Resting spore formation of the marine planktonic diatom Chaetoceros anastomosans induced by high salinity and nitrogen depletion

The marine planktonic diatom Chaetoceros anastomosans, which was isolated from Sagami Bay, was used for a study of resting spore formation mechanisms in batch culture experiments. Vegetative cells could grow at salinities ranging from 20.7 to 45.5‰, and resting spore formation was enhanced significantly in nitrate-depleted, high salinity media (40.0 to 45.5‰). The rate of resting spore formation (1.9 d⁻¹) was comparable to the specific growth rate (1.8 d⁻¹) of vegetative cells in the exponential growth phase in normal salinity medium. The size of resting spores formed under high salinity conditions was smaller than that of spores formed in normal salinity media. Unlike vegetative cells, resting spores seemed to possess some mechanisms to survive over a wider range of salinities by resisting bacterial attacks on their cell walls. Received: 4 August 1996 / Accepted: 27 August 1996     

Survival and recovery of resting spores and resting cells of the marine planktonic diatom Chaetoceros pseudocurvisetus under fluctuating nitrate conditions

Of the two resting life-forms of the planktonic diatom Chaetoceros pseudocurvisetus Mangin formed during periods of nitrate depletion, resting spores survived at least 1 month after spore formation at 24 °C, while resting cells survived only for about 10 d at the same temperature. Under nitrogen limitation, resting cells exhibited higher specific death rates than resting spores at temperatures ranging from 5 to 30 °C. After nitrogen replenishment, resting spores required a certain lag period of about 1 d to initiate vegetative growth at levels of nitrate supply from 0.5 to 20 M, while resting cells initiated vegetative growth almost immediately. Resting spores exhibited an intracellular accumulation of the supplied nitrate during germination and initial vegetative growth. The resting cells, however, exhibited more active vegetative growth, closely coupled with the uptake of the supplied nitrate. The resting spores and resting cells appear to play different roles in the maintenance of populations under nutrient fluctuations depending on the interval length between nutrient fluxes in natural waters. Received: 27 April 1998 / Accepted: 1 March 1999     

Resting spore formation and phosphorus composition of the marine diatom Chaetoceros pseudocurvisetus under various nutrient conditions

The marine diatom Chaetoceros pseudocurvisetus was used for a study of resting spore formation and cellular phosphorus composition. Resting spores were found in any culture medium with ample silica, including nitrogen limited, phosphorus limited and nutrient replete conditions. Resting spores protected themselves with thick silica walls, so that vegetative cells required about 3 pmol cell^-1 of additional silica to form resting spores. Phosphorus compounds in the cells were divided into eight fractions: nucleotide-P, orthophosphate, acid soluble polyphosphate, sugar phosphate, nucleic acid-P, acid insoluble polyphosphate, lipid-P and residual-P. The sum of orthophosphate, sugar phosphate and nucleic acid-P comprised over 65% of the total phosphorus in cells under any culture conditions. Sugar phosphate was the most variable component, being most abundant in vegetative cells and least abundant in resting spores.     

Life-form population responses of a marine planktonic diatom,Chaetoceros pseudocurvisetus,to oligotrophication in regionally upwelled water

Life-form population responses of a centric planktonic diatom,Chaetoceros pseudocurvisetus Mangin, were investigated in summer 1986 and 1988 in the Izu Islands, Japan, in regionally upwelled water where nutrient concentration changed from favorable to unfavorable conditions for active growth and reproduction (oligotrophication). Two types of life form were observed: vegetative cells of healthy and unhealthy looking conditions and resting spores. The observed life-form responses were experimentally evaluated along with a depletion of limiting nutrients. The algal population ceased vegetative growth and initiated resting spore formation with a disappearance of limiting nitrate, and this life-form response seemed to be triggered by the decrease of cellular nitrogen content below a certain level. Since a large amount of silicon was required for the resting spore formation, a part of vegetative cells were unable to form resting spores and formed unhealthy looking vegetative cells under insufficient concentrations of silicic acid. Percentage shares of the resting spores in the population were linearly related to the amounts of available silicic acid. Vegetative cells which did not form resting spores showed greater mortality than resting spores by attack of bacteria and protozoa; however, vegetative cells could respond quickly to possible nutrient replenishment.     

