Freezing tolerance in the intertidal red algaeChondrus crispus andMastocarpus stellatus: Relative importance of acclimation and adaptation

The effect of repeated daily freezing on photosynthesis, growth and phenotypic acclimation to freezing was studied in the red algaeChondrus crispus Stackhouse andMastocarpus stellatus (Stackhouse in With.) Guiry. Algae used for experiments were collected from Chamberlain, Maine, between March and August 1987, and field observations and experiments were carried out at Chamberlain and Kresge Point, Maine between March 1987 and March 1989. After ca 30 d of daily freezing for 3 h at –5°C photosynthesis ofC. crispus was reduced to 55% of control values. Growth rates ofC. crispus were also reduced in fronds frozen daily compared to unfrozen controls, and eventually fronds became bleached and fragmented resulting in biomass losses. Fronds ofC. crispus, frozen daily, had higher photosynthetic rates following freezing events than unfrozen controls indicating that this species can acclimate to freezing conditions. Acclimation to freezing involves the light-harvesting reactions of photosynthesis. In contrast, photosynthesis and growth inM. stellatus were unaffected by repeated daily freezing for 3 h at –5°C for 36 d. No differences in photosynthesis following freezing were observed between frozen and control fronds suggesting thatM. stellatus does not phenotypically acclimate to freezing. The greater freezing tolerance ofM. stellatus relative toC. crispus results, in part, from genetic adaptations associated with plasma membranes and the light-harvesting reactions of photosynthesis.     

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.     

Photosynthetic response to temperature and desiccation of the intertidal alga Mastocarpus papillatus

When exposed to air during low tide, intertidal macroalgae experience a terrestrial environment and often encounter extreme levels of heating and desiccation. Two aspects of photosynthesis may be influenced by this increase in temperature and decrease in water content during exposure to air: (1) the rate of aerial photosynthesis itself, and (2) the rate at which aquatic photosynthesis recovers upon immersion in water at high tide. This laboratory study examines the effect of air temperature and desiccation on photosynthesis of the intertidal macroalga Mastocarpus papillatus Kützing. Plants were collected at Hopkins Marine Station, California, USA (36°37N; 121°54W) between July and December 1990. When apical tips were exposed to 15 to 25°C air for 2 h, photosynthesis was rapidly recovered upon reimmersion in seawater. Recovery was delayed, but complete, when tissue was exposed to 30°C air, but did not occur after exposure to 35°C air. Desiccation did not influence either the rate or the ultimate level of recovery upon reimmersion. Photosynthesis in air generally decreased with increasing desication, with no net photosynthesis occurring below 25% relative water content. Net photosynthesis of hydrated thalli increased with air temperature from 15 to 30°C, then decreased at 35°C. Dark respiration of hydrated thalli increased over the entire temperature range. This study indicates that thallus heating and desiccation during periods of exposure to air can potentially influence the total carbon budget of M. papillatus.     

Tolerance to freezing and supercooling of interstitial turbellaria and polychaeta from a sandy tidal beach of the island of Sylt (North Sea)

Tolerance experiments on freezing and supercooling (without ice formation) were designed to determine correspondence between tolerance to low temperatures and spatio-temporal distribution of one turbellarian and five polychaete species from sandy beaches of the North Sea island of Sylt. Freezing tolerances are always less than supercooling tolerances. Dinophilus gyrociliatus is significantly more sensitive to freezing (LD₅₀ value after 30 min of freezing:-3°C) than the others, whereas Stygocapitella subterranea is significantly less sensitive (50% mortality at-15.7°C after 30 min). The supercooling tolerances differ considerably among the species. The sequence of tolerances (LD₅₀ values) is as follows: Microphthalmus sczelkowii (-2.9°C after 4 h); D. gyrociliatus (<1 h at-8°C); M. listensis (5.6 h at-8°C); Protodriloides symbioticus (8.2 h at-8°C); Notocaryoplanella glandulosa (66 h at-8°C); S. subterranea (72 h at-8°C). Species of sand flats (d. gyrociliatus, M. listensis, P. symbioticus) have lower tolerances than those of the beach slope (N. glandulosa, S. subterranea). Among the latter, tolerances increase with distance of the distributional area from low tide level. S. subterranea, a species occurring at the uppermost position in the intertidal, proves to be best adapted to both freezing and supercooling. Species preferring deeper regions of the beach (M. sczelkowii) show lower supercooling tolerances than surface dwelling forms. Northern species usually have higher tolerances to cold than southern ones, reaching their distribution limits near the island of Sylt.     

