Enzymatic Permeabilization of the Thecate Dinoflagellate<i>Alexandrium minutum</i>(Dinophyceae) Yields Detection of Intracellularly Associated Bacteria via Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization

Palacios, Lucía; Marı́n, Irma

doi:10.1128/aem.01144-07

Cited by 11 publications

(8 citation statements)

References 29 publications

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“…), and we hypothesized that, if present, this may have impeded access to cell contents by the FISH probes. However, in this case, the enzymatic treatments tested should have increased the proportion of hybridized cells, as happens, for example in thecate dinoflagellates (Palacios and Marín ), but it did not. Alternately, this nonhybridizing fraction may be a resting or cyst stage with low ribosomal activity and little 18S rRNA, such that no signal was observed even with CARD‐FISH.…”

Section: Discussionmentioning

confidence: 86%

Distribution and Diversity of a Protist PredatorCryothecomonas(Cercozoa) in Arctic Marine Waters

Thaler

Lovejoy

2012

J Eukaryotic Microbiology

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Heterotrophic nanoflagellates (HNFs) are key components in microbial food webs, potentially influencing community composition via top‐down control of their favored prey or host. Marine cercozoan Cryothecomonas species are parasitoid and predatory HNFs that have been reported from ice, sediments, and the water column. Although Cryothecomonas is frequently reported from Arctic and subarctic seas, factors determining its occurrence are not known. We investigated the temporal and geographic distribution of Cryothecomonas in Canadian Arctic seas during the summer and autumn periods from 2006 to 2010. We developed a Cryothecomonas‐specific fluorescent in situ hybridization (FISH) probe targeting ribosomal 18S rRNA to estimate cell concentrations in natural and manipulated samples. Comparison of simple and partial correlation coefficients showed that salinity, depth, and overall community biomass are important factors determining Cryothecomonas abundance. We found no evidence of parasitism in our samples. Hybridized cells included individuals smaller than any formally described Cryothecomonas, suggesting the presence of novel taxa or unknown life stages in this genus. A positive relationship between Cryothecomonas abundance and ice and meltwater suggests that it is a sensitive indicator of ice melt in Arctic water columns.

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Section: Discussionmentioning

confidence: 86%

Distribution and Diversity of a Protist PredatorCryothecomonas(Cercozoa) in Arctic Marine Waters

Thaler

Lovejoy

2012

J Eukaryotic Microbiology

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“…All cultures were monitored for 15 d, and sampled every 2 days for cell counting by light microscopy. One milliliter of L. polyedrum cells from V-R and V-L cultures were harvested at lag (d2), exponential (d8) and stationary (d15) phase and fixed with paraformaldehyde-PBS at a final concentration of 1% for 12 h at 4 • C. For attached bacteria, fixed cells were immobilized onto an 8.0 µm pore size, 25 mm-diameter Nucleopore filters (Whatman International, Ltd., Maidstone, England), and rinsed with phosphate-buffered saline (PBS, 0.1 M NaCl, 2 mM KCl, 4 mM Na 2 HPO 4 , pH 8.1; Palacios and Marín, 2008). For freeliving bacteria, the fraction which passed through an 8.0 µm pore size filter was collected on 0.2 µm pore size, 25 mm-diameter Nucleopore filters (Whatman International, Ltd., Maidstone, England) and rinsed with PBS.…”

Section: Sampling and Cell Fixationmentioning

confidence: 99%

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

et al. 2018

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Recent research has shown that in aquatic systems pairs of prokaryote and eukaryote species exercise symbiotic exchanges of metabolites that are essential for the proliferation of either species. Using dinoflagellate Lingulodinium polyedrum cultures and a factorial design, we examined its growth at different concentrations of vitamin B 1 (thiamine) and B 12 (cobalamin). When both vitamins were at their lowest concentrations tested, 0.033 pM of B 1 and 0.053 pM of B 12 the growth was limited. When axenic L. polyedrum was co-cultured with the bacterium Dinoroseobacter shibae, a known B 1 and B 12 producer, then L. polyedrum grew at the same rate as in culture media supplemented with B 1 and B 12. In the L. polyedrum vitamin-limited culture (V-L), the abundance of attached and free-living D. shibae was higher than in the vitamin-replete (V-R) culture. In the V-R and V-L co-cultures the measured particulate B 12 (pB 12) concentration of attached and free-living D. shibae were in the range of 4.7 × 10 −19 to 3 × 10 −18 and 8.4 × 10 −21 to1.2 × 10 −19 (mol cell −1), respectively. Without B 12 or B 7 (biotin) added to the culture medium of a co-culture of L. polyedrum and D. shibae, the measured dissolved B 12 (dB 12) concentration was more than 60 pM higher than necessary for unlimited growth rates of L. polyedrum. In the same culture we measured B 7 in the L. polyedrum particulate fraction (pB 7 ; 4.7 × 10 −19 to 9.4 × 10 −19 mol cell −1). We suggest that in response to the production of B 1 and B 12 by D. shibae to supply L. polyedrum requeriments, the latter produced B 7 , which is required by D. shibae, and in our culture was only produced by L. polyedrum when D. shibae was present. We propose that D. shibae can control L. polyedrum through the release of B 1 and B 12 , and L. polyedrum can control D. shibae through the release of B 7. D. shibae is also auxotroph for niacin and 4-amino-benzoic acid, not provided by the culture medium. Therefore, L. polyedrum might affect a similar control through the release of these specific compounds and organic substrate necessary for the growth of D. shibae.

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“…The microalgae which were observed in Fig. a neither showed autofluoresce, thus are not photosynthetic, nor did they hybridize with the eukaryotic probe, as the permeabilization steps that were followed were not stringent enough to permeabilize microalgal cells (Palacios and Marín ). A screening of biofilm samples from both towers with FISH probes (Table ) revealed the presence of Alpha, Delta and Gammaproteobacteria at class level, with inconclusive detection of Betaproteobacteria due to a nonspecific signal.…”

Section: Resultsmentioning

confidence: 99%

The microbial community of a biofilm contact reactor for the treatment of winery wastewater

2018

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Enzymatic Permeabilization of the Thecate DinoflagellateAlexandrium minutum(Dinophyceae) Yields Detection of Intracellularly Associated Bacteria via Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization

Cited by 11 publications

References 29 publications

Distribution and Diversity of a Protist PredatorCryothecomonas(Cercozoa) in Arctic Marine Waters

Distribution and Diversity of a Protist PredatorCryothecomonas(Cercozoa) in Arctic Marine Waters

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

The microbial community of a biofilm contact reactor for the treatment of winery wastewater

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