Endolithic Fungal Diversity in Antarctic Oligocene Rock Samples Explored Using DNA Metabarcoding.

In this study, we evaluated the fungal diversity present associated with cores of Oligocene rocks using a DNA metabarcoding approach. We detected 940,969 DNA reads grouped into 198 amplicon sequence variants (ASVs) representing the phyla Ascomycota, Basidiomycota, Mortierellomycota, Chytridiomycota, Mucoromycota, Rozellomycota, Blastocladiomycota, Monoblepharomycota, Zoopagomycota, Aphelidiomycota (Fungi) and the fungal-like Oomycota (Stramenopila), in rank abundance order. Pseudogymnoascus pannorum, Penicillium sp., Aspergillus sp., Cladosporium sp., Aspergillaceae sp. and Diaporthaceae sp. were assessed to be dominant taxa, with 22 fungal ASVs displaying intermediate abundance and 170 being minor components of the assigned fungal diversity. The data obtained displayed high diversity indices, while rarefaction indicated that the majority of the diversity was detected. However, the diversity indices varied between the cores analysed. The endolithic fungal community detected using a metabarcoding approach in the Oligocene rock samples examined contains a rich and complex mycobiome comprising taxa with different lifestyles, comparable with the diversity reported in recent studies of a range of Antarctic habitats. Due to the high fungal diversity detected, our results suggest the necessity of further research to develop strategies to isolate these fungi in culture for evolutionary, physiological, and biogeochemical studies, and to assess their potential role in biotechnological applications.

Contribution by Giant petrels and Brown skuas to soil phosphatization in Harmony Point - Maritime Antarctica.

This research was conducted for the purpose to evaluate the contribution Giant petrels and Brown skuas nestings in the formation of ornithogenic soils by the phosphatization process in Harmony Point, Maritime Antarctic. Ten nests were selected to collect topsoil samples (0-20 cm), from 0 m up to 20 m away, with regular spacing of 2 m. The analysis of the samples included the physical, chemical, mineralogical and geochemical analyzes. Phosphate minerals were identified. The presence of high content of some trace elements, such as Zn, Cu and Sr is associated with the bird's excrements. Total-P and bioavailable-P recorded higher levels. This result demonstrates the importance of the time factor in the bird's nesting, as well as in the development of the soil in these soil-forming environment. Phosphatization in these areas is not restricted to the specific location of the nest, since high values of P have been identified at distances between 8 and 12 m, from de nest's top. This suggests the transport of P rich solutions and phosphatized material along fractures by the freeze-thaw cycles, contributing to increase the geographical expression of this phenomenon in this ice-free area, consequently the development of soils and the establishment of vegetation.

Soils under seal carcasses with varying degrees of decomposition: oasis of nutrients and vegetation in Antarctica.

Areas of high concentration of seal carcasses have been observed in localized areas of James Ross Island, Antarctica. Such carcasses show an unusual vegetation development, in a semi-arid area with bare soils under intense winds, high salinity and sandy texture. We investigated carcasses of seals around a lake in James Ross Island, with four different stages of decomposition, with three replicates: Seal (S01), with recently mummified carcasses; S02, with partially degraded carcasses; S03, with broken carcasses with partially degraded exposed bones, and S04, with completely broken, scattered skeletons. The vegetation showed a maximum degree of development in carcasses at stages S02 and S03, with the environment between the skin and the skeleton as the preferred place for vegetation establishment. The chemical alteration was greater with increasing carcass decomposition but reduced with the spreading and final decomposition of the bones, with anomalous values observed only in the vicinity of the carcasses. It is concluded that the presence of carcasses of seals, concentrated in wet places, even in a semi-desert climate, represent important oases of nutrients, with a combination of physical and chemical effects throughout the decomposition process that favor plant establishment and succession.

Spatial and Scientometric study of the Brazilian scientific production on Antarctic soils and permafrost.

This article carried out the first scientometric and spatial analysis of Brazilian scientific production on Antarctic soils and permafrost, based on all publications available from the Scopus and Web of Science databases. Information on co-authorship, citation, research topics, and sampling sites was used to understand the social and theoretical structure as well as the spatial dynamics of this research field in Brazil over the last 25 years. We highlight that Brazil is presently, the main country to study the soils and permafrost of Maritime Antarctica, in addition to having an international robust and prolific production, with high impact on the literature, and widely distributed throughout the studied region. It was also possible to identify potential future international partners, new research locations and strategic research themes.

