Mineral requirements for the growth of Anabaena spiroides in vitro Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/fx719r28z

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  • The major nutrient requirements of the blue-green alga Anabaena spiroides Klebahn were investigated. It was hoped that the results would allow the development of a balanced culture medium in which the alga would exhibit rapid growth and retain typical morphological characteristics. The alga was isolated from Upper Klamath Lake, Oregon, and was introduced to culture in Gerloff's modification of Chu's number 10 medium. This culture solution was modified by addition of Tris (hydroxymethyl) aminomethane (Tris) and the substitution of hydrogen ferric ethylenediamine di-o-hydroxyphenylacetate (EDDHA) for ferric citrate-citric acid. Attempts to obtain bacteria-free cultures by use of antibiotics, pasteurization and ultraviolet irradiation were unsuccessful, probably because of the extremely thick gelatinous matrix surrounding the filaments. Addition of nitrogen to the culture medium was found to be unnecessary for growth. This finding strongly suggests Anabaena spiroides is capable of nitrogen-fixation but this cannot be proven until demonstrated in bacteria-free culture. Phosphorous, although required in relatively high concentrations for optimum growth, is apparently accumulated within the cells allowing the organism to survive long periods of growth in phosphorous-deficient medium. It appears that many of the major elements are either required in very minute concentrations or that their accumulation is possible. The provision of iron as the alkaline stable, chelate complex EDDHA stimulated growth and reduced the concentration of iron required in the medium. There was a definite, high requirement for calcium. The micro-nutrient requirements were not studied. The physiologically balanced medium developed provided enhanced growth of the alga so long as a nitrogen source was included. When nitrogen was eliminated growth was identical to that obtained in the original unbalanced medium. The alga requires a highly alkaline medium and is inhibited by high light intensities.
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