An Efficient Protocol for Long-Term Preservation of Cyanobacteria


  • Mayashree B. Syiem Department of Biochemistry, North-Eastern Hill University, Shillong 793022, Meghalaya, India.
  • Amrita Bhattacharjee Department of Biochemistry, North-Eastern Hill University, Shillong 793022, Meghalaya, India.


Cyanobacteria, Nitrogenase activity, Photosynthetic activity, Respiratory activity, Long-Term Preservation


A simple modification in preparation of nutrient agar meant for agar slants resulted in a convenient and efficient matrix for long-term cyanobacterial preservation. In the modified protocol, agar concentration was increased and cyanobacterial cells were mixed rapidly in the molten agar before solidification. Solidified mixture was cut into cubes and air dried. The resulting agar flakes were stored in the dark. Periodically some flakes were inoculated in fresh medium to study regeneration of viable filaments in order to access the efficiency of the method in preserving cyanobacteria in dehydrated form. Possible outer contamination could be removed by washing the agar flakes in 1% sodium hypochlorite for one minute prior to their inoculation in fresh medium. The percentage of agar used and amount of cyanobacterial cells entrapped were the factors that influenced the period of preservation. A comparison of some biochemical and physiological characters in six regenerated cyanobacterial strains to their free-living counterparts showed that the dried agar flakes were completely reliable as preserving material for at least a period of three years. During this period the entrapped cyanobacterial cells did not need further maintenance. This process of maintaining cyanobacteria is extremely convenient as it reduces (1) input of chemicals and manpower required for maintaining cyanobacteria in batches (2) possible cross-contamination among various cyanobacteria maintained in liquid batch cultures (3) use of glassware and (4) storage space. In addition, the cells could be maintained in a near dormant state, and that minimized the chances of alterations in their native characters.


Download data is not yet available.


Acreman, J. (1994). Algae and cyanobacteria: isolation, culture and long-term maintenance. J. Ind. Microbiol., 13(3): 193–194.

Adams, D.G. (2000). Symbiotic Interactions. In: Whitton, B.A. & Potts, M. (Eds), The Ecology of Cyanobacteria. Kluwer Academic Publisher, Dordrecht, The Netherlands, pp. 523-561.

Whitton, B.A. & Carr, N.G. (1982). Cyanobacteria: current perspectives. In: Carr, N.G. & Whitton, B.A. (eds), The Biology of Cyanobacteria, Blackwell Scientific, Oxford. pp. 1–8.

Castenholz, R.W. (1981). Isolation and Cultivation of Thermophilic Cyanobacteria. In: Starr, M.P., Stolp, H., Trüper, H.G., Balows, A. & Schlegel H.G. (eds), The Prokaryotes: A Handbook on Habitats, Isolation, and Identification of Bacteria. Vol. 1, Springer-Verlag, Berlin. pp. 236-246.

Day, J.G., Benson, E.E., Harding, K., Knowles, B., Idowu, M., Bremner, D., Santos, L., Santos, F., Friedl, T., Lorenz, M., Lukesova, A., Elster, J., Lukavsky, J., Herdman, M., Rippka, R. & Hall, T. (2005). Cryopreservation and conservation of microalgae: the development of a Pan-European scientific and biotechnological resource (the COBRA project). CryoLetters, 26(4): 231–238.

De, P.K. (1939). The role of blue-green algae in nitrogen fixation in rice-fields. Proc. R. Soc. Lond. B Biol. Sci., 127: 121–139.

El-Gohary, F.A. & Nasr, F.A. (1999). Cost-effective pre-treatment of wastewater. Water Sci. Technol., 39(5): 97–103.

Hoffmann, L. (1989). Algae of terrestrial habitats. Bot. Rev., 55(2): 77–105.

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): 97–111.

Kannaiyan, S., Aruna, S.J., Kumari, S.M. & Hall, D.O. (1997). Immobilized cyanobacteria as a biofertilizer for rice crops. Intl. Conference on Applied Algology, Knysna, South Africa, April 1996. J. Appl. Phycol., 9(2): 167–174.

Kulasooriya, S.A. (1998). Cyanobacteria and Azolla as biofertilizer for rice. In: Subramanian, G.S., Kaushik, B.D. & Venkataraman, G.S. (eds), Cyanobacterial Biotechnology. Oxford and IBH Publishing Co. Pvt. Ltd, New Delhi. pp. 201–209.