Soil Fungal Diversity and Ecology Assessed Using DNA Metabarcoding along a Deglaciated Chronosequence at Clearwater Mesa, James Ross Island, Antarctic Peninsula.

We studied the fungal diversity present in soils sampled along a deglaciated chronosequence from para- to periglacial conditions on James Ross Island, north-east Antarctic Peninsula, using DNA metabarcoding. A total of 88 amplicon sequence variants (ASVs) were detected, dominated by the phyla Ascomycota, Basidiomycota and Mortierellomycota. The uncommon phyla Chytridiomycota, Rozellomycota, Monoblepharomycota, Zoopagomycota and Basidiobolomycota were detected. Unknown fungi identified at higher hierarchical taxonomic levels (Fungal sp. 1, Fungal sp. 2, Spizellomycetales sp. and Rozellomycotina sp.) and taxa identified at generic and specific levels (Mortierella sp., Pseudogymnoascus sp., Mortierella alpina, M. turficola, Neoascochyta paspali, Penicillium sp. and Betamyces sp.) dominated the assemblages. In general, the assemblages displayed high diversity and richness, and moderate dominance. Only 12 of the fungal ASVs were detected in all chronosequence soils sampled. Sequences representing saprophytic, pathogenic and symbiotic fungi were detected. Based on the sequence diversity obtained, Clearwater Mesa soils contain a complex fungal community, including the presence of fungal groups generally considered rare in Antarctica, with dominant taxa recognized as cold-adapted cosmopolitan, endemic, saprotrophic and phytopathogenic fungi. Clearwater Mesa ecosystems are impacted by the effects of regional climatic changes, and may provide a natural observatory to understand climate change effects over time.

DNA Metabarcoding Reveals Cryptic Diversity in Forest Soils on the Isolated Brazilian Trindade Island, South Atlantic.

Located 1140 km from the South American coastline in the South Atlantic Ocean and with an age of 4 million years, Trindade Island is the most recent volcanic component of Brazilian territory. Its original native vegetation has been severely damaged by human influence, in particular through the introduction of exotic grazing animals such as goats. However, since the complete eradication of goats and other feral animals in the late 1990s, the island's vegetation has been recovering, and even some endemic species that had been considered extinct have been rediscovered. In this study, we set out to characterize the contemporary cryptic diversity in soils of the recovering native forest of Trindade Island using metabarcoding by high throughput sequencing (HTS). The sequence diversity obtained was dominated by microorganisms, including three domains (Bacteria, Archaea, and Eukarya) and five kingdoms (Fungi, Metazoa, Protozoa, Chromista, and Viridiplantae). Bacteria were represented by 20 phyla and 116 taxa, with Archaea by only one taxon. Fungi were represented by seven phyla and 250 taxa, Viridiplantae by five phyla and six taxa, Protozoa by five phyla and six taxa, Metazoa by three phyla and four taxa and Chromista by two phyla and two taxa. Even after the considerable anthropogenic impacts and devastation of the island's natural forest, our sequence data reveal the presence of a rich and complex diversity of microorganisms, invertebrates, and plants and provide important baseline biodiversity information that will contribute to ecological restoration efforts on the island.

Fungal and fungal-like diversity in marine sediments from the maritime Antarctic assessed using DNA metabarcoding.

