Studies on R-phycoerythrins from two Antarctic marine red algae and a mesophilic red alga

R-phycoerythrin was purified from two benthic red algae, Iridaea cordata and Phyllophora antarctica, obtained growing at ?2°C under thick sea ice off the coast of Antarctica. For the I. cordata protein, the molecular mass was 245,000 Da, and its secondary structure was 60% α helix, 17% β sheet, 16% turn, and 7% other. The light-harvesting faculties of the I. cordata protein resembled those of R-phycoerythrins from mesophilic red algae and were distinctive from the novel R-phycoerythrin from P. antarctica. Deconvolution of the visible absorption spectrum of R-phycoerythrin from I. cordata indicated a minimum of five component bands having maxima at 568, 558, 534, 496, and 481?nm. R-phycoerythrins from the mesophilic Porphyra tenera and psychrophilic Phyllophora antarctica had the same five bands. The protein from Phyllophora antarctica obtained its unique spectrum from a more intense component at 482?nm, and a less intense band at 533?nm. This change was probably produced by a replacement of phycoerythrobilin by phycourobilin. A temperature study of the circular dichroism CD was obtained for R-phycoerythrin from I. cordata from 4 to 80°C. Laser time-resolved fluorescence studies on R-phycoerythrin showed bilin to bilin energy transfer with a 60.2-ps lifetime, which should occur by the Förster resonance. The similarities in spectra between the proteins from I. cordata and Porphyra tenera and the different spectrum for the protein from Phyllophora antarctica show that only particular antarctic habitats require unique R-phycoerythrins.     

Macroalgal photosynthesis near the southern global limit for growth; Cape Evans,Ross Sea,Antarctica

Photosynthetic characteristics of the red macroalgae Phyllophora antarctica and Phymatolithon foecundum collected from under sea ice at Cape Evans, McMurdo Sound (Ross Sea) were determined using in situ fluorometric and lab-based oxygen exchange techniques. Only 0.16% of incident irradiance penetrated the 2.5 m thick ice cover and photosynthetic parameters for both taxa were characteristic of highly shade-adapted plants. Saturation onset parameter (E _k) did not exceed 13 mol photons m^-2 s^-1 in either taxon. For Phyllophora antarctica the light saturated photosynthetic rate at –1^°C was 10 mol O₂ g^-1 FW h^-1 and respiration averaged 3.3 mol O₂ g^-1 FW h^-1 between sampled depths of 10 and 25 m. A light meter deployed at 15 m depth for a year recorded a marked increase in underwater irradiance on the last day of January 2002 coinciding with ice-breakout, and a maximum value for irradiance of 120 mol photons m^-2 s^-1 on 9 February 2002. The 2-month ice-free period was the only time when irradiance consistently exceeded compensation (photosynthesis=respiration) and enabled Phyllophora antarctica to accumulate sufficient carbon to result in a measurable increase in thallus area equivalent to a biomass increment of 1.87 mg (DW) per frond. Near the southern global limit for marine macroalgae, conditions that dictate the availability of underwater irradiance are extremely variable from year to year. Low respiration rates enhance longevity of the Phyllophora antarctica thallus, enabling it to not only survive the winter darkness, but also to retain photosynthetic capacity and thus take advantage of windows of higher irradiance.     

Ion-exchange properties of cell walls of red seaweed Phyllophora crispa

Research into ion-exchange properties of cell walls isolated from thallus of red seaweed Phyllophora crispa was carried out. Ion-exchange capacity and the swelling coefficient of the red alga cell walls were estimated at various pH values (from 2 to 12) and at constant ionic strength of a solution (10 mM). It was established that behavior of cell walls as ion-exchangers is caused by the presence in their matrix of two types of cation-exchange groups and amino groups. The amount of the functional group of each type was estimated, and the corresponding values of pK(a) were calculated. It can be assumed that ionogenic groups with pK(a) -5 are carboxyl groups of uronic acids, and ionogenic groups with pK(a) -7.5 are carboxyl groups of the proteins. Intervals of pH in which cation-exchange groups are ionized and can take part in exchange reactions with cations in the environment are defined. It was found that protein was a major component of cell wall polymeric matrix because its content was 36%.     

