Daily timing of emersion and elevated atmospheric CO2 concentration affect photosynthetic performance of the intertidal macroalga Ulva lactuca (Chlorophyta) in sunlight

The lunar day differs in length from the solar day so that times of low tide vary from day to day. Thus, aerial exposure of intertidal seaweeds may be during the day or during the night. We measured photosynthetic CO, assimilation rates of the intertidal green macroalga Ulva lactuca during exposures of varied daily timings during sunny days of summer to establish how photosynthetic performance responds to emersion timing under varied CO2 levels [at ambient (360 ppmv) and 2x ambient (720 ppmv) atmospheric CO2 concentrations]. There was an increase in net photosynthetic rates following some duration of exposure when the initial timing of exposure occurred during early morning (06.30 h) and late afternoon (17.15 h). In contrast, net rates exhibited a sharp decline with exposure duration when the initial timing of exposure occurred at 09.30 h, 15.30 h and especially at noon (12.30 h), implying the occurrence of a severe photoinhibition resulting from mid-day insolation. Doubled atmospheric CO2 concentration significantly enhanced the emersed photosynthetic rates, indicating that the emersed photosynthesis is CO2-limited at ambient CO2 levels. However, increasing CO2 barely stimulates the emersed photosynthetic rates during mid-day insolation.

Responses of dark respiration in the light to desiccation and temperature in the intertidal macroalga, Ulva lactuca (Chorophyta) during emersion

Dark respiration (nonphotorespiratory mitochondrial CO2 release) in the light (R-L) of the intertidal macroalga Ulva lactuca (Chorophyta) during emersion was investigated with respect to its response to variations in temperature and desiccation. R-L was estimated by CO2 gas-exchange analysis using the Kok effect method, whereas dark respiration in darkness (R-D) was determined from CO2 release at zero light. Rates of R, were significantly and consistently lower than those of R-D in emersed U. lactuca across all the temperature and desiccation levels measured. This demonstrated that dark respiration was partially depressed in the light, with the percentage inhibition ranging from 32 to 62%. Desiccation exerted a negative effect on R-L and R-D at a high temperature, 33 degrees C, whereas it had much less effect on respiration at low and moderate temperatures, 23 and 28 degrees C. In general, R-L and R-D increased with increasing temperature in U. lactuca during all stages of emersion but responded less positively to temperature change with increasing desiccation. Additionally, the Q(10) value (i.e. the proportional increase of respiration for each 10 degrees C rise in temperature) for R-L calculated over the temperature range of 23 to 33 degrees C was significantly higher than that for R-D in U. lactuca during the initial stages of emersion. Respiratory carbon loss as a percentage of gross photosynthetic carbon gain increased with increasing temperature and/or desiccation but was significantly reduced when estimated using R-L rather than R-D. It is suggested that measurements of R-L and how it changes in a variable environment are as important as estimates of R-D and photosynthesis in determining simultaneous balance between photosynthetic carbon uptake and respiratory carbon loss and in modeling the net daily carbon gain for an intertidal macroalga.

Allelopathic interactions between the macroalga Ulva pertusa and eight microalgal species

HETEROSIGMA-AKASHIWO RAPHIDOPHYCEAE; CENTRAL VENICE LAGOON; ALEXANDRIUM-TAMARENSE; RED-TIDE; COASTAL LAGOONS; PHYTOPLANKTON; GROWTH; BAY; DINOFLAGELLATE; COMPETITION

Construction and characterization of a bacterial artificial chromosome library of marine macroalga Porphyra yezoensis (Rhodophyta)

A large-DNA-fragment library is necessary for research into the Porphyra genome. In this study, a bacterial artificial chromosome (BAC) library of Porphyra yezoensis was constructed and characterized. The library contains 54,144 BAC clones with an average insert size of about 65 kb and fewer than 0.7% of clones without large inserts. Therefore, its capacity is more than 6.6 P. yezoensis genome equivalents, and the probability of recovering any nuclear DNA sequence from the library is higher than 99%. The library shows good fidelity and stability. A putative trehalose-6-phosphate synthase (TPS) gene was successfully screened out from the library. The above results show that the library is useful for gene cloning and genomic research in P. yezoensis.

