Studies on 14CO2-assimilation in marine rhodophyceae

Experiments on enzymatic (in vitro) and in vivo CO₂-fixation using a variety of marine Rhodophyceae such as Porphyra umbilicalis (L.) J.Ag., Rhodomela confervoides (Huds.) Silva, Corallina officinalis L. and Chondrus crispus Stackh. revealed that carbon assimilation in the Rhodophyceae is almost exclusively performed by photosynthesis via ribulose-1, 5-di-phosphate carboxylase, whereas light-independent CO₂-fixation via -carboxylation by phosphoenolpyruvate carboxykinase scarcely exceeds 1% of the total carbon fixation potential of the plants. Activity of phosphoenolpyruvate carboxylase could not be detected. With respect to the main accumulation products of photosynthetic CO₂-fixation, the Rhodophyceae investigated are not uniform: Corallina officinalis L., Rhodymenia palmata (L.) Grev., and Gigartina stellata (Stackh.) Batt. have been found to accumulate ¹⁴C in the neutral compound floridoside (=2-O-glycerol--D-galactopyranose), whereas Delesseria sanguinea (Huds.) Lamour., Ceramium cubrum (Huds,) C.Ag., and Rhodomela confervoides (Huds.) Silva, representing members of the Delesseriaceae, Ceramiaceae and Rhodomelaceae, respectively, do not photosynthesize floridoside, but show intense ¹⁴C-labelling in an acidic constituent, mannosidoglycerate (= digeneaside). This compound is reported for the first time as a rapidly ¹⁴C-labelled and accumulated photosynthate in a variety of red algal species exclusively belonging to the Ceramiales.     

Settlement of larval blacklip abalone, Haliotis rubra, in response to green and red macroalgae

Surfaces from the habitat of adult Haliotis rubra were tested as inducers of larval settlement to determine the cues that larvae may respond to in the field. Settlement was high on the green algal species Ulva australis and Ulva compressa (Chlorophyta), the articulated coralline algae Amphiroa anceps and Corallina officinalis, and encrusting coralline algae (Rhodophyta). Biofilmed abiotic surfaces such as rocks, sand and shells did not induce settlement. Ulvella lens was also included as a control. Treatment of U. australis, A. anceps and C. officinalis with antibiotics to reduce bacterial films on the surface did not reduce the settlement response of H. rubra larvae. Similarly, treatment of these species and encrusting coralline algae with germanium dioxide to reduce diatom growth did not significantly reduce larval settlement. These results suggest that macroalgae, particularly green algal species, may play an important role in the recruitment of H. rubra larvae in the field and can be used to induce larval settlement in hatchery culture.     

Effect of diurnal variations in natural irradiance on the apical length growth and light saturation of growth in five species of benthic macroalgae

The diurnal variation in apical length growth rate of five benthic macroalgae from the Swedish West Coast (Fucus spiralis L., F. vesiculosus L., F. serratus L., Ascophyllum nodosum (L.) Le Jolis, and Chondrus crispus Stackhouse) was measured during 1.5 to 2.0-h intervals for 3 to 5 d in September 1984. All species showed a maximum in growth rate in the morning, followed by a continuous decline throughout the rest of the day (most pronounced in F. serratus, A. nodosum and C. crispus). By comparing the growth rates with the average irradiance during the same interval, saturation irradiances for length growth in natural daylight regime were estimated; 60 to 70 Wm^-2 for F. spiralis and F. vesiculosus, 90 to 100 Wm^-2 for A. nodosum, 30 to 40 Wm^-2 for F. serratus and 10 to 20 Wm^-2 for C. crispus. Average growth rates during the dark hours ranged from 33% of average day growth rates in A. nodosum to 63% in C. crispus, respectively.     

Feeding and nutritional ecology of the sea urchin strongylocentrotus drobachiensis in Maine,USA

The sea urchin strongylocentrotus drobachiensis exhibited a high degree of food selectivity, whether foods were presented singly or in combination. Foods ranked from most to least preferred were, in summer, Laminaria longicruris, Chondrus crispus, Corallina officinalis, Ascophyllum nodosum, and Agarum cribrosum, whereas in winter A. nodosum and A. cribrosum exchanged ranks. Food preference was not correlated with caloric content but, because of higher feeding rates on preferred foods, caloric intake was positively correlated with preference. Similarly, food absorption rankings were not correlated with food preference, with the exception of gravimetric efficiencies in winter. However, the absorption of L. longicruris, the most preferred alga, was highest in all measurements. Growth and reproductive development of S. drobachiensis on single species diets were positively correlated with food preference. Highest values occurred with animals fed L. longicruris. Urchins transferred from non-preferred to preferred diets showed increased somatic and reproductive growth compared to control animals on the original, non-preferred diets. Conversely, when transferred from preferred to non-preferred diets, urchins showed reduced growth compared to controls. No combination diet tested supported significantly better gonadal growth than L. longicruris: 25% C. crispus supported slightly better somatic growth than L. longicruris alone. These data support the view that S. drobachiensis has evolved a feeding strategy resulting in the maximization of growth and reproduction.     

