1. Wochenbericht AL569

1. Wochenbericht FS Alkor Reise 569, Fahrtabschnitt 02.03.-05.03.202

Assessing marine phytoplankton eco-evolutionary dynamics and physiological responses to environmental change

Anthropogenic emissions of carbon dioxide (CO2) resulting from fossil fuel burning and changes in land use are affecting our marine environment; for example leading to ocean acidification or ocean warming. In order to understand how climate change will affect biological communities we need to understand all levels of biological responses. Community change as a response to environmental drivers are composed of three components: the physiological responses within an organism, described by its phenotypic plasticity or reaction norm and the ecological and evolutionary responses which are associated with changes on the species and genotype level, respectively. Moreover, there may be eco-evolutionary coupling, thus either ecological interactions such as competition that modify evolutionary responses to physico-chemical changes, or evolutionary change that feeds back to change ecological interactions. Here I study for the first time over the long-term (up to 220 generations) how among two competing phytoplankton species the different response types play out, and whether or not coupling of ecological and evolutionary processes can be found. Additionally I investigated the short-term inter- and intraspecific responses of three phytoplankton species to increased CO2 and what role competitive interactions play on the short-term in a two-species ´community´

Selection by higher-order effects of salinity and bacteria on early life-stages of Western Baltic spring-spawning herring

Habitat stratification by abiotic and biotic factors initiates divergence of populations and leads to ecological speciation. In contrast to fully marine waters, the Baltic Sea is stratified by a salinity gradient that strongly affects fish physiology, distribution, diversity and virulence of important marine pathogens. Animals thus face the challenge to simultaneously adapt to the concurrent salinity and cope with the selection imposed by the changing pathogenic virulence. Western Baltic spring-spawning herring (Clupea harengus) migrate to spawning grounds characterized by different salinities to which herring are supposedly adapted. We hypothesized that herring populations do not only have to cope with different salinity levels but that they are simultaneously exposed to higher-order effects that accompany the shifts in salinity, that is induced pathogenicity of Vibrio bacteria in lower saline waters. To experimentally evaluate this, adults of two populations were caught in their spawning grounds and fully reciprocally crossed within and between populations. Larvae were reared at three salinity levels, representing the spawning ground salinity of each of the two populations, or Atlantic salinity conditions resembling the phylogenetic origin of Clupea harengus. In addition, larvae were exposed to a Vibrio spp. infection. Life-history traits and gene expression analysis served as response variables. Herring seem adapted to Baltic Sea conditions and cope better with low saline waters. However, upon a bacterial infection, herring larvae suffer more when kept at lower salinities implying reduced resistance against Vibrio or higher Vibrio virulence. In the context of recent climate change with less saline marine waters in the Baltic Sea, such interactions may constitute key future stressors

Bachelor-MARSYS education cruise in the Baltic Sea Cruise No. AL551

06.03 – 13.03.2021, Kiel (Germany) – Kiel (Germany) BALTEACH -

Eco-Evolutionary Interaction in Competing Phytoplankton: Nutrient Driven Genotype Sorting Likely Explains Dominance Shift and Species Responses to CO2

How ecological and evolutionary processes interact and together determine species and community responses to climate change is poorly understood. We studied long-term dynamics (over approximately 200 asexual generations) in two phytoplankton species, a coccolithophore (Emiliania huxleyi), and a diatom (Chaetoceros affinis), to increased CO2 growing alone, or competing with one another in co-occurrence. To allow for rapid evolutionary responses, the experiment started with a standing genetic variation of nine genotypes in each of the species. Under co-occurrence of both species, we observed a dominance shift from C. affinis to E. huxleyi after about 120 generations in both CO2 treatments, but more pronounced under high CO2. Associated with this shift, we only found weak adaptation to high CO2 in the diatom and none in the coccolithophore in terms of species’ growth rates. In addition, no adaptation to interspecific competition could be observed by comparing the single to the two-species treatments in reciprocal assays, regardless of the CO2 treatment. Nevertheless, highly reproducible genotype sorting left only one genotype remaining for each of the species among all treatments. This strong evolutionary selection coincided with the dominance shift from C. affinis to E. huxleyi. Since all other conditions were kept constant over time, the most parsimonious explanation for the dominance shift is that the strong evolutionary selection was driven by the experimental nutrient conditions, and in turn potentially altered competitive ability of the two species. Thus, observed changes in the simplest possible two-species phytoplankton “community” demonstrated that eco-evolutionary interactions can be critical for predicting community responses to climate change in rapidly dividing organisms such as phytoplankton