Sexuality and cyst formation of the spring-bloom dinoflagellate Scrippsiella hangoei in the coastal northern Baltic Sea

The temporal sequence and the magnitude of the sexual reproduction and subsequent cyst deposition of the common spring-bloom dinoflagellate Scrippsiella hangoei (Schiller) Larsen was studied during spring 1996 on the SW coast of Finland, Baltic Sea. The abundances of the different size of fractions of S. hangoei (14 to 18 μm, 18 to 22 μm and >22 μm) were monitored in the water column, and the deposition of resting cysts was measured using moored sediment traps. Cyst sedimentation rates were measured throughout the seasonal cycle in order to estimate cyst resuspension rates for the quantitative assessment of the fraction of population undergoing encystment. The onset of sexual reproduction, indicated by a significant increase of the small cells (14 to 18 μm) representing gametes, occurred in a nutrient replete environment well before the exponential growth phase and peak abundances of vegetative cells. Gamete formation was followed by high abundances of large cells (>22 μm) representing planozygotes, and subsequent sedimentation of resting cysts. Approximately 60% of the asexually growing bloom population was estimated to form planozygotes, suggesting that encystment was an important factor in bloom termination and possibly plays a role in the regulation of the magnitude of the bloom. Finally encystment accounted for 40% of the entire S. hangoei population, resulting in a considerable loss of the bloom population and an input of the vernal phytoplankton biomass to the benthos. Received: 11 December 1998 / Accepted: 8 April 1999     

Cell-cell recognition system in gorgonians: description of the basic mechanism

A comparative investigation of the chemical composition of Thalassiosira antarctica var. antarctica vegetative and resting stages revealed C:N and C:chl a ratios to be lower in vegetative cells. These trends primarily reflect vegetative levels of C/cell below, and N/cell and chl a/cell levels above those of spores. There was a change in chemical composition with the initial formation of resting spores, and spores continued to modify their composition while maintained in a cyclic light/dark regime for about one and one half weeks. Most notable was a net increase in carbon and chlorophyll a per cell. Spores then subjected to darkness for over one week appeared to retain most of the carbon and chlorophyll a previously synthesized. These findings support the idea that resting spores enhance the survival capabilities of a species under adverse conditions.     

Feeding and bacteriolytic responses of the deposit-feeder Abarenicola pacifica (Polychaeta: Arenicolidae) to changes in temperature and sediment food concentration

Deposit-feeders can respond to seasonal fluctuations in food concentration both functionally (e.g. by adjusting feeding rates) and physiologically (e.g. by changing the concentration of bacteriolytic agents in gut fluids). Laboratory feeding experiments were carried out (11 to 21 July 1997) with the arenicolid polychaete worm Abarenicola pacifica (Healy and Wells). Objectives were to test for separate and interactive effects of sediment food concentration and temperature (6, 11, and 16 °C) on deposit-feeder functional (feeding rates) and physiological (bacteriolytic activity of gut fluids) responses. Food concentration was varied experimentally using sieved (1 mm) natural sediments (Md φ=2.00; 0.6% organic) mixed with combusted (500 °C, 8 h) sediments for final concentrations of 25, 50, and 100% natural sediment. Sediment food quality was measured as: (1) bioavailable amino acids (EHAA), (2) chlorophyll a (chl a), and (3) bacterial abundance. Feeding rates were inferred from egestion rates (ER, g h⁻¹) and analyzed with respect to worm size. Bacteriolytic activity of midgut fluids was assayed turbidimetrically against two bacterial isolates, after worms had fed on experimental sediments for 15 d. Temperature and food concentration both significantly affected feeding rates, with maxima occurring at 50 and 100% natural sediment mixtures, and at high (16 °C) temperature. ER was positively, but not significantly correlated with EHAA and chl a; a positive, significant correlation was detected between ER and sediment bacterial abundance. Overall, functional responses agreed with earlier compensatory intake models for deposit-feeders. However, the size and direction of these responses was temperature-sensitive, suggesting that these models need to be adjusted for changes in absorption rates. No effects of ambient temperature or food concentration on bacteriolytic rates were observed, possibly due to compensatory mechanisms or the presence of multiple bacteriolytic agents in gut fluids. Received: 28 June 1999 / Accepted: 14 March 2000     

