Phycoerythrin and photosynthesis of the pelagic blue-green alga Trichodesmium thiebautii in the waters of Kuroshio,Japan

The photosynthetic function of phycoerythrin was investigated in the alga Trichodesmium thiebautii from the waters of Kuroshio, Japan. The spectroscopic characteristics of the in vivo and isolated T. thiebautii phycoerythrin pigments are identical, and have 3 absorption bands at 495, 547 and 562 nm. Light at the wavelengths corresponding to each absorption band of phycoerythrin is equally efficient in T. thiebautii photosynthesis, indicating that phycoerythrin is active in trapping light energy for photosynthesis. In the natural habitat, phycoerythrin is considered to be the main photosynthetic pigment in T. thiebautii photosynthesis.     

Ultrastructure of the gonad and gametogenesis in the eastern oyster,Crassostrea virginica. II. Testis and spermatogenesis

The pelagic harpacticoid copepod, Macrosetella gracilis (A. Scott), is found in association with colonies of the nitrogen-fixing (diazotrophic), bloomforming cyanobacterium Trichodesmium spp. in tropical and subtropical waters. M. gracilis is one of the few direct grazers of these often toxic cyanobacteria. Experiments investigating NH ₊ ⁴ regeneration by M. gracilis were conducted in the Caribbean in September 1992 and the Coral Sea, Australia in November 1994. Rates of M. gracilis ingestion of Trichodesmium thiebautii labelled with ¹⁵N₂ measured in the eastern Caribbean indicated that M. gracilis could consume 33 to 45% of total T. thiebautii colony N d^-1 and >100% of new N fixed d^-1. We also measured the release of NH ₊ ⁴ by M. gracilis feeding on T. thiebautii, as well as by non-feeding copepods, using ¹⁵N isotope dilution methods. In non-feeding copepods, rates of NH ₊ ⁴ release increased as numbers of copepods were increased as both copepod numbers and food availability increased. In the presence of T. thiebautii colonies, M. gracilis had an average rate of NH ₊ ⁴ regeneration of 7.7±1.5 nmol N copepod^-1 h^-1 (±SE), which was significantly higher than when food was absent (1.9±0.7 nmol N copepod^-1 h^-1). Rates of M. gracilis excretion were relatively high based on excretion: ingestion ratios, which could be due to having a high-N food source readily available, to sloppy-feeding effects, or as a response to toxins in the cyanobacterium. Incubations of M. gracilis with and without T. erythraeum resulted in significant increases in [NH ₊ ⁴ ] as a function of copepod density only. Ammonium leakage from the cyanobacterium and/or microheterotroph associates was relatively low. M. gracilis, through excretion and possible mechanical breakage of cells while grazing, appears to provide a direct link between atmospherically derived new nitrogen and regenerated NH ₊ ⁴ in the oligotrophic systems where Trichodesmium spp. are abundant.     

Aerobic nitrogenase activity measured as acetylene reduction in the marine non-heterocystous cyanobacterium Trichodesmium spp. grown under artificial conditions

Aerobic nitrogenase activity in the marine non-heterocystous cyanobacterium Trichodesmium spp. NIBB 1067, isolated off the Izu Peninsula, Japan in 1983 and grown under artificial conditions, was assayed by the acetylene reduction method. This strain exhibited acetylene reduction activity under aerobic conditions when cells had been grown in the medium free of combined nitrogen. Activity was markedly enhanced by light, and dependent on the growth phase being higher during the exponential growth phase and lower during the late linear and stationary growth phases. Since typical colony formation occurred during the last growth phase, the present results contradict the idea that N₂-fixation depends on colony formation. The photosynthesis inhibitor DCMU at 10^-6 M inhibited light-dependent acetylene reduction completely. Acetylene reduction by Trichodesmium spp. was tolerant of O₂ as strongly as that in the heterocystous cyanobacteria. Even at a partial pressure of oxygen (p_{O 2}) of 3 atm, the activity still remained as high as half of the maximum. It was almost under anaerobic conditions. Maximum activity was obtained at pO₂ of ca. 0.1 atm.     

