Localisation and surface quantification of secondary metabolites in the red alga Delisea pulchra

The localisation of halogenated furanones, the biologically active secondary metabolites from the red alga Delisea pulchra (Greville), was determined by a combination of fluorescence microscopy, culture studies and quantitative chemical analyses. All types of evidence showed that furanones are localised in the central vesicle of gland cells in D. pulchra. These cells release furanones onto the surface of the plant, where they can be quantified using a newly developed surface extraction-technique. Levels of furanones on the surface of the plant were highest near the apical tips (≃100 ng cm⁻²), and decreased towards the base of the alga. Variation in furanone levels within the plant and variation in the number of gland cells followed a similar pattern. The localisation of furanones within gland cells in D. pulchra and the presence and concentrations of furanones on the surface of the plant are consistent with furanones functioning as antifoulants and in mediating other ecological interactions at the surface of the alga. Received: 5 March 1998 / Accepted: 10 September 1998     

Genetic subdivision in the subtidal,clonal sea anemone Anthothoe albocincta

Anthothoe albocincta, a common subtidal anemone along south-eastern Australia, reproduces both sexually through broadcast spawning and clonally through fission. Clones may be distinguished both by their electrophoretic genotypes and the colour of their tentacles and oral discs. Local populations typically consist of many, dense clonal aggregations. However, some clones appear to have locally extensive distributions, forming a series of separate aggregations. The capacity of clones to disperse among local populations is unknown. In this study we used an electrophoretic survey at six allozyme loci to quantify levels of variation among samples from each of 13 local populations and four geographic regions sampled between April 1992 and June 1993. These data revealed that populations of A. albocincta, separated by up to 930 km, were at least moderately subdivided. Levels of geographic variation were high and the average F _ST value (standardised genetic variation) was 0.27 based on genotypes of all individual polyps. This value reflects substantial variation both within (F _PR =0.13) and among (F _RT =0.16) regions. Estimates of gene flow among both neighbouring populations and regions are therefore low (N _e m=1.7 and 1.3, respectively). UPGMA (unweighted pair-group method using arithmetic averages) dendrograms suggest that a genetic discontinuity occurs at the very south-east corner of Australia, paralleling reports for two other south-eastern Australian marine invertebrates. In addition, our analyses and theoretical predictions imply that localised proliferation of clonal genotypes may have caused us to underestimate the potential importance of gene flow via larval dispersal. Moreover, the abundance and vast geographic range of this species suggests that widespread dispersal does occur. Collections from three populations covering the peroid December 1992 to June 1993 were examined by crude dissection along with histological sectioning, and showed A. albocincta to be dioecious, with unisexual clones. Eggs within the ovaries of six females sampled over a 3 mo period were small (96±4 m) and similar to those of related species that produce planktotrophic larvae. In contrast, we found no evidence that clones were dispersed (shared) among neighbouring local populations. An average of only 6% of six-locus genotypes were common to pairs of local populations separated by up to 125 km, this being equal to the percentage expected through sexual reproduction alone. In addition, the percentage of shared genotypes did not decline with increasing geographic separation. These data imply that although asexual reproduction may be used to maintain local populations, the sexual production of genotypically diverse larvae is the primary source of widely dispersed colonists and hence of new clones.     

Photosynthetic carbon acquisition in the red alga Gracilaria conferta

In this continuing study on photosynthesis of the marine red alga Gracilaria conferta, it was found that ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in crude extracts had a K _m (CO₂) of 85 M. Since seawater contains only ca. 10 M CO₂, it appears that this alga must possess a CO₂ concetrating system in order to supply sufficient CO₂ to the vicinity of the enzyme. Because this species is a C₃ plant (and thus lacks the C₄ system for concentrating CO₂), but can utilize HCO₃ ^- as an exogenous carbon source, we examined whether HCO₃ ^- uptake could be the initial step of such a CO₂ concetrating system. The surface pH of G. conferta thalli was 9.4 during photosynthesis. At this pH, estimated maximal uncatalyzed HCO₃ ^- dehydration (CO₂ formation) within the unstirred layer was too slow to account for measured phostosynthetic rates, even in the presence of an external carbonic anhydrase inhibitor. This observation, and the marked pH increase in the unstirred layer following the onset of light, suggests that a HCO₃ ^- transport system (probably coupled to transmembrane H⁺/OH^- fluxes) operates at the plasmalemma level. The involvement of surface-bound carbonic anhydrase in such a system remains, however, obscure. The apparent need of marine macroalgae such as G. conferta for CO₂ concentrating mechanisms is discussed with regard to their low affinity of Rubisco to CO₂ and the low rate of CO₂ supply in water. The close similarity between rates of Rubisco carboxylation and measured photosynthesis further suggests that the carboxylase activity, rather than inorganic carbon transport and intercoversion events, could be an internal limiting factor for photosynthetic rates of G. conferta.     

