Bull kelp, Nereocystis luetkeana, is a northeastern Pacific kelp with a broad distribution from Alaska to central California. Its population declines have caused severe concerns in northern California, the Salish Sea in Washington, and recently in some populations in Oregon. Despite bull kelp's accumulated ecological and physiological studies, an assembled and annotated genomic reference was still unavailable. Here, we report the complete and annotated genome of N. luetkeana, produced by the California Conservation Genomics Project (CCGP), which aims to reveal genomic diversity patterns across California by sequencing the complete genomes of approximately 150 carefully selected species. The genome was assembled into 1,562 scaffolds with 449.82 Mb, 80× of coverage, and 22,952 gene models. BUSCO assembly showed a completeness score of 72% for the stramenopiles gene set. The mitochondria and chloroplast genome sequences have 37 Kb and 131 Mb, respectively. The orthology analysis between 10 Phaeophycean genomes showed 1,065 expanded and 286 unique orthogroups for this species. Pairwise comparisons showed 542 orthogroups present only in N. luetkeana and Macrocystis pyrifera, another large-body kelp. The enrichment analysis of these orthogroups showed important functions related to central metabolism and signaling due to ATPase enrichment in these two species. This genome assembly will provide an essential resource for the ecology, evolution, conservation, and breeding of bull kelp.

Seaweed farming, as a part of aquaculture, offers a sustainable alternative to modern agricultural practices; however, genetic enhancement and breeding programs for most species are underdeveloped. We aimed to advance seaweed domestication by focusing on giant kelp (Macrocystis pyrifera), the fastest-growing haplodiplontic brown alga, which has significant ecological and commercial importance. We analyzed phenotypic data from two offshore experimental farms conducted in 2019 and 2020, which involved hundreds of outplanted genetically diverse sporophytes. We found that outplanting season and farm design had significant effects on giant kelp biomass. Broad-sense heritability estimates showed moderate (0.27-0.50) genetic contributions to two phenotypes, carbon content and total biomass. Genome-wide association studies for these phenotypes resulted in three statistically significant SNPs, located near or within genes involved in carbohydrate metabolism and cytoskeletal functions. In addition, we applied genomic selection models that integrated sporophyte phenotypes and parental gametophyte genotypes. These models utilized reduced sets of GWAS-ranked SNPs obtained by a procedure based on linkage disequilibrium estimations. Model testing yielded cross-validation accuracy values of up to 0.84 and predictive accuracy values of up to 0.40, demonstrating the potential of marker-assisted breeding for phenotype improvement. Our results provide foundational genomic resources and tools for domesticating and breeding M. pyrifera, forming a basis for developing giant kelp varieties with desirable traits.

Despite successful preservation efforts, macroalgal diversity remains under-represented in global biobanks. A major limitation is the extreme morphological diversity of seaweed thalli, which hinders standardized isolation and phenotyping and often requires taxon-specific protocols. Here we present SAMMBA (Seaweed Automatable Microplate Microscopy for Breeding Approaches), an end-to-end pipeline for the high-throughput isolation, phenotyping and storage of macroalgal propagules in 384-well plates. By optimizing live-cell manipulation for chlorophyll autofluorescence (CAF) imaging and segmentation, multiple unialgal propagules can be isolated by dilution-based workflows. In a single plate, we obtained 68 singlet gametophyte fragments of Laminaria ochroleuca (17.7%) and 60 meiospores of Phyllariopsis purpurascens (31.25%). We demonstrated taxonomic and morphological versatility by isolating 60 unialgal cultures from three distinct Rhodophyta morphotypes (filamentous, crustose and foliose) and 10 strains of Ulva sp., also in a single plate. Furthermore, CAF-based area increase over 30 days enabled high-precision estimates of specific growth rates, yielding 0.130 +/- 0.006 and 0.117 +/- 0.01 day- 1 for male and female L. ochroleuca gametophytes, respectively (n = 768; p = 1.27e-53). This precision substantially increases experimental reproducibility and statistical power compared to conventional methods, supporting high-throughput recovery of unialgal strains without motorized platforms, while remaining fully compatible with automation. SAMMBA expands operational capacity for strain discovery and phenotyping, providing a scalable foundation for phenomics, domestication workflows, and standardized macroalgal biobanking. We outline how the platform can benefit multiple areas of phycological research and facilitate the development of improved strains that can support aquaculture and restoration efforts.