Nitrogen fixation (acetylene reduction) associated with the living coral Acropora variabilis

To estimate N₂-fixation, acetylene reduction assays were carried out on portions of the branches of the coral Acropora variabilis from the west coast of Malaysia. In some experiments, a sub-surface incubation apparatus was employed that was designed to keep the coral fragments near to their natural depth of occurrence. Other shipboard experiments used metabolic inhibitors to investigate the class of organism reducing acetylene. Stumps of coral gave the highest rates of activity, probably attributable to loosely associated cyanophytes. Coral tips also reduced acetylene at relatively high rates; reduction was enhanced in light by increased CO₂ concentration and decreased O₂ tensions indicative of photosynthetic bacteria. Algal material was not obvious on the tip surfaces and so the active organism was probably more integral to the coral structure than it was in the stumps. Maximum rates of acetylene reduction measured translated to 2.5 mg N₂ fixed per outcrop per day.     

Seasonal variations in nitrogen-fixation (acetylene reduction) and sulphate-reduction rates in the rhizosphere of Zostera noltii: nitrogen fixation by sulphate-reducing bacteria

Nitrogen-fixation (acetylene reduction) rates were measured over an annual cycle in meadows of the seagrass Zostera noltii Hornem in the Bassin d'Arcachon, south-west France, between March 1994 and February 1995, using both slurry and whole-core techniques. Measured rates using the slurry technique consistently overestimated those determined on whole cores, probably due to the release of labile organic carbon sources as a result of root damage during preparation of the slurries. Thus, the whole-core technique may provide a more accurate estimate of in situ activity, since disturbance of physicochemical gradients of oxygen, sulphide, nutrients and the relationship between the plant roots and the rhizosphere microflora is minimised. Rates measured by the whole-core method were 1.8- to 4-fold greater (dependent upon season) in the light than those measured during dark incubations, indicating that organic carbon diffusing from the plant roots during photosynthesis was an important factor in regulating nitrogen fixation in the rhizosphere. Additions of sodium molybdate, a specific inhibitor of sulphate-reducing bacteria (SRB) inhibited acetylene-reduction activity by >80% as measured by both the slurry and whole-core techniques throughout the year, inferring that SRB were the dominant component of the nitrogen-fixing microflora. A mutualistic relationship between Z. noltii and nitrogen-fixing SRB in the rhizosphere, based on the exchange of organic carbon and fixed nitrogen is proposed. Acetylene- and sulphate-reduction rates showed distinct summer peaks which correlated with a reduced availability of ammonium in the sediment and the annual growth cycle of Z. noltii in the basin. Overall, these data indicate that acetylene reduction (nitrogen fixation) activity in the rhizosphere of Z. noltii was regulated both by the availability of organic carbon from the plant roots and maintenance of a low NH ₄ ⁺ concentration in the vicinity of the plant roots due to efficient assimilation of NH ₄ ⁺ by Z. noltii during the growth season. Nitrogenfixation rates determined from acetylene-reduction rates measured using the whole-core technique ranged from 0.1 to 7.3 mg N m^-2d^-1, depending on season, and were calculated to contribute between 0.4 and 1.1 g N m^-2yr^-1, or 6.3 to 12% of the annual fixed nitrogen requirement of Z. noltii.     

