A comparative analysis of the photobiology of zooxanthellae and zoochlorellae symbiotic with the temperate clonal anemone Anthopleura elegantissima (Brandt)

The temperate anemone Anthopleura elegantissima hosts two phylogenetically different symbiotic microalgae, a dinoflagellate Symbiodinium (zooxanthellae, ZX) and a chlorophyte (zoochlorellae, ZC), throughout certain regions of its latitudinal range. Because of the broad intertidal and geographic range of this anemone, we examined the role of irradiance to ascertain which specific symbiotic parameters are affected and whether light intensity governs the observed distributions of natural populations of ZX and ZC. Irradiance appears to be a key factor in regulating both the photophysiology and metabolism of this alga-cnidarian association. Regardless of light intensity, algal densities remained stable for anemones harboring ZX or ZC, whereas the mitotic indices of ZX and ZC both varied directly with light intensity. The chlorophyll content of ZX remained fairly constant regardless of irradiance; in contrast, ZC chlorophyll content was inversely proportional to light intensity. Regardless of irradiance, the carotenoid content of both symbionts was constant; however, ZX carotenoid levels were higher than those of ZC. Net photosynthesis was directly related to light intensity for both algal symbionts and ZX photosynthetic rates were consistently higher than those of ZC. Similarly, the potential carbon contribution of ZX and ZC to animal respiration (CZAR) displayed a direct relationship with light intensity, peaking at 800 µmol·m^-2·s^-1, then subsequently declined. Lower ZX growth rates, coupled with higher photosynthetic rates and higher CZAR estimates, compared to ZC, suggest that the ZX should be the dominant symbiont as light intensity increases; this may explain the high densities of anemones in the field containing ZX where the levels of irradiance are naturally high. These results support the interpretation that irradiance is a significant environmental parameter that dictates the microhabitat and latitudinal distribution of the two symbiotic algal taxa. This is the second in a series of papers examining the physical parameters that influence the distribution of ZX- and ZC-bearing A. elegantissima.     

A comparative analysis of the photobiology of zooxanthellae and zoochlorellae symbiotic with the temperate clonal anemone <Emphasis Type="Italic">Anthopleura elegantissima</Emphasis> (Brandt). III. Seasonal effects of natural light and temperature on photosynthesis and respiration

The sea anemone Anthopleura elegantissima hosts two phylogenetically different symbiotic microalgae, a dinoflagellate Symbiodinium (zooxanthellae, ZX) and a chlorophyte (zoochlorellae, ZC). The photosynthetic productivity (P), respiration (R), and contribution of algal carbon translocated to the host (CZAR) in response to a year’s seasonal ambient changes of natural light and temperature are documented for both ZX- and ZC-bearing anemones. Light and temperature both affect photosynthesis, respiration, and CZAR, as well as various algal parameters; while there are evident seasonal differences, for the most part the relative effects on P, R, and CZAR by the two environmental variables cannot be determined. Net photosynthesis (P_n) of both ZX and ZC was significantly higher during spring and summer. During these seasons, the P_n of ZX was always greater than that of ZC. Regardless of algal symbiont, anemone respiration (R) was significantly higher during the spring and summer. The annual net carbon fixation rate of anemones with ZX and ZC was 325 and 276 mg C anemone⁻¹ year⁻¹, respectively, which translates to annual net community productivity rates of 92 and 60 g C m⁻¹ year⁻¹ for anemones with ZX or ZC, respectively. CZAR did not show a clear relationship with season; however the CZAR for ZX was always significantly greater than for ZC. Lower ZX growth rates, coupled with higher photosynthetic rates and higher CZAR estimates, compared to ZC, suggest that if A. elegantissima is simply carbon limited, ZX-bearing anemones should be the dominant symbiont in the field. However ZC-bearing anemones persist in low light and reduced temperature microhabitats, therefore more than the translocation of carbon from ZC must be involved. Given that global climate change will increase water temperatures, the potential for latitudinal range shifts of both ZC and ZX (S. californium and muscatinei) might be used as biological indicators of thermal shifts in the littoral zone of the Pacific Northwest.     

