Ethanol and 2-phenylethanol production by bee-isolated Meyerozyma caribbica strains.

Investigating the biotechnological potential of wild microorganisms is paramount for optimizing bioprocesses. Given this premise, we looked for yeasts in Brazilian native stingless bees, considering the recognized potential of pollinating insect-associated microorganisms for the production of volatile organic compounds (VOCs). Two yeast strains of the species Meyerozyma caribbica were isolated from bees Scaptotrigona postica and evaluated for their fermentative capacity. Both yeasts were capable of fermenting sucrose (the main sugar used in the Brazilian ethanol industry) with over 90% efficiency and yields of up to 0.504 g/g. Through an experimental design analysis (CCD), it was verified that the ethanol productivity of these yeasts can also benefit from high concentrations of sucrose and low pH values, desirable traits for microorganisms in this biofuel production. At the same time, CCD analyses also showed the great capacity of these M. caribbica strains to produce another alcohol of broad biotechnological interest, 2-phenylethanol. Interestingly, the statistical analyses demonstrated that greater production of this compound can occur at high sugar concentrations and low availability of nitrogen sources, which can be easily achieved using residual low-cost feedstocks. Thus, our results suggest that these M. caribbica strains may be efficiently used in both ethanol and 2-phenylethanol production.

Comparative analysis of hydrophobicity and dentin adhesion ability in Candida albicans strains.

OBJECTIVE: Adhesion to dentin is a first step for a successful microbial root canal colonization. Cell hydrophobicity seems to have some influence in the Candida species adhesion to surfaces. To measure cell surface hydrophobicity and to investigate the adherence ability to human dentin among Candida albicans strains isolated from root canal and lingual dorsum via an in vitro study. METHODOLOGY: adhesion was quantified in function of dentin area covered by blastospores and/or hyphae presence detected by epifluorescence microscope. Cell surface hydrophobicity was estimated by assessing the percentage migration of cells from an aqueous phase to a hydrocarbon phase. Contact angles were measured by the sessile drop technique on the dentin surface using a contact angle measurements apparatus. We also examined the correlation between adhesion ability and hydrophobicity. RESULTS: although there was some intra-species variation in cell surface hydrophobicity, most isolates were characterized by moderate hydrophobicity. There was no significant difference in this parameter when the isolation niche was considered. Both root canal and lingual dorsum yeasts were able to adhere to dentin. No association was found between the strains' site of isolation and adhesion. Moreover, cell surface hydrophobicity and adhesion ability were not correlated. CONCLUSION: although hydrophobicity can influence Candida albicans virulence in many ways, this study suggests that this parameter by itself was not a good predictor of adhesion to dentin.

Enzymes and biosurfactants of industrial interest produced by culturable fungi present in sediments of Boeckella Lake, Hope Bay, north-east Antarctic Peninsula.

This study characterized cultivable fungi present in sediments obtained from Boeckella Lake, Hope Bay, in the north-east of the Antarctic Peninsula, and evaluated their production of enzymes and biosurfactants of potential industrial interest. A total of 116 fungal isolates were obtained, which were classified into 16 genera within the phyla Ascomycota, Basidiomycota and Mortierellomycota, in rank. The most abundant genera of filamentous fungi included Pseudogymnoascus, Pseudeurotium and Antarctomyces; for yeasts, Thelebolales and Naganishia taxa were dominant. Overall, the lake sediments exhibited high fungal diversity and moderate richness and dominance. The enzymes esterase, cellulase and protease were the most abundantly produced by these fungi. Ramgea cf. ozimecii, Holtermanniella wattica, Leucosporidium creatinivorum, Leucosporidium sp., Mrakia blollopis, Naganishia sp. and Phenoliferia sp. displayed enzymatic index > 2. Fourteen isolates of filamentous fungi demonstrated an Emulsification Index 24% (EI24%) ≥ 50%; among them, three isolates of A. psychrotrophicus showed an EI24% > 80%. Boeckella Lake itself is in the process of drying out due to the impact of regional climate change, and may be lost completely in approaching decades, therefore hosts a threatened community of cultivable fungi that produce important biomolecules with potential application in biotechnological processes.

