Abstract
Expression of nitrogenase activity (C2H2 reduction) has been studied in the symbiotic N2-fixing cyanobacterium Anabaena azollae within the intact Azolla plant and after its isolation from the host. The cyanobacterium (blue-green alga) has been isolated successfully from the host plant by standard microbiological techniques. Optimum growth occurred in semi-solid (0.1% agar) Allen and Anion’s medium devoid of combined nitrogen. The alga has a generation time of 8–12 h when grown on atmospheric nitrogen. Addition of NO3 - or NH4 + did not change the growth rate. Actively N2-fixing filaments have a 15–18% heterocyst frequency. Under aerobic conditions, laboratory-grown A. azollae showed nitrogenase activity of 80–120 n mol C2H4 µgChla -1 h-1. The activity was linear for 2h in the light. The presence of high nitrogenase activity correlates with the observed high heterocyst frequency. Fresh intact Azolla plants showed a C2H2-reduction rate of 1.8–2 µmol C2H4g (fresh wt)-1h-1 and the activity was almost equal in light and dark, but, after 2 h dark incubation, the activity began to decline. The repression of nitrogenase activity by combined nitrogen sources viz., NO3 - or NH4 + was more pronounced in isolated A. azollae than in the whole Azolla plants.
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References
Allen M B and Arnon D I 1955 Studies on nitrogen-fixing bue-green algae. I. Growth and nitrogen fixation by Anabaena cylindrica. Plant Physiol. 30, 366–372.
Gates J E, Fisher R W, Goggin T W and Azrolan N I 1980 Antigenic differences between Anabaena azollae fresh from the Azolla fern leaf cavity and free-living cyanobacteria. Arch. Microbiol. 128, 126–129.
Gotto J W, Tabita F R and Van Baalen C 1979 Isolation and characterization of rapidly-growing marine, nitrogen-fixing strains of blue-green algae. Arch. Microbiol. 121, 155–159.
Hill D J 1975 The pattern of development of Anabaena in the Azolla-Anabaena symbiosis. Planta 122, 178–184.
Hill D J 1977 The role of Anabaena in the Azolla—Anabaena symbiosis. New Phytol. 78, 611–616.
Kobiler D, Cohen-Sharon A and Tel-or E 1981 Recognition between the N2-fixing Anabaena and the water fern Azolla. FEBS Lett. 133, 157–160.
Lis H and Sharon N 1972 Soy bean (Glycine max) agglutinin. Methods Enzymol. 28, 360–368.
Mackinney G 1941 Absorption of light by chlorophyll solutions. J. Biol. Chem. 140, 315–322.
Meeks J C, Steinberg N, Joseph C M, Enderlin C S, Jorgensen P A and Peters G A 1985 Assimilation of exogenous and dinitrogen-derived 13NH4 + by Anabaena azollae separated from Azolla caroliniana Willd. Arch. Microbiol. 142, 229–233.
Newton J W and Herman A I 1979 Isolation of cyanobacteria from the aquatic fern, Azolla. Arch. Microbiol. 120, 161–165.
Peters G A 1975 The Azolla-Anabaena azollae relationship. III. Studies on metabolic capabilities and further characterization of the symbiont. Arch. Microbiol. 103, 113–122.
Peters G A and Mayne B C 1974 The Azolla — Anabaena azollae relationship. I. Initial characterization of the association. Plant Physiol. 53, 813–819.
Peters G A, Toia R E Jr and Lough S M 1977 The Azolla-Anabaena azollae relationship. V. 15N2 fixation, acetylene reduction and H2 production. Plant Physiol. 59, 1021–1025.
Peters G A, Toia R E Jr, Raveed D and Levine N J 1978 TheAzolla-Anabaena azollae relationship. VI. Morphological aspects of the association. New Phytol. 80, 583–593.
Peters G A, Toia R E Jr, Evans W R, Crist D K, Mayne B C and Poole R E 1980 Characterization and comparisons of five N2-fixing Azolla-Anabaena associations. I. Optimization of growth conditions for biomass increase and N content in a controlled environment. Plant Cell Environ. 3, 261–269.
Ray T B, Peters G A, Toia R E Jr and Mayne B C 1978 The Azolla-Anabaena azollae relationship. VII. Distribution of ammonia-assimilating enzymes, protein and chlorophyll between host and symbiont. Plant Physiol. 62, 463–467.
Ray T B, Mayne B C, Toia R E Jr and Peters G A 1979 The Azolla-Anabaena azollae relationship. VIII. Photosynthetic characterization of the association and individual partners.Plant Physiol. 64, 791–795.
Stewart W D P 1980 Some aspects of structure and function in N2-fixing cyanobacteria. Annu. Rev. Microbiol. 34, 497–536.
Stewart W D P and Rodgers G A 1977 The cyanophyte-hepatic symbiosis. II. Nitrogen fixation and the interchange of nitrogen and carbon. New Phytol. 78, 459–471.
Stewart W D P, Fitzgerald G P and Burris R H 1967 In situ studies on N2 fixation using acetylene reduction technique.Proc. Natl. Acad. Sci., U.S.A. 58, 2071–2078.
Stewart W D P, Rowell P and Rai A N 1980 Symbiotic nitrogenfixing cyanobacteria. In Nitrogen Fixation. Eds. W D P Stewart and J R Gallon, pp 239–277. Academic Press, New York, London.
Stewart W D P, Rowell P and Rai AN 1983 Cyanobacteria-eukaryote plant symbiosis. Ann. Microbiol. (Inst. Pasteur) 134B, 205–228.
Tyagi V V S, Mayne B C and Peters G A 1980 Purification and initial characterization of phycobiliproteins from the endophytic cyanobacterium of Azolla. Arch. Microbiol. 128, 41–44.
Zimmerman W J 1987 Growth, nitrogen fixation and mass culture of isolated Anabaena azollae. Biotech. Lett. 9, 31–36.
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© 1989 Kluwer Academic Publishers
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Srivastava, M., Sharma, A., Kumar, A. (1989). Isolation and characterization of the N2-fixing symbiotic cyanobacterium Anabaena azollae . In: Skinner, F.A., Boddey, R.M., Fendrik, I. (eds) Nitrogen Fixation with Non-Legumes. Developments in Plant and Soil Sciences, vol 35. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0889-5_9
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DOI: https://doi.org/10.1007/978-94-009-0889-5_9
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