Kelp, brown macroalgae in the order Laminariales, provide ecosystem services vital to ocean biodiversity. However, kelp forests worldwide are declining due to abiotic stressors such as ocean warming. In this study, we present results from high-resolution confocal microscopy and in vivo imaging system imaging using protocols developed to visualize kelp gametophyte cells exposed to heat-stress treatments. Imaging revealed chloroplast mislocalization, fragmentation, and subsequent loss of chloroplasts in heat-stressed gametophyte cells. Additionally, nuclei exhibited fragmentation and a progressive loss of fluorescent signal, and the associated microbiome proliferated under various heat-stress treatments. Notably, because brown algae possess a continuous outer membrane that connects the nuclear envelope and the chloroplast envelope, these observations suggest a cellular vulnerability underlying thermal sensitivity in brown macroalgae. Finally, by comparing heat-stress tolerant and heat-stress sensitive genotypes, we found that genotypes with higher heat tolerance exhibited substantially fewer abnormalities compared to sensitive ones.

Stark declines in the abundance and distribution of bull kelp and reduced genetic diversity have been observed in Puget Sound, Washington, especially in southernmost areas. Consequently, introducing new variation through genetic rescue (GR) has emerged as a promising restoration strategy. However, GR success hinges on the preservation of local adaptation, which has not yet been studied in the region. To that end, we performed whole-genome sequencing on 100 gametophyte individuals from our biobank, sourced from 14 populations across Puget Sound. We identified three genetic clusters, corresponding broadly to southern Puget Sound (SPS), Whidbey Basin (WB), and the Strait of Juan de Fuca (SJF). Despite low genetic diversity and high genetic load, we found evidence of local adaptation to five environmental variables. Although salinity emerged as the strongest predictor of environmentally associated genetic variance overall, in SPS the primary drivers were average winter sea surface temperature (SST) and seasonal turbidity. We identified SNPs and enriched gene ontology (GO) terms and analyzed their allele frequencies to assess functional variation between clusters. These frequencies revealed marked differences between SJF and the other clusters, while for most genes, the highest-frequency allele in WB was also most prevalent in SPS. However, for all genes linked to winter SST and turbidity, SPS harbored distinct alleles not found in any other cluster. These findings suggest that a WB population would likely be a strong candidate donor for SPS due to its higher genetic diversity and shared adaptive variation. Further experiments are needed to validate these results and assess the suitability of WB populations for GR. ### Competing Interest Statement The authors have declared no competing interest. Washington Sea Grant, https://ror.org/047fr0y29, UWSC13569

Microbial inoculants can increase the yield of cultivated crops and are successful in independent trials; however, efficacy drops in large-scale applications due to insufficient consideration of microbial community dynamics. The structure of microbiomes, in addition to the impact of individual taxa, is an important factor to consider when designing growth-promoting inoculants. Here, we investigate the microbial network and community assembly patterns of Macrocystis pyrifera gametophyte germplasm cultures (collectively referred to as a “seedbank”) used to cultivate an offshore farm in Santa Barbara, California, and identify network features associated with increased biomass of mature sporophytes. We found that [1] several network features, such as clustering coefficient and edge ratios, significantly vary with biomass outcomes; [2] gametophytes that become low- or high-biomass sporophytes have different hub taxa; and [3] microbial community assembly of gametophyte germplasm cultures is niche-driven. Overall, this study describes microbial community dynamics in M. pyrifera germplasm cultures and ultimately supports the development of early life stage inoculants that can be used on seaweed cultivars to increase biomass yield.

Kelps are vital for marine ecosystems, yet the genetic diversity underlying their capacity to adapt to climate change remains unknown. In this study, we focused on the kelp Macrocystis pyrifera a species critical to coastal habitats. We developed a protocol to evaluate heat stress response in 204 Macrocystis pyrifera genotypes subjected to heat stress treatments ranging from 21 degrees C to 27 degrees C. Here we show that haploid gametophytes exhibiting a heat-stress tolerant (HST) phenotype also produced greater biomass as genetically similar diploid sporophytes in a warm-water ocean farm. HST was measured as chlorophyll autofluorescence per genotype, presented here as fluorescent intensity values. This correlation suggests a predictive relationship between the growth performance of the early microscopic gametophyte stage HST and the later macroscopic sporophyte stage, indicating the potential for selecting resilient kelp strains under warmer ocean temperatures. However, HST kelps showed reduced genetic variation, underscoring the importance of integrating heat tolerance genes into a broader genetic pool to maintain the adaptability of kelp populations in the face of climate change.

