Mycological Community Research Articles

Proposals for consideration at IMC12 to modify provisions related solely to fungi in Chapter F of the International Code of Nomenclature for algae, fungi, and plants

Seven proposals or sets of proposals to modify the provisions of Chapter F of the International Code of Nomenclature for algae, fungi, and plants (ICNafp) have been received. These proposals are formally presented together here. The topics addressed relate to: fungi whose morph-names have the same epithet; the listing of synonyms under entries for protected names in the Code Appendices; the processes of protection and rejection; the use of DNA sequences as nomenclatural types; the use of genomes as nomenclatural types; and the designation of fungi known only from DNA sequences. In addition, a suggestion is included to update the mention of the World Directory of Culture Collections in Article 40.7 Note 4. A Synopsis of the formal proposals will be provided in early July 2024, and the mycological community will be invited to provide a guiding vote on the proposals with a closing date of 2 August 2024. Final decisions on the proposals will be made following debate at the Fungal Nomenclature Session of IMC12 in August 2024.

What are the 100 most cited fungal genera?

The global diversity of fungi has been estimated between 2 to 11 million species, of which only about 155 000 have been named. Most fungi are invisible to the unaided eye, but they represent a major component of biodiversity on our planet, and play essential ecological roles, supporting life as we know it. Although approximately 20 000 fungal genera are presently recognised, the ecology of most remains undetermined. Despite all this diversity, the mycological community actively researches some fungal genera more commonly than others. This poses an interesting question: why have some fungal genera impacted mycology and related fields more than others? To address this issue, we conducted a bibliometric analysis to identify the top 100 most cited fungal genera. A thorough database search of the Web of Science, Google Scholar, and PubMed was performed to establish which genera are most cited. The most cited 10 genera are Saccharomyces, Candida, Aspergillus, Fusarium, Penicillium, Trichoderma, Botrytis, Pichia, Cryptococcus and Alternaria. Case studies are presented for the 100 most cited genera with general background, notes on their ecology and economic significance and important research advances. This paper provides a historic overview of scientific research of these genera and the prospect for further research. Citation: Bhunjun CS, Chen YJ, Phukhamsakda C, Boekhout T, Groenewald JZ, McKenzie EHC, Francisco EC, Frisvad JC, Groenewald M, Hurdeal VG, Luangsa-ard J, Perrone G, Visagie CM, Bai FY, Błaszkowski J, Braun U, de Souza FA, de Queiroz MB, Dutta AK, Gonkhom D, Goto BT, Guarnaccia V, Hagen F, Houbraken J, Lachance MA, Li JJ, Luo KY, Magurno F, Mongkolsamrit S, Robert V, Roy N, Tibpromma S, Wanasinghe DN, Wang DQ, Wei DP, Zhao CL, Aiphuk W, Ajayi-Oyetunde O, Arantes TD, Araujo JC, Begerow D, Bakhshi M, Barbosa RN, Behrens FH, Bensch K, Bezerra JDP, Bilański P, Bradley CA, Bubner B, Burgess TI, Buyck B, Čadež N, Cai L, Calaça FJS, Campbell LJ, Chaverri P, Chen YY, Chethana KWT, Coetzee B, Costa MM, Chen Q, Custódio FA, Dai YC, Damm U, de Azevedo Santiago ALCM, De Miccolis Angelini RM, Dijksterhuis J, Dissanayake AJ, Doilom M, Dong W, Alvarez-Duarte E, Fischer M, Gajanayake AJ, Gené J, Gomdola D, Gomes AAM, Hausner G, He MQ, Hou L, Iturrieta-González I, Jami F, Jankowiak R, Jayawardena RS, Kandemir H, Kiss L, Kobmoo N, Kowalski T, Landi L, Lin CG, Liu JK, Liu XB, Loizides M, Luangharn T, Maharachchikumbura SSN, Makhathini Mkhwanazi GJ, Manawasinghe IS, Marin-Felix Y, McTaggart AR, Moreau PA, Morozova OV, Mostert L, Osiewacz HD, Pem D, Phookamsak R, Pollastro S, Pordel A, Poyntner C, Phillips AJL, Phonemany M, Promputtha I, Rathnayaka AR, Rodrigues AM, Romanazzi G, Rothmann L, Salgado-Salazar C, Sandoval-Denis M, Saupe SJ, Scholler M, Scott P, Shivas RG, Silar P, Souza-Motta CM, Silva-Filho AGS, Spies CFJ, Stchigel AM, Sterflinger K, Summerbell RC, Svetasheva TY, Takamatsu S, Theelen B, Theodoro RC, Thines M, Thongklang N, Torres R, Turchetti B, van den Brule T, Wang XW, Wartchow F, Welti S, Wijesinghe SN, Wu F, Xu R, Yang ZL, Yilmaz N, Yurkov A, Zhao L, Zhao RL, Zhou N, Hyde KD, Crous PW (2024). What are the 100 most cited fungal genera? Studies in Mycology 108: 1-411. doi: 10.3114/sim.2024.108.01.

