Near‐shore microbial communities (Eukaryotes, Bacteria and Archaea) of the sub‐Antarctic Prince Edward Islands

最新版本 由 SCAR - Microbial Antarctic Resource System 發佈於 Mar 19, 2019 SCAR - Microbial Antarctic Resource System

Amplicon sequencing dataset of microbial eukaryotes (18S ssh rRNA); bacteria (16S) and Archaea (16S) from coastal seawater near the shore of the Prince Edward Islands (Indian Ocean, Sub-Antarctica); sampled from a single location (37.58 degrees South 46.36 degrees East) in 2012, 2013, 2014 and 2015.

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版本

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如何引用

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Venkatachalam S, Matcher G, Lamont T, Dorrington R (2018): Near‐shore microbial communities (Eukaryotes, Bacteria and Archaea) of the sub‐Antarctic Prince Edward Islands. v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=marine_microbial_communities_princeeedward_islands&v=1.2

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The publisher and rights holder of this work is 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: 576f1f95-2ad4-4d23-a052-c02d901929f1。  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

資源建立者:

Siddarthan Venkatachalam
Rhodes University Grahamstown ZA
Gwynneth Matcher
Rhodes University Grahamstown ZA
Tarron Lamont
Department of Environmental Affairs Cape Town ZA
Rosemary Dorrington
Rhodes University Grahamstown ZA

可回覆此資源相關問題者:

Rosemary Dorrington
Rhodes University Grahamstown ZA

元數據填寫者:

Maxime Sweetlove
Research assistent
Royal Belgian Institute for Natural Sciences Rue Vautier 29 1000 Brussels BE

與此資源的相關者:

使用者

地理涵蓋範圍

Sampled near the Prince Edward Islands (Indian Ocean, Sub-Antarctica); 37.58 degrees South 46.36 degrees East

界定座標範圍 緯度南界 經度西界 [-37.58, 46.36], 緯度北界 經度東界 [-37.58, 46.36]

分類群涵蓋範圍

Microbial Eukaryotes (18S ssh rRNA gene, v9), Bacteria (16S ssh rRNA gene, v4-v5) and Archaea (16S ssh rRNA gene, v4-v5)

Domain  Eukaryota (Eukaryotes),  Bacteria (Bacteria),  Archaea (Archaea)

時間涵蓋範圍

彙整期間 2012-2015

計畫資料

無相關描述

計畫名稱 Influence of oceanographic variability on near‐shore microbial communities
經費來源 This project was funded by grants from the South African National Antarctic Programme (SANAP) through the South African National Research Foundation (NRF) to R.A.D. (80260) and I.J.A. (80270) and the Rhodes University Sandisa Imbewu Programme; the DST/NRF SARChI Post‐Doctoral Fellowship (87583); the South African Department of Environmental Affairs (DEA) and the University of Cape Town and Rhodes University.

The personnel involved in the project:

Siddarthan Venkatachalam

取樣方法

Two liters of surface (5 m depth) seawater was initially filtered through 100 μm mesh to remove particulate matter, after which microbial biomass was collected by filtration through 0.22 μm Polyethersulfone (PES) membrane (Pall Corporation). The filters were immersed in RNALater (Qiagen) and stored at −20°C.

研究範圍 Seawater samples were collected at a near‐shore site on the northeast coast of PEI at 46°36.415′S; 37°58.553′E during the austral autumn (April) and winter (July) seasons in 2012 as well as the austral autumn (May) for years 2013, 2014, and 2015.

方法步驟描述:

  1. Genomic DNA (gDNA) and RNA were extracted from PES filters using the AllPrep DNA/RNA Mini Kit (Qiagen) according to the manufacturer's instructions. For the rRNA sequencing, isolated RNA was converted to cDNA using the QuantiTect Reverse Transcription Kit (Qiagen) according to the manufacturer's protocol. For the analysis of bacterial community composition, the V4‐V5 variable regions of the bacterial 16S rRNA gene were amplified by PCR using the E517F (5′‐CAG CAG CCG CGG TAA‐3′) and E969‐984R (5′‐GTA AGG TTC YTC GCG T‐3′) primers) with suitable multiplex identifier tags and sequencing primer binding sites attache. For the analysis of eukaryotic phytoplankton community diversity, PCR amplification of 18S rRNA gene sequences was carried out using primers 1391F: 5′‐GTA CAC ACC GCC CGT C‐3′ (Saccharomyces cerevisiae position 1629–1644) and EukB: 5′‐TGA TCC TTC TGC AGG TTC ACC TAC‐3′ (S. cerevisiae position 1774–1797) targeting the V9 regions of the eukaryotic SSU rRNA. Archaeal 16S rRNA gene sequences (V4‐V5 variable regions) were amplified using pr514–528: 5′‐GGT GYC AGC CGC CGC‐3′ and A906R: 5’‐CCC GCC AAT TCC TTT AAG TTTC‐3, respectively.
  2. PCR amplification of the bacteria, phytoplankton and archaeal gene fragments was carried out in a 25 μL reaction volume comprising 10 ng of the extracted DNA and using KAPAHiFi Hotstart DNA Polymerase (KAPA Biosystems) according to the manufacturer's instructions. For bacterial 16S rRNA gene amplification, the reaction mixtures were subjected to the reaction conditions described in Matcher et al. (2011). For 18S rRNA amplification, the PCR cycling parameters were as follows: 98°C (45 s), 57°C (30 s), 72°C (45 s) for five cycles, 98°C (45 s), 65°C (30 s), and 72°C (45 s) for 15 cycles and a final extension at 72°C for 5 min. For archaeal 16S rRNA gene amplification, cycling conditions were used as for the 18S rRNA amplification with amendments of the annealing temperatures to 56°C (30 s) for the first five cycles and 59°C (30 s) for next 15 cycles.
  3. The PCR amplification products were gel‐purified using the ISOLATE II PCR and Gel Kit (Bioline), subjected to emulsion PCR, and then sequenced using the GS Junior Titanium Sequencer (454 Life Sciences, Roche).

引用文獻

  1. Venkatachalam, S., Matcher, G. F., Lamont, T., van den Berg, M., Ansorge, I. J., & Dorrington, R. A. (2018). Influence of oceanographic variability on near‐shore microbial communities of the sub‐Antarctic Prince Edward Islands. Limnology and Oceanography.

額外的元數據

替代的識別碼 576f1f95-2ad4-4d23-a052-c02d901929f1
https://ipt.biodiversity.aq/resource?r=marine_microbial_communities_princeeedward_islands