Extraction and detection methods for polyphosphate storage in autotrophic planktonic organisms

Eixler, Sebastian; Selig, Uwe; Karsten, Ulf

doi:10.1007/s10750-004-2406-9

Cited by 53 publications

(36 citation statements)

References 38 publications

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“…4b) and the accumulation of polyphosphate granules is maximal in stationary phase (Fig. 4e, f), while similar findings were reported by Eixler et al (2005) and Nishikawa et al (2006) under autotrophic condition.…”

Section: The Ratio Of Carbon Versus Phosphorus For the Cell Growth Ofsupporting

confidence: 87%

Phosphate assimilation by Chlorella and adjustment of phosphate concentration in basal medium for its cultivation

Shi

2008

Biotechnol Lett

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Assimilation of phosphate by Chlorella pyrenoidosa was 0.81-8.1 mg PO(4)-P/g dry weight for heterotrophic cultures and 0.81-16.1 mg/g for mixotrophic cultures. Optimal carbon:phosphorous (C/P) ratios were 206:1-2060:1 and 103:1-2060:1 for heterotrophic and mixotrophic cultivations, respectively. These requirements for phosphate for growth of C. pyrenoidosa under either heterotrophic or mixotrophic conditions are much less (6.25-62.5 or 3.12-62.5-fold at 10 g glucose/l) than its concentration in basal medium.

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Section: The Ratio Of Carbon Versus Phosphorus For the Cell Growth Ofsupporting

confidence: 87%

Phosphate assimilation by Chlorella and adjustment of phosphate concentration in basal medium for its cultivation

Shi

2008

Biotechnol Lett

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“…Boiling was performed by immersing the tubes in a floating tube rack in a beaker of boiling water; 37°C enzymatic digestions were conducted in an Eppendorf Thermomixer with gentle mixing. Some protocols were directly taken from, or inspired by, previous reports, including leaching with 0.25 M NaOH (39), simple freeze-thawing (36), boiling in Tris buffer (41), and use of EDTA (42). We first tested our methods with the Synechococcus samples and then tested those that showed most promise with Vineyard Sound samples.…”

Section: Methodsmentioning

confidence: 99%

Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

Martin

Mooy

2013

Appl Environ Microbiol

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Polyphosphate (polyP) is a ubiquitous biochemical with many cellular functions and comprises an important environmental phosphorus pool. However, methodological challenges have hampered routine quantification of polyP in environmental samples. We tested 15 protocols to extract inorganic polyphosphate from natural marine samples and cultured cyanobacteria for fluorometric quantification with 4=,6-diamidino-2-phenylindole (DAPI) without prior purification. A combination of brief boiling and digestion with proteinase K was superior to all other protocols, including other enzymatic digestions and neutral or alkaline leaches. However, three successive extractions were required to extract all polyP. Standard addition revealed matrix effects that differed between sample types, causing polyP to be over-or underestimated by up to 50% in the samples tested here. Although previous studies judged that the presence of DNA would not complicate fluorometric quantification of polyP with DAPI, we show that RNA can cause significant interference at the wavelengths used to measure polyP. Importantly, treating samples with DNase and RNase before proteinase K digestion reduced fluorescence by up to 57%. We measured particulate polyP along a North Pacific coastal-to-open ocean transect and show that particulate polyP concentrations increased toward the open ocean. While our final method is optimized for marine particulate matter, different environmental sample types may need to be assessed for matrix effects, extraction efficiency, and nucleic acid interference. Inorganic polyphosphate (polyP) is a linear polymer of three to several hundred phosphate residues that appears to be found in cells of all organisms (1). It is required for many cellular functions, including bacterial virulence (2-4), biofilm formation (4, 5), quorum sensing (4), competitive fitness in the environment (6), microbial stress responses (3,7,8), and survival during stationary phase (7, 9-11). PolyP can also act as a phosphorus (P) store in microbes. PolyP breakdown allows cells to survive during P limitation (12, 13), although some microbes transiently accumulate polyP upon nitrogen and P stress (8,14,15). Massive accumulation of polyP by certain bacteria sequesters dissolved P during biological wastewater treatment (16)(17)(18).While polyP is studied in a range of microbiological and environmental fields, the lack of easy quantitative methods has hampered progress. For example, polyP has very rarely been measured in the oceans despite increasing interest in the physiological responses of marine microbes to the ultralow P concentrations frequently found at sea and their biogeochemical implications (14,(19)(20)(21)(22)(23)(24)(25). PolyP has been found at nanomolar concentrations, comprising around 10% of total particulate P (26-28). PolyP also plays a role in redox cycling and geological sequestration of P (26, 27) and potentially in phytoplankton iron storage (29).Common analytical approaches have significant drawbacks. Phosphorus-31 nuclear magnetic resonance ...

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“…Inorganic polyphosphate (Poly-P), as an indicator of phosphorus limitation, was quantified following the procedure of Eixler et al (2005). Briefly, a 30-mL aliquot from each replicate was filtered through a pre-combusted GF/F membrane, which was then ground in 9-mL deionized water by a glass pestle.…”

Section: Nutrient-enrichment Experimentsmentioning

confidence: 99%

Nitrogen and phosphorus limitation of phytoplankton growth in different areas of Lake Taihu, China

Wang

Bin

et al. 2014

Journal of Freshwater Ecology

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Lake Taihu has attracted attention worldwide because of its extensive cyanobacterial blooms in the warm season. A better understanding of nutrient controls over phytoplankton is critical if the blooms are to be controlled. In the present study, we investigated the effects of nutrient addition on the growth of phytoplankton collected from Meiliang Bay (Station 1), Lake Center (Station 2), and Xukou Bay (Station 3) in July and September 2011. Cyanophyta was dominant in all water samples. Nevertheless, we observed higher proportions of other phyla and lower concentrations of microcystin-LR in September than in July. Further, Station 3 possessed many fewer Cyanophyta, but disproportionately higher concentrations of microcystin-LR than the other two stations. The overall phytoplankton biomass decreased in the order Station 1 > Station 2 > Station 3 and July > September. More interestingly, phosphorus-induced phytoplankton growth coincided with a distinct drop in particulate organic carbon (or nitrogen) to phosphorus ratio at Stations 2 and 3. By contrast, nitrogen, applied either alone or together with phosphorus, had negligible effect on phytoplankton growth. These results suggest that phytoplankton at Stations 2 and 3 were limited by phosphorus and no nutrient limitation occurred at Station 1. Therefore, nutrient limitation in Lake Taihu may be more dynamic and sporadic than previously thought and has implications for management of phytoplankton blooms.

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Extraction and detection methods for polyphosphate storage in autotrophic planktonic organisms

Cited by 53 publications

References 38 publications

Phosphate assimilation by Chlorella and adjustment of phosphate concentration in basal medium for its cultivation

Phosphate assimilation by Chlorella and adjustment of phosphate concentration in basal medium for its cultivation

Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

Nitrogen and phosphorus limitation of phytoplankton growth in different areas of Lake Taihu, China

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