Spectral Irradiance and Distribution of Pigments in a Highly Layered Marine Microbial Mat

The spectral irradiance from 400 to 1,100 nm was measured with depth in the intertidal sand mats at Great Sippewissett Salt Marsh, Mass. These mats contained at least four distinct layers, composed of cyanobacteria, purple sulfur bacteria containing bacteriochlorophyll a (Bchl a), purple sulfur bacteria containing Bchl b, and green sulfur bacteria. Spectral irradiance was measured directly by layering sections of mat on a cosine receptor. Irradiance was also approximated by using a calibrated fiber-optic tip. With the tip, irradiance measurements could be obtained at depth intervals less than 250 μm. The irradiance spectra were correlated qualitatively and quantitatively with the distribution of the diverse chlorophyll pigments in this mat and were compared with spectra recorded in plain sand lacking pigmented phototrophs. We found that the shorter wavelengths (400 to 550 nm) were strongly attenuated in the top 2 mm of the mat. The longer wavelengths (red and near infrared) penetrated to much greater depths, where they were attenuated by Bchl a, b, and c-containing anoxygenic phototrophic bacteria. The specific attenuation bands in the irradiance spectra correlated with the specific in vivo absorption bands of the Bchl-protein complexes in the bacteria. We concluded that the pigments in the phototrophs had a profound affect on the light environment within the mat. It seems likely that the diverse Bchl-protein complexes found in the anoxygenic phototrophs evolved in dense mat environments as a result of competition for light.     

Ecophysiology of stromatolitic microbial mats,Stocking Island,exuma cays,Bahamas

Intertidal stromatolites, covered by cyanobacterial mats, were recently discovered at Stocking Island, Exuma Cays, Bahamas. Ecophysiological responses (CO₂ fixation, N₂ fixation, and photoacclimation) of these cyanobacterial mats to experimental manipulations were examined to identify potential environmental variables controlling community structure and function. The mats exhibit horizontal zonation that shifts from soft to crusty to hard in a seaward direction. Cluster analysis of chemotaxonomic photopigments (chlorophylls and carotenoids) revealed that visually distinct mat types are composed of distinct phototrophic assemblages. Under reduced irradiance, diatoms within the mats photoacclimated by increasing accessory photopigments (diadinoxanthin, fucoxanthin, and chlorophyll c ₁ c ₂) and cyanobacteria reduced the photoprotective carotenoid echinenone. In a 4-day nutrient addition bioassay experiment, nitrate, phosphate, dissolved organic carbon, and trace metal enrichments did not enhance CO₂ fixation, but phosphate enrichments tripled N₂ fixation rates. The addition of DCMU increased N₂ fixation rates relative to nonamended light and dark rates, indicating light (photosystem I) enhanced nitrogenase activity. Soft mats appear to represent the early stages of colonization and stabilization of mat communities. Active growth following stabilization results in the formation of partially-lithified crusty mats, which eventually become highly-lithified and form hard mats. Collectively, our results suggest that Stocking Island stromatolitic mats have low growth rates and consequently exhibit slow responses to increased nutrient availability and changes in ambient irradiance. In general, intertidal stromatolitic mats at Stocking Island appear to exhibit low rates of CO₂ and N₂ fixation relative to nonlithifying temperate cyanobacteral mats. Although production is low, respiration is likewise low, leading to the suggestion that high production to respiration ratios (P:R) may be necessary for lithification of intertidal stromatolitic mats.     

Relations between electron transport rates determined by pulse amplitude modulated chlorophyll fluorescence and oxygen evolution in macroalgae under different light conditions

