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Polyphosphate accumulation dynamics in a population of Synechocystis sp. PCC 6803 cells under phosphate overplus

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Abstract

In this study, a simple and rapid DAPI-based protocol was developed and optimized to visualize polyphosphates (polyPs) in the cyanobacterium Synechocystis sp. PCC 6803. The optimum dye concentration and incubation time were determined, and formaldehyde fixation was shown to significantly improve polyP detection in Synechocystis cells. Using the developed protocol, for the first time, it was shown that 80% of Synechocystis cells under phosphate overplus were able to accumulate phosphorus as polyP 3 min after the addition of K2HPO4. After 1 h, the number of cells with polyP began to decrease, and after 24 h, polyP granules were detected in only 30% of the cells. Thus, the Synechocystis cells appeared to be heterogeneous in their ability to accumulate and mobilize polyP. Like other photosynthetic organisms, Synechocystis synthesized less polyP in the dark than in the light. The accumulation of polyP was not inhibited under conditions of cold and heat stresses, and some cells were even able to synthesize polyP at a temperature of approximately 0 °C.

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Abbreviations

DAPI:

4′,6-diamidino-2-phenylindole

FITC:

Fluorescein isothiocyanate

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

polyP(s):

Polyphosphate(s)

GA:

Glutaraldehyde

FА:

Formaldehyde

PBS:

Phosphate-buffered saline

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Acknowledgments

In this work, the large-scale research facilities of the Collection of microalgae and cyanobacteria IPPAS (Institute of Plant Physiology RAS, Moscow, Russia) were used.

Funding

This work was supported by the Russian Scientific Foundation (grant no. 14-14-00904).

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  1. K.A. Timiryazev Institute of Plant Physiology RAS, Botanicheskaya str., 35, Moscow, 127276, Russia

    Alexander Voronkov & Maria Sinetova

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  1. Alexander Voronkov
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  2. Maria Sinetova
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Conceived and designed the experiments: MS, AV; performed the experiments: MS; performed the microscopic examination: AV; analyzed the data: AV, MS; wrote the paper: MS, AV. All authors read and approved the final manuscript.

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Correspondence to Maria Sinetova.

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Voronkov, A., Sinetova, M. Polyphosphate accumulation dynamics in a population of Synechocystis sp. PCC 6803 cells under phosphate overplus. Protoplasma 256, 1153–1164 (2019). https://doi.org/10.1007/s00709-019-01374-2

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