How, not if, is the question mycologists should be asking about DNA-based typification.

Fungal metabarcoding of substrates such as soil, wood, and water is uncovering an unprecedented number of fungal species that do not seem to produce tangible morphological structures and that defy our best attempts at cultivation, thus falling outside the scope of the International Code of Nomenclature for algae, fungi, and plants. The present study uses the new, ninth release of the species hypotheses of the UNITE database to show that species discovery through environmental sequencing vastly outpaces traditional, Sanger sequencing-based efforts in a strongly increasing trend over the last five years. Our findings challenge the present stance of some in the mycological community - that the current situation is satisfactory and that no change is needed to "the code" - and suggest that we should be discussing not whether to allow DNA-based descriptions (typifications) of species and by extension higher ranks of fungi, but what the precise requirements for such DNA-based typifications should be. We submit a tentative list of such criteria for further discussion. The present authors hope for a revitalized and deepened discussion on DNA-based typification, because to us it seems harmful and counter-productive to intentionally deny the overwhelming majority of extant fungi a formal standing under the International Code of Nomenclature for algae, fungi, and plants.

Fouling communities of microscopic fungi on various substrates of the Black Sea

Fungi are the most active biodeteriorators of natural and man-made materials. The article presents generalizations of the studies (2001–2019) of communities of microscopic fungi within biofilms on various substrates: shells of live Mytilus (Mytilus galloprovincialis, 670 specimens) and Ostreidae (Crassostrea gigas, 90 specimens), fragments of driftwood (over 7,000), stones (40), concrete of hydrotechnical constructions along the shoreline (80) and wood between concrete blocks in constructions on the shores (80). The studies were carried out in Odessa Oblast, the coastal zone of Sevastopol and open area of the Black Sea. There were identified 123 species of micromycetes, belonging to 65 genera, 33 families, 21 orders, 10 classes, 4 divisions, 2 kingdoms: Fungi and Chromista (fungi-like organisms). The Chromista kingdom was represented by 1 species – Ostracoblabe implexa, on shells of C. gigas. The number of species of micromycetes on various substrates varied 23 (wood between concrete blocks of hydrotechnical constructions) to 74 (shells of M. galloprovincialis at the depths of 3 and 6 m). On all the substrates, the following species were found; Alternaria alternata, Botryotrichum murorum. The communities were found to contain pathogenic fungi Aspergillus fumigatus (shells of mollusks, stones, concrete), A. terreus (concrete), Fusarium oxysporum, Pseudallescheria boydii (shells of mollusks). The best representation was seen for the Pleosporales order – from 12.9% (shells of M. galloprovincialis, 0.3 m depth) to 33.3% (shells of C. gigas) of the species composition. Toxin-producing species of Microascales in mycological communities accounted for 1.6% (driftwood) to 40.0% (concrete), and were also observed on shells of Bivalvia – 11.1–32.3%. Similarity of species composition of mycological communities according to Bray-Curtis coefficient varied 21.1% (driftwood and concrete, 10 shared species) to 72.7% (shells of M. galloprovincialis, the depths of 3 and 7 m and shells of C. gigas, 45 shared species). Using graphs of indices of mean taxonomic distinctness (AvTD, Δ+) and variation (Variation in Taxonomic Distinctness index, VarTD, Λ+), we determined deviations of taxonomic structure of the studied mycological communities from the level of mean expected values, calculated based on the list of species, taking into account their systematic positions. The lowest values of index Δ+ were determined for communities on shells of M. galloprovincialis, 0.3 m depth, driftwood, stones and concrete. These communities had uneven distribution of species according to higher taxonomic ranks and minimum number of the highest taxa: 4–6 classes, 1–2 divisions, Fungi kingdom. Disproportion in species composition with decrease in the number of the highest taxa occurred in extreme environmental conditions. Using index Λ+, we found that the most complex taxonomic structure of fungi communities has developed on concrete and shells of C. gigas. In mycological communities on those substrates, the number of species was low (25 and 46), but they belonged to 4–7 classes, 2–3 divisions, 1–2 kingdoms. To compare the structures of mycological communities that have developed in such substrates in biotopes sea, sea-land-air, land-air, we compiled a list of fungi based on the literature data, which, taking into account our data, comprised 445 species of 240 genera, 103 families, 51 orders, 15 classes, 5 divisions, 2 kingdoms. The analysis revealed that on substrates with similar chemical composition, in all the biotopes, the species of the same divisions dominated (genus and family may vary). Therefore, in the biotope land-air – Hypocreales, Pleosporales, Eurotiales (genera Acremonium, Fusarium, Alternaria, Aspergillus, Penicillium); sea – Pleosporales, Eurotiales, Microascales (Alternaria, Aspergillus, Penicillium, Corollospora); sea-land-air – Pleosporales, Microascales (Alternaria, Leptosphaeria, Aspergillus, Penicillium, Corollospora, Halosarpheia). Monitoring of species composition of myxomycetes is needed in farms that cultivate industrial objects, recreation sites, various buildings for prevention of mycotoxin intoxication and infestation by mycodermatoses and other diseases caused by opportunistic and pathogenic fungi.