Diversity and enzymatic, biosurfactant and phytotoxic activities of culturable Ascomycota fungi present in marine sediments obtained near the South Shetland Islands, maritime Antarctica.

We studied the culturable fungal community recovered from deep marine sediments in the maritime Antarctic, and assessed their capabilities to produce exoenzymes, emulsifiers and metabolites with phytotoxic activity. Sixty-eight Ascomycota fungal isolates were recovered and identified. The most abundant taxon recovered was the yeast Meyerozyma guilliermondii, followed by the filamentous fungi Penicillium chrysogenum, P. cf. palitans, Pseudeurotium cf. bakeri, Thelebolus balaustiformis, Antarctomyces psychrotrophicus and Cladosporium sp. Diversity indices displayed low values overall, with the highest values obtained at shallow depth, decreasing to the deepest location sampled. Only M. guilliermondii and P. cf. palitans were detected in the sediments at all depths sampled, and were the most abundant taxa at all sample sites. The most abundant enzymes detected were proteases, followed by invertases, cellulases, lipases, carrageenases, agarases, pectinases and esterases. Four isolates showed good biosurfactant activity, particularly the endemic species A. psychrotrophicus. Twenty-four isolates of P. cf. palitans displayed strong phytotoxic activities against the models Lactuca sativa and Allium schoenoprasum. The cultivable fungi recovered demonstrated good biosynthetic activity in the production of hydrolytic exoenzymes, biosurfactant molecules and metabolites with phytotoxic activity, reinforcing the importance of documenting the taxonomic, ecological and biotechnological properties of fungi present in deep oceanic sediments of the Southern Ocean.

Comparative analysis of hydrophobicity and dentin adhesion ability in Candida albicans strains

Abstract Adhesion to dentin is a first step for a successful microbial root canal colonization. Cell hydrophobicity seems to have some influence in the Candida species adhesion to surfaces. Objective to measure cell surface hydrophobicity and to investigate the adherence ability to human dentin among Candida albicans strains isolated from root canal and lingual dorsum via an in vitro study. Methodology adhesion was quantified in function of dentin area covered by blastospores and/or hyphae presence detected by epifluorescence microscope. Cell surface hydrophobicity was estimated by assessing the percentage migration of cells from an aqueous phase to a hydrocarbon phase. Contact angles were measured by the sessile drop technique on the dentin surface using a contact angle measurements apparatus. We also examined the correlation between adhesion ability and hydrophobicity. Results although there was some intra-species variation in cell surface hydrophobicity, most isolates were characterized by moderate hydrophobicity. There was no significant difference in this parameter when the isolation niche was considered. Both root canal and lingual dorsum yeasts were able to adhere to dentin. No association was found between the strains' site of isolation and adhesion. Moreover, cell surface hydrophobicity and adhesion ability were not correlated. Conclusion although hydrophobicity can influence Candida albicans virulence in many ways, this study suggests that this parameter by itself was not a good predictor of adhesion to dentin.

Further evidences of an emerging stingless bee-yeast symbiosis.

Symbiotic interactions between microorganisms and social insects have been described as crucial for the maintenance of these multitrophic systems, as observed for the stingless bee Scaptotrigona depilis and the yeast Zygosaccharomyces sp. SDBC30G1. The larvae of S. depilis ingest fungal filaments of Zygosaccharomyces sp. SDBC30G1 to obtain ergosterol, which is the precursor for the biosynthesis of ecdysteroids that modulate insect metamorphosis. In this work, we find a similar insect-microbe interaction in other species of stingless bees. We analyzed brood cell samples from 19 species of stingless bees collected in Brazil. The osmophilic yeast Zygosaccharomyces spp. was isolated from eight bee species, namely Scaptotrigona bipunctata, S. postica, S. tubiba, Tetragona clavipes, Melipona quadrifasciata, M. fasciculata, M. bicolor, and Partamona helleri. These yeasts form pseudohyphae and also accumulate ergosterol in lipid droplets, similar to the pattern observed for S. depilis. The phylogenetic analyses including various Zygosaccharomyces revealed that strains isolated from the brood cells formed a branch separated from the previously described Zygosaccharomyces species, suggesting that they are new species of this genus and reinforcing the symbiotic interaction with the host insects.

Aspergillus sclerotiorum lipolytic activity and its application in bioremediation of high-fat dairy wastewater environments.

