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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此資料的發布者及權利單位為 SCAR - Microbial Antarctic Resource System。 This work is licensed under a Creative Commons Attribution (CC-BY 4.0) License.

GBIF 註冊

此資源已向GBIF註冊，並指定以下之GBIF UUID: 11c8e968-361d-4cea-abae-67450fa03fe8。  SCAR - Microbial Antarctic Resource System 發佈此資源，並經由Scientific Committee on Antarctic Research同意向GBIF註冊成為資料發佈者。

關鍵字

Metadata

聯絡資訊

地理涵蓋範圍

South Shetland Islands, Union Glacier (Antarctica)

分類群涵蓋範圍

LSU marker gene for microbial soil Fungi

時間涵蓋範圍

計畫資料

無相關描述

參與計畫的人員:

取樣方法

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.

方法步驟描述:

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.

引用文獻

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.

額外的詮釋資料