The relationship between O₂-based gross photosynthesis (GP) and in vivo chlorophyll fluorescence of Photosystem II-based electron transport rate (ETR) as well as the relationship between effective quantum yield of fluorescence (Φ_PSII) and quantum yield of oxygen evolution (Φ_{O_2}) were examined in the green algae Ulva rotundata and Ulva olivascens and the red alga Porphyra leucosticta collected from the field and incubated for 3 days at 100 μmol m⁻² s⁻¹ in nutrient enriched seawater. Maximal GP was twice as high in Ulva species than that measured in P. leucosticta. In all species ETR was saturated at much higher irradiance than GP. The initial slope of ETR versus absorbed irradiance was higher than that of GP versus absorbed irradiance. Only under absorbed irradiances below saturation or at values of GP <2 μmol O₂ m⁻² s⁻¹ a linear relationship was observed. In the linear phase, calculated O₂ evolved /ETR molar ratios were closed to the theoretical value of 0.25 in Ulva species. In P. leucosticta, the estimated GP was associated to the estimated ETR only at high irradiances. ETR was determined under white light, red light emitting by diodes and solar radiation. In Ulva species the maximal ETR was reached under red light and solar radiation whereas in P. leucosticta the maximal ETR was reached under white light and minimal under red light. These results are in agreement with the known action spectra for photosynthesis in these species. In the case of P. leucosticta, GP and ETR were additionally determined under saturating irradiance in algae pre-incubated for one week under white light at different irradiances and at white light (100 μmol m⁻² s⁻¹) enriched with far-red light. GP and growth rate increased at a growth irradiance of 500 μmol m⁻² s⁻¹ becoming photoinhibited at higher irradiances, while ETR increased when algae were exposed to the highest growth irradiance applied (2000 μmol m⁻² s⁻¹). The calculated O₂ evolved /ETR molar ratios were close to the theoretical value of 0.25 when algae were pre-incubated under 500–1000 μmol m⁻² s⁻¹. The enrichment by FR light provoked a decrease in both GP and ETR and an increase of nonphotochemical quenching although the irradiance of PAR was maintained at a constant level. In addition to C assimilation, other electron sinks, such as nitrogen assimilation, affected the GP–ETR relationship. The slopes of GP versus ETR or Φ_PSII versus Φ_{O_2} were lower in the algae with the highest N assimilation capacity, estimated as nitrate reductase activity and internal nitrogen contents, i.e., Ulva rotundata and Porphyra leucosticta, than that observed in U. olivascens. The possible mechanisms to explain this discrepancy between GP and ETR are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Arctic cyanobacteria and limnological properties of their environment: Bylot Island, Northwest Territories, Canada (73°N, 80°W)

Cyanobacteria were a major constituent of phototrophic communities in the lakes, ponds and streams of Bylot Island, in the Canadian high Arctic. The waters spanned a range of temperatures (1.8–16.8°C in late July), pH regimes (6.2–9.2) and conductivities (1.5–1700 μS cm⁻¹) but nutrient concentrations were consistently low (< 1 μg dissolved reactive P l⁻¹ at all sites; < 10 μg NO₃-N l⁻¹ at most sites). Picoplanktonic species (Synechococcus spp.) were often the numerical dominants in the plankton, and periphytic filamentous species (Oscillatoriaceae) commonly formed thick (5–50 mm) benthic mats. Bloom-forming species of cyanobacteria were either absent or poorly represented even in Chla-rich ponds. The total community biomass ranged from 0.1 to 29.8 μg Chla l⁻¹ in the plankton and from 1.1 to 34.8 μg Chla cm⁻² in the benthos. The in vivo absorbance characteristics of isolates from these environments indicated a genetically diverse range of species in each group of Arctic cyanobacteria. Growth versus irradiance relationships were determined for each of the isolates and similarly revealed large genetic differences (maximum growth rates from 0.17 to 0.61 day⁻¹), even between morphologically identical taxa. A comparison of nutrients, pigment concentrations and species composition underscores the strong similarities between freshwater ecosystems in the north and south polar zones. Received: 3 June 1996 / Accepted: 3 November 1996     

Energy Budget for the Cultured,Zooxanthellate Octocoral <Emphasis Type="Italic">Sinularia flexibilis</Emphasis>