Oils and grease (O&G) have low affinity for water and represent a class of pollutants present in the dairy industry. Enzyme-mediated bioremediation using biocatalysts, such as lipases, has shown promising potential in biotechnology, as they are versatile catalysts with high enantioselectivity and regioselectivity and easy availability, being considered a clean technology (white biotechnology). Specially in the treatment of effluents from dairy industries, these enzymes are of particular importance as they specifically hydrolyze O&G. In this context, the objective of this work is to prospect filamentous fungi with the ability to synthesize lipases for application in a high-fat dairy wastewater environment. We identified and characterized the fungal species Aspergillus sclerotiorum as a good lipase producer. Specifically, we observed highest lipolytic activity (20.72 U g-1) after 96 h of fermentation using sunflower seed as substrate. The fungal solid fermented was used in the bioremediation in dairy effluent to reduce O&G. The experiment was done in kinetic from 24 to 168 h and reduced over 90% of the O&G present in the sample after 168 h. Collectively, our work demonstrated the efficiency and applicability of fungal fermented solids in bioremediation and how this process can contribute to a more sustainable wastewater pretreatment, reducing the generation of effluents produced by dairy industries.

Communities of culturable freshwater fungi present in Antarctic lakes and detection of their low-temperature-active enzymes.

We evaluated the diversity and enzymatic activities of culturable fungi recovered from cotton baits submerged for 2 years in Hennequin Lake, King George Island, and from benthic biofilms in Kroner Lake, Deception Island, South Shetland Islands, maritime Antarctica. A total of 154 fungal isolates were obtained, representing in rank abundance the phyla Ascomycota, Basidiomycota and Mortierellomycota. Thelebolus globosus, Goffeauzyma sp., Pseudogymnoascus verrucosus and Metschnikowia australis were the most abundant taxa. The fungal community obtained from the biofilm was more diverse and richer than that recovered from the cotton baits. However, diversity indices suggested that the lakes may harbour further fungal diversity. The capabilities of all cultured fungi to produce the extracellular enzymes cellulase, protease, lipase, agarase, carrageenase, invertase, amylase, esterase, pectinase, inulinase and gelatinase at low temperature were evaluated. All enzymes were detected, but the most widely produced were protease and pectinase. The best enzymatic indices were obtained from Holtermanniella wattica (for invertase, esterase), Goffeauzyma sp. (amylase), Metschnikowia australis (protease), Mrakia blollopis (cellulase, pectinase), Pseudogymnoascus verrucosus (agarase, carrageenase) and Leucosporidium fragarium (inulinase). The detection of multiple enzymes reinforces the ecological role of fungi in nutrient cycling in Antarctic lakes, making nutrients available to the complex aquatic food web. Furthermore, such low-temperature-active enzymes may find application in different biotechnological processes, such as in the textile, pharmaceutical, food, detergent and paper industries, as well as environmental application in pollutant bioremediation processes.

Does maritime Antarctic permafrost harbor environmental fungi with pathogenic potential?

We assessed the potentially pathogenic fungi present in Antarctic permafrost and the overlying active layer on King George, Robert, Livingston and Deception Islands in the South Shetland Islands archipelago, maritime Antarctica. Permafrost and active layer sub-samples were incubated at 37 °C to select fungi able to grow inside the human body. A total of 67 fungal isolates were obtained, 27 from the permafrost and 40 from the active layer. These represented 18 taxa of the genera Alternaria, Aspergillus, Curvularia, Penicillium, Rhodotorula and Talaromyces. The majority of fungi detected occurred exclusively either in the permafrost or the active layer at each site. Only Aspergillus thermomutatus, Penicillium cf. chrysogenum and Rhodotorula cf. mucilaginosa were present in both permafrost and active layer samples from the same site. The yeast R. cf. mucilaginosa was recovered from both in at least two sites. The genus Penicillium was the most abundant and widely distributed genus in both permafrost and active layer samples across the sites sampled. All fungal isolates were screened using enzymatic, pH and antifungal assays to identify their virulence potential. Aspergillus hiratsukae, A. thermomutatus and R. cf. mucilaginosa, known human opportunistic fungi, were identified, displayed phospholipase, esterase, proteinase and hemolytic activities. All three also displayed the ability to grow at 40°, 45° and/or 50 °C and resistance to fluconazole and itraconazole; additionally, R. cf. mucilaginosa showed resistance to amphotericin B and viability after 100 d at -80 °C. A. thermomutatus UFMGCB 17415 killed the entire larvae of Tenebrio molitor in six days and R. cf. mucilaginosa UFMGCB 17448 and 17473 in three and four days, respectively. The melting of maritime Antarctic permafrost as a result of climate change may threaten the release of wild strains of pathogenic fungi geographically isolated for long time, which may in turn be transported within and beyond Antarctica by different biological and non-biological vectors.

