OCCURRENCE OF CODIUM FRAGILE SUBSP. TOMENTOSOIDES (CHLOROPHYTA: BRYOPSIDALES) IN MARINE EMBAYMENTS OF SOUTHEASTERN AUSTRALIA

Codium fragile (Sur.) Hariot subsp. tomentosoides (van Goor) Silva, a widespread weedy species, is reported from Australia for the first time. Specimens were initially found in June 1996 in Corner Inlet, Victoria, where the species fouled fishing nets. This invasive chlorophycean alga was subsequently found in large numbers on 30 January 1997 in Port Phillip Bay and on 9 March 1998 in Western Port, Victoria. Presumably the species is native to Japan. The alga formed dense stands on rocky substrata in intertidal or in subtidal regions. In Port Phillip Bay, the species was also found attached to the common blue mussel Mytilus edulis planulatus Lamarck, and in Western Port it was found attached to the mud oyster Ostrea angasi Sowerby. Results of this study suggest that recruitment of juvenile germlings and growth of C. fragile subsp. tomentosoides differs between populations in Port Phillip Bay and those in Western Port.     

EFFECT OF INDOLE-3-ACETIC ACID ON GROWTH OF CODIUM FRAGILE SUBSP. TOMENTOSOIDES (CHLOROPHYCEAE) IN CULTURE1

Growth of Codium fragile subsp. tomentosoides (van Goor) Silva in culture depends upon the season of seawater collection. One factor responsible for this variation in growth may be indole-3-acetic acid (IAA). When 10^?9 to 10^?4 M IAA is added to cultures of Codium fragile, optimum growth is at 10^?6 M. The response to exogenous IAA depends upon the time of year when the sea-water is collected. The growth in a range of known IAA concentrations allows the prediction of a seasonal cycle of IAA, or its physiological equivalent, in Rhode Island coastal waters. Such a compound may be an important ecological factor for some algal species.     

CHANGES IN INTRACELLULAR NITROGEN POOLS AND FEEDBACK CONTROLS ON NITROGEN UPTAKE IN CHAETOMORPHA LINUM (CHLOROPHYTA)1

Changes in the size of intracellular nitrogen pools and the potential feedback by these pools on maximum N uptake (NH₄⁺ and NO₃^?) rates were determined for Chaetomorpha linum (Müller) Kützing grown sequentially under nutrient-saturating and nutrient-limiting conditions. The size of individual pools in N-sufficient algae could be ranked as residual organic N (RON) comprised mainly of amino acids and amino compounds > protein N > NO₃^? > NH₄⁺ > chlorophyll N. When the external N supply was removed, growth rates remained high and individual N pools were depleted at exponential rates that reflected both dilution of existing pools by the addition of new biomass from growth and movement between the pools. Calculated fluxes between the tissue N pools showed that the protein pool increased throughout the N depletion period and thus did not serve a storage function. RON was the largest storage reserve; nitrate was the second largest, but more temporary, storage pool that was depleted within 10 days. Upon N resupply, the RON pool increased 3 × faster than either the inorganic or protein pools, suggesting that protein synthesis was the rate-limiting step in N assimilation and caused a buildup of intermediate storage compounds. Maximum uptake rates for both NH₄⁺ and NO₃^? varied inversely with macroalgal N status and appeared to be controlled by changes in small intracellular N pools. Uptake of NO₃^? showed an initial lag phase, but the initial uptake of NH₄⁺ was enhanced and was present only when the intracellular NH₄⁺ pool was depleted in the absence of an external N supply. A strong negative correlation between the RON pool size and maximum assimilation uptake rates for both NH₄⁺ and NO₃^? suggested a feedback control on assimilation uptake by the buildup and depletion of organic compounds. Enhanced uptake and the accumulation of N as simple organic compounds or nitrate both provide a temporary mechanism to buffer against the asynchrony of N supply and demand in C. linum.     