Changes in photosynthetic pigment concentration in seaweeds as a function of water depth

We conducted a study of the relationship between changes in photosynthetic pigment content and water depth in Great Harbor near Woods Hole, Massachusetts, USA, on the green algae Ulva lactuca and Codium fragile and the red algae Porphyra umbilicalis and Chondrus crispus. A calibrated underwater photometer equipped with spectral band filters measured light attenuation by the water column. The depth required for a 10-fold diminution of photon flux was 3.6, 5.3, 6.0 and 6.0 m for red, blue, yellow and green light, respectively. Seaweeds were attached to vertically buoyed lines and left to adapt for 7 days; then, with their positions reversed, they were allowed to readapt for 7 days. All species showed greater photosynthetic pigment content with increased depth. Further, the ratio of phycobiliproteins and chlorophyll b to chlorophyll a increased with depth. Changes in pigment content were reversible and occurred in the absence of cell division. There was a net loss of pigments near the surface (high irradiance), and subsequent synthesis when seaweeds were transferred to a position deep in the water column (low irradiance). In contrast, seaweeds which were found in intertidal habitats changed only their pigment concentration, and not pigment ratio, a phenomena analogous to higher plant sun and shade adaptation. Therefore, seaweeds modify their photon-gathering photosynthetic antennae to ambient light fields in the water column by both intensity adaptation and complementary chromatic adaptation.     

Photosynthetic studies of Chondrus crispus

The net photosynthesis of intertidal, subtidal, carposporic, tetrasporic, and winter versus summer acclimatized plants of Chondrus crispus Stackhouse were evaluated under different temperatures and quantities of light. The optimum temperature and light conditions for net photosynthesis of C. crispus are seasonally and spatially variable, and there is an adaptive shift in the photosynthetic capacity at different seasons and positions on the shore. Plants collected during the fall and winter had lower light optima (465 to 747 ft-c) for net photosynthesis than spring and summer specimens (about 1000 ft-c). Intertidal populations exhibited a higher rate of net photosynthesis between 250 and 2819 ft-c than subtidal plants. Summer materials have a greater tolerance to high temperatures and a higher temperature optimum than winter materials. Shallow subtidal populations (-6m) exhibited a higher temperature optimum than deep subtidal plants (-12m). Tetrasporic plants (diploid) showed a higher rate of net photosynthesis than carposporic plants (haploid). It is suggested that the diploid plants of C. crispus may extend deeper in the subtidal zone, because they have a higher rate of net photosynthesis than carposporic plants. The results of the present studies are compared with previous physiological studies of C. crispus.Published with the approval of the Director of the New Hampshire Agriculture Experiment Station as Scientific Contribution Number 742.     

Effect of freezing on photosynthesis of intertidal macroalgae: relative tolerance of Chondrus crispus and Mastocarpus stellatus (Rhodophyta)

The effect of freezing on photosynthetic metabolism was studied in the red algae, Chondrus crispus and Mastocarpus stellatus. Plants of both species were collected from the intertidal at Chamberlain or Kresge Point, Maine, USA (43°56N, 69°54W) between February and March 1987. Photosynthetic rates were measured immediately after freezing at-20°C and following recovery periods in seawater. Photosynthesis in C. crispus declined rapidly following freezing, falling to 70% of control values within 1 h and 30% after 3 h exposure. Minimum photosynthetic rates (7 to 9% of controls) occurred following freezing exposures of 12 h or more. Full photosynthetic recovery in C. crispus after 3 h at-20°C required 48 h. Photosynthesis in C. crispus did not fully recover in plants frozen for 6 h or more. In contrast, photosynthesis in M. stellatus was relatively unaffected by freezing exposures of <12 h. Twelve hours or more at-20°C reduced photosynthesis to 55% of controls. Photosynthesis in M. stellatus fully recovered from 24 h at-20°C within 24 h. In both species the reduction of photosynthesis by freezing was associated with damage to the plasma membrane and reduced efficiency of energy transfer from phycobilisomes to chlorophyll a, but did not appear to involve ribulose-1,5-bisphosphate carboxylase oxygenase activity. The freezing tolerance of C. crispus and M. stellatus positively correlates with their respective intertidal distributions, suggesting that freezing may be involved in controlling the distributions of these species on the shore.     

