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The cyanobacterium Agmenellum quadruplicatum PR-6 (Synechococcus sp PCC 7002) was grown turbidostatically in white light at three levels of irradiance: 20, 200, and 1260 microeinsteins per square meter per second. Phycobilisomes were isolated from each culture and analyzed by absorbance, gel electrophoresis, and electron microscopy. The ratio of phycocyanin to allophycocyanin decreased 1.8-fold from the lowest to highest irradiance. This change was due entirely to an approximately 2.5-fold decrease in one structural unit of rod domains, the complex of phycocyanin, and a 33-kilodalton linker polypeptide (LR33). For a given irradiance, phycobilisomes from cells grown on ammonium as the nitrogen source had 10 to 20% more phycocyanin than those from nitrate cultures. Total RNA was isolated from all cultures and probed with gene fragments specific to phycocyanin and allophycocyanin subunits and LR33. The relative level of RNAs encoding phycocyanin and allophycocyanin was found to vary with light intensity in parallel with the phycobiliprotein ratio. Hence, the light-harvesting capacity of phycobilisomes is directly regulated by relative levels of phycobiliprotein mRNA. The LR33 transcript occurs as a 3' extension on about 10% of phycocyanin transcripts. The ratio of RNA encoding LR33 to that encoding phycocyanin did not vary with irradiance, although the protein ratio changed 1.7- to twofold between extremes. Based on these and other observations, we propose that the LR33 protein is constitutively synthesized at a rate higher than that required to complex with available phycocyanin.

Phosphate-limited growth of the blue-green alga Agmenellum quadruplicatum resulted in the accumulation of cyanophycin granule polypeptide (CGP), which is a 1:1 co-polymer of aspartic acid and arginine. The progressive accumulation of CGP began after depletion of phosphate from the medium. CGP increased in concentration much faster than the increase in cell number. Electron microscopy indicated that both the number of cyanophycin granules per cell section and the diameter of each granule increased as phosphate starvation progressed. A marked decrease in the electron density of the inter-thylakoidal areas took place concurrently with the accumulation of CGP. At the same time a progessive decrease in the pigment concentration of cells and in the rate of nitrate uptake was observed. Thirty-two hours after phosphate depletion from the medium up to 28% of total cellular nitrogen was found in CGP.

The level of glutamine synthetase activity in Agmenellum quadruplicatum strain PR-6 was dependent on the nitrogen source used for growth and on the nutritional status of the cells. During exponential growth, glutamine synthetase activity was low in cells grown on ammonia, urea, or nitrate. During the transition from nitrogen replete to nitrogen starved growth, glutamine synthetase activity began to rise. With ammonia as a nitrogen source, glutamine synthetase activity as determined in whole cells increased from 1 nanomole per minute per milliliter during exponential growth to 22 nanomoles per minute per milliliter during severe nitrogen starvation. In cells grown on nitrate the increase was from 5 to 39 nanomoles per minute per milliliter, and in cells grown on urea the increase was from 4 to 31 nanomoles per minute per milliliter.The rise in glutamine synthetase activity corresponded with the rapid decline in the nitrogen and c-phycocyanin content of the cells. Prior to nitrogen starvation, the nitrogen content of the cells was 140, 90, and 83 micrograms nitrogen per milligram dry weight for ammonia, urea, and nitrate grown cells, respectively. During nitrogen starvation where glutamine synthetase activity was highest, the nitrogen content of cells had declined to 35 to 40 micrograms nitrogen per milligram dry weight of cells. At the same time, the c-phycocyanin content of cells dropped by 95%.

delta-Aminolevulinic acid accumulated in the culture medium when Agmenellum quadruplicatum strain PR-6 was incubated in the presence of levulinic acid, a competitive inhibitor of delta-aminolevulinic acid dehydratase, and specifically labeled glutamate and glycine. The delta-aminolevulinic acid was purified using Dowex 50W-X8 and cleaved by periodate to yield succinic acid and formaldehyde. The distribution of radioactivity in the two fragments suggested that in blue-green algae the carbon skeleton of delta-aminolevulinic acid is derived directly from glutamate. However the possibility of the pathway of delta-aminolevulinic acid synthesis, from glycine and succinyl-coenzyme A also functioning in blue-green algae was not eliminated as uptake of glycine was minimal.

A spontaneous, stable, pigmentation mutant of Nostoc sp. strain MAC was isolated. Under various growth conditions, this mutant, R-MAC, had similar phycoerythrin contents (relative to allophycocyanin) but significantly lower phycocyanin contents (relative to allophycocyanin) than the parent strain. In saturating white light, the mutant grew more slowly than the parent strain. In nonsaturating red light, MAC grew with a shorter generation time than the mutant; however, R-MAC grew more quickly in nonsaturating green light.When the parental and mutant strains were grown in green light, the phycoerythrin contents, relative to allophycocyanin, were significantly higher than the phycoerythrin contents of cells grown in red light. For both strains, the relative phycocyanin contents were only slightly higher for cells grown in red light than for cells grown in green light. These changes characterize both MAC and R-MAC as belonging to chromatic adaptation group II: phycoerythrin synthesis alone photocontrolled.A comparative analysis of the phycobilisomes, isolated from cultures of MAC and R-MAC grown in both red and green light, was performed by polyacrylamide gel electrophoresis in the presence of 8.0 molar urea or sodium dodecyl sulfate. Consistent with the assignment of MAC and R-MAC to chromatic adaptation group II, no evidence for the synthesis of red light-inducible phycocyanin subunits was found in either strain. Phycobilisomes isolated from MAC and R-MAC contained linker polypeptides with relative molecular masses of 95, 34.5, 34, 32, and 29 kilodaltons. When grown in red light, phycobilisomes of the mutant R-MAC appeared to contain a slightly higher amount of the 32-kilodalton linker polypeptide than did the phycobilisomes isolated from the parental strain under the same conditions. The 34.5-kilodalton linker polypeptide was totally absent from phycobilisomes isolated from cells of either MAC or R-MAC grown in green light.

Characteristics of nitrate reduction in terms of nitrite production in an N-methyl-N'-nitro-N-nitrosoguanidine-induced mutant of the blue-green alga Agmenellum quadruplicatum are described. Following induction of nitrate reduction a linear rate of nitrite production proportional to cell concentration was observed. Rate of nitrite production and growth rate showed similar responses to pH, temperature, and light intensity. If required, only trace amounts of carbon dioxide were necessary for nitrite production. Atmospheres of oxygen or nitrogen inhibited production of nitrite. In addition, a low but constant rate of nitrite production was observed in the dark. Nitrite production by mutant AQ-6 was studied in terms of photosynthesis. As nitrite production proceeded, rate of photosynthesis declined. Ultraviolet irradiation and 3-(3,4-dichlorophenyl)-1, 1-dimethylurea poisoning did not prevent nitrite production. The action spectrum of nitrite production was chlorophyll a-like.