The zooxanthellate octocoral Sinularia flexibilis is a producer of potential pharmaceutically important metabolites such as antimicrobial and cytotoxic substances. Controlled rearing of the coral, as an alternative for commercial exploitation of these compounds, requires the study of species-specific growth requirements. In this study, phototrophic vs. heterotrophic daily energy demands of S. flexibilis was investigated through light and Artemia feeding trials in the laboratory. Rate of photosynthetic oxygen by zooxanthellae in light (≈200 μmol quanta m⁻² s⁻¹) was measured for the coral colonies with and without feeding on Artemia nauplii. Respiratory oxygen was measured in the dark, again with and without Artemia nauplii. Photosynthesis–irradiance curve at light intensities of 0, 50, 100, 200, and 400 μmol quanta m⁻² s⁻¹ showed an increase in photosynthetic oxygen production up to a light intensity between 100 and 200 μmol quanta m⁻² s⁻¹. The photosynthesis to respiration ratio (P/R > 1) confirmed phototrophy of S. flexibilis. Both fed and non-fed colonies in the light showed high carbon contribution by zooxanthellae to animal (host) respiration values of 111–127%. Carbon energy equivalents allocated to the coral growth averaged 6–12% of total photosynthesis energy (mg C g ⁻ 1 buoyant weight day ⁻ 1) and about 0.02% of the total daily radiant energy. “Light utilization efficiency (ε)” estimated an average ε value of 75% 12 h ⁻ 1 for coral practical energetics. This study shows that besides a fundamental role of phototrophy vs. heterotrophy in daily energy budget of S. flexibilis, an efficient fraction of irradiance is converted to useable energy.     

Artificial Cyanobacterial Mats: Growth, Structure, and Vertical Zonation Patterns

Abstract The formation of cyanobacterial mats (originally induced by incubation of sediment cores in which metazoans and most other eukaryotes had been removed) was followed over approximately 2.6 years. The thickness of the mats increased at a rate of 2–3 mm per year because of accumulation of empty cyanobacterial sheaths and as a result of carbonate deposition; the fraction of living biomass remained relatively constant over at least 2 years, but there was a slow accumulation of nonliving organic C (≈ 1 mmol yr⁻¹). Biota composition (dominated by five types of filamentous cyanobacteria, unicellular cyanobacteria, diatoms, anoxygenic phototrophs, and heterotrophic bacteria) and vertical zonation patterns in the upper 2–3 mm of the mats were also almost constant over time. Using transmission electron microscopy and stereological analysis it was possible to quantify the vertical distribution of major groups of organisms. Received: 20 December 1999; Accepted: 10 June 2000; Online Publication: 28 August 2000     

Cyanobacterial mats from hot springs produce antimicrobial compounds and quorum-sensing inhibitors under natural conditions

Polar (water) and non-polar (ethyl acetate) extracts from the cyanobacterial layer (top 1–3 mm) of four hot spring microbial mats in the Sultanate of Oman were tested for their antibacterial, antidiatom and quorum-sensing inhibitory activities under natural conditions. The chemical composition of the active extracts was analysed using gas chromatography–mass spectrometry (GC-MS). Cyanobacteria within these mats were identified by direct microscopy while the total bacterial community composition was compared using automated ribosomal intergenic spacer analysis (ARISA). Only the extracts from Bowshar and Nakhl mats showed antibacterial properties against Bacillus sp., Micrococcus luteus, Shigella sonnei, Salmonella enterica and Klebsiella pneumoniae. All tested extracts inhibited the growth of the benthic diatom Amphora coffeaeformis. Extracts from Bowshar, Rustaq and Nakhl inhibited quorum-sensing of the reporter strains Chromobacterium violaceum CV017 and Agrobacterium tumefaciens NTL4. The highest bioactivity was recorded for ethyl acetate extracts from Nakhl mats, which had the lowest number of operational taxonomic units (OTUs). Using GC-MS, 74 chemical compounds were obtained, however with different distribution among the four mat extracts (similarity < 43%). Various cyanobacteria, belonging mainly to Chroococcus, Phormidium, Leptolyngbya, Spirulina and Lyngbya were detected in the different mats, and each mat had its unique bacterial community, as confirmed by ARISA profiles. We conclude that antimicrobial and quorum-sensing inhibitory compounds can be produced by hot spring mat microorganisms under natural conditions and the differences in these compounds could be attributed to the differences in the mats’ bacterial composition as well as the physical–chemical conditions of the springs.     