Diversity, distribution and ecology of fungal communities present in Antarctic lake sediments uncovered by DNA metabarcoding.

We assessed fungal diversity in sediments obtained from four lakes in the South Shetland Islands and James Ross Island, Antarctica, using DNA metabarcoding. We detected 218 amplicon sequence variants (ASVs) dominated by the phyla Ascomycota, Basidiomycota, Mortierellomycota, Mucoromycota and Chytridiomycota. In addition, the rare phyla Aphelidiomycota, Basidiobolomycota, Blastocladiomycota, Monoblepharomycota, Rozellomycota and Zoopagomycota as well as fungal-like Straminopila belonging to the phyla Bacillariophyta and Oomycota were detected. The fungal assemblages were dominated by unknown fungal taxa (Fungal sp. 1 and Fungal sp. 2), followed by Talaromyces rubicundus and Dactylonectria anthuriicola. In general, they displayed high diversity, richness and moderate dominance. Sequences representing saprophytic, pathogenic and symbiotic fungi were detected, including the phytopathogenic fungus D. anthuriicola that was abundant, in the relatively young Soto Lake on Deception Island. The lake sediments studied contained the DNA of rich, diverse and complex fungal communities, including both fungi commonly reported in Antarctica and other taxa considered to be rare. However, as the study was based on the use of environmental DNA, which does not unequivocally confirm the presence of active or viable organisms, further studies using other approaches such as shotgun sequencing are required to elucidate the ecology of fungi in these Antarctic lake sediments.

Fungal diversity in a sediment core from climate change impacted Boeckella Lake, Hope Bay, north-eastern Antarctic Peninsula assessed using metabarcoding.

We studied the fungal DNA present in a lake sediment core obtained from Trinity Peninsula, Hope Bay, north-eastern Antarctic Peninsula, using metabarcoding through high-throughput sequencing (HTS). Sequences obtained were assigned to 146 amplicon sequence variants (ASVs) primarily representing unknown fungi, followed by the phyla Ascomycota, Rozellomycota, Basidiomycota, Chytridiomycota and Mortierellomycota. The most abundant taxa were assigned to Fungal sp., Pseudeurotium hygrophilum, Rozellomycota sp. 1, Pseudeurotiaceae sp. 1 and Chytridiomycota sp. 1. The majority of the DNA reads, representing 40 ASVs, could only be assigned at higher taxonomic levels and may represent taxa not currently included in the sequence databases consulted and/or be previously undescribed fungi. Different sections of the core were characterized by high sequence diversity, richness and moderate ecological dominance indices. The assigned diversity was dominated by cosmopolitan cold-adapted fungi, including known saprotrophic, plant and animal pathogenic and symbiotic taxa. Despite the overall dominance of Ascomycota and Basidiomycota and psychrophilic Mortierellomycota, members of the cryptic phyla Rozellomycota and Chytridiomycota were also detected in abundance. As Boeckella Lake may cease to exist in approaching decades due the effects of local climatic changes, it also an important location for the study of the impacts of these changes on Antarctic microbial diversity.

Physiological comparisons among Spathaspora passalidarum, Spathaspora arborariae, and Scheffersomyces stipitis reveal the bottlenecks for their use in the production of second-generation ethanol.

