Genome- and transcriptome-assisted development of nuclear insertion/deletion markers for Calanus species (Copepoda: Calanoida) identification

Copepods of the genus Calanus are key zooplankton species in temperate to arctic marine ecosystems. Despite their ecological importance, species identification remains challenging. Furthermore, the recent report of hybrids among Calanus species highlights the need for diagnostic nuclear markers to efficiently identify parental species and hybrids. Using next-generation sequencing analysis of both the genome and transcriptome from two sibling species, Calanus finmarchicus and Calanus glacialis, we developed a panel of 12 nuclear insertion/deletion markers. All the markers showed species-specific amplicon length. Furthermore, most of the markers were successfully amplified in other Calanus species, allowing the molecular identification of Calanus helgolandicus, Calanus hyperboreus and Calanus marshallae.

PhytoREF: a reference database of the plastidial 16S rRNA gene of photosynthetic eukaryotes with curated taxonomy

Photosynthetic eukaryotes have a critical role as the main producers in most ecosystems of the biosphere. The ongoing environmental metabarcoding revolution opens the perspective for holistic ecosystems biological studies of these organisms, in particular the unicellular microalgae that often lack distinctive morphological characters and have complex life cycles. To interpret environmental sequences, metabarcoding necessarily relies on taxonomically curated databases containing reference sequences of the targeted gene (or barcode) from identified organisms. To date, no such reference framework exists for photosynthetic eukaryotes. In this study, we built the PhytoREF database that contains 6490 plastidial 16S rDNA reference sequences that originate from a large diversity of eukaryotes representing all known major photosynthetic lineages. We compiled 3333 amplicon sequences available from public databases and 879 sequences extracted from plastidial genomes, and generated 411 novel sequences from cultured marine microalgal strains belonging to different eukaryotic lineages. A total of 1867 environmental Sanger 16S rDNA sequences were also included in the database. Stringent quality filtering and a phylogeny-based taxonomic classification were applied for each 16S rDNA sequence. The database mainly focuses on marine microalgae, but sequences from land plants (representing half of the PhytoREF sequences) and freshwater taxa were also included to broaden the applicability of PhytoREF to different aquatic and terrestrial habitats. PhytoREF, accessible via a web interface (http://phytoref.fr), is a new resource in molecular ecology to foster the discovery, assessment and monitoring of the diversity of photosynthetic eukaryotes using high-throughput sequencing.

PhytoREF: a reference database of the plastidial 16S rRNA gene of photosynthetic eukaryotes with curated taxonomy

Photosynthetic eukaryotes have a critical role as the main producers in most ecosystems of the biosphere. The ongoing environmental metabarcoding revolution opens the perspective for holistic ecosystems biological studies of these organisms, in particular the unicellular microalgae that often lack distinctive morphological characters and have complex life cycles. To interpret environmental sequences, metabarcoding necessarily relies on taxonomically curated databases containing reference sequences of the targeted gene (or barcode) from identified organisms. To date, no such reference framework exists for photosynthetic eukaryotes. In this study, we built the PhytoREF database that contains 6490 plastidial 16S rDNA reference sequences that originate from a large diversity of eukaryotes representing all known major photosynthetic lineages. We compiled 3333 amplicon sequences available from public databases and 879 sequences extracted from plastidial genomes, and generated 411 novel sequences from cultured marine microalgal strains belonging to different eukaryotic lineages. A total of 1867 environmental Sanger 16S rDNA sequences were also included in the database. Stringent quality filtering and a phylogeny-based taxonomic classification were applied for each 16S rDNA sequence. The database mainly focuses on marine microalgae, but sequences from land plants (representing half of the PhytoREF sequences) and freshwater taxa were also included to broaden the applicability of PhytoREF to different aquatic and terrestrial habitats. PhytoREF, accessible via a web interface (http://phytoref.fr), is a new resource in molecular ecology to foster the discovery, assessment and monitoring of the diversity of photosynthetic eukaryotes using high-throughput sequencing.

Molecular analysis of Continuous Plankton Recorder samples, an examination of echinoderm larvae in the North Sea

Analysis of the biological time series of plankton samples collected by the Continuous Plankton Recorder (CPR) in the North Atlantic and North Sea has shown a regime shift in the plankton in this region. Both the distributions of planktonic organisms and their timing of occurrence in the seasonal cycle have changed and these changes appear to ref lect global warming. In the North Sea the planktonic larvae of echinoderms have shown a recent dramatic increase in both relative and absolute abundance and their seasonal peak of occurrence has advanced by 47 days. The identity of the echinoderm larvae involved in this change has, however, remained equivocal. The small size of many organisms like echinoderm larvae combined with incomplete taxonomic keys hinders their visual identification and their fragility often means that useful morphological features are damaged during sampling by the CPR. Here, using new molecular methods applied to CPR samples, we show that planktonic larvae of the benthic Echinocardium cordatum dominate the North Sea plankton. We argue that since this species benefits from mild winters and warmer waters their numerical increase in the plankton is consistent with recent climatic changes that appear to be affecting the wider ecology of this region.

PCR from the CPR offers a historical perspective on marine population ecology.

The Continuous Plankton Recorder (CPR) survey has collected plankton samples from regular tracks across the world's oceans for almost 70 y. Over 299,000 spatially extensive CPR samples are archived and stored in buffered formalin. This CPR archive offers huge potential to study changes in marine communities using molecular data from a period when marine pollution, exploitation and global anthropogenic impact were much less pronounced. However, to harness the amount of data available within the CPR archive fully, it is necessary to improve techniques of larval identification, to genus and species preferably, and to obtain genetic information for historical studies of population ecology. To increase the potential of the CPR database this paper describes the first extraction, amplification by the polymerase chain reaction and utilization of a DNA sequence (mitochondrial 16S rDNA) from a CPR sample, a formalin fixed larval sandeel.