Systematic studies of the antarctic species of the phyllophoraceae (Gigartinales,Rhodophyta) based on rbcL sequence analysis

The taxonomic placement of four antarctic species of the marine red algal family Phyllophoraceae (Gigartinales) is assessed within a preliminary molecular phylogeny of the family based on direct sequence analysis of the chloroplast gene rbcL. Parsimony analysis of rbcL sequences indicates that Gymnogongrus antarcticus and Gymnogongrus turquetii cluster in a clade consisting predominantly of southern hemisphere species currently placed in Gymnogongrus and Ahnfeltiopsis, whereas Phyllophora ahnfeltioides and Phyllophora antarctica cluster in a separate clade that is widely divergent from the northern hemisphere Phyllophora clade. Results from molecular and morphological data challenge the current taxonomic concept that type of life history is a phylogenetically valid criterion for recognition of genera in the Phyllophoraceae.     

Animal associations with the dominant species of shallow water macrophytes along the coastline of the Vestfold Hills,Antarctica

Benthic grab samples, taken through core holes in sea-ice 1983, returned eight species of marine macrophytes which had associated fauna on at least one sample. A total of 24 animal taxa were found on these eight macrophytes. Phyllophora antarctica had a significantly greater biomass of attached fauna per unit of macrophyte biomass than any of the other seven species. It also had the greatest variety of epiphytic fauna (21 taxa). Phyllophora antarctica was the only macrophyte collected from comparatively protected Ellis Fjord, apart from filamentous algae, including Cladophora subsimplex and Geminocarpus austrogeorgiae, which were found in the most landward basin. It therefore played an important role in the ecology of the fjord. Palmaria decipiens was commonly collected at more exposed coastal sites and was dominant in Long Fjord, characterized here as intermediate in terms of water movement. Sixteen animal taxa were collected from this macrophyte with the greatest numbers being found in July and August. Only four animal taxa were collected from Iridaea cordata. The other five macrophyte species from which epiphytic animals were collected (Porphyra endiviifolium, Plocamium cartilagineum, Desmarestia menziesii, Himantothallus grandifolius and Cladophora subsimplex had animals associated with them on less than half the occasions when they were found.     

The deposition and mortality of the eggs of the Baltic herring Clupea harengus membras L., on different substrates in the south-west archipelago of Finland

The environmental factors affecting the deposition and mortality of the eggs of the spring-spawning Baltic herring were studied in the inner Archipelago Sea of south-western Finland. On four spawning grounds, 27 study squares (area 1 m²) were surveyed by divers. In each square, one quantitative egg sample was taken and the following data were recorded: depth, temperature, bottom quality of the square and under the eggs, egg substrate, cover (%) of plants and/or the bivalve Mytilus edulis , and the heterogeneity of the environment, expressed as the total number of all bottom materials, plants and Mytilus found in the square.
Eggs were found in the depth zone of 1–4 m. Their density ranged from 6000 to 2·3 millions of eggs per square metre. Egg number had a significant negative correlation with both depth (r=-0·545, P <0·01, d.f. = 25) and temperature of the sea water ( r =-0.479,. P <0·01,d.f. = 25), and a positive correlation with the total cover (%) in the squares ( r = 0·375, P < 0·05, d.f. = 25).
The highest numbers of eggs were found on Cladophora glomerata, which the most preferred spawning substrate of those present. Mortality of the eggs varied according to the substrate. In eggs attached to Cladophora, Potamogeton and Mytilus, the proportion of dead eggs was 0.15·5%; in those attached to red algae it was significantly higher ( Furcellaria , 5·0–63·2%; Phyllophora , 0–95·2%).     

