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De Clerck, O.

Publications and source records attributed to De Clerck, O..

15 recordsLinked to original sources

Beyond establishment: incorporating physiological performance into predictions of invasion risk

Biological invasions are a major driver of global change, reshaping ecosystems and threatening biodiversity worldwide. Anticipating where invaders will establish and where they will exert the strongest ecological impacts are key challenges for early detection and targeted management. Although Species Distribution Models (SDMs) are widely used to forecast biological invasions, they often provide uncertain estimates of establishment ranges and limited insight into invader performance, making it difficult to anticipate ecological impacts. Here, we address these limitations by integrating physiological information on invader performance with SDMs to identify regions of high invasion risk. Using the brown alga Rugulopteryx okamurae, one of the most prominent marine invaders in Europe, we first test alternative hypotheses of northern establishment limits: (i) a cold-survival constraint driven by winter temperatures and (ii) a growth constraint derived from the species' thermal performance. To identify the more likely scenario, we combine cold-tolerance experiments with seasonal growth comparisons between the invader and a native macroalga Dictyota dichotoma, whose established distribution allows physiological performance to be directly related to realised presence. Finally, we project seasonal growth of the invader across the predicted establishment range as a proxy for biomass accumulation and potential ecological impacts. Our results indicate that northern limit in Europe will be more likely constrained by winter survival rather than growth, extending the potential establishment range of Rugulopteryx to mid-Norway. In contrast, the highest impacts are likely to remain concentrated in southern Europe, where thermal conditions sustain high year-round growth. Overall, our approach illustrates how understanding the physiological response of invaders to their environment can improve the interpretation of SDM outputs and help identify areas at greatest risk of impact within their potential establishment range.

ecology↗

A spatial atlas of the seaweed CO2-fixation machinery reveals a unique Rubisco condensation mechanism

Seaweeds (macroalgae) are important primary producers that sustain food webs in coastal ecosystems. Most algae accelerate inorganic carbon assimilation by actively concentrating CO2 in a Rubisco-rich specialized organelle called the pyrenoid. However, the molecular composition of this pathway is unknown in seaweeds. Here, we investigated the intracellular localization of 160 proteins associated with CO2 acquisition in the green seaweed Ulva (Sea lettuce). We assign 68 proteins to different pyrenoid subdomains and identify a consensus Ulva Rubisco binding motif revealing the molecular logic of the Ulva pyrenoid. We reveal Seaweed Ulva Pyrenoid Assembly 1 (SUPA1) as the core pyrenoid assembly factor. We show that Rubisco condensation is driven by a unique mechanism: the helical folding of SUPA1 motifs upon Rubisco binding, combined with steric hindrance that halves the available Rubisco binding sites from eight to four. Our data gives an unprecedented sub-cellular spatial understanding on seaweed carbon fixation and provides insights into the evolution of this important pathway in the global carbon cycle.

cell biology↗

Chromosome-level genome assemblies of the red algae Porphyra dioica and Porphyra linearis

As one of the earliest-diverging multicellular eukaryotic lineages, the bladed Bangiales (Rhodophyta) possess a deep evolutionary history with a central role in the multi-billion-dollar global seaweed aquaculture industry. Although North Atlantic representatives are emerging candidates for regional mariculture, the scarcity of high-quality genomic resources for these taxa hinders both fundamental research and commercial optimization. To address this, we present the first chromosome-level genome assemblies for two native European species: Porphyra dioica (150.44 Mbp) and Porphyra linearis (95.22 Mbp). By integrating Oxford Nanopore Technologies (ONT) long-read sequencing with Hi-C proximity ligation, we generated highly contiguous nuclear genomes resolved into five chromosomes. Structural gene models were predicted through the BRAKER3 pipeline, identifying 12,548 and 10,382 protein-coding genes for P. dioica and P. linearis, respectively. Subsequent homology-based functional annotation characterized 57.4% and 59.8% of these predicted proteins. Supplemented by circularized organellar genomes, these reference genomes provide a critical framework for future research, enabling comparative studies of Atlantic-Pacific divergence and facilitating the development of selective breeding programs for sustainable European aquaculture.

genomics↗

Rare sex but a long life sustain seaweed populations at the warm edge of their range