The effects of symbiotic state on heterotrophic feeding in the temperate sea anemone <Emphasis Type="Italic">Anthopleura elegantissima</Emphasis>

The temperate sea anemone Anthopleura elegantissima is facultatively symbiotic with unicellular algae. Symbiotic A. elegantissima can supplement heterotrophic feeding with excess photosynthate from their algal partners, while asymbiotic individuals must rely solely on heterotrophy. A. elegantissima individuals were collected from Swirl Rocks, Washington (48°25′6″ N, 122°50′58″ W) in July 2010, and prey capture and feeding characteristics were measured to determine whether asymbiotic individuals are more efficient predators. Feeding abilities were then measured again after a 3-week exposure to full sunlight or shaded conditions. Freshly collected asymbiotic anemones had larger nematocysts, but symbiotic individuals showed greater nematocyte sensitivity. Sunlight enhanced digestion and reduced cnida density in all anemones regardless of symbiotic state. Results suggest that the phototropic potential of A. elegantissima, as influenced by symbiotic condition, has little effect on heterotrophic capacity. The anemones appear to maximize heterotrophic energy input independent of the presence or identity of their algal symbionts.     

Effects of starvation,and light and dark on the energy metabolism of symbiotic and aposymbiotic sea anemones,Anthopleura elegantissima

Rates of oxygen and carbon-dioxide exhange were measured in symbiotic and aposymbiotic specimens of the sea anemone Anthopleura elegantissima while fed and starved under light or dark conditions. Respiratory quotients indicated that fed anemones switched from a carbohydrate to a fat catabolism when starved, with the exception that symbiotic individuals starved in the light showed a pronounced carbohydrate catabolism for over 1 month. The source of the carbohydrate was probably photosynthate translocated by the dinoflagellate Symbiodinium (=Gymnodinium) microadriaticum (Freudenthal) living in the anemones' tissues. The starved symbiotic anemones maintained in the light had lipid levels not significantly different from fed controls and 44 to 61% higher than starved aposymbiotic anemones after 1 month. Thus, the quality and quantity of the metabolic flux from the symbionts to the sea anemone were sufficient to conserve the host's lipid reserves.     

Cellular and ultrastructural changes in the endoderm of the temperate sea anemone Anemonia viridis as a result of increased temperature

Exposure of the temperate sea anemone Anemonia viridis Forskål to increased seawater temperature (from 16 to 26°C) reduced the lysosomal latency of coelenterate tissues. Lysosomes in the mesenterial filaments of anemones were destabilised by increased temperature, with greater destabilisation in heat-shocked symbiotic anemones than in heat-shocked aposymbiotic anemones in the early stages of the experiment. Lysosomal enzyme activity in zooxanthellae from heat-shocked symbiotic anemones was associated with the algal membranes and the cytoplasm of degenerate algal cells. While the relationship between host coelenterate and symbiotic alga may confer many benefits under normal conditions, comparison of the responses of symbiotic and aposymbiotic anemones to heat shock suggests that there may be disadvantages for symbiotic anemones under stress.     

Effect of feeding regime and irradiance on the photophysiology of the symbiotic sea anemone Aiptasia pulchella

To determine how the animal and algal components of the symbiotic sea anemone Aiptasia pulchella respond to changes in food availability and culture irradiance, sea anemones from a single clone were maintained at four irradiance levels (320, 185, 115, and 45 E m^-2 s^-1) and either starved or fed for 5 wk. Changes in protein biomass of sea anemones maintained under these conditions were not related to the productivity of zooxanthellae, since the protein biomass of fed A. pulchella decreased with increase in irradiance and there was no difference in protein biomass among starved sea anemones at the four irradiance levels. Except for the starved high-light sea anemones, the density of symbiotic zooxanthellae was independent of culture irradiance within both starved and fed. A. pulchella. Starved sea anemones contained over twice the density of zooxanthellae as fed sea anemones. Within both starved and fed individuals, chlorophyll per zooxanthella increased with decreasing culture irradiance while algal size remained constant (in fed sea anemones) at about 8.80 m diameter. Chlorophyll a: c ₂ ratios of zooxanthellae increased with decreasing culture irradiance in zooxanthellae from starved sea anemones but remained constant in zooxanthellae from fed sea anemones. As estimated from mitotic index data, the in situ growth rates of zooxanthellae averaged 0.007 d^-1 and did not vary with irradiance or feeding regime. Photosynthesis-irradiance (P-I) responses of fed A. pulchella indicated an increase in photosynthetic efficiency with decreasing culture irradiance. But there was no consistent pattern in photosynthetic capacity with culture irradiance. Respiration rates of fed sea anemones also did not vary in relation to culture irradiance. The parameter I _k , defined as the irradiance at which light-saturated rates of photosynthesis are first attained, was the only parameter from the P-I curves which increased linearly with increasing culture irradiance. The daily ratio of net photosynthesis to respiration for A. pulchella ranged from 1.6 to 2.8 for sea anemones maintained at the three higher irradiances, but was negative for those maintained at 45 E m^-2 s^-1. Since the final protein biomass was greatest for sea anemones maintained at the lowest irradiance, these results indicate that sea anemone growth cannot be directly related to productivity of zooxanthellae in this symbiotic association.     