Distinguishing modes of dissolved organic carbon production in marine macrophytes by tracer kinetic analysis

Tracer kinetic analysis of radioisotope incorporation into dissolved organic compounds reveals two distinct patterns of photosynthate release by macroalgae. In experiments employing Sargassum lacerifolium, dissolved organic carbon was produced at a constant rate during light incubations. Steady state rates of production were never achieved in experiments employing either Ecklonia radiata (Turn.) J. Agardh. or Ulva lactuca L. Analysis of the time-varying radioactivity curves obtained in experiments using these algae always resulted in models consistent with dissolved organic carbon production being an autocatalytic process. Preincubation of U. lactuca in the dark resulted in a diminished (ca. 40%) rate of dissolved organic carbon production during the subsequent light incubations. In no case did the radioisotope content of the dissolved organic carbon approach a limiting value, indicating that in contrast to phytoplankton, uptake rates of photosynthate by macroalgae are always less than the rates of production.     

Freezing tolerance in the intertidal red algaeChondrus crispus andMastocarpus stellatus: Relative importance of acclimation and adaptation

The effect of repeated daily freezing on photosynthesis, growth and phenotypic acclimation to freezing was studied in the red algaeChondrus crispus Stackhouse andMastocarpus stellatus (Stackhouse in With.) Guiry. Algae used for experiments were collected from Chamberlain, Maine, between March and August 1987, and field observations and experiments were carried out at Chamberlain and Kresge Point, Maine between March 1987 and March 1989. After ca 30 d of daily freezing for 3 h at –5°C photosynthesis ofC. crispus was reduced to 55% of control values. Growth rates ofC. crispus were also reduced in fronds frozen daily compared to unfrozen controls, and eventually fronds became bleached and fragmented resulting in biomass losses. Fronds ofC. crispus, frozen daily, had higher photosynthetic rates following freezing events than unfrozen controls indicating that this species can acclimate to freezing conditions. Acclimation to freezing involves the light-harvesting reactions of photosynthesis. In contrast, photosynthesis and growth inM. stellatus were unaffected by repeated daily freezing for 3 h at –5°C for 36 d. No differences in photosynthesis following freezing were observed between frozen and control fronds suggesting thatM. stellatus does not phenotypically acclimate to freezing. The greater freezing tolerance ofM. stellatus relative toC. crispus results, in part, from genetic adaptations associated with plasma membranes and the light-harvesting reactions of photosynthesis.     

Estimating variation in surface emissivities of intertidal macroalgae using an infrared thermometer and the effects on temperature measurements

Accurate measurements of surface temperatures with an infrared (IR) thermometer require input of the emissivities of the surfaces being measured; however, few determinations of the emissivities of intertidal organisms’ surfaces have been made. Emissivities of intertidal macroalgae were measured to determine whether algal species, measurement angle, hydration, and layering affected them. Emissivities were similar and averaged 0.94 among 11 of 13 species. The species with lower and more variable emissivities (Chondracanthus exasperatus and Desmarestia viridis) differed in morphology from the other species, which were relatively flat thin blades with little surface texture. Measurement angle caused emissivities to decrease significantly in Mazzaella splendens but not in three other species. Hydration and layering of Ulva lactuca also had no effect. At 22 °C, measured temperatures were within 1 °C of actual temperatures when thermometer emissivity settings ranged from 0.75 to 1.00. When emissivities were set lower than actual values, measured temperatures were lower than actual temperatures at 15 °C and higher than actual temperatures at 60 °C. When the IR thermometer was used to measure surface temperatures of nine species of intertidal algae immediately before they were inundated by the incoming tide, temperatures were higher in mid intertidal than low intertidal individuals and higher on a sunnier day than an overcast day. Temperatures of U. lactuca increased with increasing height on the shore, but temperatures of Ulvaria obscura did not. Temperatures were also higher in Fucus distichus blades than receptacles, and lower in U. lactuca and M. splendens occurring in the lower layers of stacks of algae.     