Photoinhibition of Photosynthesis and its Recovery in Red Algae

In different marine red algae (Chondrus crispus, Delesseria sanguinea, Membranoptera alata, Phycodrys rubens, Phyllophora truncata, Polyneura hilliae) photoinhibition of photosynthesis has been investigated by means of both fluorescence and oxygen measurements. Measurements of absolute oxygen production show that photoinhibition causes a decline in the initial slope and in the rate of bending of the fluence rate-response curve (i.e. the photosynthetic efficiency at non-saturating fluence rates), as well as a decline in the photosynthetic capacity (Pm) at saturating fluence rates. Fluorescence data (Fv/Fm) were consistent with the results of oxygen measurements. Under excessive light photoinhibition protects photosynthesis against photo-damage in red algae. However, an increase in the initial fluorescence (Fo) after photoinhibitory treatment indicates that it could not prevent photodamage entirely. Action spectra of photoinhibition demonstrate that the main photoinhibition site in Polyneura hiliae is PS II, because far red light absorbed by PS I was ineffective. The strong increase of Fo in the blue wavelength range and the slight and partial recovery in weak blue light indicate that blue light especially causes photodamage. Recovery of photosynthesis requires dim white light conditions. Experiments with monochromatic light also show a wavelength dependence of recovery. Moreover, the recovery of photosynthesis after a photoinhibitory treatment is strongly temperature dependent, indicating participation of enzymatic processes. The comparison of fluorescence and oxygen measurement of the recovery shows different results in some species. The rate of oxygen production in red control light increased immediately after photoinhibited algae were exposed to weak light conditions. Surprisingly, the ratio of variable to maximum fluorescence (Fv/Fm) of Phyllophora truncata and the maximum fluorescence (Fm) of Polyneura hilliae show first a delay of the recovery under weak light conditions. Thus, in recovery experiments fluorescence and oxygen data are not quite consistent.     

Variation in distribution and biomass of marine benthic algae off Kulien, S Sweden

Samples of the macrobenthic flora were collected at 2, 4, 6, 8 and 10 m below mean water level at Kullen, S Sweden. The frequency, length, reproductive pattern and biomass of the commonest species, and the variations of these with depth and season are discussed. The species composition and the size of the plants indicate that the flora is strongly affected by the brackish environment. Only few species contributed substantially to the total mean biomass (100.1 g dry wt 0.25 m–2): Fucus Serratus (56%), Laminaria digitata (7%), L. saccharina (4%), Furcellaria lumbricalis (14%), the Phyllophora species (5%) and Chondrus crispus (3%). Species with thin filamentous or sheet–like thalli made up the remaining ca. 11%. The contribution made by this group increased with depth from 1% at 2 m to 48% at 10 m. The mean biomass at each depth was strongly correlated to the irradiance. The seasonal variation in biomass followed that of the irradiance but with a lag. In winter and late summer the growth was also influenced by temperature and nutrient concentrations. The total biomass is delimited by the low and fluctuating salinity in Öresund.     

The distribution of Ceratocolax hartzii

Ceratocolax hartzii Rosenv. (Fam. Phyllophoraceae) has been found to occur only on Phyllophora brodiaei (Turn.) Endlich. and its various forms. New records reported here confirm that it is truly circumboreal and widespread in the Arctic seas, and show that its range in the Atlantic extends south to Rhode Island, U.S.A., and Ireland.     

Ion-exchange properties of cell walls of red seaweed <Emphasis Type="Italic">Phyllophora crispa</Emphasis>

Research into ion-exchange properties of cell walls isolated from thallus of red seaweed Phyllophora crispa was carried out. Ion-exchange capacity and the swelling coefficient of the red alga cell walls were estimated at various pH values (from 2 to 12) and at constant ionic strength of a solution (10 mM). It was established that behavior of cell walls as ion-exchangers is caused by the presence in their matrix of two types of cation-exchange groups and amino groups. The amount of the functional group of each type was estimated, and the corresponding values of pK _a were calculated. It can be assumed that ionogenic groups with pK _a ∼5 are carboxyl groups of uronic acids, and ionogenic groups with pK _a ∼7.5 are carboxyl groups of the proteins. Intervals of pH in which cation-exchange groups are ionized and can take part in exchange reactions with cations in the environment are defined. It was found that protein was a major component of cell wall polymeric matrix because its content was 36%.     