Bioremediation potential of the macroalga Gracilaria lemaneiformis (Rhodophyta) integrated into fed fish culture in coastal waters of north China

Fed fish farms produce large amounts of wastes, including dissolved inorganic nitrogen and phosphorus. In China, fish mariculture in coastal waters has been increasing since the last decade. However, there is no macroalgae commercially cultivated in north China in warm seasons. To exploit fish-farm nutrients as a resource input, and at the same time to reduce the risk of eutrophication, the high-temperature adapted red alga Gracilaria lemaneiformis (Bory) Dawson from south China was co-cultured with the fish Sebastodes fuscescens in north China in warm seasons. Growth and nutrient removal from fish culture water were investigated in laboratory conditions in order to evaluate the nutrient bioremediation capability of G. lemaneiformis. Feasibility of integrating the seaweed cultivation with the fed fish-cage aquaculture in coastal waters of north China was also investigated in field conditions. Laboratory seaweed/fish co-culture experiments showed that the seaweed was an efficient nutrient pump and could remove most nutrients from the system. Field cultivation trials showed that G. lemaneiformis grew very well in fish farming areas, at maximum growth rate of 11.03% day(-1). Mean C, N, and P contents in dry thalli cultured in Jiaozhou Bay were 28.9 +/- 1.1%, 4.17 +/- 0.11 % and 0.33 +/- 0.01 %, respectively. Mean N and P uptake rates of the thalli were estimated at 10.64 and 0.38 mu mol g(-1) dry weight h(-1), respectively. An extrapolation of the results showed that a 1-ha cultivation of the seaweed in coastal fish fanning waters would give an annual harvest of more than 70 t of fresh G. lemaneiformis, or 9 t dry materials; 2.5 t C would be produced, and simultaneously 0.22 t N and 0.03t P would be sequestered from the seawater by the seaweed. Results indicated that the seaweed is suitable as a good candidate for seaweed/fish integrated mariculture for bioremediation and economic diversification. The integration can benefit economy and environment in a sustainable manner in warm seasons in coastal waters of north China. (c) 2005 Elsevier B.V. All rights reserved.

Photosynthetic utilization of inorganic carbon in the economic brown alga, Hizikia Fusiforme (Sargassaceae) from the South China Sea

The mechanism of inorganic carbon (C-i) acquisition by the economic brown macroalga, Hizikia fusiforme (Harv.) Okamura (Sargassaceae), was investigated to characterize its photosynthetic physiology. Both intracellular and extracellular carbonic anhydrase (CA) were detected, with the external CA activity accounting for about 5% of the total. Hizikia fusiforme showed higher rates of photosynthetic oxygen evolution at alkaline pH than those theoretically derived from the rates of uncatalyzed CO2 production from bicarbonate and exhibited a high pH compensation point (pH 9.66). The external CA inhibitor, acetazolamide, significantly depressed the photosynthetic oxygen evolution, whereas the anion-exchanger inhibitor 4,4'-diisothiocyano-stilbene-2,2'-disulfonate had no inhibitory effect on it, implying the alga was capable of using HCO3- as a source of C-i for its photosynthesis via the mediation of the external CA. CO2 concentrations in the culture media affected its photosynthetic properties. A high level of CO2 (10,000 ppmv) resulted in a decrease in the external CA activity; however, a low CO2 level (20 ppmv) led to no changes in the external CA activity but raised the intracellular CA activity. Parallel to the reduction in the external CA activity at the high CO2 was a reduction in the photosynthetic CO2 affinity. Decreased activity of the external CA in the high CO2 grown samples led to reduced sensitiveness of photosynthesis to the addition of acetazolamide at alkaline pH. It was clearly indicated that H. fusiforme, which showed CO2-limited photosynthesis with the half-saturating concentration of C-i exceeding that of seawater, did not operate active HCO3- uptake but used it via the extracellular CA for its photosynthetic carbon fixation.

