NITROGEN FIXATION,HYDROGEN CYCLING,AND ELECTRON TRANSPORT KINETICS IN TRICHODESMIUM ERYTHRAEUM (CYANOBACTERIA) STRAIN IMS1011

This study describes the relationships between dinitrogen (N₂) fixation, dihydrogen (H₂) production, and electron transport associated with photosynthesis and respiration in the marine cyanobacterium Trichodesmium erythraeum Ehrenb. strain IMS101. The ratio of H₂ produced:N₂ fixed (H₂:N₂) was controlled by the light intensity and by the light spectral composition and was affected by the growth irradiance level. For Trichodesmium cells grown at 50 μmol photons · m^?2 · s^?1, the rate of N₂ fixation, as measured by acetylene reduction, saturated at light intensities of 200 μmol photons · m^?2 · s^?1. In contrast, net H₂ production continued to increase with light levels up to 1,000 μmol photons · m^?2 · s^?1. The H₂:N₂ ratios increased monotonically with irradiance, and the variable fluorescence measured using a fast repetition rate fluorometer (FRRF) revealed that this increase was accompanied by a progressive reduction of the plastoquinone (PQ) pool. Additions of 2,5‐dibromo‐3‐methyl‐6‐isopropyl‐p‐benzoquinone (DBMIB), an inhibitor of electron transport from PQ pool to PSI, diminished both N₂ fixation and net H₂ production, while the H₂:N₂ ratio increased with increasing level of PQ pool reduction. In the presence of 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea (DCMU), nitrogenase activity declined but could be prolonged by increasing the light intensity and by removing the oxygen supply. These results on the coupling of N₂ fixation and H₂ cycling in Trichodesmium indicate how light intensity and light spectral quality of the open ocean can influence the H₂:N₂ ratio and modulate net H₂ production.     

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.     

Dynamic responses of photosystem II and phycobilisomes to changing light in the cyanobacterium Synechococcus sp. PCC 7942

We have examined the molecular and photosynthetic responses of a planktonic cyanobacterium to shifts in light intensity over periods up to one generation (7 h). Synechococcus sp. PCC 7942 possesses two functionally distinct forms of the D1 protein, D1∶1 and D1∶2. Photosystem II (PSII) centers containing D1∶1 are less efficient and more susceptible to photoinhibition than are centers containing D 1∶2. Under 50 μmol photons· m^?2·s^?1, PSII centers contain D1∶1, but upon shifts to higher light (200 to 1000 μmol photons·m^?2·s^?1), D1∶1 is rapidly replaced by D 1∶2, with the rate of interchange dependent on the magnitude of the light shift. This interchange is readily reversed when cells are returned to 50 μmol photons·m^?2·s^?1. If, however, incubation under 200 μmol photons·m^?2·s^?1 is extended, D1∶1 content recovers and by 3 h after the light shift D1∶1 once again predominates. Oxygen evolution and chlorophyll (Chl) fluorescence measurements spanning the light shift and D1 interchanges showed an initial inhibition of photosynthesis at 200 μmol photons·m^?2·s^?1, which correlates with a proportional loss of total D1 protein and a cessation of growth. This was followed by recovery in photosynthesis and growth as the maximum level of D 1∶2 is reached after 2 h at 200 μmol photons·m^?2·s^?1. Thereafter, photosynthesis steadily declines with the loss of D1∶2 and the return of the less-efficient D1∶1. During the D1∶1/D1∶2 interchanges, no significant change occurs in the level of phycocyanin (PC) and Chl a, nor of the phycobilisome rod linkers. Nevertheless, the initial PC/Chl a ratio strongly influences the magnitude of photo inhibition and recovery during the light shifts. In Synechococcus sp. PCC 7942, the PC/Chl a ratio responds only slowly to light intensity or quality, while the rapid but transient interchange between D1∶1 and D 1∶2 modulates PSII activity to limit damage upon exposure to excess light.     