NITROGEN UPTAKE BY FUCUS SPIRALIS (PHAEOPHYCEAE)1,2

Ammoniun, nitrate and nitrite update by Fucus spiralis L. from the Massachusetts coast was examined. Uptake of all appeared to follow saturation type nutrient uptake kinetics, with uptake often restricted at ambient nutrient concentrations. Although only relatively large difference in K₈ values could be easily distinguished, K₈ values for NO_3? and NH₄₊ were generally similar and low compared with NO_2?. There was also some suggestion that K₈ was reduced at lower temperatures. At 15 C. V_max for light and dark uptake for both NH₄+ and NO_3?, and light uptake of N0_2? were similar, suggesting comparable potential use at higher concentrations. Ammonium and NO_3?uptake decreased at lower temperatures giving Q_ro values of 1.8 and 1.6, respectively, between 5 and 15°C. Nitrate and NH₄+ were taken up together and high levels of NH₄+ did not inhibit NO_3? uptake. Light did not affect uptake of either but did stimulate NO_2? uptake. Ammonium and NO_3? uptake were highest in apical frond and whole young plants, and lowest in slower growing, older frond and stipe. On a relative basis. NO_3?, NH₄₊ and NO_2? were estimated to have contributed ca. 59, 39 and 2% respectively, to the yearly N uptake by apical frond. During winter, NO_3? would provide ca. twice the N to F. spiralis as would, NH₄+. From summer to early fall, when NO_3? levels are lower, NO_3? and NH₄+ would be used in comparable amounts.     

INTRON REVEALED BY NUCLEOTIDE SEQUENCE OF LARGE SUBUNIT OF RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE/OXYGENASE FROM CODIUM FRAGILE (CHLOROPHYTA): PHYLOGENETIC ANALYSIS1

The coding sequence for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) from Codium fragile (Suringar) Hariot chloroplast DNA is 1428 bp in length and contains a 1813-bp group II intron. The only other organisms in which introns have been found in the rbcL gene are Euglena and Astasia. The Codium intron likely had a separate origin from the Euglena and Astasia introns, based on comparisons of intron sizes and sequences. Phylogenetic analyses of rbcL nucleotide and amino acid sequences place Codium between Chlorella and two Chlamydomonas spp., indicating that the Chlorophyceae may be polyphyletic.     

INTERSPECIFIC VARIATION IN DARK NITROGEN UPTAKE BY DINOFLAGELLATES1

Seven species of marine dinoflagellates were grown in nitrogen-sufficient media under a 12:12 h L:D cycle, and then tested for their ability to take up nitrate and ammonium in the light and in the dark in short-term experiments with ¹⁵N-labelled substrate. The effect of the N substrate chosen, and the effect of sampling time in the L:D cycle, on the relative nitrogen content (the C:N ratio) was investigated at the same time. The physiological extremes in the material were represented by Prorocentrum minimum (Pav.) J. Schiller, which took up and presumably assimilated nitrate equally fast in the light and in the dark, and Gyrodinium aureolum Hulburt, which did not take up nitrate in the dark when in a state of nitrogen sufficiency. A strong coupling between nitrate assimilation and photosynthetic carbon assimilation in the latter species was suggested by the close similarity of the light saturation curves of ¹⁵NO₃^? and ¹⁴CO₂ incorporation, and by a complete blocking of ¹⁵NO₃^? incorporation by 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Nitrogen starvation for 24 h induced a capacity in G. aureolum for taking up nitrate in the dark, or in the light in the presence of DCMU, a phenomenon that might be useful for identifying nitrogen limitation in this species in the field. Our study emphasizes the variability of dinoflagellate nitrogen nutrition and illustrates the difficulty of associating mass occurrences of dinoflagellates in nature with any particular nutritional mode.     