Physiological energetics of the intertidal sea anemone Anthopleura elegantissima

Energy budgets were calculated for individuals of the sea anemone Anthopleura elegantissima (Brandt), collected in 1981 and 1982 from Bodega Harbor, California, USA. Rates of ammonium excretion were measured in high-and low-intertidal, symbiotic and aposymbiotic sea anemones within 24 h of collection. Among symbiotic and aposymbiotic individuals, no differences in excretion rate were found on the basis of intertidal height. However, rates of ammonium excretion in aposymbiotic anemones (2.14 mol NH ₊ ⁴ g^-1 h^-1) were significantly higher than in symbiotic ones (0.288 mol NH ₊ ⁴ g^-1 h^-1). Rates of excretion were used with estimated rates of oxygen uptake to calculate nitrogen quotients (NQ). NQ and RQ values from the literature were used to calculate an oxyenthalpic equivalent [501 kJ (mol O₂)^-1 for R+U], and mass proportions of protein (54%), carbohydrate (44%) and lipid (2%) catabolized during routine metabolism in this species 24 h after feeding. Integrated energy budgets of these experimental anemones were calculated from data on ingestion, absorption and growth, and estimates of translocated energy from the symbiotic algae. Contribution of zooxanthellae to animal respiration based on translocation=90% and RQ=0.97 are 41 and 79% in high-and low-intertidal anemones, respectively. Calculated scope for growth is greater than directly measured growth in both high-and low-intertidal individuals. The deficit, estimated as 30% of assimilated energy in high-intertidal anemones, is attributed to unmeasured costs (specific dynamic effect) or production (mucus). Low-intertidal anemones lost mass during the experiment, implying that the magnitude of the deficit was greater in these anemones than in upper intertidal individuals. Anemones from both shore levels lost zooxanthellae during the experiment, which contributed to energy loss since the contribution of the zooxanthellae is greater in low-intertidal anemones. Scope for growth is preserved in high-intertidal anemones (29% of assimilated energy) because metabolic demands are lower due to aerial exposure, and prey capture rate is higher compared to lowshore anemones. Although possibly underestimated, lower scope for growth in low-shore anemones may result from continuous feeding and digestion processes that are less efficient than those of periodically feeding high-intertidal anemones.     

Oxygen as a limiting factor in phototaxis and in intraclonal spacing of the sea anemone Anthopleura elegantissima

Phototaxis in Anthopleura elegantissima, a sea anemone symbiotic with zooxanthellae, was investigated with special reference to oxygen as a possible controlling factor. Under high oxygen concentrations in seawater, movement towards light was not observed for symbiotic anamones as it was under normal oxygen concentrations. Both aposymbiotic and symbiotic anemones demonstrated movement towards high oxygen concentrations in seawater. Oxygen is, therefore, implicated as a controlling factor in phototaxis. Under laboratory conditions, increased intraclonal spacing occurred with low oxygen concentrations in seawater. In the field, individuals in symbiotic clones were spaced significantly closer than in aposymbiotic clones. Since intraclonal spacing is controlled by oxygen in the laboratory, spacing may also be affected in the field by oxygen; symbiotic clones may be spaced closer because they have better oxygen availability than do aposymbiotic clones.     