Temperature-influenced infection rates in the Chondrus crispus—Petersenia pollagaster pathosystem: a regression analysis

Cross-infection experiments were performed to determine the influence of temperature on infection rate in the Chondrus crispus Stackhouse-Petersenia pollagaster (Petersen) Sparrow pathosystem. C. crispus thalli were collected at Pubnico Harbor, Nova Scotia, Canada in the fall of 1981 to 1984. Infective zoospores were used to inoculate healthy thalli at five different temperatures. The highest infection rate was obtained at 20°C, while significantly lower rates were obtained at temperature extremes. The parasite's life cycle, consisting of infection of healthy thalli, endobiotic development, and release of zoospores, was completed in 48 to 72 h at 15° to 20°C.     

Inhibitory effects of red algal extracts on larval settlement of the barnacle Balanus improvisus

We examined the chemical antifouling properties of four sublittoral red algae, Chondrus crispus, Delesseria sanguinea, Osmundea ramosissima, and Polyides rotundus, which are all rarely fouled in the field. Two different approaches were used. Firstly, we tested the effects of lipophilic crude extracts on the settlement behaviour of cyprid larvae of the co-existing barnacle Balanus improvisus. Secondly, in a settlement preference experiment, we tested whether B. improvisus cyprid larvae settle on living algae when given a choice between natural algal surfaces and control surfaces. With this procedure, we were able to test both if the algae inhibit recruitment of cyprids, and if this inhibition is a result of chemistry. The settlement of B. improvisus larvae was strongly inhibited at concentrations estimated to be potentially ecologically relevant for all of the tested extracts. However, only C. crispus significantly inhibited settlement in the preference experiment, even though there was also a tendency for settlement inhibition on P. rotundus and O. ramosissima. In contrast, D. sanguinea seemed to stimulate settlement. This contradiction probably resulted from an extraction of metabolites that naturally occur only inside the alga. However, as this study shows, a combination of settlement assays with whole-cell extracts and preference tests of ecologically relevant fouling organisms on natural algal and control surfaces may be a useful procedure to avoid erroneous conclusions regarding natural antifouling roles of compounds based on settlement assays with only whole-cell extracts. Furthermore, this study also shows that production of inhibitory metabolites may explain the low degree of fouling, especially by B. improvisus, on C. crispus.Communicated by L. Hagerman, Helsingør     

Ecological energetics of the seaweed zone in a marine bay on the Atlantic coast of Canada. I. Zonation and biomass of seaweeds

An intertidal and underwater survey of the zonation of seaweed in St. Margaret's Bay, NovaScotia, Canada showed 8 major zones as one proceeded away from shore: (1) Fucus and Ascophyllum; (2) Chorda with filamentous browns; (3) Chondrus crispus; (4) Zostera marina; (5) Laminaria digitata with L. longicrusis; (6) Laminaria longicruris; (7) L. longicruris with Agarum cribrosum; (8) Agarum cribrosum with Ptilota serrata. Zostera occurred at the same level as O. crispus but replaced it in sheltered water. Ascophyllum was more abundant in sheltered water. L. digitata was confined to the more exposed, steeper shore. The average horizontal extent of the seaweed zone was 369 m, and the greatest depth of significant amounts of seaweed 20 to 30 m. Laminaria longicruris contributed 36% of the total biomass, and Laminaria spp. and Agarum together constituted 83%, while intertidal seaweeds contributed less than 10% of the biomass. The estimated average total biomass per m of shore line was 1,481 kg fresh weight, 326 kg dry weight, 98 kg carbon, or 980x10³ kcal. When averaged over the whole area of the bay, the corresponding figures were 1.38 kg/m² fresh weight, 0.30 kg/m² dry weight, 91 g/m² carbon or 912 kcal/m². Dry matter of Laminaria was 15 to 27% of fresh weight in blades, 10 to 12% in stipes. The dry matter content of blades was least in spring and highest in autumn, but carbon content and calorific value of dry matter showed little difference with species or season.Contribution to the International Biological Programme CCIBP 108.Bedford Institute Contribution BI 249.     

Comparative feeding preference of Strongylocentrotus droebachiensis (Echinoidea) for the invasive seaweed Codium fragile ssp. tomentosoides (Chlorophyceae) and four other seaweeds

Laboratory experiments conducted during 1987 on Appledore Island, Maine, USA, tested whether feeding preference or the absence of an attractant was the cause for the occurrence of beds of Codium fragile ssp. tomentosoides (herein referred to as Codium fragile) within rocky barrens grazed clear of kelp by the sea urchin Strongylocentrotus droebachiensis. Consumption of C. fragile in single-diet experiments (1 seaweed/sea urchin) was highly variable and was not significantly different from that for several other seaweeds (Agarum cribrosum, Ascophyllum nodosum, Chondrus crispus, and Laminana saccharina) important in the field diet of the green sea urchin. In multiple-diet experiments (5 seaweeds/sea urchin) significantly less Codium fragile was eaten than Chondrus crispus, but significantly more Codium fragile was eaten than A. cribrosum. Chemosensory experiments suggest that C. fragile does not attract the sea urchin. Sea urchins are unable to detect C. fragile but will eat it when they come in contact with it.     

