QICS Data: Effect of a controlled sub-seabed release of CO2 in Ardmucknish Bay on microbial activity (2012 - 2014)

Sediment samples were collected 7 days before the CO2 supply was turned on (D-7/8), D13/14 and D35/36 during CO2 release, and after the CO2 release ended on D41/42, D55/56 and D127/128 at each of four zones (zone 1 = at epicentre; zone 2 (25m distant), zone 3 (75m distant), and zone 4 (control, 450m distant). At each zone, five replicate sediment samples were manually collected by divers using five 50 ml syringes with the end cut off. Each syringe was sealed with a rubber bung and transported back to the lab were the top 1 cm was sectioned and frozen at -80 °C. RNA and DNA were extracted from 2 g sediment using the MoBio RNA PowerSoil® Total RNA Isolation Kit with the DNA elution accessory kit according to the manufacturer’s instructions. RNA was reverse transcribed using the QuantiTect Reverse Transcription Kit (Qiagen) with 0.2 μg of RNA and the supplied random primers. An ABI 7000 sequence detection system (Applied Biosystems, Foster City, USA) and QuantiFast SYBR Green PCR Kit (Qiagen) was used for all qPCR measurements. The abundance of bacterial, archaeal and cyanobacterial/chloroplast 16S rRNA (from cDNA) and genes (from DNA) in each sediment sample was determined using quantitative PCR. The method is described in detail in Tait et al. 2015. In addition, for each sediment sample, the abundance of Thaumarchaeal and bacterial amoA transcripts and genes, nitrite reducer (nirS) and hydrazine oxidoreductase (hzo) genes and transcripts was determined. The method is described in detail in Watanabe et al. 2015. The data in the table QICS microbial qPCR data.txt is in the format of Dx (relative to day of injection). D-7/8: 10/11 May 2012 D13/14: 30/31 May 2012 D35/D36: 20/21 June 2012 D41/D42: 27/28 June 2012 D54/D55: 9 July/10 July 2012 D127/D128: 19/20 September 2012 Zones are as described above and data is shown for the 5 replicate cores. Numbers are 16S rRNA/transcript/gene copies per gram sediment. DGGE analysis of cyanobacteria/chloroplast 16S rRNA (cDNA) sequences was performed with the INGENYphorU DGGE system (Ingeny, The Netherlands), as described in detail in Tait et al. 2015 . Replicate cores were combined to give a total of 24 PCR products. The original gel image is QICS cyanobacteria 16S rRNA DGGE image.tif. Gel Compar II software was used to analyse DGGE fingerprinting patterns (Applied Maths, Netherlands) and the data imported into PRIMER vs. 6.1 multivariate analysis software (Clarke and Gorley 2006) for statistical analysis. The data in the table QICS cyanobacteria 16S rRNA DGGE data.txt contains relative abundance of band density for each sample. Data is in the format of Dx as above. Bands are numbered according to their distance migrated down the gel. To compare bacterial communities at each zone and over time, 16S rRNA fragments were amplified and analysed using terminal restriction length polymorphism (T-RFLP) using the protocol of Laverock et al. (2010). 6-FAM labelled PCR products were digested with the restriction enzyme AluI, and the fragment profile for each sample determined by DNA Sequencing Services (Dundee University). Resulting electropherograms were initially analysed using GeneMapper 3.7 software (Applied Biosystems) and subsequently using T-Align (Smith et al., 2005). This data can be found in Blackford et al. (2014) Nature Climate Change DOI: 10.1038/NCLIMATE2381. Relative abundance of TRFs for each sample can be found in QICS bacterial 16S rRNA T-RFLP data.txt. Again, the data is in the format of Dx as above. TRFs are numbered according to their nucleotide length. The relative abundance and composition of 16S rRNA (cDNA) in the five replicate samples collected from zones 1 and zones 4 on D-7/8, D13/14, D35/36 and D41/42 were determined using 16S rRNA tagged pyrosequencing, as described in Tait et al. 2015 . Sequence data has been archived at the European Nucleotide Archive, accession number PRJEB7642.

INSPIRE Implementing rules laying down technical arrangements for the interoperability and harmonisation of Geology

Commission Regulation (EU) No 1089/2010 of 23 November 2010 implementing Directive 2007/2/EC of the European Parliament and of the Council as regards interoperability of spatial data sets and services