Mycol Progress DOI 10.1007/s11557-017-1276-2

ORIGINAL ARTICLE

Taxonomic revisions in the Microstromatales: two new yeast species, two new genera, and validation of Jaminaea and two Sympodiomycopsis species Teeratas Kijpornyongpan 1 & M. Catherine Aime 1

Received: 12 February 2016 / Revised: 24 January 2017 / Accepted: 30 January 2017 # German Mycological Society and Springer-Verlag Berlin Heidelberg 2017

Abstract Environmental sampling yielded two yeast species belonging to Microstromatales (Exobasidiomycetes, Ustilaginomycotina). The first species was collected from a leaf phylloplane infected by the rust fungus Coleosporium plumeriae, and represents a new species in the genus Jaminaea, for which the name Jaminaea rosea sp. nov. is proposed. The second species was isolated from air on 50% glucose media and is most similar to Microstroma phylloplanum. However, our phylogenetic analyses reveal that species currently placed in Microstroma are not monophyletic, and M. phylloplanum, M. juglandis and M. albiziae are not related to the type species of this genus, M. album. Thus, Pseudomicrostroma gen. nov. is proposed to accommodate the following species: P. glucosiphilum sp. nov., P. phylloplanum comb. nov. and P. juglandis comb. nov. We also propose Parajaminaea gen. nov. to accommodate P. albizii comb. nov. and P. phylloscopi sp. nov. based on phylogenetic analyses that show these are not congeneric with Jaminaea or Microstroma. In addition, we validate the genus Jaminaea, its respective species and two species of Sympodiomycopsis and provide a new combination, Microstroma bacarum comb. nov., for the anamorphic yeast Rhodotorula bacarum. Our results illustrate non-monophyly of Quambalariaceae and Microstromataceae as currently Section Editor: Dominik Begerow Electronic supplementary material The online version of this article (doi:10.1007/s11557-017-1276-2) contains supplementary material, which is available to authorized users. * M. Catherine Aime [email protected] 1

Department of Botany and Plant Pathology, Purdue University, 915 W State St, West Lafayette, IN 47907-2054, USA

circumscribed. Taxonomy of Microstroma and the Microstromataceae is reviewed and discussed. Finally, analyses of all available small subunit rDNA sequences for Jaminaea species show that J. angkorensis is the only known species that possess a group I intron in this locus, once considered a potential feature indicating the basal placement of this genus in Microstromatales. Keyword Basidiomycetous yeast . Exobasidiomycetes . Smut fungi . rDNA sequences . Taxonomic validation

Introduction Ustilaginomycotina is a diverse subphylum of fungi in Basidiomycota containing approximately 1700 species belonging to 15 orders (Begerow et al. 2014; Albu et al. 2015; Wang et al. 2015; Riess et al. 2016). Most known members are Bsmut^ fungi, which are dimorphic biotrophic plant pathogens, the majority of which belong to Ustilaginomycetes. However, recent molecular phylogenetic studies have expanded Ustilaginomycotina to include many fungi for which sexual or infectious stages are unknown. These may be animalassociated fungi, potential phytopathogens, or nonpathogenic filamentous fungi and yeasts known from soils, phylloplanes or environmental sampling (Begerow et al. 2014). Microstromatales is one of the orders in Exobasidiomycetes, the second major class of Ustilaginomycotina. It was first recognized as a smut-allied group of obligate phytopathogens that do not produce teliospores and consisted of only the genus Microstroma when it was established (Bauer et al. 1997). Subsequent classification of Microstromatales has mostly relied on molecular phylogenetic studies, resulting in inclusion of three anamorphic yeast and yeast-like genera—Jaminaea,

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Quambalaria and Sympodiomycopsis—and three species of BRhodotorula^ (Begerow et al. 2001; de Beer et al. 2006; Sipiczki and Kajdacsi, 2009). Rhodotorula bacarum is now considered an anamorph of M. album, and R. hinnulea and R. phylloplana have been combined into a single species, Microstroma phylloplanum (Sampaio 2011; Wang et al. 2015). The order currently contains more than 20 described species in six genera (Begerow et al. 2014; Chen et al. 2013; Francesca et al. 2016; Wang et al. 2015) which have been distributed in three families, Microstromataceae, Quambalariaceae, and Volvocisporiaceae (Jülich 1981; Begerow et al. 2001; de Beer et al. 2006; Begerow et al. 2014). Here, we describe two new Microstromatales yeast species based on rDNA sequence data, assimilation profiles and culture studies. Because Microstroma is polyphyletic, we propose two new genera for those species that are not congeneric with M. album, the type. Finally, we validate the genus Jaminaea, its respective species and two Sympodiomycopsis species, and propose several new combinations in order to provide a more natural taxonomic classification for members of this order.