Fouling communities of microscopic fungi on various substrates of the Black Sea

Fungi are the most active biodeteriorators of natural and man-made materials. The article presents generalizations of the studies (2001–2019) of communities of microscopic fungi within biofilms on various substrates: shells of live Mytilus (Mytilus galloprovincialis, 670 specimens) and Ostreidae (Crassostrea gigas, 90 specimens), fragments of driftwood (over 7,000), stones (40), concrete of hydrotechnical constructions along the shoreline (80) and wood between concrete blocks in constructions on the shores (80). The studies were carried out in Odessa Oblast, the coastal zone of Sevastopol and open area of the Black Sea. There were identified 123 species of micromycetes, belonging to 65 genera, 33 families, 21 orders, 10 classes, 4 divisions, 2 kingdoms: Fungi and Chromista (fungi-like organisms). The Chromista kingdom was represented by 1 species – Ostracoblabe implexa, on shells of C. gigas. The number of species of micromycetes on various substrates varied 23 (wood between concrete blocks of hydrotechnical constructions) to 74 (shells of M. galloprovincialis at the depths of 3 and 6 m). On all the substrates, the following species were found; Alternaria alternata, Botryotrichum murorum. The communities were found to contain pathogenic fungi Aspergillus fumigatus (shells of mollusks, stones, concrete), A. terreus (concrete), Fusarium oxysporum, Pseudallescheria boydii (shells of mollusks). The best representation was seen for the Pleosporales order – from 12.9% (shells of M. galloprovincialis, 0.3 m depth) to 33.3% (shells of C. gigas) of the species composition. Toxin-producing species of Microascales in mycological communities accounted for 1.6% (driftwood) to 40.0% (concrete), and were also observed on shells of Bivalvia – 11.1–32.3%. Similarity of species composition of mycological communities according to Bray-Curtis coefficient varied 21.1% (driftwood and concrete, 10 shared species) to 72.7% (shells of M. galloprovincialis, the depths of 3 and 7 m and shells of C. gigas, 45 shared species). Using graphs of indices of mean taxonomic distinctness (AvTD, Δ+) and variation (Variation in Taxonomic Distinctness index, VarTD, Λ+), we determined deviations of taxonomic structure of the studied mycological communities from the level of mean expected values, calculated based on the list of species, taking into account their systematic positions. The lowest values of index Δ+ were determined for communities on shells of M. galloprovincialis, 0.3 m depth, driftwood, stones and concrete. These communities had uneven distribution of species according to higher taxonomic ranks and minimum number of the highest taxa: 4–6 classes, 1–2 divisions, Fungi kingdom. Disproportion in species composition with decrease in the number of the highest taxa occurred in extreme environmental conditions. Using index Λ+, we found that the most complex taxonomic structure of fungi communities has developed on concrete and shells of C. gigas. In mycological communities on those substrates, the number of species was low (25 and 46), but they belonged to 4–7 classes, 2–3 divisions, 1–2 kingdoms. To compare the structures of mycological communities that have developed in such substrates in biotopes sea, sea-land-air, land-air, we compiled a list of fungi based on the literature data, which, taking into account our data, comprised 445 species of 240 genera, 103 families, 51 orders, 15 classes, 5 divisions, 2 kingdoms. The analysis revealed that on substrates with similar chemical composition, in all the biotopes, the species of the same divisions dominated (genus and family may vary). Therefore, in the biotope land-air – Hypocreales, Pleosporales, Eurotiales (genera Acremonium, Fusarium, Alternaria, Aspergillus, Penicillium); sea – Pleosporales, Eurotiales, Microascales (Alternaria, Aspergillus, Penicillium, Corollospora); sea-land-air – Pleosporales, Microascales (Alternaria, Leptosphaeria, Aspergillus, Penicillium, Corollospora, Halosarpheia). Monitoring of species composition of myxomycetes is needed in farms that cultivate industrial objects, recreation sites, various buildings for prevention of mycotoxin intoxication and infestation by mycodermatoses and other diseases caused by opportunistic and pathogenic fungi.

