- Open Access
Overview of Phacidiales, including Aotearoamyces gen. nov. on Nothofagus
© International Mycological Association 2018
- Received: 20 April 2018
- Accepted: 25 October 2018
- Published: 1 December 2018
The new genus Aotearoamyces is proposed to accommodate a single species that was repeatedly collected on fallen wood in Nothofagaceae forests of New Zealand and was previously misidentified as a Claussenomyces species. This monotypic genus belongs to Tympanidaceae, a recently erected family in Phacidiales. Aotearoamyces is differentiated from other Tympanidaceae by phragmospores that do not form conidia either in or outside the asci, an exciple of textura intricata with hyphae widely spaced and strongly gelatinized (plectenchyma), and apically flexuous. partly helicoid paraphyses. The asexual morph was studied in pure culture. Phylogenetic analyses of combined SSU, ITS and LSU sequences strongly support a sister relationship between the sexually typified Aotearoamyces and the asexually typified “Collophorina” paarla characterized morphologically by forming endoconidia, a feature not found in the genetically distinct type species of Collophorina. Based on our molecular results, we place the genus Epithamnolia in the Mniaecia lineage within Phacidiales.
- new taxa
The taxonomy and classification of the Leotiomycetes is unsettled with a high proportion of taxa not yet treated using molecular methods (Baral 2016, LoBuglio & Pfister 2010). Consequently, the delimitation of genera, families and orders often changes and the systematic position of taxa is subject to modification, depending on individual researcher’s opinions, the impact of the addition of sequences from previously unsampled taxa, opinions about the acceptance of paraphyletic groups, etc. One of the orders recognized in the class is Phacidiales. A study of the genera considered to belong in this order provides a good example of the chaotic situation within the class.
How the number of genera and their systematic placement, which reflects changes in morphological concepts, has changed over time is illustrated in Fig. 1. Höhnel (1917) described the Phacidiales as follows: “Discomycetes with superficial or immersed fruitbodies, never erumpent fruitbodies, with or without stroma, excipulum entire or only at the margin, thin and brown or thick and carbonaceous. At maturity it opens very irregularly by a longitudinal split or by several lobes. Rarely the covering layer over the hymenium forms a detaching lid”. After Höhnel’s circumscription, the sexual morph was usually described as a reduced carbonaceous ascoma: black, discoid to hysteriform, frequently immersed in the tissue of the host and with a reduced exciple. The hymenium was described as exposed by a rupture of the upper stromatal layer by one or more slits. The asci were reported as 4–8-spored, thickened apically, with or without an amyloid apical ring. The ascospores were referred to as variable in shape, simple or phragmoseptate, hyaline or rarely brownish, and with or without mucilaginous sheaths or appendages (e.g. Ainsworth & Bisby 1943, 1950, Ainsworth 1961, Korf 1973, Dennis 1978). The concept of the order that developed after Höhnel generally included three families (Fig. 1): Cryptomycetaceae, Hypodermataceae or Rhytismataceae, and Phacidiaceae (e.g. Ainsworth et al. 1971, Korf 1973, Dennis 1978, DiCosmo et al. 1984).
Between 1983 and 1995 the order Phacidiales fell out of use, and the family Phacidiaceae was applied in a more restricted sense, including Phacidium and two or three other genera. Some authors even considered Phacidiales as a synonym of Rhytismatales (Hawksworth et al. 1983). In other cases, Phacidiaceae, Cryptomycetaceae and Rhytismataceae were included in Rhytismatales (Hawksworth et al. 1983, 1995). Rhytismataceae and Hypodermataceae included many genera previously placed in Phacidiales (Hawksworth et al. 1983, 1995). In 1995 the family Phacidiaceae contained three genera and was placed by Korf & Lizoň (2000) in Leotiales, an invalid name later validated by Korf & Lizoň (2000), and there in Helotiales. In 2001 the Phacidiaceae still contained only three genera (Ascocoma, Lophophacidium, and Phacidium) and was transferred to Helotiales, where it was treated during 2001–2010 (Kirk et al. 2001, 2008, Eriksson 2005, 2006, Lumbsch & Hundorf 2007, 2010).