On background levels of heavy metals in agarophytes of the Black Sea

Ecological standards for commercial seaweeds are very important due to anthropogenic pollution of the marine sublittoral. The content of heavy metals in agarophyte red algae, seawater, and sediments are investigated by the method of atomic absorption spectroscopy. The level of Cd, Pb, and Ni in Gracilaria verrucosa was low, while the level of Zn, Pb, and Nu was comparatively higher in Gelidium latifolium and Phyllophora nervosa, especially in the Caucasus nearshore zone. There is no relationship between the concentration of metals in the environment and the concentration of metals in agarophytes. Background levels of dangerous metals are discussed in connection with commercial exploitation of red algae.     

ECOTYPIC VARIATION IN PHYLLOPHORA PSEUDOCERANOIDES (RHODOPHYTA) ENSURES WINTER REPRODUCTION THROUGHOUT ITS GEOGRAPHIC RANGE1

Responses to temperature and daylength were determined in laboratory culture for isolates of the red alga Phyllophora pseudoceranoides (Gmelin) Newroth et A.R.A. Taylor from Nova Scotia, Iceland, Roscoff (France), and Helgoland (Germany). All isolates grew from 3° to 25° C and survived from -2° or 0° C to 27° C but not 30° C. Reproductive requirements differed between life history phases and isolates. Isolates from Helgoland and Roscoff formed sporangial sori at 3°-20° C, tetraspores at 3°-12° C, and procarps at 10°-20° C, irrespective of daylength. Spermatangia developed at 10°-23° C but only in long days. As the other European isolates, the isolate from Iceland formed tetrasporangia at 3°-12° C, but it had an additional requirement for short days. The Nova Scotian isolate formed sori at 10°-20° C and sporulated at 10°-18° C. When grown plants were transferred from noninductive to inductive conditions, sori were formed after 4 months and tetraspores developed and were shed (1-)3 months later. Procarps formed 1(-3) months after transfer. The phenology of P. pseudoceranoides was studied at Helgoland and Roscoff, where similar seasonal patterns were observed. Plants were perennial, forming new blades from October to June, which degenerated between August and February. In June, reproductive structures (sori, spermatangia, and procarps) started to appear on the new blades. From October to April, mature cystocarps were found. Mature tetrasporangia were observed only in February. The life history of P. pseudoceranoides is regulated by temperature and daylength. Differential effects on the different life history phases all serve to confine the production of spores (both carpospores and tetraspores) to the winter season. Differences in response between isolates from different geographic regions bring about the same effect: spores are shed only in winter. The nature of the geographic boundaries of P. pseudoceranoides is discussed.     

Benthic marine algal flora of the north shore of Prince Edward Island (Gulf of St. Lawrence), Canada

A seaweed community along 60 km of shoreline in the southern Gulf of St. Lawrence has been studied for 5 years. Because of severe ice action along the shores, the algal flora is largely restricted to subtidal levels, which are accessible only during the ice-free months, April to early December. A total of 52 Rhodophyta, 50 Phaeophyta, 19 Chlorophyta and 5 Cyanophyta was identified, mostly occurring between mean low water and 10 m depth. The normally intertidal dominants Focus vesiculosus L. and Chondrus crispus Stackh. flourish at depths of 0–2.5 m and 2.5–5 m, respectively. Chondrus and Furcellaria lumbricalis (Huds.) Lamour. co-dominate the algal benthos at 5–7.5 m, and F. lumbricalis becomes prevalent at 7.5–10 m. Phyllophora truncata (Pallas) A. Zin. and Phyllophora pseudoceranoides (Gmel.) Newr. & A. Tayl. are the main perennials at 10–15 m, and often at 15–20 m as well. Below 15 m, the flora diminishes markedly, and at 20–25 m consists chiefly of Agarum cribrosum (Mert.) Bory, crustose corallines and Polysiphonia urceolata (Lightf. ex Dillw.) Grev. Laminarians other than Chorda occur only as scattered plants or small patches and do not exceed 1 m length. The algal community is relatively undiversified and is especially deficient in green algae. Despite higher surface-water temperatures during summer, the flora is essentially a cold-water one as is indicated by both composition and low R/P and (R+C)/P ratios of 1.06 and 1.45, respectively.     

