Extraction methods for phycocyanin determination in freshwater filamentous cyanobacteria and their application in a shallow lake

Hajnalka Horváth, Attila W. Kovács, Caitlin Riddick, M. Présing

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Phycocyanin (PC) is one of the water-soluble accessory pigments of cyanobacteria species, and its concentration in aquatic systems is used to estimate the presence and relative abundance of blue-green algae. PC concentration and the PC/Chl-a ratio of four N2-fixing filamentous cyanobacteria strains (Cylindrospermopsis raciborskii, Anabaena spiroides, Aphanizomenon flos-aquae and Aphanizomenon issatschenkoi) common to Lake Balaton (Hungary) were determined using repeated freezing and thawing. A strong linear correlation was found between the extracted PC and Chl-a concentrations for all strains at high Chl-a concentrations (almost stable PC/Chl-a ratio in the range of 20-100 μg l-1 Chl-a). Extraction of PC and Chl-a from samples with low biomass of cyanobacteria (less than 20 μg l-1 Chl-a) proved to be unreliable using the standard protocol of freeze-thaw cycles (coefficients of variation exceeding 10-15%). In order to find an extraction method that is robust in fresh waters characterized by low algae biomass (e.g. Lake Balaton), the effectiveness of four extraction methods (repeated freeze-thaw method and homogenization with mortar and pestle, Ultrasonic, and Polytron homogenizer) were compared using C. raciborskii. It was found that the efficiency of extraction of phycocyanin was highest when a single freeze-thaw cycle was followed by sonication (25% additional yield compared with using the freeze-thaw method alone). Applying this combined method to surface water samples of Lake Balaton, a strong correlation was found between PC concentration and cyanobacterial biomass (R 2 = 0.9436), whilst the repeated freezing-thawing method found no detectable PC content. Here we show that the combined sonication/freeze-thaw method could be suitable for measuring filamentous cyanobacteria PC content, even at low concentrations; as well as for the estimation of cyanobacterial contribution to total biomass in fresh waters.

Original languageEnglish
Pages (from-to)278-286
Number of pages9
JournalEuropean Journal of Phycology
Volume48
Issue number3
DOIs
Publication statusPublished - Aug 2013

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extraction method
Cyanobacteria
cyanobacterium
lakes
lake
freeze-thaw cycle
biomass
thawing
freezing
freeze-thaw cycles
Cylindrospermopsis raciborskii
methodology
mortar
algae
water
pigment
relative abundance
Anabaena spiroides
alga
method

Keywords

  • Cylindrospermopsis raciborskii
  • extraction methods
  • fresh water
  • N-fixing filamentous cyanobacteria
  • phycocyanin
  • pigment analyses

ASJC Scopus subject areas

  • Aquatic Science
  • Plant Science

Cite this

Extraction methods for phycocyanin determination in freshwater filamentous cyanobacteria and their application in a shallow lake. / Horváth, Hajnalka; Kovács, Attila W.; Riddick, Caitlin; Présing, M.

In: European Journal of Phycology, Vol. 48, No. 3, 08.2013, p. 278-286.

Research output: Contribution to journalArticle

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abstract = "Phycocyanin (PC) is one of the water-soluble accessory pigments of cyanobacteria species, and its concentration in aquatic systems is used to estimate the presence and relative abundance of blue-green algae. PC concentration and the PC/Chl-a ratio of four N2-fixing filamentous cyanobacteria strains (Cylindrospermopsis raciborskii, Anabaena spiroides, Aphanizomenon flos-aquae and Aphanizomenon issatschenkoi) common to Lake Balaton (Hungary) were determined using repeated freezing and thawing. A strong linear correlation was found between the extracted PC and Chl-a concentrations for all strains at high Chl-a concentrations (almost stable PC/Chl-a ratio in the range of 20-100 μg l-1 Chl-a). Extraction of PC and Chl-a from samples with low biomass of cyanobacteria (less than 20 μg l-1 Chl-a) proved to be unreliable using the standard protocol of freeze-thaw cycles (coefficients of variation exceeding 10-15{\%}). In order to find an extraction method that is robust in fresh waters characterized by low algae biomass (e.g. Lake Balaton), the effectiveness of four extraction methods (repeated freeze-thaw method and homogenization with mortar and pestle, Ultrasonic, and Polytron homogenizer) were compared using C. raciborskii. It was found that the efficiency of extraction of phycocyanin was highest when a single freeze-thaw cycle was followed by sonication (25{\%} additional yield compared with using the freeze-thaw method alone). Applying this combined method to surface water samples of Lake Balaton, a strong correlation was found between PC concentration and cyanobacterial biomass (R 2 = 0.9436), whilst the repeated freezing-thawing method found no detectable PC content. Here we show that the combined sonication/freeze-thaw method could be suitable for measuring filamentous cyanobacteria PC content, even at low concentrations; as well as for the estimation of cyanobacterial contribution to total biomass in fresh waters.",
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AB - Phycocyanin (PC) is one of the water-soluble accessory pigments of cyanobacteria species, and its concentration in aquatic systems is used to estimate the presence and relative abundance of blue-green algae. PC concentration and the PC/Chl-a ratio of four N2-fixing filamentous cyanobacteria strains (Cylindrospermopsis raciborskii, Anabaena spiroides, Aphanizomenon flos-aquae and Aphanizomenon issatschenkoi) common to Lake Balaton (Hungary) were determined using repeated freezing and thawing. A strong linear correlation was found between the extracted PC and Chl-a concentrations for all strains at high Chl-a concentrations (almost stable PC/Chl-a ratio in the range of 20-100 μg l-1 Chl-a). Extraction of PC and Chl-a from samples with low biomass of cyanobacteria (less than 20 μg l-1 Chl-a) proved to be unreliable using the standard protocol of freeze-thaw cycles (coefficients of variation exceeding 10-15%). In order to find an extraction method that is robust in fresh waters characterized by low algae biomass (e.g. Lake Balaton), the effectiveness of four extraction methods (repeated freeze-thaw method and homogenization with mortar and pestle, Ultrasonic, and Polytron homogenizer) were compared using C. raciborskii. It was found that the efficiency of extraction of phycocyanin was highest when a single freeze-thaw cycle was followed by sonication (25% additional yield compared with using the freeze-thaw method alone). Applying this combined method to surface water samples of Lake Balaton, a strong correlation was found between PC concentration and cyanobacterial biomass (R 2 = 0.9436), whilst the repeated freezing-thawing method found no detectable PC content. Here we show that the combined sonication/freeze-thaw method could be suitable for measuring filamentous cyanobacteria PC content, even at low concentrations; as well as for the estimation of cyanobacterial contribution to total biomass in fresh waters.

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