AimThe life cycle of many organisms is all but stable across their distribution range. Most commonly, populations respond to environmental variation by shifting the timing of reproductive events (phenology). Or more profoundly, populations may (partly) shift their mode of reproduction from sexual to asexual. Life cycle variation can impact reproductive success, gene flow, genetic diversity, and, ultimately, the evolutionary trajectory of populations. Understanding the factors that influence life cycle variation is essential for grasping the biology and ecological roles of species. This study investigates the variation in life cycles and its effects on the genetic diversity of a brown seaweed, Dictyota, across its European range. LocationNorth-East Atlantic Ocean and the Mediterranean Sea. TaxonDictyota dichotoma (Hudson) J.V. Lamouroux (Phaeophyceae, Dictyotales) MethodsWe monitored phenology, fertility and lifespan in Atlantic and Mediterranean populations at its northern and southern boundaries, and used microsatellite markers to assess how these factors influence genetic and genotypic diversity. ResultsWe observed significant differences in phenology, reproductive strategies, and genetic diversity among northern and southern European populations. In Mediterranean populations, D. dichotoma exhibited sporophytic dominance with gametophytes being extremely rare, suggesting a shift towards asexual reproduction. In contrast, North-East Atlantic populations displayed more pronounced seasonal reproductive patterns with higher frequencies of gametophytes, indicating predominant sexual reproduction. However, genetic analysis showed lower allelic richness and unique alleles in northern populations, whereas southern populations were genetically more diverse, reflecting historical biogeographic processes. Clonal reproduction was more pronounced in the Mediterranean populations, influencing the spatial genetic substructure and contributing to lower genotypic diversity compared to Atlantic populations. Main conclusionsOur findings demonstrate that life cycle variation and phenology in D. dichotoma are closely tied to regional environmental conditions and have significant implications for population structure and genetic diversity. Results highlight how shifts in reproductive strategies contribute to the evolutionary and ecological dynamics of marine macroalgae across biogeographical gradients.

ecology↗

Consolidating Ulva functional genomics: Gene editing and new selection systems

The green seaweed Ulva compressa is a promising model for functional biology. In addition to historical research on growth and development, -omics data and molecular tools for stable transformation are available. However, more efficient tools are needed to study gene function. Here, we expand the molecular toolkit for Ulva. We screened the survival of Ulva and its mutualistic bacteria on 14 selective agents and establish that Blasticidin deaminases (BSD or bsr) can be used as selectable markers to generate stable transgenic lines. We show that Cas9 and Cas12a RNPs are suitable for targeted mutagenesis and can generate genomic deletions up to 20 kb using the marker gene ADENINE PHOSPHORIBOSYLTRANSFERASE (APT). We demonstrate that targeted insertion of a selectable marker via homology-directed repair or co-editing with APT is possible for non- marker genes. We evaluated 31 vector configurations and found that bicistronic fusion of Cas9 to a resistance marker or incorporation of introns in Cas9 led to the most mutants. We used this to generate mutants in three non-marker genes using a co-editing strategy. This expanded molecular toolkit now enables us to reliably make gain- and loss-of-function mutants, additional optimizations will be necessary to allow for vector-based multiplex genome editing in Ulva.

molecular biology↗

Expanding the toolkit for ploidy manipulation in Chlamydomonas reinhardtii

SummaryO_LIWhole genome duplications, widely observed in plant lineages, have significant evolutionary and ecological impacts. Yet, our current understanding of the direct implications of ploidy shifts on short- and long-term plant evolution remains fragmentary, necessitating further investigations across multiple ploidy levels. C_LIO_LIChlamydomonas reinhardtii, a haploid green alga, is a valuable model organism with profound potential to study the impact of ploidy increase on the longer-term in a laboratory environment. This is partly due to the ability to successfully increase the ploidy level. C_LIO_LIWe developed a strategy to engineer ploidy in Chlamydomonas reinhardtii using a collection of non-interfering antibiotic selectable markers. This approach allows to induce higher ploidy levels in Chlamydomonas reinhardtii and is applicable to field isolates, which expands beyond specific auxotroph laboratory strains and broadens the genetic diversity of parental haploid strains that can be crossed. We implement flow cytometry for precise measurement of the genome size of strains of different ploidy. C_LIO_LIWe demonstrate the creation of diploids, triploids and tetraploids by engineering North American field isolates, broadening the application of synthetic biology principles in Chlamydomonas reinhardtii. C_LIO_LIOur study greatly facilitates the application of Chlamydomonas reinhardtii to study polyploidy, both in fundamental and applied settings. C_LI

synthetic biology↗

Asymmetric genome merging leads to gene expression novelty through nucleo-cytoplasmic disruptions and transcriptomic shock in Chlamydomonas triploids