Photosynthesis in whole plants of Chondrus crispus

Photosynthesis in whole plants of Chondrus crispus Stackhouse was measured by ¹⁴C uptake under various light intensities and temperature regimes. In plants collected from waters of 6° to 7°C, photosynthesis did not increase with light intensity over a range of 200 to 2400 foot candles (ft-c). However, in plants collected from waters of 10° to 12°C, the rate of photosynthesis was lower over the range 200 to 1400 ft-c but, at 2000 ft-c, increased to a level similar to that of the 6° to 7°C plants. Ethanol-soluble and insoluble fractions showed similar patterns.     

Freezing rate and duration determine the physiological response of intertidal fucoids to freezing

Differential thermal analysis (DTA) was used to measure the freezing temperature of nine species of red brown intertidal macroalgae from the coast of Maine, USA in 1991. Using slow and rapid cooling rates approximating those found in the field for Ascophyllum nodosum (L.) Le Jol. we found that, for a given rate, the freezing points of all species were similar: -7.06 to -8.02°C for slow cooling (ca. 0.25°C min^-1) and -3.42 to -4.56°C for rapid cooling (ca. 5.0°C min^-1). In the low shore species, Fucus evanescens C. Ag., photosynthesis was inhibited to a greater extent when plants were frozen or thawed rapidly than after slow freezing or thawing. However, in the upper shore species, F. spiralis (L.), photosynthesis recovered rapidly and completely regardless of freezing rate. Rapidly frozen F. evanescens also experienced greater loss of plasmalemmal integrity, evidenced by a greater loss of cellular contents on re-immersion, than those frozen slowly. Light-limited photosynthesis following freezing was more severely inhibited than light-saturated photosynthesis. Respiration was generally enhanced immediately after freezing, but then declined to rates below those of unfrozen controls within 2 h following re-immersion, with control rates of respiration being achieved after a 24 h recovery period. Our data suggest that the physiological consequences of winter emersion at sub-zero temperatures may vary widely between individual plants of freezing-susceptible species, due to the wide variations in freezing rate associated with microhabitat effects.     

Effects of oil pollution on the freezing tolerance and solute concentration of the blue mussel Mytilus edulis

Exposure of blue mussels (Mytilus edulis L.) to oil-polluted sea water for 7 d in the laboratory did not affect their body fluid concentrations of inorganic ions and free amino acids. Mussels exposed to-4°C for 12 h did not freeze, whereas freezing occurred invariably in mussels exposed to-7°C or lower temperatures for the same period. Following freezing at-10°C, oil polluted mussels resumed normal activity considerably more slowly than unpolluted ones, but oil-polluted as well as unpolluted mussels showed normal activity 3 h after thawing. Freezing at-15°C was invariably lethal to individuals of both groups. One possible explanation of the delayed recovery of oil-polluted mussels frozen at-10°C may be that oil components had become concentrated to toxic levels as the amount of solvent water diminished during freezing.     

Photosynthetic studies of Chondrus crispus

The net photosynthesis of intertidal, subtidal, carposporic, tetrasporic, and winter versus summer acclimatized plants of Chondrus crispus Stackhouse were evaluated under different temperatures and quantities of light. The optimum temperature and light conditions for net photosynthesis of C. crispus are seasonally and spatially variable, and there is an adaptive shift in the photosynthetic capacity at different seasons and positions on the shore. Plants collected during the fall and winter had lower light optima (465 to 747 ft-c) for net photosynthesis than spring and summer specimens (about 1000 ft-c). Intertidal populations exhibited a higher rate of net photosynthesis between 250 and 2819 ft-c than subtidal plants. Summer materials have a greater tolerance to high temperatures and a higher temperature optimum than winter materials. Shallow subtidal populations (-6m) exhibited a higher temperature optimum than deep subtidal plants (-12m). Tetrasporic plants (diploid) showed a higher rate of net photosynthesis than carposporic plants (haploid). It is suggested that the diploid plants of C. crispus may extend deeper in the subtidal zone, because they have a higher rate of net photosynthesis than carposporic plants. The results of the present studies are compared with previous physiological studies of C. crispus.Published with the approval of the Director of the New Hampshire Agriculture Experiment Station as Scientific Contribution Number 742.     