CYANOBACTERIAL ACCLIMATION TO RAPIDLY FLUCTUATING LIGHT IS CONSTRAINED BY INORGANIC CARBON STATUS1

Acclimation to rapidly fluctuating light, simulating shallow aquatic habitats, is altered depending on inorganic carbon (C_i) availability. Under steady light of 50 μmol photons·m^?2·s^?1, the growth rate of Synechococcus elongatus PCC7942 was similar in cells grown in high C_i (4 mM) and low C_i (0.02 mM), with induced carbon concentrating mechanisms compensating for low C_i. Growth under fluctuating light of a 1‐s period averaging 50 μmol photons·m^?2·s^?1 caused a drop in growth rate of 28%±6% in high C_i cells and 38%±8% in low C_i cells. In high C_i cells under fluctuating light, the PSI/PSII ratio increased, the PSII absorption cross‐section decreased, and the PSII turnover rate increased in a pattern similar to high‐light acclimation. In low C_i cells under fluctuating light, the PSI/PSII ratio decreased, the PSII absorption cross‐section decreased, and the PSII turnover remained slow. Electron transport rate was similar in high and low C_i cells but in both was lower under fluctuating than under steady light. After acclimation to a 1‐s period fluctuating light, electron transport rate decreased under steady or long‐period fluctuating light. We hypothesize that high C_i cells acclimated to exploit the bright phases of the fluctuating light, whereas low C_i cells enlarged their PSII pool to integrate the fluctuating light and dampen the variation of the electron flux into a rate‐restricted C_i pool. Light response curves measured under steady light, widely used to predict photosynthetic rates, do not properly predict photosynthetic rates achieved under fluctuating light, and exploitation of fluctuating light is altered by C_i status.     

PHOTOSYNTHESIS IN AN EXTREME SHADE ENVIRONMENT: BENTHIC MICROBIAL MATS FROM LAKE HOARE, A PERMANENTLY ICE-COVERED ANTARCTIC LAKE

We investigated the composition of benthic microbial mats in permanently ice-covered Lake Hoare, Antarctica, and their irradiance vs. photosynthetic oxygen exchange relationships. Mats could be subdivided into three distinct depth zones: a seasonally ice-free “moat” zone and two under-ice zones. The upper under-ice zone extended from below the 3.5 m thick ice to approximately 13 m and the lower from below 13 m to 22 m. Moat mats were acclimated to the high irradiance they experienced during summer. They contained photoprotective pigments, predominantly those characteristic of cyanobacteria, and had high compensation and saturating irradiances (E_c and E_k) of 75 and 130 μmol photons·m⁻²·s⁻¹, respectively. The moat mats used light inefficiently. The upper under-ice community contained both cyanobacteria and diatoms. Within this zone, biomass (as pigments) increased with increasing depth, reaching a maximum at 10 m. Phycoerythrin was abundant in this zone, with shade acclimation and efficiency of utilization of incident light increasing with depth to a maximum of 0.06 mol C fixed·mol⁻¹ incident photons under light-limiting conditions. Precipitation of inorganic carbon as calcite was associated with this community, representing up to 50% of the carbon sequestered into the sediment. The lower under-ice zone was characterized by a decline in pigment concentrations with depth and an increasing prevalence of diatoms. Photosynthesis in this community was highly shade acclimated and efficient, with E_c and E_k below 0.5 μmol·m⁻²·s⁻¹ and 2 μmol·m⁻²·s⁻¹, respectively, and maximum yields of 0.04 mol C fixed·mol⁻¹ incident quanta. Carbon uptake in situ by both under-ice and moat mats was estimated at up to 100 and 140 mg·m⁻²·day⁻¹, based on the photosynthesis–irradiance curves, incident irradiance, and light attenuation by ice and the water column.     

PHOTOINHIBITION OF MECHANICALLY STIMULABLE BIOLUMINESCENCE IN THE AUTOTROPHIC DINOFLAGELLATE CERATIUM FUSUS (PYRROPHYTA)1

Ceratium fusus (Ehrenb.) Dujardin was exposed to light of different wavelengths and photon flux densities (PFDs) to examine their effects on mechanically stimulable bioluminescence (MSL). Photoinhibition of MSL was proportional to the logarithm of PFD. Exposure to I μmol photons·m^?2s^?1 of broadband blue light (ca. 400–500 nm) produced near-complete photoinhibition (≥90% reduction in MSL) with a threshold at ca. 0.01 μmol photons·m^?2·s^?1. The threshold of photoinhibition was ca. an order of magnitude greater for both broadband green (ca. 500–580 nm) and red light (ca. 660–700 nm). Exposure to narrow spectral bands (ca. 10 nm half bandwidth) from 400 and 700 nm at a PFD of 0.1 μmol photons·m^?2·s^?1 produced a maximal response of photoinhibition in the blue wavelengths (peak ca. 490 nm). A photoinhibition response (≥ 10%) in the green (ca. 500–540 nm) and red wavelengths (ca. 680 nm) occurred only at higher PFDs (1 and 10 μmol photons·m^?2·s^?1). The spectral response is similar to that reported for Gonyaulax polyedra Stein and Pyrocystis lunula Schütt and unlike that of Alexandrium tamarense (Lebour) Balech et Tangen. The dinoflagellate's own bioluminescence is two orders of magnitude too low to result in self-photoinhibition. The quantitative relationships developed in the laboratory predict photoinhibition of bioluminescence in populations of C. fusus in the North Atlantic Ocean.     