Materials and methods Yeast isolation The yeast MCA4718 was isolated from a private house in Zemmer, Germany (49°53′50″N, 6°42′1″E) in April 2012. A potato-dextrose agar (PDA) plate containing 50% glucose (w/w) was exposed to air indoors overnight, and subsequently incubated at room temperature (25 °C) until colonies were visible. Colonies were isolated with a sterile toothpick and subcultured on fresh PDA plates. A second species, MCA5214, was isolated from the phylloplane of a Plumeria sp. (Apocynaceae) infected with the rust fungus Coleosporium plumeriae and collected from a private garden in Coral Gables, Florida (25°43′34″N, 80°15′47″W) in July 2013 using the spore drop method described in Toome et al. (2013). Cultures were maintained on PDA slants at 4 °C and in 40% glycerol vials at −80 °C for the duration of this study and have been deposited in Centraalbureau voor Schimmelcultures (CBS) Fungal Biodiversity Center and Agricultural Research Service (NRRL) Culture Collection under the collection nos. CBS 14053 and NRRL 66310 for MCA4718, and CBS 14051 and NRRL 66311 for MCA5214. DNA sequencing DNA was extracted for each isolate as follows: the isolate was cultured on PDA and incubated at room temperature (25 °C) for 4 days, after which DNA extraction was completed using a Promega Wizard extraction kit (Promega, Madison, WI,

USA) following the manufacturer’s protocols. Amplification was carried out with the following primers: the large subunit rDNA (LSU) with LR0R/LR6 (Vilgalys and Hester 1990; Vilgalys et al. 1994), the internal transcribed spacer 1–5.8S– internal transcribed spacer 2 (ITS) region with ITS1F/ITS4 (White et al. 1990; Gardes and Bruns 1993) and the small ribosomal subunit rDNA (SSU) with NS1/NS4 and NS3/ NS8 for MCA4718 and PNS1/NS6 and NS5/NS8 for MCA5214 (White et al. 1990; Hibbett 1996). Amplification conditions for the LSU region started with 95 °C for 5 min followed by 35 cycles of 94 °C for 30 s, 50 °C for 45 s and 72 °C for 1 min, ending with 72 °C for 7 min as a final extension step. The same PCR conditions were used for the ITS and SSU regions, except that the annealing step was set at 58 °C for 30 s for SSU and the extension step was changed to 45 s for ITS and 1.30 min for SSU. All PCR products were evaluated for size and quality in 1% agarose gels. PCR products were sent to Beckmann Coulter (Indianapolis, IN, USA) for DNA sequencing. Each DNA sequence was edited in Sequencher 5.2.3 (Gene Code, Ann Arbor, MI, USA), and then analyzed using BLASTn through the GenBank database (http:// blast.ncbi.nlm.nih.gov/Blast.cgi) under the megablast algorithm for initial identification as members of Microstromatales. To obtain the DNA sequences of RNA polymerase II second largest subunit (RPB2) and translation elongation factor 1 alpha (TEF1) of these two isolates, the RPB2 and TEF1 sequences of Sympodiomycopsis kandeliae (GenBank accessions KP323077 and KP323149, respectively) were used to blast to the reference genomes of the isolate MCA4718 and MCA5214 through JGI MycoCosm portal (Kijpornyongpan and Aime, unpublished; http://jgi.doe.gov/data-and-tools/ mycocosm/). The hit sequences from the reference genomes were downloaded and used for phylogenetic analyses as described below.

Sequence alignment and phylogenetic analyses Fungal species of Microstromatales having ITS, LSU, RPB2 and TEF1 sequences in the NCBI GenBank database were included in the analyses (Table S1). The sequences of each DNA region were separately aligned and manually trimmed using MUSCLE (Edgar 2004) performed in MEGA6 (Tamura et al. 2013). The molecular data were split into three different datasets based on sequence availabilities and sampling levels: (1) LSU dataset, which consisted of the most extensive sampling of Microstromatales taxa, (2) concatenated ITS-LSU data, which consisted of representative species of each genus in Microstromatales, and (3) concatenated ITS-LSU-RPB2TEF1 data, which was used for resolving phylogenetic relationships among genera in the order. Phragmotaenium oryzicola and P. derxii were selected as outgroup taxa based on the study of Wei et al. (2011).

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The optimal nucleotide substitution model for each dataset was determined by built-in function in MEGA6. The neighbor-joining method was used for phylogenetic reconstruction of dataset (1) using MEGA6. Kimura-2-parameter model with substitution-rate among sites of gamma distribution (K2+G) was set as a nucleotide substitution model, and 1,000 pseudoreplicates bootstrapping was used to evaluate a node support. For datasets (2) and (3), maximum-likelihood (ML) and maximum parsimony (MP) tree reconstructions were performed by MEGA6 with 1000 bootstrap pseudoreplicates. General time reversible model with substitution-rate among sites of gamma distribution (GTR+G) and Tamura-Nei model with substitution-rate among sites of gamma distribution with invariant sites (TN93+G+I) were set as nucleotide substitution models for datasets (2) and (3), respectively. Datasets (2) and (3) were also used for Bayesian phylogenetic reconstruction using BEAST package v.1.8.3 (Drummond and Rambaut 2007; Drummond et al. 2012). The substitution models from ML analyses were also applied for the Bayesian inference. Yule speciation process was used as a tree prior model (Gernhard 2008). The Monte Carlo chain was run for 10,000,000 generations, and a tree was sampled every 1,000 generations. Other parameters were set as default. The majority-rule consensus tree was computed by targeting maximum clade credibility, and the first 1,000 sampled trees were discarded as burn-in. Finally, the available SSU sequences of Jaminaea species and other representative Ustilaginomycotina taxa in Microstromatales, Entylomatales, Exobasidiales and Ustilaginales (Table S2) were separately aligned by MUSCLE with default parameters to evaluate for the presence of group I introns.