Crous et al. (2014) recognized the Phacidiales as a monophyletic order distinct from Helotiales and included six genera (Fig. 1). Using molecular evidence, these authors expanded the morphological concept of the order by including genera with exposed, cup-shaped apothecia typical of helotiaceous fungi (e.g. Bulgaria) as well as genera with immersed ascomata that open by splits across covering stromatic layers, as was characteristic of the concepts of Phacidiales of earlier authors. Although DiCosmo et al. (1984) reported anamorphs for some members in Phacidiales, it was Crous et al. (2014) who provided a unified view of the asexual morphs within the order. Previously, the information about asexual morphs was sparse and only a relationship with coelomycetes had been reported (DiCosmo et al. 1984).
In the most recent classification of Leotiomycetes compiled by Baral (2016), the ecology of the order remained the same (saprobic, parasitic), but the morphological concept was expanded and delineated more precisely, including information about the phase during which the apothecia open (prohymenial to mesohymenial), and added features of the living cells, such as the lack of vacuolar bodies in paraphyses, asci with either amyloid or inamyloid apical rings (exceptionally the entire wall is amyloid) and ascospores with variable lipid content. Here the order Phacidiales has three families containing about 27 genera, approximately half the number of genera compared to Höhnel’s concept a century ago (Fig. 1). Two to three genera were added to Phacidiaceae in addition to those considered by Crous et al. (2014): Darkera, Starbaeckia, and questionably Gremmenia. Also, the priority of Phacidiopycnis over the sexually typified Potebniamyces was indicated. Two new families were included in the order: Tympanidaceae and Helicogoniaceae. In addition to these three families in Phacidiales, Baral included the ‘Mniaecia lineage’ with one or two genera (Mniaecia, and ?Trizodia), and one genus as incertae sedis (Coma with the sexually typified synonym Ascocoma). Subsequently, Suija et al. (2017) placed the monotypic genus Epithamnolia as incertae sedis in Phacidiales, due to its phylogenetic and morphological affinities with the asexual morph of Epiglia (a synonym of Mniaecia fide Van Vooren 2005), thus widening the ecological concept of the order to include lichenicolous fungi.
Taking into consideration the repeated changes within Phacidiales, the aim of this research was to enhance and synthesize knowledge of the order. Important results include the erection of a new genus known only from the Southern Hemisphere for a species previously misclassified in Claussenomyces, and the observation that the asexual “Collophorina” paarla is related to it.
Specimens of the newly described species were collected between 1989 and 2010 in native forests of New Zealand during non-targeted, general collecting expeditions for fungi. All specimens cited are deposited in the PDD fungarium (Manaaki Whenua Landcare Research, Auckland) and living cultures grown from ascospores from the fresh specimens are stored in the ICMP culture collection (Manaaki Whenua Landcare Research, Auckland, www.landcareresearch.co.nz/resources/collections/icmp).
Sections for anatomical examination of ascomata were free-hand sectioned under a Motic stereomicroscope SMZ140 and examined with a Motic B1 light microscope. Microphotographs were taken with an USB Moticam 2500 camera and processed with the software Motic images Plus 2.0. Measurements are given as follows: (smallest single measurement) smallest mean-largest mean (largest single measurement). The small and large means are based on ≥10 measurements of individual specimens. No living specimens of the sexual morph were available, and therefore potassium hydroxide at 5 % (KOH) was used to rehydrate herbarium specimens prior to morphological study. Conidia and conidiogenous cells were measured from dried Oatmeal Agar cultures rehydrated in 5 % KOH. The descriptions and abbreviations follow Baral (1992): † = dead state, × = living state; LBs = lipid bodies. Colour coding refers Anonymous (1976).
Specimens used in this studywith family information and GenBankaccession numbers. Sequences of the newspecies are indicated in bold.