METABOLIC REGULATION OF AMMONIUM UPTAKE BY ULVA RIGIDA (CHLOROPHYTA): A COMPARTMENTAL ANALYSIS OF THE RATE-LIMITING STEP FOR UPTAKE1

Non-linear time courses of ammonium (NH₄⁺) depletion from the medium and internal accumulation of soluble nitrogen (N) in macroalgae imply that the rate-limiting step for ammonium uptake changes over time. We tested this hypothesis by measuring the time course of N accumulation in N-limited Ulva rigida C. Agardh. Total uptake was measured as removal of NH₄⁺ from medium. Rates for the component processes (transport of NH₄⁺ across the membrane = R_v assimilation of tissure NH₄⁺ into soluble N compounds = R_a, assimilation of tissue NH₄⁺ into soluble N compounds = R_a and incorporation of soluble N compounds into macromolecules = R₁) were determined by measuring the rate of labelling of the major tissue N pools after the addition of ¹⁵N-ammonium. The results indicate that nitrogen-specific rates (mass N taken up / mass N present / unit time) are ranked in the order of R_t < R_a < R₁ Absolute uptake rates (μmol N. mg dry wt^?1. h^?1) showed a different relationship. Membrane transport appears to be inhibited when NH₄⁺ accumulates in the tissue. Maximum uptake rates occur when assimilation of NH₄⁺ into soluble N compounds begins. Assimilation of NH₄⁺ into soluble N compounds was initially faster than incorporation of soluble N compounds into macromolecules. Implications of rate limitations caused by differences in maximal rates and maximal pool sizes are discussed.     

SEXUAL REPRODUCTIO IN CODIUM FRAGILE SSP. TOMENTOSOIDES (CHLOROPHYCEAE) FROM THE NORTHEAST COAST OF NORTH AMERICA1

Male and female gametes were formed within the same gametangiurn on reproductive plants of Codium fragile (Sur.) Hariot ssp. tomentosoides (Van Goor) Silva growing off Appledore Island in the Gulf of Maine, USA. The female gametes were approximately twice the diameter of male gametes and outnumbered male gametes in the same gametangium by approximately six times. Fusion appears to require gametes from different gametangia if not different plants.     

OLISTHODISCUS LUTEUS (CHRYSOPHYCEAE). III. UPTAKE AND UTILIZATION OF NITROGEN AND PHOSPHORUS1

Uptake and assimilation of nitrogen and phosphorus were studied in Olisthodiscus luteus Carter. A diel periodicity in nitrate reductase activity was observed in log and stationary phase cultures; there was a 10-fold difference in magnitude between maximum and minimum rates, but other cellular features such as chlorophyll a, carbon, nitrogen, C:N ratio (atoms) · cell^?1 were less variable. K_s values (~2 μM) for uptake of nitrate-N and ammonium-N were observed. Phosphorus assimilated · cell^?1· day^?1 varied with declining external phosphorus concentrations; growth rates <0.5 divisions · day^?1 were common at <0.5 μM PO_4-P. Phosphate uptake rates (K_s= 1.0–1.98 μM) varied with culture age and showed multiphasic kinetic features. Alkaline phosphatase activity was not detected. Comparisons of the nutrient dynamics of O. luteus to other phytoplankton species and the ecological implications as related to the phytoplankton community of Narragansett Bay (Rhode Island) are discussed.     

EFFECT OF MALTOSE RELEASE ON UPTAKE AND ASSIMILATION OF AMMONIUM BY SYMBIOTIC CHLORELLA (CHLOROPHYTA)1

When incubated at pH 4–5, Chlorella freshly isolated from symbiosis with Hydra viridissima PALLAS 1766 (green hydra) release large amounts of photosynthetically fixed carbon in the form of maltose, and assimilation of inorganic N is inhibited. Physiological responses to N starvation of the cultured 3N813A strain of maltose-releasing Chlorella differed from those caused by 48 h of maltose release induced by low pH. N starvation increased rates of ammonium assimilation at pH 7.0 in light or darkness, and ammonium assimilation in darkness stimulated cell respiration. In contrast, cells pretreated at pH 5.0 to induce maltose release were unable to take up ammonium at pH 7.0 unless supplied with an external carbon source such as bicarbonate, acetate, or succinate, and rates of uptake were similar to control cells. Freshly isolated symbionts displayed a similar dependency. Rates of ammonium uptake by cells pretreated at pH 5.0 were reduced in darkness and did not stimulate cell respiration. N-starved cells supplied with ammonium also showed a large short-term increase in glutamine pools at the expense of glutamate, as might be expected if large amounts of ammonium were rapidly assimilated via glutamine synthetase/glutamate synthase, whereas after long-term maltose release cells showed only a small increase in glutamine when supplied with ammonium. Furthermore, maltose release caused a fall in pool sizes of a number of amino acids, including glutamine and glutamate, and also caused a decrease in pool sizes of 2-oxoglutarate and phospho-enol-pyruvate, which are required for ammonium assimilation into amino acids. Cells stimulated to synthesize and release maltose may be unable to assimilate ammonium and synthesize amino acids because of diversion of fixed carbon from N metabolism. We estimate that 40–50% affixed C is required for maximal maltose synthesis, whereas up to 30% fixed C is required for ammonium assimilation. These results are discussed in the context of host regulation of symbiotic algal growth.     