Morphology and assimilation studies To observe colony morphology, each fungal isolate was cultured on the following media: PDA, corn meal agar (CMA), yeast malt agar (YMA) and yeast peptone glucose agar (YPGA). Morphological characteristics were observed after incubation at room temperature (25 °C) for 14 days. Color was determined using color codes from The Online Auction Color ChartTM (2004). To study microscopic morphology, cells of each isolate were cultured in yeast malt broth (YMB) and incubated at room temperature (25 °C) for 7 days. Cell shape, size and other morphological characters were observed under differential interference contrast microscopy with a ×100 objective lens. Assimilation of various single carbon sources was determined on YT Microplate™ (Biolog, Hayward, CA, USA). Only carbon compounds having positive or negative results from both oxidation test and growth test were recorded. Assimilation tests of other single carbon compounds not present in YT Microplate™, nitrogen assimilation, fermentation

and high osmotic pressure tests were performed according to the method described by Suh et al. (2008).

Results and discussion Species and generic delimitation We sequenced the LSU D1/D2 region, which has been traditionally used as the DNA barcode for yeasts (Kurtzman and Robnett 1998; Fell et al. 2000), and the ITS, which serves as the default barcode for Fungi (Schoch et al. 2012). Blast analyses of the D1/D2 region revealed that both isolates belonged to Microstromatales: MCA4718 was most similar to the extype CBS8073 of Microstroma phylloplanum (accession: AF190004, identity at 529/533 bp with 2 substitutions and 2 indels) and MCA5214 was most similar to the ex-type CBS10858 of Jaminaea lanaiensis (accession: KP322964, identity at 555/558 bp with 1 substitution and 2 indels). Blast analyses of the ITS region concurred with the D1/D2 region: MCA4718 was most similar to the ex-type of M. phylloplanum (accession: AB038131, identity at 678/ 688 bp with 9 substitutions and 1 indel) and MCA5214 was most similar to the ex-type of J. lanaiensis (accession: KP322964, identity at 613/621 bp with 7 substitutions and 1 indel). Phylogenetic analyses of multi-locus datasets for these two, including all closely related species of Microstromatales that have available sequences, were performed in order to refine their taxonomic placements (Figs. 1, S1). To clarify species delimitations of our two isolates, we performed assimilation studies and compared with their closely related described species (Tables 1, 2). The combined data for both isolates show significant differences from their sibling species, indicative of new species. MCA5214 belongs to the genus Jaminaea and represents the first isolate of a species of Jaminaea from a leaf phylloplane infected with a rust fungus. Meanwhile, although MCA4718 appears congeneric with M. phylloplanum and M. juglandis, they are phylogenetically distinct from M. album, the type species of Microstroma (Figs. 1, S2). Niessel first described the genus Microstroma in 1861, with the type species M. album. Microstroma species are obligate plant pathogenic fungi of woody plant leaves that cause characteristic white powdery infections produced by fascicles of basidia that protrude through host stomata (Pires 1928; Cooke 1962). Currently, there are between 4 and 26 accepted species in the genus (Kirk et al. 2008; http://www.indexfungorum. org). However, only a handful of these, e.g., M. album and M. juglandis, have received minimal study and the fact that these species are obligate parasites in nature has rendered them difficult to isolate or study in pure culture (Pires 1928; Cooke 1962; von Arx et al. 1982). Due to a lack of adequate data for most described species, few taxonomic treatments

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Fig. 1 Phylogenetic analyses of rDNA sequences among representative fungal species in Microstromatales. Concatenated sequences of ITS and LSU region were analyzed through ML, MP and Bayesian tree reconstruction. Consensus topology in agreement with at least two different methods of tree reconstruction was shown. Phragmotaenium oryzicola and P. derxii were used as the outgroup. Numbers on branches indicate bootstrap support levels (>50%) from ML and MP analyses and posterior probabilities (>0.7) from Bayesian analyses,

respectively. Nodes having a bullet indicate the strong node support (>50% bootstrap or > 0.7 posterior probability) obtained from four-gene dataset (Fig. S1). Members of each family sensu Begerow et al. (2014) are labeled in color-shaded boxes: pink Microstromataceae; blue Quambalariaceae; green Volvocisporiaceae; white unplaced. Sequences from GenBank are indicated by their accession numbers in parentheses. Superscript T refers to the ex-type of a species. Bar 0.05 substitutions per nucleotide position

exist for this genus, and members have been the target of historical taxonomic disagreement (Pires 1928; Cooke 1962; von Arx et al. 1982). Recent systematic studies on Microstromatales were primarily based on molecular phylogenetics. This resulted in recognition of several yeast and yeast-like species—such as Rhodotorula hinnulea, R. phylloplana, R. bacarum and species of Sympodiomycopsis, Quambalaria and Jaminaea—for which the sexual stages are not yet known (e.g., Begerow et al. 2001; de Beer et al. 2006; Sipiczki and Kajdacsi, 2009). Sampaio (2011) considered R. hinnulea as a synonym of R. phylloplana, and merged these two species into a single taxon, R. phylloplana. Subsequently, Wang et al. (2015) recombined R. phylloplana into Microstroma based on a phylogenetic sister relationship between R. phylloplana, M. juglandis and M. album despite gene tree discordances

