotype GDMCC 2.330T preserved as a metabolically inactive state, culture ex-type PYCC 9942).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is yellowish-cream, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ellipsoidal and cylindrical, 3.1–3.9 × 5.6–8.2 μm, occurring singly or with buds. After 1 month at 20 °C, a ring and a sediment are present. In Dalmau plate culture on CMA, pseudohyphae and hyphae are not formed. Ascospores formation is not observed in any of the strains or when strains are paired on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, inulin (delayed), sucrose, raffinose (delayed and weak), trehalose, maltose, melezitose, cellobiose, salicin (delayed and weak), D-xylose, glycerol, D-mannitol, D-glucitol, succinate (delayed), and N-acetyl-D-glucosamine. No growth occurred in melibiose, galactose, lactose, methyl-α-D-glucoside, L-sorbose, L-rhamnose, L-arabinose, D-arabinose, 5-keto-D-gluconate, D-ribose, methanol, ethanol, erythritol, ribitol, galactitol, myo-inositol, DL-lactate, citrate, D-gluconate, D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1, 5-lactone. Nitrite, nitrate (delayed), and L-lysine are utilized as a nitrogen source. Ethylamine and cadaverine are not utilized. Growth in vitamin-free medium is positive. No growth occurred with 1% acetic acid, 0.01% cycloheximide or 10% NaCl plus 5% glucose. Maximum growth temperature is 25 °C. Starch-like polysaccharide is produced. Urease reaction is positive. Diazonium blue B reaction is negative.

 Additional strain examined.

CHINA • Henan Prov.: Neixiang County, Baotianman Nature Reserve, on the phylloplane of Carpinus turczaninowii, Sep. 2022, J.Z. Li, NYNU 221115; Henan Prov.: Songxian County, Tianchi mountain, on the phylloplane of Carpinus turczaninowii, Jun. 2023, J.Z. Li, NYNU 236258; Henan Prov.: Songxian County, Tianchi mountain, on the phylloplane of Cornus officinalis, Jun. 2023, J.Z. Li, NYNU 236334.

 GenBank accession numbers.

Holotype GDMCC 2.330T (ITS: OP959653, D1/D2: OP959502, mtSSU: PX939147); additional strains NYNU 221115 (ITS: OP954718, D1/D2: OP954686, mtSSU: PX939038), NYNU 236258 (ITS: PX063794, D1/D2: PX070529, mtSSU: PX939042), and NYNU 236334 (ITS: PX063790, D1/D2: PX070528, mtSSU: PX939043).

 Notes.

An extra Chinese strain CGMCC 2.2537 had identical LSU sequences and a similar ITS sequence (with two nucleotide difference) with T. foliicola, which indicated that this new species has a wide geographical distribution.

 Fungi C4E8426A-A58E-5F57-84E0-451C9D4A3B83 Srisukozyma 861671 C.Y. Chai & F.L. Hui gen. nov. Type species.

Srisukozyma musae.

 Etymology.

The genus is named in honor of Nantana Srisuk for his contributions to yeast taxonomy.

 Description.

Colonies are yellowish-cream and butyrous. Budding cells are present, and yeast cells divide by multilateral budding. Neither pseudohyphae nor hyphae are produced. Ascospores have not been observed in either individual or mixed cultures. Glucose is not fermented. Diazonium Blue B reaction and urease activity are negative.

 Classification.

Taphrinales, Taphrinomycetes, Taphrinomycotina, Ascomycota.

 Notes.

The new genus Srisukozyma is proposed for the branch represented by two groups NYNU 24437 and NYNU 2411186, which was closely related to Savitreella. The genus is mainly circumscribed by the phylogenetic analysis of the combined ITS, LSU, and mtSSU dataset, in which it occurred as a separate branch within the Taphrinales. The genus Srisukozyma can also be differentiated from its closely related genus Savitreella based on physiological characteristics (Table 2). Srisukozyma species do not assimilate cellobiose, or D-gluconate, whereas species of Savitreella are capable of utilizing these substrates. Moreover, Srisukozyma species are able to assimilate raffinose, a trait not shared by species of Savitreella.

