IDENTIFICATION OF CYLINDROSPERMOPSIN IN APHANIZOMENON OVALISPORUM (CYANOPHYCEAE) ISOLATED FROM LAKE KINNERET,ISRAEL1

Aphanizomenon ovalisporum (Forti) was identified and isolated from Lake Kinneret upon its first appearance as a dominant bloom in late 1994. This cyanobacterial species, not previously known to be toxic, was evaluated by a commonly used mouse bioassay and was demonstrated to induce toxic symptoms that were distinguishable from the typical symptoms of the neurotoxins previously reported in Aphanizomenon flos-aquae (L.) Ralfs. Mice died 5–24 h after crude extracts were injected intraperitoneally, and the LD₅₀ value was estimated as 465 mg dry wt biomass · kg^?1 mouse. A toxicity-guided fractionation of the active extract indicated that the potent substance is polar an nature. The structure of the active compound was determined by its mass spectrometry and NMR data. The compound was found to be the sulfate-guanidinium zwitterion, cylindrospermopsin, previously isolated from the cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) and recently also reported in Umezakia natans (Watanabe). This is the first time that Aphanizomenon ovalisporum has been reported to contain a toxic compound.     

DEVELOPMENT AND GERMINATION OF AKINETES OF APHANIZOMENON FLOS-AQUAE1

Flakes of Aphanizomenon flos-aquae collected from an ice-covered lake were found to contain all developmental stages from vegetative cells to mature akinetes. Changes during development include increase in cell size, gradual disappearance of gas vacuoles (clusters of gas vesicles), narrowing of intrathylakoidal spaces, and increase in cytoplasmic density. Development of akinetes is accompanied by proliferation of ribosomes, including polyribosomes, cyanophycin granules (structured, granules), and glycogen granules. The lipid bodies of vegetative cells are reduced in size and number in mature akinetes. Akinetes may occur singly or as multiples in sequence in a filament, either terminal or intercalary. Loss of flotation by increase in cytoplasmic density permits filaments to sink and overwinter in bottom sediments. The sequence was found to be reversed during germination of akinetes. Cyanophycin granules are reduced in size and staining density in the sporelings, and very few glycogen granules are seen. Gas vesicles reappear and increase in number, and intrathylakoidal spaces become wider. These changes then would permit the sporelings to rise from the bottom and begin another season's bloom.     

MOLECULAR AND PHYLOGENETIC CHARACTERIZATION OF PHORMIDIUM SPECIES (CYANOPROKARYOTA) USING THE CPCB‐IGS‐CPCA LOCUS1

The accurate determination of species of Cyanoprokaryota/Cyanophyceae has many important applications. These include the assessment of risk with regard to blooms in water reservoirs as well as the identification of species capable of producing valuable bioactive compounds. Commonly, Cyanoprokaryota are classified based on their morphology. However, morphological criteria are not always reliable because they may change, for example, due to environmental factors. Thus, genetic and molecular analyses are a promising additional approach, but their application has so far been limited to relatively few genera. In light of this, we present here the first characterization of species and strains of the genus Phormidium Kütz. based on the cpcB‐IGS‐cpcA locus of the phycocyanin operon. In phylogenetic analyses using deduced amino acid sequences of the cpcB‐cpcA regions, Phormidium was found to be polyphyletic. This analysis appeared to be dominated by the cpcB region, which is characterized by a relatively high percentage of informative substitutions. The percentage of variable positions within the cpcB‐IGS‐cpcA locus overall was 16.5%, thereby indicating a level of divergence remarkably higher than that reported for Nodularia and Arthrospira in previous studies relying on cpcB‐IGS‐cpcA. Further, alignment of informative nucleotide substitutions in the cpcB‐IGS‐cpcA sequences revealed a mosaic distribution, which may be indicative of genetic recombination events. Finally, the length and sequences of the IGS region alone proved useful as markers to differentiate the cyanobacterial genus Phormidium. However, whether the IGS region per se is sufficiently discriminatory to differentiate between Phormidium species or even strains requires further investigation using newly identified Phormidium sequence data.     

