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Subcellular integrities in Chroococcidiopsis sp. CCMEE 029 survivors after prolonged desiccation revealed by molecular probes and genome stability assays

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Abstract

Desiccation-tolerant cells must either protect their cellular components from desiccation-induced damage and/or repair it upon rewetting. Subcellular damage to the anhydrobiotic cyanobacterium Chroococcidiopsis sp. CCMEE 029 stored in the desiccated state for 4 years was evaluated at the single-cell level using fluorescent DNA strand breakage labelling, membrane integrity and potential related molecular probes, oxidant-sensing fluorochrome and redox dye. Covalent modifications of dried genomes were assessed by testing their suitability as PCR template. Results suggest that desiccation survivors avoid/and or limit genome fragmentation and genome covalent modifications, preserve intact plasma membranes and phycobiliprotein autofluorescence, exhibit spatially-reduced ROS accumulation and dehydrogenase activity upon rewetting. Damaged cells undergo genome fragmentation, loss of plasma membrane potential and integrity, phycobiliprotein bleaching, whole-cell ROS accumulation and lack respiratory activity upon rewetting. The co-occurrence of live and dead cells within dried aggregates of Chroococcidiopsis confirms that desiccation resistance is not a simple process and that subtle modifications to the cellular milieu are required to dry without dying. It rises also intriguing questions about the triggers of dead cells in response to drying. The capability of desiccation survivors to avoid and/or reduce subcellular damage, shows that protection mechanisms are relevant in the desiccation tolerance of this cyanobacterium.

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Abbreviations

CM-H2DCFDA:

5-(and-6)-Chloromethyl-2′, dichlorodihydrofluorescein diacetate, acetyl ester

DAPI:

4′,6-Diamidino-2-phenylindole

DCF:

2′,7′-Dichlorofluorescein

DiBAC4(3):

Bis-(1,3-dibutylbarbituric acid) trimethine oxonol

FITC:

Fluorescein isothiocyanate

HIP1:

Highly iterated palindromic sequences, type1

INT:

2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-phenyl tetrazolium chloride

PCD:

Programmed cell death

STRR:

Short tandemly repeated repetitive

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling

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Acknowledgments

I thank anonymous reviewers for helpful comments. This work was funded by the Italian Space Agency (MoMa project) and the Italian Ministry of Foreign Affairs (Direzione Generale per la Promozione e Cooperazione Culturale). Thanks are due to Dr. Palma Mattioli for image analysis and Roberto Targa for skilful assistance in image editing.

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Authors and Affiliations

  1. Department of Biology, University of Rome “Tor Vergata”, Via della Ricerca Scientifica 1, 00133, Rome, Italy

    Daniela Billi

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  1. Daniela Billi
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Correspondence to Daniela Billi.

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Communicated by A. Driessen.

This paper is dedicated to the memory of E. Imre Friedmann and his wife Roseli, who pioneered researches on Chroococcidiopsis and life in desert environments.

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Billi, D. Subcellular integrities in Chroococcidiopsis sp. CCMEE 029 survivors after prolonged desiccation revealed by molecular probes and genome stability assays. Extremophiles 13, 49–57 (2009). https://doi.org/10.1007/s00792-008-0196-0

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  • DOI: https://doi.org/10.1007/s00792-008-0196-0

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