Photolimitation and photoinhibition as factors determining optimal dilution rate to produce eicosapentaenoic acid from cultures of the microalga Isochrysis galbana

Eicosapentaenoic acid (EPA) productivity from continuous cultures of the marine microalga Isochrysis galbana was studied, taking into account the irradiance on the reactor surface, that is, the photolimitation/photoinhibition regime to which the cells are exposed. Experiments were conducted under a wide variety of operating conditions. The dilution rate ranged from 0.005 h⁻¹ to 0.040 h⁻¹ at five external irradiances (820, 1620, 2050, 2450 and 3270 μmol photons m⁻² s⁻¹) covering photolimited to photoinhibited growth. Under these conditions, the specific growth rate (μ) was found to be the main factor influencing EPA content (ranging from 2.35% to 5.23% dryweight) and productivity (up to 0.88 mg l⁻¹ h⁻¹). The fatty acid content was not significantly affected by the external irradiance, but was influenced by the state of growth of the microalga, depending on whether the light regime was photolimiting or photoinhibiting. It might be suggested that light should no longer be considered an isolated factor affecting EPA synthesis, but an indirect influence through the photolimitation/photoinhibition regime and growth rate. At a given dilution rate, EPA content and biomass concentration are lower under photoinhibiting external irradiances than those corresponding to photolimiting conditions, and consequently EPA productivity decays. Since the effect of photoinhibition is less marked at high biomass concentration, a strategy to optimize EPA productivity from microalgal cultures could consist of reducing the dilution rate when the external irradiance increases above the phoinhibition threshold. Received: 16 January 1998 / Revised revision: 27 March 1998 / Accepted: 27 March 1998     

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.     

Growth rates of phytoplankton under fluctuating light

• 1 The effect of light fluctuations on the growth rates of four species of freshwater phytoplankton was investigated. Experimental light regimes included constant irradiance and fluctuations of a step function form, with equal proportion of high (maximum of 240 µmol photons m^‐2 s^‐1) and low light (minimum of 5 µmol photons m^‐2 s^‐1) (or dark) in a period. Fluctuations of 1, 8 and 24‐h periods were imposed over several average irradiances (25, 50, 100 and 120 µmol photons m^‐2 s^‐1).
• 2 Growth rate responses to fluctuations were species‐specific and depended on both the average irradiance and the period of fluctuations. Fluctuations at low average irradiances slightly increased growth rate of the diatom Nitzschia sp. and depressed growth of the cyanobacterium Phormidium luridum and the green alga Sphaerocystis schroeteri compared to a constant irradiance.
• 3 Fluctuations at higher average irradiance did not have a significant effect on the growth rates of Nitzschia sp. and Sphaerocystis schroeteri (fluctuations around saturating irradiances) and slightly increased the growth rates of the cyanobacteria Anabaena flos‐aquae and Phormidium luridum (when irradiance fluctuated between limiting and inhibiting levels).
• 4 In general, the effect of fluctuations tended to be greater when irradiance fluctuated between limiting and saturating or inhibiting levels of a species growth‐irradiance curve compared to fluctuations within a single region of the curve.
• 5 The growth rates of species under fluctuating light could not always be predicted from their growth‐irradiance curves obtained under constant irradiance. When fluctuations occur between limiting and saturating or inhibiting irradiances for the alga and when the period of fluctuations is long (greater than 8 h), steady‐state growth‐irradiance curves may be insufficient to predict growth rates adequately. Consequently, additional data on physiological acclimation, such as changes in photosynthetic parameters, may be required for predictions under non‐constant light supply in comparison to constant conditions.

     