Hydrogen production by nitrogenase as a potential crop rotation benefit

Both climate change and the adverse effects of chemical use on human and environmental health are recognized as serious issues of global concern. Nowhere is this more apparent than in the agricultural sector where release of greenhouse gases such as carbon dioxide, nitrous oxide and methane continues to be problematic and where use of nitrogen fertilizer is responsible for negative impacts on both human populations and ecosystems. The manipulation of biological nitrogen fixation (BNF) could help alleviate part of the difficulty by decreasing the need for nitrogen fertilizers, which require huge quantities of fossil fuel to produce and contribute to the release of nitrous oxide from soil as well as being responsible for the contamination of drinking water systems and natural habitats. BNF is performed by a variety of microorganisms. One of the most studied examples is the BNF carried out by rhizobial bacteria in symbiosis with their plant hosts such as pea and soybean. Hydrogen gas is an energy-rich, obligate by-product of BNF. Legume symbioses with rhizobia lacking hydrogenase enzymes (which can recycle hydrogen) have traditionally been viewed as energetically inefficient. However, recent studies suggest hydrogen release to soil may be beneficial, increasing soil carbon sequestration and promoting growth of hydrogen-oxidizing bacteria beneficial to plant growth; the alleged superiority of symbiotic performance in rhizobia possessing functional hydrogenases (HUP⁺) over those rhizobia without functional hydrogenases (HUP⁻) has also not been conclusively shown. The structure of the iron-molybdenum cofactor or FeMo-co of nitrogenase (the active site of the enzyme) has been elucidated through X-ray crystallography but the mechanism of nitrogen fixation remains unknown. However, studies of effects of hydrogen production on BNF have revealed potential candidate intermediates involved in the nitrogenase reaction pathway and have also shown the role of hydrogen as a competitive inhibitor of N₂, with hydrogen now considered to be the primary regulator of the nitrogenase electron allocation coefficient. The regulation of oxygen levels within legume root nodules is also being investigated; nitrogen fixation is energetically expensive, requiring a plentiful oxygen supply but too high an oxygen concentration can irreversibly damage nitrogenase, so some regulation is needed. There is evidence from gas diffusion studies suggesting the presence of a diffusion barrier in nodules; leghaemoglobin is another potential O₂ regulator. Possible functions of hydrogenases include hydrogen recycling, protection of nitrogenase from damaging O₂ levels and prevention of inhibitory H₂ accumulation; there is evidence for H₂ recycling only in studies where H₂ uptake has been strongly coupled to ATP production and where this is not the case, it is believed that the hydrogenase acts as an O₂ scavenger, lowering O₂ concentrations. The distribution of hydrogenases in temperate legumes has been found to be narrow and root and shoot grafting experiments suggest the host plant may exert some influence on the expression of hydrogenase (HUP) genes in rhizobia that possess them. Many still believe that HUP⁺ rhizobia are superior in performance to HUP⁻ species; to this end, many attempts to increase the relative efficiency of nitrogenase through the introduction of HUP genes into the plasmids or chromosomes of HUP⁻ rhizobia have been carried out and some have met with success but many other studies have not revealed an increase in symbiotic performance after successful insertion of HUP genes so the role of HUP in increasing parameters such as N₂ fixation and plant yield is still unclear. One advantage of the hydrogen production innate to BNF is that the H₂ evolved can be used to measure N₂ fixation using new open-flow gas chamber techniques seen as superior to the traditional acetylene reduction assay (ARA) conducted in closed chambers, although H₂ cannot be used for field studies yet as the ARA can. However, the ARA is now believed to be unreliable in field studies and it is recommended that other measures such as dry weight, yield and total nitrogen content are more accurate, especially in determining real food production, particularly in the developing nations. Another potential benefit of H₂ release from root nodules is that it stays in the soil and has been found to be consumed by H₂-oxidizing bacteria, many of which show plant growth–promoting properties such as the inhibition of ethylene biosynthesis in the host plant, leading to root elongation and increased plant growth; they may well be promising as biofertilizers if they can be successfully developed into seed inoculants for non-leguminous crop species, decreasing the need for chemical fertilizers. It has been suggested that rhizobia can produce nitrous oxide through denitrification but this has never been shown; it is possible that hydrogen release may provide more ideal conditions for denitrifying, free-living bacteria and so increase production of nitrous oxide that way and this issue will require more study. However, it seems unlikely that a natural system would release nitrous oxide to the same degree that chemical fertilizers have been shown to do.     