EFFECT OF TEMPERATURE ON GERMINATION OF RHIZOCLONIUM RIPARIUM (SIPHONOCLADALES,CHLOROPHYTA) AKINETES AND ZOOSPORES1

The temperature requirements for germination by reproductive initials of Rhizoclonium riparium (Roth) Harv., a filamentous green alga, were investigated in laboratory culture. Akinetes and zoospores were produced by exposing aging cultures to high temperatures (40°C). Germination proceeded rapidly and followed a typical bell-shaped response curve, with germination optima between 15 and 20°C. These findings follow the trend found in related algae, i.e. reproductive initials are produced under stressful conditions.     

STRESS‐INDUCED FILAMENT FRAGMENTATION OF CALOTHRIX ELENKINII (CYANOBACTERIA) IS FACILITATED BY DEATH OF HIGH‐FLUORESCENCE CELLS1

Irradiance power and spectral composition as well as nutrient availability strongly influence differentiation of filamentous cyanobacteria. When monitoring the life cycle of Calothrix elenkinii Kossinsk., we found that low nitrogen concentration and growth under green light led to a transient appearance of high‐fluorescence cells that rapidly bleach and disintegrate, thus breaking the parental filament into shorter parts. The dynamics of the process were monitored in a microscope growth chamber by measuring transmission and chl fluorescence of individual cells by a high‐sensitivity camera. Typically, the high‐fluorescence cells appeared near the center of the parental trichome signaled by a rapid 2‐ to 3‐fold rise in their fluorescence emission. By measuring the fluorescence excitation spectra with resolution of individual cells, we showed that the elevated fluorescence emission was largely due to a high absorption by phycoerythrin and energy transfer to chl. Typically, after no more than 20 min, the fluorescence abruptly disappeared with transmission images, indicating loss of pigmentation. The bleaching was a natural process that was not caused by the measuring light. Depending on the mechanical strain, the cell bleaching was followed by breaking of the parental filament. We propose that the high‐fluorescence cells appear as a phase of programmed cell death, allowing the fragmented filaments to escape from unfavorable environmental conditions.     

THE IMPORTANCE OF SHALLOW SEDIMENTS IN THE RECRUITMENT OF ANABAENA AND APHANIZOMENON (CYANOPHYCEAE)1

Recruitment of Anabaena and Aphanizomenon from the sediments to the water column was investigated in shallow (1–2 m) and deep (6–7 m) areas of Lake Limmaren, central Sweden. Recruitment traps attached to the bottom were sampled weekly throughout the summer season (June through September). A comparison between the two sites shows that the largest part of the recruited cells originated from the shallow site, although recruitment occurred at all depths in the lake. There were also differences between the species, regarding the site as well as the timing of the recruitment. The contribution of the inoculum to the pelagic population was calculated to vary between 0.003% and 0.05% for the different species. From these results we conclude that shallow sediments are more important than deep ones for the recruitment and that the inoculum in Lake Limmaren is small but may still be an important factor in the population dynamics.     

OXYGEN-INDUCED CHANGES IN MORPHOLOGY OF AGGREGATES OF APHANIZOMENON FLOS-AQUAE (CYANOPHYCEAE): IMPLICATIONS FOR NITROGEN FIXATION POTENTIALS1

The filamentous nitrogen-fixing cyanobacterium Aphanizomenon flos-aquae (L.) Ralfs forms bundle or fake shaped aggregates which can provide buoyancy control, protection against intense illumination, enhancement of phycosphere nutrient regeneration, and which may result from size-selective herbivory by zooplankton. The dimensions of aggregates can change quickly. In this study, after a period of darkness, illumination caused aggregates to elongate approximately five-fold over a 10-15 min period. The metamorphosis was reversible upon cessation of illumination and through successive light-dark cycles. Manipulations of environmental oxygen concentration and photosystem Ü activity (via DCMU amendment), together with measurements made inside flakes with O₂-sensitive microelectrocles, showed that the metamorphosis was a response to oxygen concentration and operated to enhance diffusive efflux of photosynthetically produced oxygen during illumination. During darkness oxygen concentration within contracted aggregates became severely depleted relative to the environment. We propose that metamorphic minimization of local oxygen concentration is an adaptation that enhances the ability of Aphanizomenon flos-aquae to fix atmospheric nitrogen via the oxygen-labile nitrogenase enzyme system.     