 Fungi 91DA7889-C9F2-5FD7-828A-04AB06FE9A6D Srisukozyma musae 861672 C.Y. Chai & F.L. Hui sp. nov. Fig. 3A, B Etymology.

The specific epithet musae refers to Musa, the plant genus from which the type strain was isolated.

10.3897/imafungus.17.187200.figure3 488F4B3A-9999-5973-9FC0-5D37E447601F

The streak culture grown on YM agar for 7 d at 20 °C and vegetative cells grown in YM broth for 3 d at 20 °C. A, B. Sr. musae NYNU 24437. C, D. Sr. ovata NYNU 2411186. Scale bars: 10 μm.

https://binary.pensoft.net/fig/1753479 Diagnosis.

Multi-gene phylogenetic analysis revealed that Sr. musae was closely related to Sr. ovata (Fig. 1). Sr. musae differed from the closely related species Sr. ovata by 33 nucleotide substitutions (5.7%) in the LSU region and by 88 nucleotide mismatches (13.9%) in the ITS region (Suppl. material 2: table S6). Physiologically, Sr. musae can be differentiated from its closely related species Sr. ovata in its inability to assimilate inulin, methyl-α-D-glucoside, L-arabinose, D-arabinose, and citrate (Table 2; Suppl. material 2: table S6).

 Typus.

CHINA• Hainan Prov.: Sanya City, Madan Village, on the phylloplane of Musa nana, Mar. 2024, S.L. Lv, NYNU 24437 (holotype CICC 33645T preserved as a metabolically inactive state, culture ex-type PYCC 10061).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is yellowish-cream, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ovoid and ellipsoidal, 2.9–4.3 × 4.0–9.2 μm, occurring singly or with buds. After 1 month at 20 °C, a ring and a sediment are present. In Dalmau plate culture on CMA, pseudohyphae and hyphae are not formed. Ascospores formation is not observed in any of the strains or when strains are paired on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, sucrose, raffinose, trehalose, maltose (delayed and weak), melezitose, salicin (delayed and weak), L-sorbose (delayed), D-xylose, ethanol (weak), glycerol, ribitol (delayed and weak), D-mannitol, D-glucitol (delayed and weak), DL-lactate, and succinate. No growth occurred in inulin, melibiose, galactose, lactose, methyl-α-D-glucoside, cellobiose, L-rhamnose, L-arabinose, D-arabinose, 5-keto-D-gluconate, D-ribose, methanol, erythritol, galactitol, myo-inositol, citrate, D-gluconate, D-glucosamine, N-acetyl-D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1,5-lactone. Nitrite (delayed and weak), nitrate (delayed and weak), and L-lysine (delayed) are utilized as a nitrogen source. Ethylamine and cadaverine are not utilized. Growth in vitamin-free medium is positive. No growth occurred with 0.01% cycloheximide or 10% NaCl plus 5% glucose. Maximum growth temperature is 30 °C. Diazonium Blue B reaction and urease activity are negative.

 Additional strain examined.

CHINA• Hainan Prov.: Sanya City, Madan Village, on the phylloplane of Musa nana, Mar. 2024, S.L. Lv, NYNU 244126; Hainan Prov.: Sanya City, Baogu Village, on the phylloplane of Musa nana, Mar. 2024, S.L. Lv, NYNU 244111; Hainan Prov.: Sanya City, Lanji Village, on the phylloplane of Musa nana, Mar. 2024, S.L. Lv, NYNU 24449.

 GenBank accession numbers.

Holotype CICC 33645T (ITS: PP849688, D1/D2: PP837701, mtSSU: PX939036); additional strains NYNU 244126 (ITS: PX070522, D1/D2: PX070523, mtSSU: PX939145), NYNU 244111 (ITS: PX070525, D1/D2: PX070524, mtSSU: PX939143), and NYNU 24449 (ITS: PX070527, D1/D2: PX070526, mtSSU: PX939037).

 Fungi E666146C-7F76-5205-987D-7B8575B78DC8 Srisukozyma ovata 861674 C.Y. Chai & F.L. Hui sp. nov. Fig. 3C, D Etymology.