The mycological social network a way forward for conservation of fungal biodiversity

SummaryBecause knowledge of fungal diversity is very incomplete, it is possible that anthropogenic impacts are driving species to extinction before they have been discovered. Fungal inventories are still incomplete and do not reflect the complete diversity of this large taxon. Whilst molecular advancements are leading to an increased rate of species discovery, there is still much to be done to understand the diversity of fungi, identify rare species and establish conservation goals. Citizen science via social media could play an increasingly important role in mycological research, and its continued development should be supported and encouraged. The involvement of non-professionals in data collection helps increase public awareness, as well as extending the scope and efficiency of fungal surveys. Future academic mycological research could benefit from social media interaction and engagement with the amateur mycological community, which may accelerate the achievement of more effective conservation goals.

Provincial mycology and the legacy of Henry Thomas Soppitt (1858–1899) (W. T. Stearn Prize 2019)

The essay explores the life and legacy of Henry Thomas Soppitt, focusing on his elucidation of the life cycle of the rust fungus Puccinia bistortae (F. Strauss) DC. It focuses on the impact this discovery had on the Yorkshire mycological community during a period of threat to its standing caused by the professionalization of science and subsequent degradation of the amateur. It goes on to explores how two Yorkshire mycologists and participants in the discovery, Charles Crossland and James Needham, used accounts of Soppitt's discovery to ensure his legacy and, later, to attempt to ensure the legacy of their community. It shows how these accounts were varied in their content and medium and analyses the relative value of collections to printed text to Yorkshire mycologists through the differences in their accounts.

Review of Potential Pseudomonas Weaponry, Relevant to the Pseudomonas-Aspergillus Interplay, for the Mycology Community.

