Distinct relationships between fluorescence in situ hybridization and 16S rRNA gene- and amplicon-based sequencing data of bacterioplankton lineages.

Catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) and amplicon sequencing of the total (16S rRNA gene) and potentially active (16S rRNA transcripts), community are the major state of the art approaches for assessing the composition of bacterial communities in marine pelagic and other ecosystems. However, CARD-FISH and amplicon sequencing methods have not yet been directly compared to assess the composition of bacterioplankton communities. Therefore, these approaches were used to study the composition of bacterial communities in two North Sea seawater mesocosm experiments supplemented with diatom-derived organic matter (OM). All approaches revealed Proteobacteria and Bacteroidetes as major components of the bacterioplankton communities. The Roseobacter group and its RCA cluster, as well as Bacteroidetes and Gammaproteobacteria, responded most strongly to OM addition, whereas the SAR11 clade responded in only one of the two mesocosms. A correlation analysis showed that CARD-FISH and amplicon sequencing data of the SAR11 clade and the Roseobacter group, together with its RCA cluster, were highly significantly correlated, whereas Bacteroidetes did not yield any significant correlation and Gammaproteobacteria was only correlated with the potentially active fraction. However, subgroups of these phylogenetic groups, the SAR92 clade, the genera Pseudoalteromonas and Polaribacter, exhibited significant correlations in one of the two mesocosms. Correlations of CARD-FISH with amplicon sequencing data from the total and potentially active fractions of these lineages exhibited distinct differences. The study showed that CARD-FISH and amplicon sequencing data of distinct bacterioplankton groups and especially the phylogenetic lineages at a higher taxonomic level were correlated but reflected different aspects of their growth dynamics.