INTERACTION OF NITROGEN FIXATION AND PHOSPHORUS LIMITATION IN APHANIZOMENON FLOS-AQUAE (CYANOPHYCEAE)1

The effect of simultaneous nitrogen fixation and phosphorus limitation on the physiological adaptation and growth performance of Aphanizomenon flos-aquae (L.) Ralfs PCC 7905 was studied in continuous culture. In the absence of ammonia, N₂ fixation occurred and the maximum growth rate (as determined in diluted batch cultures) was lower. However, no distinction could be made between the steady-state N uptake rates (based on cellular N contents) of N₂-fixing cells and cells grown with ammonia. At the higher dilution rates, the residual P concentration increased with increasing dilution rate, more so under N₂-fixing conditions, compared to the cultures grown in the presence of ammonia. More generally, the yield of biomass per consumed P, as the biomass concentration itself, decreased with increasing dilution rate, and both were lower under N₂-fixing conditions. The restricted biomass production under N₂-fixing conditions suggests that reduction of N loading may benefit lake restoration projects. The influence of N₂-fixation on the severity of P limitation is discussed in terms of metabolic control analysis. From the increase of the residual P concentration on switching from ammonium to N₂-fixing conditions, it is deduced that under N₂-fixing and P-limited conditions, control of growth is shared by N and P metabolism.     

Malat-dehydrogenase isoenzymbanden als potentielles chemotaxonomisches kriterium für cyanophyceen-species

Malate dehydrogenase isoenzymes from eight cyanophycean species were investigated with polyacrylamide disc gel electrophoresis. Using 7% acrylamide the pherograms from each species showed 5–8 zones with malate dehydrogenase activity. It was demonstrated that in Anabaena flos-aquae there are 8 isoenzyme bands which include 3 forms of equal molecular weight, two of which consist of several isomers differing in their net charge. The MDH zymograms of the blue-green algae investigated can be used as “fingerprints”. The isoenzyme pattern of the MDHs of Anacystis nidulans makes its position in the order Chroococcales uncertain.

Résumé

Wäßrige Extrakte aus acht Cyanophyceenarten wurden einer Polyacrylamid-Discelektrophorese unterzogen und die erhaltenen Elektropherogramme auf Malat-Dehydrogenase (MDH)-Aktivität geprüft. Dabei ergab sich, daß unter unseren Versuchsbedingungen die MDH der getesteten Cyanophyceen in 5–8 Isoenzymbanden aufspaltet. Für Anabaena flos-aquae konnte gezeigt warden, daß sich die 8 Isoenzymbanden auf wenigstens 3 Molekulargewichts-Isomere zurückführen lassen, von denen zwei noch mehrere Ladungsisomere bilden. Die erhaltenen Zymogramme zeigen “fingerprint”-Charakter. was ihre mögliche Verwendbarkeit für die Chemotaxonomie der Cyanophyta nahelegt. Die Stellung von Anacystis nidulans innerhalb der CyanophyceenOrdnungen wird diskutiert.     

A comparison of the influence of iron on the growth and nitrate metabolism of Anabaena and Scenedesmus

A comparative study of growth and nitrate metabolism of Anabaena flos-aquae (Lyng.) Bréb. and Scenedesmus bijugatus var. seriatus Chodat investigated possible mechanisms for the iron-stimulated increases in growth specific for blue-green algae in mixed algal communities. Algae were separately grown in an morganic medium with varying concentrations of iron and nitrate to determine the effects on each organism. Iron was found to be a limiting nutrient for cultures of both Anabaena and Scenedesmus as determined by chlorophyll a concentrations and cell enumeration. Both iron and nitrate stimulated the specific activity of nitrate reductase, nitrite reductase, and glutamine synthetase in Anabaena. Iron enrichment did not increase the activity of the enzymes in Scenedesmus, but inhibited the activity of nitrate reductase and glutamine synthetase. The stimulation of growth by iron in cells grown under iron limiting conditions was associated with increased nitrate metabolism in Anabaena but not in Scenedesmus.     