across some of the datasets. Based on our rDNA dataset, which provides more robust taxon sampling than that of Wang et al. (2015), Microstroma is not monophyletic and M. juglandis does not appear to be congeneric with the generic type, M. album (Figs. 1, S2). Although rDNA sequence data provide strong support for delimiting genera, they are not robust enough for resolving relationships between genera in Microstromatales. Thus, we include DNA sequences of RPB2 and TEF1, two proteincoding regions, in the four-gene dataset that includes exemplars from all genera-level clades of interest, providing deeper level support for most nodes (Fig. S1). Results are consistent in all analyses (ML, MP, and Bayesian) in showing that M. juglandis and M. phylloplanum do not belong to Microstroma, based on non-monophyly with R. bacarum, which was synonymized with M. album in Wang et al.

Mycol Progress Table 1 Physiological characteristics of Pseudomicrostroma glucosiphilum and closely related species (P. phylloplanum and P. juglandis)

Characteristics

Pseudomicrostroma glucosiphilum MCA4718T

P. phylloplanum

P. juglandis

(Microstroma phylloplanum)

(Microstroma juglandis)

CBS8073T

CBS528.83/ RB2054

C assimilation Cellobiose

+

w

d

Trehalose D-Xylose

+ d

w +

+ −

D-Arabinose

+

−

−

D-Ribose

+

−

+

Ribitol α-Methyl-D-glucoside

− w

− +

ND dw

Salicin DL-Lactate

− −

+ +

− −

Citric acid

+

−

−

D-mannitol Soluble starch N assimilation

+ w

+ w

d −

Nitrite Growth on 50% (w/w) glucose YE ager Growth at 30 °C

+ +

+ −

− ND

+

−

+

All species can assimilate these carbon compounds in common: D-glucose, sucrose, maltose, raffinose, melezitose, L-arabinose, ethanol, glycerol, i-erythritol, i-inositol and D-glucitol. Nitrogen assimilation on nitrate are positive in all species. These three species can grow in vitamin-free medium. Assimilation data of P. phylloplanum are from Shivas and Rodrigues de Miranda (1983). Physiological data of P. juglandis are from Middelhoven et al. (2000) and Sampaio (2011). + growth; − no growth; w weak growth; d delayed growth; ND not determined. T

indicates the ex-type

(2015) (Fig. S1). Because there are no available genera in which to place MCA4718, M. phylloplanum, and M. juglandis we propose a new genus Pseudomicrostroma gen. nov. to accommodate these. Rhodotorula bacarum was treated as a synonym of M. album based solely on LSU sequence data in Wang et al. (2015). However, Sampaio (2011) considered these to be close relatives, but not conspecific, which is in agreement with all of our phylogenetic analyses (Figs. 1, S2). This is supported by culture characteristics given that R. bacarum is only known as a yeast form, whereas the only recorded culture M. album (CBS654.85) is indicated as having filamentous growth. However, given the lack of additional culture data for M. album, additional studies are needed to definitely delimit these taxa. Microstroma albiziae, a foliar pathogen on the woody plant genus Albizia, does not appear to be congeneric with M. album, but is placed as sister to J. phylloscopi (Figs. 1, S2). Jaminaea phylloscopi was originally assigned to the genus Jaminaea based on phylogenetic relatedness, using LSU and ITS sequences, to other described Jaminaea species (Francesca et al. 2016). However, our results suggest that

J. phylloscopi, and M. albiziae, are phylogenetically distinct from Jaminaea (Figs. 1, S2). Jaminaea phylloscopi produces neither pseudohyphae nor true hyphae in culture, which differs from the other described Jaminaea species (Mahdi et al. 2008; Sipiczki and Kajdacsi 2009; Francesca et al. 2016). Based on these data, we propose the new genus Parajaminaea gen. nov. to accommodate M. albiziae and J. phylloscopi. The genus name Jaminaea was first proposed with the type species J. angkorensis, to accommodate a yeast that is phylogenetically distinct from previously described Microstromatales genera (Sipiczki and Kajdacsi, 2009). However, according to the international code of nomenclature for algae, fungi and plants (ICN) (McNeill et al. 2012), the name J. angkorensis was invalidly published as the herbarium/culture collection/institution of the type was not designated (Art. 40.7), and whether/how the type was permanently preserved in metabolically inactive state was not indicated (Art 8.4). Consequently, the genus name Jaminaea was also invalidly published (Art. 40.1 and 40.3). Two subsequently published names in Jaminaea, J. lanaiensis (a new combination) and J. phylloscopi (a new species) (Wei et al. 2011; Francesca

Mycol Progress Table 2 Physiological characteristics of Jaminaea rosea and closely related species