Eco-physiological responses of nitrogen-fixing cyanobacteria to light

The eco-physiological responses of three nitrogen-fixing cyanobacteria (N-fixing cyanobacteria), Aphanizomenon gracile, Anabaena minderi, and Ana. torques-reginae, to light were assessed under nutrient saturation. The N-fixing cyanobacteria were isolated into monocultures from a natural bloom in a shallow colored lake and their growth irradiance parameters and pigment composition were assessed. The different ecological traits related to light use (μ_max, α, I _k) suggest that these N-fixing cyanobacteria are well adapted to low light conditions at sufficient nutrients, yet interspecific differences were observed. Aphanizomenon gracile and Anabaena minderi had high relative growth rates at low irradiances (ca. 70% of those in high light), low half saturation constant for light-limited growth (I _k < 9.09 μmol photon m⁻² s⁻¹) and high efficiency (α < 0.11 day⁻¹ μmol photon⁻¹ m² s). Conversely, Ana. torques-reginae showed poorer light competitiveness: low relative growth rates at low irradiances (ca. 40% of those in high light), low α (0.009 day⁻¹ μmol photon⁻¹ m² s) and higher I _k (35.5 μmol photon m⁻² s⁻¹). Final densities in Aphanizomenon gracile and Anabaena minderi reached bloom densities at irradiances above 30 μmol photon m⁻² s⁻¹ with different hierarchy depending on irradiance, whereas Ana. torques-reginae never achieved bloom densities. All species had very low densities at irradiances ≤17 μmol photon m⁻² s⁻¹, thus no N-fixing blooms would be expected at these irradiances. Also, under prolonged darkness and at lowest irradiance (0 and 3 μmol photon m⁻² s⁻¹) akinetes were degraded, suggesting that in ecosystems with permanently dark sediments, the prevalence of N-fixing cyanobacteria should not be favored. All species displayed peaks of phycocyanin, but no phycoeritrin, probably due to the prevailing red light in the ecosystem from which they were isolated.     

Decrease of phosphoribulokinase activity by antisense RNA in transgenic tobacco: definition of the light environment under which phosphoribulokinase is not in large excess

 To test the hypothesis that the contribution of phosphoribulokinase (PRK) to the control of photosynthesis changes depending on the light environment of the plant, the response of transgenic tobacco (Nicotiana tabacum L.) transformed with antisense PRK constructs to irradiance was determined. In plants grown under low irradiance (330 μmol m⁻² s⁻¹) steady-state photosynthesis was limited in plants with decreased PRK activity upon exposure to higher irradiance, with a control coefficient of PRK for CO₂ assimilation of 0.25 at and above 800 μmol m⁻² s⁻¹. The flux control coefficient of PRK for steady-state CO₂ assimilation was zero, however, at all irradiances in plant material grown at 800 μmol m⁻² s⁻¹ and in plants grown in a glasshouse during mid-summer (alternating shade and sun 300–1600 μmol m⁻² s⁻¹). To explain these differences between plants grown under low and high irradiances, Calvin cycle enzyme activities and metabolite content were determined. Activities of PRK and other non-equilibrium Calvin cycle enzymes fructose-1,6-bisphosphatase, sedoheptulose-1,7-bisphosphatase and ribulose-1,5-bisphosphate carboxylase-oxygenase were twofold higher in plants grown at 800 μmol m⁻² s⁻¹ or in the glasshouse than in plants grown at 330 μmol m⁻² s⁻¹. Activities of equilibrium enzymes transketolase, aldolase, ribulose-5-phosphate epimerase and isomerase were very similar under all growth irradiances. The flux control coefficient of 0.25 in plants grown at 330 μmol m⁻² s⁻¹ can be explained because low ribulose-5-phosphate content in combination with low PRK activity limits the synthesis of ribulose-1,5-bisphosphate. This limitation is overcome in high-light-grown plants because of the large relative increase in activities of sedoheptulose-1,7-bisphosphatase and fructose-1,6-bisphosphatase under these conditions, which facilitates the synthesis of larger amounts of ribulose-5-phosphate. This potential limitation will have maintained evolutionary selection pressure for high concentrations of PRK within the chloroplast. Received: 15 November 1999 / Accepted: 27 January 2000     

Leaf photosynthesis, plant growth and nitrogen allocation in rice under different irradiances