EFFECT OF SEASONAL SEA ICE BREAKOUT ON THE PHOTOSYNTHESIS OF BENTHIC DIATOM MATS AT CASEY,ANTARCTICA1

Photosynthesis of marine benthic diatom mats was examined before and after sea ice breakout at a coastal site in eastern Antarctica (Casey). Before ice breakout the maximum under‐ice irradiance was between 2.5 and 8.2 μmol photons·m^?2·s^?1 and the benthic microalgal community was characterized by low E_k (12.1–32.3 μmol photons·m^?2·s^?1), low relETR_max (9.2–32.9), and high alpha (0.69–1.1). After breakout, 20 days later, the maximum irradiance had increased to between 293 and 840 μmol photons·m^?2·s^?1, E_k had increased by more than an order of magnitude (to 301–395 μmol photons·m^?2·s^?1), relETR_max had increased by more than five times (to 104–251), and alpha decreased by approximately 50% (to 0.42–0.68). During the same time interval the species composition of the mats changed, with a decline in the abundance of Trachyneis aspera (Karsten) Hustedt, Gyrosigma subsalsum Van Heurck, and Thalassiosira gracilis (Karsten) Hustedt and an increase in the abundance of Navicula glaciei Van Heurck. The benthic microalgal mats at Casey showed that species composition and photophysiology changed in response to the sudden natural increase in irradiance. This occurred through both succession shifts in the species composition of the mats and also an ability of individual cells to photoacclimate to the higher irradiances.     

Photosynthesis and transpiration of tomato and CO2 fluxes in a greenhouse under changing environmental conditions in winter

Responses of tomato leaves in a greenhouse to light and CO₂ were examined at the transient stage at the end of winter, when both photoperiod and irradiance gradually increase. Additionally, CO₂ fluxes were calculated for a greenhouse without supplementary lighting and without CO₂ enrichment based on CO₂ sinks (plant photosynthesis) and CO₂ sources (plant and substrate respiration). In January, tomato leaves in the greenhouse showed low photosynthesis with a maximum assimilation of 6–8 μmol CO₂ m⁻² s⁻¹, a quantum yield of 0.06 μmol CO₂ μmol⁻¹ photosynthetic active radiation (PAR) and a low light compensation point of 26 μmol PAR m⁻² s⁻¹, a combination which classifies them as shade leaves. In February, tomato leaves increased their light compensation point to 39 μmol PAR m⁻² s⁻¹ and quantum yield to 0.08, the former indicating the adaptation to increased irradiance and photoperiod. These tomato leaves increased their transpiration from 0.4 to 0.9 in January to ∼2 mmol H₂O m⁻² s⁻¹ in February. Both photosynthesis and transpiration were primarily limited by light but neither by stomatal conductivity nor by CO₂. In January, light response of photosynthesis, dark respiration and transpiration were negligibly affected by increasing CO₂ concentrations from 600 to 900 ppm CO₂ under low light conditions, indicating no benefit of CO₂ enrichment unless light intensity increased. In February, tomato leaves were photoinhibited at inherent greenhouse CO₂ concentrations on the first sunny day; this photoinhibition was further enhanced by an increased CO₂ concentration of 1000 ppm. CO₂ fluxes in the greenhouse appeared strongly dependent on solar radiation. After exceeding the light compensation point in the morning, greenhouse CO₂ concentrations decreased by 58 or by 110 ppm CO₂ h⁻¹ on a sunny day in January or February and by 23 ppm on overcast days in both months. Calculated per overall tomato canopy, plant photosynthesis contributed 42–50% to the morning CO₂ depletion in the greenhouse. Dark respiration of tomato leaves was ∼2 μmol CO₂ m⁻² s⁻¹ in January and ∼3 μmol CO₂ m⁻² s⁻¹ in February. This dark respiration resulted in rises of 15 and 17 ppm CO₂ h⁻¹ at night in the greenhouse compartment and was identified as primary source of CO₂. Respiration of the substrate used to grow the plants, which produced 7.3 ppm CO₂ h⁻¹, was identified as secondary source of CO₂. The combined plant and substrate respiration resulted in peaks of up to 900 ppm CO₂ in the greenhouse before dawn.     