Physiological energetics of the intertidal sea anemone Anthopleura elegantissima

High-intertidal (H) individuals of the sea anemone Anthopleura elegantissima (Brandt) are exposed aerially up to 18 h each day, unlike low-intertidal (L) individuals which may be continuously immersed over many days. Thus, H anemones experience shorter feeding periods compared to L anemones. From 1980 to 1982, H and L anemones were observed and collected at the mouth of Bodega Harbor in North Central California (USA) to determine whether any physiological adaptations mitigate the energetic effects of reduced feeding time in H anemones. Weight of prey in coelenterons of H anemones was three times more than that of L anemones following a single immersion period. This difference is not due to slower digestion rates in H anemones. Prey residence time in coelenterons (4h) was equivalent in both groups. Different prey weights imply that ingestion rates were greater in H individuals. However, all anemones had similar weight-specific feeding-surface areas. Different prey-capture rates result from increased receptivity to prey in H anemones, rather than from increases in feeding surface. Absorption efficiency was inversely related to ration size in anemones from both shore positions. H individuals absorbed food more efficiently than L individuals fed equivalent rations. Ration, not exposure conditions, affected absorption efficiency. Daily growth rates were 1.5 to 1.8% and 1.2 to 1.4% of dry body weight in H and L anemones fed large rations (4.0 to 5.6% of dry body weight), respectively. H anemones fed smaller daily rations, approximating amounts of zooplankton captured naturally (1% of anemone dry weight), had higher growth rates and growth efficiencies than L anemones, which lost mass. Higher growth rates in H anemones, which are supported by higher prey-capture rates, result in attainment of minimum body size for reproduction in a relatively short period of time despite reduction in time available for feeding, thus improving relative fitness of these anemones in the upper intertidal zone.     

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     

Feeding behavior and acquisition of zooxanthellae by planula larvae of the sea anemone Anthopleura elegantissima

Symbiotic associations between cnidarians and photosynthetic dinoflagellates (i.e., zooxanthellae) are common in the marine environment. Many symbiotic cnidarians produce offspring that are initially nonsymbiotic. These new hosts must acquire symbiotic algae from environmental sources. We examined zooxanthella acquisition by laboratory-reared planula larvae of the temperate sea anemone Anthopleura elegantissima. Larvae ingested zooxanthellae while they were feeding. However, the signal that prompted larval feeding behavior did not originate from the symbiotic algae; the addition of algal cells to larval cultures never elicited a feeding response. In contrast, the addition of macerated animal tissue from several sources invariably generated a strong feeding response, which resulted in the larvae indiscriminately ingesting any particulate matter that was present, including zooxanthellae or other unicellular algae. Ingested zooxanthellae were incorporated into endodermal cells, where they remained undigested, while all other ingested material was digested or expelled within 24 h. Our results provide evidence that one source of zooxanthellae likely to serve as a route of infection in the natural environment is zooxanthella-laden mucus egested by anemones. This egested material fulfilled both of the criteria necessary for successful infection: it prompted larvae to begin feeding and provided an abundant supply of zooxanthellae that were ingested and taken up into endodermal cells of the new host.     

Viable algae released by the seastar <Emphasis Type="Italic">Dermasterias</Emphasis><Emphasis Type="Italic">imbricata</Emphasis> feeding on the symbiotic sea anemone <Emphasis Type="Italic">Anthopleura elegantissima</Emphasis>

Echinoderms are major predators of anemones in temperate ecosystems. The fate of two algae, zooxanthellae and zoochlorellae, after their host anemone (Anthopleura elegantissima Brandt) was consumed by the leather star Dermasterias imbricata Grube was determined in experiments conducted in July and August 2004. Productivity, photosynthetic pigments, and mitotic index (percent of cells dividing) were used as indicators of algal health; algae released after leather stars consumed their host were compared with algae freshly isolated from anemones. Two types of waste products contained algae: pellets resulting from extraoral digestion, and feces. Zooxanthellae and zoochlorellae isolated from these waste products were photosynthetic, although to different extents. For algae from feces and pellets, light-saturated photosynthetic rates (P _max) were 85 and 13%, respectively, of P _max of freshly isolated zooxanthellae; and were 20 and 46%, respectively, for zoochlorellae. The photosynthetic pigments and mitotic index (percent of dividing cells) were not altered by the feeding activities of the leather star. These results show that algae released by seastar predation on their hosts remain viable, and are hence available for establishing symbioses in A. elegantissima and other potential hosts.     