Genome merging is a common phenomenon in many organisms, causing a wide range of consequences on phenotype, adaptation, and gene expression, among other effects, yet its broader implications are not well understood. Two consequences of genome merging on gene expression remain poorly understood: dosage effects and evolution of expression. In this study, we employed Chlamydomonas reinhardtii as a model to investigate the effects of asymmetric genome merging by crossing a diploid with a haploid strain to create a novel triploid line. Five independent clonal lineages derived from this triploid line were evolved for 425 asexual generations in a laboratory natural selection (LNS) experiment. Utilizing fitness assays, qPCR, and RNA-Seq, we assessed the immediate consequences of genome merging and subsequent evolution over time. Our findings reveal substantial alterations in gene expression, protein homeostasis (proteostasis) and cytonuclear stoichiometry. Notably, gene expression exhibited expression level dominance and transgressivity (i.e., expression level higher or lower than either parent). Ongoing expression level dominance and a pattern of "functional dominance" from the haploid parent was observed, alongside remarkable stability in expression patterns across generations. Despite major nucleo-cytoplasmic disruptions, enhanced fitness was detected in the triploid strain. By comparing gene expression across generations, our results indicate that proteostasis restoration is a critical component of rapid adaptation following genome merging in Chlamydomonas reinhardtii and possibly other systems.

evolutionary biology↗

Breaking free from references: a consensus-based approach for community profiling with long amplicon nanopore data

Third-generation sequencing platforms, such as Oxford Nanopore Technology (ONT), have made it possible to characterise communities through the sequencing of long amplicons. Whilst this theoretically allows for an increased taxonomic resolution compared to short-read sequencing platforms such as Illumina, the high error rate remains problematic to accurately identify the community members present within a sample. Here we present and validate CONCOMPRA, a tool that allows the detection of closely related strains within a community by drafting and mapping to consensus sequences. We show that CONCOMPRA outperforms several other tools for profiling bacterial communities using full-length 16S rRNA gene sequencing. Since CONCOMPRA does not rely on a sequence database for profiling communities, it is applicable to systems and amplicons for which a reference framework is poorly developed. Our validation test shows that the amplification of long PCR products is likely to produce chimeric byproducts that inflate alpha diversity and skew community structure, stressing the importance of chimera detection. CONCOMPRA is available on GitHub (https://github.com/willem-stock/CONCOMPRA). Key points- We introduce a reference free tool for (long) amplicon Oxford Nanopore sequencing data - We show that it is capable of outperforming achieving a higher accuracy than preexisting tools - Chimera detection and removal is an essential step in the processing of long amplicon Oxford Nanopore sequencing data

bioinformatics↗

Functional stability despite high taxonomic turnover characterizes the Ulva microbiome across a 2,000 km salinity gradient

The green seaweed Ulva depends on its associated bacteria for morphogenesis and is an important model to study algal-bacterial interactions. Ulva-associated bacteria exhibit high turnover across environmental gradients, leading to the hypothesis that bacteria contribute to the acclimation potential of the host. Yet little is known about the variation in the functional profile of Ulva-associated bacteria in relation to environmental changes. To test which microbial functions shift alongside a strong environmental gradient, we analysed microbial communities of 91 Ulva samples across a 2,000 km Atlantic-Baltic Sea salinity gradient using metagenomic sequencing. Metabolic reconstruction of 639 metagenome-assembled genomes revealed widespread potential for carbon, sulphur, nitrogen, and vitamin metabolism, including amino acid and vitamin B biosynthesis. While salinity explained 70% of taxonomic variation, it only accounted for 17% of functional variation, indicating extensive functional stability. The limited variation was attributed to typical high-salinity bacteria exhibiting enrichment in genes for thiamine, pyridoxal, and betaine biosynthesis. These metabolic modules likely contribute to oxidative stress mitigation, cellular osmotic homeostasis, and membrane stabilization in response to salinity variations. Our results emphasise the importance of functional profiling to understand the seaweed holobiont and its collective response to environmental change.

microbiology↗

Cell wall-mediated maternal control of apical-basal patterning of the kelp Undaria pinnatifida