Abundance and composition of epiphytic bacterial and archaeal ammonia oxidizers of marine red and brown macroalgae

Ammonia-oxidizing bacteria (AOB) and archaea (AOA) are important for nitrogen cycling in marine ecosystems. Little is known about the diversity and abundance of these organisms on the surface of marine macroalgae, despite the algae's potential importance to create surfaces and local oxygen-rich environments supporting ammonia oxidation at depths with low dissolved oxygen levels. We determined the abundance and composition of the epiphytic bacterial and archaeal ammonia-oxidizing communities on three species of macroalgae, Osmundaria volubilis, Phyllophora crispa, and Laminaria rodriguezii, from the Balearic Islands (western Mediterranean Sea). Quantitative PCR of bacterial and archaeal 16S rRNA and amoA genes was performed. In contrast to what has been shown for most other marine environments, the macroalgae's surfaces were dominated by bacterial amoA genes rather than those from the archaeal counterpart. On the basis of the sequences retrieved from AOB and AOA amoA gene clone libraries from each algal species, the bacterial ammonia-oxidizing communities were related to Nitrosospira spp. and to Nitrosomonas europaea and only 6 out of 15 operational taxonomic units (OTUs) were specific for the host species. Conversely, the AOA diversity was higher (43 OTUs) and algal species specific, with 17 OTUs specific for L. rodriguezii, 3 for O. volubilis, and 9 for P. crispa. Altogether, the results suggest that marine macroalgae may exert an ecological niche for AOB in marine environments, potentially through specific microbe-host interactions.     

CHEMICAL DEFENSE AGAINST HERBIVORY IN THE ANTARCTIC MARINE MACROALGAE IRIDAEA CORDATA AND PHYLLOPHORA ANTARCTICA (RHODOPHYCEAE)

Iridaea cordata (Turner) Bory and Phyllophora antarctica A. et E. S. Gepp (Gigartinales, Rhodophyceae) are common in many high-latitude, shallow benthic marine communities in Antarctica, but previous observations have indicated little or no exploitation by potential herbivores. We have measured retention of test disks over the mouths of the common Antarctic sea urchin Sterechinus neumayeri Meissner, and we have used this assay to examine potential chemical defenses in I. cordata and P. antarctica. Thallus disks of both macroalgal species were retained by the urchins for significantly shorter times than inert filter paper disks and much shorter times than paper disks with a feeding stimulant. Both nonpolar and polar extracts of each macroalgal species added to disks with a feeding stimulant significantly decreased retention times relative to controls. These results support the hypothesis that chemical defenses in I. cordata and P. antarctica play a role in deterring potential herbivores.     

Photosynthetic performance of Arctic macroalgae after transplantation from deep to shallow waters

Transplantation experiments conducted in the Arctic Kongsfjord (Spitsbergen) in summer 1997 investigated the effects of various types of filtered natural radiation (solar, solar without UV-B, solar without UV-A/B) on photosynthesis of various macroalgae. Two brown algal species (Laminaria solidungula, Saccorhiza dermatodea) and four red algal species (Palmaria palmata, Phycodrys rubens, Phyllophora truncata, Ptilota plumosa) were collected from deeper waters, kept in UV-transparent plexiglass tubes wrapped with different spectral cut-off filter foils and positioned at fixed depths in shallow waters for 7-9 days. At regular intervals, chlorophyll fluorescence of photosystem II (optimum quantum yield, F_v/F_m) was determined, as an indicator of photosynthetic performance. The data demonstrate that shallow-water species such as P. palmata are much less affected by natural photosynthetically active radiation (PAR) and UV radiation near the surface than extremely sensitive deep-water species such as Phyc. rubens which exhibited strong decreases in photosynthetic performance, as well as photobleaching of part of the thallus. The other species showed intermediate response patterns. In most species investigated inhibition of photosynthesis was mainly caused by the UV-B wavelengths. Interpretation of the data clearly indicates species-specific tolerances of photosynthesis to ambient solar radiation which can be explained by broad physiological acclimation potentials and/or genetic adaptation to certain (low or high) irradiances. The species-specific photosynthetic performance under radiation stress is in good accordance with the vertical distribution of the macroalgae on the shore.     