Abstract Kelps are vital for marine ecosystems, yet the genetic diversity underlying their capacity to adapt to climate change remains unknown. In this study, we focused on the kelp Macrocystis pyrifera (M. pyrifera), a species critical to coastal habitats. We developed a phenotyping protocol to evaluate heat stress response in 204 M. pyrifera gametophytes, subjected to heat stress treatments ranging from 21°C to 27°C. Here we show that genotypes exhibiting a heat-stress tolerant (HST) phenotype as haploid gametophytes also produced greater biomass as diploid sporophytes in a warm-water ocean farm. This correlation suggests a predictive relationship between early-stage HST and later-stage growth performance, indicating the potential for selecting resilient kelp strains under warmer ocean temperatures. However, HST kelps showed reduced genetic variation, underscoring the importance of integrating heat tolerance genes into a broader genetic pool to maintain the adaptability of kelp populations in the face of climate change.

Macrocystis pyrifera (giant kelp), is a brown macroalga of great ecological importance as a primary producer and structure-forming foundational species that provides habitat for hundreds of species. It has many commercial uses (e.g. source of alginate, fertilizer, cosmetics, feedstock). One of the limitations to exploiting giant kelp’s economic potential and assisting in giant kelp conservation efforts is a lack of genomic tools like a high quality, contiguous reference genome with accurate gene annotations. Reference genomes attempt to capture the complete genomic sequence of an individual or species, and importantly provide a universal structure for comparison across a multitude of genetic experiments, both within and between species. We assembled the giant kelp genome of a haploid female gametophyte de novo using PacBio reads, then ordered contigs into chromosome level scaffolds using Hi-C. We found the giant kelp genome to be 537 MB, with a total of 35 scaffolds and 188 contigs. The assembly N50 is 13,669,674 with GC content of 50.37%. We assessed the genome completeness using BUSCO, and found giant kelp contained 94% of the BUSCO genes from the stramenopile clade. Annotation of the giant kelp genome revealed 25,919 genes. Additionally, we present genetic variation data based on 48 diploid giant kelp sporophytes from three different Southern California populations that confirms the population structure found in other studies of these populations. This work resulted in a high-quality giant kelp genome that greatly increases the genetic knowledge of this ecologically and economically vital species.

IntroductionIn temperate regions, one of the most critical determinants of present range-wide genetic diversity was the Pleistocene climate oscillations, the most recent one created by the last glacial maximum (LGM). This study aimed to describe N. luetkeana genetic structure across its entire range (Alaska to California) and test different models of population connectivity within the Salish Sea. This region was colonized after the LGM and has been under increased disturbance in recent decades.MethodsWe utilized microsatellite markers to study N. luetkeana genetic diversity at 53 sites across its range. Using higher sampling density in the Salish Sea, we employed a seascape genetics approach and tested isolation by hydrodynamic transport and environment models.ResultsAt the species distribution scale, we found four main groups of genetic co-ancestry, Alaska; Washington with Vancouver Island’s outer coast and Juan de Fuca Strait; Washington’s inner Salish Sea; and Oregon with California. The highest allelic richness (AR) levels were found in California, near the trailing range edge, although AR was also high in Alaska. The inner Salish Sea region had the poorest diversity across the species distribution. Nevertheless, a pattern of isolation by hydrodynamic transport and environment was supported in this region.DiscussionThe levels of allelic, private allele richness and genetic differentiation suggest that during the LGM, bull kelp had both northern and southern glacial refugia in the Prince of Wales Island-Haida Gwaii region and Central California, respectively. Genetic diversity in Northern California sites seems resilient to recent disturbances, whereas the low levels of genetic diversity in the inner Salish Sea are concerning.

Giant kelp, Macrocystis pyrifera, exists as distinct morphological variants-or "ecomorphs"-in different populations, yet the mechanism for this variation is uncertain, and environmental drivers for either adaptive or plastic phenotypes have not been identified. The ecomorphs Macrocystis "pyrifera" and M. "integrifolia" are distributed throughout temperate waters of North and South America with almost no geographic overlap and exhibit an incongruous, non-mirrored, distribution across the equator. This study evaluates the degree of genetic divergence between M. "pyrifera" and M. "integrifolia" across 18 populations in Chile and California using whole-genome sequencing and single-nucleotide polymorphism markers. Our results based on a principal component analysis, admixture clustering by genetic similarity, and phylogenetic inference demonstrate that M. "pyrifera" and M. "integrifolia" are genetically distinguishable. Analyses reveal separation by Northern and Southern Hemispheres and between morphs within hemispheres, suggesting that the convergent "integrifolia" morphology arose separately in each hemisphere. This is the first study to use whole-genome sequencing to understand genetic divergence in giant kelp ecomorphs, identifying 83 potential genes under selection and providing novel insights about Macrocystis evolution that were not evident with previous genetic techniques. Future studies are needed to uncover the environmental forces driving local adaptation and presumed convergent evolution of these morphs.

