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Combining flow cytometry and real-time PCR methodology to demonstrate consumption in Prymnesium parvum
Linnéuniversitetet, Fakultetsnämnden för naturvetenskap och teknik, Institutionen för naturvetenskap, NV.
Linnéuniversitetet, Fakultetsnämnden för naturvetenskap och teknik, Institutionen för naturvetenskap, NV.
Linnéuniversitetet, Fakultetsnämnden för naturvetenskap och teknik, Institutionen för naturvetenskap, NV.
2010 (Engelska)Ingår i: Journal of the American Water Resources Association, ISSN 1093-474X, E-ISSN 1752-1688, Vol. 46, s. 133-143Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Harmful algal bloom species can persist in the environment, impacting aquatic life and human health. One of the mechanisms by which some harmful algal bloom species are able to persist is by consumption of organic particles. Methods to demonstrate and measure consumption can yield insight into how populations thrive. Here, we combine flow cytometry and real-time PCR to demonstrate consumption of a cryptophyte species (Rhodomonas sp.) by a toxic mixotrophic haptophyte (Prymnesium parvum). Using flow cytometry, the feeding frequency of a population of P. parvum cells was calculated using the phycoerythrin (PE) fluorescence signal from Rhodomonas sp. and the fluorescence of an acidotropic probe labeling the food vacuoles. Feeding frequency increased in the beginning of the experiment and then began to decline, reaching a maximum of 47.5% of the whole P. parvum population after 212 min. The maximum number of consumed Rhodomonas sp. cells was 0.8 per P. parvum cell, and occurred after 114 min corresponding to an ingestion rate of 0.4 Rhodomonas sp. cells/P. parvum/h. Cells from the feeding P. parvum population were sorted, washed, and subjected to a real-time PCR assay targeting the cryptophyte 18S locus. There was a correlation between cycle threshold (Ct) values and number of consumed prey cells calculated by fluorescence. Overall, this study shows that flow cytometric analysis, of the acidotropic probe and prey pigments, is an efficient and rapid tool in enumerating food vacuoles and the number of prey cells consumed. Furthermore, we suggest that real-time PCR can be applied to cells sorted by flow cytometry, thus allowing for the detection and potential quantification of the targeted prey cells.

Ort, förlag, år, upplaga, sidor
2010. Vol. 46, s. 133-143
Nationell ämneskategori
Ekologi
Forskningsämne
Ekologi, Akvatisk ekologi
Identifikatorer
URN: urn:nbn:se:lnu:diva-2159DOI: 10.1111/j.1752-1688.2009.00397.xISI: 000274175400012Scopus ID: 2-s2.0-77952926818OAI: oai:DiVA.org:lnu-2159DiVA, id: diva2:309209
Tillgänglig från: 2010-04-06 Skapad: 2010-04-06 Senast uppdaterad: 2023-08-25Bibliografiskt granskad

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Brutemark, AndreasF de Carvalho, WandersonGranéli, Edna

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Brutemark, AndreasF de Carvalho, WandersonGranéli, Edna
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Journal of the American Water Resources Association
Ekologi

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