High-temperature tolerance of photosynthesis in the red alga Chondrus crispus

Chondrus crispus (Stackhouse) is a perennial red seaweed, common in intertidal and shallow sublittoral communities throughout the North Atlantic Ocean. In the intertidal zone, C. crispus may experience rapid temperature changes of 10 to 20C° during a single immerison-emerision cycle, and may be exposed to temperatures that exceed the thermal limits for long-term survival. C. crispus collected year-round at Long Cove Point, Chamberlain, Maine, USA, during 1989 and 1990, underwent phenotypic acclimation to growth temperature in the laboratory. This phenotypic acclimation enhanced its ability to withstand brief exposure to extreme temperature. Plants grown at summer seawater temperature (20°C) were able to maintain constant rates of lightsaturated photosynthesis at 30°C for 9 h. In contrast, light-saturated photosynthetic rates of plants grown at winter seawater temperature (5°C) declined rapidly following exposure to 30°C, reached 20 to 25% of initial values within 10 min, and then remained constant at this level for 9 h. The degree of inhibition of photosynthesis at 30°C was also dependent upon light intensity. Inhibition was greatest in plants exposed to 30°C in darkness or high light (600 mol photons m^-2s^-1) than in plants maintained under moderate light levels (70 to 100 mol photons m^-2s^-1). Photosynthesis of 20°C-acclimated plants was inhibited by exposure to 30°C in darkness or high light, but the degree of inhibition was less than that exhibited by 5°C-grown plants. Not only was light-saturated photosynthesis of 20°C plants less severely inhibited by exposure to 30°C than that of 5°C plants, but the former also recovered faster when they were returned to growth conditions. The mechanistic basis of this acclimation to growth temperature is not clear. Our results indicate that there were no differences between 5 and 20°C-grown plants in the thermal stability of respiration, electron transport associated with Photosystems I or II, Rubisco or energy transfer between the phycobilisomes and Photosystem II. Overall, our results suggest that phenotypic acclimation to seawater temperature allows plants to tolerate higher temperatures, and may play an important role in the success of C. crispus in the intertidal environment.     

Carbon economy and nutrition of the alloparasitic red alga Harveyella mirabilis

Rhodomela confervoides (Huds.) Silva (Ceramiales: Rhodophyta) infected with the alloparasitic Harveyella mirabilis (Reinsch) Reinke (Cryptonemiales: Rhodophyta) was investigated with particular regard to rates of carbon fixation, patterns of soluble and insoluble assimilates, and transfer of reduced carbon from the photoautotrophic basiphyte to the fully heterotrophic epiphyte by means of ¹⁴C-labelling experiments. Gross carbon input by photosynthesis in Rh. confervoides accounted for about 53 g Cd^-1 when based on a biological unit (average individual of 200 mg fresh weight). Some 3% of the remaining net carbon gain (equivalent to about 0.7 g Cd^-1) was transferred from Rh. confervoides to its alloparasite, H. mirabilis (biological unit 0.5 mg fresh weight). Chromatographic examination showed that soluble ¹⁴C was translocated to and stored by H. mirabilis as mannosidoglycerate (=digeneaside) and probably a variety of amino acids. A certain proportion of soluble radiocarbon imported is converted by H. mirabilis to glycerolgalactoside (=floridoside). The bulk of imported ¹⁴C was recovered from the polymeric fraction, mainly from floridean starch.     

