ALBERT

Description: Highlights: • Acclimation of Fucus vesiculosus under a highly resolved temperature gradient. • Fucus vesiculosus exhibits a broad temperature optimum for growth from 10 to 24 °C. • Upper survival temperature of Fucus vesiculosus is between 26 and 27 °C. • Optimal temperature for photosynthesis is higher compared to that for growth. • Fucus in the Baltic Sea may become a “loser” under increasing temperatures. Abstract: Seaweeds provide important ecosystem services in coastal areas, and loss of these macrophytes due to anthropogenic global change and warming is a worldwide concern. Fucus vesiculosus L. (Phaeophyceae) is the most abundant and hence ecologically most important primary producer, carbon sink and habitat provider in the western Baltic Sea. Therefore, we used this keystone species to test phenotypic acclimation of physiological performance traits (growth, photosynthesis and metabolites) of F. vesiculosus apices in a well-defined and highly resolved temperature gradient (5–29 °C), supported by highly temporally resolved measurements. Temperature requirements of growth and photosynthesis were evaluated in three weeks exposure experiments, and changing tolerance ranges for survival over time were determined. Fucus vesiculosus was able to grow and survive over a temperature range from 5 to 26 °C without any injury or visible damage of the apical growing meristem over all three weeks. However, at higher water temperatures (≥ 27 °C) growth rapidly decreased from day three onwards and progressive necrosis was observed at 28 and 29 °C. Stress-induced decrease in growth rate was already indicated by the effective quantum yield of chlorophyll fluorescence of photosystem II (PSII) several days in advance. Optimal temperature for photosynthesis (24 °C), measured as electron transport rate, was higher compared to that for growth (15–20 °C). Accordingly, the concentration of mannitol, the main product of photosynthesis, increased with higher temperatures. Understanding physiological responses of keystone macroalgae with respect to temperature and time is important, because rising global temperatures and summer heat wave frequencies and duration may affect the ecological functions of F. vesiculosus in the western Baltic Sea.

Description: Fucus vesiculosus L. (Phaeophyceae) is the most abundant and hence ecologically most important primary producer, carbon sink and habitat provider in the western Baltic Sea. All F. vesiculosus L. specimens were collected on 23 April 2014 from a depth of 0.2-1 m in the non-tidal Kiel Fjord, western Baltic Sea (54°27'N; 10°12'E), where this species forms dense and almost monospecific stands on stones. After sampling the algal thalli were stored in a refrigerator box with water from the sampling site, transported to Bremerhaven and stored at 10 °C for one day in filtered seawater. Experiments were conducted with vegetative apical tips (6.7±0.5 cm length), the actively growing region of F. vesiculosus, which were randomly selected and cut from 144 different individuals prior to the experiments. These tips were acclimated to laboratory conditions for three days in filtered seawater at 10 °C before the start of the experiment. Furthermore, 30 additional vegetative apices were freeze-dried to document the initial biochemical status of F. vesiculosus in its native habitat. A temperature gradient was installed in a walk-in constant cooling chamber (15 °C) in nine water baths (5, 10, 15, 20, 24, 26, 27, 28 and 29 °C ± 0.1 °C) which were tempered by thermostats (5, 10 and 15 °C: Huber Variostat CC + Pilot ONE, Peter Huber Kältemaschinen GmbH, Offenburg, Germany; 20 and 28 °C: Haake DC3, Thermo Fisher Scientific Inc., Waltham, USA; 24, 26, 27 and 29 °C: Haake DC10). Every temperature treatment consisted of four 2 L glass beakers (n = 4). In each beaker four F. vesiculosus apices were grown in 2 µm-filtered North Sea water diluted with demineralized water in a ratio of 1:1 and enriched with nutrients after Provasoli (1968; 1/10 enrichment), leading to a salinity of about 15.6 which equaled habitat conditions. The algae were exposed to an irradiance of 130 µmol photons m-2 s-1 ±10 % (Powerstar HGI-TS 150 W, OSRAM GmbH, Bad Homburg, Germany) measured at the top of the beaker under a 16:8 h L:D cycle. The media in the beakers was changed every third or fourth day and aerated with artificial air containing 380 ppm CO2 (gas mixing device; HTK Hamburg GmbH, Hamburg, Germany). Before the experiment, the algae were acclimated to the final temperatures in steps of 5 °C for 2 days each, beginning at 10 °C. After 21 days exposure time, three out of four samples per replicate were freeze-dried for further biochemical analyses, and afterwards the thermostats were turned off to reduce the temperature to 16±0.4 °C for another 10 days permitting growth under post-culture conditions.

Description: To assess the thermal adaptation of microscopic stages of the kelp Laminaria digitata along latitudes, we conducted laboratory experiments on samples from six locations in the NE Atlantic (Spitsbergen (SPT), Tromsø (TRM), Bodø (BOD; all Norway), Helgoland (HLG; Germany), Roscoff (ROS) and Quiberon (QUI; both France)), spanning the species' entire distribution range. In experiment 1, we exposed gametophytes to (sub-) lethal high priming temperatures (20-25°C) for two weeks, followed by two weeks of recovery at 15°C, to observe gametophyte survival and sporophyte formation. In experiment 2, samples were subjected to (sub-) optimal low temperatures (0-15°C) for 21 days, to assess gametophyte survival, sporophyte formation and growth. During the experiments, samples were kept in 15 µmol photons/m²/s white light under a 16:8h light:dark cycle. Prior to the experiments, cultures were stored at 15°C in iron-free ½ Provasoli enriched seawater in 3-4 µmol photons/m²/s red light.