Characteristics

Jaminaea rosea T

MCA5214

J. angkorensis T

CBS10918

J. lanaiensis T

CBS10858

P. phylloscopi (J. phylloscopi) 551B6T

C assimilation D-Galactose

−

+

−

d

D-Ribose D-Arabinose

+ −

+ +

− −

+ d

L-Arabinose L-Rhamnose

+ −

ND d

− −

+ −

Trehalose

−

d

−

+

Salicin Lactose

− +

d −

− −

d +

i-Erythritol i-inositol

+ w

− +

− −

+ −

Ribitol

−

+

−

d

Xylitol D-Galacturonate

w −

− w

ND ND

d −

Sucrose Cellobiose Melibiose

+ + +

+ + +

− − +

+ + −

Melezitose D-Xylose

+ +

+ +

− −

+ +

Ethanol

+

+

−

+

Citrate Soluble starch N assimilation Nitrite (NaNO2) L-Lysine Growth at 37 °C

+ +

+ +

+ +

− w

d − +

− + −

ND ND −

d d +

All species can assimilate these carbon compounds in common: D-glucose, maltose, L-sorbose, glycerol, Dmannitol, succinic acid, raffinose and DL-lactate. Nitrogen assimilation on nitrate are positive in all species. These four species can grow on 50% glucose agar medium. Assimilation data from Jaminaea angkorensis, J. lanaiensis and Parajaminaea phylloscopi are from Sipiczki and Kajdacsi (2009), Mahdi et al. (2008) and Francesca et al. (2016), respectively. + growth; − no growth; w weak growth; d delayed growth; ND not determined. T

indicates the ex-type

et al. 2016), were also invalid due to application of an invalid genus name (Art 35.1). Furthermore, the basionym for J. lanaiensis, Sympodiomycopsis lanaiensis as well as the new name, J. phylloscopi, were also published in violation of Art 8.4 and 40.7 (Mahdi et al. 2008; Francesca et al. 2016). The name S. kandeliae (Wei et al. 2011) was also invalidly published in accordance with Art. 8.4 and 40.7. Finally, even though the description of S. yantaiensis (Chen et al. 2013) indicated lyophilized cultures deposited in two public culture collections (follow Rec. 8B1), the designated holotype NYNU121010 did not include the information of herbarium/ culture collection, and it was not permanently preserved in a metabolically inactive state (resulting in violation of Art 8.4 and 40.7). Taken altogether, this is an exemplar case of how

improper typification of culturable yeasts/fungi can have an adverse effect on fungal taxonomy. Our study shows that none of the families thus far delineated in Microstromatales—Volvocisporiaceae, Quambalariaceae, and Microstromataceae (Jülich 1981; Begerow et al. 2001; de Beer et al. 2006)—are reciprocally monophyletic (Figs. 1, S1, S2). Interestingly, the deepest division in Microstromatales occurs between Jaminaea and the other members of the order, which was earlier hypothesized by Sipiczki and Kajdacsi (2009) based on the presence of Group I introns (Fig. 1; see discussion below). Since segregation of the families Volvocisporiaceae and Quambalariaceae appears unwarranted with our data, we take a conservative view in considering the order at present to contain a single family, Microstromataceae,

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until further data indicate a need for additional taxonomic partitioning. Group I introns Group I introns are conserved non-coding sequences that can perform self-splicing. They behave like transposable elements, and are likely to insert in rDNA genes within eukaryotic genomes (De Wachter et al. 1992). In Microstromatales, the group I intron is only known from the SSU region of Jaminaea angkorensis, which has been used to support a hypothesis of the early diverging placement for the genus (Sipiczki and Kajdacsi 2009). However, Jaminaea was described as a monotypic genus at the time of these studies. Here, we include more Jaminaea taxa and make a multiple sequence alignment of SSU sequences (Fig. S3). The results show that J. angkorensis is the only species of Jaminaea that possesses the SSU group I intron (Sicipzki and Kajdacsi 2009). It has to be noted that an SSU group I intron is also present in a few other species in Entylomatales and Exobasidiales (Exobasidiomycetes), as well as several species in Ustilaginomycetes (Sipiczki and Kajdacsi 2009; Fig. S3). A previous investigation in Agaricomycetes revealed that group I introns could be vertically or horizontally transmitted, depending on each lineage (Hibbett 1996). Broader taxon sampling is need to better resolve the origins of the group I intron insertion in Exobasidiomycete SSU sequences.

MycoBank: MB819463 Synonym: Jaminaea angkorensis Sipiczki & Kajdacsi, Int. J. Syst. Evol. Microbiol. 59(4):919 (2009) (Invalid by Art. 8.4 and 40.7, ICN) The description is that of Jaminaea angkorensis Sipiczki & Kajdacsi (2009). Holotype: CBS10918, permanently preserved in a metabolically inactive state at Centraalbureau voor Schimmelcultures Fungal Biodiversity. Ex-type culture: C5b ≡ CBS 10918 ≡ CCY 88-1-1. Jaminaea lanaiensis Mahdi, Statzell, Fell, M.V. Br. & Donachie ex T. Kij. & Aime, sp. nov. MycoBank: MB819464 Synonym: Sympodiomycopsis lanaiensis Mahdi, Statzell, Fell, M.V. Br. & Donachie, FEMS Yeast Res. 8(8):1359 (2008) (Invalid by Art. 8.4 and 40.7, ICN) ≡ Jaminaea lanaiensis (Mahdi, Statzell, Fell, M.V. Br. & Donachie) Liou, Y.H. Wei & F.L. Lee, Int. J. Syst. Evol. Microbiol. 61:471 (2011) (Invalid by Art. 35.1, ICN) The description is that of Sympodiomycopsis lanaiensis Mahdi et al. (2008). Holotype: CBS10858, permanently preserved in a metabolically inactive state at Centraalbureau voor Schimmelcultures Fungal Biodiversity. Ex-type culture: LM418 ≡ ATCC MYA4092 ≡ DSMZ 18755 ≡ NRRL Y-48466 ≡ CBS 10858.

