Skip to main content Accessibility help

Restart capability of resting-states of Euglena gracilis after 9 months of dormancy: preparation for autonomous space flight experiments

  • Sebastian M. Strauch (a1), Ina Becker (a1), Laura Pölloth (a1), Peter R. Richter (a1), Ferdinand W. M. Haag (a1), Jens Hauslage (a2) and Michael Lebert (a1)...


Dormant states of organisms are easier to store than the living state because they tolerate larger variations in temperature, light, storage space etc., making them attractive for laboratory culture stocks and also for experiments under special circumstances, especially space flight experiments. Like several other organisms, Euglena gracilis is capable of forming desiccation tolerant resting states in order to survive periods of unfavourable environmental conditions. In earlier experiments it was found that dormant Euglena cells must not become completely desiccated. Some residual moisture is required to ensure recovery of the resting states. To analyse the water demand in recovery of Euglena resting states, cells were transferred to a defined amount of cotton wool (0.5 g). Subsequently different volumes of medium (1, 2, 3, 4, 5, 8, 10 and 20 ml) were added in order to supply humidity; a control was set up without additional liquid. Samples were sealed in transparent 50 ml falcon tubes and stored for 9 months under three different conditions:

  • Constant low light conditions in a culture chamber at 20°C,
  • In a black box, illuminated with short light emitting diode-light pulses provided by joule thieves and
  • In darkness in a black box.

After 9 months, cells were transferred to fresh medium and cell number, photosynthetic efficiency and movement behavior was monitored over 3 weeks. It was found that cells recovered under all conditions except in the control, where no medium was supplied. Transcription levels of 21 genes were analysed with a Multiplex-polymerase chain reaction. One hour after rehydration five of these genes were found to be up-regulated: ubiquitin, heat shock proteins HSP70, HSP90, the calcium-sensor protein frequenin and a distinct protein kinase, which is involved in gravitaxis. The results indicate a transient general stress response of the cells.