EFFECTS OF TEMPERATURE,NITROGEN SUPPLY,AND TISSUE NITROGEN ON AMMONIUM UPTAKE RATES OF THE CHLOROPHYTE SEAWEEDS ULVA CURVATA AND CODIUM DECORTICATUM1

Nitrogen uptake rates of Ulva curvata (Kütz.) de Toni (Ulvales) and Codium decorticatum (Woodw.) Howe (Caulerpales) grown under several N addition regimes were determined by perturbation and continuous mode techniques, and as N demand, by the product of growth rate and tissue N. Uptake rates are reported as the slope of rate vs. concentration curves in each case. N uptake rates of U. curvata were inversely correlated with tissue N and affected only slightly by temperature. There was no correlation of N uptake rate with tissue N in C. decorticatum. N uptake rates of C. decorticatum were affected by temperature but to a lesser degree than were growth rates. Neither N addition per se nor light affected N uptake capacity of either species. The proximal mechanism for seaweeds accumulation of N at low light and temperatures may be that N uptake is less limited by light and temperature than is growth. This in turn may partially compensate for the effects of reduced light and temperature on growth by increasing pigment and enzyme levels. Perturbation uptake rates were higher than continuous mode or N demand rates in Ulva but not in Codium. N uptake rates of Ulva were higher than those of Codium, but N storage capacities were lower. These two observations suggest that Ulva experiences a fundamentally more variable N supply than does Codium. This is consistent with the clarification of Ulva as an ephemeral form and of Codium as persistent. A seaweed's functional form therefore appears to influence the spectrum of resource variability available to it as well as its ability to persist in the environment.     

MAXIMUM AMMONIUM UPTAKE RATES OF SCENEDESMUS QUADRICAUDA (CHLOROPHYTA) AND MICROCYSTIS NOVACEKII (CYANOBACTERIA) GROWN UNDER NITROGEN LIMITATION AND IMPLICATIONS FOR COMPETITION1

We measured maximum ammonium uptake rates of the green alga Scenedesmus quadricauda (Turpin) Brébisson and the blue-green alga Microcystis novacekii (Kom.) Comp. grown in nitrogen (ammonium)–limited chemostats. Maximum uptake rates per cellular carbon were larger in S. quadricauda than in M. novacekii. These rates increased with increased specific growth rates. Maximum uptake rates per cellular nitrogen were also larger in S. quadricauda than in M. novacekii. The maximum uptake rates per cellular nitrogen were nearly constant against increased cellular N:C ratios under nitrogen-limited conditions. The higher maximum uptake rates indicate that S. quadricauda had higher uptake abilities for ammonium than M. novacekii when grown under nitrogen limitation. We examined the competition between both species under two distinct nutrient supply modes, using measured maximum uptake values and computer simulations. Microcystis novacekii prevailed in the small-pulse, high-frequency nutrient supply mode, whereas S. quadricauda became competitively superior in the large-pulse, low-frequency nutrient supply mode. These results indicate that we could control nuisance blooms of blue-green algae in lakes and reservoirs by changing the nutrient supply modes.     