The effect of light and heterotrophy on carotenoid concentrations in the Caribbean anemone<Emphasis Type="Italic"> Aiptasia pallida</Emphasis> (Verrill)

One of the consequences of ultraviolet radiation exposure in anthozoans possessing photosynthetic endosymbionts (i.e., zooxanthellae) is production of reactive oxygen species that can damage cellular components, especially lipids and photosynthetic membranes. It is well known that carotenoids are potent antioxidants that can mitigate oxygen radical damage, but the origin of these compounds in zooxanthellate anthozoans is obscured because they can potentially originate from endosymbionts, heterotrophic feeding by the host, or a combination thereof. We used Aiptasia pallida, a common Caribbean anemone, to investigate the effects of exogenous food sources, ultraviolet-A radiation (UVA, 320–400 nm), and photosynthetically active radiation (PAR, 400–700 nm) on carotenoid levels in zooxanthellate anthozoans. Anemones were exposed to one of three simulated light treatments in the laboratory for 38 days: PAR (60% below ambient)/UVA (similar to ambient), PAR/low UVA (42% below ambient), and low PAR (98.4% below ambient)/no UVA. In addition, anemones were either fed a carotenoid-rich diet of Artemia salina nauplii, or starved. Carotenoids identified in A. pallida included peridinin, diadinoxanthin, diatoxanthin and -carotene. While a diet of Ar. salina nauplii had no effect on the carotenoid composition of A. pallida, a two-way analysis of variance revealed that anemones exposed to ambient UVA levels had significantly greater diatoxanthin concentrations relative to the total xanthophyll pool [diato:(diato+diadino)] after 10 days of exposure. This difference among treatments was not present at 20 days, but reappeared as an effect due to starvation rather than UVA at days 30 and 38. These results suggest that carotenoids in A. pallida are not influenced by exogenous feeding and that photoprotective xanthophyll cycling is sensitive to stresses such as UVA and starvation.Communicated by P.W. Sammarco, Chauvin     

“Seed bank” of coastal planktonic diatoms in bottom sediments of Hiroshima Bay, Seto Inland Sea, Japan

Abundance and temporal distribution of viable (able to germinate) resting stage cells of planktonic diatoms in bottom sediments have been investigated almost monthly during 1989 to 1992 in Hiroshima Bay, western part of Seto Inland Sea, Japan. The abundance of viable resting stages in bottom sediments was enumerated with the extinction dilution method (most probable number method, MPN). In bottom sediments of Hiroshima Bay, dominantly distributed species and/ or genera of the diatom resting stages were Skeletonema costatum, Chaetoceros spp. and Thalassiosira spp. Viable resting stages of these diatoms were densely distributed on the orders of 10³ to 10⁶ (MPN g⁻¹ wet sediments), and persisted in bottom sediments throughout the investigation period. Conversely, vegetative cells of these diatoms fluctuated remarkably in the water column and disappeared sporadically. Survival of the resting stages in a collected sediment sample was also determined with the MPN method, at different storage temperatures (5, 10, 15, 20, 25 °C). The survival test demonstrated that the diatom resting stages could survive in the dark for several months or years in sediments. Resting stages survived longer at the lower storage temperature, and the order of longevity was consistent within three diatoms (Chaetoceros spp. > Thalassiosira spp. > S. costatum) at each storage temperature. The present study suggests that these diatom resting stages in the coastal bottom sediments could serve as a “seed bank”, analogous to those of terrestrial plants. The seed bank would ensure the survival of diatoms within highly fluctuating coastal environments, while it would also be the source of sporadic and autochthonous diatom blooms in coastal waters. Received: 29 November 1996 / Accepted: 16 December 1996     

Disease resistance: a benefit of sociality in the dampwood termite Zootermopsis angusticollis (Isoptera: Termopsidae)