Lorenz, M., Friedl, T. & Day, J.G. (2005). Perpetual maintenance of actively metabolizing microalgal cultures. In: Andersen, R.A. (Ed.), Algal Culturing Techniques. Academic Press, New York, pp. 145-156.

Mackinney, G. (1941). Absorption of Light by Chlorophyll Solutions. J. Biol. Chem., 140(2): 315–322.

Nayak, S. & Prasanna, R. (2007). Soil pH and its role in cyanobacterial abundance and diversity in rice field soils. Appl. Ecol. Environ. Res., 5(2): 103-113.

Patterson, G.M.L. (1996). Biotechnological applications of cyanobacteria. J. Sci. Ind. Res., 55: 669–684.

Prasanna, R., Dolly, D.W., Tiwari, O.N. & Singh, P.K. (2000). Microalgae-sewage interactions-implications and future prospects. Proceedings of National Symposium. “Microbes in bioremediations for an ecofriendly environment in the New Millenium”, CAS in Botany, University of Madras, Chennai.

Rai, A.N. (1990). Handbook of Symbiotic Cyanobacteria. CRC Press, Boca Raton, FL. pp. 259.

Rai, A.N., Bergman, B. & Rasmussen, U. (2002). Cyanobacteria in Symbiosis. Kluwer Academic Publisher, Dordrecht. The Netherlands. pp. 355.

Rai, A.N., Söderbäck, E. & Bergman, B. (2000). Tansley Review No. 116: Cyanobacterium-plant symbioses. New Phytol., 147(3): 449–481.

Rippka, R., Deruelles, J., Waterbury, J.B., Herdman, M. & Stanier, R.Y. (1979). Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria. J. Gen. Microbiol., 111(1): 1–61.

Robinson, S.J., Deroo, C.S. & Yocum, C.F. (1982). Photosynthetic Electron Transfer in Preparations of the Cyanobacterium Spirulina platensis. Plant Physiol., 70(1): 154–161.

Sadettin, S. & Dönmez, G. (2007). Simultaneous bioaccumulation of reactive dye and chromium(VI) by using thermophil Phormidium sp. Enzyme Microb. Technol., 41(1): 175–180.

Shah, V., Garg, N. & Madamwar, D. (2001). An integrated process of textile dye removal and hydrogen evolution using cyanobacterium, Phormidium valderianum. World J. Microbiol. Biotechnol., 17(5): 499–504.

Singh, R.N. (1961). Role of blue green algae in the nitrogen economy of Indian agriculture. Indian Council of Agricultural Research, New Delhi. pp. 175.

Skulberg, O.M. (2000). Microalgae as a source of bioactive molecules – experience from cyanophyte research. J. Appl. Phycol., 12(3): 341–348.

Smith, D. (2004). Emerging tools and technologies for isolation, conservation and preservation of microorganisms. Proceedings in NBAIM-CAB International, UK Joint Workshop on Isolation, Preservation and Conservation of Agriculturally Important Microorganisms and Use of Potential Molecular Tools for their Identification, National Bureau of Agriculturally Important Microorganisms (NBAIM), Indian Agricultural Research Institute, New Delhi, March 16-17, 2004.

Stal, L.J. (1995). Physiological ecology of cyanobacteria in microbial mats and other communities. New Phytol., 131(1): 1–32.

Stewart, W.D., Fitzgerald, G.P. & Burris, R.H. (1967). In situ studies on N2 fixation using the acetylene reduction technique. Proc. Natl. Acad. Sci. USA, 58(5): 2071–2078.

Syiem, M.B. (2005). Entrapped cyanobacteria: Implications for biotechnology. Indian J. Biotechnol., 4(2): 209–215.

Watanabe, A., Nishigaki, S. & Konishi, C. (1951). Effect of nitrogen-fixing blue-green algae on the growth of rice plants. Nature, 168: 748–749.

Whitton, B.A. (2000). Soils and rice fields. In: Whitton, B.A. & Potts, M. (Eds), The Ecology of Cyanobacteria: Their Diversity in Time and Space. Kluwer Academic Publisher, Dordrecht, pp. 233-255.

Wolk, C.P. (1988). Purification and storage of nitrogen-fixing filamentous cyanobacteria. In: Packer, L. & Glazer, A.N. (eds), Cyanobacteria, vol. 167, Methods in Enzymology, Academic Press, New York. pp. 93–95.




How to Cite

Syiem, M. B., & Bhattacharjee, A. (2010). An Efficient Protocol for Long-Term Preservation of Cyanobacteria. Journal of Advanced Laboratory Research in Biology, 1(1), 41–45. Retrieved from