Journal of Applied Phycology

, 21:737 | Cite as

Cyanobacterial akinete induction and its application as biofertilizer for rice cultivation

  • Sasidhorn Innok
  • Somporn Chunleuchanon
  • Nantakorn Boonkerd
  • Neung Teaumroong


Nostoc sp. VICCR1-1 was induced in order to form akinetes on the basis of nutrient modification. Phosphorus and iron were found to be the critical for akinete differentiation, especially when both elements were omitted. The number of akinete cells increased up to 20% when compared with culturing in BG110 medium (without N source). In addition, CaCl2 played a role in heterocyst differentiation, and was able to induce heterocyst ranging between 30% and 46%. In order to prepare akinetes as inoculum, the dried form of akinetes was prepared by mixing it with montmorillonite clay. The inoculum with the amount of 2.8 × 106 cells m−2 was applied to rice (Oryza sativa) fields. After harvesting, the grain yields from chemical N fertilizer, vegetative cells, and akinete inoculum treatments were not significantly different. To monitor the persistence of Nostoc sp. VICCR1-1 after harvesting, the most probable number-denaturing gradient gel electrophoresis technique using 16S rRNA gene was employed. The results indicated that the remaining population is at 2.5 × 105 and 1.62 × 106 cells m−2 in treatments supplied with vegetative cells and akinete inocula, respectively. Akinete induction might be one of the appropriate approaches for producing cyanobacterial inoculum.


Nostoc sp. Akinete induction Akinete inoculum Montmorillonite clay Rice cultivation MPN-DGGE 



This work was fully supported by Suranaree University of Technology. The authors thank Dr. Issra Pramoolsook for advice and comments on the manuscript.