Inter- and intraspecific phenotypic plasticity of three phytoplankton species in response to ocean acidification

Phenotypic plasticity describes the phenotypic adjustment of the same genotype to different environmental conditions and is best described by a reaction norm. We focus on the effect of ocean acidification on inter- and intraspecific reaction norms of three globally important phytoplankton species (Emiliania huxleyi, Gephyrocapsa oceanica and Chaetoceros affinis). Despite significant differences in growth rates between the species, they all showed a high potential for phenotypic buffering (similar growth rates between ambient and high CO2 conditions). Only three coccolithophore genotypes showed a reduced growth in high CO2. Diverging responses to high CO2 of single coccolithophore genotypes compared with the respective mean species responses, however, raise the question of whether an extrapolation to the population level is possible from single-genotype experiments. We therefore compared the mean response of all tested genotypes with a total species response comprising the same genotypes, which was not significantly different in the coccolithophores. Assessing species reaction norms to different environmental conditions on short time scale in a genotype-mix could thus reduce sampling effort while increasing predictive power

Temporal variation in ecological and evolutionary contributions to phytoplankton functional shifts

Communities and their functioning are jointly shaped by ecological and evolutionary processes that manifest in diversity shifts of their component species and genotypes. How both processes contribute to community functional change over time is rarely studied. We here repeatedly quantified eco-evolutionary contributions to CO2-driven total abundance and mean cell size changes after short-, mid-, and longer-term (80, 168, and >168d, respectively) in experimental phytoplankton communities. While the CO2-driven changes in total abundance and mean size in the short- and mid-term could be predominantly attributed to ecological shifts, the relative contribution of evolution increased. Over the longer-term, the CO2-effect and underlying eco-evolutionary changes disappeared, while total abundance increased, and mean size decreased significantly independently of CO2. The latter could be presumably attributed to CO2-independent genotype selection which fed back to species composition. In conclusion, ecological changes largely dominated the regulation of environmentally driven phytoplankton functional shifts at first. However, evolutionary changes gained importance with time, and can ultimately feedback on species composition, and thus must be considered when predicting phytoplankton change

Experimentally decomposing phytoplankton community change into ecological and evolutionary contributions

1. Shifts in microbial communities and their functioning in response to environmental change result from contemporary interspecific and intraspecific diversity changes. Interspecific changes are driven by ecological shifts in species composition, while intraspecific changes are here assumed to be dominated by evolutionary shifts in genotype frequency. Quantifying the relative contributions of interspecific and intraspecific diversity shifts to community change thus addresses the essential, yet understudied question as to how important ecological and evolutionary contributions are to total community changes. This debate is to date practically constrained by (a) a lack of studies integrating across organizational levels and (b) a mismatch between data requirements of existing partitioning metrics and the feasibility to collect such data, especially in microscopic organisms like phytoplankton.2. We experimentally assessed the relative ecological and evolutionary contributions to total phytoplankton community changes using a new design and validated its functionality by comparisons to established partitioning metrics. We used a community of coexisting Emiliania huxleyi and Chaetoceros affinis with initially nine genotypes each. First, we exposed the community to elevated CO2 concentration for 80 days (similar to 50 generations) to induce interspecific and intraspecific diversity changes and a total abundance change. Second, we independently manipulated the induced interspecific and intraspecific diversity changes in an assay to quantify the corresponding ecological and evolutionary contributions to the total change. Third, we applied existing partitioning metrics to our experimental data and compared the outcomes.3. Total phytoplankton abundance declined to one-fifth in the high CO2 exposed community compared to ambient conditions. Consistently across all applied partitioning metrics, the abundance decline could predominantly be explained by ecological shifts and to a low extent by evolutionary changes.4. We discuss potential consequences of the observed community changes on ecosystem functioning. Furthermore, we explain that the low evolutionary contributions likely resulted of intraspecific diversity changes that occurred irrespectively of CO2. We discuss how the assay could be upscaled to more realistic settings, including more species and drivers. Overall, the presented calculations of eco-evolutionary contributions to phytoplankton community changes constitute another important step towards understanding future phytoplankton shifts, and eco-evolutionary dynamics in general.</p