Jaminaea angkorensis Sipiczki & Kajdacsi ex T. Kij. & Aime, sp. nov.

Jaminaea rosea T. Kij. & Aime, sp. nov. (Figs. 2b, c and 3) MycoBank: MB812676 Etymology: referring to the pink color of colonies on PDA. Type: USA, Florida, Coral Gables, a private garden, 25°43′ 34″N, 80°15′47″W, a Plumeria sp. leaf phylloplane infected with Coleosporium plumeriae, leg. M.C. Aime, 21 July 2013 (PUL F2907, holotype as a dried culture permanently preserved in a metabolically inactive state; MCA5214 ≡ NRRL 66311 ≡ CBS 14051, ex-type culture; GenBank accessions— SSU: KR912076, ITS: KR912071 and LSU: KR912073). Morphology: After incubation in YMB at 25 °C for 7 days, cells appear singly or in pairs with polar budding, subglobose or oval-shaped and ranging between 1.6–3.3 × 3.5–6.5 μm in size (Fig. 2b). Short pseudohyphae are present in the cultures (Fig. 2c). True hyphae are absent. Colonies on PDA after

Fig. 2 Microscopic morphologies of Pseudomicrostroma glucosiphilum and Jaminaea rosea. Each fungus was cultured in YMB and incubated for 7 days. Cell morphology was observed under ×100 magnification of a

differential interference contrast microscope. a Budding yeast cells of P. glucosiphilum. b Budding yeast cells and c Pseudohyphae structure of J. rosea. Bars 10 μm

Taxonomy Jaminaea Sipiczki & Kajdacsi ex T. Kij. & Aime, gen. nov. MycoBank: MB819462 Synonym: Jaminaea Sipiczki & Kajdacsi, Int. J. Syst. Evol. Microbiol. 59(4):919 (2009) (Invalid by Art. 40.1 and 40.3, ICN) The description is that of Jaminaea Sipiczki & Kajdacsi (2009). Type species: Jaminaea angkorensis Sipiczki & Kajdacsi ex T. Kij. & Aime

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members of Microstromatales are found within the temperate zone (Cooke 1962; Begerow et al. 2014), most records we could find for Jaminaea species seem to be confined to tropical and subtropical regions (Mahdi et al. 2008; Sipiczki and Kajdacsi 2009; Albu 2012).

Fig. 3 Colony morphologies of Pseudomicrostroma glucosiphilum (MCA4718) and Jaminaea rosea (MCA5214). Each isolate was cultured in four different types of media: PDA, CMA, YMA and YPGA. The cultures were incubated at room temperature for 14 days. Bars 2.5 mm

incubation at 25 °C for 14 days are cream-pink (oac 697), viscous, dull and smooth with a ciliate margin and flat elevation. Colonies on CMA after incubation at 25 °C for 14 days are pink (oac 578), mucoid, glistening to translucent with a filliform margin and flat elevation. Colonies on YMA after incubation at 25 °C for 14 days are cream-pink (oac 550), viscous, dull, wrinkled at the center and velvety at the border with a ciliate margin and convex to raised elevation. Colonies on YPGA after incubation at 25 °C for 14 days are creamwhite (oac 816), butyrous, smooth and glistening with an irregular margin and flat elevation (Fig. 3). Physiological characteristics: The following carbon compounds are assimilated: glucose, maltose, sucrose, mentiobiose (weak), maltotriose, palatinose, stachyose, turanose, cellobiose, gentobiose, lactose, L-sorbose, Dmelezitose, D-melibiose, D-raffinose, L-arabinose, D-ribose, D-xylose, α-methyl-D-glucoside, maltitol, galactitol (weak), D-mannitol, D-sorbitol, xylitol (weak), ierythritol, glycerol, methanol, ethanol, inositol (weak), 1,2-propanediol, succinic acid, L-glutamic acid, L-malic acid (weak), bromosuccinic acid, γ-aminnobutyric acid (weak), α-ketoglutarate (weak), D-gluconic acid (weak), D-glucuronate, DL-lactate, citrate and soluble starch. The following compounds are not assimilated: D-galactose, Darabinose, L-rhamnose, D-psicose, trehalose, salicin, Darabitol, ribitol and D-galacturonate. KNO3 (delayed) and NaNO2 (delayed) are assimilated as nitrogen sources. The fungus can grow at up to 37 °C, but not 39 °C. Fermentation is absent. Growth is observed in the presence of 60% glucose or 16% NaCl (weak). Gelatin liquefaction is positive. Starch-like substance is not produced. Notes: Jaminaea rosea has several physiological characteristics that readily distinguish it from the other three Jaminaea species. For instance, the ability to assimilate i-erythritol and lactose, as well as the ability to grow at 37 °C, distinguishes J. rosea from J. angkorensis and J. lanaiensis, while the inability to assimilate trehalose and ribitol distinguishes J. rosea from J. phylloscopi (Table 2). Interestingly, although most of