  1. Canini A, Civitareale P, Marini S, Caiola MG, Rotilio G (1992) Purification of iron superoxide dismutase from the cyanobacterium Anabaena cylindrica Lemn. and localization of the enzyme in heterocysts by immunogold labeling. Planta 187:438–444, doi: 10.1007/BF00199961 CrossRefGoogle Scholar
  2. Chairin E (2002) Evaluation of uptake of nitrogen from cyanobacteria in rice plant using 15N. MSc Thesis, Chiangmai University, Thailand, 113 ppGoogle Scholar
  3. Chairin E, Prasatsrisupab J, Choonluchanon S (2004) Evaluation of nitrogen uptake by rice from 15N labeled cyanobacteria. Biotechnol Sustain Util Resour Trop 17:195–201Google Scholar
  4. Hall DO, Kannaiyan S, van der Leij M (2002) Ammonia production in rice paddies using immobilized cyanobacteria. In: Kannaiyan S (ed) Biotechnology of biofertilizer. Narosa, New Delhi, India, pp 370–375Google Scholar
  5. Innok S, Matsumura M, Boonkerd N, Teaumroong N (2005) Detection of Microcystis in lake sediment using molecular genetic techniques. World J Microbiol Biotechnol 21:1559–1568, doi: 10.1007/s11274-005-7893-y CrossRefGoogle Scholar
  6. Janse I, Meima M, Kardinaal EA, Zwart G (2003) High-resolution differentiation of cyanobacteria by using rRNA-internal transcribed spacer denaturing gradient gel electrophoresis. Appl Environ Microbiol 69:6634–6643, doi: 10.1128/AEM.69.11.6634-6643.2003 CrossRefPubMedGoogle Scholar
  7. Kim EJ, Chung HJ, Suh B, Hah YC, Roe JH (1998) Expression and regulation of the sodF gene encoding iron- and zinc-containing superoxide dismutase in Streptomyces coelicolor. J Bacteriol 180:2014–2020PubMedGoogle Scholar
  8. Mishra U, Pabbi S (2004) Cyanobacteria: a potential biofertilizer for rice. Resonance 9:6–10CrossRefGoogle Scholar
  9. Muyzer G (1999) DGGE/TGGE a method for identifying genes from natural ecosystems. Curr Opin Microbiol 2:317–322, doi: 10.1016/S1369-5274(99)80055-1 CrossRefPubMedGoogle Scholar
  10. Nohr RS (1990) Immobilized blue-green algae. US Patent 4921803, 5 January 1990Google Scholar
  11. Olli K, Kangro K, Kabel M (2005) Akinete production of Anabaena lemmermannii and A. cylindrica (cyanophyceae) in natural populations of N- and P- limited coastal mesocosms. J Phycol 41:1094–1098, doi: 10.1111/j.1529-8817.2005.00153.x CrossRefGoogle Scholar
  12. Richmond A (1986) CRC handbook of microalgal mass culture. CRC, Boca RatonGoogle Scholar
  13. Roger PA, Reynaud PA (1982) Free-living blue-green algae in tropical soils. In: Dommergues YR, Diem HG (eds) Microbiology of tropical soils and plant productivity. The Hague, Martinus Nijhoff, pp 147–168Google Scholar
  14. Roger PA, Santiago-Ardales S, Reddy PM, Watanabe I (1987) The abundance of heterocystous blue-green algae in rice fields. Biol Fertil Soils 4:98–105Google Scholar
  15. Sutherland JM, Herdman M, Stewart WDP (1979) Akinetes of the cyanobacterium Nostoc PCC7524: macromolecular composition, structure and control of differentiation. J Gen Microbiol 115:273–287Google Scholar
  16. Teaumroong N, Innok S, Chunleuchanon S, Boonkerd N (2002) Diversity of nitrogen-fixing cyanobacteria under various ecosystems of Thailand: I. Morphology, physiology and genetic diversity. World J Microbiol Biotechnol 18:673–682, doi: 10.1023/A:1016812116538 CrossRefGoogle Scholar
  17. Teske A, Wawer C, Muyzer G, Ramsing NB (1996) Distribution of sulfate-reducing bacteria in a stratified Fjord (Mariager Fjord, Denmark) as evaluated by most-probable number counts and denaturing gradient gel electrophoresis of PCR-amplified ribosomal DNA fragments. Appl Environ Microbiol 62:1405–1415PubMedGoogle Scholar
  18. Torrecilla I, Leganes F, Bonilla I, Fernandez-Pinas F (2004) A calcium signal is involved in heterocyst differentiation in the cyanobacterium Anabaena sp. PCC7120. Microbiol 150:3731–3739, doi: 10.1099/mic.0.27403-0 CrossRefGoogle Scholar
  19. Watanabe A, Yamamoto Y (1971) Algal nitrogen fixation in the tropics. Plant Soil (special volume):403–413, doi: 10.1007/BF02661867 CrossRefGoogle Scholar
  20. Wolk CP, Ernst A, Elhai J (1994) Heterocyst metabolism and development. In: Bryant DA (ed) The molecular biology of cyanobacteria, vol. 1. Kluwer Academic, The Netherlands, pp 769–823Google Scholar
  21. Wong FC, Meeks JC (2002) Establishment of a functional symbiosis between the cyanobacterium Nostoc punctiforme and the bryophyte Anthoceros punctatus requires genes involved in nitrogen control and initiation of heterocyst differentiation. Microbiol 148:315–323Google Scholar
  22. Yang CH, Crowley DE (2002) Rhizosphere microbial community structure in relation to root location and plant iron nutritional status. Appl Environ Microbiol 66:345–351CrossRefGoogle Scholar
  23. Zhao Y, Shi Y, Zhao W, Huang X, Wang D, Brown N, Brand J, Zhao J (2005) CcbP, a calcium-binding protein from Anabaena sp. PCC7120, provides evidence that calcium ions regulate heterocyst differentiation. Proc Natl Acad Sci USA 102:5744–5748, doi: 10.1073/pnas.0501782102 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Sasidhorn Innok
    • 1
  • Somporn Chunleuchanon
    • 2
  • Nantakorn Boonkerd
    • 1
  • Neung Teaumroong
    • 1
  1. 1.School of Biotechnology, Institute of Agricultural TechnologySuranaree University of TechnologyNakhon RatchasimaThailand
  2. 2.Department of Soil Sciences and Conservation, Faculty of AgricultureChiang Mai UniversityChiang MaiThailand

Personalised recommendations