We assessed the fungal and fungal-like sequence diversity present in marine sediments obtained in the vicinity of the South Shetland Islands (Southern Ocean) using DNA metabarcoding through high-throughput sequencing (HTS). A total of 193,436 DNA reads were detected in sediment obtained from three locations: Walker Bay (Livingston Island) at 52 m depth (48,112 reads), Whalers Bay (Deception Island) at 151 m (104,704) and English Strait at 404 m (40,620). The DNA sequence reads were assigned to 133 distinct fungal amplicon sequence variants (ASVs) representing the phyla Ascomycota, Basidiomycota, Mortierellomycota, Chytridiomycota, Glomeromycota, Monoblepharomycota, Mucoromycota and Rozellomycota and the fungal-like Straminopila. Thelebolus balaustiformis, Pseudogymnoascus sp., Fungi sp. 1, Ciliophora sp., Agaricomycetes sp. and Chaetoceros sp. were the dominant assigned taxa. Thirty-eight fungal ASVs could only be assigned to higher taxonomic levels, and may represent taxa not currently included in the available databases or represent new taxa and/or new records for Antarctica. The total fungal community displayed high indices of diversity, richness and moderate to low dominance. However, diversity and taxa distribution varied across the three sampling sites. In Walker Bay, unidentified fungi were dominant in the sequence assemblage. Whalers Bay sediment was dominated by Antarctic endemic and cold-adapted taxa. Sediment from English Strait was dominated by Ciliophora sp. and Chaetoceros sp. These fungal assemblages were dominated by saprotrophic, plant and animal pathogenic and symbiotic taxa. The detection of an apparently rich and diverse fungal community in these marine sediments reinforces the need for further studies to characterize their richness, functional ecology and potential biotechnological applications.

Soil-chronosequence and Quaternary landscape evolution at the marine terraces of Harmony Point, Nelson Island, Maritime Antarctica.

This study characterized the physical, chemical, macro- and micromorphological soil properties from three successive marine terrace levels from Harmony Point (Nelson Island, Maritime Antarctica) in order to understand the pedological signatures of Quaternary coastal landscape evolution of Maritime Antarctica. Soils were sampled on the Late Holocene beach (current beach) and Mid Holocene marine terraces higher up, at 3, 8, and 12 m a.s.l. At the lower levels, the predominant soils were Gelorthents, whereas Haplogelepts dominate the higher terraces. Soil properties are mostly influenced by parent material and faunal activity, in which cryoclastic (thermal weathering) and phosphatization are the main soil-forming processes. Soils from the upper levels are more developed, deeper with reddish colors, granular structures and incipient formation B horizon. These horizonation features highlight that soils vary according with age of glacier-isostatic terrace uplift, representing a Quaternary soil chronosequence. All marine terrace levels are Ornithogenic soils, at varying degrees. However, the presence of old bird nesting sites for long periods led to formation of phosphatic horizons, stable Fe-phosphate minerals and abundant vegetation in the highest terraces of this part of Maritime Antarctica.

Soil pockets phosphatization and chemical weathering of sites affected by flying birds of Maritime Antarctica.

The majority of ornithogenic soils studied in Antarctica focus on the influence of penguins, wherever little reports evaluated the influence of flying birds on soil genesis. This study aimed to characterize the morphologic, chemic, physic, mineralogic, and micromorphologic ornithogenic soil pockets influenced by flying birds in Snow Island, Maritime Antarctica. Fifteen soil pockets were selected, described, sampled and analyzed, these sites constitute the main areas with intense long-term terrestrial biological activity in Snow Island. In order to investigate the impact of phosphatization, we compared the soil pockets with the surrounding soils and soils affected by penguins. Zone of phosphatization have a high concentration of P, K, and Ca. The XRD patterns for the clay fraction of ornithogenic soils show that phosphate minerals are the main crystalline phases (leucophosphite, minyulite, fluorapatite, and apatite). We show that even under typical periglacial conditions, sites influenced by flying birds present active chemical weathering processes. The phosphatization release exchangeable bases and accelerate mineralogical and micromorphological transformations in soils. Under the current global warming trend and expected sea-level rise, the ornithogenic environments are susceptible to accelerated erosion rates and a great part of these hotspots may be lost for the open sea.

Acid sulfate soils from Antarctica: genesis and properties along a climatic gradient.

Sulfurization is a pedogenic process that involves pyrite oxidation and strong soil acidification, accounting for the formation of acid sulfate soils. In Antarctica, acid sulfate soils are related to specific parent materials, such as sulfide-bearing andesites in Maritime Antarctica and pyritized sedimentary rocks in James Ross Archipelago. The hypothesis is that the acid sulfate soils of these regions vary according with a climate gradient. The reviewing of current data showed that the acid sulfate soils of warmer and wetter Maritime Antarctica have a greater weathering degree, higher acidity, leaching, phosphorus adsorption capacity, structural development, and well-crystallized iron oxides and kaolinite formation. On the other hand, the sulfurization at the drier region of James Ross Archipelago is counterbalanced by the semiaridity, resulting in lower acidity and higher base contents combined with little morphological and mineralogical evolution besides presence of weatherable minerals in the clay fraction. The sulfurization process interplays with other pedogenic processes, such as the phosphatization in Maritime Antarctica and salinization in James Ross Archipelago. Higher temperatures and soil moisture enhance the pedogenesis, showing that even the Antarctic sulfate soils, which originated from specific parent materials, have their development and characteristics controlled by a clear climatic gradient.