Pseudomonas aeruginosa is one of the most prominent opportunistic bacteria in airways of cystic fibrosis patients and in immunocompromised patients. These bacteria share the same polymicrobial niche with other microbes, such as the opportunistic fungus Aspergillus fumigatus. Their inter-kingdom interactions and diverse exchange of secreted metabolites are responsible for how they both fare in competition for ecological niches. The outcomes of their contests likely determine persistent damage and degeneration of lung function. With a myriad of virulence factors and metabolites of promising antifungal activity, P. aeruginosa products or their derivatives may prove useful in prophylaxis and therapy against A. fumigatus. Quorum sensing underlies the primary virulence strategy of P. aeruginosa, which serves as cell–cell communication and ultimately leads to the production of multiple virulence factors. Understanding the quorum-sensing-related pathogenic mechanisms of P. aeruginosa is a first step for understanding intermicrobial competition. In this review, we provide a basic overview of some of the central virulence factors of P. aeruginosa that are regulated by quorum-sensing response pathways and briefly discuss the hitherto known antifungal properties of these virulence factors. This review also addresses the role of the bacterial secretion machinery regarding virulence factor secretion and maintenance of cell–cell communication.

Virulence Factors in Sporothrix schenckii, One of the Causative Agents of Sporotrichosis

Sporothrix schenckii is one of the etiological agents of sporotrichosis, a fungal infection distributed worldwide. Both, the causative organism and the disease have currently received limited attention by the medical mycology community, most likely because of the low mortality rates associated with it. Nonetheless, morbidity is high in endemic regions and the versatility of S. schenckii to cause zoonosis and sapronosis has attracted attention. Thus far, virulence factors associated with this organism are poorly described. Here, comparing the S. schenckii genome sequence with other medically relevant fungi, genes involved in morphological change, cell wall synthesis, immune evasion, thermotolerance, adhesion, biofilm formation, melanin production, nutrient uptake, response to stress, extracellular vesicle formation, and toxin production are predicted and discussed as putative virulence factors in S. schenckii.

Candida auris: A Decade of Understanding of an Enigmatic Pathogenic Yeast.

Candida auris is an enigmatic yeast that continues to stimulate interest within the mycology community due its rapid and simultaneous emergence of distinct clades. In the last decade, almost 400 manuscripts have contributed to our understanding of this pathogenic yeast. With dynamic epidemiology, elevated resistance levels and an indication of conserved and unique pathogenic traits, it is unsurprising that it continues to cause clinical concern. This mini-review aims to summarise some of the key attributes of this remarkable pathogenic yeast.

Onygenalean Fungi as Major Human and Animal Pathogens

During the last two decades, many onygenalean pathogens were discovered, redefined, or re-classified from existing taxa into clusters of micro-species, among which the original genotypes often appeared to be uncommon and exceptional. The impact of these developments on the diagnosis and treatment of fungal diseases remains to be determined in most instances. This exciting collection of invited articles provides a full flavor of ongoing changes in the knowledge about taxonomy, genetics, ecology, epidemiology, and clinical spectra of human and animal pathogens classified in the order Onygenales. Recent developments have set the stage for an ambitious translational research agenda. Diagnostic mycology laboratories now need MALDI-TOF-MS spectra, PCR probes, and other specific tools to assist them in the rapid diagnosis of new species. Similarly, an educational set of type materials of new species needs to be readily available for enhanced expertise among the wider medical mycology community. As several new species were discovered retrospectively, it is crucial to expand the re-sampling to other fungal culture collections and archived paraffin tissues. Finally, clinical and laboratory investigations are needed to get an accurate assessment of the prevalence and impact of new pathogens as the cause of major fungal diseases.

Combined Antifungal Resistance and Biofilm Tolerance: the Global Threat of Candida auris.

The enigmatic yeast Candida auris has emerged over the last decade and rapidly penetrated our consciousness. The global threat from this multidrug-resistant yeast has generated a call to arms from within the medical mycology community. Over the past decade, our understanding of how this yeast has spread globally, its clinical importance, and how it tolerates and resists antifungal agents has expanded. This review highlights the clinical importance of antifungal resistance in C. auris and explores our current understanding of the mechanisms associated with azole, polyene, and echinocandin resistance. We also discuss the impact of phenotypic tolerance, with particular emphasis on biofilm-mediated resistance, and present new pipelines of antifungal drugs that promise new hope in the management of C. auris infection.

Vaginal microbiota of asymptomatic bacterial vaginosis and vulvovaginal candidiasis: Are they different from normal microbiota?