ALGAL PHOTOSYNTHETIC MEMBRANE COMPLEXES AND THE PHOTOSYNTHESIS-IRRADIANCE CURVE: A COMPARISON OF LIGHT-ADAPTATION RESPONSES IN CHLAMYDOMONAS REINHARDTII (CHLOROPHYTA)1

Chlamydomonas reinhardtii was grown at photon flux densities (PFDs) ranging from 47 to 400 μE.m^-2 s^-1. The total cellular content of chlorophyll (Chl) was twice as high in the low light (LL) versus high light (HL) grown cells. On an equal Chl basis, photosystem II (PSII) and cytochrome f (Cyt f) content was higher in HL cells, but photosystem I (PSI) concentration displayed little variation with the light intensity during cell growth. Consequently, there was a shift in the ratio of PSII / PSI and Cyt / PSI from near unity in LL cells to greater than two in HL cells. The functional Chl antenna size of PSII and PSI ranged from 460 and 170 Chl (a + b)in HL-grown cells to 620 and 370 Chl (a+ b)in LL-grown cells, respectively. The initial slope of the Chl-specific photosyn-thesis-irradiance (P-I) curve was similar in LL- and HL-grown cells, but the light saturated rate of photosynthesis was lower under LL. The response to low light was beneficial at the cellular level, since there was an enhancement of photosynthesis in LL. The PFD for the onset of light saturation, 1 was a factor of 2 lower in LL- relative to HL-grown photosythetic membranes. Since growth PFD varied by a factor of ten, photosynthesis shifted from being light-limited in the LL regime to light-saturated in the HL regime. The requirement for balanced absorption of light by the two photosystems constrains the PSII / PSI ratio to near unity when growth is light-limited, but such a constraint does not apply in HL conditions. Instead the concentration of individual electron transport complexes way be related to the pool size necessary for maximum rates of steady-state electron transport. Thus the stoichiometry of electron transport complexes changes in response to growth PFD and this change is correlated with the response flexlbility of algal photosynthesis in diverse light environments.     

POLYPHASIC CHARACTERIZATION OF THREE STRAINS OF ANABAENA RENIFORMIS AND APHANIZOMENON APHANIZOMENOIDES (CYANOBACTERIA) AND THEIR RECLASSIFICATION TO SPHAEROSPERMUM GEN. NOV. (INCL. ANABAENA KISSELEVIANA)1

Occurrences of rare cyanobacteria Anabaena reniformis Lemmerm. and Aphanizomenon aphanizomenoides (Forti) Horecká et Komárek were recently detected at several localities in the Czech Republic. Two monoclonal strains of An. reniformis and one strain of Aph. aphanizomenoides were isolated from distant localities and different sampling years. They were characterized by a combination of morphological, genetic, and biochemical approaches. For the first time, partial 16S rRNA gene sequences were obtained for these morphospecies. Based on this gene, all of these strains clustered separately from other planktonic Anabaena and Aphanizomenon strains. They appeared in a cluster with Cylindrospermopsis Seenaya et Subba Raju and Raphidiopsis F. E. Fritsch et M. F. Rich, clustered closely together with two An. kisseleviana Elenkin strains available from GenBank. A new generic entity was defined (Sphaerospermum gen. nov., with the type species S. reniforme, based on the traditional species An. reniformis). These results contribute significantly to the knowledge base about genetic heterogeneity among planktonic Anabaena–like and Aphanizomenon–like morphospecies. Accordingly, the subgenus Dolichospermum, previously proposed for the group of planktonic Anabaena, should be revaluated. Secondary metabolite profiles of the An. reniformis and Aph. aphanizomenoides strains differed considerably from 17 other planktonic Anabaena strains of eight morphospecies isolated from Czech water bodies. Production of puwainaphycin A was found in both of the An. reniformis strains. Despite the relatively short phylogenetic distance from Cylidrospermopsis, the production of cylindrospermopsin was not detected in any of our strains.     