RESPONSE OF ANABAENA FLOS-AQUAE (CYANOPHYTA) NITROGENASE ACTIVITY TO SUDDEN PHOSPHATE DEPRIVATION1

Cultures of Anabaena flos-aquae (Lyng.) Breb. Were used to determine changes in nitrogenase activity (acetylene reduction) after external concentrations of phosphorus were lowered. Two days following immersion in phosphorus-free medium, nitrogenase activity (NA) had doubled and required 8 days to return to time zero levels. Subsequent long-term experiments showed that concentrations of soluble reactive phosphorus (SRP) released from the algae transferred into the –P medium reached maximum levels by day 3 and returned to initial low values by days 7–10. NA was always highest during this SRP release-reassimilation phase but steadily decreased after reassimilation was complete. Day 56 NA was 5–14% of initial activity. The data support the hypothesis that heterocyst and vegetative cell ATP pools are discrete and suggest that the short-term effects of phosphorus removal as an aquatic restoration technique need further study.     

PRODUCTION OF PARALYTIC SHELLFISH TOXINS BY APHANIZOMENON SP. LMECYA 31 (CYANOBACTERIA)1

We examined intracellular and extracellular paralytic shellfish toxins (PST) in a strain of Aphanizomenon sp. (LMECYA31) isolated from a Portuguese freshwater reservoir throughout the growth cycle and under different conditions affected by temperature and nitrate and phosphate availability. PST concentrations and compositions were greatly influenced by cell density, growth stage, and temperature and nutrients conditions. On a per‐cell basis results showed (1) the enhancement of PST cell quota after the end of exponential growth phase in nutrient replete batch cultures, (2) the absence of a PST increment at late growth stages under phosphate limitation, (3) a rise in PST maximum cell quota under nitrate depletion, and (4) the enhancement of toxin production at higher temperatures. The relative proportion of the four toxins detected, neoSTX, dcSTX, STX and GTX5, also changed within and between culture settings. While growing under phosphate rich media cells produced mainly GTX5 and neoSTX, whereas under phosphate limitation the proportion of STX and dcSTX increased substantially with culture age. Large amounts of extracellular toxins were found in the culture medium, increasing during culture time. Extracellular toxin composition in each culture was fairly constant and always similar to the intracellular composition found at late stages of growth. This further supported other research that indicates that PSTs are released to the water through cell lysis, and a significant concentration of PST may be expected to remain in the water upon the collapse of a toxic bloom or after cells removal by water treatment.     

NOTE ESTABLISHMENT OF AXENIC CULTURES OF ANABAENA FLOS-AQUAE AND APHANOTHECE NIDULANS (CYANOBACTERIA) BY LYSOZYME TREATMENT

A new method using lysozyme for the production of axenic cultures of Anabaena flos-aquae De Brebisson and Aphanothece nidulans P. Richter was developed. Cyanobacterial growth was not inhibited at concentrations up to 1.2 g·L⁻¹ of lysozyme, whereas the growth of heterotrophic bacteria was suppressed. At concentrations up to 0.8 g·L⁻¹ of lysozyme, ampicillin caused a reduction of heterotrophic bacteria. The axenic cultures of these strains were acquired through a simple treatment using 1.0 g·L⁻¹ of lysozyme without ampicillin. These cyanobacteria resisted digestion by lysozyme at our experimental concentrations, whereas bacteria were digested selectively. This method of purification seems to be especially useful with cyanobacterial species that are sensitive to antibiotics or other germicidal agents.     