Seasonal variations in salt-march macroalgae photosynthesis. II. Fucus vesiculosus and Ulva lactuca

Photosynthesis in whole plants of the salt-marsh algae Fucus vesiculosus and Ulva lactuca was evaluated by ¹⁴C-uptake under a variety of light intensities at approximately mately monthly intervals during a 15-month study. Photosynthetic capacity in both species was closely related to seasonal irradiation patterns and changes in field biomass. Maximum photosynthesis occurred in the spring and summer months. Photosynthesis on a dry weight basis was higher in U. lactuca, while photosynthesis on a chlorophyll a basis was equal in both species. Photosynthetic capacity was inversely related to pigment content. Maximum chlorophyll a concentrations occurred during the winter. Frond profile studies in F. vesiculosus indicated that apices always exhibited greatest photosynthetic capacity. Uptake of ¹⁴C into ethanol-soluble and insoluble fractions was different in each species. F. vesiculosus showed greater activity in the ethanol-soluble fraction while U. lactuca exhibited greater activity in ethanol-insoluble fractions.This research was supported by Research Grant AG-375 from the National Science Foundation and, in part, by the State University of New York Research Foundation and the Energy Research and Development Administration (ERDA).     

Nitrate reductase and glutamine synthetase activity,internal N pools,and growth of<Emphasis Type="Italic"> Ulva lactuca</Emphasis>: responses to long and short-term N supply

Fast-growing macroalgae, including Ulva lactuca Linnaeus, respond rapidly to changes in nutrient conditions, particularly to short-term N supply. This ability to rapidly take up and assimilate N contributes to the increasing occurrence of macroalgal blooms in heavily N loaded coastal ecosystems. To determine whether long-term nutrient histories affect short-term responses in activity of N-assimilating enzymes, including nitrate reductase (NRA) and glutamine synthetase activity (GSA), internal N storage, and macroalgal growth, we conducted an in situ nitrate fertilization experiment between 7 and 22 July 2004, with fronds of U. lactuca collected from estuaries with high and low N loads in Waquoit Bay, Cape Cod, Massachusetts, USA (N 41° and W 70°). Initial NRA, GSA, % N, δ¹⁵N, and growth of U. lactuca fronds were higher in the site where nitrate was in high supply. Differences in NRA persisted even after short-term experimental enrichment. Differences in internal N pools, δ¹⁵N, and growth, in contrast, mirrored the changes of nutrient supply. The rate of turnover of the internal N content of U. lactuca was quite short (<2 d), and turnover of enzyme activity may have been even shorter. N isotopic fractionation by U. lactuca appeared to be of small magnitude, unlike the case of phytoplankton, and similar to that of vascular plants. δ¹⁵N was a better indicator of short-term response to external and internal nutrient supplies in U. lactuca than enzyme activity or N content, and may reliably detect rapid changes in N availability, source, and uptake and assimilation processes.     

Rapid growth of clones of the red alga Chondrus crispus: applications in assays of toxic substances and in physiological studies

Growth rates of excised apical segments from three Chondrus crispus Stackhouse clones were rapid, reproducible and easily measured using simple equipment. Clonal segments exhibited a high degree of phenotypic stability, with coefficients of variation in growth rates being typically less than 5%. Sensitivity of the assay was demonstrated using 24-h pulses of the toxicants Cu²⁺ (10 to 150 ppb) and the molluscicide Bayluscide (100 to 500 ppb). As a further demonstration of applicability of this assay system, growth rates were measured over two consecutive 24-h photoperiods using a normally pigmented red gametophytic clone and a green colored mutant derived from it. Growth rates were highest in the first hours of the light phase for both clones, with growth of the green mutant being consistently inferior to that of the normally pigmented one. Significant changes were measured with intervals as short as 4 h using only five apical segments for each treatment. The C. crispus assay described is sensitive, relatively rapid, and statistically robust when applied to toxicant testing in seawater and to physiological studies.     

