Fig. S1. Three-year time series of CTD-O variables in the study area, Station 18 off Central Chile. The horizontal black line shows the presence of characteristic cold, high-salinity and low-oxygen upwelling-waters (ESSW) during austral spring-summertime. A seasonal oxygen minimum zone (OMZ) related to the ESSW is clearly distinguishable. Arrows indicate the sampling months.


Fig. S2. Three-year time series of nutrients in the study area, Station 18 off Central Chile. The horizontal black line shows the austral spring-summertime (upwelling-favourable seasons). Arrows indicate the sampling months.


Fig. S3. Rarefaction curves for Nitrospina-like 16S rRNA sequence reads obtained in this study by means of pyrosequencing. For comparison purposes, the analyses included both MI_LOCO metagenomic database off Concepción during the wintertime (9/1/2010) and published sequences from Hawaii ocean time series station ALOHA in the North Pacific subtropical gyre (Mincer et al., 2007).


Fig. S4. Alignments of the specific primer region within the Nitrospina-like 16S rRNA gene sequences. The top sequences are the forward primer sequence (A, B) and the reverse complement of the reverse primer sequence (C, D). Nucleotides that match the specific primer region are represented by dots that indicate identical nucleotides. Mismatches are indicated by letters. The alignments were done by using OTUs arisen from this study (in blue) and those available in the databases (members of an OTU share at least 97% sequence identity).


Fig. S5. 16S RT-qPCR and 16S qPCR relationships of Nitrospina-like bacteria. Dashed line show the 16S rRNA : rDNA ratio = 1.


Table S1. Primers used to amplify 16S rRNA and amoA genes coding for ammonia monooxygenase subunit A, specific target groups, amplified product, sequences and annealing temperatures.


Appendix S1. Experimental procedures.

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