Production and biomass accumulation of periphytic diatoms growing on glass slides during a 1-year cycle in a subtropical estuarine environment (Bay of Paranaguá, southern Brazil)

Production rates, chlorophyll concentrations and general composition of periphytic diatom communities growing on glass slides were studied in relation to environmental parameters during one seasonal cycle in the Bay of Paranaguá, southern Brazil. Slides were routinely submersed at 1, 2 and 3 m depth and recovered weekly for microscopic examinations, analyses of chlorophyll, cell counts and in situ photosynthetic incubations using the Winkler titration method. Water samples were also collected at surface and bottom layers for determinations of temperature, salinity, nutrients and chlorophyll in the water. The periphytic community was mainly formed by epipelic and epipsammic species, dominated by Navicula phyllepta, Cylindrotheca closterium, Navicula spp. and Amphora sp. Weekly chlorophyll a and cell accumulations on slides varied from <1–32 mg m⁻² and up to 31 × 10⁸cells m⁻², respectively. Photosynthetic rates varied from <1 to 35 mg oxygen mg chlorophyll a ⁻¹ h⁻¹, with higher values in summer. Daily production varied from 5 to 3,600 mg oxygen m⁻² day⁻¹ (<0.01–1.4 g carbon m⁻² day⁻¹). Multiple regression analysis revealed that vertical differences in light conditions and grazing pressure jointly affected the influence of temperature on the seasonal patterns of cell densities and chlorophyll concentrations according to depth. Received: 27 April 2000 / Accepted: 16 August 2000     

Host feeding and nutrient sufficiency for zooxanthellae in the sea anemone Aiptasia pallida

Nutrient sufficiency of zooxanthellae in the sea anemone Aiptasia pallida cultured in low nutrient seawater depends on the availability of particulate food to the host. Zooxanthellae in anemones unfed for 20 to 30 d exhibited the following characteristics of nutrient deficiency: cell division rates decreased; chlorophyll a content gradually decreased from 2 to <1 pg cell^–1; and C:N ratios increased from 7.5 to 16. Over a 3-mo period, algal populations in unfed anemones gradually decreased, indicating that zooxanthellae were lost faster than they were replaced by division. The mitotic index of zooxanthellae in unfed anemones was stimulated either by feeding the host or by the addition of inorganic N and P to the medium. Whether algae are nutrient-limited in hosts under field conditions has not been examined fully; however, C:N ratios in zooxanthellae from field-collected hosts are slightly higher (9.4 vs 7.5) than in hosts fed to repletion in laboratory cultures. This observation might indicate N limitation in the field.     

Distribution of lipids between the zooxanthellae and animal compartment in the symbiotic sea anemoneAnemonia viridis: Wax esters,triglycerides and fatty acids

The temperate sea anemoneAnemonia viridis (Forskäl) contained about 11% lipid on a dry weight basis when maintained at light levels of about 10µE m^–2 s^–1 and a temperature of 10°C. Aposymbiotic forms of the anemone had similar lipid levels. These values are very low compared with tropical symbiotic Anthozoa in which lipid levels constitute up to 50% of dry weight. In symbioticA. viridis, <6% of total lipid consisted of the storage lipids, wax esters and triglycerides. Most of the triglyceride was stored in the animal tissues rather than the zooxanthellae. Zooxanthellae contained only small amounts of wax esters. An analysis was made of the wax ester, triglyceride and fatty acid composition of symbiotic anemones, isolated zooxanthellae and aposymbiotic anemones. Wax ester composition was similar in symbiotic and aposymbiotic forms. However, triglyceride composition differed. In particular trimyristin (C42) was found only within the symbiotic association. Fatty acids showed a high degree of unsaturation, and acids with both even and odd numbers of carbon atoms were found. The most abundant fatty acid was 16:0 in all samples, except for the total lipids from zooxanthellae in which the major fatty acid wastrans-18:1.     