Sarcoleotia globosa 1
Sarcoleotia globosa 2
Epithamnolia on Candelaria (HA90)
incertae sedis s. Suija et al. (2017)
Epithamnolia on Lecanora (HA92)
incertae sedis s. Suija et al. (2017)
Trizodia acrobia 1
Trizodia acrobia 2
Geltingia associata 1
Geltingia associata 2
Eleutheromyces subulatus 1
Eleutheromyces subulatus 2
Eleutheromyces subulatus 3
Collophorina africana 1
Collophorina africana 2
Collophorina paarla 1
Collophorina paarla 2
Holwaya mucida 1
Holwaya mucida 2
Aotearoamyces nothofagi ICMP 21969
Aotearoamyces nothofagi ICMP 21968
Aotearoamyces nothofagi PDD 106298
An analysis using three different rDNAregions (SSU, ITS, LSU) for the representative members of Phacidiales was performed. This includes taxa from three families: Phacidiaceae (5 seq.), Helicogoniaceae (5 seq.) and Tympanidaceae (12 seq.). Also, five sequences of the Mniaecia lineage were included, and two representing the genus Epithamnolia, which was recently placed in Phacidiales as incertae sedis (Suija et al. 2017). Thirty-one taxa were used for the molecular analysis (Table 1). The sequences were aligned using the L-INS-i algorithm for the ITS region, and G-INS-i algorithm for SSU & LSU (Katoth & Toh 2008) with MAFFT v7.017 (Katoh et al. 2002). The program Gblocks v. 0.91b was used to identify and eliminate ambiguously aligned regions (Castresana 2000), using the following relaxed settings (Talavera & Castresana 2007): minimum number of sequences for a conserved or flanking position= 16; maximum number of contiguous non-conserved position= 10; minimum length of a block= 5; and gaps in an alignment column allowed in up to half the number of included sequences. The analyses were performed using the optimal model of nucleotide substitution identified with JModeltest (Posada 2008; http://darwing.uvigo.es), based on the Akaike information criterion (Akaike 1974). Maximum likelihood (ML) and Bayesian Inference (BI) analyses were performed using Geneious v.6.1.7. Bayesian inference analyses followed Quijada et al. (2014), only varying in the number of starting trees (10 million generations) and the tree sampling (every 1000th generation) for BI analysis. Branch support in ML was inferred from 1000 rounds of bootstrap. We only considered supported clades for ML those with bootstraps values ≥75 % and with PP≥0.95 (strongly supported) for BI. Phylogenetics trees were drawn with Geneious and artwork was prepared in Adobe Illustrator CS5.
Aotearoamyces P.R. Johnst., J.A. Cooper & Quijada, gen. nov.
Etymology: The generic name refers to the indigenous name of New Zealand (Aotearoa) and the Greek name for fungi (myces).
Diagnosis: The sexual morph of Aotearoamyces resembles Holwaya mucida, but the apothecia are turbinate with the disc plane or slightly convex. Ascus and ascospore shape are similar to species of Claussenomyces, but without production of conidia or ascoconidia directly from the ascospores. Durandiella and Tympanis have similar exciples, but Aotearoamyces differs in hyphae that are strongly spaced and gelatinized. It differs from all the others members in Tympanidaceae by the curved or helicoid paraphyses. The asexual morph lacks endoconidia in the vegetative hyphae; conidiophores occur in well-developed synnemata; and conidia are small, 0-septate, hyaline and curved, formed by phialidic conidiogenesis.
Type species: Aotearoamyces nothofagi P.R. Johnst. et al. 2018
Classification: Tympanidaceae, Phacidiales, Leotiomycetes, Pezizomycotina, Ascomycota, Fungi.
Description: Ascomata apothecia, black, erumpent, short to medium long stipitate (to 1 mm tall), pulvinate-discoid to turbinate, solitary, or more commonly clustered in groups and arising from a common gelatinous stromatic base. Asci 8-spored, slightly thick-walled towards apex, inamyloid, arising from croziers. Ascospores cylindric-fusoid to fusoid-clavate and phragmoseptate, rarely with a longiseptum. Paraphyses apically flexuous to helicoid or curving downwards (hooked) and embedded in an olive-brown gelatinous matrix. Exciple of textura intricata, innermost layer of the exciple composed of a loose network of narrow hyphae, widely spaced and embedded in an abundant light brown gelatinous matrix (plectenchyma); outer ectal exciple with pustules composed of closely septate, prismatic to angular cells, dark brownish, cells more densely packed than in the inner ectal exciple, cells covered by a dark brown pigmented exudate. Asexual morph in culture with short-cylindric, curved, 0-septate, hyaline conidia formed at a single, apical conidiogenous locus on flask-shaped, phialidic conidiogenous cells. Conidiogenous cells solitary or with several cells held on a single, short, cylindric basal cell, on hyphae grouped into ropey, synemmatous structures.