Description: In a mechanistic investigation of heat stress, heterosis (hybrid vigour), and underlying gene expression patterns, we assessed the thermal performance of inbred (selfings) and outbred (reciprocal crosses) sporophytes of the N-Atlantic kelp Laminaria digitata among clonal isolates from two divergent populations; one from the temperate North Sea (Helgoland) and one from the Arctic (Spitsbergen). First, we investigated the upper thermal tolerance of microscopic sporophytes in a 14-day experiment applying sublethal to lethal 20–23°C. We then subjected 4–7 cm long sporophytes to a control temperature (10°C), moderate (19°C) and sublethal to lethal heat stress (20.5°C) for 18 days to assess the physiological parameters growth and optimum quantum yield.

Description: To assess the thermal adaptation of microscopic stages of the kelp Laminaria digitata along latitudes, we conducted laboratory experiments on samples from six locations in the NE Atlantic (Spitsbergen (SPT), Tromsø (TRM), Bodø (BOD; all Norway), Helgoland (HLG; Germany), Roscoff (ROS) and Quiberon (QUI; both France)), spanning the species' entire distribution range. Gametophyte stock cultures from the Alfred Wegener Institute, Helmholtz Centre for Polar and Marine Research were used. Prior to the experiments, cultures were stored at 15°C in iron-free ½ Provasoli enriched seawater in 3-4 µmol photons/m²/s red light. In experiment 1, we exposed gametophytes to (sub-) lethal high priming temperatures (20-25°C) for two weeks, followed by two weeks of recovery at 15°C, to observe gametophyte survival and sporophyte formation. During the experiments, samples were kept in 15 µmol photons/m²/s white light under a 16:8h light:dark cycle.

Description: To assess the thermal adaptation of microscopic stages of the kelp Laminaria digitata along latitudes, we conducted laboratory experiments on samples from six locations in the NE Atlantic (Spitsbergen (SPT), Tromsø (TRM), Bodø (BOD; all Norway), Helgoland (HLG; Germany), Roscoff (ROS) and Quiberon (QUI; both France)), spanning the species' entire distribution range. Gametophyte stock cultures from the Alfred Wegener Institute, Helmholtz Centre for Polar and Marine Research were used. Prior to the experiments, cultures were stored at 15°C in iron-free ½ Provasoli enriched seawater in 3-4 µmol photons/m²/s red light. In experiment 2, samples were subjected to (sub-) optimal low temperatures (0-15°C) for 21 days, to assess gametophyte survival, sporophyte formation and growth. During the experiments, samples were kept in 15 µmol photons/m²/s white light under a 16:8h light:dark cycle. Sporophyte growth rates both in length and in width were determined as follows: GR = (x2-x1)/(t2-t1), where x is the length or width (μm) and t is the time in weeks at time point 1 and 2.

Description: To assess the thermal adaptation of microscopic stages of the kelp Laminaria digitata along latitudes, we conducted laboratory experiments on samples from six locations in the NE Atlantic (Spitsbergen (SPT), Tromsø (TRM), Bodø (BOD; all Norway), Helgoland (HLG; Germany), Roscoff (ROS) and Quiberon (QUI; both France)), spanning the species' entire distribution range. Gametophyte stock cultures from the Alfred Wegener Institute, Helmholtz Centre for Polar and Marine Research were used. Prior to the experiments, cultures were stored at 15°C in iron-free ½ Provasoli enriched seawater in 3-4 µmol photons/m²/s red light. In experiment 1, we exposed gametophytes to (sub-) lethal high priming temperatures (20-25°C) for two weeks, followed by two weeks of recovery at 15°C, to observe gametophyte survival and sporophyte formation. During the experiments, samples were kept in 15 µmol photons/m²/s white light under a 16:8h light:dark cycle.

Description: We investigated effects of temperature during ontogeny of the kelp Laminaria digitata across haploid gametophyte and diploid sporophyte life cycle stages in 5 distinct genetic lines. We sampled meristematic discs from wild sporophytes on the island of Helgoland (North Sea; 54.1779 N, 7.8926 E) in May 2017, July 2017 and February 2018 and conducted a seasonal growth experiment at 5°C and 15°C over 14 days (experiment 1). Based on meiospores from five individuals (genetic lines) sampled in July 2017, we applied a full-factorial experimental design to generate different temperature histories by applying 5°C and 15°C during meiospore germination, gametogenesis of parental gametophytes and recruitment of offspring sporophytes (19-26 days; recruitment), and juvenile sporophyte rearing (91-122 days). We then tested for thermal plasticity among temperature history treatments at 5°C and 15°C in a final 12-day experiment assessing growth, the storage compound mannitol, carbon and nitrogen contents, and fluorometric responses in 3-4 month old sporophytes (experiment 2).