The microbial conversion of pentoses to ethanol is one of the major drawbacks that limits the complete use of lignocellulosic sugars. In this study, we compared the yeast species Spathaspora arborariae, Spathaspora passalidarum, and Sheffersomyces stipitis regarding their potential use for xylose fermentation. Herein, we evaluated the effects of xylose concentration, presence of glucose, and temperature on ethanol production. The inhibitory effects of furfural, hydroxymethylfurfural (HMF), acetic acid, and ethanol were also determined. The highest ethanol yield (0.44 g/g) and productivity (1.02 g/L.h) were obtained using Sp. passalidarum grown in 100 g/L xylose at 32 °C. The rate of xylose consumption was reduced in the presence of glucose for the species tested. Hydroxymethylfurfural did not inhibit the growth of yeasts, whereas furfural extended their lag phase. Acetic acid inhibited the growth and fermentation of all yeasts. Furthermore, we showed that these xylose-fermenting yeasts do not produce ethanol concentrations greater than 4% (v/v), probably due to the inhibitory effects of ethanol on yeast physiology. Our data confirm that among the studied yeasts, Sp. passalidarum is the most promising for xylose fermentation, and the low tolerance to ethanol is an important aspect to be improved to increase its performance for second-generation (2G) ethanol production. Our molecular data showed that this yeast failed to induce the expression of some classical genes involved in ethanol tolerance. These findings suggest that Sp. passalidarum may have not activated a proper response to the stress, impacting its ability to overcome the negative effects of ethanol on the cells.

Diversity of Fungi Present in Permafrost in the South Shetland Islands, Maritime Antarctic.

We assess the fungal diversity present in permafrost from different islands in the South Shetland Islands archipelago, maritime Antarctic, using next-generation sequencing (NGS). We detected 1,003,637 fungal DNA reads representing, in rank abundance order, the phyla Ascomycota, Mortierellomycota, Basidiomycota, Chytridiomycota, Rozellomycota, Mucoromycota, Calcarisporiellomycota and Zoopagomycota. Ten taxa were dominant these being, in order of abundance, Pseudogymnoascus appendiculatus, Penicillium sp., Pseudogymnoascus roseus, Penicillium herquei, Curvularia lunata, Leotiomycetes sp., Mortierella sp. 1, Mortierella fimbricystis, Fungal sp. 1 and Fungal sp. 2. A further 38 taxa had intermediate abundance and 345 were classified as rare. The total fungal community detected in the permafrost showed high indices of diversity, richness and dominance, although these varied between the sampling locations. The use of a metabarcoding approach revealed the presence of DNA of a complex fungal assemblage in the permafrost of the South Shetland Islands including taxa with a range of ecological functions among which were multiple animal, human and plant pathogenic fungi. Further studies are required to determine whether the taxa identified are present in the form of viable cells or propagules and which might be released from melting permafrost to other Antarctic habitats and potentially dispersed more widely.

Fungi in the Antarctic Cryosphere: Using DNA Metabarcoding to Reveal Fungal Diversity in Glacial Ice from the Antarctic Peninsula Region.

We assessed fungal diversity present in glacial from the Antarctic Peninsula using DNA metabarcoding through high-throughput sequencing (HTS). We detected a total of 353,879 fungal DNA reads, representing 94 genera and 184 taxa, in glacial ice fragments obtained from seven sites in the north-west Antarctic Peninsula and South Shetland Islands. The phylum Ascomycota dominated the sequence diversity, followed by Basidiomycota and Mortierellomycota. Penicillium sp., Cladosporium sp., Penicillium atrovenetum, Epicoccum nigrum, Pseudogymnoascus sp. 1, Pseudogymnoascus sp. 2, Phaeosphaeriaceae sp. and Xylaria grammica were the most dominant taxa, respectively. However, the majority of the fungal diversity comprised taxa of rare and intermediate relative abundance, predominately known mesophilic fungi. High indices of diversity and richness were calculated, along with moderate index of dominance, which varied among the different sampling sites. Only 26 (14%) of the total fungal taxa detected were present at all sampling sites. The identified diversity was dominated by saprophytic taxa, followed by known plant and animal pathogens and a low number of symbiotic fungi. Our data suggest that Antarctic glacial ice may represent a hotspot of previously unreported fungal diversity; however, further studies are required to integrate HTS and culture approaches to confirm viability of the taxa detected.

Ecological succession of fungal and bacterial communities in Antarctic mosses affected by a fairy ring disease.