Nitrogen fixation by the benthic freshwater cyanobacterium Lyngbya wollei

The ability of the benthic cyanobacterium Lyngbya wollei to fix nitrogen was studied using field samples and axenic cultures. L. wollei was collected and isolated from Lake Okeechobee, Florida, where it forms extensive mats. Rates of acetylene reduction up to 39.1 nmol mg dry wt⁻¹ h⁻¹ were observed for field samples. The maximum observed rate of acetylene reduction in axenic laboratory cultures was 200 nmol mg dry wt⁻¹ h⁻¹. Aerobic conditions limited nitrogen fixation activity, but dark/light cycles promoted the development of activity. Reduced oxygen levels appeared to be required for the development of significant levels of nitrogenase activity. The level of irradiance also had a significant impact on the level of activity. The potential significance of nitrogen fixation to Lyngbya production is discussed.     

Oxygenic Photosynthesis and Respiratory Activity in Microbial Mats of the Ebro Delta, Spain, by Oxygen Exchange Method

Photosynthetic and respiratory activities at low light intensities (300 μE m⁻² s⁻¹) in the microbial mats of the Ebro Delta were measured by the oxygen exchange method in the laboratory. The response to H₂S concentration, a significant factor in the dynamics of that ecosystem, was assessed. Total photosynthesis reached 23.78–28.17 μg O₂ cm⁻² h⁻¹. Photosynthetic activity was not significantly different at the two temperatures tested. Respiratory activity reached a consumption of 6.95–8.56 μg O₂ cm⁻² h⁻¹ at 25°C and 11.42–11.70 μg O₂ cm⁻² h⁻¹ at 35°C. The Q₁₀ value for respiration was 1.37–1.64. Oxygen production in Microcoleus chthonoplastes, the most abundant cyanobacterium in those microbial mats, was highly resistant to sulfide inhibition. Concentrations less than 0.02 mM sulfide did not affect the rate of photosynthesis. Concentrations up to 0.1 mM sulfide caused different degrees of partially reversible inhibition, with a maximum of 67% at 0.78 mM sulfide. Primary production (g C assimilated/m²/year) in those microbial mats was also assessed and compared with data from other ecosystems. Received: 24 October 1997 / Accepted: 18 December 1997     

Oxygen and nutrient dynamics of the upside down jellyfish (Cassiopea sp.) and its influence on benthic nutrient exchanges and primary production

The oxygen and nutrient dynamics of the zooxanthellate, upside down jellyfish (Cassiopea sp.), were determined both in situ and during laboratory incubations under controlled light conditions. In the laboratory, Cassiopea exhibited a typical Photosynthesis–Irradiance (P–I) curve with photosynthesis increasing linearly with irradiance, until saturation was reached at an irradiance of ~400 μE m⁻² s⁻¹, with photosynthetic compensation (photosynthesis = respiration) being achieved at an irradiance of ~50 μE m⁻² s⁻¹. Under saturating irradiation, gross photosynthesis attained a rate of almost 3.5 mmol O₂ kg WW⁻¹ h⁻¹, whereas the dark respiration rate averaged 0.6 mmol O₂ kg WW⁻¹ h⁻¹. Based upon a period of saturating irradiance of 9 h, the ratio of daily gross photosynthesis to daily respiration was 2.04. Thus, photosynthetic carbon fixation was not only sufficient to meet the carbon demand of respiration, but also to potentially support a growth rate of ~3% per day. During dark incubations Cassiopea was a relatively minor source of inorganic N and P, with the high proportion of NO _X (nitrate + nitrite) produced indicating that the jellyfish were colonised by nitrifying bacteria. Whereas, under saturating irradiance the jellyfish assimilated ammonium, NO _X and phosphate from the bathing water. However, the quantities of inorganic nitrogen assimilated were small by comparison to carbon fixation rates and the jellyfish would need to exploit other sources of nitrogen, such as ingested zooplankton, in order to maintain balanced growth. During in situ incubations the presence of Cassiopea had major effects on benthic oxygen and nutrient dynamics, with jellyfish occupied patches of sediment having 3.6-fold higher oxygen consumption and 4.5-fold higher ammonium regeneration rates than adjacent patches of bare sediment under dark conditions. In contrast at saturating irradiance, jellyfish enhanced benthic photosynthetic oxygen production almost 100-fold compared to the sediment alone and created a small sink for inorganic nutrients, whereas unoccupied sediment patches were sources of inorganic nutrients to the water column. Overall, Cassiopea greatly enhanced the spatial and temporal heterogeneity of benthic fluxes and processes by creating “hotspots” of high activities which switched between being sources or sinks for oxygen and nutrients over diurnal irradiance cycles, as the metabolism of the jellyfish swapped between heterotrophy and net autotrophy.     