Nitrogen fixation associated with the cyanobacterial mat of a marine laminated microbial ecosystem

The nitrogenase activity in the cyanobacterial mat of a laminated microbial ecosystem was investigated by the acetylene reduction method. Measurements under several conditions such as light and dark, aerobic and anaerobic and by inhibiting photosystem II by 10^-5 M DCMU showed the nitrogenase activity to be light stimulated and to some degree inhibited by oxygen. An appreciable amount of activity was also present under complete aerobic conditions. We estimated 8 to 15 kg N fixed per hectare per year for that part of the intertidal flat supporting growth of cyanobacteria. By measuring a vertical sediment profile, nitrogenase was shown to be associated with the cyanobacterial mat. Diurnal measurements of nitrogenase showed two activity peaks, one at sunrise and one at sunset. Following population dynamics in the cyanobacterial mat showed Microcoleus sp., Oscillatoria spp., Spirulina sp., Gloeocapsa sp. and sometimes Merismopedia sp. to be present. During four years of observations we never found any heterocystous cyanobacteria. Non-heterocystous cyanobacteria apparently play an important role in nitrogen fixation in this marine intertidal environment.     

Cultures of the pelagic cyanophytes Trichodesmium erythraeum and T. thiebautii in synthetic medium

Trials for determination of culture conditions for the marine cyanophytes of Trichodesmium erythraeum and T. thiebautii were made with use of a synthetic medium. The Aquil medium, either with or without combined nitrogen, brought about stable growth of the two strains, T. erythraeum and T. thiebautii. However, they failed to grow in an ASP₇ medium. The failure was found to be due to the toxic effect of Tris-aminomethane, the pH-buffer in this medium. Two important chemical conditions for the stable growth of Trichodesmium spp. were revealed. (1) Stable growth was supported by Ca²⁺ at high concentrations; in a concentration lower than 0.9 mM, cell-lysis promptly occurred, while the cells could grow without cell-lysis at Ca²⁺ concentrations higher than 7.5 mM even at a salinity as low as 19 S. Ca²⁺ is probably essential for the osmotic regulation in this organism. (2) Phosphate-toxicity at high concentrations was at least partly due to heavy metal(s) contaminating the reagent of inorganic phosphate. After treatment with a Chelex-100 column, phosphate concentration could be increased up to four times the previous concentrations without toxicity.     

Grazing effects on nitrogen fixation in coral reef algal turfs

This study addressed whether grazing by the sea urchin Diadema antillarum influenced rates of nitrogen fixation by algal turf communities on Caribbean coral reefs. Because the turfs were nitrogen-limited, we also assessed whether newly-fixed nitrogen was important for supporting net primary productivity by the turfs. We measured acetylene reduction in turfs grown in treatments excluding or including D. antillarum in the presence of other herbivores at 3 m water depth on Tague Bay forereef, St. Croix, U.S. Virgin Islands. These were the first measurements of acetylene reduction on coral reefs under quasi-natural conditions of high water-flow and photosynthetic oxygen generation. Rates of acetylene reduction under these conditions were as high as any measured previously in coral reef communities (mean 7.6 nmol C₂H₄ cm⁻² h⁻¹). Algal turfs grazed by D. antillarum and other herbivores had chlorophyll-specific acetylene reduction rates up to three times higher than when D. antillarum was excluded. High rates of nitrogen fixation by the turfs were sufficient to meet <2% of the nitrogen required to support net chlorophyll-specific primary productivity over 24 h. Grazer-mediated increases in nitrogen fixation do not appear responsible for a parallel enhancement of net primary productivity. Algal turfs at this site must be dependent primarily on external sources of nitrogen. Received: 1 July 1997 / Accepted: 5 September 1997     

Nitrogen fixation by blue-green algae associated with the siphonous green seaweed Codium decorticatum: effects on ammonium uptake