Photosynthetic responses to inorganic carbon in Ulva lactuca under aquatic and aerial states

Intertidal macroalgae experience continual alternation of photosynthesis between aquatic state at high tide and aerial state at low tide. The comparative photosynthetic responses to inorganic carbon were investigated in the common intertidal macroalga Ulva lactuca L. along the coast of Shantou between aquatic and aerial state. The inorganic carbon dissolved in seawater at present could fully (at 10 degreesC or 20 degreesC) or nearly (at 30 degreesC) saturate the aquatic photosynthesis of U. lactuca. However, the aerial photosynthesis was limited by current ambient atmospheric CO2 level, and such a limitation was more severe at higher temperature (20degrees - 30degrees T) than at lower temperature (10 T). The carbon-saturated maximal photosynthesis of U. lactuca under aerial state was much greater than that under aquatic state at 10 degreesC and 20 degreesC, while the maximal photosynthesis under both states was similar at 30 degreesC. The aerial values of K-m (CO2) for photosynthesis were higher than the aquatic values. On the contrary, the values of apparent photosynthetic CO2 conductance under aerial state were considerably lower than that under aquatic state. It was concluded that the increase of atmospheric CO2 would enhance the primary productivity of U. lactuca through stimulating the photosynthesis under aerial state during low tide.

Photosynthetic parameters of sexually different parts of Porphyra katadai var. hemiphylla (Bangiales, Rhodophyta) during dehydration and re-hydration

Physiological data from extreme habitat organisms during stresses are vital information for comprehending their survival. The intertidal seaweeds are exposed to a combination of environmental stresses, the most influential one being regular dehydration and re-hydration. Porphyra katadai var. hemiphylla is a unique intertidal macroalga species with two longitudinally separated, color distinct, sexually different parts. In this study, the photosynthetic performance of both PSI and PSII of the two sexually different parts of P. katadai thalli during dehydration and re-hydration was investigated. Under low-grade dehydration the variation of photosystems of male and female parts of P. katadai were similar. However, after the absolute water content reached 42%, the PSI of the female parts was nearly shut down while that of the male parts still coordinated well and worked properly with PSII. Furthermore, after re-hydration with a better conditioned PSI, the dehydrated male parts were able to restore photosynthesis within 1 h, while the female parts did not. It is concluded that in P. katadai the susceptibility of photosynthesis to dehydration depends on the accommodative ability of PSI. The relatively lower content of phycobiliprotein in male parts may be the cause for a stronger PSI after severe dehydration.

Suspension culture of gametophytes of transgenic kelp in a photobioreactor

Transgenic Laminaria japonica gametophytes producing a recombinant tissue-type plasminogen activator (rtPA) protein, which is an effective third-generation thrombolytic agent for acute myocardial infarction (AMI), were cultured in an illuminated bubble column bioreactor. A maximum final dry cell weight of 1120 mg l(-1) was obtained in batch culture with an initial dry cell weight of 126 mg l(-1) and with aeration rate of 1.2 l air min(-1) l(-1) culture, nitrate at 1.5 mM and phosphate at 0.17 mM. The yield of rtPA was 56 mu g g(-1) dry cell wt. This is the first report regarding cultivation of a transgenic macroalga in a bioreactor.

Cultivation of the brown alga Hizikia fusiformis (Harvey) Okamura: controlled fertilization and early development of seedlings in raceway tanks in ambient light and temperature

Commercial cultivation of the dioecious brown macroalga Hizikia fusiformis (Harvey) Okamura in East Asia depends on the supply of young seedlings from regenerated holdfasts or from wild population. Recent development of synchronized release of male and female gametes in tumble culture provides a possibility of mass production of young seedlings via sexual reproduction. In this paper, we demonstrate that controlled fertilization can be efficiently realized in ambient light and temperature in a specially designed raceway tank in which the sperm-containing water has been recirculated. The effective fertilization time of eggs by sperm was found to be within six hours. Fast growth and development of the young seedlings relied on the presence of water currents. Velocity tests demonstrated that young seedlings of 2-3 mm in length could withstand a water current of 190 cm s(-1) stop without detachment. Culture experiments at 24 h postfertilization showed that elongation of both the seedlings and their rhizoids were not hampered by high irradiance up to 600 mu mol photons m(-2) stop s(-1) stop. However, growth was slightly retarded if cultured at a temperature of 16 degrees C compared to other culture temperatures of 22, 25 and 29 degrees C. No seedling detachment was observed after transfer of the young seedlings to raft cultivation in the sea after one and 1.5 months post-fertilization, indicating the feasibility of obtaining large quantity of seedlings in such a system.