Interactions between light and temperature on the physiological ecology ofGracilaria tikvahiae (Gigartinales: Rhodophyta)

Main effects and interactions of light and temperature on rates of growth (), net photosynthesis (P_s), and dark respiration (R) of the red seaweedGracilaria tikvahiae were investigated in outdoor, nutrient-replete continuous-flow seawater culture chambers. Below 15°C,G. tikvahiae did not grow and between 15° and 30°C, both main effects and interactions of light and temperature on and P_s were significant, which explains the occurrence of this alga as a summer annual in its northern range. Temperature interacted with light (I) through its influence on the vs I and P_s vs I curves. The initial slope of the vs I curve, , the light saturation intensity, I_s, and maximum growth rate, _max, were all significantly lowerat 15°C compared to 20°, 25°, or 30°C. Maximum values of _max, the P_s:R ratio and the net photosynthesis:gross photosynthesis ratio (P_s:P_g) all occurred at 25°C, suggesting that this is the best temperature for growth ofG. tikvahiae. Values for P_max increased up to 30°C, indicating that the temperature for maximum growth and net photosynthesis are not the same forG. tikvahiae. Significant photoinhibition of growth and photosynthesis at full incident sunlight (I₀) occurred at 15°C but not at 20°, 25°, or 30°C. Steele's equation fit the 15°C vs I data best, whereas the hyperbolic tangent function fit the 20°, 25°, and 30°C data best. Main effects and interactionof light intensity and temperature on rates of R were also significant (P<0.001). R was highly intercorrelated with and P_s (0.86r0.94), indicating that R inG. tikvahiae is primarily regulated by growth rate and not temperatureper se. Environmental factors that regulate growth, such as light intensity, exert a great influence on R inG. tikvahiae.     

Temperature acclimation of respiration and photosynthesis in the brown algaLaminaria saccharina

Sporophytes of the brown algaLaminaria saccharina (L.) Lamour grown at 15°C contained significantly more chlorophylla (chla) than did similar plants grown at 5°C. The increase in chla in 15°C plants was due to increased numbers of photosystem II reaction centes, and possibly to increased photosynthetic unit size, compared with 5°C plants. These changes were associated with increased values (photosynthetic efficiencies) in 15°C-grownL. saccharina relative to 5°C-grown plants. The changes in together with reduced respiration rates allowed 15°C-grownL. saccharina to achieve net photosynthesis and light-saturated photosynthesis at a lower photon fluence rate (PFR) than 5°C plants when both groups were assayed at the same temperature (15°C). The photon fluence rates necessary to reach the compensation point and achieve light-saturated photosynthesis (I _c andI _k , respectively) increased with increasing incubation temperature inL. saccharina grown at both 5 and 15°C. However, acclimation responses to growth temperature compensated for the short-term effect of temperature onI _c andI _k . Consequently, plants grown at 5 and 15°C were able to achieve similar rates of light-limited photosynthesis, and similarI _c andI _k values at their respective growth temperatures. These responses are undoubtedly important for perennial seaweeds such asL. saccharina, which frequently grow in light-limited habitats and experience pronounced seasonal changes in water temperature.Please address all correspondence and requests for reprints to I.R. Davison     

Sexual and asexual reproduction of Anthopleura dixoniana (Anthozoa: Actiniaria): periodicity and regulation

Reproduction of the sea anemone Anthopleura dixoniana (Haddon and Shackleton) from the high intertidal zone of southern Taiwan (120°41 E; 22°01N) was studied from April 1987 through March 1989. A. dixoniana spawns once a year, in July, and divides asexually by longitudinal fission throughout the year, with a peak in July. During the spawning season, sea anemones>3 mm pedal dise diameter can be sexed, and display a 1:1 sex ratio. Dividing sea anemones are significantly larger than non-dividing individuals, and increase in body size before fission. Under laboratory conditions, individuals kept at 28 C and fed had larger oocytes and a higher division rate than those kept at 18, 22, 25 or 32°C or starved. The division rate significantly influenced the oocyte diameter. The present study revealed for the first time, that a long photoperiod (14 h hight:10 h dark) significantly enhances the growth of oocytes in A. dixoniana under laboratory conditions.     

Population dynamics and reproductive biology of the commensal isopod Colidotea rostrata (Crustacea: Isopoda: Idoteidae)

Colidotea rostrata (Benedict, 1898) is the only known commensal idoteid isopod, living on and mimicking the color of two northeastern Pacific sea urchins of the genus Strongylocentrotus. The population dynamics and reproductive biology of C. rostrata on its host urchins were studied at a low rocky intertidal area in southern California (33°40N; 118°30W) from December 1984 to December 1986. Isopod populations remained relatively stable throughout the 2 yr study, with isopods inhabiting an average of 56.1% of the urchins at 6.1±0.6 (x±2 SE) isopods per urchin. Female isopods reach sexual maturity at a length of 6.8 mm. Fecundity in C. rostrata averaged 11.8±0.9 (x±2 SE) embryos per brood, and increased with female body size. Breeding occurs all year long in C. rostrata, with a main reproductive period between the warmer spring and summer months. Newly released mancas and juvenile isopods were present during all months of the study. C. rostrata differs from the free-living Idoteidae in its smaller maximum size, reduced fecundity, 1:1 sex ratio, and low juvenile mortality. These features may represent adaptations to a commensal life style that reflect a reduced mortality pressure on these isopods.     