PHYSIOLOGICAL ECOTYPES IN THE MARINE ALGA BOSTRYCHIA RADICANS (CERAMIALES,RHODOPHYTA) FROM THE EAST COAST OF THE U.S.A1

The comparative ecophysiology of nine culture isolates of the eulittoral red alga Bostrychia radicans (Montagne) Montague collected at sites from seven states along the east coast of the U.S.A. was investigated. The growth response in relation to different salinity and light conditions as well as photosynthesis-irradiance curves were studied. In addition, the effect of salt treatment on the content of the isomeric polyols d -sorbitol and d -dulcitol was also studied. All isolates grew between salinities of 5.3 and 70 ppt but with quite different optima and maxima. The isolates were all adapted to low light levels, i.e. growth was already recorded at 2.5 μmol photons·m^?2·s^?1, and growth rates peaked between 40 and 60 μmol photons·m^?2·s_-1. These low-light requirements were also reflected by the photosynthesis-irradiance curves: all plants had low light compensation points (2.5–9.7 μmol photons ·m^?2·^?1) and low photon fluence rates for initial saturation of photosynthesis (38.1–84.7 μmol photons·m^?2·s^?1, indicating that these isolates are “shade-adapted.” Isolates from Florida and Georgia synthesized and accumulated both the osmolytes d -sorbitol and d -dulcitol in increasing salinities, whereas only d -sorbitol was present in plants from North Carolina north to Connecticut. d -sorbitol was always strongly involved in osmotic acclimation. In various isolates from the same location in South Carolina, both polyol patterns were found, i.e. d -sorbitol plus d -dulcitol and d -sorbitol only. All data indicate that B. radicans exhibits a broad salinity tolerance and a low-light preference, which explain the successful colonization of this alga on various intertidal and shaded substrates. The data also clearly indicate intraspecific differences among the nine isolates, which is interpreted as development of different physiological ecotypes.     

太白山红桦种子的萌发特性

 红桦(Betula albo-sinensis)广布于我国太白山中高海拔地区, 普遍存在着更新障碍。为了给红桦林的自然更新提供新的解释与证据, 我们对2 400粒红桦种子进行了去果皮观察并计数发芽率, 观察了红桦种子在不同光合有效辐射(Photosynthetically active radiation, PAR) 和不同光照时间(强光照: PAR为 464.7 μmol photons·m^-2·s^-1, 12 h·d^-1; 中光照: PAR为233.8 μmol photons·m^-2·s^-1, 12 h·d^-1; 弱光 照: PAR为233.8 μmol photons·m^-2·s^-1, 0.5 h·d^-1)、不同昼夜温差(25/20 ℃、20/15 ℃、15/10 ℃)和去果皮处理下的发芽率, 以及无光 照处理下的发芽率。此外, 还观察了种子在不同覆盖物(阔叶、针叶、针阔混合)和不同基质(土、沙)中以及在落叶(阔叶、针阔混合)中的发芽 率。红桦种子成熟时, 大约54.29%为饱满具活力的种子。种子在25/20 ℃的昼夜温差下发芽率最高, 而在15/10 ℃的昼夜温差下极少萌发。中 光照下, 种子发芽率略高于其它两种光照, 无光照则完全抑制了种子萌发。去果皮处理可促进其萌发, 各种覆盖物会导致种子发芽率下降。不 同的萌发基质对萌发无影响。以上结果暗示, 自然条件下太白山红桦种子的萌发将依赖于自然的扰动。     

SHORT‐TERM EFFECT OF TEMPERATURE ON THE PHOTOKINETICS OF MICROALGAE FROM THE SURFACE LAYERS OF ANTARCTIC PACK ICE1

Microalgae growing within brine channels (85 psu salinity) of the surface ice layers of Antarctic pack ice showed considerable photosynthetic tolerance to the extreme environmental condition. Brine microalgae exposed to temperatures above ?5°C and at irradiances up to 350 μmol photons·m^?2·s^?1 showed no photosynthetic damage or limitations. Photosynthesis was limited (but not photoinhibited) when brine microalgae were exposed to ?10°C, provided the irradiance remained under 50 μmol photons·m^?2·s^?1. The highest level of photosynthetic activity (maximum relative electron transport rate [rETR_max]) in brine microalgae growing within the surface layer of sea ice was at approximately 18 μmol electrons·m^?2·s^?1, which occurred at ?1.8°C. Effective quantum yield of PSII and rETR_max of the halotolerant brine microalgae exhibited a temperature‐dependent pattern, where both parameters were higher at ?1.8°C and lower at ?10°C. Relative ETR_max at temperatures above ?5°C were stable across a wide range of irradiance.     

