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Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii

2015-10-29Journal article Restricted access
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dc.contributor.authorChurro, Catarina
dc.contributor.authorValério, Elisabete
dc.contributor.authorVieira, Luís
dc.contributor.authorPereira, Paulo
dc.contributor.authorVasconcelos, Vitor
dc.date.accessioned2016-04-06T11:01:29Z
dc.date.available2016-11-01T01:30:09Z
dc.date.issued2015-10-29
dc.description.abstractMolecular studies on cyanobacteria often involve filtering and freezing of samples leading to loss of cell morphological features. Methanol is often used in preservation of biological materials in association with other fixatives. This study evaluates the application of methanol in the preservation of DNA for molecular studies as well as for the preservation of cell morphology for morphometric analysis in filamentous cyanobacteria. In the present study, both culture and environmental bloom samples were preserved using a cold methanol dehydration series (50, 70, and 100 %) and stored at −20 °C for up to 2 years. The DNA quantity and quality, nucleotide sequence retrieval, and real-time PCR quantification were analyzed over time. Morphometric cell analysis was performed on preserved samples. Results show that the DNA extracted from samples preserved up to 6 months was successfully quantified by real-time PCR. After that period, the DNA quantity decreased with the preservation time. Nevertheless, we were able to detect/amplify the target fragment in samples preserved up to 2 years. The DNA sequence and cell morphology were also maintained during the preservation time. Thus, methanol preservation is an adequate method to preserve molecular information and morphological features after long storage periods.pt_PT
dc.description.sponsorshipThis research was partially supported by the European Regional Development Fund (ERDF) through the COMPETE - Operational Competitiveness Program - and FCT - Foundation for Science and Technology national funds through projects PPCDT/AMB/67075/2006 and PEst-C/MAR/LA0015/2013, the Post-Doc research grant SFRH/BPD/75922/2011 to E. Valério and Ph.D. research grant SFRH/BD65706/2009 to C. Churro.pt_PT
dc.identifier.citationJ. Appl. Phycol. 2015. Epub 2015 Oct 29. doi: 10.1007/s10811-015-0732-0).pt_PT
dc.identifier.doi10.1007/s10811-015-0732-0pt_PT
dc.identifier.issn0921-8971
dc.identifier.urihttp://hdl.handle.net/10400.18/3774
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherSpringer Verlagpt_PT
dc.relationThe application of quantitative real-time PCR to studies of the abundance and toxicity of cyanobacteria in Portuguese potable and recreational water supplies
dc.relation.publisherversionhttp://link.springer.com/article/10.1007%2Fs10811-015-0732-0
dc.subjectCyanobacteriapt_PT
dc.subjectLugol’s iodinept_PT
dc.subjectMethanolpt_PT
dc.subjectPreservationpt_PT
dc.subjectReal-time qPCRpt_PT
dc.subjectrpoC1pt_PT
dc.subjectÁgua e Solopt_PT
dc.titleEvaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhiipt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleThe application of quantitative real-time PCR to studies of the abundance and toxicity of cyanobacteria in Portuguese potable and recreational water supplies
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876-PPCDTI/PTDC%2FAMB%2F67075%2F2006/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/COMPETE/PEst-C%2FMAR%2FLA0015%2F2013/PT
oaire.citation.endPage11pt_PT
oaire.citation.startPage1pt_PT
oaire.citation.titleJournal of Applied Phycologypt_PT
oaire.fundingStream5876-PPCDTI
oaire.fundingStreamCOMPETE
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsembargoedAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublication08dd2d8c-1680-4e89-9750-7c8987aa1a2e
relation.isProjectOfPublication93b38b87-6e5d-4154-b69f-355e4ee7efaf
relation.isProjectOfPublication.latestForDiscovery08dd2d8c-1680-4e89-9750-7c8987aa1a2e

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