Confocal Laser Scanning Microscopy Image Analysis for Cyanobacterial Biomass Determined at Microscale Level in Different Microbial Mats

We recently published a new method based on determining cyanobacterial biomass by confocal laser scanning microscopy image analysis (CLSM-IA) (Solé et al., Ultramicrosc 107:669–673, 2007). CLSM-IA allows biomass calculation for microorganisms of a small size, since the limit of the technique’s resolution is that generated by a voxel, the smallest unit of a three-dimensional digital image, equivalent to 1.183 × 10⁻³ mgC/cm³ of sediment. This method is especially suitable for the quantitative analysis of a large number of CLSM images generated from benthic sediments in which complex populations of cyanobacteria are abundant, such as microbial mats. In order to validate the new CLSM approach, mats with varying structural characteristics were studied. We have grouped them into three types: Microcoleus mats (laminated), sandy mats (nonlaminated and composed of well-sorted quartz sands), and oil-polluted mats. In this work, we applied CLSM-IA in natural [the Ebro delta and Sant Jordi colony (Spain), Salins-de-Giraud and Etang de Berre (France), and Orkney Islands (Scotland)] and artificial [mesocosms (Israel)] microbial mats. A total of 4,103 confocal images were obtained in order to determine total and individual cyanobacteria biomass profiles, at microscale level. The data presented in this paper show the efficacy of the method, as it can be applied to highly diverse mat samples.     

Photosynthetic acclimation to photon irradiance and its relation to chlorophyll fluorescence and carbon assimilation in the halotolerant green alga Dunaliella viridis

This work describes the long-term acclimation of the halotolerant microalga Dunaliella viridis to different photon irradiance, ranging from darkness to 1500 μmol m⁻² s⁻¹. In order to assess the effects of long-term photoinhibition, changes in oxygen production rate, pigment composition, xanthophyll cycle and in vivo chlorophyll fluorescence using the saturating pulse method were measured. Growth rate was maximal at intermediate irradiance (250 and 700 μmol m⁻² s⁻¹). The increase in growth irradiance from 700 to 1500 μmol m⁻² s⁻¹ did not lead to further significant changes in pigment composition or EPS, indicating saturation in the pigment response to high light. Changes in Photosystem II optimum quantum yield (F_v/F_m) evidenced photoinhibition at 700 and especially at 1500 μmol m⁻² s⁻¹. The relation between photosynthetic electron flow rate and photosyntetic O₂ evolution was linear for cultures in darkness shifting to curvilinear as growth irradiance increased, suggesting the interference of the energy dissipation processes in oxygen evolution. Carbon assimilation efficiencies were studied in relation to changes in growth rate, internal carbon and nitrogen composition, and organic carbon released to the external medium. All illuminated cultures showed a high capability to maintain a C:N ratio between 6 and 7. The percentage of organic carbon released to the external medium increased to its maximum under high irradiance (1500 μmol m⁻² s⁻¹). These results suggest that the release of organic carbon could act as a secondary dissipation process when the xanthophyll cycle is saturated. This revised version was published online in June 2006 with corrections to the Cover Date.     