The benefit of sociality in relation to disease susceptibility was studied in the dampwood termite Zootermopsis angusticollis. Although contact with high concentrations of fungal conidia is lethal, the survivorship of nymphs exposed to spore suspensions ranging from 6 × 10⁶ to 2 × 10⁸ spores/ml of the fungus Metarhizium anisopliae increased with group size. The survivorship (measured as LT₅₀) of isolated individuals ranged from 3.0 to 4.8 days, but infected nymphs living in groups of 10 and 25 individuals survived significantly longer (5.6–8.3 and 5.6–9.1 days, respectively). In most cases, there were no significant differences in the survival distributions of the 10- and 25-termite groups. When nymphs were infected with concentrations of 7 × 10¹–7 × 10⁴ spores/ml and allowed to interact with healthy nestmates, fungal infections were not contracted by the unexposed termites. Moreover, infected termites benefitted from social contact with unexposed nestmates: their survival rates were significantly higher than those of infected termites living with similarly infected nestmates. Allogrooming, which increased in frequency during and after exposure to conidia, appeared to remove potentially infectious spores from the cuticle, thus increasing termite survivorship. These results suggest that allogrooming plays a crucial role in the control of disease and its death hazard in termites. The infection-reducing advantage of group living may have been significant in the evolution of social behavior in the Isoptera. Received: 18 March 1998 / Accepted after revision: 31 May 1998     

Effect of nutrient enrichment in the field on the biomass, growth and calcification of the giant clam Tridacna maxima

Nutrients were added separately and combined to an initial concentration of 10 μM (ammonium) and/or 2 μM (phosphate) in a series of experiments carried out with the giant clam Tridacna maxima at 12 microatolls in One Tree Island lagoon, Great Barrier Reef, Australia (ENCORE Project). These nutrient concentrations remained for 2 to 3 h before returning to natural levels. The additions were made every low tide (twice per day) over 13 and 12 mo periods for the first and second phase of the experiment, respectively. The nutrients did not change the wet tissue weight of the clams, host C:N ratio, protein content of the mantle, calcification rates or growth rates. However, ammonium (N) enrichment alone significantly increased the total population density of the algal symbiont (Symbiodinium sp.: C = 3.6 · 10⁸ cell clam⁻¹, N = 6.6 · 10⁸ cell clam⁻¹, P = 5.7 · 10⁸ cell clam⁻¹, N + P = 5.7 · 10⁸ cell clam⁻¹; and C = 4.1 · 10⁸ cell clam⁻¹, N = 5.1 · 10⁸ cell clam⁻¹, P = 4.7 · 10⁸ cell clam⁻¹, N + P = 4.5 · 10⁸ cell clam⁻¹, at the end of the first and second phases of the experiment, respectively), although no differences in the mitotic index of these populations were detected. The total chlorophyll a (chl a) content per clam but not chlorophyll a per cell also increased with ammonium addition (C = 7.0 mg chl a clam⁻¹, N = 13.1 mg chl a clam⁻¹, P = 12.9 mg chl a clam⁻¹, N + P = 11.8 mg chl a clam⁻¹; and C = 8.8 mg chl a clam⁻¹, N = 12.8 mg chl a clam⁻¹; P = 11.2 mg chl a clam⁻¹, N + P = 11.3 mg chl a clam⁻¹, at the end of the first and second phases of the experiment, respectively). The response of clams to nutrient enrichment was quantitatively small, but indicated that small changes in inorganic nutrient levels affect the clam–zooxanthellae association. Received: 2 June 1997 / Accepted: 9 June 1997     

Do toxicity and accumulation of copper change during size reduction in the marine pennate diatom Haslea ostrearia?

