A daily study of the diatom spring bloom at Roscoff (France) in 1985. II Phytoplankton pigment composition studied by HPLC analysis

Klein, Bert; Sournia, Alain

doi:10.3354/meps037265

Cited by 75 publications

(28 citation statements)

References 21 publications

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“…This is in agreement with historical phytoplankton pigment data at this coastal site (Klein & Sournia 1987) and with more recent data showing the dominance of chl b in the < 3 µm size fraction (M. L. Latasa unpubl.). In addition, this concurs with the large number of sequences attributed to members of the class Prasinophyceae (division Chlorophyta) retrieved from sea water samples in total extracted DNA (K. R. Romari unpubl.).…”

Section: Discussionsupporting

confidence: 92%

Application of fluorescent in situ hybridization coupled with tyramide signal amplification (FISH-TSA) to assess eukaryotic picoplankton composition

Not¹,

Simon²,

Biegala³

et al. 2002

Aquat. Microb. Ecol.

111

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Photosynthetic picoeukaryotes (phytoplankton cells with a diameter smaller than 2 to 3 µm) contribute significantly to both biomass and primary production in the oligotrophic open ocean and coastal waters, at certain times of the year. The identification of these organisms is difficult because of their small size and simple morphology, therefore hindering detailed ecological studies of their distribution and role. In this paper, we demonstrate the use of oligonucleotide probes specific to algal classes or to lower order taxa in combination with fluorescent in situ hybridization and tyramide signal amplification (FISH-TSA) to determine eukaryotic picophytoplankton diversity. Target cells were detected and enumerated using epifluorescence microscopy. The sensitivity of the technique and the specificity of the probes were tested on pure and mixed picoplanktonic strains, as well as on natural samples from the English Channel. In these samples, the community was dominated by cells belonging to the division Chlorophyta. Haptophyta, Bolidophyceae and Pelagophyceae were also detected at low abundance. The FISH-TSA method is readily applicable to the study of picoplankton diversity in natural communities. KEY WORDS: Fluorescent in situ hybridization · Tyramide signal amplification · Picoplankton · Eukaryotes · Coastal waters · DiversityResale or republication not permitted without written consent of the publisher Aquat Microb Ecol 28: 157-166, 2002 samples permit culture-independent assessments of diversity and indicate that there is a considerable number of yet uncultured species in eukaryotic picoplankton communities (Lopez-Garcia et al. 2001, Moon-van der Staay et al. 2001. The enumeration of specific taxonomic groups and the estimation of their contribution to eukaryotic picoplankton, however, are more difficult. Photosynthetic pigments have been widely used to estimate the contribution of algal classes to total chlorophyll a (chl a) biomass (Latasa & Bidigare 1998). However, this method is limited for several reasons. First, it cannot resolve diversity below the class level. Second, it is based on the assumption that the relative cellular content of diagnostic pigments is constant for a given algal group. This assumption is invalid because pigment content varies between species, as well as within species. For a given strain, it is also affected by environmental conditions (e.g. Stolte et al. 2000).Whole cell fluorescent in situ hybridization (FISH) with rRNA-targeted nucleic acid probes has been used more and more extensively to detect bacteria (Amann 1995). This method, which combines identification with quantitative determination of cell number, has been successfully applied to complex bacteria communities such as biofilms (Brümmer et al. 2000), marine sediments (Llobet-Brossa et al. 1998) and soils (Ravenschlag et al. 2000). Fluorescent rRNA probes have not been used as widely for eukaryotic phytoplankton (Simon et al. 1995, Scholin et al. 1996. Attempts to use mono-labeled oligonucleotide probes for...

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Section: Discussionsupporting

confidence: 92%

Application of fluorescent in situ hybridization coupled with tyramide signal amplification (FISH-TSA) to assess eukaryotic picoplankton composition

Not¹,

Simon²,

Biegala³

et al. 2002

Aquat. Microb. Ecol.

111

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“…The pigment diversity detected in these 2 subtidal sediments, particularly in the mud, is greater than in the phytoplankton samples collected the same year in Morlaix Bay (Klein & Sournia 1987), indicating that these pigments originate mainly from an autochthonous microphytobenthic population and not only from sedimented phytoplankton. Furthermore the chlorophyll b content relative to chlorophyll a is much higher in the sediments.…”

Section: Discussionmentioning

confidence: 64%

“…The HPLC analyses were carried out as described by Mantoura & Llewellyn (1983) with slight modifications and Klein & Sournia (1987). Reproducibility of weighing, extraction and HPLC injection was about 5 % and that of replicate injection was about 1 % .…”

Section: Methodsmentioning

confidence: 99%

Microphytobenthos from two subtidal sediments from North Brittany. II. Variations of pigment compositions and concentrations determinded by HPLC and conventional techniques

Riaux-Gobin¹,

Llewellyn²,

Klein³

1987

Mar. Ecol. Prog. Ser.

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Pigment composition and concentration of 2 temperate subtidal sediments, a mud and a sand from North Brittany, were studied using HPLC and conventional fluorometric methods. Pigment diversity, found using HPLC, was considerable, particularly in the mud (around 15 identified pigments). Pigment composition in the sand was dominated by Bacillariophyceae pigments, whereas in the mud pigments characteristic of Chlorophyceae were also found. Freeze-drying significantly reduces pigment concentrations.

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“…However, only a limited number of such studies have been conducted in estuarine and coastal waters (Klein & Sournia 1987, Roy 1989, Denant et al 1991, Bianchi et al 1993, mostly on vertically well-mixed macrotidal estuaries, and only a few have dealt with microtidal estuaries characterized by vertical salinity gradients (Denant et al 1991).…”

mentioning

confidence: 99%

Effect of salinity gradients on the distribution of phytoplankton pigments in a stratified estuary

Ahel¹,

Barlow²,

Mantoura³

1996

Mar. Ecol. Prog. Ser.

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A daily study of the diatom spring bloom at Roscoff (France) in 1985. II Phytoplankton pigment composition studied by HPLC analysis

Cited by 75 publications

References 21 publications

Application of fluorescent in situ hybridization coupled with tyramide signal amplification (FISH-TSA) to assess eukaryotic picoplankton composition

Application of fluorescent in situ hybridization coupled with tyramide signal amplification (FISH-TSA) to assess eukaryotic picoplankton composition

Microphytobenthos from two subtidal sediments from North Brittany. II. Variations of pigment compositions and concentrations determinded by HPLC and conventional techniques

Effect of salinity gradients on the distribution of phytoplankton pigments in a stratified estuary

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