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

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

PHOTOSYNTHETIC LIGHT-HARVESTING FUNCTION OF VIOLAXANTHIN IN NANNOCHLOROPSIS SPP. (EUSTIGMATOPHYCEAE)1

Whole cell absorption spectra of the Eustigmatophycean algae Nannochloropsis salina Bourrelly and Nannochloropsis sp. reveal the presence of a distinct absorption peak at 490 nm. The lack of chlorophylls b and c in these species indicates that this peak must be attributed to carotenoid absorption. In vivo fluorescence excitation spectra for chlorophyll a emission show a corresponding maximum at 490 nm. This peak is more clearly resolved than carotenoid maxima in other algal classes due to the absence of accessory chlorophylls. The carotenoid composition of the two Nannochloropsis species shows that violaxanthin and vaucheriaxanthin are the main contributors to 490 nm absorption. Violaxanthin accounts for approximately 60% of the total carotenoid in both clones. We conclude that light absorption by violaxanthin, and possibly by vaucheriaxanthin, is coupled in energy transfer to chlorophyll a and that violaxanthin is the major light-harvesting pigment in the Eustigmatophyceae. This is the first report of the photosynthetic light-harvesting function of this carotenoid.     

PHOTOSYNTHETIC LIGHT-HARVESTING FUNCTION OF VIOLAXANTHIN IN NANNOCHLOROPSIS SPP. (EUSTIGMATOPHYCEAE)1

Whole cell absorption spectra of the Eustigmatophycean algae Nannochloropsis salina Bourrelly and Nannochloropsis sp. reveal the presence of a distinct absorption peak at 490 nm. The lack of chlorophylls b and c in these species indicates that this peak must be attributed to carotenoid absorption. In vivo fluorescence excitation spectra for chlorophyll a emission show a corresponding maximum at 490 nm. This peak is more clearly resolved than carotenoid maxima in other algal classes due to the absence of accessory chlorophylls. The carotenoid composition of the two Nannochloropsis species shows that violaxanthin and vaucheriaxanthin are the main contributors to 490 nm absorption. Violaxanthin accounts for approximately 60% of the total carotenoid in both clones. We conclude that light absorption by violaxanthin, and possibly by vaucheriaxanthin, is coupled in energy transfer to chlorophyll a and that violaxanthin is the major light-harvesting pigment in the Eustigmatophyceae. This is the first report of the photosynthetic light-harvesting function of this carotenoid.     