Growth of Acropora pulchra

Transplantation experiments were carried out on the branching hermatypic coral Acropora pulchra (Brook) from October 1980 to March 1982 in the northern Philippines. Primary objectives were to determine the effects on the coral of the stresses associated with transplantation, relocation, and the use of different artificial substrates such as flagstones and tires, and to determine possible effects of sedimentation on survival and growth of the transslants. Results of transplantation at different times of the year were also assessed. Comparisons of the responses of transplants with those of undisturbed, control colonies indicated possible significant effects of transplantation tending to retard growth. Sedimentation likewise appeared to suppress growth of the transplants. Growth and survival of the transplants may have been further inhibited by transplantation during the warmer times of the year, and by setting them in prone positions on tires. Effects of relocation at varying distances from source colonies appeared to be negligible.     

Limited change in the diversity and structure of subtidal communities over four decades

A unique archive of photographs from 1969 to 1974 permitted a test of the hypothesis that the diversity and composition of contemporary epilithic communities on subtidal rock walls in the San Juan Islands, WA, USA, has changed over 40 years. Notably, the richness and diversity of sessile taxa was significantly higher in 2008–2011. Furthermore, the multivariate community structure of sessile and mobile taxa differed between the historic and modern eras. Historic communities were characterized by a high percent cover of bare rock and non-calcified algal crusts, consistent with the effects of grazing by chitons and urchins. The rate of sessile community turnover, an index less susceptible to spatial sampling artifacts than richness or diversity, was not significantly different between the two eras. Together with the naturally high spatial variability in these epilithic communities, and the limited replication of historic quadrats, we interpret cautiously the data as evidence of limited change despite a clear shift in temperature and predator (fish) guild composition. The lack of substantial change in rock wall communities may be due in part to their vertical topography, which limits physical disturbance and the preemption of space by weedy algae, two processes that are often associated with “phase-shifts” in other marine ecosystems.     

Genetic diversity and genetic structure in the brown alga Halidrys dioica (Fucales: Cystoseiraceae) in Southern California

Genetic diversity and genetic structure in a population of the brown seaweed Halidrys dioica Gardner were evaluated in five sites in southern California, USA, in 1991, using isozyme electrophoresis. H. dioica is relatively long-lived and has an outcrossing mating system and floating reproductive fronds with the potential for longdistance gamete dispersal. Because these characteristics are hypothetically important in determining genetic diversity and structure, we predicted that genetic diversity would be high and genetic structure would be exhibited only at relatively large geographic scales in H. dioica populations. The data were consistent with the prediction: genetic diversity (% polymorphic loci, no. of alleles/locus, average expected heterozygosity) was high compared to that of other seaweed species. Genetic structure (Wright's F statistics, Nei's genetic distance, point-pattern analysis of alleles) was low within and among distinct rocky reefs over 4 km of coast but high in subpopulations separated by 90 km. Life-history characteristics may be useful predictors of genetic diversity and structure in seaweed populations, but information on selection regimes, long-distance dispersal, and the extent of clonal propagation, for example, are critically lacking.     

The structure of subtidal algal and invertebrate assemblages at the Chatham Islands,New Zealand

We examined the distribution and abundance of organisms on subtidal rocky reefs at nine sites around the Chatham Islands, a remote group 780 km east of southern New Zealand. We sampled five depth strata ranging from 1 to<16 m to identify spatial patterns in the abundance of algae and invertebrates and to assess their variation within and among sites. This information is used to discuss hypotheses concerning community structure at this remote locality. Several patterns were apparent. The immediate subtidal was occupied by the southern bull kelp Durvillaea spp. A suite of 11 fucalean species were dominant to a depth of 10 m with an average abundance of 28 m^-2, while one species, Carpophyllum flexuosum, occurred mostly in deeper water. Only two laminarian species of algae were present at the islands. The indigenous Lessonia tholiformis was abundant at 2.5 to 15 m and was not found in deeper water, while the giant kelp Macrocystis pyrifera was abundant at two sites in 12 to 18 m. The commercially valuable abalone Haliotis iris was extremely abundant in shallow water, with an overall mean of 6 m^-2 at 5 m. The sea urchin Evechinus chloroticus was common, but reached high densities only in small (<25 m²) patches. The characteristic urchin-dominated zones reported in kelp beds world-wide were not seen. There was considerable site-to-site variation in the occurrence and abundance of individual species. Some differences between sites were associated with shelter from swell (e.g. M. pyrifera was found only in sheltered sites) and physical habitat (e.g. juvenile H. iris were found only beneath boulders inshore), but much of the variation could not be explained by physical or depth-related factors alone. We hypothesize that the differences in these kelp bed assemblages compared to mainland New Zealand are partially due to the high degree of endemism at the Chatham Islands. Local variation cannot be explained by herbivory, and is most likely the result of the various life-history characteristics of the major habitat-forming species, the large brown algae.     