Aotearoamyces nothofagi P.R. Johnst., J.A. Cooper & Quijada, sp. nov.
Etymology: The specific epithet refers to the generic name of the host plant in the holotype (Nothofagus).
Diagnosis: Apothecia black, to 1 mm diam and height, erumpent, short to medium long stipitate, pulvinate-discoid to turbinate, arising from a common gelatinous stromatic base. Asci †83–124 × 10.5–14.5 µm, 8-spored, inamyloid, arising from croziers. Ascospores †17.5–31 × 3–5 µm, cylindric-fusoid-clavate, 7–16 phragmoseptate rarely with a longiseptum. Paraphyses apically up to 11.5–2(2.5) µm wide, flexuous to helicoid or curving downwards (hooked) and embedded in an olive-brownish gelatinous matrix. Conidia observed in culture, produced from phialides. Conidiogenous cells held on well-developed conidiophores arranged in small synnematous structures, forming consistently curved vermiform conidia. Conidiomata not observed in cultures.
Type: New Zealand: South Island: Craigiebrun, on Nothofagus solandri, 7 May 2010, N. Siegel (PDD 95741 — holotype).
Other specimens examined: New Zealand: South Island: Abel Tasman National Park, on unidentified wood, 14 May 2004, P.R. Johnston D1844 (PDD 80575, ICMP 21037); Arthur’s Pass National Park, on unidentified wood, 5 May 1989, P.R. Johnston D368, G.L. Barron, P.K. Buchanan & M. Rajchenberg (PDD 55517, ICMP 21038); Otago Lakes, Routeburn Track carpark, on unidentified fallen wood in Nothofagaceae forest, 7 May 2016, S. McMullan-Fisher (PDD 110269).
Throughout its history, the number of species, genera and families in the order Phacidiales has changed considerably (Fig. 1). The order as circumscribed by Bessey (1907), who included six genera and three families, was differently conceived by Höhnel (1917), who expanded the order to include 52 genera in six families. In the 1970s (e.g. Korf 1973, Dennis 1978) the rhytismataceous fungi were often included in Phacidiales, although today they are placed in the separate order Rhytismatales. The most current classification of Phacidiales includes about 29 genera, most of them distributed across three families and one informal taxonomic lineage (Crous et al. 2014, Baral 2016, Suija et al. 2017). These changing concepts reflect the changes in emphasis placed on macro- and micromorphological features, as well as the impact of molecular phylogenetics. Molecular studies have allowed genera known only from an asexual morph, such as Collophorina, to be placed in Phacidiales (Baral loc. cit.). Our phylogenetic analyses allowed placement of Epithamnolia, a conidial fungus previously reported as incertae sedis in Phacidiales (Suija et al. 2017), in the Mniaecia clade for the first time.
Five species, known only from asexual morphs that were isolated from woody necroses in peach and nectarine, were included when Damm et al. (2010) erected the genus Collophora with C. africana, C. capensis, C. paarla, C. pallida, and C. rubra, the type species). Since that name was illegitimate as a later homonym of Collophora Mart. 1830, Apocynaceae, the species were recombined into the new genus Collophorina, and the number of species reduced from seven to five due to synonymy of C. capensis with C. africana, and C. pallida with C. paarla (Wijayawardene et al. 2017). Damm et al. (2010) placed the genus in Leotiomycetes as incertae sedis. In the same work, the authors remarked “although these species form two clades in the LSU phylogeny, they are placed in one genus, because of their similar morphological features and the lack of morphological characters distinguishing the two clades”. In our analyses, the genus is also paraphyletic in agreement with Damm et al. (2010) (Fig. 2): Collophorina paarla belongs in one supported clade (Fig. 2, Clade J), and C. africana and the type species C. rubra in a different strongly supported clade.
In the discussion about C. pallida, Damm et al. (2010) said that “C. paarla and C. pallida are the only Collophora species for which endoconidia have been observed”. This morphological feature could be used to support the splitting of Collophorina into at least two genera. Aotearoamyces is most closely related to the clade containing the Collophorina species with endoconidia, but we did not see any endoconidia form in our culture studies. Compared to Damm et al.’s illustrations and descriptions, the conidiogenus cells of Aotearomyces nothofagi are held on well-developed synnematous conidiophores bearing conidia that are consistently curved.