Nitrogen fixation (acetylene reduction) at rates of up to 1.2 g N₂ g dry wt^-1 h^-1 was measured for the siphonous green seaweed Codium decorticatum. No nitrogenase activity was detected in C. isthmocladum. The nitrogenase activity was light sensitive and was inhibited by the addition of DCMU and triphenyl tetrazolium chloride. Additions of glucose did not stimulate nitrogen fixation. Blue-green algae (Calothrix sp., Anabaena sp., and Phormidium sp.) were implicated as the organisms responsible for the nitrogenase activity. They occurred in a reduced microzone within the C. decorticatum thallus where nitrogen fixation was optimized. Nitrogen fixation did not affect the kinetic constants for ammonium uptake in C. decorticatum (K_s=12.0 M, V_max=13.4 mol NH₃ g dry wt^-1 h^-1) determined using the perturbation method. Nevertheless, C. decorticatum thalli which fixed nitrogen had internal dissolved nitrogen concentrations which were over 1.4 times higher than in non-fixing thalli. This suggests that if C. decorticatum does derive part of its nitrogen requirement from the blue-green algae which it harbors, the transfer does not involve competition between this process and the uptake of ambient ammonium.     

Acetylene reduction (nitrogen fixation) in a Delaware,USA salt marsh

Acetylene reduction (nitrogen fixation) was measured in several vegetational areas in a Delaware, USA salt marsh. Samples were collected for 1 yr and the results showed a seasonally variable pattern of acetylene reduction at all stations. Peak rates were generally recorded during the later summer and early fall (September–October). The seasonality was influenced mainly, although not exclusively, by the soil temperature. In addition, samples collected in short Spartina alterniflora stands exhibited rates which were up to 20-fold higher than those found in samples from tall S. alterniflora stands. Over 50% of the total yearly ethylene production occurred from mid-August until the beginning of December at the tall and short S. alterniflora stations. Maximum activity occurred at 5 cm depth for all stations. Surface activity accounted for only 3–4% of the total measured in the top 20 cm. Addition of glucose or mannitol resulted in considerable increases in activity, thus suggesting that heterotrophic acetylene reduction is carbon and/or energy limited. The results obtained in this study indicate that the measured rates are only potential rates and that considerable caution must be used in extrapolating from acetylene reduction rates to nitrogen fixation rates in situ.     

Experimental ecology of the temperate scleractinian coral Astrangia danae I. Partition of respiration,photosynthesis and calcification between host and symbionts

Calcification, photosynthesis and respiration of the scleractinian coral Astrangia danae were calculated from the changes in total alkalinity, pH, calculated total CO₂, and oxygen concentration produced by colonies incubated in glass jars. A correction for changes in ammonia, nitrate and nitrite was taken into account and the method evaluated. The fluxes of oxygen and CO₂ were highly correlated (r=0.99). The statistical error of alkalinity determinations was less than 10% of the changes observed in the slowest calcifying samples. Metabolism of polyparium alone was estimated by difference after removal of tissue and reincubation of bare corallum. Zooxanthellae concentration in the polyps was obtained from cell counts made on homogenates of polyp tissue. The calculated photosynthetic rate of the zooxanthellae in vivo was 25 mol O₂ (10⁸ cell)^-1 h^-1 at a light intensity of 120 Ein m^-2 s^-1. In corals having 0.5x10⁹ zooxanthellae/dm² of colony area up to 8% of the total photosynthesis was attributed to the corallum microcosm. Polyp respiration, photosynthesis, and CaCO₃ uptake rates were all much higher than rates previously reported from A. danae, apparently because in these experiments the organisms were better fed. This increased photosynthesis in turn enhanced calcification still further. The symbiosis therefore appears to provide a growth advantage even to fed corals, under the conditions of these experiments.     

Nitrogen fixation in the rhizosphere of marine angiosperms

High rates of acetylene reduction were observed in systems containing excised rhizomes of the Caribbean marine angiosperms Thalassia testudinum, Syringodium filiforme and Diplanthera wrightii, and the temperate marine angiosperm Zostera marina. For 4 plant and plant-sediment systems the ratio of acetylene reduced/N₂ fixed varied from 2.6 to 4.6. For T. testudinum the estimated rates of nitrogen fixation are in agreement with estimated requirements of the plant for nitrogen. For a typical T. testudinum stand, N₂ fixation is estimated to be 100 to 500 kg N/hectare per year. Numbers of N₂-fixing bacteria in the rhizosphere sediments were roughly 50 to 300 times more abundant than those in the nonrhizosphere sediments, and in both types of sediments were of the same orders as the estimated numbers of heterotrophic aerobes.Canadian IBP Contribution No. 137.     

