Fungi (LSU) in soils from the South Shetland island, Antarctic Peninsula island, and Union Glacier

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Baeza M, Barahona S, Alcaino J, Cifuentes V (2018): Fungi (LSU) in soils from the South Shetland island, Antarctic Peninsula island, and Union Glacier. v1.3. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=fungi_soils_s_shetland_and_antarctica&v=1.3

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L’éditeur et détenteur des droits de cette ressource est SCAR - Microbial Antarctic Resource System. Ce travail est sous licence Creative Commons Attribution (CC-BY) 4.0.

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Cette ressource a été enregistrée sur le portail GBIF, et possède l'UUID GBIF suivante : 11c8e968-361d-4cea-abae-67450fa03fe8.  SCAR - Microbial Antarctic Resource System publie cette ressource, et est enregistré dans le GBIF comme éditeur de données avec l'approbation du Scientific Committee on Antarctic Research.

Mots-clé

Metadata

Contacts

Couverture géographique

South Shetland Islands, Union Glacier (Antarctica)

Couverture taxonomique

LSU marker gene for microbial soil Fungi

Couverture temporelle

Données sur le projet

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Les personnes impliquées dans le projet:

Méthodes d'échantillonnage

Samples were collected in sterile 50 ml plastic tubes, which were sealed and shipped at -20°C to the Genetics Laboratory of the Faculty of Science at the Universidad de Chile. Once samples arrived at the laboratory, they were maintained at -80°C until processing.

Description des étapes de la méthode:

1. A PowerSoil DNA Isolation Kit (MO BIO Laboratories Inc., Carlsbad, CA, United States) was used for direct DNA extraction from soil samples. The manufacturers’ instructions were followed, with only one modification, the disruption step, was performed using a Mini Beadbeater-16 cell disrupter (BioSpec Bartlesville, United States) instead of vortex agitation, as no PCR-amplicons were obtained in reactions using samples obtained through vortex agitation.
2. The PCR reactions were performed using 1 μl of DNA sample (direct or 1/10 dilution), 5 units of Taq DNA polymerase, dNTP mix at 0.4 mM each, forward and reverse primers at 1 mM final each, PCR buffer and MgCl2 2 mM. Amplification was performed using a 2720 (Applied Biosystems) thermal cycler using the following protocol: initial denaturation at 94°C for 3 min; 35 cycles of denaturation at 94°C for 30 s, annealing at 50°C for 3 min, and extension at 72°C for 3 min; and a final extension step at 72°C for 10 min. The universal primers F63 (5′-GCA TAT CAA TAA GCG GAG GAA AAG-3′) and LR3 (5′-GGT CCG TGT TTC AAG ACG G-3′) were used; these primers have been described as specific for amplifying the fungal D1/D2 region of the large subunit ribosomal gene (LSU). In all PCR reactions, the same forward primer (F63) was used, but the reverse primer (LR3) was specific for each soil sample as a 454 adapter, a specific barcode and a linker sequence were added.
3. Sequencing was performed at OMICS Solutions (Santiago, Chile) using the Ion 314TM Chip Kit v2 (Thermo Fisher) and Ion Torrent personal genome machine (PGM), according to the manufacturer’s instructions (Rothberg et al., 2011). Three independent runs were performed: (i) samples from King George Island; (ii) samples from Deception, Snow, Dee, Livingstone, Greenwich, Robert, Nelson and Litchfield islands; and iii) samples from Lagotellerie and Union Glacier.

Citations bibliographiques

1. Baeza, M., Barahona, S., Alcaíno, J., & Cifuentes, V. (2017). Amplicon-Metagenomic Analysis of Fungi from Antarctic Terrestrial Habitats. Frontiers in microbiology, 8, 2235.
2. Carrasco, M., Rozas, J. M., Barahona, S., Alcaíno, J., Cifuentes, V., & Baeza, M. (2012). Diversity and extracellular enzymatic activities of yeasts isolated from King George Island, the sub-Antarctic region. BMC microbiology, 12(1), 251.
3. Troncoso, E., Barahona, S., Carrasco, M., Villarreal, P., Alcaíno, J., Cifuentes, V., & Baeza, M. (2017). Identification and characterization of yeasts isolated from the South Shetland Islands and the Antarctic Peninsula. Polar Biology, 40(3), 649-658.
4. Barahona, S., Yuivar, Y., Socias, G., Alcaíno, J., Cifuentes, V., & Baeza, M. (2016). Identification and characterization of yeasts isolated from sedimentary rocks of Union Glacier at the Antarctica. Extremophiles, 20(4), 479-491.

Métadonnées additionnelles