The photosynthetic rates and various components of photosynthesis including ribulose-1,5-bisphosphate carboxylase (Rubisco; EC 4.1.1.39), chlorophyll (Chl), cytochrome (Cyt) f, and coupling factor 1 (CF₁) contents, and sucrose-phosphate synthase (SPS; EC 2.4.1.14) activity were examined in young, fully expanded leaves of rice (Oryza sativa L.) grown hydroponically under two irradiances, namely, 1000 and 350 μmol quanta · m⁻² · s⁻¹, at three N concentrations. The light-saturated rate of photosynthesis measured at 1800 μmol · m⁻² · s⁻¹ was almost the same for a given leaf N content irrespective of growth irradiance. Similarly, Rubisco content and SPS activity were not different for the same leaf N content between irradiance treatments. In contrast, Chl content was significantly greater in the plants grown at 350 μmol · m⁻² · s⁻¹, whereas Cyt f and CF₁ contents tended to be slightly smaller. However, these changes were not substantial, as shown by the fact that the light-limited rate of photosynthesis measured at 350 μmol · m⁻² · s⁻¹ was the same or only a little higher in the plants grown at 350 μmol · m⁻² · s⁻¹ and that CO₂-saturated photosynthesis did not differ between irradiance treatments. These results indicate that growth-irradiance-dependent changes in N partitioning in a leaf were far from optimal with respect to N-use efficiency of photosynthesis. In spite of the difference in growth irradiance, the relative growth rate of the whole plant did not differ between the treatments because there was an increase in the leaf area ratio in the low-irradiance-grown plants. This increase was associated with the preferential N-investment in leaf blades and the extremely low accumulation of starch and sucrose in leaf blades and sheaths, allowing a more efficient use of the fixed carbon. Thus, morphogenic responses at the whole-plant level may be more important for plants as an adaptation strategy to light environments than a response of N partitioning at the level of a single leaf. Received: 23 February 1997 / Accepted: 8 May 1997     

Acclimation of the summer annual species, Lolium temulentum, to CO2 enrichment

Lolium temulentum L. Ba 3081 was grown hydroponically in air (350 μmol mol⁻¹ CO₂) and elevated CO₂ (700 μmol mol⁻¹ CO₂) at two irradiances (150 and 500 μmol m⁻² s⁻¹) for 35 days at which point the plants were harvested. Elevated CO₂ did not modify relative growth rate or biomass at either irradiance. Foliar carbon-to-nitrogen ratios were decreased at elevated CO₂ and plants had a greater number of shorter tillers, particularly at the lower growth irradiance. Both light-limited and light-saturated rates of photosynthesis were stimulated. The amount of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) protein was increased at elevated CO₂, but maximum extractable Rubisco activities were not significantly increased. A pronounced decrease in the Rubisco activation state was found with CO₂ enrichment, particularly at the higher growth irradiance. Elevated-CO₂-induced changes in leaf carbohydrate composition were small in comparison to those caused by changes in irradiance. No CO₂-dependent effects on fructan biosynthesis were observed. Leaf respiration rates were increased by 68% in plants grown with CO₂ enrichment and low light. We conclude that high CO₂ will only result in increased biomass if total light input favourably increases the photosynthesis-to-respiration ratio. At low irradiances, biomass is more limited by increased rates of respiration than by CO₂-induced enhancement of photosynthesis. Received: 23 February 1999 / Accepted: 15 June 1999     

Seasonal variation of phytoplankton community structure and nitrogen uptake regime in the Indian Sector of the Southern Ocean

This study investigates the dynamics of phytoplankton communities and nitrogen uptake in the Indian sector of the Southern Ocean during spring and summer. The study area is oligotrophic (Chl a stocks <50 mg m⁻²); nevertheless, a large spatial variation of phytoplankton biomass and community structure was observed. During both seasons the phytoplankton community in the seasonal ice zone showed higher biomasses and was mainly composed of large diatom cells. However, in the permanently open ocean zone the community had low biomass and was chiefly composed of nano- and picoflagellates. In the polar front zone, although biomass was higher, the community structure was similar to the open ocean zone. The results suggest that the variation in phytoplankton community structure on a larger scale resonates with gradients in water column stability and nutrient distribution. However, significant changes in biomass and nutrient stocks but little change in community structure were observed. Absolute nitrogen uptake rates were generally low, but their seasonal variations were highly significant. During spring the communities displayed high specific nitrate uptake (mean rate = 0.0048 h⁻¹), and diatoms (in the seasonal ice zone) as well as nano- and picoflagellates (in the permanently open ocean zone and polar front zone) were mainly based on new production (mean ƒ-ratio = 0.69). The transition to summer was accompanied by a significant reduction in nitrate uptake rate (0.0048 h⁻¹ → 0.0011 h⁻¹) and a shift from predominantly new to regenerated production (ƒ-ratio 0.69 → 0.39). Ammonium played a major role in the seasonal dynamics of phytoplankton nutrition. The results emphasize that, despite a large contrast in community structure, the seasonal dynamics of the nitrogen uptake regime and phytoplankton community structure in all three subsystems were similar. Additionally, this study supports our previous conclusion that the seasonal shift in nitrogen uptake regime can occur with, as well as without, marked changes in community structure. Received: 2 December 1997 / Accepted: 20 April 1998     