Diversity and ecology of cultivable fungi isolated from the thermal soil gradients in Deception Island, Antarctica.

We surveyed the diversity of cultivable fungi isolated from cold and hot volcanic soils of Deception Island, Antarctica. Seventy-four fungal isolates were identified; these belonged to 17 taxa in the genera Aspergillus, Penicillium, Pseudogymnoascus, Purpureocillium, and Mortierella. The fungal assemblages showed low diversity, richness, and dominance indices. The Aspergillus taxa were dominant in the soils at 0 °C, 50 °C, and 100 °C. Aspergillus lacticoffeatus, Aspergillus cf. ruber, Penicillium citrinun, and Purpureocillium sodanum were present only in soils having a temperature of 100 °C. Aspergillus calidoustus was present in all thermal soils and displayed the highest densities. The majority of fungi displayed mesophilic behavior; however, different isolates of Aspergillus lacticoffeatus and Aspergillus niger were able to grow at 50 °C; these are phylogenetically close to the causative agents of aspergillosis in immunocompromised individuals. Deception Island perhaps represents one of the most visited regions in Antarctica and the tourism there has increased over the last 20 years, especially by elderly tourists, probably with weak immune systems, come in contact with the resident microorganisms, including the thermo-resistant opportunistic Aspergillus species.

The diversity, distribution, and pathogenic potential of cultivable fungi present in rocks from the South Shetlands archipelago, Maritime Antarctica.

We studied the molecular taxonomy and diversity of cultivable rock fungi from Antarctic islands. From 50 rock samples, 386 fungal isolates were obtained and identified as 33 taxa of 20 genera. The genera Cladophialophora, Cladosporium, Cyphellophora, Eichleriella, Paracladophialophora, and Penicillium displayed the highest densities. Ecological diversity indices showed that the fungal assemblages are diverse and rich with low dominance. The genera Cladophialophora, Cladosporium, and Penicillium showed a broad distribution from rocks of the various islands. One hundred and fifty-nine fungi, grown at 37 °C, were identified as Penicillium chrysogenum, Fusarium sp., and Rhodotorula mucilaginosa. One hundred and three fungi displayed haemolytic activity, 81 produced proteinase, 9 produced phospholipase, and 25 presented dimorphism and a spore diameter ≤ 4 µm. The Antarctic Peninsula region appears to be under the effects of global climate changes, which may expose and accelerate the rock's weathering processes, and expose and release cryptic fungi and other microbes, especially those with innate pathogenic potential, previously arrested in rocks. Consequently, these rocks and their particles may represent a vehicle for the dispersal of microbial propagules, including those able to spread pathogens, along, across, and out of Antarctica.

Cultivable fungi present in Antarctic soils: taxonomy, phylogeny, diversity, and bioprospecting of antiparasitic and herbicidal metabolites.

Molecular biology techniques were used to identify 218 fungi from soil samples collected from four islands of Antarctica. These consisted of 22 taxa of 15 different genera belonging to the Zygomycota, Ascomycota, and Basidiomycota. Mortierella, Antarctomyces, Pseudogymnoascus, and Penicillium were the most frequently isolated genera and Penicillium tardochrysogenum, Penicillium verrucosus, Goffeauzyma gilvescens, and Mortierella sp. 2 the most abundant taxa. All fungal isolates were cultivated using solid-state fermentation to obtain their crude extracts. Pseudogymnoascus destructans, Mortierella parvispora, and Penicillium chrysogenum displayed antiparasitic activities, whilst extracts of P. destructans, Mortierella amoeboidea, Mortierella sp. 3, and P. tardochrysogenum showed herbicidal activities. Reported as pathogenic for bats, different isolates of P. destructans exhibited trypanocidal activities and herbicidal activity, and may be a source of bioactive molecules to be considered for chemotherapy against neglected tropical diseases. The abundant presence of P. destructans in soils of the four islands gives evidence supporting that soils in the Antarctic Peninsula constitute a natural source of strains of this genus, including some P. destructans strains that are phylogenetically close to those that infect bats in North America and Europe/Palearctic Asia.