Optimization of conditions for cell cultivation of Porphyra haitanensis conchocelis in a bubble-column bioreactor

Process conditions for cell cultures derived from conchocelis of female red macroalga Porphyra haitanensis were optimized in an illuminated 0.3-l bubble-column photobioreactor, using CO2 in air as the sole carbon source during a 20-day cultivation period. It reached the highest growth rate when the initial cell density was 700 mg l(-1)(dry weight), the optional aeration rate was 1.2 v/v/min, inorganic nitrate concentration was 15 mM and inorganic phosphate concentration was 0.6 mM. This is the first reported bioreactor cultivation study of cell cultures derived from conchocelis of Porphyra haitanensis.

Effects of macroalgae Ulva pertusa (Chlorophyta) and Gracilaria lemaneiformis (Rhodophyta) on growth of four species of bloom-forming dinoflagellates

The effects of fresh thalli and culture medium filtrates from two species of marine macroalgae, Ulva pertusa Kjellm (Chlorophyta) and Gracilaria lemaneiformis (Bory) Dawson (Rhodophyta), on growth of marine microalgae were investigated in co-culture under controlled laboratory conditions. A selection of microalgal species were used, all, being identified as bloom-forming dinoflagellates: Prorocentrum donghaiense Lu sp., Alexandrium tamarense (Lebour) Balech, Amphidinium carterae Hulburt and Scrippsiella trochoide (Stein) Loeblich III. Results showed that the fresh thalli of either U. pertusa or G. lemaneiformis significantly inhibited the microalgal growth, or caused mortality at the end of the experiment. However, the overall effects of the macroalgal culture filtrates on the growth of the dinoflagellates were species-specific (inhibitory, stimulatory or none) for different microalgal species. Results indicated an allelopathic effect of macroalga on the co-cultured dinoflagellate. We then took P. donghaiense as an example to further assess this hypothesis. The present study was carried out under controlled conditions, thereby excluded the fluctuation in light and temperature. Nutrient assays showed that nitrate and phosphate were almost exhausted in G. lemaneiformis co-culture. but remained at enough high levels in U pertusa co-culture, which were well above the nutrient limitation for the microalgal growth, when all cells of P. donghaiense were killed in the co-culture. Daily f/2 medium enrichment greatly alleviated the growth inhibition on P. donghaiense in G. lemaneiformis co-culture, but could not eliminate it. Other environmental factors, such as carbonate limitation, bacterial presence and the change of pH were also not necessary for the results. We thus concluded that allelopathy was the most possible reason leading to the negative effect of U. pertusa on P. donghaiense, and the combined roles of allelopathy and nutrient competition were essential for the effect of G. lemaneiformis on P. donghaiense. (c) 2006 Elsevier B.V. All rights reserved.

Food sources of three bivalves living in two habitats of Jiaozhou Bay (Qingdao, China): Indicated by lipid biomarkers and stable isotope analysis

Food Sources of three filter-feeding bivalves from two habitats (intertidal oyster Crassostrea gigas, mussel Mytilus galloprovincialis. and subtidal cultured scallop Chlamys farreri) of Jiaozhou Bay (Qingdao,China) were determined by fatty acid and stable isotope in analysis. Cultured scallop was characterized by significant diatom markets such as 16:1/16:0 close to 1 and high ratio of 20:5(n - 3)/22:6(n - 3), hence we assume that the scallop mainly feeds on diatoms. Fatty acid biomarkers specific to bacteria and terrestrial materials were also found in considerable amounts in scallop tissue, which suggested that there were Substantial bacterial and terrestrial input into the food of the species. Intertidal oyster and mussel, however, exhibited significant flagellate marker. 22:6(n - 3). and lower level of diatom markers. which indicated that flagellates are also part of intertidal bivalves' Planktonic food Sources: meanwhile, high level of Chlorophyta fatty acid marker, Sigma 18:2(n - 6) + 18:3(n - 3), suggested that Ulva pertusa (Chlorophyta) seaweed bed supplied important food sources to intertidal bivalves. Additionally, result of stable isotope analysis showed that phytoplankton contributed 86.2 to 89.0% to intertidal bivalves' carbon budget; macroalga U. pertusa origin source had a contribution of MIX, to 11.0%, which indicated its role Lis in important supplemental food source to intertidal bivalves. From this study. it is concluded that the dietary difference of three bivalves probably relates to the different potential food sources in the scallop farm and intertidal zone in Jiaozhou Bay.