With national interest in seaweed-based biofuels as a sustainable alternative to fossil fuels, there is a need for tools that produce high-yield seaweed cultivars and increase the efficiency of offshore farms. Several agricultural studies have demonstrated that the application of microbial inoculants at an early life stage can improve crop yield, and there is an opportunity to use similar techniques in seaweed aquaculture. However, there is a critical knowledge gap regarding host-microbiome associations of macroalgae gametophytes in germplasm cultures. Here, we investigate the microbial community of Macrocystis pyrifera gametophyte germplasm cultures that were used to cultivate an offshore farm in Santa Barbara, California and identify key taxa correlated with increased biomass of mature sporophytes. This work provides a valuable knowledge base for the development of microbial inoculants that produce high-biomass M. pyrifera cultivars to ultimately be used as biofuel feedstocks.

Kelp species provide many ecosystem services associated with their three-dimensional structures. Among these, fast-growth, canopy-forming species, like giant kelp Macrocystis pyrifera, are the foundation of kelp forests across many temperate reefs. Giant kelp populations have experienced regional declines in different parts of the world. Giant kelp canopy is very dynamic and can take years to recover from disturbance, challenging comparisons of standing biomass with historical baselines. The Santa Barbara Coastal LTER (SBC LTER), curates a time series of Landsat sensed surface cover and biomass for giant kelp in the west coast of North America. In the last decade, this resource has been fundamental to understanding the species' population dynamics and drivers. However, simple ready-to-use summary statistics aimed at classifying regional kelp decline or recovery are not readily available to stakeholders and coastal managers. To this end, we describe here two simple metrics made available through the R package kelpdecline. First, the proportion of Landsat pixels in decline (PPD), in which current biomass is compared with a historical baseline, and second, a pixel occupancy trend (POT), in which current year pixel occupancy is compared to the time-series long probability of occupancy. The package produces raster maps and output tables summarizing kelp decline and trends over a 0.25 × 0.25° scale. Using kelpdecline, we show how sensitivity analysis on PPD parameter variation can increase the confidence of kelp decline estimates.

The paradigm of past climate-driven range shifts structuring the distribution of marine intraspecific biodiversity lacks replication in biological models exposed to comparable limiting conditions in independent regions. This may lead to confounding effects unlinked to climate drivers. We aim to fill in this gap by asking whether the global distribution of intraspecific biodiversity of giant kelp (Macrocystis pyrifera) is explained by past climate changes occurring across the two hemispheres. We compared the species’ population genetic diversity and structure inferred with microsatellite markers, with range shifts and long-term refugial regions predicted with species distribution modelling (SDM) from the last glacial maximum (LGM) to the present. The broad antitropical distribution of Macrocystis pyrifera is composed by six significantly differentiated genetic groups, for which current genetic diversity levels match the expectations of past climate changes. Range shifts from the LGM to the present structured low latitude refugial regions where genetic relics with higher and unique diversity were found (particularly in the Channel Islands of California and in Peru), while post-glacial expansions following ~ 40% range contraction explained extensive regions with homogenous reduced diversity. The estimated effect of past climate-driven range shifts was comparable between hemispheres, largely demonstrating that the distribution of intraspecific marine biodiversity can be structured by comparable evolutionary forces across the global ocean. Additionally, the differentiation and endemicity of regional genetic groups, confers high conservation value to these localized intraspecific biodiversity hotspots of giant kelp forests.

Macrocystis pyrifera (giant kelp), a haplodiplontic brown macroalga that alternates between a macroscopic diploid (sporophyte) and a microscopic haploid (gametophyte) phase, provides an ideal system to investigate how ploidy background affects the evolutionary history of a gene. In M. pyrifera , the same genome is subjected to different selective pressures and environments as it alternates between haploid and diploid life stages. We assembled M. pyrifera gene models using available expression data and validated 8,292 genes models using the model alga Ectocarpus siliculosus . Differential expression analysis identified gene models expressed in either or both the haploid and diploid life stages while functional annotation identified processes enriched in each stage. Genes expressed preferentially or exclusively in the gametophyte stage were found to have higher nucleotide diversity (π = 2.3 × 10 –3 and 2.8 × 10 –3 , respectively) than those for sporophytes (π = 1.1 × 10 –3 and 1 × 10 –3 , respectively). While gametophyte-biased genes show faster sequence evolution, the sequence evolution exhibits less signatures of adaptations when compared to sporophyte-biased genes. Our findings contrast the standing masking hypothesis, which predicts higher standing genetic variation at the sporophyte stage, and support the strength of expression theory, which posits that genes expressed more strongly are expected to evolve slower. We argue that the sporophyte stage undergoes more stringent selection compared with the gametophyte stage, which carries a heavy genetic load associated with broadcast spawning. Furthermore, using whole-genome sequencing, we confirm the strong population structure in wild M. pyrifera populations previously established using microsatellite markers, and estimate population genetic parameters, such as pairwise genetic diversity and Tajima’s D , important for conservation and domestication of M. pyrifera .