We evaluated fungal and bacterial diversity in an established moss carpet on King George Island, Antarctica, affected by 'fairy ring' disease using metabarcoding. A total of 127 fungal and 706 bacterial taxa were assigned. Ascomycota dominated the fungal assemblages, followed by Basidiomycota, Rozellomycota, Chytridiomycota, Mortierellomycota and Monoblepharomycota. The fungal community displayed high indices of diversity, richness and dominance, which increased from healthy through infected to dead moss samples. A range of fungal taxa were more abundant in dead rather than healthy or fairy ring moss samples. Bacterial diversity and richness were greatest in healthy moss and least within the infected fairy ring. The dominant prokaryotic phyla were Actinobacteriota, Proteobacteria, Bacteroidota and Cyanobacteria. Cyanophyceae sp., whilst consistently dominant, were less abundant in fairy ring samples. Our data confirmed the presence and abundance of a range of plant pathogenic fungi, supporting the hypothesis that the disease is linked with multiple fungal taxa. Further studies are required to characterise the interactions between plant pathogenic fungi and their host Antarctic mosses. Monitoring the dynamics of mutualist, phytopathogenic and decomposer microorganisms associated with moss carpets may provide bioindicators of moss health.

Performance of xylose-fermenting yeasts in oat and soybean hulls hydrolysate and improvement of ethanol production using immobilized cell systems.

We investigated the fermentation of a mixture of oat and soybean hulls (1:1) subjected to acid (AH) or enzymatic (EH) hydrolyses, with both showing high osmotic pressures (> 1200 Osm kg-1) for the production of ethanol. Yeasts of genera Spathaspora, Scheffersomyces, Sugiymaella, and Candida, most of them biodiverse Brazilian isolates and previously untested in bioprocesses, were cultivated in these hydrolysates. Spathaspora passalidarum UFMG-CM-469 showed the best ethanol production kinetics in suspended cells cultures in acid hydrolysate, under microaerobic and anaerobic conditions. This strain was immobilized in LentiKats® (polyvinyl alcohol) and cultured in AH and EH. Supplementation of hydrolysates with crude yeast extract and peptone was also performed. The highest ethanol production was obtained using hydrolysates supplemented with crude yeast extract (AH-CYE and EH-CYE) showing yields of 0.40 and 0.44 g g-1, and productivities of 0.39 and 0.29 g (L h)-1, respectively. The reuse of the immobilized cells was tested in sequential fermentations of AH-CYE, EH-CYE, and a mixture of acid and enzymatic hydrolysates (AEH-CYE) operated under batch fluidized bed, with ethanol yields ranging from 0.31 to 0.40 g g-1 and productivities from 0.14 to 0.23 g (L h)-1. These results warrant further research using Spathaspora yeasts for second-generation ethanol production.

Microbial communities associated with honey bees in Brazil and in the United States.

Honey bee colony losses worldwide call for a more in-depth understanding of the pathogenic and mutualistic components of the honey bee microbiota and their relation with the environment. In this descriptive study, we characterized the yeast and bacterial communities that arise from six substrates associated with honey bees: corbicular pollen, beebread, hive debris, intestinal contents, body surface of nurses and forager bees, comparing two different landscapes, Minas Gerais, Brazil and Maryland, United States. The sampling of five hives in Brazil and four in the USA yielded 217 yeast and 284 bacterial isolates. Whereas the yeast community, accounted for 47 species from 29 genera, was dominated in Brazil by Aureobasidium sp. and Candida orthopsilosis, the major yeast recovered from the USA was Debaryomyces hansenii. The bacterial community was more diverse, encompassing 65 species distributed across 31 genera. Overall, most isolates belonged to Firmicutes, genus Bacillus. Among LAB, species from Lactobacillus were the most prevalent. Cluster analysis evidenced high structuration of the microbial communities, with two distinguished microbial groups between Brazil and the United States. In general, the higher difference among sites and substrates were dependents on the turnover effect (~ 93% of the beta diversity), with a more pronounced effect of nestedness (~ 28%) observed from Brazil microbiota change. The relative abundance of yeasts and bacteria also showed the dissimilarity of the microbial communities between both environments. These results provide a comprehensive view of microorganisms associated with A. mellifera, highlighting the importance of the environment in the establishment of the microbiota associated with honey bees.