 One of the best-known features of diatom biology is the reduction in mean cell size during vegetative multiplication by binary fission. We examined changes in copper toxicity and copper accumulation during cell-size reduction in Haslea ostrearia (Simonsen), a pennate diatom responsible for greening in oyster-ponds. We selected three strains with apical axes of different lengths: 40 μm (S40), 65 μm (S65) and 85 μm (S85). Each strain was grown separately in batch culture and exposed to a range of copper overloads (0 to 1.57 μM) that were added to the culture immediately after cell inoculation. Significant differences in sensitivity to copper were observed among the three strains. S85 exhibited highest sensitivity, followed by S40, while S65 displayed the highest tolerance. After 5 to 6 d exposure to 0.47 μM copper, chlorophyll a, carbohydrate, protein and lipid content per g dry weight had not changed in any of the three strains studied, except for a decrease of 16% in chlorophyll a in S85. At the end of the growth period with 0.47 μM copper, the amount of metal per unit surface area was similar for all strains, but the quantity of intracellular copper per g dry weight was lower in S65 cells than in S40 and S85 cells. Notable differences in the kinetics of both adsorbed and intracellular copper were observed between S40 and S85. Our results suggest that tolerance mechanisms may change during the vegetative life of H. ostrearia. The differential sensitivities of the strains suggest that copper pollution may alter the cell composition of natural populations of H. ostrearia by inducing selection for smaller cell size. Since auxosporulation results in the formation of larger cells with a higher sensitivity to copper, H. ostrearia could gradually disappear from copper-contaminated environments. In addition, by inducing smaller cell size, copper contamination would have an impact on filter-feeders such as oysters, whose diet is largely composed of diatoms. Received: 26 March 1999 / Accepted: 2 December 1999     

Importance of cysts in the population dynamics of the red tide flagellate Heterosigma akashiwo (Raphidophyceae)

To elucidate roles of cysts in occurrences of Heterosigma akashiwo blooms, cyst dynamics were studied in northern Hiroshima Bay, the Seto Inland Sea of Japan, where H. akashiwo regularly forms red tide in June. Monthly measurements of seasonal changes in the densities of vegetative cells of H. akashiwo and their germinable cysts in surface sediments (top 1-cm layer) were made for 2 years at three stations. Vegetative cells of H. akashiwo could be detected from April through December throughout the water column, and the existence of vegetative cells was confirmed in surface waters even in winter after incubation of sampled seawater in culture medium. Germinable cysts, enumerated by the extinction dilution method, existed in sediments in all seasons, even before and after the seasonal bloom. The effects of incubation temperature on the germination of natural cysts of H. akashiwo in sediments were examined. Germination was not observed at 5 °C, was low at 10 °C, while it increased at 15 °C, and maintained a high level to 25 °C. The bottom water temperature reached 15 °C (suitable for the germination of cysts) and the surface about 18 °C or more (suitable for the growth of vegetative cells) 2 to 3 weeks before the blooms. The dark survival of H. akashiwo cysts was tested, and it was found that the cysts were viable for at least 650 d at 11 °C, and for 165 d at 25 °C, indicating a significant role of cysts in the survival during winter and summer seasons. The cysts presumably also play an important role in seeding primary populations into water columns when the bottom water reaches a suitable temperature (around 15 °C); thereafter the populations develop with great annual regularity to bloom in June. These results suggest that initiation of H. akashiwo red tides in the Seto Inland Sea could be triggered by bottom water temperature. Received: 3 July 1998 / Accepted: 12 January 1999     

Light acclimation states of phytoplankton in the Southern Ocean, determined using photosynthetic pigment distribution

In high-latitude waters such as the Southern Ocean, the primary production of phytoplankton supports the ecosystem. To understand the photo-acclimation strategy of such phytoplankton within cold environments, the vertical distribution profile of photosynthetic pigments was analyzed in the Southern Ocean. Samples were taken along 110°E during the austral summer, and along 150°E and around the edge of the seasonal sea ice of the Antarctic Continent during the austral autumn. Pigment extraction methods were optimized for these samples. The standing crop of chlorophyll a was larger in the region along the edge of the seasonal sea ice than at sampling stations in open ocean areas. Chlorophyll concentration seemed to be dependent on the formation of thermo- and haloclines along the edge of the seasonal sea ice, but not in the open ocean where such clines are less pronounced. The marker pigments fucoxanthin and/or 19′-hexanoyloxyfucoxanthin were dominant at most sampling stations throughout the water column, while other marker pigments such as alloxanthin were quite low. This indicated that diatoms and/or haptophytes were the major phytoplankton in this area. Comparison of the relative ratio of fucoxanthin with that of 19′-hexanoyloxyfucoxanthin allowed some stations to be characterized as either diatom-dominant or haptophyte-dominant. The relative ratio of xanthophyll-cycle pigments (diadinoxanthin plus diatoxanthin) to chlorophyll a was high in surface waters and decreased gradually with depth. This suggests that near the ice edge during summer in the Southern Ocean, both diatoms and haptophytes acclimate to their light environments to protect their photosystems under high-light conditions.     

