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Gouvea, L.

Publications and source records attributed to Gouvea, L..

2 recordsLinked to original sources

SAMMBA is a high-throughput pipeline for isolating and phenotyping macroalgal strains

Anthropogenic climate change is causing the decline of seaweed forests in many parts of the world. Despite successful preservation efforts, their immense biodiversity is still severely underrepresented in germplasm biobanks throughout the world. These culture libraries can preserve genetic diversity and provide inoculum for marine forest restoration and mariculture ventures, and potentially accelerate the selection and breeding of climate-resilient and high-yielding strains. However, the complex life cycles and body plans of seaweeds pose a huge challenge for the development of standardized phenotyping and isolating protocols for microscopic stages, especially with the efficiency necessary to deal with the current pace of global climatic changes. 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 cells in 384-well plates (384WP). By optimizing fluorescence microscopy imaging and analysis, along with a novel fragmentation method and dilution-to-extinction isolation, different unialgal seaweed tissues could be regrown after thousand-fold dilutions. In a single plate, we successfully isolated 68 singlet gametophyte fragments of Laminaria ochroleuca (39 males, 29 females; 17.7% efficiency) and 60 spores of Phyllariopsis purpurascens (31.25% efficiency). Furthermore, the taxonomic versatility of SAMMBA was demonstrated through the successful isolation of 60 unialgal cultures of red algae (Halymenia sp., Hydrolithon sp., Erythrotrichia sp.) and 10 strains of the green alga Ulva sp, without cross-contamination. The viability and unialgal nature of the isolated strains were verified by distributing a single L. ochroleuca strain across an entire 384-well plate and imaging each well over 30 days. We found that the average specific daily growth rates (daily SGR) per well were 0.130 {+/-} 0.006 and 0.117 {+/-} 0.01 day-1 for males and females, respectively, showing a significant difference between sexes (n = 768; p = 1.27e-53), while edge effects significantly reduced daily SGR in males but not in females. This approach dramatically increases experimental reproducibility and statistical power compared to conventional methods. Due to its modular design and cost-effectiveness, SAMMBA is readily adaptable to macroalgal repositories globally. It supports high-throughput, selective recovery of unialgal strains without reliance on robotic platforms, while remaining fully compatible with automation. This system significantly expands the experimental and operational capacity in macroalgal hatcheries, providing a scalable foundation for phenomics, domestication programs, and standardized, verifiable biobanking of unialgal strains. Ultimately, SAMMBA could provide critical support for breeding strategies required to ensure the resilience of marine forests and aquaculture in a rapidly changing ocean.

plant biology↗

Unravelling the role of oceanographic connectivity in intra-specific diversity of marine forests at global scale

AimIntra-specific diversity results from complex interactions of intermingled eco-evolutionary processes along species history, but their relative contribution has not been addressed at the global scale. Here, we unravel the role of present-day oceanographic connectivity in explaining the genetic differentiation of marine forests across the ocean. LocationGlobal. Time periodContemporary. Major taxa studiedMarine forests of brown macroalgae (order Fucales, Ishigeales, Laminariales, Tilopteridale). MethodsThrough systematic literature revision, we compiled a comprehensive dataset of genetic differentiation, encompassing 662 populations of 34 species. A biophysical model coupled with network analyses estimated multigenerational oceanographic connectivity and centrality across the marine forest global distribution. This approach integrated species dispersive capacity and long-distance dispersal events. Linear mixed models tested the relative contribution of site-specific processes, connectivity, and centrality in explaining genetic differentiation. ResultsWe show that spatiality dependent eco-evolutionary processes, as described by our models, are prominent drivers of genetic differentiation in marine forests (significant models in 92.6 % of the cases with an average R2 of 0.49 {+/-} 0.07). Specifically, we reveal that 19.6 % of variance is explicitly induced by contemporary connectivity and centrality. Moreover, we demonstrate that LDD is key in connecting populations of species distributed across large water masses and continents. Main conclusionsWe deciphered the role of present-day connectivity in observed patterns of genetic differentiation of marine forests. Our findings significantly contribute to the understanding of the drivers of intra-specific diversity on a global scale, with implications for biogeography and evolution. These results can guide well-informed conservation efforts, including the designation of marine protected areas, as well as spatial planning for genetic diversity in aquaculture, which is particularly relevant for sessile ecosystems structuring species such as brown macroalgae.

ecology↗