THE RELATIVE IMPORTANCE OF WATER MOTION ON NITROGEN UPTAKE BY THE SUBTIDAL MACROALGA ADAMSIELLA CHAUVINII (RHODOPHYTA) IN WINTER AND SUMMER1

The influence of seawater velocity (1.5–12 cm · s^?1) on inorganic nitrogen (N) uptake by the soft‐sediment perennial macroalga Adamsiella chauvinii (Harv.) L. E. Phillips et W. A. Nelson (Rhodophyta) was determined seasonally by measuring uptake rate in a laboratory flume. Regardless of N tissue content, water velocity had no influence on NO₃^? uptake in either winter or summer, indicating that NO₃^?‐uptake rate was biologically limited. However, when thalli were N limited, increasing water velocity increased NH₄⁺ uptake, suggesting that mass‐transfer limitation of NH₄⁺ is likely during summer for natural populations. Uptake kinetics (V_max, K_s) were similar among three populations of A. chauvinii at sites with different mean flow speeds; however, uptake rates of NO₃^? and NH₄⁺ were lower in summer (when N status was generally low) than in winter. Our results highlight how N uptake can be affected by seasonal changes in the physiology of a macroalga and that further investigation of N uptake of different macroalgae (red, brown, and green) during different seasons is important in determining the relative influence of water velocity on nutrient uptake.     

TRANSIENT AMMONIUM UPTAKE IN THE MACROALGA ULVA LACTUCA (CHLOROPHYTA): NATURE,REGULATION, AND THE CONSEQUENCES FOR CHOICE OF MEASURING TECHNIQUE1

The nature of transient ammonium uptake by the macroalga Ulva lactuca L. was studied from the depletion of ammonium after single additions of ammonium to batch cultures. The experiments were carried out by the application of two different experimental setups: the “multiple flask” and the “perturbation” techniques. Uptake rate was nonlinear with time, and three distinct, succeeding phases of uptake were identified: 1) “surge” uptake, i.e. transiently enhanced uptake that lasted for a few hours only, followed by 2) “internally” controlled uptake, a relatively constant phase occurring at high substrate concentrations, and finally 3) the “externally” controlled uptake phase, which was substrate-dependent and occurred at low substrate concentrations. Surge uptake occurred over a broad range of substrate concentrations but was concentration-dependent and, so, equalled externally controlled uptake rates at substrate concentrations below 3–10 μM. The transient nature of ammonium uptake rate seemed related to rapid changes in small intracellular pools of inorganic nitrogen or amino acids rather than to changes in total N content of the algae. The transient nature of ammonium uptake has important implications for the measurement of uptake rates when either of the two standard methods, the multiple flask and the perturbation technique, are used, and I recommend that a combination of the two methods be used for future uptake experiments.     

ULTRAVIOLET-B RADIATION EFFECTS ON INORGANIC NITROGEN UPTAKE BY NATURAL ASSEMBLAGES OF OCEANIC PLANKTON1

Ultraviolet-B radiation (UVBR: 290-320 nm) inhibited ammonium uptake (p_NH4) and nitrate uptake (p_NO3) in natural plankton assemblages collected during a transect from 37° N to 55° N in the Pacific Ocean. Comparison of responses in p_NH4 to ambient solar- and lamp-enhanced UVBR spectra allowed calculation of an action spectrum for p_NH4 inhibition. The slope of the action spectrum for Pnh₄ is half as steep as action spectra for UVBR inhibition of photosynthetic carbon uptake. Consequently, WBR-induced photoinhibition of p_NH4 extends to greater depths than inhibition of carbon fixation due to the greater relative effect of longer UVBR wavelengths. Inhibition of p_NH4 was dependent upon UVBR dose when doses were weighted by the p_NH4 action spectrum. Dependence of WBR inhibition of p_NH4 on dose rate was not apparent. We found that near-surface p_NH4 and p_NO3 can be overestimated in excess of 50% when measured using standard incubation vessels made of UVBR-absorbing materials such as polycarbonate.     

EFFECT OF NITROGEN SOURCE ON PLATYMONAS (CHLOROPHYTA) CELL COMPOSITION AND AMINO ACID UPTAKE RATES1

The effect of nitrogen source (nitrate, ammonia and/or amino acids) on cell composition and amino acid uptake rates was examined. Substantial levels of free amino acids accumulated intracellularly with all nitrogen sources used. Ammonia accumulated only when provided in the medium. The presence of ammonia in the medium decreased the intracellular accumulation of free amino acids, especially arginine. Amino acid uptake rates were suppressed by the presence of excess nitrogen, especially ammonia. However, the suppression of uptake did not show any particular relation to the nitrogenous cell composition.     