Evaluation of the different levels of variability in the underwater light field of a shallow estuary

The underwater light climate of a shallow estuary located at the southern coast of the Baltic Sea has been investigated, with special emphasis on the spectral irradiance composition and on short-term irradiance fluctuations caused by vertical mixing and wave focussing. The inherent optical properties of the water body were dominated by phytoplankton pigment absorption in the long-wavelength range and by coloured, dissolved organic matter (cDOM) absorption in the wavelength range <500 nm, including ultraviolet-A (UV-A) and ultraviolet-B radiation (UV-B). Pronounced particulate scattering combined with the absorption values to give very high attenuation coefficients, especially for the shorter wavelengths of UV-B radiation. Photosynthetically active radiation (PAR) was found to be reduced to 1% of the surface value within 0.8 m in the inner, hypertrophic end of the estuary and within 1.9 m in the outer, eutrophic parts of this system, with corresponding 1% penetration depths for UV-B of 0.13 and 0.31 m. During late winter and early spring, the period when reduced atmospheric ozone concentrations and enhanced UV-B have been reported over northern Europe, the irradiance levels in the water column were greatly reduced, due to strong attenuation by ice cover and overlying snow. cDOM concentration of the water was also found to remain at a high level during these periods, and indeed throughout the year, thus reducing the exposure of organisms to UV-R and PAR still further. A complex irradiance regime was found in this system, with irregular and high amplitude fluctuations caused by wind-induced vertical mixing and wave focussing being superimposed upon the solar-angle-dependent seasonal and daily cycles. The methods used to quantify the short-term fluctuations are described, and their relevance to phytoplankton physiology is discussed. The wave-focussing effect is unique to the aquatic environment, and measurements showed that average subsurface irradiances could be increased by up to 5 times for periods lasting for <1 s. The highest irradiances recorded during wave-focussing events reached over 9,000 μmol photons m^–2 s^–1. Received in revised form: 7 April 2000 Electronic Publication     

Irradiance-dependent regulation of gravitropism by red light in protonemata of the moss Ceratodon purpureus

Apical cells of protonemata of the moss Ceratodon purpureus (Hedw.) Brid. are negatively gravitropic in the dark and positively phototropic in red light. Various fluence rates of unilateral red light were tested to determine whether both tropisms operate simultaneously. At irradiances ≥140 nmol m⁻² s⁻¹ no gravitropism could be detected and phototropism predominated, despite the presence of amyloplast sedimentation. Gravitropism occurred at irradiances lower than 140 nmol m⁻² s⁻¹ with most cells oriented above the horizontal but not upright. At these low fluence rates, phototropism was indistinct at 1 g but apparent in microgravity, indicating that gravitropism and phototropism compete at 1 g. The frequency of protonemata that were negatively phototropic varied with the fluence rate and the duration of illumination, as well as with the position of the apical cell before illumination. These data show that the fluence rate of red light regulates whether gravitropism is allowed or completely repressed, and that it influences the polarity of phototropism and the extent to which apical cells are aligned in the light path. Received: 19 January 1999 / Accepted: 19 March 1999     

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.     

The role of platelet ice microalgae in seeding phytoplankton blooms in Terra Nova Bay (Ross Sea,Antarctica): a mesocosm experiment

The aim of this study was to assess the role of platelet ice microalgal communities in seeding pelagic blooms. Nutrient dynamics, microalgal biomass, photosynthetic parameters, cell densities and species succession were studied in two mesocosm experiments, designed to simulate the transition of microalgal communities from platelet ice habitat to pelagic conditions. The microalgal assemblages were dominated by diatoms, 70% of which were benthic species such as Amphiprora kufferathii, Nitzschia stellata, and Berkeleya adeliensis. Photoacclimation of benthic species was inadequate also at relatively low irradiances. Exceptional growth capacity at different light levels was observed for pelagic species such as Fragilariopsis cylindrus and Chaetoceros spp. which may be important in seeding blooms at ice breakup. Fragilariopsis cylindrus showed high growth rates both at 65 and 10% of incident light and in nutrient replete as well as in nutrient depleted conditions. Five days after inoculation, phytoplankton biomass increased and nutrient concentrations decreased in both light conditions. Nutrient uptake rates were up to 9.10 μmol L⁻¹ d⁻¹ of TIN in the high light tank and 6.18 μmol L⁻¹ d⁻¹ in the low light tank and nutrient depletion in the high light tank occurred 3 days prior to depletion in the low light tank. At nutrient depletion, biomass concentrations were similar in both tanks, 30 and 34 μg Chla L⁻¹. This article belongs to a special topic: Five articles on Sea-ice communities in Terra Nova Bay (Ross Sea), coordinated by L. Guglielmo and V. Saggiomo, appear in this issue of Polar Biology. The studies were conducted in the frame of the National Program of Research in Antarctica (PNRA) of Italy.     