Effects of feeding frequency and symbiosis with zooxanthellae on nitrogen metabolism and respiration of the coral Astrangia danae

Colonies of the temperate coral Astrangia danae occur naturally with and without zooxanthellae. Basal nitrogen excretion rates of nonsymbiotic colonies increased with increasing feeding frequency [average excretion rate was 635 ng-at N (mg-at tissue-N)^-1 h^-1]. Reduced excretion rates of symbiotic colonies were attributed to N uptake by the zooxanthellae. Nitrogen uptake rates of the zooxanthellae averaged 8 ng-at N (10⁶ cells)^-1 h^-1 in the dark and 21 ng-at N (10⁶ cells)^-1 h^-1 at 200 Ein m^-2 s^-1. At these rates the zooxanthellae could provide 54% of the daily basal N requirement of the coral if all of the recycled N was translocated. Basal respiration rates were 172 nmol O₂ cm^-2 h^-1 for starved colonies and 447 nmol O₂ cm^-2 h^-1 for colonies fed three times per week. There were no significant differences between respiration rates of symbiotic and nonsymbiotic colonies. N excretion and respiration rates of fed (symbiotic and nonsymbiotic) colonies increased greatly soon after feeding. N absorption efficiencies decreased with increasing feeding frequency. A N mass balance, constructed for hypothetical situations of nonsymbiotic and symbiotic (3×10⁶ zooxanthellae cm^-2) colonies, starved and fed 15 g-at N cm^-2wk^-1, showed that the presence of symbionts could double the N growth rate of feeding colonies, and reduce the turnover-time of starved ones, but could not provide all of the N requirements of starved colonies. Rates of secondary production, estimated from rates of photosynthesis and respiration were similar to those estimated for reef corals.     

Oxygen-radical-mediated toxic effects of the red tide flagellate Chattonella marina on Vibrio alginolyticus

Toxic mechanisms of the red tide flagellate, Chattonella marina, collected in 1985 from Kagoshima Bay, Japan, were studied at the subcellular level. C. marina was found to reduce ferricytochrome c at a rate related to the concentration of plankton cells. Ca. 50% of the cytochrome c reduction was inhibited by the addition of 100 U superoxide dismutase ml^-1. These results suggest that a part of the cytochrome c reduction was caused by a superoxide anion which was extracellulary released from C. marina. Moreover, a small amount of hydrogen peroxide was detected in the C. marina suspension using the fluorescence spectrophotometric assay method. The identity of the hydrogen peroxide was confirmed by its reaction with 500 U catalase ml^-1. It is thus proposed that C. marina produces harmful active oxygen radicals and therefore exhibits a toxic effect on surrounding living organisms. In agreement with these results, C. marina strongly inhibited the proliferation of marine bacteria, Vibrio alginolyticus, in a plankton/bacteria co-culture system. The growth inhibition of bacteria caused by C. marina was related to the density and the metabolic potential of C. marina. Ruptured plankton showed no toxic effect on the bacteria. Furthermore, the toxic effect of C. marina on V. alginolyticus was completely suppressed by the addition of catalase and superoxide dismutase. In addition to these radical-scavenging enzymes, a chemical scavenger, sodium benzoate, also had a protective effect. These results suggest that oxygen radicals are important in the toxic action of C. marina.     

