Microzooplankton dilution grazing experiments conducted in phytoplankton rich waters, particularly in polar and subpolar seas, often result in calculation of nonsignificant or negative grazing coefficients. We hypothesized that preparation of filtered seawater (FSW) from water containing high biomass of phytoplankton results in release of allelochemicals that inhibit phytoplankton growth, lowering the net growth of phytoplankton in the more diluted treatments. We tested this hypothesis during blooms of Skeletonema marinoi and Phaeocystis pouchetii in a nutrient‐enriched mesocosm in the Raunefjord, Norway. During the S. marinoi bloom, inhibition of phytoplankton growth occurred in the diluted treatments. Simultaneously the concentration of total as well as dissolved polyunsaturated aldehydes (PUAs) was elevated. Passage of the FSW through a carbon cellulose cartridge to remove dissolved organic material reduced, or eliminated, the inhibition. In the early phase of the P. pouchetii bloom that followed the diatom bloom in the mesocosm, PUA concentration was relatively low and the untreated FSW had a less drastic, but often significant, inhibitory effect on phytoplankton growth. Laboratory experiments with cultures of S. marinoi and P. pouchetii confirmed that material present in filtrate prepared from diluted cultures was self‐inhibitory. Many phytoplankters, particularly during late stages of a bloom, produce inhibitory metabolites that may be released during filtration of the relatively large volumes of seawater needed for dilution experiments. Under some conditions, dilution grazing experiments may underestimate phytoplankton growth coefficients and microzooplankton grazing coefficients.

Polyunsaturated aldehydes (PUAs) are released by several diatom species during predation. Besides other attributed activities, these oxylipins can interfere with the reproduction of copepods, important predators of diatoms. While intensive research has been carried out to document the effects of PUAs on copepod reproduction, little is known about the underlying mechanistic aspects of PUA action. Especially PUA uptake and accumulation in copepods has not been addressed to date. To investigate how PUAs are taken up and interfere with the reproduction in copepods we developed a fluorescent probe containing the α,β,γ,δ-unsaturated aldehyde structure element that is essential for the activity of PUAs as well as a set of control probes. We developed incubation and monitoring procedures for adult females of the calanoid copepod Acartia tonsa and show that the PUA derived fluorescent molecular probe selectively accumulates in the gonads of this copepod. In contrast, a saturated aldehyde derived probe of an inactive parent molecule was enriched in the lipid sac. This leads to a model for PUAs' teratogenic mode of action involving accumulation and covalent interaction with nucleophilic moieties in the copepod reproductive tissue. The teratogenic effect of PUAs can therefore be explained by a selective targeting of the molecules into the reproductive tissue of the herbivores, while more lipophilic but otherwise strongly related structures end up in lipid bodies.

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Single-cell investigations of the diatoms Coscinodsicus granii and Thalassiosira pseudonana were performed using laser desorption/ionization (LDI)-MS without the addition of chemical matrices. The unique cell wall architecture of these microalgae, more precisely the biomineralized nanostructured surface, supported the ionization of cellular as well as surface-related metabolites. In model experiments with purified diatom cell walls of eight species C. granii and T. pseudonana proved to promote the ionization of the polymer polyethylene glycol most efficiently. These species were therefore chosen for further experiments. Without any additional workup, living diatom cells can be washed, can be placed on the LDI target and can immediately be profiled using LDI-MS. Characteristic signals arising from the two species were assigned to common metabolites known from diatom metabolism. Among others, chlorophyll, phospholipids and amino acids were detected. Using these fingerprint signals, we were able to perform species-specific MS imaging down to a single-cell resolution of 20 by 20 µm. The larger C. granii cells can be directly visualized, while more than one of the smaller T. pseudonana cells is needed to generate high-quality images. The introduced technique will pave the way toward a chemotyping of phytoplankton that will enable the automated annotation of microalgal species. But also, an assignment of metabolic plasticity on a single-cell level that could answer fundamental questions about plankton diversity is now in reach.

SummaryMolecular probes are widely used tools in chemical biology that allow tracing of bioactive metabolites and selective labeling of proteins and other biomacromolecules. A common structural motif for such probes consists of a reporter that can be attached by copper(I)-catalyzed 1,2,3-triazole formation between terminal alkynes and azides to a reactive headgroup. Here we introduce the synthesis and application of the new thiazole-based, azide-tagged reporter 4-(3-azidopropoxy)-5-(4-bromophenyl)-2-(pyridin-2-yl)thiazole for fluorescence, UV and mass spectrometry (MS) detection. This small fluorescent reporter bears a bromine functionalization facilitating the automated data mining of electrospray ionization MS runs by monitoring for its characteristic isotope signature. We demonstrate the universal utility of the reporter for the detection of an alkyne-modified small molecule by LC–MS and for the visualization of a model protein by in-gel fluorescence. The novel probe advantageously compares with commercially available azide-modified fluorophores and a brominated one. The ease of synthesis, small size, stability, and the universal detection possibilities make it an ideal reporter for activity-based protein profiling and functional metabolic profiling.