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.     

Nitrogen uptake in the infiltration community, an ice algal community in Antarctic pack-ice

An infiltration community was the dominating ice algal community in pack-ice off Queen Maud Land, Southern Ocean, in January 1993. The community was dominated by autotrophic processes, and the most common species were the prymnesiophyte Phaeocystis antarctica and the diatoms Chaetoceros neglectus and Fragilariopsis cylindrus. The concentration of chlorophyll a was 1.3–47.9 μg l⁻¹, and the inner part of the community was nitrate depleted. Uptake rates of nitrate, nitrite, ammonium, urea and amino acids were measured using ¹⁵N. Nitrate was the major nitrogen source for ice algal growth (67 ± 6% nitrate uptake). It is suggested that % nitrate uptake in the infiltration community decreases during the growth season, from 92% during spring (literature data) to 67% during summer. Scalar irradiance in the infiltration community was high and variable. It reached ca. 2000 μmol m⁻² s⁻¹ at some locations, and nitrate uptake rate was potentially photoinhibited at irradiances >500 μmol m⁻² s⁻¹. Nitrate uptake rate in an average infiltration community (0.6 m of snow cover) was lowered by 13% over a 2-week period due to photoinhibition. Received: 16 December 1996 / Accepted: 5 January 1998     

Affinity for inorganic carbon of Gracilaria tenuistipitata cultured at low and high irradiance

Regulation by irradiance level of the mechanism for dissolved inorganic carbon (DIC) acquisition was examined in the red macroalga Gracilaria tenuistipitata Zhang et Xia. For this purpose, affinity for external DIC, carbonic anhydrase (CA; EC 4.2.1.1) activity and content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) were determined in thalli grown at 45 and 500 μmol photons m⁻² s⁻¹. Oxygen evolution rates declined by 50% when the medium pH was changed from 8.1 to 8.7, and the pH compensation point attained was ca. 9.2. These characteristics were unaffected by the light treatments. In contrast, photosynthetic conductance for DIC at pH 8.7 was doubled in thalli grown at high irradiance compared with those grown at low irradiance (to 0.74 × 10⁻⁶ from 0.33 × 10⁻⁶ m s⁻¹). Photosynthetic rates at saturating DIC concentration were also higher by 60% in thalli grown at high irradiance. These differences could not be attributed to changes in the use of external DIC, since external CA activity did not vary. Although the irradiance level did not modify the pool size of Rubisco, Rubisco content expressed on a chlorophyll a basis was almost doubled at high irradiance. These results likely indicate that the internal transport of DIC towards the active-site of Rubisco, rather than the external use of DIC, is enhanced in the thalli grown at high irradiance. Received: 7 June 1999 / Accepted: 16 October 1999     

Phytoplankton and sea ice algal biomass and physiology during the transition between winter and spring (McMurdo Sound, Antarctica)

The phytoplankton and sea ice algal communities at the end of winter in McMurdo Sound were dominated by Fragilariopsis sublineata, with Thalassiosira antarctica, Melosira adele, Pinnularia quadreata, Entomoneis kjellmannii and heterotrophic dinoflagellates also present. Sea ice algal biomass at the end of winter was very low, only 0.050 ± 0.019 mg chla m⁻² in 2007 and 0.234 ± 0.036 mg chla m⁻² in 2008, but this increased to 0.377 ± 0.078 mg chla m⁻² by early October in 2007 and to 1.07 ± 0.192 by late September in 2008. Under ice phytoplankton biomass remained consistently below 0.1 μg chla l⁻¹ throughout the measuring period in both years. The photosynthetic parameters Fv/Fm, rETRmax and α document microalgal communities that are mostly healthy and well adapted to their low light under ice environment. Our results also suggest that species such as Fragilariopsis sublineata are well adapted to deal with low winter light levels but are unlikely to survive an increase in irradiance, whereas other taxa, such as Thalassiosira antarctica, will do better in a higher light environment.     