Feeding biology and ecological impact of an introduced nudibranch,Tritonia plebeia,New England,USA

The nudibranchTritonia plebeia (Johnston) was first observed in New England in 1983, on vertical rock walls at 7 m depth off Nahant, Massachusetts. This northern European species preys exclusively on the soft coralAlcyonium digitatum (Linneaus) in its natural habitat. At Nahant, it preyed primarily on the closely relatedAlcyonium siderium Verrill. Laboratory studies indicated that it could locate its prey by distance chemoreception and by visual orientation towards tall dark surfaces which could help it find the vertical walls, overhangs, and boulder sides where the soft corals occur. Field studies showed thatT. plebeia fed primarily on colony bases, causing extensive damage and whole colony mortality. The most important endemic predator onA. siderium, Coryphella verrucosa (Sars), preyed preferentially on hydroids, but would graze polyps off the top portions ofA. sederium colonies, causing little permanent damage to the colony, during the winter months when hydroids were scarce. AlthoughC. verrucosa occasionally behaved agonistically towardT. plebeia, there was no indication in laboratory or field studies that either nudibranch had an effect on the other's foraging through interference competition. Extensive predation byT. plebeia caused the disappearance ofA. siderium at two sites (Outer and Inner Shag Rocks) and a sharp reduction at a third site (Inner East Point). The higher mortalities at the Shag Rocks sites most likely occurred because of a simultaneous urchin (Strogylocentrotus droebachiensis) population expansion. As space among aggregates ofA. siderium opened up due toT. plebeia predation, urchins were able to forage on the vertical walls and scrape off remaining colonies. At a fourth site, Halfway Rock, whereT. plebeia were seldom present,A. siderium colonies also suffered high mortalities. This increae in mortality began nearly a year before urchin populations increased, and during a summer of abnormally high water temperatures at Halfway Rock. The high temperatures, followed by urchin predation on remaining colonies could account for the disappearance of allA. siderium colonies at this site.T. plebeia disappeared at all sites by summer 1986 andA. siderium populations have since stabilized, but community-level changes at all sites whereA. siderium were removed have persisted.     

Daily rhythm of O2-evolution in the cyanobacterium Trichodesmium thiebautii under natural and constant conditions

The rate of oxygen evolution by the tropical marine cyanobacterium Trichodesmium thiebautii was recorded at different times during the day in samples collected in 1992 from the Bahama Islands and the NE Caribbean Sea. This cyanobacterium is unique in that it is the only non-heterocystous diazotroph capable of N₂-fixation in daylight. Oxygen evolution was measured under conditions of natural day/night (LD, N=50), constant light (LL, N=14), and constant dark (DD, N=2×14). Photosynthesis vs intensity (P-I) relationships were calculated at various times of day, and the following parameters were used for further evaluation: photosynthesic capacity (P _max, 66 to 91 mg O₂ mg chl a ^-1 h^-1), initial slope of the P-I curve (, 0.23 to 0.27), dark respiration (R, 12 to 27 mg O₂ mg chl a ^-1 h^-1), and the intensity at which O₂ consumption is compensated by O₂ production (I _c, 78 to 160 Em^-2 s^-1). All means showed large standard deviations (for some parameters more than 200%). In some cases, these variations could be explained with a sinusoidal 24-h time course, but only the compensation point showed a significant daily variation (p0.001) in both LD and DD. The fact that the time course of I _c typical for natural conditions remains rhythmic under constant dark conditions strongly suggests a circadian regulation. Few circadian rhythms have been observed in prokaryotes, and this appears to be the first observation of such a rhythm in a cyanobacterium which fixes N₂ in daytime.     