Interplay of host morphology and symbiont microhabitat in coral aggregations

The Belizean reef coral Agaricia tenuifolia Dana forms aggregations in which rows of thin, upright blades line up behind each other. On average, the spacing between blades increases with depth and hence with decreasing ambient irradiance. We designed and built a small, inexpensive light meter and used it to quantify the effect of branch spacing on light levels within colonies at varying distances from branch tips. Concurrently, we measured photosynthetic pigment concentrations and population densities of symbiotic dinoflagellates (zooxanthellae) extracted from coral branches of colonies with tight (≤3 cm) vs wide (≥6 cm) branch spacing, collected at 15 to 17 m and from colonies with tight branch spacing collected at 1 to 2 m. Light levels decreased significantly with tighter branch spacing and with distance from the branch tips. Total cellular pigment concentrations (chlorophylls a, c ₂ and peridinin) as well as chlorophyll a:c ₂ and chlorophyll a: peridinin ratios all increased significantly with distance from the branch tip, indicating very localized differences in photoacclimation within individual branches. Zooxanthellae from colonies with widely-spaced branches displayed significantly lower chlorophyll a:c ₂ and chlorophyll a:peridinin ratios, and were present at significantly higher population densities than those from colonies with tightly-spaced branches collected at the same depth (15 m). Tightly-spaced colonies collected from shallow environments (1 to 2 m) displayed pigment ratios similar to those from widely-spaced colonies from deeper water (15 m), but maintained zooxanthellae populations at levels similar to those in tightly-branched colonies from deeper water. Thus, variation in colony morphology (branch spacing and distance from branch tip) can affect symbiont physiology in a manner comparable to an increase of over 15 m of water depth. These results show that a host's morphology can strongly determine the microhabitat of its symbionts over very small spatial scales, and that zooxanthellae can in turn display steep gradients in concordance with these altered physical conditions. Received: 12 June 1997 / Accepted: 24 June 1997     

Nutrient stimulation of spore settlement in the kelpsPterygophora californica andMacrocystis pyrifera

FertilePterygophora californica Rupr. andMacrocystis pyrifera (L.) C.Ag. were collected in California, USA, from 1987 to 1989. Settlement activity of the spores was stimulated by nutrients, but this was not constant over time. Nutrients had no effect on the settlement ofP. californica spores between 2 and 18 h after sunrise, but settlement activity was stimulated by a nutrient mixture between 20 and 24 h after sunrise (14 to 18 h after release). Settlement activity inM. pyrifera spores was unaffected at 2 to 3 h after release, but settlement activity was significantly stimulated from 5 to 12 h after release. A variety of individual nutrients significantly stimulated settlement in 8 to 9 h oldM. pyrifera spores: ammonium, nitrate, glycine, phosphate, manganese-EDTA (18µM), borate, ferrous iron-EDTA, and ferric iron-EDTA. Spores also settled readily in unenriched artificial seawater, and nutrient-stimulated settlement rates were usually 150% of the unenriched control levels. Neither EDTA alone, cobalt-EDTA, nor manganese-EDTA (2µM) had significant effects onM. pyrifera spore settlement. The effects of time and of several individual nutrients on spore settlement activity are different from previously reported chemotactic effects of nutrients onP. californica andM. pyrifera spores. It is suggested that nutrient settlement-stimulation is mechanistically different from nutrient chemotaxis. However, like chemotaxis, settlement stimulation is probably an adaptation which increases the likelihood of spore settlement in microhabitats suitable for subsequent growth and reproduction of gametophytes.     