Microsensor studies of photosynthesis and respiration in the symbiotic foraminifer Orbulina universa

Oxygen and pH microelectrodes were used to investigate the microenvironment of the planktonic foraminifer Orbulina universa and its dinoflagellate endosymbionts. A diffusive boundary layer surrounds the foraminiferal shell and limits the O₂ and proton transport from the shell to the ambient seawater and vice versa. Due to symbiont photosynthesis, high O₂ concentrations of up to 206% air saturation and a pH of up to 8.8, i.e. 0.5 pH units above ambient seawater, were measured at the shell surface of the foraminifer at saturating irradiances. The respiration of the host–symbiont system in darkness decreased the O₂ concentration at the shell surface to <70% of the oxygen content in the surrounding air-saturated water. The pH at the shell surface dropped to 7.9 in darkness. We measured a mean gross photosynthetic rate of 8.5 ± 4.0 nmol O₂ h⁻¹ foraminifer⁻¹. The net photosynthesis averaged 5.3 ± 2.7 nmol O₂ h⁻¹. In the light, the calculated respiration rates reached 3.9 ± 1.9 nmol O₂h⁻¹, whereas the dark respiration rates were significantly lower (1.7 ± 0.7 nmol O₂ h⁻¹). Experimental light–dark cycles demonstrated a very dynamic response of the symbionts to changing light conditions. Gross photosynthesis versus scalar irradiance curves (P vs E _o curves) showed light saturation irradiances (E _k) of 75 and 137 μmol photons m⁻² s⁻¹ in two O. universa specimens, respectively. No inhibition of photosynthesis was observed at irradiance levels up to 700 μmol photons m⁻² s⁻¹. The light compensation point of the symbiotic association was 50 μmol photons m⁻² s⁻¹. Radial profile measurements of scalar irradiance (E _o) inside the foraminifera showed a slight increase at the shell surface up to 105% of the incident irradiance (E _d). Received: 26 January 1998 / Accepted: 11 April 1998     

Coexistence of nine anemonefish species: differential host and habitat utilization, size and recruitment

The region of Madang, Papua New Guinea, has the highest reported species diversity of both anemonefishes (nine species) and their host anemones (ten species). To determine which factors may allow so many anemonefish species to coexist at this location, we studied their patterns of distribution, abundance, and recruitment. Population surveys at three replicate reef sites within four zones situated at varying distances from the mainland (nearshore, mid-lagoon, outer barrier, and offshore) indicated that each species of host anemone and anemonefish lived within a particular range of zones. Each species of anemonefish lived primarily with one species of host. Anemonefish species that lived with the same host species usually had different distribution patterns among zones (e.g., Amphiprion percula occupied Heteractis magnifica in nearshore zones, while A. perideraion occupied H. magnifica in offshore zones). Monitoring of natural populations showed that there were few changes (losses or recruitment) in the number or species of fishes associated with each individual anemone over periods ranging from 3 to 9 months. Recruitment was monitored on anemones with and without residents (resident fishes were removed) within each of three zones (nearshore, mid-lagoon, outer barrier). Significantly more anemonefishes recruited to anemones without resident fishes than to anemones with resident fishes. Each anemonefish species recruited to particular host species and zones. The distribution and abundance of the recruits of each fish species among zones were positively correlated with the distribution and abundance of resident fishes in the benthic habitat. This suggests that the spatial patterns of recruitment among zones strongly determined the distribution and abundance patterns of the benthic populations, and they were not the result of post-recruitment mortality or movement. Coexistence of the nine anemonefish species on the limited anemone resource was considered possible because of niche differentiation (i.e., differences in host and habitat utilization among zones), and the ability of two small species (i.e., Amphiprion sandaracinos and A. leucokranos) to cohabit individual anemones with other anemonefish species. Received: 29 July 1999 / Accepted: 1 September 2000     

Genetic relationships within and between clonal and solitary forms of the sea anemone Anthopleura elegantissima revisited: evidence for the existence of two species