Vaginal microbiota contributes in maintaining and protecting the urogenital niche from infections and their sequelae. Despite extensive research, microbiome studies have often ignored asymptomatic bacterial vaginosis (BV) and vulvovaginal candidiasis (VVC). The present study aimed to explore the cultivable vaginal bacterial and mycological communities in women asymptomatic for BV and VVC using multiplex PCR and species-specific PCR. Vaginal swabs collected from 199 participants asymptomatic for urogenital infections, scored by Nugent criteria indicated 73.9% had normal microbiota, 11.6% intermediate and 14.5% BV. The most frequent Lactobacillus species in normal women were L. iners (69.4%), L. crispatus (24.5%), L. reuteri (20.4%). Women with BV colonized L. iners (62.1%); L. rhamnosus (41.4%); L. salivarius (13.7%) and L. reuteri (7.2%). Furthermore, L. crispatus was associated with normal microbiota, whereas L. iners was a frequent member of normal and dysbiotic microbiota. Lactobacillus abundance and species richness reduced in asymptomatic BV. Also L. crispatus, L. fermentum, L. acidophilus and L. delbruckii were absent in these women. L. iners significantly co-existed with other Lactobacillus species, indicating its failure in independently maintaining the healthy vaginal niche. Of 30.4% women detected with Candida, 72.1% constituted non-albicans Candida. Predominance of C. albicans increased from 18.4% in healthy to 60% in women with asymptomatic BV; whereas distribution of BV related bacteria did not vary across the groups. Heterogeneous population of lactobacilli in 80.8% of normal women calls attention towards cumulative effects of these species in safeguarding the vaginal microenvironment. Since the microbiota of asymptomatic BV was different from healthy, screening and management could be encouraged to avoid further complications of infections.

Correction to: Filamentous fungi associated with Brazilian stone samples: structure of the fungal community, diversity indexes, and ecological analysis

The present study aims to explore the diversity patterns of cultivable filamentous fungi and establish the ecological interactions that occur within the mycological community on soapstone in two distinct places: Architectural Complex of Caraca and Congonhas town, in Minas Gerais state, Brazil. A total of 200 isolates of filamentous fungi were detected and 72 distinct taxa were identified using molecular tools. The rock-inhabiting fungal communities were comprised of cosmopolitan taxa, which displayed high diversity and richness, but low dominance indexes at two of the sampling sites over the course of 12 months. The detection of organic acids secreted by the isolated filamentous fungal species revealed the potential for deterioration of the model stone by these fungi. Our results suggest that the isolated fungal communities of the two Brazilian localities have the ability to adapt to environmental adversities over the four seasons of the year, predicting adaptive population dynamics in the stone colonization. We conclude that this study on the ecological dynamics of the rock-inhabiting fungal communities can help to understand the competitive interactions between the environmental fungi. In addition, these fungi may contribute to the preservation of the Brazilian historical heritage that is vulnerable to environmental conditions.

Database establishment for the secondary fungal DNA barcode translational elongation factor 1α (TEF1α) 1.

With new or emerging fungal infections, human and animal fungal pathogens are a growing threat worldwide. Current diagnostic tools are slow, non-specific at the species and subspecies levels, and require specific morphological expertise to accurately identify pathogens from pure cultures. DNA barcodes are easily amplified, universal, short species-specific DNA sequences, which enable rapid identification by comparison with a well-curated reference sequence collection. The primary fungal DNA barcode, ITS region, was introduced in 2012 and is now routinely used in diagnostic laboratories. However, the ITS region only accurately identifies around 75% of all medically relevant fungal species, which has prompted the development of a secondary barcode to increase the resolution power and suitability of DNA barcoding for fungal disease diagnostics. The translational elongation factor 1α (TEF1α) was selected in 2015 as a secondary fungal DNA barcode, but it has not been implemented into practice, due to the absence of a reference database. Here, we have established a quality-controlled reference database for the secondary barcode that together with the ISHAM-ITS database, forms the ISHAM barcode database, available online at http://its.mycologylab.org/ . We encourage the mycology community for active contributions.

Can we use environmental DNA as holotypes?

