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High-throughput assessment of gametophyte survival and growth in the kelp Laminaria ochroleuca through autofluorescence analysis

2026-08-17Artigo científico Acesso aberto
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datacite.subject.sdg14:Proteger a Vida Marinha
datacite.subject.sdg15:Proteger a Vida Terrestre
datacite.subject.sdg09:Indústria, Inovação e Infraestruturas
dc.contributor.authorFélix de Azeredo Pinto e Melo, Francisca
dc.contributor.authorAlves-Lima, Cicero
dc.contributor.authorde Matos Barreto, Luís António
dc.contributor.authorMonico, Carina
dc.contributor.authorSerrao, Ester A.
dc.contributor.authorPearson, Gareth Anthony
dc.contributor.authorMartins, Neusa
dc.date.accessioned2026-08-26T12:11:59Z
dc.date.available2026-08-26T12:11:59Z
dc.date.issued2026-08-17
dc.description.abstractThe viability of kelp microscopic gametophytes is currently analyzed using subjective visual methods based on bright-field (BF) microscope images. Fluorescence microscopy (FM) can be employed, but the dyes used can be toxic and blue/UV light intensity may induce gametogenesis. This study aimed to develop a non-invasive and accurate methodology for assessing gametophyte viability over time, using FM observation of chlorophyll autofluorescence emitted by live gametophytes. Six isolated gametophyte strains of Laminaria ochroleuca (Italy), maintained at CCMAR Biobank, were cultured in triplicate (female, male, and both sexes combined) in Petri dishes containing 10 mL of half-strength Provasoli’s enriched seawater medium (PES), under red light for 1 month. Survival and growth were assessed on days 1, 7 and 14 by photographing 20 fields-of-view, both in BF and FM. Then, cultures were transferred to white light to induce gametogenesis and test whether the autofluorescence analysis (AFA) method affected gametophyte reproduction. Image analysis was performed using FIJI software either by manually counting live gametophytes (survival) and measuring their area (growth), or by implementing a machine-learning (ML) model using FIJI’s WEKA segmentation plugin. Results revealed that both methods were correlated, validating our ML model. However, the AFA method was faster and more accurate than BF image analysis, especially for male gametophytes, and did not compromise reproductive capacity. After 9 days under white light, sporophytes were present in mixed cultures. The AFA method provides a reliable technique for assessing gametophyte viability without compromising gametogenesis.eng
dc.description.sponsorshipUID/04326/2025; UID/PRR/04326/2025; MPr-2023–12-16551, ALGARVE-FEDER-00772500), POCI/MAR/60883/2004; PTDC/MAR/105152/2008, MPr-2023–12-16551 (ALGARVEFEDER-00772500) and from the operational programmes CRESC Algarve 2020 and COMPETE 2020 through contract EMBRC.PT ALG-01–0145-FEDER-022121. NM was also supported by MARPLUS—Support for the development of CCMAR’s European scientific infrastructure (ALGARVE-FSE+-01177700).
dc.identifier.doi10.1007/s10811-026-03989-4
dc.identifier.eissn1573-5176
dc.identifier.issn0921-8971
dc.identifier.urihttp://hdl.handle.net/10400.1/29338
dc.language.isoeng
dc.peerreviewedyes
dc.publisherSpringer Science and Business Media LLC
dc.relationCentre for Marine and Environmental Research
dc.relation.ispartofJournal of Applied Phycology
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectPhaeophyceae
dc.subjectChlorophyll
dc.subjectPigments
dc.subjectFluorescence
dc.subjectViability
dc.subjectMachine-learning
dc.titleHigh-throughput assessment of gametophyte survival and growth in the kelp Laminaria ochroleuca through autofluorescence analysiseng
dc.typejournal article
dspace.entity.typePublication
oaire.awardNumberLA/P/0101/2020
oaire.awardTitleCentre for Marine and Environmental Research
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/LA%2FP%2F0101%2F2020/PT
oaire.citation.titleJournal of Applied Phycology
oaire.fundingStream6817 - DCRRNI ID
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
person.familyNameFélix de Azeredo Pinto e Melo
person.familyNameAlves-Lima
person.familyNamede Matos Barreto
person.familyNameMonico
person.familyNameSerrao
person.familyNamePearson
person.familyNameMartins
person.givenNameFrancisca
person.givenNameCicero
person.givenNameLuís António
person.givenNameCarina
person.givenNameEster A.
person.givenNameGareth Anthony
person.givenNameNeusa
person.identifierC-6686-2012
person.identifier113536
person.identifier.ciencia-idAC1C-FB89-E69C
person.identifier.ciencia-id1D17-7538-6B6D
person.identifier.ciencia-id5B13-B26E-B1EC
person.identifier.ciencia-id3315-9919-1A52
person.identifier.ciencia-id1D1A-4459-86D8
person.identifier.orcid0000-0002-3873-4347
person.identifier.orcid0000-0003-0774-0565
person.identifier.orcid0000-0002-5619-9413
person.identifier.orcid0000-0003-1316-658X
person.identifier.orcid0000-0002-0768-464X
person.identifier.orcid0000-0003-4333-2905
person.identifier.orcid0000-0003-4333-2905
person.identifier.ridJ-3911-2013
person.identifier.scopus-author-id7004093604
person.identifier.scopus-author-id55916875600
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
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