Cell density, chemical composition and toxicity of Chrysochromulina polylepis (Haptophyta) in relation to different N:P supply ratios

The influence of different N:P supply ratios on cell accumulation, chemical composition and toxicity of the marine haptophyte Chrysochromulina polylepis was examined in semi-continuous cultures. A non-axenic strain of C. polylepis was exposed to five different N:P supply ratios (N:P = 1:1, 4:1, 16:1, 80:1 and 160:1, by atoms), in order to create a range of N- and P-limited conditions. The toxicity per cell in C. polylepis was determined on four occasions at steady state cell density using the haemolytic activity of the cells expressed as saponin nanoequivalents. Haemolytic activity was demonstrated in all treatments, and increased in the algae when cell growth was nutrient limited (N:P = 1:1, 4:1, 80:1 and 160:1), compared to cells grown under non-limiting conditions (N:P = 16:1). This occurred regardless of the growth-limiting nutrient (N or P) and became more pronounced as nutrient limitation increased. In P-limited cultures the haemolytic activity per cell increased linearly with the cellular N:P ratio, whereas the N-limited cultures showed an opposite trend. The haemolytic activity per cell showed an inverse relationship with both cellular N and cellular P content. Cells limited by P showed a higher haemolytic activity than cells limited by N. The results suggest that toxicity in C. polylepis is strongly influenced by the physiological state of the algae. This may partially explain the large variability previously observed in the toxicity of C. polylepis blooms. The potential ecological significance of our findings is also discussed. Received: 18 November 1998 / Accepted: 5 July 1999     

Resource acquisition in two intertidal fucoid seaweeds, Fucus serratus and Himanthalia elongata : seasonal variation and effects of reproductive development

Seasonal variations and the effect of reproductive development on resource acquisition by two intertidal fucoid species, the iteroparous Fucus serratus L. and the semelparous Himanthalia elongata (L.) S. F. Gray were examined. The oxygen-exchange characteristics of vegetative apical tissue of both non-fertile and fertile plants and receptacle tissue were compared at monthly intervals throughout reproductive development. Respiratory rates in non-fertile F. serratus varied seasonally between 1.5 and 8.0 μmol g⁻¹ fresh wt h⁻¹; in fertile plants the receptacle had a significantly lower respiratory rate than the vegetative tissue. The respiratory rate of the vegetative button of fertile H. elongata displayed less seasonal variation and was lower than that of the receptacle, which varied from a maximum of 9.5 μmol g⁻¹ fresh wt h⁻¹ at receptacle initiation in October to a minimum of 2.0 μmol g⁻¹ fresh wt h⁻¹ in February. The maximum photosynthetic rate (P _max) of non-fertile plants of both species did not vary in a distinct seasonal manner (∼60 μmol g⁻¹ fresh wt h⁻¹ for F. serratus and ∼12 μmol g⁻¹ fresh wt h⁻¹ for H. elongata). In fertile plants, the P _max of the receptacle tissue was (∼50% lower in F. serratus, and at its peak three times higher in H. elongata, than that of vegetative tissue. The stable carbon-isotope ratio (δ¹³C) did not differ between different tissue types in F. serratus, but values did vary seasonally, being less negative in the summer than in the winter (−13.5‰ compared to −18‰). The receptacle tissue of H. elongata also displayed a distinct seasonal variation in δ¹³C values (−12‰ in summer, −16‰ in winter), whilst the δ¹³C of the vegetative button did not vary seasonally. The rate of uptake of inorganic nitrogen by the vegetative thallus was lower in H. elongata than in F. serratus. The receptacle tissue of F. serratus had lower uptake rates than the vegetative tissue, whilst the uptake rate by H. elongata receptacle tissue was higher than that of the vegetative button. Received: 14 March 1997 / Accepted: 22 April 1997     

UV- and salinity-induced oxidative effects in the marine diatom <Emphasis Type="Italic">Cylindrotheca closterium</Emphasis> during simulated emersion