The specific epithet ovata refers to the ovoid cell morphology of the type strain.

 Diagnosis.

In the multi-gene phylogenetic tree, Sr. ovata clustered with Sr. musae with high support values (Fig. 1). Sr. ovata differed from the closely related species Sr. musae by 33 nucleotide substitutions (5.7%) in the LSU region and by 88 nucleotide mismatches (13.9%) in the ITS region (Suppl. material 2: table S7). Physiologically, Sr. ovata can be differentiated from the closely related species Sr. musae in its ability to assimilate inulin, methyl-α-D-glucoside, L-arabinose, D-arabinose, and citrate and its inability to assimilate L-sorbose (Table 2; Suppl. material 2: table S7).

 Typus.

CHINA• Guizhou Prov.: Guiyang City, Guiyang Botanical Garden, on the phylloplane of Polygonum chinense, Oct. 2024, D. Lu, NYNU 2411186 (holotype GDMCC 2.542T preserved as a metabolically inactive state, culture ex-type JCM 38267).

 Description.

On YM agar after 7 days at 20 °C, the streak culture is orange-yellow, butyrous, and smooth, with an entire margin. In YM broth after 3 days at 20 °C, the cells are ovoid and ellipsoidal, 2.3–2.8 × 3.3–6.4 μm, occurring singly or with buds. After 1 month at 20 °C, a pellicle and a sediment are present. In Dalmau plate culture on CMA, pseudohyphae and hyphae are not formed. Ascospores formation is not observed in any of the strains or when strains are paired on CMA, 5% ME agar, Fowell’s acetate agar, Gorodkowa agar or V8 agar. Fermentation is absent. The following carbon compounds are assimilated in liquid medium: glucose, inulin, sucrose, raffinose (delayed), trehalose, maltose, melezitose, methyl-α-D-glucoside (delayed and weak), salicin (delayed and weak), D-xylose (delayed), L-arabinose (delayed and weak), D-arabinose (delayed and weak), ethanol, glycerol, ribitol (delayed and weak), D-mannitol, D-glucitol (delayed), DL-lactate (delayed and weak), succinate, and citrate. No growth occurred in melibiose, galactose, lactose, cellobiose, L-sorbose, L-rhamnose, 5-keto-D-gluconate, D-ribose, methanol, erythritol, galactitol, myo-inositol, D-gluconate, D-glucosamine, 2-keto-D-gluconate, D-glucuronate, and glucono-1, 5-lactone. Nitrite (delayed and weak), nitrate (delayed and weak), ethylamine (delayed and weak), and L-lysine are utilized as a nitrogen source. Cadaverine is not utilized. Growth in vitamin-free medium is positive. No growth occurred with 0.01% cycloheximide or 10% NaCl. Maximum growth temperature is 30 °C. Starch-like polysaccharide is produced. Diazonium Blue B reaction and urease activity are negative.

 Additional strain examined.

CHINA • Guizhou Prov.: Guiyang City, Guiyang Botanical Garden, on the phylloplane of Polygonum chinense, Oct. 2024, D. Lu, NYNU 2411221.

 GenBank accession numbers.

Holotype GDMCC 2.542T (ITS: PV843161, D1/D2: PV839685, mtSSU: PX939146); additional strain NYNU 2411221 (ITS: PX070537, D1/D2: PX070536, mtSSU: PX939149).

 Discussion

Species of the genus Taphrina have traditionally been recognized based on their host range, geographical distribution, morphology of sexual forms, and their effects on host plant tissues (Mix 1949; Petrýdesová et al. 2013, 2016). These species are also distinguished by a unique combination of physiological characteristics and the carbohydrate composition of their cell walls (Bacigálová et al. 2003; Petrýdesová et al. 2016). However, Taphrina may not be strictly associated with specific host plants (Mix 1949; Fonseca and Rodrigues 2011), and some species without sexual states lack clear associations with particular host plants (Inácio et al. 2004; Petrýdesová et al. 2016). The application of molecular tools for species identification has facilitated the connection between sexual and asexual morphs, even when the two forms are morphologically dissimilar (Selbmann et al. 2014). Following the polyphasic approach recommended by Selbmann et al. (2014) and Petrýdesová et al. (2016), the present study identified five new Taphrina species and one new genus Srisukozyma with two new species, all of which were observed only in their yeast states.