Given the impacts of climate change and other anthropogenic stressors on marine systems, there is a need to accurately predict how species respond to changing environments and disturbance regimes. The use of genetic tools to monitor temporal trends in populations gives ecologists the ability to estimate changes in genetic diversity and effective population size that may be undetectable by traditional census methods. Although multiple studies have used temporal genetic analysis, they usually involve commercially important species, and rarely sample before and after disturbance. In this study, we run a temporal analysis of giant kelp, Macrocystis pyrifera, genetic diversity over the scope of 10 years (2008-2018) using the same microsatellite marker panel to assess the genetic consequences of disturbance in several populations of giant kelp (Macrocystis pyrifera) in the Southern California Bight. The study is a rare pre- and post-disturbance microsatellite analysis that included declines to giant kelp caused by the 2015/16 El Nino Southern Oscillation event. We used canopy biomass estimated by remote sensing (Landsat) to quantify the extent of disturbance to kelp beds, and sea surface temperature data to understand how kelp was pushed towards its temperature limits during this period. Despite prolonged periods with decreased canopy at several sites, no changes in genetic structure and allelic richness were observed. We argue that giant kelp in the region is best described as a “patchy population” system where true extinctions are rare. We discuss how deep refugia of subsurface sporophytes and cryptic microscopic life stages could have kept genetic diversity through disturbance. Given the increasing effects of climate change and uncertainty in modeling impacts of species with cryptic life history stages, we suggest further investigation to reveal the role such stages play in species resilience. Genetic monitoring studies of sites selected by remote census demographic and climate surveys should be continued in the future given the predicted impacts of climate change.

Seagrass meadows are crucial habitats on nearshore areas, which are exposed to human disturbances that frequently cause seagrass loss. Although demographic and mapping data have been widely used in long-term monitoring programmes, the integration of multi-locus genotype data through time remains rare. The present work links, for the first time, landscape, demographic, and genetic data for a seagrass, so as to explore long-term responses to persistent human impacts. The temporal patterns in meadow area, shoot density, and clonal and genetic diversity of three Cymodocea nodosa meadows were compared: in one meadow directly affected by the construction of an industrial port and in two control areas. The hypothesis tested was that seagrass deterioration and subsequent habitat loss at the affected meadow would be associated with a reduction of seagrass clonal and genetic diversity. The results show significant reductions in both meadow area (from approx. 21 to 1.5 ha) and shoot density (from approx. 1,800 to 400 shoots per m(2)) at the impacted meadow, with concurrently reduced clonal and allelic richness (by approx. 22%), as well as heterozygosity (HLmean, approx. 21%; H-obs, approx. 12%). These descriptors, however, remained stable, or even increased, in the controls. Importantly for seagrass management, multi-disciplinary analysis made it possible to link slower, large-scale, landscape and genetic degradation processes with faster demographic deterioration on an intermediate meadow scale. Multi-locus genotype data were also crucial for revealing long-term degradation processes at the genetic level, which is undetectable by traditional monitoring techniques. In summary, this holistic approach provides a valuable framework for long-term seagrass monitoring programmes to detect cumulative signals of seagrass meadow degradation across a range of scales.

Ex situ seed banking was first conceptualized and implemented in the early 20th century to maintain and protect crop lines. Today, ex situ seed banking is important for the preservation of heirloom strains, biodiversity conservation and ecosystem restoration, and diverse research applications. However, these efforts primarily target microalgae and terrestrial plants. Although some collections include macroalgae (i.e., seaweeds), they are relatively few and have yet to be connected via any international, coordinated initiative. In this piece, we provide a brief introduction to macroalgal germplasm banking and its application to conservation, industry, and mariculture. We argue that concerted effort should be made globally in germline preservation of marine algal species via germplasm banking with an overview of the technical advances for feasibility and ensured success.