Along the temperate Pacific coast of North America, the actiniarian sea anemone Anthopleura elegantissima exhibits two discrete life-history phenotypes. Although both forms sexually produce planula larvae, the clonal morph can also asexually propagate by fission, whereas the solitary morph does not. Whether the two forms constitute one or two species has long been contested. Hand originally designated the two forms as conspecifics, whereas Francis – on the basis of differences in microhabitat, biogeographic range and phenotypic frequencies – argued that the two forms constituted a sibling-species pair. From the results of an electrophoretic survey in which they pooled allelic frequencies across several geographic locations, Smith and Potts subsequently argued that the two forms were not genetically differentiated, and therefore represented a single species. We re-examined the relationship between the forms electrophoretically, substantially extending the geographic range and doubling the sample sizes beyond those used by Smith and Potts, and not pooling allelic frequencies in our analyses. Our analysis of patterns of genetic variation at ten highly polymorphic allozyme loci shows that although no fixed genetic differences distinguish the two forms, there are significant differences in allele frequencies between clonal and solitary A.␣elegantissima at every site we sampled throughout their range of sympatry (over 1000 km); within each form, however, there is little detectable genetic differentiation among populations. We therefore conclude that the two forms represent recently reproductively isolated taxa, and propose that the clonal form retain the binomial A. elegantissima (Brandt, 1835), whereas the solitary form be described and named a new species, Anthopleura sp. Received: 28 August 1996 / Accepted: 25 September 1996     

Symbiotic anemones can grow when starved: nitrogen budget for Anemonia viridis in ammonium-supplemented seawater

The ability of endosymbioses between anthozoans and dinoflagellate algae (zooxanthellae) to retain excretory nitrogen and take up ammonium from seawater has been well documented. However, the quantitative importance of these processes to the nitrogen budget of such symbioses is poorly understood. When starved symbiotic Anemonia viridis were incubated in a flow-through system in seawater supplemented with 20 μM ammonium for 91 d under a light regime of 12 h light at 150 μmol photons m⁻² s⁻¹ and 12 h darkness, they showed a mean net growth of 0.197% of their initial weight per day. Control anemones in unsupplemented seawater with an ammonium concentration of <1 μM lost weight by a mean of 0.263% of their initial weight per day. Attempts to construct a nitrogen budget showed that, over a 14 d period, ≃40% of the ammonium taken up could be accounted for by growth of zooxanthellae. It was assumed that the remainder was translocated from zooxanthellae to host. However, since the budget does not balance, only 60% of the growth of host tissue was accounted for by this translocation. The value for host excretory nitrogen which was recycled to the symbionts equalled that taken in by ammonium uptake from the supplemented seawater, indicating the importance of nitrogen retention to the symbiotic association. Received: 23 December 1997 / Accepted: 12 September 1998     

Cell cycle of symbiotic dinoflagellates: variation in G1 phase-duration with anemone nutritional status and macronutrient supply in the Aiptasia pulchella–Symbiodinium pulchrorum symbiosis

The metabolite exchange in alga–invertebrate symbioses has been the subject of extensive research. A central question is how the biomass of the algal endosymbionts is maintained within defined limits under a given set of environmental conditions despite their tremendous growth potential. Whether algal growth is actively regulated by the animal cells is still an open question. We experimentally evaluated the effect of inorganic nutrient supply and host-animal nutritional status on the biomass composition, growth and cell-cycle kinetics of the endosymbiotic dinoflagellate Symbiodinium pulchrorum (Trench) in the sea anemone Aiptasia pulchella. Dinoflagellates in anemones starved for 14?d exhibited lower growth rates, chlorophyll content and higher C:N ratios than in anemones fed Artemia sp. (San Francisco brand #65034) nauplii every 2 d, indicating N-limitation of the algae during starvation of the host animal. Manipulation of the dissolved inorganic nutrient supply through ammonium and phosphate additions induced a rapid recovery (half time, t _½～ 2?d) in the C:N ratio of the dinoflagellate cells to levels characteristic of N-sufficient cells. The mitotic index and population growth rate of the dinoflagellate symbionts subjected to this enrichment did not recover to the levels exhibited in fed associations. Flow cytometric analysis of dinoflagellate cell size and DNA content revealed that the duration of the G₁ phase (first peak of DNA content: 70 to 100 relative fluorescence units, rfu) of their cell cycle lengthened dramatically in the symbiotic state, and that the majority of algal biomass increase occurred during this phase. Covariate analysis of dinoflagellate cell size and DNA-content distributions indicated that the symbiotic state is associated with a nutrient-independent constraint on cell progression from G₁ through the S phase (intermediate DNA content: 101 to 139?rfu). This analysis suggests that the host-cell environment may set the upper limit on the rate of dinoflagellate cell-cycle progression and thereby coordinate the relative growth rates of the autotrophic and heterotrophic partners in this symbiotic association.