To date, 43 species are phylogenetically recognized in Taphrina, including T. aspidistrae, T. cunninghamiae, T. foliicola, T. fujianensis, and T. guizhouensis from the present study (Petrýdesová et al. 2016). Among the species of the genus Taphrina, at least four, T. caerulescens, T. populina, T. robinsoniana, T. wiesneri, exhibit high intraspecific sequence variation (Rodrigues and Fonseca 2003). For example, three strains of T. populina had similar LSU sequences (with a three nucleotides difference), but possessed 13–14 nucleotide differences (2.1–2.2%) in the ITS region. In the case of T. wiesneri, four strains belong to this species differed from each other by one nucleotide substitutions in the LSU region and 1–14 nucleotide mismatches (0.2–2.3%) in the ITS region. The result of sequence similarity analysis indicated that members of the genus Taphrina exhibit high intraspecific sequence variation, but high intraspecific sequence variation was found only in the ITS region, not in the LSU region (Rodrigues and Fonseca 2003). In agreement with previous observation, intraspecific variability was also reported for mating strains (CBS 490 and CBS 2630) of the heterothallic species Sporidiobolus salmonicolor, which were found to differ from one another by seven nucleotide substitutions (1.2%) and 26 nucleotide mismatches (4.5%) in the LSU and ITS regions, respectively (Dayo-Owoyemi et al. 2013). Although the ITS region was selected as a universal DNA-barcode for fungal classification, primarily due to its high discriminative power (Schoch et al. 2012), it is clear from this study that this region alone is not sufficient for species delimitation in yeast species, especially within the Taphrina clade. Thus, it is important to keep in mind that both gene regions are necessary for a reliable species delimitation in the Taphrina clade.

Two new Srisukozyma species clustered in the clade as Savitreella with strong support. However, low sequence similarity was observed between Srisukozyma species and any known taxa. The phylogenetic analysis and the comparison of predicted taxonomic thresholds demonstrated that Srisukozyma represents a new genus within the Taphrinales. The new genus is also physiologically distinct from other genera within Taphrinales. For instance, Srisukozyma species do not assimilate cellobiose, whereas species of the genera Savitreella and Taphrina are capable of utilizing these substrates. Moreover, Srisukozyma species are able to assimilate raffinose, a trait not shared by species of Savitreella. Since the family assignment of the Taphrinomycotina is still unclear, we chose not to assign the family of the new genus and have left open the family assignment until the existing family structure within the Taphrinomycotina is better clarified, based on eventual phylogenomic data. Although the three-locus dataset (ITS, LSU, mtSSU) used in this study has inherent limitations, this marker combination is the current standard for species-level taxonomy in Taphrinales. Together with extensive physiological and morphological data, it provides robust support for the new taxa described herein. Future phylogenomic analyses based on whole-genome sequencing, even at low coverage, will be necessary to resolve deeper nodes within Taphrinomycotina and to refine family-level classifications.

Members of Taphrina are widely distributed and inhabit various environments. The sexual, filamentous state exclusively exists as a biotrophic plant pathogen on Rosaceae, Betulaceae, and Fagaceae (Selbmann et al. 2014). The asexual yeast states exhibit saprobic growth and have been isolated from a variety of substrates, including flowers (Golonka and Vilgalys 2013), leaves (Inácio et al. 2004; Petrýdesová et al. 2016), leaf litter (Babjeva and Reshetova 1998; Maksimova and Chernov 2004; Yurkov et al. 2004), forest soil (Wuczkowski et al. 2005), and even Antarctic rock (Selbmann et al. 2014). Furthermore, ITS sequences matching various Taphrina species were identified among endophytic fungi colonizing bur oak (Quercus macrocarpa) in the Flint Hills region, USA (Jumpponen and Jones 2009, 2010), and beech (Fagus sylvatica) in the French Pyrenees (Cordier et al. 2012). Among all the reported Taphrina species, most are associated with plant leaves. The five new species described in this paper were also isolated from leaf surfaces, further enriching the diversity of phylloplane fungi. Additionally, members of Taphrina are generally recognized as phytopathogens. Many Taphrina species are plant parasites that cause gall formation on stems, leaf curls, and witches’ brooms, particularly on economically important fruit trees such as peach, plum, and cherry (Selbmann et al. 2014). Consequently, the growing interest in these fungi extends beyond their taxonomy and ecological roles to include their potential pathogenicity.