ANTIOXIDATIVE PROTECTION OF PERIDINIUM GATUNENSE IN LAKE KINNERET: SEASONAL AND DAILY VARIATION1

A comprehensive antioxidative mechanism was found in the freshwater dinoflagellate Peridinium gatunense Lemm. during the spring bloom in Lake Kinneret. Ascorbate was present throughout the bloom period and was responsible, together with catalase, for the elimination of photosynthetically produced H₂O₂. As glutathione concentrations and ascorbate regenerative enzymes were negligible during mid-spring, ascorbate was presumably biosynthesized during the photosynthetically active period. Antioxidative activity increased overall at the end of the spring in conjunction with elevated ambient stress conditions, for example high light. Under such circumstances, ascorbate was regenerated. Ascorbate levels doubled when cells were exposed to an increase in irradiance from 60 to 600 μmol photons·m^?2·s^?1, and on addition of H₂O₂, concentrations increased a further 20-fold. Significant antioxidative activity was also noted in the dark, although this was dependent on the presence of H₂O₂. Diurnal changes in antioxidants and their regenerative enzymes were observed. The activities of mono-dehydroascorbate reductase, glutathione reductase, and ascorbate concentrations showed ultraradian periodicity and were completely in phase throughout the day/night period. Dehydroascorbate reductase activity and glutathione concentrations were also in phase but showed aperiodic variation, as did ascorbate peroxidase activity. Superoxide dismutase and catalase activities were generally out of phase during the 24-h period but did show ultraradian periodicity. Lake samples entrained under constant light revealed an inate 12-h rhythm for catalase activity, during at least 36 h.     

Influences of light and UV-B on growth and sporulation of the green alga Ulva pertusa Kjellman

The cosmopolitan presence of Ulva spp. is partly due to its great reproductive ability, but relatively little information is available for the radiation conditions triggering reproduction. In the present study, we investigated the effect of photon irradiance, photoperiod, and spectral qualities of light on growth and reproduction of Ulva pertusa.During 8-day culture of discs cut from marginal parts of the thallus of U. pertusa, the size of the thallus discs was greatest at 10 μmol m⁻² s⁻¹, while saturation of reproduction occurred at 30 μmol m⁻² s⁻¹. The minimum photon irradiance allowing growth and reproduction was 5 and 10 μmol m⁻² s⁻¹, respectively. Rapid increases in the size and subsequent initiation of sporulation were observed in samples transferred to saturating irradiance from 5 μmol m⁻² s⁻¹ or darkness for 9 days. When exposed to different photoperiods (8:16-, 12:12-, 16:8-h LD and continuous light regimes) combined with different photon irradiances (10 and 100 μmol m⁻² s⁻¹), U. pertusa thallus showed that the thallus size attained at the low irradiance was similar in daylengths longer than 12 h per day, while the surface area increased in parallel with increasing light duration at the high irradiance. The degree of sporulation at 10 μmol m⁻² s⁻¹ varied, ranging from no sporulation in 8:16-h LD to 80% in 16:8-h LD and continuous light. On the other hand, there was no remarkable difference in the degree of sporulation between the photoperiods except for slightly smaller percentage sporulation in 8:16-h LD at 100 μmol m⁻² s⁻¹.At 5 μmol m⁻² s⁻¹, the growth of U. pertusa was markedly lower in green than in blue or red light, but there was no sporulation in any spectral quality. The degree of sporulation at 20 μmol m⁻² s⁻¹ was almost twice as much in blue or red as in green light.The size of plants irradiated with 1.0 W m⁻² of UV-B for 20-40 min increased by 18-21% relative to control, whereas higher UV irradiance caused inhibition of growth. There was a significantly lower incidence of sporulation in UV-B-irradiated plants with the degree of reduction being greater in those exposed to higher UV doses. The total biologically effective UV-B dose for 50% inhibition of sporulation was 0.085 Dose_eff kJ m⁻². The time required to achieve 50% inhibition would be longer than 13 h at depths below 1 m in Ahnin coastal waters. The vertical attenuation coefficient of PAR (λ=400-700 nm) and UV-B (λ=300-320 nm) in April 1998 at Ahnin on the eastern coast of Korea was 0.21 m⁻¹ for K_PAR and 0.54 m⁻¹ for K_UV-B. A large reduction of light quantity and rapid disappearance of blue wavelength occurred by shading from overlying thalli.Percentage inhibition of sporulation was only 14-18% at 150-200 μmol m⁻² s⁻¹ compared with 70% at 10 μmol m⁻² s⁻¹, when plants were incubated under different irradiances of PAR immediately after UV-B exposures.These different photoadaptive strategies for sporulation may in part account for the great ecological success of U. pertusa.     