Diversity of fungal DNA in lake sediments on Vega Island, north-east Antarctic Peninsula assessed using DNA metabarcoding.

We assessed the diversity of fungal DNA present in sediments of three lakes on Vega Island, north-east Antarctic Peninsula using metabarcoding through high-throughput sequencing (HTS). A total of 640,902 fungal DNA reads were detected, which were assigned to 224 taxa of the phyla Ascomycota, Rozellomycota, Basidiomycota, Chytridiomycota and Mortierellomycota, in rank order of abundance. The most abundant genera were Pseudogymnoascus, Penicillium and Mortierella. However, a majority (423,508, 66%) of the reads, representing by 43 ASVs, could only be assigned at higher taxonomic levels and may represent taxa not currently included in the sequence databases used or be new or previously unreported taxa present in Antarctic lakes. The three lakes were characterized by high sequence diversity, richness, and moderate dominance indices. The ASVs were dominated by psychrotolerant and cosmopolitan cold-adapted Ascomycota, Basidiomycota and Mortierellomycota commonly reported in Antarctic environments. However, other taxa detected included unidentified members of Rozellomycota and Chytridiomycota species not previously reported in Antarctic lakes. The assigned diversity was composed mainly of taxa recognized as decomposers and pathogens of plants and invertebrates.

Fungal diversity present on rocks from a polar desert in continental Antarctica assessed using DNA metabarcoding.

We evaluated the fungal diversity associated with carbonate veins and two types of salt encrustation in rocks in a polar desert region of continental Antarctica using DNA a metabarcoding approach. We detected 262,268 reads grouped into 517 amplicon sequence variants (ASVs) assigned to the phyla Ascomycota, Basidiomycota, Mortierellomycota and Mucoromycota. Fourteen ASVs belonging to the genera Trichosporon, Mortierella, Penicillium, Aspergillus, Cladosporium, Coprinellus, Pleurotus and Pseudogymnoascus were assessed to be dominant taxa. The fungal communities of the three habitats sampled displayed high diversity indices when compared with other habitats of Antarctica, although differing in detail, with the highest diversity indices in the gypsum crust type 2. Only 48 of the 517 ASVs (9.28%) were detected in all three habitats, including dominant, intermediate and minor components. In contrast with previous studies of fungal communities living in the ultra-extreme conditions of continental Antarctica, application of metabarcoding revealed the DNA of a rich and complex resident fungal community. The ASVs detected included fungi with different ecological roles, with xerophilic, human- and animal-associated, phytopathogenic, saprotrophic, mutualistic, psychrotolerant and cosmopolitan taxa. This sequence diversity may be the result of deposition of fungal propagules over time driven by air currents, precipitation or human activities in the region.

Fungi Present in Antarctic Deep-Sea Sediments Assessed Using DNA Metabarcoding.

We assessed fungal diversity in deep-sea sediments obtained from different depths in the Southern Ocean using the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA by metabarcoding through high-throughput sequencing (HTS). We detected 655,991 DNA reads representing 263 fungal amplicon sequence variants (ASVs), dominated by Ascomycota, Basidiomycota, Mortierellomycota, Mucoromycota, Chytridiomycota and Rozellomycota, confirming that deep-sea sediments can represent a hotspot of fungal diversity in Antarctica. The community diversity detected included 17 dominant fungal ASVs, 62 intermediate and 213 rare. The dominant fungi included taxa of Mortierella, Penicillium, Cladosporium, Pseudogymnoascus, Phaeosphaeria and Torula. Despite the extreme conditions of the Southern Ocean benthos, the total fungal community detected in these marine sediments displayed high indices of diversity and richness, and moderate dominance, which varied between the different depths sampled. The highest diversity indices were obtained in sediments from 550 m and 250 m depths. Only 49 ASVs (18.63%) were detected at all the depths sampled, while 16 ASVs were detected only in the deepest sediment sampled at 1463 m. Based on sequence identities, the fungal community included some globally distributed taxa, primarily recorded otherwise from terrestrial environments, suggesting transport from these to deep marine sediments. The assigned taxa included symbionts, decomposers and plant-, animal- and human-pathogenic fungi, suggesting that deep-sea sediments host a complex fungal diversity, although metabarcoding does not itself confirm that living or viable organisms are present.