Effect of phosphate and ammonium levels on photosynthetic and respiratory responses of the red alga Gracilaria verrucosa

Gracilaria verrucosa (Hudson) Papenfuss exposed to nutrient enriched media (0.1 mM PO₄; 1.0 mM NH ₄ ⁺ ) by pulse feeding 2 h every third day for a period of 5 wk at 20°C and 25–30 salinity showed significantly higher rates of photosynthesis regardless of photon flux density correlated with increased pigment levels. Algae in nonenriched media showed significantly higher levels of soluble carbohydrates and decreased levels of phycoerythrin and chlorophyll a. Photosynthetic and respiratory responses to temperature 15°, 25°, 30°C and salinity (15, 25, 30 S) combinations indicate broad tolerances by both nutrient enriched and non-nutrient enriched algae. Photosynthetic and respiratory rates were highest at the high temperatures. Pulse-fed algae had significantly higher photosynthetic rates than non-nutrient enriched plants at all temperature and salinity combinations. Non-nutrient enriched algae had significantly higher respiratory rates than nutrient enriched algae at only 30°C and 15. The respiratory rates of both nutrient enriched and non-nutrient algae decreased under combinations of higher temperatures and salinities. G. verrucosa, grown without nutrients, has lower tolerances to environmental stresses.     

Temperature-influenced infection rates in the Chondrus crispus—Petersenia pollagaster pathosystem: a regression analysis

Cross-infection experiments were performed to determine the influence of temperature on infection rate in the Chondrus crispus Stackhouse-Petersenia pollagaster (Petersen) Sparrow pathosystem. C. crispus thalli were collected at Pubnico Harbor, Nova Scotia, Canada in the fall of 1981 to 1984. Infective zoospores were used to inoculate healthy thalli at five different temperatures. The highest infection rate was obtained at 20°C, while significantly lower rates were obtained at temperature extremes. The parasite's life cycle, consisting of infection of healthy thalli, endobiotic development, and release of zoospores, was completed in 48 to 72 h at 15° to 20°C.     

Response of gametophytes of Ecklonia radiata (Laminariales) to temperature in saturating light

Gametophytes of Ecklonia radiata (C.Ag.)J.Ag. from two New Zealand locations with different field temperature ranges were exposed to temperatures of 5° to 26°C in saturating light. Plants from Goat Island Bay (Lat. 36° 16S, Long. 174°48E) grew in 9.3° to 25°C and reproduced in 9.3° to 24°C. There was no growth at 8°C and plants died at 26°C. Plants from the cooler location, Houghton Bay (Lat. 41°20S, Long. 174°40E), grew from 8° to 24°C and reproduced up to 15°C but not at 21.5°C. The plants did not grow at 6°C and died at 26°C. The timing of the first cell division and subsequent growth rate were retarded close to the upper and lower tolerance limits. Reproduction was a broad optimum of roughly 12° to 20°C. Within this range, fertile female gametophytes grown at lower temperatures had fewer, larger cells and thus fewer potential ova than those grown at higher temperatures.     

Influence of high temperature and residual chlorine on marine phytoplankton

Marine phytoplankton forms are frequently exposed to sudden biological changes such as rapid rise in water temperature and chlorine content of their environment, resulting from the use of sea water for cooling purposes by electric generators. The direct influence of these effluents, i.e. inhibitory effects of high temperature and residual chlorine on growth and photosynthesis of Chlamydomonas sp. and Skeletonema costatum, were investigated experimentally. Chlamydomonas sp. and S. costatum exposed to high temperatures were affected in their growth from 43° and 35°C, respectively, by immersion of the respective cultures in a warm bath for 10 min. Treatment at high temperatures of 40 °C and 30° 35°C for 10 min, influenced their photosynthetic activities, which were completely inhibited immediately after 10 min exposure at 42° and 37 °C, respectively. S. costatum was killed by chlorine at a concentration of 1.5 2.3 ppm when exposed for exactly 5 or 10 min, while Chlamydomonas sp. was not irreversibly damaged even at 20 ppm chlorine or more with the same exposure period. These results lead to the conclusion that the high temperature of, and residual chlorine in, effuents from a power plant discharging into the open sea, should not cause great damage to marine phytoplankton in that area.     