CULTURE OF THE TERRESTRIAL CYANOBACTERIUM,NOSTOC FLAGELLIFORME (CYANOPHYCEAE), UNDER AQUATIC CONDITIONS1

Both colonies and free‐living cells of the terrestrial cyanobacterium, Nostoc flagelliforme (Berk. & Curtis) Bornet & Flahault, were cultured under aquatic conditions to develop the techniques for the cultivation and restoration of this endangered resource. The colonial filaments disintegrated with their sheaths ruptured in about 2 days without any desiccating treatments. Periodic desiccation played an important role in preventing the alga from decomposing, with greater delays to sheath rupture with a higher frequency of exposure to air. The bacterial numbers in the culture treated with seven periods of desiccation per day were about 50% less compared with the cultures without the desiccation treatment. When bacteria in the culture were controlled, the colonial filaments did not disintegrate and maintained the integrity of their sheath for about 20 days even without the desiccation treatments, indicating the importance of desiccation for N. flagelliforme to prevent them from being disintegrated by bacteria. On the other hand, when free‐living cells obtained from crushed colonial filaments were cultured in liquid medium, they developed into single filaments with sheaths, within which multiple filaments were formed later on as a colony. Such colonial filaments were developed at 15, 25, and 30° C at either 20 or 60 μmol photons·m^?2·s^?1; colonies did not develop at 180 μmol photons·m^?2·s^?1, though this light level resulted in the most rapid growth of the cells. Conditions of 60 μmol photons·m^?2·s^?1 and 25° C appeared to result in the best colonial development and faster growth of the sheath‐held colonies of N. flagelliforme when cultured indoor under aquatic conditions.     

COMPARISON OF DEVELOPMENT AND PHOTOSYNTHETIC GROWTH FOR FILAMENT CLUMPS AND REGENERATED MICROPLANTLET CULTURES OF AGARDHIELLA SUBULATA (RHODOPHYTA, GIGARTINALES)

Two axenic, in vitro liquid suspension cultures were established for Agardhiella subulata (C. Agardh) Kraft et Wynne, and their growth characteristics were compared. This study illustrated how reliable routes for the development of suspension cultures of macrophytic red algae of terete thallus morphology can be achieved for biotechnology applications. Undifferentiated filament clumps of 2–8 mm diameter were established by induction of callus-like tissue from thallus explants, and lightly branched microplantlets of 2–10 mm length were established by regeneration of filament clumps. The filament clumps were susceptible to regeneration. Adventitious shoot formation was reliably induced from 40% to 70% of the filament clumps by gentle mixing at 100 rev min^?1 on an orbital shaker. The specific growth rate of the microplantlets was higher than the filament clumps in nonagitated well plate culture (4%–6% per day for microplantlets vs. 2%–3% per day for filament clumps) at 24° C and 8–36 μmol photons·m^?2·s^?1 irradiance (10:14 h LD cycle) when grown on ASP12 artificial seawater medium at pH 8.6–8.9 with 20%–25% per day medium replacement. Oxygen evolution rate vs. irradiance measurements showed that relative to the filament clumps, microplantlets had a higher maximum specific oxygen evolution rate (P_o,max= 0.181 ± 0.035 vs. 0.130 ± 0.023 mmol O₂·g^?1 dry cell mass·h^?1), but comparable respiration rate (Q_o= 0.040 ± 0.013 vs. 0.033 ± 0.017 mmol O₂·g^?1 dry cell mass·h^?1), compensation point (I_c= 3.8 ± 2.4 vs. 5.7 ± 1.2 μmol photons·m^?2·s^?1), and light intensity at 63.2% of saturation (I_k= 17.5 ± 3.9 vs. 14.9 ± 2.6 μmol photons·m^?2·s^?1). The microplantlet culture was more suitable for suspension culture development than the filament clump culture because it was morphologically stable and exhibited higher growth rates.     

