fumarolic microbial communities at Tramway Ridge, Mt. Erebus, Antarctica

Última versión publicado por SCAR - Microbial Antarctic Resource System el mar 19, 2019 SCAR - Microbial Antarctic Resource System
Fecha de publicación:
19 de marzo de 2019
Licencia:
CC-BY 4.0

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Descripción

Microbial (Bacteria) Dataset containing amplicon sequencing samples and metagenome shotgun sequencing samples from two fumarole soil location at Tramway Ridge, Mount Erebus, Antarctica (ASPA 130)

Versiones

La siguiente tabla muestra sólo las versiones publicadas del recurso que son de acceso público.

¿Cómo referenciar?

Los usuarios deben citar este trabajo de la siguiente manera:

Herbold C, Lee C, McDonald I, Cary C (2019): fumarolic microbial communities at Tramway Ridge, Mt. Erebus, Antarctica. v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=fumarolic_antarctic_microbial_communities_at_mount_erebus&v=1.2

Derechos

Los usuarios deben respetar los siguientes derechos de uso:

El publicador y propietario de los derechos de este trabajo es SCAR - Microbial Antarctic Resource System. Esta obra está bajo una licencia Creative Commons de Atribución/Reconocimiento (CC-BY 4.0).

Registro GBIF

Este recurso ha sido registrado en GBIF con el siguiente UUID: 53b30953-201f-4599-9377-1c41b5cba477.  SCAR - Microbial Antarctic Resource System publica este recurso y está registrado en GBIF como un publicador de datos avalado por Scientific Committee on Antarctic Research.

Palabras clave

Metadata

Contactos

Craig Herbold
  • Originador
  • Punto De Contacto
The University of Waikato
Hamilton
NZ
Charles Lee
  • Originador
The University of Waikato
Hamilton
NZ
Ian McDonald
  • Originador
The University of Waikato
Hamilton
NZ
Craig Cary
  • Originador
The University of Waikato
Hamilton
NZ
Maxime Sweetlove
  • Proveedor De Los Metadatos
Research assistent
Royal Belgian Institite for Natural Sciences
Rue Vautier 29
1000 Brussels

Cobertura geográfica

Tramway Ridge (ASPA 130), Mount Erebus, Antarctica

Coordenadas límite Latitud Mínima Longitud Mínima [-77,518, 167,111], Latitud Máxima Longitud Máxima [-77,518, 167,111]

Cobertura taxonómica

metagenomic data (amplicon and shotgun) of Bacteria

Dominio Bacteria (Bacteria)

Datos del proyecto

No hay descripción disponible

Título Evidence of global-scale aeolian dispersal and endemism in isolated geothermal microbial communities of Antarctica
Fuentes de Financiación Financial support was provided by grant UOW0802 from the New Zealand Marsden Fund. Additional salary support was provided by the New Zealand Marsden fund to C.K.L. (UOW1003) and C.W.H. (UOW0802).

Personas asociadas al proyecto:

Craig Herbold

Métodos de muestreo

All suggested sterilization protocols for entering into this protected site were adhered to, following the ASPA 130 Management Plan ( http://www.scar.org/publications/bulletins/151/aspa130.html). Sites were chosen based on measuring a surface temperature of 65 °C with a stainless steel Checktemp1 temperature probe (Hanna Instruments, Rhode Island, USA), sterilized with 70% ethanol immediately before each use. Surface ‘crust’ was set aside before collecting samples. Samples were collected by aseptically removing the top 2 cm of sediment in an ~25 cm2 area. Sediment was placed into a fresh 50 ml Falcon tube. Sampling continued with the collection of a second (2–4 cm depth) and third (4–8 cm depth) layer following the same procedures. Temperature measurements were repeated for each layer sampled. All samples were immediately frozen, transported back to the University of Waikato frozen and maintained at −80 °C in the laboratory until analysed.

Área de Estudio Sediment samples were collected within the Tramway Ridge Antarctic Specially Protected Area (ASPA 130) in February 2009 from two sites (site A (active site): 77° 31.103′ S, 167° 6.682′ S and site B (passive site): 77° 31.106′ S, 167° 6.668′ E).