GENETIC VARIATION IN APHANIZOMENON (CYANOBACTERIA) COLONIES FROM THE BALTIC SEA AND NORTH AMERICA

Aphanizomenon Morren is an important member of the cyanobacterial community in the Baltic Sea, but studies of this genus have been hampered by the difficulty of growing it in laboratory culture. PCR amplification of DNA from colonies picked directly from water samples has circumvented this problem and made it possible to carry out an analysis of genetic diversity within the Baltic Sea and in two small North American lakes separated by just a few kilometers. The nucleotide sequence of the phycocyanin intergenic spacer and partial flanking coding regions of cpcB and cpcA was determined for 32 colonies of Aphanizomenon , 26 from the Baltic Sea, and 6 from the North American lakes. No variation was detected among the 26 Baltic Sea colonies, but two alleles, differing at 19 nucleotide positions (5.4%), were found in the North American lake colonies. Surprisingly, the two North American types were less closely related to each other than to the Baltic Sea genotype. The Baltic Sea Aphanizomenon is clearly distinct from A. flos-aquae at both the cpcB–cpcA and 16S rDNA loci, which lends phylogenetic support to their tentative separation based on ultrastructural analysis. We conclude that although there is significant genetic diversity in the genus Aphanizomenon , the population in the Baltic Sea is, in contrast to the Nodularia population, genetically homogeneous.     

Changes to the phytoplankton assemblage of Lake Kinneret after decades of a predictable, repetitive pattern

1. Phytoplankton abundance and species composition in Lake Kinneret, Israel, have been monitored at weekly or fortnightly intervals since 1969. This paper summarises the resulting 34‐year phytoplankton record with a focus on the last 13 years of new data, and reassesses an earlier conclusion that the lake phytoplankton shows remarkable stability despite a wide range of external pressures. 2. The Kinneret phytoplankton record can be split into two major periods. The first, from 1969 till 1993, was a period of distinct stability expressed by a typical annual pattern revolving around a spring bloom of the dinoflagellate Peridinium gatunense that repeated each year. The second period, starting around 1994 and ongoing, is characterised by the loss of the previously predictable annual pattern, with both ‘bloom years’ and ‘no‐bloom years’. 3. In the second period, deviations from the previous annual pattern include: the absence of the prevailing spring P. gatunense blooms in some years and increased variability in the magnitude of the bloom in others; intensification of winter Aulacoseira granulata blooms; higher summer phytoplankton biomass with replacement of mostly nanoplanktonic, palatable forms by less palatable forms; new appearance and establishment of toxin‐producing, nitrogen fixing cyanobacteria in summer; increase in the absolute biomass and percentage contribution of cyanobacteria to total biomass; and fungal epidemics attacking P. gatunense. 4. The 34‐year record serves to validate Schindler's (1987) assessment that phytoplankton species composition will respond to increased anthropogenic stress before bulk ecosystem parameters.     

水华鱼腥藻生长与光合作用对大气CO2浓度升高的响应

 为了探讨大气CO2浓度升高对水华藻类的影响,利用水华鱼腥藻(Anabena flos_aquae)作为实验材料,研究了大气CO2浓度加倍对其生长和光合作用的影响,结果显示大气CO2浓度升高导致水华鱼腥藻的生物量、光饱和光合速率、光合效率和光系统II的光化学效率（Fv/Fm）明显提高,但对暗呼吸速率和光饱和点没有明显影响。CO2加倍条件下藻细胞光合作用对无机碳的亲和力降低,表明其利用HCO-3的能力受到抑制。     

Effect of temperature and soluble reactive phosphorus on abundance of Aphanizomenon flos-aquae (Cyanophyceae)

Filament density of Aphanizomenon flos‐aquae (Lemmerm.) Ralfs, water temperature and soluble reactive phosphorus (SRP) were measured from April to August in 1993–1996 in Lake Barato, Hokkaido, Japan. In addition, growth characteristics and internal phosphorus (P) utilization of Aph. flos‐aquae were evaluated under P limitation at three temperatures (15, 20 and 25?C) to clarify the role of internal accumulated P for its growth in the incubation experiment. The filament density was highest in early July 1994, when SRP concentration had not yet decreased and the water temperature was high. These are important factors favoring an increase in abundance of this species in L. Barato. During batch culture, the time course of the stationary phase was shortest at 25?C and longest at 15?C; the cellular C:P molar ratio was 111 under P sufficiency and increased eight‐ to 12‐fold under P limitation. As the C:P ratio was significantly higher in the decreasing phase at 15?C, Aph. flos‐aquae may be more adaptable to Plimitation at 15?C than at 20?C and 25?C. However, the low temperatures did not favor the abundance of Aph. flos‐aquae in 1996. This indicates that the filament density of Aph. flos‐aquae decreases before it reaches the maximum value for some reason under P limitation in L. Barato.