Effect of carbon dioxide concentration on calcification in the red coralline alga Bossiella orbigniana

The relationship between various experimental concentrations of CO₂ and calcification in Bossiella orbigniana (Decaisne) was studied by measuring Ca-45 incorporation into the crystalline matrix. Air containing CO₂ at partial pressures (P_{CO 2}) of 0.04 to 5.5% was bubbled through synthetic seawater in incubation vessels. The resultant pH values in the presence of plants ranged from 6.5 to 8.7. The maximum calcification rate appears to lie between 0.11 and 1.05% P_{CO 2}. The data suggest that calcification is controlled by a biological process that may be sensitive to pH and/or to the relative bicarbonate concentration. The data also suggest that a severalfold increase in CO₂ over the present atmospheric level might lead to increased calcification in this marine alga.     

Grazer avoidance may explain the invasiveness of the red alga Gracilaria vermiculophylla in Scandinavian waters

The invasive red alga Gracilaria vermiculophylla has quickly spread across Europe, but it is unclear whether its success is based on a high tolerance to variations in environmental conditions or to the absence of native grazers that feed on this alga. We tested whether native invertebrate grazers prefer native algae to G. vermiculophylla. Feeding preferences of three common herbivores were quantified when offered G. vermiculophylla and native Fucus vesiculosus, Ceramium virgatum, and Ulva intestinalis in no-, two- and multiple-choice trials. Herbivore growth was measured when fed each of the algae separately. Grazers consumed G. vermiculophylla in no-choice trials, but avoided generally this alga when having a choice. U. intestinalis was always preferred over G. vermiculophylla, and grazers fed with U. intestinalis grew faster than those fed with G. vermiculophylla. We conclude that grazers avoid G. vermiculophylla to most native algae, which may benefit G. vermiculophylla in northern European estuaries.     

Studies on digestion and the fine structure of digestive caeca in Eurydice pulchra (Crustacea: Isopoda)

In the carnivorous isopod Eurydice pulchra Leach the mid-gut caeca have a pH of 5.6 and produce a strong acid proteinase, a lipase and a carbohydrase. Electron microscope studies of the digestive caeca of starved and fed animals show that secretion is merocrine and may originate from a complex involving the basal cell membrane and associated mitochondria, producing lysosomes. Absorbed materials, in fed animals, appear to take the form of neutral lipid bodies, glycogen, and proteinaceous crystals, the latter forming within the golgi complex. Digestion of both lipid and protein inclusions was observed to occur intra-cellularly by fusion with free lysosomes. Secretion and absorption may occur in the same cell, and the scheme for the sequence of epithelial cell stages is proposed.     

Long-term variability in the structure of subtidal benthic communities in Puget Sound,Washington, USA

Data on benthic infauna from 4 permanent stations in Puget Sound off Seattle, USA, collected during 1963–1964, 1967, and 1969, revealed considerable stability in numbers of species and specimens and in diversity within stations among sampling dates. The species composition of the faunal assemblages also remained rather constant during the period of investigation, but the relative dominance among the numerically important species varied somewhat. Biomass data did not differ significantly in 1964 and 1969, but the 1967 data were considerably lower at all stations.     

Calcium exchange and the measurement of calcification rates in the calcareous coralline red alga Amphiroa foliacea

⁴⁵Ca washout data of living thalli of Amphiroa foliacea Lamouroux in the light and dark show that there are three kinetically distinct Ca²⁺-exchanging compartments with approximate half-times (t 1/2) of 300, 20 and 3 min. The two slower compartments appear to be exchange from organic Ca²⁺-binding components of the cell wall, while the fast compartment probably represents exchange on the CaCO₃ crystal surface. Killed and decalcified thalli have a fourth compartment, with a t 1/2 of 20 to 35 min (other compartment half-times are 300, 40, 3 min), which has been identified as the greatly increased intercellular space produced during drying and decalcification. The ⁴⁵Ca and ¹⁴C uptake data show that a large proportion of the label initially taken up is into compartments other than the CaCO₃. As a result of this uptake, binding, and exchange of radioisotope, significant errors occur during the measurement of calcification rates, unless a kinetic analysis is carried out. Using such a technique, CaCO₃ calcification rates of A. folicea were measured with ⁴⁵Ca or ¹⁴C as tracers. Light stimulates calcification by up to 2.6 times, depending upon the age of the plant. Young segments have a markedly higher rate of calcification and photosythesis than do the older segments.     

Growth and production of the red alga Gelidium sesquipedale off the Basque coast (northern Spain)

Several aspects of the growth of the agarophyte Gelidium sesquipedale Clemente (Thuret) were studied in sublittoral populations of the Basque coast (northern Spain) from 1981 to 1986. The standing stock of the populations varied greatly with season. During two consecutive years, 60 to 70% of summer biomass had been lost by the end of the unfavourable period of year to growth (autumn and winter). The decline in biomass was mainly due to partial losses of branches, with more branches being lost from larger fronds. Yearly variations in the standing stock were also recorded. Production estimates obtained by a destructive sampling method were not satisfactory because of the spatial heterogeneity of the populations at the three locations studied. Indirect estimation of production by measuring the growth of marked fronds in a control quadrat gave more reliable estimates. Production values of 622±124, 735±147 and 615±123 g dry wt m^-2 yr^-1 were obtained at the three locations, respectively. Frond elongation rates of 7.5, 6.1, 7.5 and 7.9 cm yr^-1, respectively, were recorded during four consecutive years. In winter elongation rates of fronds were rather low. Elongation did not seem to be affected by frond length, the self-shading effect of the canopy, or changes in depth. Cuts resulting from grazing were frequent on fronds through the year, decreasing significantly with increasing depth. Sheared fronds regenerated promptly, independent of season or height at which the cut was made. For commercial purposes the best season for harvesting is summer, which is the period of maximum standing stock, infertility, and calm warm waters that provide easier working conditions for divers.     

Reproductive biology of the red alga Polysiphonia lanosa (Ceramiales) in the Bay of Fundy, Canada

The purpose of this investigation was to establish reproductive phenology of the dioecious marine floridean red alga Polysiphonia lanosa (L.) Tandy. Specifically, the distribution and frequencies of male and female plants, timing of their sexual maturity, fecundity and success of fertilization were investigated. The presence of cystocarps was taken as an indication of syngamy. Populations of P. lanosa in the Bay of Fundy reproduce in spring, summer and autumn. Both male and female filaments were present in equal abundance at all sampling locations, grew in close proximity, and occurred with equal abundance at all elevations above chart datum. During the growing season most of the mature plants contained reproductive organs with apparently functional gametangia containing gametes the structure and behavior of which conformed to those reported in the literature for other species of the genus. Male and female plants of P. lanosa matured synchronously during late May to early July at all collecting sites. The ratio of spermatia to carpogonia calculated for the sampled population was high, 3000–4800 spermatia:1 carpogonium. At the end of the growing season (late October) the fertilization success was at its maximum of 50 to 90% and at its minimum in July at 24%. The three free-living life-history phases (females, males and tetrasporophytes) were abundant and occurred with equal frequency, indicating that a complete life cycle was common at the studied location. Received: 10 December 1996 / Accepted: 16 January 1997     

Sex and life-history stage alter herbivore responses to a chemically defended red alga