Influence of growth regulators on distribution of trichomes and the production of volatiles in micropropagated plants of Plectranthus ornatus

ABSTRACT The profile of volatile organic compounds, the glandular and non-glandular trichomes of Plectranthus ornatus, obtained by in vitro cultivation, was evaluated in plants grown in Murashide and Skoog medium supplemented with benylaminopurine at 4.5, 9.0, and 18.0 µM + naphthaleneacetic acid at 5.37 µM, kinetin at 4.7, 9.3 and 18.5 µM + naphthaleneacetic acid (5.37 µM) or Murashide and Skoog 0 medium (as a control). Scanning Electron Microscopy was performed on samples of the third leaf node of the 90 days old plants obtained from treatment with 4.5 or 9.0 µM benylaminopurine, and 4.7 or 9.3 µM kinetin. Headspace Solid Phase Micro-Extraction of the 30, 60 and 90 days old in vitro plants permitted to determinate by GC/MS the composition comprised of 62 compounds. The data were analyzed using Principal Component Analysis and Hierarchical Clustering Analysis and, the major constituents of these oils after treatment and aging were monoterpenes and sesquiterpenes. Morphoanatomical analysis of trichomes, by Scanning Electron Microscopy, enabled the identification of non-glandular trichomes and four types of glandular trichomes, which comprised capitate and peltate glandular trichomes that were distributed on both sides of the leaf. We observed that the regulators influenced qualitative and quantitative profiles of the volatile organic compounds and the number and distribution of hairs on the leaf surface.

Antarctic rocks from continental Antarctica as source of potential human opportunistic fungi.

We assessed the diversity of culturable fungi associated with rocks of continental Antarctica to evaluate their physiological opportunistic virulence potential in vitro. The seventy fungal isolates obtained were identified as nine species of Acremonium, Byssochlamys, Cladosporium, Debaryomyces, Penicillium, and Rhodotorula. Acremonium sp., D. hansenii, P. chrysogenum, P. citrinum, P. tardochrysogenum, and R. mucilaginosa were able to grow at 37 °C; in addition, B. spectabilis displayed a high level of growth at 37 and 45 °C. Thirty-one isolates of P. chrysogenum, P. citrinum, and P. tardochrysogenum were able to produce partial haemolysis on blood agar at 37 °C. Acremonium sp., P. citrinum, and P. tardochrysogenum showed spore sizes ranging from 2.81 to 5.13 µm diameters at 37 °C. Of these, P. chrysogenum and P. tardochrysogenum displayed macro- and micro morphological polymorphism. Our results suggest that rocks of the ultra-extreme cold and dry environment of Antarctica harbour cryptic fungi phylogenetically close to opportunistic pathogenic and mycotoxigenic taxa with physiologic virulence characteristics in vitro.

Fungi associated with rocks of the Atacama Desert: taxonomy, distribution, diversity, ecology and bioprospection for bioactive compounds.

This study assessed the diversity of cultivable rock-associated fungi from Atacama Desert. A total of 81 fungal isolates obtained were identified as 29 Ascomycota taxa by sequencing different regions of DNA. Cladosporium halotolerans, Penicillium chrysogenum and Penicillium cf. citrinum were the most frequent species, which occur at least in four different altitudes. The diversity and similarity indices ranged in the fungal communities across the latitudinal gradient. The Fisher-α index displayed the higher values for the fungal communities obtained from the siltstone and fine matrix of pyroclastic rocks with finer grain size, which are more degraded. A total of 23 fungal extracts displayed activity against the different targets screened. The extract of P. chrysogenum afforded the compounds α-linolenic acid and ergosterol endoperoxide, which were active against Cryptococcus neoformans and methicillin-resistance Staphylococcus aureus respectively. Our study represents the first report of a new habitat of fungi associated with rocks of the Atacama Desert and indicated the presence of interesting fungal community, including species related with saprobes, parasite/pathogen and mycotoxigenic taxa. The geological characteristics of the rocks, associated with the presence of rich resident/resilient fungal communities suggests that the rocks may provide a favourable microenvironment fungal colonization, survival and dispersal in extreme conditions.