Production of Spirulina sp. in sea water supplemented with anaerobic effluents in outdoor raceways under temperate climatic conditions

The use of untreated sea water supplemented with anaerobic effluents from digested pig waste and sodium bicarbonate was evaluated as a low-cost medium for semi-continuous cultivation of a mixed culture of two Spirulina strains in outdoor raceways under temperate climatic conditions (pond temperature in the range 21–26 °C and light intensity in the range 225–957␣μE m⁻² s⁻¹). The mixed culture had a predominant population (86.6 ± 3.9%) of an atypical Spirulina strain consisting of straight filaments, which appeared spontaneously after the strain with helicoidal trichomes had been subcultured. Morphological studies for the identification of the type and size of trichomes of the two strains (HF and SF) were carried out. The proportions of the two strains were observed to be stable during the monitoring period (30 days). Three different sets of semicontinuous cultures were carried out. Sets 1 and 2 were operated under regime 1 (a single addition of anaerobic effluents at time zero and no pH control) during the same season (June and July) of different years. Set 3 was operated under regime 2 (semi-continuous addition of anaerobic effluents and pH control) during the autumn. A minimum productivity of 3.6 g m⁻² day⁻¹ was obtained at one of the lowest temperatures (22.1 °C) and light intensities (245 μE m⁻² s⁻¹) and a maximum productivity of 10.9 g m⁻² day⁻¹ was observed at the highest temperature (25 °C) and highest average light intensity (618 μE m⁻² s⁻¹) registered for sets 1 and 2. The protein content in the Spirulina biomass harvested from these two sets varied from 17% to 65.6%. In set 3, a maximum productivity of 9.0 g m⁻² day⁻¹ was recorded at an average temperature of 24.4 °C and at an average light intensity of 668 μE m⁻² s⁻¹. The protein content in this set under regime 2 varied within a narrower range than in set 1 and set 2 (from 34.8% to 49.1%), apparently because of a continuous availability of ammonia nitrogen at a level of 30–50 mg l⁻¹. However, in terms of the removal of ammonia nitrogen and chemical oxygen demand, regime 1 was more efficient than regime␣2. Received: 3 September 1996 / Received revision: 19 February 1997 / Accepted: 7 March 1997     

Structural and functional properties of sympagic communities in the annual sea ice at Terra Nova Bay (Ross Sea, Antarctica)

Studies on the chemical and biological properties of annual pack ice at a coastal station in Terra Nova Bay (74°41.72′S, 164°11.63′E) were carried out during austral spring at 3-day intervals from 5 November to 1 December 1997. Temporal changes of nutrient concentrations, algal biomasses, taxonomic composition, photosynthetic pigment spectra and P–E relationships were studied. Quantity, composition and degradation rates of organic matter in the intact sea ice were also investigated. In addition, microcosm experiments were carried out to evaluate photosynthetic and photo-acclimation processes of the sympagic flora in relation to different light regimes. High concentrations of ammonia were measured in four ice-cores (weighted mean values of the cores ranged from 4.3 ± 1.9 μM to 7.2 ± 3.4 μM), whereas nitrate and phosphate displayed high concentrations (up to 35.9 μM and 7.6 μM, respectively) only in the bottom layer (135–145 cm depth). Particulate carbohydrate and protein concentrations in the intact sea ice ranged from 0.5 to 2.3 mg l⁻¹ and 0.2 to 2.0 mg l⁻¹, respectively, displaying a notable accumulation of organic matter in the bottom colored layer, where bacterial enzymatic activities also reached the highest values. Aminopeptidase activity was extremely high (up to 19.7 μM l⁻¹ h⁻¹ ± 0.05 in the bottom layer), suggesting a rapid turnover rate of nitrogen–enriched organic compounds (e.g. proteins). By contrast, bacterial secondary production was low, suggesting that only a very small fraction of mobilized organic matter was converted into bacterial biomass (<0.01‰). The sympagic autotrophic biomass (in terms of chlorophaeopigments) of the bottom layer was high, increasing during the sampling period from 680 to 2480 μg l⁻¹. Analyses of pigments performed by HPLC, as well as microscope observations, indicated that diatoms dominated bottom communities. The most important species were Amphiprora sp. and Nitschia cfr. stellata. Bottom sympagic communities showed an average P ^B _max of 0.12 mgC mg Chl⁻¹ and low photoadaptation index (E _k=18 μE m⁻² s⁻¹, E _m=65 μE m⁻² s⁻¹). Results of the microcosm experiment also indicated that communities were photo-oxidized when irradiance exceeded 100 μE m⁻² s⁻¹. This result suggests that micro- autotrophs inhabiting sea ice might have a minor role in the pelagic algal blooms. Accepted: 4 August 1999     