MOBILIZATION OF β-1,3-GLUCAN AND BIOSYNTHESIS OF AMINO ACIDS INDUCED BY NH4+ ADDITION TO N-LIMITED CELLS OF THE MARINE DIATOM SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)

Mobilization of the reserve β-1,3-glucan (chrysolaminaran) in N-limited cells of the marine diatom Skeletonema costatum (Grev.) Cleve (Bacillariophyceae) was investigated. The diatom was grown in pH-regulated batch cultures with a 14:10-h light:dark cycle until N depletion. In a pulse-chase experiment, the cells were first incubated in high light (200 μmol photons·m ^{− 2}·s ^{− 1}) with ¹⁴C-bicarbonate until dissolved inorganic carbon was exhausted. Unlabeled bicarbonate (1 mM) was then added, and the cells were incubated in the dark and subsequently in low light (20 μmol photons·m ^{− 2}·s ^{− 1}) with additions of 40 μM NH₄ ⁺ . In the ¹⁴C pulse phase with high light and N depletion, β-1,3-glucan accumulated and accounted for 85% of incorporated ¹⁴C. In the subsequent ¹⁴C chase phases, added NH₄ ⁺ was assimilated at an N-specific rate of 0.11 h ^{− 1} in both the dark and low light, and in both cases it caused a significant mobilization of β-1,3-glucan (dark, 26%; low light, 19%). Biochemical fractionation of organic ¹⁴C showed that free amino acids were most rapidly labeled in the early stage of NH₄ ⁺ assimilation, whereas proteins and polysaccharides were labeled more rapidly after 1.2 h. Analysis of the cellular free amino acids strongly indicated that de novo biosynthesis was occurring, with a Gln:Glu ratio increasing from 0.4 to 10 within 1.2 h. After the NH₄ ⁺ was exhausted, the cellular pools of glucan and amino acids became constant or slowly decreased. In another experiment, N-limited cells were first incubated in high light until dissolved inorganic carbon was exhausted and were further incubated in high light with 150 μM NH₄ ⁺ under inorganic carbon limitation. Added NH₄ ⁺ was assimilated at an N-specific rate of 0.023 h ^{− 1}, and cellular β-1,3-glucan decreased by 15% within 6 h. Hence, β-1,3-glucan was mobilized during NH₄ ⁺ assimilation, even though inorganic carbon was modifying the metabolic rates. The results provide new evidence of β-1,3-glucan supplying essential precursors for biosynthesis of amino acids and other components in S. costatum in both the dark and subsaturating light and even saturating light under inorganic carbon limitation.     

PHOTOSYNTHESIS AND COLD ACCLIMATION: MOLECULAR EVIDENCE FROM A POLAR DIATOM1

The psychrophilic diatom Fragilariopsis cylindrus (Grunow) Krieger in Helmcke & Krieger was used to investigate photosynthesis and growth under freezing temperatures. Gene expression during a temperature shift from +5° C to ?1.8° C was studied under 3 and 35 μmol photons·m^?2·s^?1 by using a macroarray. These measurements were paralleled by determination of fluorescence induction at PSII and pigment analysis. The shift to ?1.8° C at 35 μmol photons·m^?2·s^?1 caused a marginal decrease of photosynthetic quantum yield (F_v/F_m) from 0.61 to 0.52 with fast recovery after 1 day. The ratio of chl c to chl a increased from 3.1 to 5.5, and the ratio of diatoxanthin to diadinoxanthin increased from 0.7 to 5.0. Genes encoding proteins of PSII (psbA, psbC) and for carbon fixation (rbcL) were down‐regulated, whereas genes encoding chaperons (hsp70) and genes for plastid protein synthesis and turnover (elongation factor EfTs, ribosomal protein rpS4, ftsH protease) were up‐regulated. In contrast, cold exposure at 3 μmol photons·m^?2·s^?1 induced a marginal increase in F_v/F_m from 0.61 to 0.63 and a strong increase in fucoxanthin concentrations from 0.04 up to 0.12 pg·cell^?1. This was paralleled by up‐regulation of fcp genes. The ratio of chl c to chl a also increased from 3.1 to 4.2, as did the ratio of diatoxanthin to diadinoxanthin from 0.7 to 2.2. Down‐regulation of psbA, psbC, and rbcL could also be measured but not up‐regulation of hsp70, EfTs, rpS4, and the ftsH protease. The latter genes are probably necessary to avoid cold shock photoinhibition only at higher light intensities.     