The advantages and disadvantages of giving a valid name to a sequence of DNA detected from environmental specimens is presently a hot debate amongst the mycological community. The idea of using intracellular DNA (“mgDNA”) from environmental samples as holotypes seems at face value, to be a good idea, considering the expansion of knowledge among these ‘dark taxa’ or ‘dark matter fungi’ that it could provide (i.e. sequence based taxa without physical specimens and formal nomenclature). However, the limitations of using mgDNA as holotypes needs careful thought, i.e. can we use a short mgDNA fragment, which may contain a small amount of genetic information, to allow discrimination between species? What is the point and are the potential problems of giving valid scientific names to mgDNA? Numerous mycologists and taxonomists, who have many years of experience working on the taxonomy and phylogeny of different groups of fungi, are concerned about the consequences of providing valid names to mgDNA. There has been much debate, through several publications on the considerable problems of using mgDNA as holotypes. The proponents have tried to debate the virtues of using mgDNA as holotypes. Those against have shown that identification to species using mgDNA does not work in many fungal groups, while those for have shown cases where species can be identified with mgDNA. Different disciplines have different reasons and opinions for using mgDNA as holotypes, however even groups of the same disciplines have dissimilar ideas. In this paper we explore the use of mgDNA as holotypes. We provide evidences and opinions as to the use of mgDNA as holotypes from our own experiences. In no way do we attempt to degrade the study of DNA from environmental samples and the expansion of knowledge in to the dark taxa, but relate the issues to fungal taxonomy. In fact we show the value of using sequence data from these approaches, in dealing with the discovery of already named taxa, taxa numbers and ecological roles. We discuss the advantages and the pitfalls of using mgDNA from environmental samples as holotypes. The impacts of expanding the nomenclatural concept to allow using mgDNA from environmental samples as holotypes are also discussed. We provide evidence from case studies on Botryosphaeria, Colletotrichum, Penicillium and Xylaria. The case studies show that we cannot use mgDNA due to their short fragments and the fact that most ITS sequence data presently result from environmental sequencing. We conclude from the evidence that it is highly undesirable to use mgDNA as holotypes in naming fungal species. If this approach adopted, it would result in numerous problems where species identification cannot be confirmed due to limited sequence data available for the holotypes. We also propose an alternative DNA-based system for naming DNA based species which would provide considerably less problems and should be adopted.

Formal description of sequence-based voucherless Fungi: promises and pitfalls, and how to resolve them

There is urgent need for a formal nomenclature of sequence-based, voucherless Fungi, given that environmental sequencing has accumulated more than one billion fungal ITS reads in the Sequence Read Archive, about 1,000 times as many as fungal ITS sequences in GenBank. These unnamed Fungi could help to bridge the gap between 115,000 to 140,000 currently accepted and 2.2 to 3.8 million predicted species, a gap that cannot realistically be filled using specimen or culture-based inventories. The Code never aimed at placing restrictions on the nature of characters chosen for taxonomy, and the requirement for physical types is now becoming a constraint on the advancement of science. We elaborate on the promises and pitfalls of sequence-based nomenclature and provide potential solutions to major concerns of the mycological community. Types of sequence-based taxa, which by default lack a physical specimen or culture, could be designated in four alternative ways: (1) the underlying sample (’bag’ type), (2) the DNA extract, (3) fluorescent in situ hybridization (FISH), or (4) the type sequence itself. Only (4) would require changes to the Code and the latter would be the most straightforward approach, complying with three of the five principal functions of types better than physical specimens. A fifth way, representation of the sequence in an illustration, has been ruled as unacceptable in the Code. Potential flaws in sequence data are analogous to flaws in physical types, and artifacts are manageable if a stringent analytical approach is applied. Conceptual errors such as homoplasy intragenomic variation, gene duplication, hybridization, and horizontal gene transfer, apply to all molecular approaches and cannot be used as a specific argument against sequence-based nomenclature. The potential impact of these phenomena is manageable, as phylogenetic species delimitation has worked satisfactorily in Fungi. The most serious shortcoming of sequence-based nomenclature is the likelihood of parallel classifications, either by describing taxa that already have names based on physical types. or by using different markers to delimit species within the same lineage. The probability of inadvertently establishing sequence-based species that have names available is between 20.4 % and 1.5 % depending on the number of globally predicted fungal species. This compares favourably to a historical error rate of about 30 % based on physical types, and this rate could be reduced to practically zero by adding specific provisions to this approach in the Code. To avoid parallel classifications based on different markers, sequence-based nomenclature should be limited to a single marker, preferably the fungal ITS barcoding marker; this is possible since sequence-based nomenclature does not aim at accurate species delimitation but at naming lineages to generate a reference database, independent of whether these lineages represent species, closely related species complexes, or infraspecies. We argue that clustering methods are inappropriate for sequence-based nomenclature; this approach must instead use phylogenetic methods based on multiple alignments, combined with quantitative species recognition methods. We outline strategies to obtain higher-level phylogenies for ITS-based, voucherless species, including phylogenetic binning, ‘hijacking’ species delimitation methods, and temporal banding. We conclude that voucherless, sequence-based nomenclature is not a threat to specimen and culture-based fungal taxonomy, but a complementary approach capable of substantially closing the gap between known and predicted fungal diversity, an approach that requires careful work and high skill levels.