Fluorescence induction in the macroalgae Chondrus crispus (Rhodophyceae) and Ulva sp. (Chlorophyceae)

The induction of in vivo chlorophyll a (chl a) fluorescence (change in fluorescence intensity during a time-scale of ms to s) was measured to determine the potential of this technique for assessing the physiological condition of the macroalgae Chondrus crispus and Ulva sp. A gradient in variable fluorescence (P-F ₀ =peak minus initial fluorescence, a measure of Photosystem II activity) was found along the frond of C. crispus, the values increasing with distance from the thallus apex. No gradient was observed for Ulva sp. thalli. Nitrogen- or phosphorus-depleted Ulva sp. required a longer dark-conditioning period and had lower values of P-F ₀ than did controls. In contrast, no differences were found in P-F ₀ of N- or P-depleted C. crispus unless values were normalized to chl a. The irradiance history of C. crispus strongly influenced P-F ₀ , even after dark-conditioning: P-F ₀ declined by about 70% as the mean daily natural irradiance increased between 2 February and 14 March 1986; a negative correlation was observed between P-F ₀ and the photon flux density 1 d prior to the measurement; P-F ₀ remained elevated when C. crispus was grown under a low photon flux density; and P-F ₀ decreased in thalli within 5 d of transfer from growth under natural irradiance to an incubator with artificial irradiance. Changes in variable fluorescence at different growth irradiances of C. crispus may be due to adaptive changes in the relative absorption cross-section of this alga. The influence of irradiance history on Ulva sp. was minimal in comparison.Issued as NRCC No. 28730Part of this study was carried out while employed by Focal Marine Ltd., Bedford, Nova Scotia, Canada     

Kinetic measurements of metal accumulation in two marine macroalgae

 We measured the uptake kinetics of four metals (Cd, Cr, Se and Zn) in two marine macroalgae (the green alga Ulva lactuca and the red alga Gracilaria blodgettii). Metal uptake generally displayed a linear pattern with increasing exposure time. With the exception of Cr, which exhibited comparable uptake rate constants at different concentrations, uptake rate constants of Cd, Se and Zn decreased with increasing metal concentration, indicating that the seaweeds had a higher relative uptake at lower metal concentration. Uptake of Cd and Zn was higher in U. lactuca than in G. blodgettii, whereas uptake of Cr and Se was comparable between the two species. Only Cd and Zn uptake in U. lactuca was significantly inhibited by dark exposure. A decrease in salinity from 28 to 10‰ enhanced the uptake of Cd, Cr, Se and Zn in U. lactuca 1.9-, 3.0-, 3.6-, and 1.9-fold, respectively. In G. blodgettii, Cd uptake increased twofold when salinity was decreased from 28 to 10‰, whereas uptake of Cr and Zn was not significantly affected by salinity change. The calculated depuration rate constants of metals in U. lactuca were 0.01 d⁻¹ for Cd, 0.05 to 0.08 d⁻¹ for Cr, 0.14 to 0.16 d⁻¹ for Se, and 0.12 to 0.15 d⁻¹ for Zn, and were relatively independent of the metal body burden in the algae. The predicted bioconcentration factor was 3 × 10⁴ for Cd, 2 × 10³ for Cr, 40 to 150 for Se, and 1 to 2 × 10⁴ for Zn in U. lactuca. Our kinetic study suggested that U. lactuca would be a good biomonitor of Cr and Zn contamination in coastal waters. Received: 14 September 1998 / Accepted: 29 May 1999     

Inorganic carbon uptake by photosynthetically active protoplasts of the red macroalga Chondrus crispus

Photosynthetically active protoplasts were isolated from Chondrus crispus Stackh. by treating thalli with -carrageenase produced from batch culture of Pseudomonas carrageenovora. Using the silicone oil centrifugation technique, it was found that the protoplasts: (1) did not generally accumulate inorganic carbon (C_i) above the concentration in their incubation medium; (2) were saturated at C_i concentrations of 3 to 4 mM; (3) had an intracellular pH of 7.50 when incubated at pH 7.5; and (4) their initial carbon fixation rate was reduced by carbonic anhydrase inhibitors. Although the carbon fixation rate of the protoplasts was about 30% that of thallus fragments, presumably due to the relatively harsh protoplast isolation treatment, the behavior of the protoplasts was similar to that of fragments. This similarity indicates that the protoplasts are photosynthetically active and behave as thallus fragments. Further, the data are consistent with the hypothesis that C. crispus acquires C_i for photosynthesis by the diffusion of CO₂ across the plasma membrane.