An in situ study of photosynthetic oxygen exchange and electron transport rate in the marine macroalga Ulva lactuca (Chlorophyta)

Direct comparisons between photosynthetic O₂ evolution rate and electron transport rate (ETR) were made in situ over 24 h using the benthic macroalga Ulva lactuca (Chlorophyta), growing and measured at a depth of 1.8 m, where the midday irradiance rose to 400–600 μmol photons m⁻² s⁻¹. O₂ exchange was measured with a 5-chamber data-logging apparatus and ETR with a submersible pulse amplitude modulated (PAM) fluorometer (Diving-PAM). Steady-state quantum yield ((F_m′−F_t)/F_m′) decreased from 0.7 during the morning to 0.45 at midday, followed by some recovery in the late afternoon. At low to medium irradiances (0–300 μmol photons m⁻² s⁻¹), there was a significant correlation between O₂ evolution and ETR, but at higher irradiances, ETR continued to increase steadily, while O₂ evolution tended towards an asymptote. However at high irradiance levels (600–1200 μmol photons m⁻² s⁻¹) ETR was significantly lowered. Two methods of measuring ETR, based on either diel ambient light levels and fluorescence yields or rapid light curves, gave similar results at low to moderate irradiance levels. Nutrient enrichment (increases in [NO₃ ⁻], [NH₄ ⁺] and [HPO₄ ^2-] of 5- to 15-fold over ambient concentrations) resulted in an increase, within hours, in photosynthetic rates measured by both ETR and O₂ evolution techniques. At low irradiances, approximately 6.5 to 8.2 electrons passed through PS II during the evolution of one molecule of O₂, i.e., up to twice the theoretical minimum number of four. However, in nutrient-enriched treatments this ratio dropped to 5.1. The results indicate that PAM fluorescence can be used as a good indication of the photosynthetic rate only at low to medium irradiances. This revised version was published online in June 2006 with corrections to the Cover Date.     

Dinitrogen-Fixing Cyanobacteria in Microbial Mats of Two Shallow Coral Reef Ecosystems

Dinitrogen-fixing organisms in cyanobacterial mats were studied in two shallow coral reef ecosystems: La Reunion Island, southwestern Indian Ocean, Sesoko (Okinawa) Island, and northwestern Pacific Ocean. Rapidly expanding benthic miniblooms, frequently dominated by a single cyanobacterial taxon, were identified by microscopy and molecular tools. In addition, nitrogenase activity by these blooms was measured in situ. Dinitrogen fixation and its contribution to mat primary production were calculated using ¹⁵N₂ and ¹³C methods. Dinitrogen-fixing cyanobacteria from mats in La Reunion and Sesoko showed few differences in taxonomic composition. Anabaena sp. among heterocystous and Hydrocoleum majus and Symploca hydnoides among nonheterocystous cyanobacteria occurred in microbial mats of both sites. Oscillatoria bonnemaisonii and Leptolyngbya spp. occurred only in La Reunion, whereas Hydrocoleum coccineum dominated in Sesoko. Other mats dominated by Hydrocoleum lyngbyaceum, Phormidium laysanense, and Trichocoleus tenerrimus occurred at lower frequencies. The 24-h nitrogenase activity, as measured by acetylene reduction, varied between 11 and 324 nmoles C₂H₂ reduced μg⁻¹ Chl a. The highest values were achieved by heterocystous Anabaena sp. performed mostly during the day. Highest values for nonheterocystous cyanobacteria were achieved by H. coccineum mostly during the night. Daily nitrogen fixation varied from nine (Leptolyngbya) to 238 nmoles N₂ μg⁻¹ Chl day⁻¹ (H. coccineum). Primary production rates ranged from 1,321 (S. hydnoides) to 9,933 nmoles C μg⁻¹ Chl day⁻¹ (H. coccineum). Dinitrogen fixation satisfied between 5% and 21% of the nitrogen required for primary production.     