Population-specific responses in physiological rates of Emiliania huxleyi to a broad CO2 range

Although coccolithophore physiological responses to CO2-induced changes in seawater carbonate chemistry have been widely studied in the past, there is limited knowledge on the variability of physiological responses between populations from different areas. In the present study, we investigated the specific responses of growth, particulate organic (POC) and inorganic carbon (PIC) production rates of three populations of the coccolithophore Emiliania huxleyi from three regions in the North Atlantic Ocean (Azores: six strains, Canary Islands: five strains, and Norwegian coast near Bergen: six strains) to a CO2 partial pressure (pCO2) range from 120 to 2630 µatm. Physiological rates of each population and individual strain increased with rising pCO2 levels, reached a maximum and declined thereafter. Optimal pCO2 for growth, POC production rates, and tolerance to low pH (i.e., high proton concentration) was significantly higher in an E. huxleyi population isolated from the Norwegian coast than in those isolated near the Azores and Canary Islands. This may be due to the large environmental variability including large pCO2 and pH fluctuations in coastal waters off Bergen compared to the rather stable oceanic conditions at the other two sites. Maximum growth and POC production rates of the Azores and Bergen populations were similar and significantly higher than that of the Canary Islands population. This pattern could be driven by temperature–CO2 interactions where the chosen incubation temperature (16 °C) was slightly below what strains isolated near the Canary Islands normally experience. Our results indicate adaptation of E. huxleyi to their local environmental conditions and the existence of distinct E. huxleyi populations. Within each population, different growth, POC, and PIC production rates at different pCO2 levels indicated strain-specific phenotypic plasticity. Accounting for this variability is important to understand how or whether E. huxleyi might adapt to rising CO2 levels

Gene expression analysis in Atlantic herring (Clupea harengus) in response to temperature and CO2

Major perturbations associated with climate change are decreases in ocean pH levels (ocean acidification, OA) and the increase of mean and variance in temperature. Early life stages such as larval fish may be particularly vulnerable to such stressors either alone or in combination. Earlier studies have established strong phenotypic effects on larval herring such as a decrease in RNA/DNA ratio as a response to hypercapnia or behavioural changes. As an ecologically and economically important fish species studies on the tolerance of Atlantic herring to cope with an altered environment are of prime interest. I quantified gene expression of 32 target genes in larval Atlantic herring and correlated the patterns with identified phenotypic responses to hypercapnia and temperature increase. In a first experiment larvae were kept under three different levels of C02 (380, 1800 and 4200µatm). A second major objective was to address the interaction of OA with temperature which was fully crossed (3°C higher than ambient) with C02 environment (ambient and lOOOppm) in a second experiment. Samples were taken at several time points so as to detect windows of sensitivity to OA and warming stressors. I selected 32 candidate genes putatively involved in acid base regulation, heat stress regulation, immune system response and metabolism. Gene expression levels were quantified using the Dynamic ArrayTM IFC Chip gene expression method (Fluidigm®). When disentangling the different responses and levels of response of Atlantic herring to environmental stress, two different stress responses were detected: hormesis and phenotypic buffering. On the one hand, in response to the three different levels of C02 a dose dependent response - hormesis - could be seen. On the other hand, C02 and temperature together showed a compensatory effect on gene expression level and apparently no effect on the phenotype indicating the larvae's ability to maintain a functioning phenotype in a changed environment which is described by phenotypic buffering. However, the effect of ocean acidification and warming in an environmentally varying coastal habitat in which herring larvae develop with daily fluctuations in temperature and C02 may be underestimated by the levels of C02 of this study