Autecology and clonal variability of the marine centric diatom Thalassiosira rotula (Bacillariophyceae) in response to light,temperature and salinity

The influence of 49 combinations of salinity (10–40 S, at 5 S intervals) and temperature (0°–30°C, at 5C° intervals) on the maximum daily division rate (K) and 18 combinations of light intensity (six levels) and temperature (5°, 15°, and 25°C) on photosynthesis, cell division, and chlorophyll a was examined using two clones of Thalassiosira rotula Meunier isolated from the upwelling area of Baja California (clone C8) and from Narragansett Bay, Rhode Islands (clone A8). Physiological differences appear to characterize these to clones with regard to their temperature tolerance (C8 5°–30°C, A8 0°–25°C), maximum growth rate (C8 K=2.9, A8 K=2.4), chlorophyll a content, and in the rates of growth and photosynthesis in response to light intensity and temperature. Optimum salinity for both clones (25–30 S) was generally independent of temperature, while chlorophyll a content decreased with temperature. T. rotula is a cosmopolitan paractic species; experimental studies indicate that it is eurythermal and moderately euryhaline. Comparison of five additional Narragansett Bay isolates of T. rotula reveal minimal spacial or temporal variability in genetically determined physiological characteristics within this local population.     

Comparison of three techniques for administering radiolabeled substrates to sediments for trophic studies: uptake of label by harpacticoid copepods

Microbial grazing by two species of meiofaunal harpacticoid copepods (Heteropsyllus nunni and Thompsonula hyaenae) was determined by uptake of radioactive labels following their introduction into natural sediments from a low-energy intertidal site in Florida (29°5440N; 84°3130W) in May 1986. Grazing was related to three methods of radioactive-label introduction: injection, porewater replacement, and slurry. Uptake of label by harpacticoids was examined using two dual-label combinations, ³H-thymidine/¹⁴C-bicarbonate and ³H-thymidine/¹⁴C-acetate. The injection and porewater-replacement methods yielded statistically indistinguishable results. Results obtained by the slurry method differed significantly from the other two methods. We suggest that the unique results obtained in the slurry method were a consequence of the disruption of microbial-meiofaunal spatial relationships.     

Algal overgrowth of alcyonacean soft corals

Colonies of the soft coral Lobophytum pauciflorum (Ehrenberg, 1834) (Coelenterata: Octocorallia: Alcyonacea: Alcyoniidae), some of which were heavily overgrown by the algae Ceranium flaccidum (Kuetzing) Ardissone and Enteromorpha sp., and other minor epizoites, were collected at Taylor Reef (17°50S; 146°35E) in the Great Barrier Reef. Overgrown colonies contained the diterpene 2-epi-sarcophytoxide as the major secondary metabolite, while conspecific colonies with clean polyparies contained two diterpenes in approximately equal amounts: 14-hydroxycembra-1,3,7,11-tetraene and 15-hydroxycembra-1,3,7,11-tetraene. By contrast, twenty conspecific pairs of overgrown and clean colonies of other alcyoniid soft corals collected from Pelorus Channel, Palm Island Group (18°34S; 146°29E), showed no chemical differences within in the pairs. Cultures of a common species of Ceramium [C. codii (Richards) Mazoyer] were incubated with different concentrations of nine soft-coral-derived diterpenes and significant algal growth inhibition was observed in many cases. It appears that terpenoids from soft coral may contribute to the lack of epizoic organisms on soft-coral polyparies.     

Photosynthesis and respiration in the alga Ahnfeltia plicata in a flow-through system

Photosynthesis and respiration in Ahnfeltia plicata (Huds.) Fries (Gigartinales) was measured in a seawater flowthrough system at different temperatures, salinities and photon flux densities (PFD). The exchanges of dissolved oxygen and inorganic carbon were continuously recorded with an oxygen probe and a pH electrode measuring variation in CO₂–HCO ₃ ^- equilibrium as pH changes. Highest apparent photosynthesis at moderate photon flux density (PFD 50 E m^-2 s^-1) was found at 15°C and 33 S. Photosynthesis was measured up to PFD 500 E m^-2 s^-1 and no light saturation was documented. In the present experimental set-up, with continuous supply of fresh seawater, the number of limiting factors during photosynthesis measurements is reduced.