 Conclusion

This study reveals a new monotypic genus Srisukozyma, comprising two new species Sr. musae and Sr. ovata, along with five additional new Taphrina species: T. aspidistrae, T. cunninghamiae, T. foliicola, T. fujianensis, and T. guizhouensis, all isolated from leaf surfaces at different locations across China. Phylogenetic analyses and phenotypic characterization support the seven new taxa as members of Taphrinales (Fig. 1). Detailed descriptions, illustrations, and comparative discussions with their nearest relatives are provided. This study represents the first comprehensive analysis of Taphrinales species associated with plant materials from tropical and subtropical China. The findings expand our understanding of generic and species diversity within this order. We anticipate that future research will uncover more species and genera within the order.

 Acknowledgements

We extend our sincere gratitude to Lu Dan and Shi-Long Lv for their invaluable assistance in collecting samples from the Guizhou and Hainan provinces. We are also grateful to Dr. José Paulo Sampaio of the Portuguese Yeast Culture Collection (PYCC) for his generous assistance with sequencing the type strains.

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The authors have declared that no competing interests exist.

 Ethical statement

No ethical statement was reported.

 Artificial Intelligence (AI) use

The authors accept full responsibility for the content of the manuscript, including the disclosure of any use of AI.

No AI tools were used in the preparation of this manuscript.

 Adherence to national and international regulations

All the fungal strains used in this study have been legally obtained, respecting the Convention on Biological Diversity (Rio Convention).

 Funding

This research was funded by Henan Provincial Key Scientific and Technological Research (Project No. 262102110018), the National Natural Science Foundation of China (Project No. 31570021), and Agricultural Biomass Green Conversion Technology University Scientific Innovation Team in Henan Province, China (Project No. 24IRTSTHN036).

 Author contributions

Chun-Yue Chai: investigation, methodology, visualization, data curation, and writing—original draft; Dan-Yang Cai, Bing-Yan Song, and Rui-Xiu Wang: investigation; Feng-Li Hui: conceptualization, methodology, funding acquisition, and writing—review and editing.

 Author ORCIDs

C.-Y. Chai https://orcid.org/0000-0001-7753-6223

D.-Y. Cai https://orcid.org/0009-0002-6866-7322

B.-Y. Song https://orcid.org/0009-0000-6802-5077

R.-X. Wang https://orcid.org/0009-0009-5141-7096

F.-L. Hui https://orcid.org/0000-0001-7928-3055

 Data availability

All gene sequences generated in this study are available in GenBank under the accession numbers provided, with all additional data included in the manuscript and its supplements.

 Supplementary materials 10.3897/imafungus.17.187200.suppl1 791ACF77-5024-587A-AEE9-CBD20B28EBE0

A dataset of the combined ITS, LSU, and mtSSU sequences for Fig. 1

fas

The concatenated dataset of the three regions had a total length of 1,979 bp, consisting of 598 bp for the ITS region, 574 bp for the LSU region, and 807 bp for the mtSSU region.

 https://binary.pensoft.net/file/1753480 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. Chun-Yue Chai, Dan-Yang Cai, Bing-Yan Song, Rui-Xiu Wang, Feng-Li Hui 10.3897/imafungus.17.187200.suppl2 AD82AE2B-168C-5176-A063-208E8046BAA5

Supplementary table SS1–S7

xlsx

Physiological characteristics and nucleotide divergences that differentiate new species from their closest species. Each new species provided as a separate Excel sheet.

 https://binary.pensoft.net/file/1753481 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. Chun-Yue Chai, Dan-Yang Cai, Bing-Yan Song, Rui-Xiu Wang, Feng-Li Hui