Periphyton nitrogenase activity as an indicator of wetland eutrophication: spatial patterns and response to phosphorus dosing in a northern Everglades ecosystem

The use of periphyton nitrogenase activity (biological N₂ fixation) as an indicator of wetland P impact was assessed using patterns of nutrient content (C, N, P, Ca, Mg, K, Fe, and Mn) and acetylene reduction (AR) in floating cyanobacterial periphyton mat (metaphyton) communities of a P-enriched portion of the Florida Everglades, USA (Water Conservation Area-2A, WCA-2A). Spatial patterns of nutrients indicate the enrichment of floating mat periphyton N, P, Fe, and K, and the reduction of Mn and TN:TP in enriched marsh areas. In highly enriched areas, floating mat periphyton AR was approximately threefold greater than that in less enriched, interior marsh zones. Multiple regression models indicated AR dependence on P in eutrophic WCA-2A areas while the AR of more interior marsh periphyton mats was more closely related to tissue levels of Ca and Fe. Nitrogenase activity of floating mat periphyton from P-loaded mesocosms revealed a significant enhancement of N₂ fixation in samples receiving approximately 2–3 mg P m⁻² of cumulative P dosing or with biomass TP content of 100–300 mg kg⁻¹. At P contents above the optimum, mat periphyton AR was suppressed possibly as a result of changes in species composition or increased levels of NH₄⁺. After 3 years of dosing, consistently high AR occurred only at low rates of P enrichment (0.4–0.8 g P m⁻² yr⁻¹), and the patterns appeared to be seasonal. These findings agree with the hypothesis that P availability is a key determinant of nitrogenase activity in aquatic systems, and thus, may support the use of periphyton nitrogenase to indicate P impacts in P-limited systems. These results also demonstrate the potential existence of a P threshhold for biogeochemical alteration of periphyton mat function in the Everglades, and that cumulative loading of limiting nutrients (i.e., P), rather than instantaneous concentrations, should be considered when evaluating nutrient criteria.     

Redox Changes of Iron Caused by Erosion,Resuspension and Sedimentation in Littoral Sediment of a Freshwater Lake

Depth profiles of oxygen concentration and the redox status of acid-extractable iron were measured in littoral sediment cores of Lake Constance after mechanical removal of surface sediment, mixing, and re-deposition. In undisturbed sediment cores, oxygen penetrated down to 2.9±0.4 mm or 4.6±0.4 mm depth, respectively, after 12 h of incubation in the dark or light; causing a net diffusive flux of 108±20 nmol cm⁻² h⁻¹ oxygen into or 152±35 nmol cm⁻² h⁻¹ out of the sediment. The uppermost 20 mm layer of the undisturbed sediment cores contained 10.2± 0.7 μmol cm⁻³ ferrous and 3.8±1.1 μmol cm⁻³ ferric iron. After erosion, the oxic–anoxic interface in the newly exposed sediment was shifted to about 2 mm depth within 30 min, causing an oxygen flow into the sediment. During the following 12 h, oxygen penetrated deeper into the sediment, and in the light oxygen was produced photosynthetically. Ferrous iron was largely oxidized within two days after erosion. The oxidation rates were higher in oxic than in anoxic sediment layers, and decreased with time. This oxidation process took the longer and was confined closer to the surface the more reduced the exposed sediment had been before. Resuspension of eroded sediment in aerated lake water did not cause a significant oxidation or reduction of iron. After re-deposition, the oxic–anoxic interface in the re-sedimented material shifted to about 2 mm depth within 30 min, causing an oxygen flow into the sediment. During the following 12 h, the oxygen penetration depth and the oxygen flow rate into the re-deposited sediment did not change any further, and no oxygen was produced in the light. Ferric iron was reduced during the first day after re-deposition, and partly re-oxidized during the second day. The extent of reduction was stronger and the extent of oxidation weaker the more reduced the resuspended sediment was before. Oxic conditions in the sediment surface were established faster and ferrous iron was oxidized to a larger extent after erosion of sediment than after resuspension and sedimentation.     