L.Q. thanks the “Fundación Ramón Areces” for support. This study is part of the project “DNA barcoding for plant-pathogens diagnostic and monitoring: Forest diseases and turbo-taxonomy in Tympanidaceae as a case of study”, and also the fellowship programme Becas Fundación Ramón Areces para Estudios Postdoctorales, XXIX Convocatoria para Ampliación de Estudios en el Extranjero en Ciencia de la Vida y de la Materia. P.R.J. and J.A.C. were supported with funding from the Science and Innovation Group of the New Zealand Ministry of Business, Innovation and Employment through the Manaaki Whenua Landcare Research Systematics Portfolio. Also, we would like to thank Hans-Otto Baral for revising the manuscript.
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
- Ainsworth GC, Bisby G.R. (1943) A Dictionary of the Fungi. Kew: Imperial Mycological Institute.Google Scholar
- Ainsworth GC, Bisby GR (1950) A Dictionary of the Fungi. 3rd edn. Kew: Commonwealth Mycological Institute.Google Scholar
- Ainsworth GC (1961) Ainsworth & Bisby’s Dictionary of the Fungi. 5th edn. Kew: Commonwealth Mycological Institute.Google Scholar
- Ainsworth GC, James PW, Hawksworth DL (1971) Ainsworth & Bisby’s Dictionary of the Fungi. 6th edn. Kew: Commonwealth Mycological Institute.Google Scholar
- Akaike H (1974) A new look at the statistical model identification. IEEE Transactions on Automatic Control 19: 716–723.View ArticleGoogle Scholar
- Anonymous (1976) ISCC-NBS Color-name Charts illustrated with centroid colors. Washington DC: Inter-Society Color Council, National Bureau of Standards.Google Scholar
- Baral HO (1992) Vital versus herbarium taxonomy: morphological differences between living and dead cells of ascomycetes, and their taxonomic implications. Mycotaxon 44: 333–390.Google Scholar
- Baral HO (2016) Inoperculate discomycetes. In: Syllabus of Plant families. A. Engler’s Syllabus der Pflanzenfamilien. Vol. 1(2). Ascomycota (Jaklitsch W, Baral HO, Lücking R, Lumbsch T, eds): 55–221. 13th edn. Stuttgart: Borntraeger Science Publishers.Google Scholar
- Bessey CE (1907) A synopsis of plant phyla. University of Nebraska Studies 7: 275–373.Google Scholar
- Bunyard BA, Nicholson MS, Royse DJ (1994) A systematic assessment of Morchella using RFLP analysis of the 28S ribosomal RNA gene. Mycologia 86: 762–772.View ArticleGoogle Scholar
- Castresana J (2000) Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. Molecular and Evolution 17: 540–552.View ArticleGoogle Scholar
- Crous PW, Quaedvlieg W, Hansen K, Hawksworth DL, Groenewald JZ (2014) Phacidium and Ceuthospora (Phacidiaceae) are congeneric: taxonomic and nomenclatural implications. IMA Fungus 5: 173–193.View ArticleGoogle Scholar
- Damm U, Fourie PH, Crous PW (2010) Coniochaeta (Lecythophora), Collophora gen. nov. and Phaeomoniella species associated with wood necroses of Prunus trees. Persoonia 24: 60–80.View ArticleGoogle Scholar
- Dennis RWG (1978) British Ascomycetes. 2nd edn. Vaduz: J. Cramer.Google Scholar
- DiCosmo F, Nag Raj TR, Kendrick WB (1984) A revision of the Phacidiaceae and related anamorphs. Mycotaxon 21: 1–234.Google Scholar
- Eriksson OE (2005) Outline of Ascomycota - 2005. Myconet 11: 1–113.Google Scholar
- Eriksson OE (2006) Outline of Ascomycota - 2006. Myconet 12: 1–82.Google Scholar
- Hawksworth DL, Kirk PM, Sutton BC, Pegler DN (1995) Ainsworth & Bisby’s Dictionary of the Fungi. 8th edn. Wallingford: CAB International.Google Scholar
