ALBERT

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 understand the thermal plasticity of a coastal foundation species across its latitudinal distribution, we assess physiological responses to high temperature stress in the kelp Laminaria digitata in combination with population genetic characteristics and relate heat resilience to genetic features and phylogeography. We hypothesize that populations from Arctic and cold-temperate locations are less heat resilient than populations from warm distributional edges. Using meristems of natural L. digitata populations from six locations ranging between Kongsfjorden, Spitsbergen (79°N), and Quiberon, France (47°N), we performed a common-garden heat stress experi- ment applying 15°C to 23°C over eight days. We assessed growth, photosynthetic quantum yield, carbon and nitrogen storage, and xanthophyll pigment contents as response traits. Population connectivity and genetic diversity were analyzed with microsatellite markers. Results from the heat stress experiment suggest that the upper temperature limit of L. digitata is nearly identical across its distribution range, but subtle differences in growth and stress responses were revealed for three popu- lations from the species’ ecological range margins. Two populations at the species’ warm distribution limit showed higher temperature tolerance compared to other populations in growth at 19°C and recovery from 21°C (Quiberon, France), and pho- tosynthetic quantum yield and xanthophyll pigment responses at 23°C (Helgoland, Germany). In L. digitata from the northernmost population (Spitsbergen, Norway), quantum yield indicated the highest heat sensitivity. Microsatellite genotyping revealed all sampled populations to be genetically distinct, with a strong hierarchical structure between southern and northern clades. Genetic diversity was lowest in the isolated population of the North Sea island of Helgoland and highest in Roscoff in the English Channel. All together, these results support the hypothesis of moderate local differentiation across L. digitata's European distribution, whereas effects are likely too weak to ameliorate the species’ capacity to withstand ocean warming and marine heatwaves at the southern range edge.

Description: The plasticity of different kelp populations to heat stress has seldom been investigated excluding environmental effects due to thermal histories, by raising a generation under common garden conditions. Comparisons of populations in the absence of environmental effects allow unbiased quantification of the meta-population adaptive potential and resolution of population-specific differentiation. Following this approach, we tested the hypothesis that genetically distinct arctic and temperate kelp exhibit different thermal phenotypes, by comparing the capacity of their microscopic life stages to recover from elevated temperatures. Gametophytes of Laminaria digitata (Arctic and North Sea) grown at 15˚C for 3 years were subjected to common garden conditions with static or dynamic (i.e., gradual) thermal treatments ranging between 15 and 25˚C and also to darkness. Gametophyte growth and survival during thermal stress conditions, and subsequent sporophyte recruitment at two recovery temperatures (5 and 15˚C), were investigated. Population-specific responses were apparent; North Sea gametophytes exhibited higher growth rates and greater sporophyte recruitment than those from the Arctic when recovering from high temperatures, revealing differential thermal adaptation. All gametophytes performed poorly after recovery from a static 8-day exposure at 22.5˚C compared to the response under a dynamic thermal treatment with a peak temperature of 25˚C, demonstrating the importance of gradual warming and/or acclimation time in modifying thermal limits. Recovery temperature markedly affected the capacity of gametophytes to reproduce following high temperatures, regardless of the population. Recovery at 5˚C resulted in higher sporophyte production following a 15˚C and 20˚C static exposure, whereas recovery at 15˚C was better for gametophyte exposures to static 22.5˚C or dynamic heat stress to 25˚C. The subtle performance differences between populations originating from sites with contrasting local in situ temperatures support our hypothesis that their thermal plasticity has diverged over evolutionary time scales.

Description: Kelp beds worldwide are under pressure from ongoing climate and environmental change. Along European coastlines increases in seawater temperature and changes in nutrient conditions occur where upwelling events are disrupted and also along eutrophicated coasts. In addition, seaweed responses to change may interact with seasonal daylength cycles. We performed a factorial experiment to examine the combined effects of seawater temperatures, nutrient regimes, and photoperiod (long and short days) in order to better understand how latitudinal or seasonal differences in daylengths affects the sensitivity of transient microscopic kelp stages of Laminaria digitata from the North Sea, to warming and eutrophication. While the optimal temperature range for vegetative gametophyte growth was 10-18ºC under long summer photoperiod conditions, gametogenesis was induced at lower temperatures between 5 and 15ºC, with maximum sporophyte development under long photoperiods and enriched nutrient regimes, which represents local late spring conditions. Although gametogenesis was fastest at 10-15ºC, sporophyte recruitment was highest at 5ºC. As these particular early life cycle processes in L. digitata have different temperature optima, this may drive the seasonal cycle of recruitment in the field. Increasing summer temperatures due to global warming will increase gametophyte size due to enhanced vegetative growth and inhibition of gametogenesis. This will probably lead to delayed but enhanced recruitment of new sporophytes under cooler autumn to spring conditions over a wide geographical scale, preventing the formation of juvenile sporophytes under stressful summer conditions and possibly changing annual recruitment patterns.