Steady-state stomatal responses of C3 and C4 species to blue light fraction: Interactions with CO2 concentration

Blue light induced stomatal opening has been studied by applying a short pulse (~5 to 60 s) of blue light to a background of saturating photosynthetic red photons, but little is known about steady-state stomatal responses. Here we report stomatal responses to blue light at high and low CO₂ concentrations. Steady-state stomatal conductance (g_s) of C₃ plants increased asymptotically with increasing blue light to a maximum at 20% blue (120 μmol m⁻² s⁻¹). This response was consistent from 200 to 800 μmol mol⁻¹ atmospheric CO₂ (C_a). In contrast, blue light induced only a transient stomatal opening (~5 min) in C₄ species above a C_a of 400 μmol mol⁻¹. Steady-state g_s of C₄ plants generally decreased with increasing blue intensity. The net photosynthetic rate of all species decreased above 20% blue because blue photons have lower quantum yield (moles carbon fixed per mole photons absorbed) than red photons. Our findings indicate that photosynthesis, rather than a blue light signal, plays a dominant role in stomatal regulation in C₄ species. Additionally, we found that blue light affected only stomata on the illuminated side of the leaf. Contrary to widely held belief, the blue light-induced stomatal opening minimally enhanced photosynthesis and consistently decreased water use efficiency.     

Identification,biochemical composition and phycobiliproteins production of Chroococcidiopsis sp. from arid environment

Molecular and microscopic studies were performed to identify Chroococcidiopsis sp., an endolithic cyanobacterium, isolated from gypsum rocks of Atacama Desert (Chile). It was adapted to grow in mineral liquid medium, with 9 mM nitrate, bubbled with CO₂-enriched air (2.5 % v/v), and continuously illuminated with a white light of 70 μmol photons m^–2 s^–1. The obtained biomass (productivity of 0.21 g L^–1 d^–1) had a C/N ratio of 6.67, and it contained carbohydrates (45.40 % of dry weight), proteins (36.72 %), lipids (5.60 %) nucleic acids (3.90 %) and ashes (8.28 %). The lipid fraction was particularly rich in palmitic (29.86 % of total fatty acids), linoleic (18.20 %), palmitoleic (12.75 %), linolenic (10.92 %), stearic (9.64 %) and capric acid (6.29 %). Chroococcidiopsis sp. accumulated phycobiliproteins in a light-dependent process and produced 204 mg g^–1, under incident light of 10 μmol photons·m^–2·s^–1, with a relative abundance of 40.9 % for phycocyanin, 23.3 % for phycoerythrin, and 35.8 % for allophycocyanin. The biomass from this cyanobacterium can be a good source of these pigments, especially APC (maximum of 95 mg g dw⁻¹), which are of interest for pharmacological, cosmetic, and food industries.     

Effect of light intensity and eye development on prey capture by larval striped bass Morone saxatilis

The efficacy of visual and non-visual feeding among pelagic striped bass Morone saxatilis larvae adapted to a turbid estuary was determined in the laboratory in clear water. Capture of Artemia salina (density 100 l⁻¹) was significantly affected by the interaction between age of larvae (range: 8–25 days post-hatch, dph) and light intensity (range: 0–10·6 μmol s⁻¹ m⁻² at the water surface). Visual feeding by larvae aged 9–11 dph was highest in dim light (0·086–0·79 μmol s⁻¹ m⁻²), with fish capturing up to 5 prey larva⁻¹ h⁻¹. As the larvae grew, prey capture in brighter light improved, associated with an increasing proportion of twin cone photoreceptors and improving ability of the retina to light- and dark-adapt. By age >22 dph, mean prey capture was greatest at highest light intensities (0·79 and 10·6 μmol s⁻¹ m⁻²) exceeding 100 prey larva⁻¹ h⁻¹. Incidence of feeding larvae generally improved as the larvae grew, reaching >80% in all light intensities from 16 dph onwards. The lower threshold for visual feeding, between 0·0084 and 0·03 μmol s⁻¹ m⁻², remained constant as the larvae grew, despite an increasing density of rod photoreceptors. Below this threshold, non-visual feeding was evident at a low rate (<6 prey larva⁻¹ h⁻¹) that was independent of larval age.