CHARACTERIZATION OF PSYCHROPHILIC OSCILLATORIANS (CYANOBACTERIA) FROM ANTARCTIC MELTWATER PONDS

Oscillatorian cyanobacteria dominate benthic microbial mat communities in many polar freshwater ecosystems. Capable of growth at low temperatures, all benthic polar oscillatorians characterized to date are psychrotolerant (growth optima > 15° C) as opposed to psychrophilic (growth optima ≤ 15° C). Here, psychrophilic oscillatorians isolated from meltwater ponds on Antarctica's McMurdo Ice Shelf are described. Growth and photosynthetic rates were investigated at multiple temperatures, and compared with those of a psychrotolerant isolate from the same region. Two isolates showed a growth maximum at 8° C, with rates of 0.12 and 0.08 doublings·d ^{? 1}, respectively. Neither displayed detectable growth at 24° C. The psychrotolerant isolate showed almost imperceptible growth at 4° C and a rate of 0.9 doublings·d ^{? 1} at its optimal temperature of ～23° C. In both photosynthesis versus irradiance and photosynthesis versus temperature experiments, exponentially growing cultures were acclimated for 14 days at 3, 8, 12, 20, and 24° C under saturating light intensity, and [¹⁴C] photoincorporation rates were measured. Psychrophilic isolates acclimated at 8° C showed greatest photosynthetic rates; those acclimated at 3° C were capable of active photosynthesis, but photoincorporation was not detected in cells acclimated at 20 and 24° C, because these isolates were not viable after 14 days at those temperatures. The psychrotolerant isolate, conversely, displayed maximum photosynthetic rates at 24° C, though photoincorporation was actively occurring at 3° C. Within acclimation temperature treatments, short‐term photosynthetic rates increased with increasing incubation temperature for both psychrophilic and psychrotolerant isolates. These results indicate the importance of temperature acclimation before assays when determining optimal physiological temperatures. All isolates displayed photosynthetic saturation at low light levels (<128 μmol·m ^{? 2}·s ^{? 1}) but were not photoinhibited at the highest light treatment (233 μmol·m ^{? 2}·s ^{? 1}). Field studies examining the impact of temperature on photosynthetic responses of intact benthic mats, under natural solar irradiance, showed the mat communities to be actively photosynthesizing from 2 to 20° C, with maximum photoincorporation at 20° C, as well as capable of a rapid response to an increase in temperature. The rarity of psychrophilic cyanobacteria, relative to psychrotolerant strains, may be due to their extremely slow growth rates and inability to take advantage of occasional excursions to higher temperatures. We suggest an evolutionary scenario in which psychrophilic strains, or their most recent common ancestor, lost the ability to grow at higher temperatures while maintaining a broad tolerance for fluctuations in other physical and chemical parameters that define shallow meltwater Antarctic ecosystems.     

RELATIONSHIP BETWEEN PHOTOSYNTHETIC OXYGEN CYCLING AND CARBON ASSIMILATION IN SYNECHOCOCCUS WH7803 (CYANOPHYTA)1

Mass spectrometric analysis of oxygen uptake and evolution in the light by marine Synechococcus WH7803 indicated that the respiration rate was near zero at low irradiance levels but increased significantly at high irradiances. The light intensity (I_r) at which oxygen uptake began to increase with increasing light intensity depended on the growth irradiance of the culture. In each case, I_r coincided with the minimum light intensity for saturation of carbon assimilation (I_k). At irradiances >I_r, net oxygen evolution rates paralleled carbon assimilation rates. Oxygen uptake at high light intensities was inhibited by DCMU, indicating that oxygen uptake was due to Mehler reaction activity. The onset of Mehler activity at I_k supports the idea that oxygen becomes an alternative sink for electrons from photosystem I when NADPH turnover is limited by the capacity of the dark reactions to utilize reductant.     

PHOTOSYNTHETIC RESPONSE OF THE GIANT KELP MACROCYSTIS PYRIFERA (PHAEOPHYCEAE) TO ULTRAVIOLET RADIATION1

Solar ultraviolet radiation (UVA + UVB) impairs photosynthesis in marine algae. Canopy blades of the giant kelp Macrocystis pyrifera (L.) C. Agardh are exposed to high levels of solar UV in the field. To determine the effects of UV radiation on photosynthesis in the giant kelp and to identify sites of UV damage, O₂ evolution, reaction center organization, light harvesting, and energy transfer efficiency were measured in canopy blades that had been exposed to elevated levels of UV in the laboratory. UV treatment reduced both the light-saturated rate and the light-limited rate of photosynthesis by 50% but produced no significant change in the rate of dark respiration. A significant impairment of photosystem II (PSII) reaction center function was observed, suggesting that PSII is a major site of damage in chromophytes. Reduced quantum efficiency of photosynthesis and loss of energy transfer from light-harvesting pigments (fucoxanthin, chlorophyll a, and chlorophyll c) to PSII indicate that the major light-harvesting complex of M. pyrifera, the fucoxanthin-chlorophyll protein complex (FCPC), was another site of UV damage. These measures provide the first evidence of a direct effect of UV radiation on specific sites in the photosynthetic apparatus of chromophytes and indicate that in situ fluorescence excitation analysis may be a simple means to detect UV stress in algae.     