Corresponding author


Hide All
Allen, A.E., LaRoche, J., Maheswari, U., Lommer, M., Schauer, N., Lopez, P.J., Finazzi, G., Fernie, A.R. & Bowler, C. (2008). Whole-cell response of the pennate diatom Phaeodactylum tricornutum to iron starvation. PNAS 105(30), 1043810443.
Allen, M.J. & Crane, A.E. (1976). Null potential voltammetry – an approach to the study of plant photosystems. Bioelectrochem. Bioenergetics 3, 8491.
Alpert, P. (2006). Constraints of tolerance: why are desiccation-tolerant organisms so small or rare? J. Exp. Biol. 209(Pt. 9), 15751584.
Checcucci, A. (1976). Molecular sensory physiology of Euglena. Naturwissenschaften 63(9), 412417.
Ciechanover, A. (1994). The ubiquitin-proteasome proteolytic pathway. Cell 79(1), 1321.
Farrant, J.M., Cooper, K., Hilgart, A., Abdalla, K.O., Bentley, J., Thomson, J.A., Dace, H.J.W., Peton, N., Mundree, S.G. & Rafudeen, M.S. (2015). A molecular physiological review of vegetative desiccation tolerance in the resurrection plant Xerophyta viscosa (Baker). Planta 242(2), 407426.
Finley, D., Özkaynak, E. & Varshavsky, A. (1987). The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses. Cell 48(6), 10351046.
Häder, D.-P., Richter, P. & Lebert, M. (2006). Signal transduction in gravisensing of flagellates. Signal Transduction 6(6), 422431.
Hill, D.R., Peat, A. & Potts, M. (1994) Biochemistry and structure of the glycan secreted by desiccation-tolerant Nostoc commune (cyanobacteria). Protoplasma 182, 126148, from On CD1.
Hu, C., Zhang, D., Huang, Z. & Liu, Y. (2003) The vertical microdistribution of cyanobacteria and green algae within desert crusts and the development of the algal crusts. Plant Soil 257(1), 97111.
Ingram, J. & Bartels, D. (1996) The molecular basis of dehydration tolerance in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47, 377403.
Jayakumar, A., Hwang, S.J., Fabiny, J.M., Chinault, A.C. & Barnes, E. M. Jr. (1989) Isolation of an ammonium or methylammonium ion transport mutant of Escherichi coli and complementation of the cloned gene. J. Bacteriol. 171, 9961001, from On CD1.
Kalaji, H.M. et al. (2014). Frequently asked questions about in vivo chlorophyll fluorescence: practical issues. Photosynth. Res 122(2), 121158.
Kaprelyants, A.S., Gottschal, J.C. & Kell, D.B. (1993). Dormancy in non-sporulating bacteria. FEMS Microbiol. Lett. 104(3–4), 271286.
Knight, H. (2005). Calcium signaling during abiotic stress in plants. Int. Rev. Cytol. 195, 269324.
Kosová, K., Vítámvás, P., Prášil, I.T. & Renaut, J. (2011). Plant proteome changes under abiotic stress – Contribution of proteomics studies to understanding plant stress response. J. Proteomics 74(8), 13011322.
Lebert, M. & Häder, D.-P. (1999). Image analysis: a versatile tool for numerous applications. G.I.T. Imaging Microscopy 1, 56.
LeBlanc, J.C., Goncalves, E.R. & Mohn, W.W. (2008). Global response to desiccation stress in the soil actinomycete Rhodococcus jostii RHA1. Appl. Environ. Microbiol. 74(9), 26272636, viewed 28 November 2016.
Li, H., Rao, B., Wang, G., Shen, S., Li, D., Hu, C. & Liu, Y. (2014). Spatial heterogeneity of cyanobacteria-inoculated sand dunes significantly influences artificial biological soil crusts in the Hopq Desert (China). Environ. Earth Sci. 71(1), 245253.
Malik, K.A. (1990). A simplified liquid-drying method for the preservation of microorganisms sensitive to freezing and freeze-drying. J. Microbiol. Methods 12(2), 125132.
Malik, K.A. (1993). Preservation of unicellular gree algae by a liquid-drying method. J. Microbiol. Methods 18(1), 4149, from
Malik, K.A. (1995) A convenient method to maintain unicellular green algae for long times as standing liquid cultures. J. Microbiol. Methods 22, 221227.
Nasir, A. et al. (2014). The influence of microgravity on Euglena gracilis as studied on Shenzhou 8. Plant Biol. (Stuttgart, Germany) 16 (Suppl. 1), 113119.
O'Mahony, P.J. & Oliver, M.J. (1999). The involvement of ubiquitin in vegetative desiccation tolerance. Plant Mol. Biol. 41, 657667, viewed 28 November 2016.
Poovaiah, B.W. & Reddy, A.S.N. (1990). Turnover of inositol phospholipids and calcium-dependent protein phosphorylation in signal transduction. Inositol Metab. Plants 9, 335349.
Rao, B., Liu, Y., Wang, W., Hu, C., Dunhai, L. & Lan, S. (2009). Influence of dew on biomass and photosystem II activity of cyanobacterial crusts in the Hopq Desert, northwest China. Soil Biol. Biochem. 41(12), 23872393.
Reddy, A.S., Ali, G.S., Celesnik, H. & Day, I.S. (2012). Coping with stresses: roles of calcium- and calcium/calmodulin-regulated gene expression. Plant Cell 23(6), 20102032, viewed 29 November 2016.
Richter, P.R., Schuster, M., Meyer, I., Lebert, M. & Häder, D.-P. (2006). Indications for acceleration-dependent changes of membrane potential in the flagellate Euglena gracilis . Protoplasma 229(2–4), 101108.
Richter, P.R., Schuster, M., Lebert, M., Streb, C. & Häder, D.-P. (2007). Gravitaxis of Euglena gracilis depends only partially on passive buoyancy. Adv. Space Res. 39(7), 12181224.
Rio, D.C., Ares, M. Jr., Hannon, G.J. & Nilsen, T.W. (2010). RNA: A laboratory manual. Cold Spring Harbor Laboratory Press, New York.
Rojas-Triana, M., Bustos, R., Espinosa-Ruiz, A., Prat, S., Paz-Ares, J. & Rubio, V. (2013). Roles of Ubiquitination in the control of phosphate starvation responses in plants F. J. Integrative Plant Biol. 55(1), 4053.
Rosowski, J.R. (1977). Development of mucilaginous surfaces in Euglenoids. II. Flagellated, creeping and palmelloid cells of Euglena. J. Phycol. 13(4), 323328.
Sandgren, C.D. (1988) Growth and reproductive strategies of freshwater phytoplankton. 1st edn. Cambridge University Press, Cambridge.
Scherer, S. & Potts, M. (1989). Novel water stress protein from a desiccation-tolerant cyanobacterium. Purification and partial characterization. J. Biol. Chem. 264, 1254612553.
Starr, R.C. (1964) The culture collection of Algae at Indiana University. Am. J. Bot. 51(9), 10131044, from
Tahedl, H. & Häder, D.P. (2001). Automated biomonitoring using real time movement analysis of Euglena gracilis . Ecotoxicol. Environ. Saf. 48(2), 161169.
Tahedl, H.A. (2000) Entwicklung eines vollautomatischen Analysesystems für ökotoxikologische Untersuchungen. Doctoral Thesis, Ökophysiologie der Pflanzen, Friedrich-Alexander University, Erlangen-Nürnberg.
Takenaga, S., Kondo, T., Nazeri, S., Tamura, Y., Tokunaga, M., Tsuyama, S., Miyatake, K. & Nakano, Y. (1997). Accumulation of trehalose as a compatible solute under osmotic stress in Euglena gracilis Z. J. Eukaryot. Microbiol. 44(6), 609613.
Timperio, A.M., Egidi, M.G. & Zolla, L. (2008). Proteomics applied on plant abiotic stresses: role of heat shock proteins (HSP). J. Proteomics 71(4), 391411.
Toroser, D. & Huber, S.C. (1997). Protein phosphorylation as a mechanism for osmotic-stress activation of sucrose-phosphate synthase in spinach leaves. Plant Physiol. 114, 947955, viewed 28 November 2016.
Vaishampayan, A., Sinha, R.P., Häder, D.-P., Dey, T., Gupta, A.K., Bhan, U. & Rao, A.L. (2001). Cyanobacterial biofertilizers in rice agriculture. Bot. Rev. 67, 453516.
Vinocur, B. & Altman, A. (2005). Recent advances in engineering plant tolerance to abiotic stress: achievements and limitations. Curr. Opin. Biotechnol. 16(2), 123132.
Wang, W., Liu, Y., Li, D., Hu, C. & Rao, B. (2009). Feasibility of cyanobacterial inoculation for biological soil crusts formation in desert area. Soil Biol. Biochem. 41(5), 926929.
Wang, W., Vinocur, B., Shoseyov, O. & Altman, A. (2004). Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci. 9(5), 244252.
Xie, Z., Liu, Y., Hu, C., Chen, L. & Li, D. (2007). Relationships between the biomass of algal crusts in fields and their compressive strength. Soil Biol. Biochem. 39(2), 567572.


Restart capability of resting-states of Euglena gracilis after 9 months of dormancy: preparation for autonomous space flight experiments

  • Sebastian M. Strauch (a1), Ina Becker (a1), Laura Pölloth (a1), Peter R. Richter (a1), Ferdinand W. M. Haag (a1), Jens Hauslage (a2) and Michael Lebert (a1)...


Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed