Effect of epiphytism on reproductive and vegetative lateral formation in the brown, intertidal seaweed Ascophyllum nodosum (Phaeophyceae)

In the brown alga Ascophyllum nodosum (L) Le Jolis, a common species on sheltered Northern temperate rocky shores, gametes are produced in receptacles that emerge from small depressions (lateral pits) along the branched frond. These lateral pits are also the preferred settling site for the obligate epiphyte Polysiphonia lanosa (L) Tandy. Therefore, epiphytism can be expected to interfere with host reproductive output. The present study investigated the potential impact of the epiphyte on A. nodosum in two series of laboratory experiments that measured: (i) the direct shading of the host plant underneath an epiphyte canopy; and (ii) the development of receptacles in clean and epiphytised A. nodosum segments (excised from individual fronds) over a 6 month period. These experiments showed that light reaching emerged fronds underneath a dense epiphyte cover was reduced by 40%, and this was independent of the degree of desiccation the epiphyte experienced. Concurrently, in the growth study with epiphytised A. nodosum segments (segments with one clean and one epiphytised lateral pit) total receptacle biomass per epiphytised fragment was significantly reduced compared with clean segments (0.52 g and 1.25 g per gram of frond segment, respectively), although this effect was only significant in A. nodosum from sheltered shores. However, expressed as biomass per lateral pit, receptacle biomass in the remaining clean lateral pits in epiphytised segments was significantly increased in segments from both shores, demonstrating that A. nodosum can at least partially compensate for the loss of production resulting from epiphytism.     

SETTLEMENT AND SURVIVAL OF POLYSIPHONIA LANOSA (CERAMIALES) SPORES ON ASCOPHYLLUM NODOSUM AND FUCUS VESICULOSUS (FUCALES)1

The settlement patterns of spores of Polysiphonia lanosa (L.) Tandy on Ascophyllum nodosum (L.) Le Jolis and Fucus vesiculosus L. were studied using a flow tank. Settlement sites were defined as ‘sheltered’ or ‘exposed.’ Surface area calculations revealed non-random settlement on A. nodosum, with higher than expected spore frequencies on the thallus and lateral pits and lower than expected frequencies on the vesicles. Settlement of F. vesiculosus was random and significantly lower than on A. nodosum. On the shore, survival of sporelings from September (post-sporulation) to May (pre-sporulation) was highly non-random on both basiphytes. On A. nodosum, lateral pits ('sheltered') showed the highest survival frequency. Here the proportion of surviving sporelings increased over the study period, whereas the proportion on open thallus area ('exposed') decreased. On F. vesiculosus also preferential survival occurred on ‘sheltered’ sites such as vesicle/thallus interfaces and wounds. Between September and May, all P. lanosa sporelings were lost from ‘exposed’ areas (thallus surface and vesicles). Overall, frequencies of surviving sporelings were much greater on A. nodosum than on F. vesiculosus. These results are discussed with reference to basiphyte morphology, epiphyte removal mechanisms and the survival stratagy of P. lanosa.     

COLONIZATION AND GROWTH OF ASCOPHYLLUM NODOSUM (PHAEOPHYTA) IN MAINE1

Colonization and growth of Ascophyllum nodosum (L.) Le Jol. were investigated in two Maine estuaries from 1972 to 1978. Following denudation of intertidal rock, substrata were initially colonized by Fucus vesiculosus L.; eventually, Ascophyllum supplanted Fucus, and became dominant in terms of percentage cover. Ascophyllum settled first and most densely in the low intertidal zone, but its fastest growth occurred in the mid-intertidal zone. Some, but not all, Ascophyllum germlings produced a vesicle within one year of colonization. The mean annual growth of A. nodosum was variable among sites, zones and years.     

PHYSIOLOGICAL AND POPULATION ECOLOGY OF INTERTIDAL AND SUBTIDAL ASCOPHYLLUM NODOSUM (PHAEOPHYTA)1

Morphological, demographic and physiological characteristics of Rhode Island intertidal and subtidal populations of Ascophyllum nodosum (L.) Le Jolis were compared in order to examine factors influencing vertical distribution. The two populations had distinctive morphologies: subtidal plants were narrower (more terete) and highly branched compared with intertidal plants. The subtidal population showed signs of necrosis and breakage, which was reflected in significantly shorter mean plant size. High survivorship and low recruitment of both population resulted in relatively constant densities, averaging 91 and 50 plants per m² in the intertidal and subtidal habitats, respectively. Intertidal plants had higher mean annual growth rates (25 cm.yr^?1) than subtidal plants (2 cm.yr^?1). In general, intertidal plants had higher photosynthetic capacity and nutrient (NO₃^?) uptake rates than the subtidal population but maintained lower light-harvesting pigment and tissue nitrogen concentrations. Although Ascophyllum nodosum is capable of survival and growth in subtidal as well as intertidal areas, results of this study suggest that different selective pressures affect persistence in each habitat. The scarcity of plants in the subtidal environment may be due to the lack of a critical balance between algal production, allocation of photosynthate, and the negative effects of grazers or competitors.     

Experimental oil exposure of Ascophyllum nodosum (L.) Le Jolis

The effect of weathered, crude oil on Ascophyllum nodosum was tested in an outdoor experiment and in a laboratory experiment. Adult A. nodosum plants which were exposed to oil in an outdoor basin for 3 days showed reduced growth for a certain period of time, after which growth recovered. Also, some necrotic damage was observed. Three-week-old and 1-year-old germlings placed under the adult plants showed no significant reduction in number compared to control plants. In the laboratory experiment some of the 1-month-old germlings were initially exposed to oil for 6 min, some for 1 h, some for 3 h and some for 5 h. After 3 weeks in running, filtered water oil-exposed germlings had reached the same length as those in the control groups, and no effect of exposing the germlings to oil for different lengths of time at the start of the experiment was found. The results indicate that a limited oil pollution of weathered, crude oil may reduce growth and biomass of adult plants for a certain period of time. However, this need not be detrimental to a population with good recruitment from germlings or vegetative shoots growing under a dense canopy layer.     

Adaptations by macroalgae to low carbon availability. I. A buffer system in Ascophyllum nodosum, associated with photosynthesis

Abstract The exchange of CO₂, H⁺ and O₂ between seawater and the intertidal brown macroalga Ascophyllum nodosum (L.) Le Jolis were measured in a flowthrough system. While the algae were kept in darkness, seawater with artificially increased alkalinity and pH at 9.85, was alternated with ‘normal’ seawater at pH 8.0. A proton buffering system, with capacity to release and reabsorb about 20 μmol protons per gram alga (fresh weight) was revealed. As the algae were returned to the ‘normal’ seawater, the kinetics of proton reabsorbtion indicated that a proton uptake was gradually induced. This proton uptake, which was not connected to ion exchange in the cell wall, reached its maximum after 12 h. If subjected to high alkalinity seawater in the light, A. Nodosum for a certain period of time was capable of carrying out O, evolution in excess of the import of inorganic carbon. This ‘photosynthetic buffering capacity’ amounted to about 17 μmol O; per gram alga. Besides depending on a buffer of photorcducible substances, this ‘photosynthetic buffering capacity’ appeared to be functionally connected with the proton buffer. The time course for the discharge of the ‘photosynthetic buffer system’ and for the reabsorbtion of protons into the proton buffer (about 6h for 90× of the capacity at a temperature of 6°C) suggests that the ‘photosynthetic buffer system’ has a functional importance in the adaptation of A. nodosum to intertidal regions. The function of the buffer system is discussed in relation to the crassulacean acid metabolism (CAM)-like characteristics recently shown for the intertidal brown algal family Fucaceae.     

A MODEL FOR GAMETE RELEASE IN ASCOPHYLLUM NODOSUM (PHAEOPHYTA)1

The question of what controls gamete release in Ascophyllum nodosum (L.) Le Jolis was studied at six sites along the central coast of Maine. Percent release was assessed weekly along randomly marked transect lines in the mid-intertidal zone. Six independent variables–water temperature at high tide, air temperature at low tide, nitrogen (ammonia, nitrite, and nitrate), and salinity–were measured concurrently. Stepwise multiple regression analysis on the percentage of plants having released gametes revealed that water temperature at high tide accounted for most of the among-site variation (R²= 0.77) in the timing of release. The addition of Julian day increased the R² to 0.82; no other variables were significant. Probit analysis, based on water temperature at high tide, generated an environmentally realistic model for predicting gamete release. The model predicts the onset, midpoint, and termination of gamete release at 6, 10, and 15°C, respectively, and the midpoint at a cumulative water temperature of 358°C. This model has value for developmental studies and, potentially, for reseeding A. nodosum populations. Probits may be useful for characterizing phenological events in other fucoids and algal species.     

PHOSPHOENOLPYRUVATE CARBOXYKINASE ACTIVITY IN ASCOPHYLLUM NODOSUM (PHAEOPHYCEAE)1

PEP-dependent⁴ CO₂-fixation by extracts of Ascophyllum nodosum (L.) Le Jol. is reported. The carboxylation of PEP is Mn²⁺ dependent and ATP is shown to be a product. IDP was found to be less efficient as a phosphate acceptor than ADP and 3-mercaptopicolinic acid inhibited the carboxylation reaction. Extracts decarboxylated OAA only in the presence of ATP and had high activities of MDH and GOT. This evidence, together with the probable absence of PEPC, PEPCTrP, and PC in A. nodosum extracts, favors the view that PEPCK is responsible for the light-independent CO₂-fixation observed in this alga.     

SEQUENCE AND PHYLOGENY OF THE PSAB GENE OF PYLAIELLA LITTORALIS(PHAEOPHYTA)1

The psaB gene codes for one of two highly conserved P700 chlorophyll a apoproteins of photosystem I. This gene was cloned from the brown alga, Pylaiella littoralis (L.) Kjellm., and its primary sequence was determined. The inferred amino acid sequence of the P. littoralis protein was compared to homologous sequences from land plants, green algae, and a cyanobacterium. The psaB protein sequence is very conserved in all the examined taxa, and an unrooted phylogenetic tree, generated from a distance matrix, shows that the P. littoralis gene is closer to that of the cyanobacterium Synechocossus sp. PCC 7002 than are those of green algae, land plants, and Euglena gracilis.     

Effects of Tasco (a seaweed extract) and heat stress on N metabolism and meat fatty acids in wether lambs fed hays containing endophyte-infected fescue

Wether lambs (n = 27, average BW = 40 kg) were used to test response to forage treated with Tasco-Forage (an extract of the brown kelp Ascophyllum nodosum) prior to conserving, or to direct feeding of the extract (Tasco-EX). Hays made from endophyte (Neoyphodium coenophialum)-infested tall fescue (Festuca arundinacea)-based pasture received 0 or 3 kg of Tasco/ha prior to harvest. Lambs, blocked by weight, were randomly allotted to three diets: (1) control hay, (2) treated hay, and (3) #1 + Tasco-EX fed at 1% of the diet. Hays were low in CP (<7%) so all lambs were fed soybean meal (12% of the diet) in addition to trace mineralized salt. Diets were fed at 1.5% BW to prevent refusals. Total collections (7 d) were made during periods without or with applied heat stress. After each period, rumen contents were obtained to determine pH, NH₃ and VFA. Lambs were sacrificed post-trial. A subset was used to evaluate sensory traits and muscle fatty acids. Lambs were in negative N balance during the study and Tasco treatments did not affect N metabolism. Fecal N tended (P < 0.10) to increase with short duration heat stress causing a concomitant decrease (P < 0.05) in apparent N digestibility (58.6 versus 56.1%; S.E. = 0.7). Urinary N loss decreased (P < 0.001) with heat stress (8.0 versus 5.9 g/d; S.E. = 0.2), resulting in increased (P < 0.001) N retention (−2.1 versus −0.3 g/d; S.E. = 0.2). Apparent OM digestibility was not affected by heat stress but was greater (P < 0.05) for lambs fed Tasco-EX treatment than those fed treated hay. Treatment diets decreased (P < 0.05) ruminal butyrate. Heat stress increased (P < 0.05) acetate and total VFA and decreased (P < 0.01) ruminal pH. A tendency (P < 0.11) of increased 14:1ω5, decreased (P < 0.05) 18:0 and total saturated fatty acids in muscle was observed with Tasco diets. Meat sensory characteristics were not affected by treatment. Tasco may alter some aspects of rumen or lipid metabolism but has no effect on N metabolism or meat sensory characteristics of sheep fed restricted, low-quality diets.     

Studies on Isoleucyl-tRNA Synthetase of an Extreme Thermophile,Thermus thermophilus HB8

Isoleucyl-tRNA synthetase (IRS) was partially purified from an extreme thermophile, T. thermophilus HB 8. The molecular weight (11.5 ×10⁴) and some kinetic constants were obtained and compared with IRS from other sources.

The present IRS catalyzed both isoleucine dependent and valine dependent ATP-PPi exchange reactions (optimum at around 80°C) but not valyl-tRNA formation. The optimum temperature for isoleucyl-tRNA formation was 62°C with E. coli tRNA and 75°C with T. thermophilus tRNA.

The enzyme showed a remarkable thermostability. The addition of E. coli or T. thermophilus tRNA enhanced the thermostability of the enzyme, which was shown to be fully active up to 77°C. When E. coli tRNA was used, the loading activity decreased in parallel to the unfolding of the substrate tRNA molecule. From these results the relation is discussed between tRNA conformation and function.     

PROPERTIES OF PHOSPHOENOLPYRUVATE CARBOXYKINASE FROM ASCOPHYLLUM NODOSUM (PHAEOPHYCEAE)1

Phosphoenolpyruvate carboxykinase was purified to electrophoretic homogeneity from the brown alga. Ascophyllum nodosum (L.) le Jol. Its molecular mass was 60 kDa as determined by gel filtration. The pH optimum of the carboxylating reaction was 7.9 and the apparent K_m for PEP⁴, ADP and HCO₃^- were 0.036, 0.0116 and 50 mol · m^-3, respectively. Rates of light and dark carbon fixation are also reported for A. nodosum apices and it is shown that rates of in vitro PEPCK activity would account for the observed rate of dark fixation. The physiological role of PEPCK is discussed in relation to light-independent carbon fixation.     

BENTHIC METABOLISM ON A SHELTERED ROCKY SHORE: ROLE OF THE CANOPY IN THE CARBON BUDGET1

While the importance of canopy‐forming algae in structuring ecosystems is recognized, their role in the carbon budget is still not well understood. To our knowledge, no measurements of rocky shores primary production and respiration under emersion periods have been carried out in situ. A benthic chamber coupled to a CO₂‐infrared gas analyzer was used to measure gross primary production and respiration on the Ascophyllum nodosum (L.) Le Jol. zone of a sheltered rocky shore in Brittany, France. Over a year of monthly measurements on the zone with and without the A. nodosum canopy showed fairly high production and respiration values for the global community as well as carbon fluxes due to the canopy that largely dominated the benthic metabolism of the zone. The strong canopy respiration relative to the primary production also suggested a high metabolic activity by microscopic heterotrophs on the surface of the alga. Both the canopy and the understory annual primary production and respiration were under the control of light and temperature seasonal variations. Finally, the range of the amount of carbon produced on the A. nodosum zone during diurnal emersions was estimated. Additional measures accounting for the day–night cycles and seasonal light variations over an entire tidal cycle are, however, necessary to establish an annual carbon budget. Such measures using the benthic chamber together with complementary techniques would allow a better understanding of the functioning of sheltered rocky shores.     

Water Status Changes in Shoots of a Seaweed Ascophyllum nodosum at Subzero Temperatures

The gradient freezing and NMR spectroscopy were used to study the physical state of water in apices of the intertidal seaweed Ascophyllum nodosum at freezing temperatures. In the apices exposed to temperatures below –10°C, two fractions of bound water were revealed. The slow (T₂ 50 ms) fraction of bound water was completely frozen at –25°C, and its freezing rate was temperature-sensitive. This fraction was apparently associated with protoplasmic water and cell-wall polysaccharides. The fast fraction (T₂ < 10 ms) of bound water was presumably due to water-soluble globular proteins. The freezing rate for this fraction depended on neither the temperature nor the amount of water. The presence of unfrozen water in apical cells at –40°C was demonstrated. The role of this water fraction in maintaining the native structure of biomacromolecules and apex survival is discussed.