AUTOINHIBITION OF SPORE GERMINATION IN NOSTOC SPONGIAEFORME (CYANOPHYCEAE)1

Medium in which cultures of Nostoc spongiaeforme Ag. have sporulated contains one or more substances which inhbit the germination of spores (akinetes) of this organism. Germination of spores occurs rapidly in the absence of these substances, but is virtually completely suppressed in their presence.     

IMPACT OF IRON LIMITATION ON THE PHOTOSYNTHETIC APPARATUS OF THE DIATOM CHAETOCEROS MUELLERI (BACILLARIOPHYCEAE)

Iron starvation induced marked increases in flavodoxin abundance and decreases in light-saturated and light-limited photosynthesis rates in the diatom Chaetoceros muelleri. Consistent with the substitution of flavodoxin for ferredoxin as an early response to iron starvation, increases of flavodoxin abundance were observed before declines of cell division rate or chl a specific photosynthesis rates. Changes in the abundance of flavodoxin after the addition of iron to iron-starved cells indicated that flavodoxin was not actively degraded under iron-replete conditions. Greater declines in light-saturated oxygen evolution rates than dark oxygen consumption rates indicated that the mitochondrial electron transfer chain was not affected as greatly by iron starvation as the photosynthetic electron transfer chain. The carbon:nitrogen ratio was unaffected by iron starvation, suggesting that photosynthetic electron transfer was a primary target of iron starvation and that reductions in nitrate assimilation were due to energy limitation (the C:N ratio would be expected to rise under nitrogen-limited but energy-replete conditions). Parallel changes were observed in the maximum light-saturated photosynthesis rate and the light-limited initial slope of the photosynthesis-light curve during iron starvation and recovery. The lowest photosynthesis rates were observed in iron-starved cells and the highest values in iron-replete cells. The light saturation parameter, I_k, was not affected by iron starvation, nor was the chl-to-C ratio markedly reduced. These observations were consistent with iron starvation having a similar or greater effect on photochemical charge separation in PSII than on downstream electron transfer steps. Declines of the ratio of variable to maximum fluorescence in iron-starved cells were consistent with PSII being a primary target of iron starvation. The functional cross-section of PSII was affected only marginally (<20%) by iron starvation, with the largest values observed in iron-starved cells. The rate constant for electron transfer calculated from fast repetition rate fluorescence was found to covary with the light-saturated photosynthesis rate; it was lowest in the most severely starved cells.     

FLUORESCENCE INDUCTION AND PHOTOSYNTHETIC RESPONSES OF ARCTIC ICE ALGAE TO SAMPLE TREATMENT AND SALINITY1

The measurement of Photosynthetic rates of algae growing on the undersurface of 1. 7 m thick ice in the Canadian Arctic (Resolute Passage. N.W.T.) presents several problems. During the preparation of samples for physiological measurements, the ice algae may he exposed to salinity and temperature shocks. Fluorescence induction (the rise in in vivo Chl a fluorescence intensity during a period of millineconds) and photosynthesis-irradiance (PI) experiments examined the potential effects of salinity and temperature on the physiology of ice algae. Experimental suspensions were routinely prepared by scraping one part ire crystals (11–14%₀ salinity) and attached algae from the bottom ice into four parts filtered seawater (32%₀ salinity). giving a final salinity of 28–31%₀. Post-dilution of melted ice scrapings with seawater suppressed photosynthetic ¹⁴C-fixation and decreased A_DCMU (the area above the fluorescence induction curve measured in the presence of the inhibitor DCMC: an estimate of photosynthetic capacity) by a factor of 3–16. due to the low salinity of the melted ice scrapings. Fluorescence induction and PI experiments showed that the ice algae had a salinity optimum near 30%₀, close to the ambient seawater salinity, Experiments in which the Chl a concentration was manipulated showed that A_DCMU, P^a_m (Chl a-normalized rate of photosynthesis at light saturation), and a (photosynthetic efficiency) declined with increasing Chl a concentration. Ice algae tolerated heating (l.5°C-min^-1) up to 17° C, above which A_DCMU’decreased with sample temperature.     