MICROSENSOR STUDIES OF OXYGEN AND LIGHT DISTRIBUTION IN THE GREEN MACROALGA CODIUM FRAGILE1

Scalar irradiance, oxygen concentration, and oxygenic photosynthesis were measured at 0.1 mm spatial resolution within the tissue of the siphonous green macroalga Codium fragile subsp. tomentosoides (van Goor) Silva by fiber-optic scalar irradiance microsensors and oxygen microelectrodes. The scalar irradiance of visible light was strongly attenuated in the outer 0.2 mm of the tissue but was nearly constant for the subsequent 1.0 mm of photo-synthetic tissue. Far-red scalar irradiance at 750 nm increased below the tissue surface to a maximum of 200% of incident irradiance at 1.2 mm depth due to multiple scattering in the medullary tissue. The constant intensity of visible light below 0.2 mm was thus a result of the combined effects of absorption and backscattering from the medulla. The oxygen exchange between the alga and the surrounding water was diffusion-limited with a steep O₂gradient inside and around the alga. In darkness, the tissue below 0.6 mm became anoxic, and endophytic extracellular space provided an environment where anoxygenic microbial processes may occur. When illuminated at 160 nmol photons·^?2·^?1, O₂ concentrations exceeded ambient levels throughout the thallus, with a maximum of 250% of air saturation just below the surface. The amplitude of oxygen variation was buffered by gas bubbles formed in the medullary tissue.     

INORGANIC NITROGEN AND PHOSPHORUS UPTAKE KINETICS IN PALMARIA PALMATA (RHODOPHYTA)1

The N and P uptake responses were studied in a northern Spanish population of the edible red seaweed Palmaria palmata (Linnaeus) Kuntze. The fronds were incubated at different concentrations, and the nutrient depletion in the medium was measured at successive times to calculate uptake rates. Palmaria palmata uptake response was biphasic and nonsaturable for inorganic P. This would allow the species to exploit transient pulses of high P concentration in natural and fertilized conditions. Such a response is a common feature of algae avoiding nutrient deficiency. At average concentrations measured in the ocean, the response was nonsaturable for inorganic N sources, except for ammonium in autumn and winter when it is not the major N source. In contrast to the general rule of ammonium being taken at a higher rate than nitrate, we found similar affinity for both nutrients corresponding to the minor role of ammonium as N source for field populations over the year.     

UPTAKE OF INORGANIC CARBON BY CLADOPHORA GLOMERATA (CHLOROPHYTA) FROM THE BALTIC SEA1

Carbon uptake in the green macroalga Cladophora glomerata (L.) Kütz. from the brackish Baltic Sea was studied by recording changes in pH, alkalinity, and inorganic carbon concentration of the seawater medium during photosynthesis. The use of specific inhibitors identified three uptake mechanisms: 1) dehydration of HCO₃ ^? into CO₂ by periplasmic carbonic anhydrase, followed by diffusion of CO₂ into the cell; 2) direct uptake of HCO₃ ^? via a 4,4′‐diisothiocyanato‐stilbene‐2,2′‐disulfonate‐sensitive mechanism; and 3) uptake of inorganic carbon by the involvement of a vanadate‐sensitive P‐type H ⁺ ‐ATPase (proton pump). A decrease in the alkalinity of the seawater medium during carbon uptake, except when treated with vanadate, indicated a net uptake of the ionic species contributing to alkalinity (i.e. HCO₃ ^? , CO₃^{2 ?} , and OH ^? ) from the medium, where OH ^? influx is equivalent to H ⁺ efflux. This would suggest that the proton pump is involved in HCO₃ ^? transport. We also show that the proton pump can be induced by carbon limitation. The inducibility of carbon uptake in C. glomerata may partly explain why this species is so successful in the upper littoral zone of the Baltic Sea. Usually, carbon limitation is not a problem in the upper littoral of the sea. However, it may occur frequently within dense Cladophora belts with high photosynthetic rates that create high pH and low carbon concentrations in the alga's microenvironment.