Microstroma bacarum (Buhagiar) T. Kij. & Aime, comb. nov. MycoBank: MB819465 Basionym: Torulopsis bacarum Buhagiar, J. gen. Microbiol. 86(1):2 (1975) ≡ Rhodotorula bacarum (Buhagiar) Rodr. Mir. & Weijman, Antonie van Leeuwenhoek 54(6):549 (1988) Parajaminaea T. Kij. & Aime, gen. nov. MycoBank: MB812677 Etymology: similar to Jaminaea Fungi belonging phylogenetically to Microstromatales (Exobasidiomycetes, Ustilaginomycotina, Basidiomycota). Anamorphs appear as budding yeasts which are ovoid to ellipsoidal. Pseudomycelia and hyphae have not been observed in culture, which is in contrast to members of Jaminaea. Teleomorphs are so far known as foliar phytopathogens on the woody plant genus Albizia. Type species: Parajaminaea albizii (Syd. & P. Syd.) T. Kij. & Aime. Parajaminaea albizii (Syd. & P. Syd.) T. Kij. & Aime, comb. nov. MycoBank: MB817111 Basionym: Microstroma albiziae Syd. & P. Syd., Annales Mycologici 12(3):54 (1914) Parajaminaea phylloscopi Francesca, Guerreiro, Carvalho, Sampaio & Moschetti ex T. Kij. & Aime, sp. nov. MycoBank: MB817617 Synonym: Jaminaea phylloscopi Francesca, Guerreiro, Carvalho, Sampaio & Moschetti, Int. J. Syst. Evol. Microbiol 66(2): 824–829 (2016) (Invalid by Art. 8.4, 35.1 and 40.7, ICN) The description is that of Jaminaea phylloscopi Francesca et al. (2016). Holotype: CBS14087, permanently preserved in a metabolically inactive state at Centraalbureau voor Schimmelcultures Fungal Biodiversity. Ex-type culture: 551B6 ≡ CBS 14087 ≡ PYCC 6783. Pseudomicrostroma T. Kij. & Aime, gen. nov. MycoBank: MB812674 Etymology: Bfalse^ Microstroma. Fungi belonging phylogenetically to Microstromatales (Exobasidiomycetes, Ustilaginomycotina, Basidiomycota). Anamorphs, where known, appear as budding yeasts which

Mycol Progress

are globose to ellipsoidal. Pseudomycelia and hyphae have not been observed in culture. All known species are unable to assimilate melibiose. Teleomorphs, where known, are foliar phytopathogens of woody plants in the genera Carya and Juglans that are morphologically similar to but phylogenetically distinct from Microstroma species. Type species: Pseudomicrostroma juglandis (Bérenger) T. Kij. & Aime. Pseudomicrostroma glucosiphilum T. Kij. & Aime, sp. nov. (Figs. 2a and 3) MycoBank: MB812675 Etymology: referring to its ability to grow on high glucose media. Type: Germany, Trier-Saarburg, Zemmer, 49°53′50″N, 6°42′ 1″E, isolated from PDA plates amended with 50% glucose exposed to air in a private house, leg. M.C. Aime, 2 April 2012 (PUL F2906, holotype as a dried culture permanently preserved in a metabolically inactive state; MCA4718 ≡ NRRL 66310 ≡ CBS 14053, ex-type culture; GenBank accessions—SSU: KR912075, ITS: KR912070 and LSU: KR912072). Morphology: After incubation in YMB at 25 °C for 7 days, cells appear singly or in pairs with polar budding, subglobose or ovoid-shaped and sized between 2.0–4.7 × 4.2–9.2 μm (Fig. 2a). Neither hyphae nor pseudohyphae are present in the cultures. Colonies on PDA after incubation at 25 °C for 14 days are cream (oac 801), butyrous, smooth and glistening with an entire margin and convex elevation. Colonies on CMA after incubation at 25 °C for 14 days are light pink (oac 578), mucoid, smooth and glabrous with a crenulate margin and convex elevation. Colonies on YMA after incubation at 25 °C for 14 days are cream (oac 814), butyrous, glistening, smooth and radially-ridged with an entire margin and convex elevation. Colonies on YPGA after incubation at 25 °C for 14 days are cream (oac 766), butyrous, rugulose and glistening with an entire margin and convex elevation (Fig. 3). Physiological characteristics: The following carbon compounds are assimilated: glucose, maltose, sucrose, trehalose, turanose, maltotriose, palatinose, stachyose, cellobiose (delayed), gentiobiose, D-raffinose, D-melezitose, D-arabinose, L-arabinose, D-ribose, D-xylose (delayed), α-methyl-Dglucoside (weak), maltitol, mannitol, D-sorbitol, xylitol (delayed), i-erythritol, glycerol, ethanol, inositol, 1,2-propanediol (weak), citrate, fumaric acid, L-malic acid, bromosuccinic acid, tween80 (delayed), dextrin and soluble starch (weak). The following carbon compounds are not assimilated: lactose, Dmelibiose, D-galactose, D-psicose, L-sorbose, D-glucuronate, D-galacturonate, DL-lactate, arbutin, salicin, ribitol, Darabitol and galactitol. KNO3, NaNO2 and cadaverine are assimilated as nitrogen sources. The fungus can grow at up to 30 °C, but not 35 °C. Growth is observed in the presence of 50% glucose or 10% NaCl (weak). Fermentation is absent.