Volatile organic compounds obtained by in vitro callus cultivation of Plectranthus ornatus Codd. (Lamiaceae).

Plectranthus spp (Lamiaceae) are plants of economic importance because they are sources of aromatic essential oils and are also cultivated and several species of this genus are used as folk medicines. This paper describes the effects of different concentrations of the 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthaleneacetic acid (NAA) on the induction of callus from nodal segments of Plectranthus ornatus Codd and in the production of volatile organic compounds (monoterpenes and sesquiterpenes). The 20 and 40 day calli were subjected to solid phase micro extraction (HS-SPME) and submitted to GCMS analysis. Variations in VOCs between the samples were observed and, a direct relationship was observed between of the major constituent detected (α-terpinyl acetate) and the monoterpenes α-thujene, α-pinene, ß-pinene, camphene, sabinene and α-limonene that were present in the volatile fractions. Besides α-terpinyl acetate, isobornyl acetate and α-limonene were also major constituents. Variations were observed in VOCs in the analyzed periods. The best cultivation media for the production of VOCs was found to be MS0 (control). Moderate success was achieved by treatment with 2.68 µM and 5:37 µM NAA (Group 2). With 2,4-D (9.0 µM), only the presence of α-terpinyl acetate and isocumene were detected and, with 2.26 µM of 2,4-D was produced mainly α-terpinyl acetate, α-thujene and ß-caryophyllene (16.2%). The VOC profiles present in P. ornatus were interpreted using PCA and HCA. The results permitted us to determine the best cultivation media for VOC production and, the PCA and HCA analysis allowed us to recognize four groups among the different treatments from the compounds identified in this set of treatments.

Headspace solid phase microextraction/gas chromatography-mass spectrometry combined to chemometric analysis for volatile organic compounds determination in canine hair: a new tool to detect dog contamination by visceral leishmaniasis.

A new analytical methodology using HS-SPME/GC-MS was optimized in order to attain maximum sensitivity, using multivariate strategies. The proposed method was employed to evaluate the VOC profile exhaled from canine hair samples collected from 8 healthy dogs and from 16 dogs infected by Leishmania infantum. 274 VOCs were detected, which could be identified as aldehydes, ketones and hydrocarbons. After application of the Soft Independent Modeling of Class Analogy (SIMCA) and Principal Component Analysis (PCA) healthy and infected dogs, with similar VOCs profiles, could be separately grouped, based on compounds such as 2-hexanone, benzaldehyde, and 2,4-nonadienal. The proposed method is non-invasive, painless, readily accepted by dog owners and could be useful to identify several biomarkers with applications in the diagnosis of diseases.

Multivariate optimization of a GC-MS method for determination of sixteen priority polycyclic aromatic hydrocarbons in environmental samples.

This paper describes the development and optimization, by using multivariate analysis, of a GC-MS-SIM method for evaluation of the 16 polyaromatic hydrocarbons considered as priority pollutants in atmospheric particulate material by the US EPA. In order to assure an adequate separation in the shortest analysis time, a multivariate design was used to set the conditions of the oven temperature program. The optimization process was carried out using factorial fractional design and Box-Behnken design. The following factors were evaluated: initial temperature, temperature rate #1, intermediary temperature, temperature rate #2, and final temperature. The optimized conditions were set at: 70 degrees C (2 min) --> 200 degrees C (30 degrees C/min, 5 min) --> 300 degrees C (5 degrees C/min, 1.67 min). Moreover, we have also optimized the injector temperature as 310 degrees C and sampling time as 0.8 min. The total analysis time was 33 min. Validation of GC-MS-SIM yielded satisfactory results for repetitivity of the detector response and retention times, and linearity of calibration curves. LOD were established as 0.13-0.34 ng/mL (peak area) and 0.18-0.72 ng/mL (peak height). The method has been shown to be appropriate for the analysis of samples of atmospheric particulate material and/or other environmental matrices.