Descripción de la metodología paso a paso:

  1. DNA was extracted from samples using a modified CTAB (cetyltrimethylammonium bromide) bead-beating protocol. For shotgun sequencing, a portion of extracted genomic DNA was sequenced using standard protocols for the 454-Ti platform (Roche 454 Life Sciences, Branford, CT, USA) at the UCLA GenoSeq CORE.
  2. PCR amplicons containing V5–V7 hypervariable regions of the 16S rRNA gene were generated from the same genomic DNA samples using primers Tx9 (5′-GGATTA GAWACCCBGGTAGTC-3′) and 1391R (5′-GACGGGCRGTGWGTRCA-3′). PCR was performed in triplicate on each sample and pooled to reduce stochastic variation. Three samples (site A 0–2 cm, site B 0–2 cm, site B 2–4 cm) were sequenced using the 454-GS-FLX platform by Taxon Biosciences (Tiburon, CA, USA) and three samples (site A 2–4 cm, site A 4–8 cm and site B 4–8 cm) were sequenced using the 454 Junior platform at the Waikato DNA Sequencing Facility (Hamilton, New Zealand). For the three samples sequenced using the 454-GS-FLX platform, each 30 μl reaction contained 2–10 ng of DNA extract, Pfx polymerase and platinum polymerase (0.5 U each; Invitrogen), 1 × Pfx PCR buffer with Pfx enhancer, 0.2 mM dNTPs, 1 mM MgCl2, 0.02 mg ml−1 BSA, 0.8 μM of forward and reverse primer and PCR-grade water. Thermal cycling conditions were 94 °C for 2 min; 24 cycles of 94 °C for 15 s, 55 °C for 30 s and 68 °C for 1 min; and 68 °C for 3 min. Amplicons were size-selected and purified using polyacrylamide gel electrophoresis before being prepared for pyrosequencing by Taxon Biosciences. For the three samples sequenced using the 454-junior platform, each 30 μl reaction contained 2–10 ng of DNA extract, PrimeStar polymerase (0.625 U; Takara), 1 × PCR buffer, 0.2 mM dNTPs, 0.4 μM of forward and reverse primer and PCR-grade water. Thermal cycling conditions were 94 °C for 3 min; 24 cycles of 94 °C for 20 s, 52 °C for 20 s and 72 °C for 45 s; and 72 °C for 3 min. Triplicate PCR reactions were pooled and gel-purified using the UltraCleanTM 15 DNA Purification Kit (MO BIO Laboratories Inc.), cleaned using the Agencourt AMPure XP Bead Cleanup kit (Beckman Coulter Inc.) and quantified (Quant-iTTM dsDNA HS Assay Kit, Invitrogen Ltd.). Cleaned amplicons were used as template (25 ng) in a second PCR reaction using fusion primers (forward: 5′-(454 Adapter A)-TCAG-MID-Tx9-3′; reverse: 5′-(454 Adapter B)-TCAG-1391R-3′). PCR conditions were exactly the same as the first round except only 10 cycles of PCR were performed. Triplicate PCR reactions were pooled and gel-purified using the UltraCleanTM 15 DNA Purification Kit, cleaned using the Agencourt AMPure XP Bead Cleanup kit and quantified (Quant-iTTM dsDNA HS Assay Kit) before being prepared for pyrosequencing using the 454 Junior platform by the University of Waikato sequencing facility.

Referencias bibliográficas

  1. Herbold, C. W., Lee, C. K., McDonald, I. R., & Cary, S. C. (2014). Evidence of global-scale aeolian dispersal and endemism in isolated geothermal microbial communities of Antarctica. Nature communications, 5, 3875.

Metadatos adicionales

Identificadores alternativos 53b30953-201f-4599-9377-1c41b5cba477
https://ipt.biodiversity.aq/resource?r=fumarolic_antarctic_microbial_communities_at_mount_erebus