Identification of Batrachochytrium dendrobatidis inhibiting isolates via challenge against salamander skin microbiota.

Rapid declines in global amphibian populations can be partially attributed to chytridiomycosis caused by the fungus Batrachochytrium dendrobatidis (Bd). Due to how quickly it has spread worldwide, and the number of amphibians affected, this fungus is of increasing concern in both the herpetological and mycological communities. We characterized the cutaneous microbiome of Appalachian salamanders, which persist in the presence of the fungus despite high rates of infection in co‐occurring frog populations. The cutaneous microbiome of vertebrates is known as the first line of defense against invading pathogens. We hypothesized that these salamanders might have resident bacteria on their skin with the ability to inhibit Bd growth. Our objectives were to 1) isolate resident bacteria from salamander skin swabs, 2) challenge these isolates against Bd, and 3) genotype all inhibitory isolates. During the summer of 2016, 82 swabs were collected from three salamander genera including Desmognathus (n=36), Eurycea (n=20), and Plethodon (n=26). To isolate cutaneous bacteria, swabs were streaked on Reasoner's 2A agar plates that we checked daily for two weeks. Colonies with unique morphology were isolated onto secondary plates until pure cultures were obtained. To challenge the isolates against Bd, zoospores were harvested from cultures that had been aged 5–7 days, counted with a hemocytometer, and diluted to approximately 1,000,000 cells/milliliter (mL). Tryptone agar plates were then flooded with 1.0 mL of standardized zoospore solution and dried for 45 minutes in a laminar flow hood. Bacterial isolates were streaked onto each plate in co‐culture with Bd. Plates were incubated at room temperature for four days, examined for noticeable zones of inhibition (ZOI), measured in millimeters. Isolates that produced a measurable ZOI were photographed and will be screened in further testing against Bd. We used direct colony PCR and Sanger sequencing to identify all inhibitory isolates. Presently, 63 of the 472 acquired isolates have been challenged, 16 of which produced a ZOI. The challenge phase is ongoing and we anticipate discovery of additional candidate probiotic species. In addition, we plan to test for possible synergistic inhibition by challenging candidates in polyculture.Support or Funding InformationGreat Smokey Mountains Conservation Association and Tennessee Herpetological SocietyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

New light on names and naming of dark taxa.

A growing proportion of fungal species and lineages are known only from sequence data and cannot be linked to any physical specimen or resolved taxonomic name. Such fungi are often referred to as “dark taxa” or “dark matter fungi”. As they lack a taxonomic identity in the form of a name, they are regularly ignored in many important contexts, for example in legalisation and species counts. It is therefore very urgent to find a system to also deal with these fungi. Here, issues relating to the taxonomy and nomenclature of dark taxa are discussed and a number of questions that the mycological community needs to consider before deciding on what system/s to implement are highlighted.

Green digitization: Online botanical collections data answering real‐world questions

Green digitization: Online botanical collections data answering real‐world questions