The role of maternal tissue in the control of embryogenesis remains enigmatic in many complex organisms. Here, we investigate the contribution of maternal tissue to apical-basal patterning in the kelp embryo. Using a modified kelp fertilisation protocol which yields synchronously developing kelp embryos, we show that detachment from maternal tissue leads to compromised robustness of apical-basal patterning. Detached embryos are rounder and often show aberrant morphologies. Furthermore, absence of contact with maternal tissue increases parthenogenesis, highlighting the critical role of maternal signals in the initial stages of kelp development. When zygotes are detached from the female gametophyte while part of the oogonial cell wall still remains attached to the egg, the proper apical-basal patterning is rescued showing a key role for the connection to the maternal cell wall in developmental patterning in kelps. This observation is reminiscent of another brown alga, Fucus, where the cell wall has been shown to play a key role in cell fate determination. In the case of kelps, the maternal oogonium mediates basal cell fate determination by providing an extrinsic patterning cue in its extracellular matrix to the future embryo. Our findings suggest a conserved mechanism across phylogenetically distant oogamous brown algal lineages, where localised secretion of sulphated F2 fucans mediate establishment of the apical-basal polarity.

developmental biology↗

Gametogenesis in the green seaweed Ulva mutabilis coincides with massive transcriptional restructuring

The molecular mechanism underlying sexual reproduction in land plants is well understood in model plants and is a target for crop improvement. However, unlike land plants, the genetic basis involved in triggering reproduction and gamete formation remains elusive in most seaweeds, which are increasingly viewed as an alternative source of functional food and feedstock for energy applications. Here, gametogenesis of Ulva mutabilis, a model organism for green seaweeds, is studied. We analyze transcriptome dynamics at different time points during gametogenesis following induction of reproduction by fragmentation and removal of sporulation inhibitors. Analyses demonstrate that 45% of the genes in the genome are differentially expressed during gametogenesis. We identified several transcription factors that potentially play a key role in the early gametogenesis of Ulva given the function of their homologs in higher plants and microalgae. In particular, the detailed expression pattern of an evolutionary conserved transcription factor containing an RWP-RK domain suggests a key role during Ulva gametogenesis. The identification of putative master regulators of gametogenesis provides a starting point for further functional characterization. HighlightTranscriptomic analyses of gametogenesis in the green seaweed Ulva highlight the importance of a conserved RWP-RK transcription factor in induction of sexual reproduction.

plant biology↗

A molecular toolkit for the green seaweed Ulva mutabilis

The green seaweed Ulva is an ecologically-important marine primary producer as well as a promising cash crop cultivated for multiple uses. Despite its importance, several molecular tools are still needed to better understand seaweed biology. Here, we report the development of a flexible and modular molecular cloning toolkit for the green seaweed Ulva mutabilis based on a Golden Gate cloning system. The toolkit presently contains 125 entry vectors, 26 destination vectors and 107 functionally validated expression vectors. We demonstrate the importance of endogenous regulatory sequences for transgene expression and characterize three endogenous promoters suitable to drive transgene expression. We describe two vector architectures to express transgenes via two expression cassettes or a bicistronic approach. The majority of selected transformants (50-80%) consistently give clear visual transgene expression. Furthermore, we made different marker lines for intracellular compartments after evaluating 13 transit peptides and 11 tagged endogenous Ulva genes. Our molecular toolkit enables the study of Ulva gain-of-function lines and paves the way for gene characterization and large-scale functional genomics studies in a green seaweed. One-sentence summaryMolecular cloning tools allow to generate gain-of-function seaweed lines that will help to study seaweed biology.

plant biology↗

Neoproterozoic origin and multiple transitions to macroscopic growth in green seaweeds