Gelatin liquefaction is negative. Starch-like substance is not produced. Notes: Pseudomicrostroma glucosiphilum is distinguished from its sister species P. phylloplanum by the ability to grow under hyperosmotic conditions (50% glucose agar) and at elevated temperatures of up to 30 °C (Table 1). Pseudomicrostroma juglandis (Bérenger) T. Kij. & Aime, comb. nov. MycoBank: MB817110 Basionym: Fusidium juglandis Bérenger, Plantae rariores Carinthiacae: 7, fig. 1 (1847) ≡ Microstroma juglandis (Bérenger) Sacc., Syll. Fung. (Abellini) 4:9 (1886) = Torula juglandina Opiz, Seznam Rostlin Kveteny Ceské: 147 (1852) = Gymnosporium leucosporum Mont., Syll. Crypt. (Paris): 309 (1856) = Fusisporium pallidum Niessl, Verh. Zool.-bot. Ges. Wien 8(Abhandl): 329 (1858) ≡ Microstroma pallidum (Niessl) Niessl, Öst. bot. Z. 11:252 (1861) = Ascomyces juglandis Berk., Outl. Brit. Fung. (London): 376 (1860) = Microstroma juglandis var. robustum B.B. Higgins, Phytopathology 7:42 (1917) Pseudomicrostroma phylloplanum (R.G. Shivas & Rodr. Mir.) T. Kij. & Aime, comb. nov. MycoBank: MB812678 Basionym: Cryptococcus phylloplanus R.G. Shivas & Rodr. Mir., Antonie van Leewenhoek 49(2):153 (1983) ≡ Microstroma phylloplanum (R.G. Shivas & Rodr. Mir.) Q.M. Wang, F.Y. Bai, Begerow & Boekhout, Studies in Mycology 81:80 (2015) ≡ Rhodotorula phylloplana (R.G. Shivas & Rodr. Mir.) Rodr. Mir. & Weijman, Antonie van Leewenhoek. 54(6):549 (1988) = Cryptococcus hinnuleus R.G. Shivas & Rodr. Mir., Antonie van Leewenhoek. 49(2):155 (1983) Sympodiomycopsis kandeliae G.Y. Liou, Y.H. Wei & F.L. Lee ex T. Kij. & Aime, sp. nov. MycoBank: MB819466 Synonym: Sympodiomycopsis kandeliae G.Y. Liou, Y.H. Wei & F.L. Lee, Int. J. Syst. Evol. Microbiol. 61:472 (2011) (Invalid by Art. 8.4 and 40.7, ICN) The description is that of Sympodiomycopsis kandeliae G.Y. Liou, Y.H. Wei & F.L. Lee (2011). Holotype: CBS11676, permanently preserved in a metabolically inactive state at Centraalbureau voor Schimmelcultures Fungal Biodiversity. Ex-type culture: FIRDI 007 ≡ BCRC 23165 ≡ CBS 11676.

Mycol Progress

Sympodiomycopsis yantaiensis F.L. Hui, Liang Chen, Lin Zhang & Z.H. Li ex T. Kij. & Aime, sp. nov. MycoBank: MB819467 Synonym: Sympodiomycopsis yantaiensis F.L. Hui, Liang Chen, Lin Zhang & Z.H. Li, Int. J. Syst. Evol. Microbiol. 63:3503 (2013) (Invalid by Art. 8.4 and 40.7, ICN) The description is that of Sympodiomycopsis yantaiensis F.L. Hui, Liang Chen, Lin Zhang & Z.H. Li (2013). Holotype: CBS12813, permanently preserved in a metabolically inactive state at Centraalbureau voor Schimmelcultures Fungal Biodiversity. Ex-type culture: NYNU 121010 ≡ CICC 32998 ≡ CBS 12813.

Acknowledgements We thank the Plant and Pest Diagnostic Laboratory (PPDL) for microscopy and microplate facilities. Travel to Germany and Florida were facilitated by a travel grant from the International Mycological Congress and funding from CAPES to MCA, and we are grateful to M.R.A. DiBiasi and M. Dal Pan for providing lodging and gardens with interesting fungi. TK’s study at Purdue University is funded by an Anadamahidol scholarship from Thailand. We acknowledge Dr. Mehrdad Abbasi, all reviewers and editors for critical suggestions on systematic and nomenclatural issues. Finally, we would like to thank for all members of the Aime laboratory for help in culture maintenance and molecular laboratory support, as well as constructive comments on the previous versions of the manuscript.

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