Influence of insulin status on extra-mitochondrial oxygen metabolism in the rat.

The effects of alloxan diabetes and subsequent treatment with insulin on extra-mitochondrial oxygen metabolism in terms of D-amino acid oxidase (DAAO), xanthine oxidase and catalase were examined. The DAAO activity in the liver with D-alanine and D-serine decreased by 33-62% in the diabetic group while the decrease in the kidneys was 61-74%. Insulin treatment resulted in overstimulation of DAAO activity in the liver but not in the kidneys. Tissue glycogen content was lowered in the diabetic animals but was restored by insulin treatment. Tissue glycogen content and DAAO activity showed an inverse relationship. The xanthine oxidase activity in the two tissues decreased from 40-55%; the catalase activity decreased from 34-54%. Insulin treatment was unable to restore the xanthine oxidase and catalase activities in both the tissues.     

Long-term changes in the marine macroalgae of Lågskär, Åland Sea (N Baltic)

Species composition, abundance and depth distribution of marine macroalgae were studied at Lågskär, the Åland Sea, in August 1993. Altogether, 26 species were identified and the most diverse vegetation, comprising 18 species, was recorded at a depth of 4–7 m. Comparison with the results obtained by Andersson in 1956 indicates that the total number of species had decreased by 6, and that the best developed and most diverse algal vegetation was then recorded at a depth of 10–15 m. In 1993 species such as Aglaothamnion roseum, Audouinella efflorescens, Phyllophora pseudoceranoides, Halopteris scoparia and Sphacelaria plumigera had disappeared, or were not recorded. Sphacelaria arctica, Cladophora rupestris, Stictyosiphon tortilis and Polysiphonia fucoides , in the 1950s important and dominating species, were greatly reduced in number. However, Pilayella littoralis, Ectocarpus siliculosus, Fucus vesioculosus and Rhodomela confervoides had increased. Compared with the drastic phytobenthic changes recorded in many coastal areas of the Baltic, the changes in the offshore Lågskär area are of minor extent.     

Effects of chelator treatment on aorta and corpus cavernosum from diabetic rats.

Transition-metal catalyzed reactions contribute to oxidative stress, which has been implicated in the pathogenesis of diabetic complications. The aim was to evaluate the effects of treatment with the transition metal chelator trientine on endothelium-dependent relaxation of aorta and corpus cavernosum from streptozotocin-induced diabetes of 8 weeks duration in rats. Effects on cavernosum autonomic innervation were also examined. Diabetes caused a 30.1 +/- 3.8% reduction in maximum aorta endothelium-dependent relaxation to acetylcholine (ACh), which was markedly attenuated (72.7 +/- 10.6%) by trientine treatment. Reversal treatment (4 weeks untreated diabetes, 4 weeks trientine) did not effect endothelium-dependent relaxation compared with aortas from rats with 4 weeks of diabetes, however, there was a 22.5 +/- 6.2% improvement compared with 8 weeks of diabetes. Eight weeks of diabetes caused a 41.5 +/- 6.6% reduction in corpus cavernosum endothelium-dependent maximum relaxation to ACh that was 70.1 +/- 16.9% prevented by trientine. Cavernosum nonadrenergic, noncholinergic (NANC) nerve stimulation caused frequency-dependent relaxation to a maximum of 40.9 +/- 2.4%, which was reduced by diabetes to 24.2 +/- 2.1%. Trientine partially prevented this deficit, maximum relaxation being 31.9 +/- 2.3%. Thus, metal chelator treatment has beneficial effects on aorta and cavernosum endothelium-dependent relaxation and on cavernosum NANC innervation.