The diatom Cylindrotheca closterium was exposed to transient light- and osmotic conditions as occur during its tidal emersion. The objective was to analyze how this simulated emersion contributes to the production of active oxygen species (AOS) and via this, to oxidative cell damage. Light- and salinity conditions were varied in factorial combination: low light (no UVB) or high light (unweighted UVB-dose rates of respectively 0.01; 0.07; 0.24; 1.03 W m⁻²) at normal (30 psu) or high salinity (60 psu). UVB (0.01–0.24 W m⁻²) and high salinity had a significant, negative effect on the photosynthetic efficiencies ΔF/F _m’ (steady-state quantum yield) and F _v/F _m (maximum yield). UVB at 1.03 W m⁻² (15 kJ m⁻² d⁻¹) almost arrested electron transport. At ecologically relevant UVB levels, i.e. below 0.24 W m⁻² (≈3.4 kJ m⁻² d⁻¹) with UVB:PAR<0.4:100 (PAR photosynthetically active radiation) only dynamic photoinhibition was observed (protection via heat dissipation). Non-photochemical quenching was positively correlated with the de-epoxidation of diadinoxanthin (DD) to diatoxanthin (DT). A decreasing ratio DT/(DD+DT) after 4 h of UVB at >0.07 W m⁻² and at 60 psu indicated a reversal of the diatom xanthophyll cycle (diminished photoprotection) which may be caused by an enhanced AOS production. Oxidative stress and -damage to C. closterium cells were assessed applying fluorescent indicator dyes, via confocal microscopy and quantitative image analysis. AOS production rates (cellular DCF fluorescence) were stimulated by UV, and were ~50% higher at 60 psu. AOS production decreased with an increasing pre-exposure (0–4 h) to normal UVB (0.24 W m⁻²), which indicated a stimulation of the antioxidative defence. Non-protein thiols (indicator CMF) and glutathione pools (HPLC-analyzed) decreased with UVB-dose rates (0.01–0.24 W m⁻²), most likely due to AOS-mediated thiol oxidation. Hypersalinity (60 psu) and UVB (0.01–0.24 W m⁻²) caused membrane depolarization (dye DIBAC₄(3)) and phospholipid hydrolysis (phospholipase A₂ dye: bis-BODIPY FL-C₁₁-PC). AOS production may have diminished the membrane polarity, and peroxidized the membrane lipids (HPLC-analyzed malondialdehyde) which enhanced PLA₂ activity. The dyes indicated an increased oxidative (lipid) damage at a 15% inhibition of photosynthesis in this diatom, at UVB levels and salinities that can be expected in situ during its periodic tidal emersion.     

The use of nondestructive methods to assess a physiological status and conservation perspectives of <Emphasis Type="Italic">Eryngium maritimum</Emphasis> L.

Eryngium maritimum is a perennial species growing exclusively in a coastal littoral zone both on sand dunes and shingle beach and indicated as declining in Northern Europe. The objective of the present study was to prove the use of nondestructive physiological measurement methods to access physiological status of endangered plants, using E. maritimum as a model species. Plants from two Latvian sites were studied in comparison with other populations in Northern Europe (Estonia, Lithuania, Poland, United Kingdom) to find out if local differences in environmental factors affect vitality of E. maritimum individuals. Noninvasive chlorophyll analysis and chlorophyll a fluorescence measurements were used as indicators of plant physiological status through characterization of various aspects of photochemistry of photosystem II activity. Dynamics and morphology of individuals were investigated for evaluation of clonal growth potential of E. maritimum in natural conditions. Highly fluctuating trend of dynamics of individuals (within 40 groups at two Latvian sites) was established for E. maritimum in natural conditions over a five-year period. Disturbance of individuals lead to formation of new shoots from nodal root meristems. An exponential regression between fluorescence parameters F_V/F_M and Performance Index and summary monthly precipitation was found indicating that E. maritimum plants had significant tolerance to water shortage together with susceptibility to increased precipitation. Negative correlation between leaf chlorophyll content and more northern localization of individuals was found (r = 0.95 and r = 0.94, for generative and vegetative shoots, respectively). Northern populations of E. maritimum are endangered by high precipitation in conditions of low air temperature, leading to decrease of photosynthetic productivity and overall physiological status. High developmental plasticity at the root level leads to clonal growth and an efficient survival and relatively long life span of E. maritimum individuals. Chlorophyll a fluorescence is a useful method to search for the effect of suboptimal conditions on physiological status of endangered plant species without elimination and disturbance of individuals.