Description: Kelp forests are important habitats in the strongly environmentally and seasonally variable Arctic. There is a critical lack of knowledge about how seasonal conditions and climate change scenarios influence survival and reproduction of kelp early life stages. To better understand the regulation of kelp life cycle processes in this harsh environment we focused on the physiological performance and reproductive success of early life stages in Alaria esculenta and Laminaria digitata from Kongsfjorden, Spitsbergen. Gametophyte growth and survival during Arctic winter and subsequent sporophyte recruitment under spring conditions were investigated. Winter conditions (2°C, complete darkness) halted gametophyte growth and prevented the onset of gametogenesis in both species. The gametophytes of L. digitata but not A. esculenta became fertile after returning to spring conditions, suggesting that sporogenesis, sexual reproduction and recruitment in A. esculenta must occur successively during summer/autumn while in L. digitata a new generation of sporophytes could develop from overwintering gametophytes. The effects of simulated canopy shading (offering protection against extreme irradiance stress, particularly as sea ice retreats), present-day and projected Arctic summer seawater temperatures, and nutrient levels on gametophyte survival, fertility and sporophyte recruitment success were also investigated in both species. A. esculenta gametophytes had greater survival and reproductive success than L. digitata, except under very low light (simulating dense canopy). In contrast, shading was required for reproductive success in L. digitata gametophytes. Predicted summer temperatures of 9°C reduced sexual reproduction in both species. Interactions observed between these environmental drivers probably reflect species-specific seasonal patterns of survival and reproduction. These differences between kelp species in response to abiotic factors and light levels (simulated canopy shading) suggest that climate change could alter community structure in the Arctic through effects on sexual reproduction and sporophyte recruitment success.

Description: Marine forests and kelps as their foundation species are threatened by ocean warming especially at the warm distributional edges. Previously identified genetic divergence and ecotypic differentiation within kelp species may allow to produce more resilient lineages by intraspecific outbreeding among populations. 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. The upper survival temperature of microscopic sporophytes was lower for the inbred Arctic selfing (21°C) than for the temperate selfing and the reciprocal crosses (22°C). Only in the temperate selfing, 4.5% of sporophytes survived 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 gene expression in addition to physiological parameters. Growth and optimum quantum yield decreased similarly in the reciprocal crosses and the temperate selfing at 19 and 20.5°C, while inbred Arctic sporophytes died within seven days at both 19 and 20.5°C. In response to 20.5°C, 252 genes were constitutively regulated across all surviving lineages, which we use to describe metabolic regulation patterns in response to heat stress in kelp. At sublethal 20.5°C, ca. 150 genes were differentially expressed by either crossed lineage in comparison to the temperate selfing, indicating that they maintained a growth response similar to the temperate selfing with differential metabolic regulation during sublethal heat stress. Subtle differences in physiology and the differential expression of nine genes between the reciprocal crosses at 20.5°C indicate that female and male gametophytes may contribute differently to offspring traits. We consider potential inbreeding depression in the Spitsbergen selfing and quantify the better performance of both crosses using heterosis-related parameters. We discuss the potential and risks of outbreeding to produce more resilient crops for mariculture and marine forest restoration.

