Cyanobacteria in microbial mats from Antarctic lakes

Dernière version Publié par SCAR - Microbial Antarctic Resource System le Mar 19, 2019 SCAR - Microbial Antarctic Resource System

Amplicon sequencing dataset targeting Cyanobacteria (16S ssu rRNA gene) in microbial benthic mats from 13 lakes across the Antarctic continent.

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Pessi I, Lara Y, Durieu B, de C. Maarouf P, Verleyen E, Wilmotte A (2019): Cyanobacteria in microbial mats from Antarctic lakes. v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=lacustrine_cyanobacteria_antarctica&v=1.2

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L’éditeur et détenteur des droits de cette ressource est SCAR - Microbial Antarctic Resource System. This work is licensed under a Creative Commons Attribution (CC-BY) 4.0 License.

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Cette ressource a été enregistrée sur le portail GBIF, et possède l'UUID GBIF suivante : a41ad9bd-cf79-48fa-89a0-102ddf4d4e61.  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

Personne ayant créé cette ressource:

Igor Pessi
University of Liège Alle ́e du Six Aouˆt 13, B6a 4000 Liège BE
Yannick Lara
University of Liège Alle ́e du Six Aouˆt 13, B6a 4000 Liège BE
Benoit Durieu
University of Liège Alle ́e du Six Aouˆt 13, B6a 4000 Liège BE
Pedro de C. Maarouf
University of Liège Alle ́e du Six Aouˆt 13, B6a 4000 Liège BE
Elie Verleyen
Ghent University Krijgslaan 281 9000 Ghent BE
Annick Wilmotte
University of Liège Alle ́e du Six Aouˆt 13, B6a 4000 Liège BE

Personne pouvant répondre aux questions sur la ressource:

Igor Pessi
University of Liège Alle ́e du Six Aouˆt 13, B6a 4000 Liège BE

Personne ayant renseigné les métadonnées:

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

Autres personnes associées à la ressource:

Utilisateur

Couverture géographique

Circum-Antarctic sampling of benthic microbial mats in lakes (n=13)

Enveloppe géographique Sud Ouest [-82.45, -67.233], Nord Est [-66.283, 100.233]

Couverture taxonomique

Cyanobacteria (16S ssu rRNA gene)

Phylum  Cyanobacteria (Cyanobacteria)

Couverture temporelle

Epoque de formation 1997-2007

Données sur le projet

CCAMBIO (Climate Change and Antarctic Microbial Biodiversity) is an academic project funded by the Belgian Federal Science Policy (BELSPO). Its main objective is to study the diversity, biogeographic zoning and genomic make-up of lacustrine microbial mat communities in the Antarctic Realm. CCAMBIO is composed by researchers from four Belgian Universities and Institutes (University of Liège, Ghent University, National Botanical Garden of Belgium and Royal Belgian Institute of Natural Sciences), as well as collaborators from the British Antarctic Survey.

Titre Climate Change and Antarctic Microbial Biodiversity
Identifiant CCAMBIO
Financement Belgian Science Policy Office (BelSPO) project SD/BA/03
Description du domaine d'étude / de recherche Microbial mats in the benthic and littoral zon of lakes in polar environments.
Description du design Lakes from different regions in Antarctica were sampled, and sequencing was preformed of the 16S marker genes to provide a base-line inventory of microbial eukaryotes biodiversity and test hypotheses about biogeography and species distributions

Les personnes impliquées dans le projet:

Igor Pessi

Méthodes d'échantillonnage

Microbial mat samples were collected in the littoral or deeper parts of the euphotic zone of the lakes using a spatula or gravity corer, respectively. The upper 1 cm of the core was aseptically removed and kept dark and cool until transfer to -20°C.

Etendue de l'étude Samples were taken from benthic microbial mats (upper 1 cm), collected in 13 lakes on the Antarctic continent, distributed across eight Antarctic regions belonging to four distinct Antarctic Conservation Biogeographical Regions (ACBR).
Contrôle qualité DNA concentration and quality were determined using a NanoVue spectrophotometer (GE Healthcare Life Sciences, Little Chalfont, UK). A blank DNA extraction consisting of sterile Milli-Q water was carried out in parallel.

Description des étapes de la méthode:

  1. DNA was extracted from the mats using the PowerSoil DNA Isolation Kit (MOBIO Labora- tories, Carlsbad, CA, USA) according to the manufacturer’s instructions with some modifications. Tubes were agitated on a vortex for 20 extra min to ensure a good disintegration of the mats and, if not completely disintegrated, a sterile pestle was used to crush the remaining pieces.
  2. The cyanobacteria-specific primer set CYA359F and CYA781R(a)/CYA781R(b) was used to amplify the V3-V4 variable region of the 16S rRNA gene. PCR reactions consisted of 19 PCR buffer with 1.5 mM MgCl2, 1mg mL 1BSA, 200 uM of each dNTP, 0.2 uM of each primer, 1 U SUPER TAQ plus DNA polymerase (HT Biotechnology, Cambridge, UK), and 4 ng uL^-1 template DNA in a final volume of 50 uL. Amplification was performed using an initial denaturation step at 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 60°C (for primer 781Ra) or 57°C (for primer 781Rb) for 45 s and 68°C for 45 s, and a final elonga- tion at 68°C for 5 min. Negative controls (PCR mixes with either no DNA or the blank DNA extracts) were always included during PCR amplifications. To minimize stochastic PCR bias, amplification was carried out as six independent PCR reactions (three for each reverse primer) that were pooled before purification.
  3. Replicate PCR reactions were pooled and purified using the GeneJet PCR Purification Kit (Thermo Scientific, Wal- tham, MA, USA). Purified amplicons were quantified using the Quant-iT PicoGreen dsDNA Assay Kit, pooled in equimolar concentrations, and concentrated to 25 uL using the Ami- con Ultra-0.5 mL 30K device (EMD Millipore, Billerica, MA, USA). Pooled libraries were sent to Beckman Coulter Geno- mics (Takeley, UK), where primer dimers were removed using the Agencourt AMPure XP Kit (Beckman Coulter, Brea, CA, USA) and sequencing adapters were ligated to the ampli- cons. Sequences were obtained using the 454 GS FLX+ Titanium platform (454 Life Sciences, Branford, CT, USA).

Citations bibliographiques

  1. Pessi, I. S., Maalouf, P. D. C., Laughinghouse IV, H. D., Baurain, D., & Wilmotte, A. (2016). On the use of high‐throughput sequencing for the study of cyanobacterial diversity in Antarctic aquatic mats. Journal of phycology, 52(3), 356-368.
  2. Pessi, I. S., Lara, Y., Durieu, B., Maalouf, P. D. C., Verleyen, E., & Wilmotte, A. (2018). Community structure and distribution of benthic cyanobacteria in Antarctic lacustrine microbial mats. FEMS microbiology ecology, 94(5), fiy042.

Métadonnées additionnelles

Identifiants alternatifs a41ad9bd-cf79-48fa-89a0-102ddf4d4e61
https://ipt.biodiversity.aq/resource?r=lacustrine_cyanobacteria_antarctica