The Neoproterozoic Era records the transition from a largely bacterial to a predominantly eukaryotic phototrophic world, creating the foundation for the complex benthic ecosystems that have sustained Metazoa from the Ediacaran Period onward. This study focusses on the evolutionary origins of green seaweeds, which play an important ecological role in the benthos of modern sunlit oceans and likely played a crucial part in the evolution of early animals by structuring benthic habitats and providing novel niches. By applying a phylogenomic approach, we resolve deep relationships of the core Chlorophyta (Ulvophyceae or green seaweeds, and freshwater or terrestrial Chlorophyceae and Trebouxiophyceae) and unveil a rapid radiation of Chlorophyceae and the principal lineages of the Ulvophyceae late in the Neoproterozoic Era. Our time-calibrated tree points to an origin and early diversification of green seaweeds in the late Tonian and Cryogenian periods, an interval marked by two global glaciations, with strong consequent changes in the amount of available marine benthic habitat. We hypothesize that the unicellular and simple multicellular ancestors of green seaweeds survived these extreme climate events in isolated refugia, and diversified following recolonization of benthic environments that became increasingly available as sea ice retreated. An increased supply of nutrients and biotic interactions such as grazing pressure has likely triggered the independent evolution of macroscopic growth via different strategies, including both true multicellularity, and multiple types of giant celled forms.\n\nSignificance StatementGreen seaweeds are important primary producers along coastlines worldwide, and likely played a key role in the evolution of animals. To understand their origin and diversification, we resolve key relationships among extant green algae using a phylotranscriptomic approach. We calibrate our tree using available fossil data, to reconstruct important evolutionary events such as transitions to benthic environments, and evolution of macroscopic growth. We estimate green seaweeds to have originated in the late Tonian/Cryogenian Period, followed by a marked Ordovician diversification of macroscopic forms. This ancient proliferation of green seaweeds likely modified shallow marine ecosystems, which set off an evolutionary arms race between ever larger seaweeds and grazers.

evolutionary biology↗

Population structure and geographically structured reproductive strategies of the haplodiplontic seaweed Dictyota dichotoma

Both mating system variation and the propensity of many seaweeds to reproduce both sexually and asexually, leave a strong imprint in the genetic structure of species. In this respect, we study the population genetic structure of Dictyota dichotoma, a common haplodiplont brown subtidal seaweed. This benthic species is widespread in the NE-Atlantic, from the Canary Islands and Mediterranean Sea to southern Norway, but lately populations have been reported from Argentina and South Africa. Phenology and reproduction of D. dichotoma was monitored year-round in four populations to investigate how the species has adapted to the steep thermal gradient in southern and northern ranges of its distribution. Thirteen microsatellites are developed in order to assess patterns of population diversity and structure across the biogeographic range, as shaped by past and present processes. Last, we assess the genetic structure of South African and South American populations and their relationship to the northern hemisphere populations.\n\nThroughout its range, D. dichotoma shows a varying reproductive effort, with sexual reproduction being more abundant in the northern range. In contrast, the Mediterranean populations show a clear sporophyte dominance, suggesting that sexual reproduction is not the prime mode of reproduction, and indicating that the species potentially resorts to other modes of propagation as for instance fragmentation or apospory.\n\nGenetic diversity is highest in the southern population decreasing gradually northward, indicative for a recolonization pattern after the demise of the last glacial maximum where these areas served as glacial refugia. European mainland populations show an isolation by distance pattern, while the population in the Canary Islands has its own genetic identity, being significantly diverged from the mainland population. Populations in South Africa and Argentina are seemingly introduced from mainland Europe, but no conclusion can be made on the exact timing of these introductions.

ecology↗

The Plastid Genome In Cladophorales Green Algae Is Encoded By Hairpin Plasmids

Virtually all plastid (chloroplast) genomes are circular double-stranded DNA molecules, typically between 100-200 kb in size and encoding circa 80-250 genes. Exceptions to this universal plastid genome architecture are very few and include the dinoflagellates where genes are located on DNA minicircles. Here we report on the highly deviant chloroplast genome of Cladophorales green algae, which is entirely fragmented into hairpin plasmids. Short and long read high-throughput sequencing of DNA and RNA demonstrated that the chloroplast genes of Boodlea composita are encoded on 1-7 kb DNA contigs with an exceptionally high GC-content, each containing a long inverted repeat with one or two protein-coding genes and conserved non-coding regions putatively involved in replication and/or expression. We propose that these contigs correspond to linear single-stranded DNA molecules that fold onto themselves to form hairpin plasmids. The Boodlea chloroplast genes are highly divergent from their corresponding orthologs. The origin of this highly deviant chloroplast genome likely occurred before the emergence of the Cladophorales, and coincided with an elevated transfer of chloroplast genes to the nucleus. A chloroplast genome that is composed only of linear DNA molecules is unprecedented among eukaryotes and highlights unexpected variation in the plastid genome architecture.

genomics↗