THE INFLUENCE OF FLUCTUATING LIGHT ON DIVERSITY AND SPECIES NUMBER OF NUTRIENT‐LIMITED PHYTOPLANKTON1

The influence of fluctuating light on diversity and species number of a natural phytoplankton assemblage competing for nutrients was investigated for 48 days under semicontinuous culture conditions. Light conditions were either changed periodically from high (65 μmol photons·m^?2·s^?1) to low intensity (15 μmol photons·m^?2·s^?1) at intervals of 1, 3, 6, and 12 days or fixed at constant light conditions of intermediate intensity (40 μmol photons·m^?2·s^?1). Fluctuating light at intervals of 1–12 days significantly affected phytoplankton diversity. The development of phytoplankton communities differed in treatments with different light regimes. In treatments with long light intervals, species abundance oscillated with the light phases. Differences in the temporal development of phytoplankton communities resulted in hump‐shaped relations between the interval length of the light phases and both species number and diversity index and can be explained by the intermediate disturbance hypothesis. Fluctuating light tends to sustain phytoplankton diversity under nutrient limitation if the light regime changes in the order of several days. This indicates that temporal changes in weather regime are important in preventing competitive exclusion of phytoplankton species in nature.     

Unusually low carbon isotope ratios in plants from hanging gardens in southern Utah

Leaf carbon isotope ratios (δ¹³C) and photosynthetic gas exchange were measured on plants growing in hanging garden communities in southern Utah, USA. Hanging gardens are unusual, mesic cliff communities occurring where water seeps from the sandstone bedrock in an otherwise extremely arid region; there is very limited overlap in species distributions inside and outside these gardens. Solar exposure in hanging gardens varied with orientation and one of the gardens (Ribbon Garden) was shaded throughout the day. The leaf δ¹³C values of plants in hanging gardens were significantly more negative than for plants from either nearby ephemeral wash or riparian communities. In Ribbon Garden, the observed δ¹³C values were as low as −34.8‰, placing them among the most negative values reported for any terrestrial plant species growing in a natural environment. Hanging garden plants were exposed to normal atmospheric CO₂ with an average δ¹³C value of −7.9‰ and so the low leaf δ¹³C values could not be attributed to exposure to a CO₂ source with low ¹³C content. There was a seasonal change toward more negative leaf δ¹³C values at the end of the growing season. The observed leaf δ¹³C values were consistent with photosynthetic gas exchange measurements that indicated unusually high leaf intercellular CO₂ concentrations associated with the relatively low light levels in hanging gardens. Thus, extremely negative leaf δ¹³C values would be expected if significant amounts of the seasonal carbon gain occur at light levels low enough to be near the light compensation point. Maximum observed photosynthetic rates varied with light levels at each of the gardens, with maximum rates averaging 20.3, 14.6, and 3.1 μmol m⁻²s⁻¹ at Double Garden, Lost Garden, and Ribbon Garden, respectively. Leaf nitrogen contents averaged 18.5 mg g⁻¹ in species from the more shaded hanging gardens (Lost and Ribbon). When expressed on a leaf area basis, nitrogen contents averaged 117 mmol N m⁻² at Lost Garden and 65 mmol N m⁻² at Ribbon Garden (shadiest of the two gardens). Leaf nitrogen isotope ratios averaged −2.3‰ (range of −0.7 to −6.1‰), suggesting that most of the nitrogen was derived from a biological fixation source which is most likely the Nostoc growing on the sandstone walls at the seep. These values contrast with leaf nitrogen isotope ratios of 5–9‰ which have been previously reported for arid zone plants in nearby ecosystems. Received: 19 January 1997 / Accepted: 19 April 1997