Inorganic nitrogen metabolism inUlva rigida illuminated with blue light

Inorganic nitrogen metabolism in blue light was studied for the green algaUlva rigida C. Agardh collected in the south of Spain (Punta Carnero, Algeciras) in the winter of 1987. NH₄ ⁺ has been reported to inhibit NO₃ ^- uptake; however,U. rigida showed a net NO₃ ^- uptake even when the NH₄ ⁺ concentration of the external medium was three or four times greater than the concentration of NO₃ ^-. NO₃ ^- uptake rates were similar in both darkness and in blue light of various photon fluence rates (PFR) ranging from 17 to 160 mol m^-2 s^-1. Since NO₃ ^- uptake is an active mechanism involving the consumption of ATP, respiratory metabolism can provide enough ATP to maintain the energetic requirement of NO₃ ^- transport even in darkness. In contrast, NO₃ ^- reduction inU. rigida was highly dependent on the net photosynthetic rate. After 7 h in blue light, intracellular NO₃ ^- concentrations ([NO₃ ^-] _i ) were higher in specimens exposed to intensities below the light compensation point (LCP) than in those incubated at a PFR above the LCP. When PFR is below the light compensation point, NO₃ ^- reduction is low, probably because all the NADH produced by the cells is oxidized in the respiratory chain in order to produce ATP to maintain a steady NO₃ ^- transport rate. The total nitrogen (TN) and carbon (TC) contents decreased from darkness to 33 mol m^-2 s^-1 in blue light. In this range, catabolic processes prevailed over anabolic ones. In contrast, increases in TN and TC contents were observed above the light compensation point. The C : N ratio increased with light intensity, reaching a stable value of 17 at 78 mol m^-2 s^-1 in blue light. Intracellular NO₃ ^- concentration and NO₃ ^- reduction appear to be directly controlled by light intensity. This external control of [NO₃ ^-]_i and the small capacity ofU. rigida to retain incorporated NO₃ ^-, NO₂ ^- and NH₄ ⁺ ions may explain its nitrophilic character.     

Interplay of host morphology and symbiont microhabitat in coral aggregations

The Belizean reef coral Agaricia tenuifolia Dana forms aggregations in which rows of thin, upright blades line up behind each other. On average, the spacing between blades increases with depth and hence with decreasing ambient irradiance. We designed and built a small, inexpensive light meter and used it to quantify the effect of branch spacing on light levels within colonies at varying distances from branch tips. Concurrently, we measured photosynthetic pigment concentrations and population densities of symbiotic dinoflagellates (zooxanthellae) extracted from coral branches of colonies with tight (≤3 cm) vs wide (≥6 cm) branch spacing, collected at 15 to 17 m and from colonies with tight branch spacing collected at 1 to 2 m. Light levels decreased significantly with tighter branch spacing and with distance from the branch tips. Total cellular pigment concentrations (chlorophylls a, c ₂ and peridinin) as well as chlorophyll a:c ₂ and chlorophyll a: peridinin ratios all increased significantly with distance from the branch tip, indicating very localized differences in photoacclimation within individual branches. Zooxanthellae from colonies with widely-spaced branches displayed significantly lower chlorophyll a:c ₂ and chlorophyll a:peridinin ratios, and were present at significantly higher population densities than those from colonies with tightly-spaced branches collected at the same depth (15 m). Tightly-spaced colonies collected from shallow environments (1 to 2 m) displayed pigment ratios similar to those from widely-spaced colonies from deeper water (15 m), but maintained zooxanthellae populations at levels similar to those in tightly-branched colonies from deeper water. Thus, variation in colony morphology (branch spacing and distance from branch tip) can affect symbiont physiology in a manner comparable to an increase of over 15 m of water depth. These results show that a host's morphology can strongly determine the microhabitat of its symbionts over very small spatial scales, and that zooxanthellae can in turn display steep gradients in concordance with these altered physical conditions. Received: 12 June 1997 / Accepted: 24 June 1997     

Electrocardiogram of a marine fish,Pagrus major,exposed to red tide plankton,Chattonella marina

Examinations of the electrocardiogram of Pagrus major exposed to Chattonella marina, a planktonic organism causing red tide, were made to determine the physiological effects on fish. The heart rate decreased as a result of the extention in the interval between T and P waves. The decrease in heart rate with the extended intervals between T and P waves was also recognized in the condition of decreased dissolved oxygen. Also, the decrease in heart rate of the fish exposed to the red tide occurred while the fish was struggling. This reduction seems to be caused by the strong tension of the vagal nerve. Upon exposure to C. marina at high cell concentration, the heart beat at a very low frequency after 30 min. The very low heart rate is expected to limit seriously the oxygen uptake by the gill, because the cardiac output is probably very low in this situation.