Description: In recent years, kelp populations worldwide have faced decline and extirpation at their equatorward limits, while models predict a poleward shift of kelp ecosystems during climate change. To gain an understanding of local thermal adaptation and response plasticity in a forest-forming kelp species, we assessed populations of Laminaria digitata along its entire European distribution range for their capacity to withstand high temperature stress, and analysed population structure and diversity with microsatellite markers (n=12). We sampled wild meristematic L. digitata material (n=30) at six locations ranging from Kongsfjorden, Spitsbergen, to the southernmost distribution limit in Quiberon, France. In a heatwave experiment, we subjected samples from all locations to the same, sublethal temperature treatments (15–23°C for eight days including acclimation) and assessed growth, storage compounds, photosynthetic efficiency and pigment contents as response traits. Recovery was assessed following seven days at 15°C. Microsatellite genotyping revealed all sampled populations to be genetically distinct entities, underlying strong regional structuring between southern and northern clades. Genetic diversity was highest at the southern distribution limit in Quiberon and lowest in the geographically isolated population on the island of Helgoland in the North Sea. The physiological response of L. digitata to temperature was similar over the entire distribution range and did not reflect the mean temperature gradient along the latitudinal gradient. However, material from Spitsbergen and Helgoland presented subtle differentiations in their temperature responses, which reflect long-term local temperature histories at these sites. Finally, a heatwave reaching 23°C for five days led to a cessation of growth, from which none of the sampled populations recovered. Our results suggest that the heat stress response of L. digitata is generally stable across its distribution range, despite strong genetic structuring of the populations. Slight local differentiation occurred in populations from the most distinct thermal environments, but 23°C posed a growth limit for all populations. This implies that local adaptation in trailing edge populations of L. digitata might not alleviate detrimental effects of global warming.

Description: In haplodiplontic lineages, sexual reproduction occurs in haploid parents without meiosis. Although widespread in multicellular lineages such as brown algae (Phaeophyceae), haplodiplontic gametogenesis has been little studied at the molecular level. We addressed this by generating an annotated reference transcriptome for the gametophytic phase of the sugar kelp, Saccharina latissima. Transcriptional profiles of microscopic male and female gametophytes were analysed at four time points during the transition from vegetative growth to gametogenesis. Gametogenic signals resulting from a switch in culture irradiance from red to white light activated a core set of genes in a sex-independent manner, involving rapid activation of ribosome biogenesis, transcription and translation related pathways, with several acting at the post-transcriptional or post-translational level. Additional genes regulating nutrient acquisition and key carbohydrate-energy pathways were also identified. Candidate sex-biased genes under gametogenic conditions had potentially key roles in controlling female- and male-specific gametogenesis. Among these were several sex-biased or -specific E3 ubiquitin-protein ligases that may have important regulatory roles. Females specifically expressed several genes that coordinate gene expression and/or protein degradation, and the synthesis of inositol-containing compounds. Other female-biased genes supported parallels with oogenesis in divergent multicellular lineages, in particular reactive oxygen signalling via an NADPH-oxidase. Males specifically expressed the hypothesised brown algal sex-determining factor. Male-biased expression mainly involved upregulation of genes that control mitotic cell proliferation and spermatogenesis in other systems, as well as multiple flagella-related genes. Our data and results enhance genome-level understanding of gametogenesis in this ecologically and economically important multicellular lineage.

Description: Many organisms have endogenous clocks that synchronize biological processes with environmental changes, leading to optimized development and reproduction. However, certain environments, like the Arctic, pose a special challenge to circadian clocks, particularly due to extreme seasonal changes in daylength, ranging from permanent sunlight to complete darkness. Kelps seem to be well adapted to the variable environmental conditions characteristic of this region. However, daylength might affect kelp species that use circadian rhythms to control the timing of daily egg release from female gametophytes. We aimed to investigate how daylength and light intensity affect gametogenesis and reproductive success of summer-reproducing kelp species (using Alaria esculenta as a model). As daylength and temperature co-vary most of the year, we also investigated the thermal resilience of the sporophytes developed under different daylengths to understand if there is a cross-tolerance between light doses and temperature tolerance. Although continuous daylight, characteristic of Arctic summers, enhanced gametogenesis and increased gametophyte vegetative growth, and thereby the number of potential reproductive gametophyte cells, sporophyte production was higher under long (16 h light:8 h dark) and intermediate (12:12 h) days. Sporophyte growth was triggered by changing daylength from short to long days, suggesting a synchronization with annual daylength variation. High daily light doses during reproduction and early development improved subsequent sporophyte survival at high (sub)lethal temperatures, indicating cross-tolerance between light and temperature. Reproductive success in Arctic A. esculenta was hampered under continuous light, and we hypothesize that this might result from disturbance of synchronized egg release and subsequent fertilization.