EFFECT OF NITRATE CONCENTRATION ON THE RELATIONSHIP BETWEEN PHOTOSYNTHETIC OXYGEN EVOLUTION AND ELECTRON TRANSPORT RATE IN ULVA RIGIDA (CHLOROPHYTA)1

The electron transport rate (ETR) versus gross photosynthesis (GPS) relationship varies as a function of species, temperature, irradiance, and inorganic carbon levels, but less is known about the effect of nitrogen supply on this relationship. The objective of this study was to evaluate the effect of nitrate concentration on the ETR versus GPS relationship in Ulva rigida C. Agardh from the Mediterranean Sea. Chlorophyll content and tissue absorptance increased 2‐fold as nitrate in the media increased from 0 to 50 μM. Whereas internal N content increases 3‐fold at 50 μM, internal C increased slightly. Oxygen evolution and ETR, evaluated as in vivo chl fluorescence using pulse amplitude modulated fluorometry, in general saturated at irradiances above 100 μmol photons·m^?2·s^?1. Both maximum ETR and GPS values increased as nitrate concentration increased. In general, the ETR versus GPS relationship showed a linear response to increasing nitrate with little variance of the data. This relationship, however, became more variable at high irradiances and high nitrate concentrations. The ETR/GPS ratio was close to the theoretical value of 4 at low nitrate concentrations, and the ratio decreased exponentially when nitrate concentration in the media increased. The variations of ETR/GPS under different inorganic nitrogen supply are discussed in terms of the effect of nitrate on the photosynthesis and respiration relationship.     

LIGHT DEPENDENCY OF PHOTOSYNTHETIC RECOVERY DURING WETTING AND THE ACCLIMATION OF PHOTOSYNTHETIC APPARATUS TO LIGHT FLUCTUATION IN A TERRESTRIAL CYANOBACTERIUM NOSTOC COMMUNE1

The PSII photochemical activity in a terrestrial cyanobacterium Nostoc commune Vaucher ex Bornet et Flahault during rewetting was undetectable in the dark but was immediately recognized in the light. The maximum quantum yield of PSII (F_v/F_m) during rewetting in the light rose to 85% of the maximum within ～30 min and slowly reached the maximum within 6 h, while with rewetting in the darkness for 6 h and then exposure to light the recovery of F_v/F_m required only ～3 min. These results suggested that recovery of photochemical activity might depend on two processes, light dependence and light independence, and the activation of photosynthetic recovery in the initial phase was severely light dependent. The inhibitor experiments showed that the recovery of F_v/F_m was not affected by chloramphenicol (CMP), but severely inhibited by 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea (DCMU) in the light, suggesting that the light‐dependent recovery of photochemical activity did not require de novo protein synthesis but required activation of PSII associated with electron flow to plastoquinone. Furthermore, the test indicated that the lower light intensity and the red light were of benefit to its activation of photochemical activity. In an outdoor experiment of diurnal changes of photochemical activity, our results showed that PSII photochemical activity was sensitive to light fluctuation, and the nonphotochemical quenching (NPQ) was rapidly enhanced at noon. Furthermore, the test suggested that the repair of PSII by de novo protein synthesis played an important role in the acclimation of photosynthetic apparatus to high light, and the heavily cloudy day was more beneficial for maintaining high photochemical activity.