Complementary limiting factors of astaxanthin synthesis during photoautotrophic induction of <Emphasis Type="Italic">Haematococcus pluvialis</Emphasis>: C/N ratio and light intensity

We investigated the effect of carbon/nitrogen (C/N) ratio on astaxanthin synthesis in Haematococcus pluvialis during photoautotrophic induction by continuous input of both CO₂–air mixture and intense light. When H. pluvialis was induced by constant irradiance induction at 200 μmol photon m⁻² s⁻¹, there was a positive correlation with astaxanthin content and C/N ratio, which was similar to the case for heterotrophic induction. Lower C/N ratios did not retard Haematococcus encystment, but did increase culture biomass, resulting in a decrease in astaxanthin production because of light limitation. However, induction using variable irradiance showed that reduction of astaxanthin production at low C/N ratios was successfully overcome by simply increasing the light intensity from 200 to 300 μmol photon m⁻² s⁻¹ to overcome the light limitation. This resulted in a greatly enhanced astaxanthin synthesis in proportion to cell density in cultures with low C/N ratios. Our results indicate that light intensity is more critical than C/N ratio in astaxanthin production by H. pluvialis during photoautotrophic induction.     

Limnological characteristics of the freshwater ecosystems of Byers Peninsula, Livingston Island, in maritime Antarctica

A limnological survey of 15 lakes and 6 streams was carried out on Byers Peninsula (Livingston Island, South Shetland Islands, Antarctica) during austral summer 2001–2002. Most of the surface waters had low conductivities (20–105 μS cm⁻¹) and nutrients (total phosphorus 0.01–0.24 μM), but some coastal lakes were enriched by nutrient inputs from seal colonies and marine inputs. Plankton communities in the lakes contained picocyanobacteria (10²–10⁴ cells ml⁻¹), diatoms, chrysophytes and chlorophytes, and a large fraction of the total biomass was bacterioplankton. Zooplankton communities were dominated by Boeckella poppei and Branchinecta gainii; the benthic cladoceran Macrothrix ciliata was also recorded, for the first time in Antarctica. The chironomids Belgica antarctica and Parochlus steinenii, and the oligochaete Lumbricillus sp., occurred in stream and lake benthos. The phytobenthos included cyanobacterial mats, epilithic diatoms and the aquatic moss Drepanocladus longifolius. These observations underscore the limnological richness of this seasonally ice-free region in maritime Antarctica and its value as a long-term reference site for monitoring environmental change.     

Light-induced quenching of chlorophyll fluorescence at 77 K in leaves,chloroplasts and Photosystem II particles

The light-induced chlorophyll (Chl) fluorescence decline at 77 K was investigated in segments of leaves, isolated thylakoids or Photosystem (PS) II particles. The intensity of chlorophyll fluorescence declines by about 40% upon 16 min of irradiation with 1000 μmol m⁻² s⁻¹ of white light. The decline follows biphasic kinetics, which can be fitted by two exponentials with amplitudes of approximately 20 and 22% and decay times of 0.42 and 4.6 min, respectively. The decline is stable at 77 K, however, it is reversed by warming of samples up to 270 K. This proves that the decline is caused by quenching of fluorescence and not by pigment photodegradation. The quantum yield for the induction of the fluorescence decline is by four to five orders lower than the quantum yield of Q_A reduction. Fluorescence quenching is only slightly affected by addition of ferricyanide or dithionite which are known to prevent or stimulate the light-induced accumulation of reduced pheophytin (Pheo). The normalised spectrum of the fluorescence quenching has two maxima at 685 and 695 nm for PS II emission and a plateau for PS I emission showing that the major quenching occurs within PS II. ‘Light-minus-dark’ difference absorbance spectra in the blue spectral region show an electrochromic shift for all samples. No absorbance change indicating Chl oxidation or Pheo reduction is observed in the blue (410–600 nm) and near infrared (730–900 nm) spectral regions. Absorbance change in the red spectral region shows a broad-band decrease at approximately 680 nm for thylakoids or two narrow bands at 677 and 670–672 nm for PS II particles, likely resulting also from electrochromism. These absorbance changes follow the slow component of the fluorescence decline. No absorbance changes corresponding to the fast component are found between 410 and 900 nm. This proves that the two components of the fluorescence decline reflect the formation of two different quenchers. The slow component of the light-induced fluorescence decline at 77 K is related to charge accumulation on a non-pigment molecule of the PS II complex. This revised version was published online in June 2006 with corrections to the Cover Date.