The cosmopolitan distribution of the marine planktonic diatom Chaetoceros socialis Lauder is generally interpreted as a case of wide ecological plasticity. However, cryptic diversity has recently been observed within this taxon and this genetic diversity could extend to phenotypic differences between allopatric populations. Monoclonal strains of C. socialis isolated from NE Atlantic/Arctic and Tyrrhenian Sea waters were cultivated at 2.5, 8 and 13°C, and analysed for morphological, phylogenetic (LSU rRNA gene) and physiological (growth rate, photosynthetic yield) traits. The molecular analysis indicated an unequivocal divergence between the southern and the northern strains. Clear differences in spore morphology between the two groups were also observed, despite their morphological similarity in vegetative form. The physiological data demonstrated a functional partition between the northern and southern strains. The northern strains showed significantly higher growth rates than the southern ones at the lower temperatures, and vice versa at 13°C. Differences between the groups were also observed in the photosynthetic yields. These results reveal the expression of allopatric speciation in several phenotypic characters, providing a functional dimension to the cryptic, or rather pseudo-cryptic, diversity observed within C. socialis. We believe that such an integrated approach is useful for robust taxonomic and functional delimitations of presumed cosmopolitan microorganisms.

In this study, we investigate how metabolic fingerprints are related to temperature. Six common northern temperate diatoms (Attheya longicornis, Chaetoceros socialis, Chaetoceros furcellatus, Porosira glacialis, Skeletonema marinoi, and Thalassiosira gravida) were cultivated at two different temperatures, 0.5 and 8.5 °C. To exclude metabolic variations due to differences in growth rates, the growth rates were kept similar by performing the experiments under light limited conditions but in exponential growth phase. Growth rates and maximum quantum yield of photosynthesis were measured and interpreted as physiological variables, and metabolic fingerprints were acquired by high-resolution mass spectrometry. The chemical diversity varied substantially between the two temperatures for the tested species, ranging from 31% similarity for C. furcellatus and P. glacialis to 81% similarity for A. longicornis. The chemical diversity was generally highest at the lowest temperature.

During normal sample preparation, storage in freezers and subsequent freeze/thaw cycles are commonly introduced. The effect of freeze/thaw cycles on the metabolic profiling of microalgal extracts using HR-MS was investigated. Methanolic extracts of monocultures of Arctic marine diatoms were analyzed immediately after extraction, after seven days of storage at −78 °C (one freeze/thaw cycle), and after additional seven days at −20 °C (two freeze/thaw cycles). Repeated direct infusion high-resolution mass spectrometry analysis of microalgae extracts of the same sample showed that reproducibility was ca. 90% when a fresh (unfrozen) sample was analyzed. The overall reproducibility decreased further by ca. 10% after the first freeze/thaw-cycle, and after one more freeze/thaw cycle the reproducibility decreased further by ca. 7%. The decrease in reproducibility after freeze-thaw cycles could be attributed to sample degradation and not to instrument variability.