- Hawksworth DL, Sutton BC, Ainsworth GC (1983) Ainsworth & Bisby’s Dictionary of the fungi (including the lichens). 7th edn. Kew: Commonwealth Mycological Institute.Google Scholar
- Höhnel F (1917) System der Phacidialesv. H. Berichte der Deutschen Botanischen Gesellschaft 34: 416–422.Google Scholar
- Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes — application to the identification of mycorrhizae and rusts. Molecular Ecology 2: 113–118.View ArticleGoogle Scholar
- Johnston PR, Park D (2013) The phylogenetic position of Lanzia berggrenii and its sister species. Mycosystema 32: 366–385.Google Scholar
- Katoh K, Misawa K, Kuma K, Miyata T (2002) MAFFT, a novel method for rapid multiple sequence alignment base don fast Fourier transform. Nucleic Acid Research 30: 3059–3066.View ArticleGoogle Scholar
- Katoh K, Toh H (2008) Improved accuracy of multiple ncRNA alignment by incorporating structural information into a MAFFT-based framework. BMC Bioinformatics 9: 212.View ArticleGoogle Scholar
- Kirk PM, Cannon PF, David JC, Stalpers JA (2001) Ainsworth & Bisby’s Dictionary of the Fungi. 9th edn. Wallingford: CAB International.Google Scholar
- Kirk PM, Cannon PF, Minter DW, Stalper JA (2008) Ainsworth & Bisby’s Dictionary of the Fungi. 10th edn. Wallingford: CAB International.View ArticleGoogle Scholar
- Korf RP (1973) Discomycetes and Tuberales. In: The Fungi: an advanced treatise. Vol. IVA. A Taxonomic Review with keys: Ascomycetes and Fungi Imperfecti (Ainsworth GC, Sparrow FK, Sussman AS, eds): 249–319. New York: Academic Press.Google Scholar
- Korf RP, Lizoň P (2000) Validation of Nannfeldt’s ordinal name Helotiales. Mycotaxon 75: 501–502.Google Scholar
- LoBuglio KF, Pfister DH (2010) Placement of Medeolaria farlowii in the Leotiomycetes, and comments on sampling within the class. Mycological Progress 9: 36–368.View ArticleGoogle Scholar
- Lumbsch HT, Huhndorf SM (2007) Outline of Ascomycota - 2007. Myconet 13:1–58.Google Scholar
- Lumbsch HT, Huhndorf SM (2010) Outline of Ascomycota - 2009. Myconet 14:1–40.Google Scholar
- Posada D (2008) jModelTest: phylogenetic model averaging. Molecular Biology and Evolution 25: 1253–1256.View ArticleGoogle Scholar
- Quijada L, Baral HO, Jaen-Molina R, Weiss M, Caujapé-Castells J, Beltrán-Tejera E (2014) Phylogenetic and morphological circumscription of the Orbilia aurantiorubra group. Phytotaxa 175: 1–18.View ArticleGoogle Scholar
- Quijada L (2015) Estudio de los órdenes Helotiales s.l. y Orbiliales (Ascomycota, Fungi) en la Isla de Tenerife. PhD thesis, Universidad de La Laguna.Google Scholar
- Suija A, van den Boom P, Zimmermann E, Zhurbenko MP, Diederich P (2017) Lichenicolous species of Hainesia belong to Phacidiales (Leotiomycetes) and are included in an extended concept of Epithamnolia. Mycologia 109: 882–899.View ArticleGoogle Scholar
- Talavera G, Castresana J (2007) Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Systematic Biology 56: 564–577.View ArticleGoogle Scholar
- Van Vooren N (2005) Mniaecia gloeocapsae (Boud.) van Vooren. Bulletin Mensuel de la Société Linnéenne de Lyon 74: 19.View ArticleGoogle Scholar
- Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238–4246.View ArticleGoogle Scholar
- White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols: a guide to methods and applications (Innis MA, Gelfand DH, Sninsky JJ & White TJ, eds): 315–322. San Diego: Academic Press.Google Scholar
- Wijayawardene NN, Hyde KD, Rajeshkumar KC, Hawksworth DL, Madrid H, et al. (2017) Notes for genera Ascomycota. Fungal Diversity 86: 1–594.View ArticleGoogle Scholar