Intraspecific variation in resistance to herbivory among genders and life-history phases of primary producers can significantly alter the ecological and evolutionary consequences of plant-herbivore interactions. Seaweeds (macroalgae) with complex life histories have multiple distinct phases with associated variation in traits that can potentially lead to differences in resistance to consumers and provide a unique system in which to simultaneously test the effects of sex and life-history stage on herbivory. We tested the susceptibility to grazing of the three life-history stages and separate sexes of the chemically defended red alga Asparagopsis armata against the sea hare Aplysia parvula, and we related this to the plant quality traits of different stages and genders. Differences in nutrient content and halogenated secondary metabolites between life-history phases were highly sex dependent. Male gametophytes had a low concentration of secondary metabolites and the highest nutrient content. The highest secondary metabolite content was found within the female gametophyte, in the wall of the reproductive structures (cystocarps) that contain the microscopic carposporophyte phase. Feeding choices by A. parvula were consistent with differences in algal quality and defense and resulted in the haploid male gametophytes being the most preferred food type. The diploid carposporophyte found inside the chemically rich cystocarps was the least consumed life-history stage. Selective herbivory of male gametophytes by A. parvula is consistent with an observed shift in gametophyte sex ratio in the field from unity at the beginning of the reproductive season to female bias at the end. The variation in susceptibility to herbivory found between sex and life-history stages of A. armata represents the first example of sex-biased consumption in seaweeds and may contribute to the maintenance of complex life histories such as those found in red algae.     

Multiple trans-Arctic passages in the red alga Phycodrys rubens: evidence from nuclear rDNA ITS sequences

In order to investigate how episodes of geological and climatic change have influenced the distribution and evolutionary diversification of Arctic to cold temperate-North Atlantic seaweed species, intraspecific genetic variation was analyzed among isolates of the sublittoral, benthic red alga Phycodrys rubens (collected between June 1992 and January 1994). Rooted phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer (ITS) sequences and the plastid encoded Rubisco spacer sequences suggest that P. rubens invaded the North Atlantic from the Pacific shortly after the opening of the Bering Strait (3 to 3.5 million years ago), colonizing both the western and eastern Atlantic coasts. Based on these data we further hypothesize that P. rubens survived along the European coasts during the more recent Pleistocene glaciations, while becoming locally extinct along the North American Atlantic coasts. Following retraction of the last ice sheet, the western Atlantic coast was colonized a second time from the Pacific. The presence of two distinct genetic types (based on ITS and Rubisco sequences) along the European coasts is postulated to be a result of isolation and subsequent differentiation. This is likely because ice-free areas are known to have existed in northern Scotland and Norway during the last glaciation. The presence of an East Atlantic genetic type along the West Atlantic coast is believed to be a recent introduction (caused by human activity) of P. rubens to Newfoundland.     

Stable macrofauna community structure despite fluctuating food supply in subtidal soft sediments of Oslofjord,Norway

At a locality at 32-m depth in Oslofjord, Norway, temperature varied from 4.8° to 9.2°C and salinity varied from 31.2 to 33.3 S over a two-year period. There was a peak in chlorophyll a, and C and N in April–June and a smaller peak in November in the sediment. Bacterial numbers showed maxima in July–August and November–December. The macrobenthic fauna was typical of a species-rich and undisturbed boreal community of silt-clay sediments. The community was predominantly composed of surface and subsurface deposit feeders. Over the twoyear period there was little variation in numerical abundance or biomass of the species despite the variation in food input. The lack of seasonality shown by the fauna probably relates to the lack of variability of the physical environment. The mechanism by which this control is achieved, however, is not known. There are large predators/disturbers in the community such as the polychaetes Lumbrineris fragilis, Glycera rouxii, G. alba, Nephthys spp. and the echinoid Brissopsis lyrifera, which probably play an important role in structuring the community.     

Life history of the temperate subtidal gastropod Umbonium costatum

The life history and reproductive biology of the trochid sand snail Umbonium costatum (Kiener) were investigated on a subtidal sandy shore in Hakodate Bay, Japan between 1988 and 1991. Female U. costatum mature at 1 yr of age (shell diameter-11 mm), reproduce twice (in June-July and September-October) in successive years, grow to a maximum size (shell diameter=22 mm) at age 8 yr, increase annual fecundity with age from 2000 (age 1 yr) to 37000 (age 8 yr), and show a maximum monthly gonad somatic index of 8% which is constant among ages. In comparison to a previously studied life history of a tropical Umbonium vestiatium, temperate U. costatum shows more sustained growth and a longer life span after maturation. This could be explained by: (1) the optimal size model concerned with resource investment, in gametes (Sebens 1987); and by (2) bet hedging to compensate large variability in larval success at high latitudes. These two hypotheses are not mutually exclusive, but both are based on season-related extremes of environment at high latitudes where the period suitable for reproduction is short.