New examples of holococcolith–heterococcolith combination coccospheres and their implications for coccolithophorid biology

Recently discovered coccospheres with combinations of coccoliths normally considered to belong to different taxa are presented here. By analogy with other coccolithophorids especially Coccolithus pelagicus we can hypothesize that these associations probably represent moments of phase change in a complex, possibly haplo-diplontic, life-cycle. Seven of these associations are composed of heterococcoliths with holococcoliths; five of them are presented here for the first time: Helicosphaera carteri with Syracolithus catilliferus, Syracosphaera pulchra with Calyptrosphaera oblonga, Syracosphaera anthos with Periphyllophora mirabilis, Acanthoica quattrospina with holococcolithophorid sp. and Syracosphaera sp. with Corisphaera sp. type A (see Kleijne, A., 1991. Holococcolithophorids from the Indian Ocean, Red Sea, Mediterranean Sea and North Atlantic Ocean. Mar. Micropaleontol. 17, 1–76); the other two are Coronosphaera mediterranea with Calyptrolithina wettsteinii, found previously by Kamptner (Kamptner, E., 1941. Die Coccolithineen der Südwestküste von Istrien. Naturhistorischen Museum in Wien. Annalen 51, 54–149), and Syracosphaera nana with undescribed holococcoliths, figured previously by Kleijne (1991) as Syracosphaera sp. type A. In addition the association of Neosphaera coccolithomorpha and Ceratolithus cristatus, recently presented by Alcober and Jordan (Alcober, J., Jordan, R.W., 1997. An interesting association between Neosphaera coccolithomorpha and Ceratolithus cristatus (Haptophyta). Eur. J. Phycol. 32, 91–93) is verified by discovery of a further example. A further five combinations of heterococcoliths and holococcoliths are shown, these probably mostly represent life-cycle combinations but the evidence in each case is insufficient to consider these as definite associations although in these collapsed coccospheres the association of different species is less certain. Two examples of holococcolith–holococcolith associations are presented: S. catilliferus with Syracolithus confusus, and Zygosphaera bannockii with Corisphaera sp. type A. These are probably examples of intra-specific variation.A new species, Syracosphaera delicata sp. nov., is described.     

Seasonal variation in Emiliania huxleyi coccolith morphology and calcification in the Aegean Sea (Eastern Mediterranean)

A seasonal morphological variability is observed in Emiliania huxleyi var. huxleyi specimens, collected from discrete water samples in the Aegean Sea. Biometric analyses reveal a consistent pattern of increase in the size of coccoliths and coccospheres, including the thickness of the inner tube elements (INT), in winter/spring time low sea surface temperature and moderate productivity samples when compared with summer time high temperature-low productivity samples. The small range of salinity change in the Aegean Sea and the absence of seasonal pattern in nutrient content do not support any association with the observed increase in E. huxleyi coccolith size. A relatively increased [HCO₃^-] content is observed during spring-time interval related with the increase in the coccolith size, however it remains unclear which parameter of the carbonate system causes the observed effects.     

Light and electron microscope observations of the type species of Syracosphaera,s. pulchra (Prymnesiophyceae)

Syracosphaera pulchra, the type species of the genus was isolated into unialgal culture and studied with both the light and electron microscope. A conspicuous coiling haptonema is present containing seven microtubules in the shaft and eight in the basal region; features shared with many taxa in the order Prymnesiales. The proximal and distal coccoliths differ in shape but resemble each other structurally: the outer elements alternate to make the rim. The proximal coccoliths possess an organic base-plate scale which is absent in the distal coccoliths. The uncalcified organic scales are ornamented by a radial, more or less concentric, fibrillar pattern and are arranged in several layers between the proximal coccoliths and the plasmalemma. The ultrastructure of the cell is typical of prymnesiophycean algae. The flagellar apparatus is characterized by the absence of secondary microtubular bundles which are usually well developed in other coccolithophorids with two microtubular roots. This feature is also rather similar to that found in members of the Prymnesiales.

This investigation has indicated that S. pulchra has, in some respects, a closer affinity with members of the Prymnesiales than with the coccolithophorids.     

COCCOLITH FUNCTION AND MORPHOGENESIS: INSIGHTS FROM APPENDAGE‐BEARING COCCOLITHOPHORES OF THE FAMILY SYRACOSPHAERACEAE (HAPTOPHYTA)1

The morphology of three remarkable genera of coccolithophores, Ophiaster, Michaelsarsia, and Calciopappus, is reviewed based on new images using field emission scanning electron microscopy. Each of these genera characteristically forms coccospheres with long appendages formed of highly modified coccoliths, which radiate from either the circum‐flagellar pole of the coccosphere (Calciopappus and Michaelsarsia) or the antapical pole (Ophiaster). For each genus, it is shown that the appendage coccoliths can also occur in an alternative orientation appressed to the main coccosphere. It is hypothesized that the appendage coccoliths are initially deployed in the appressed orientation and that extension of the appendages is a dynamic response to environmental stress. The observations suggest that coccoliths are more sophisticatedly adapted to specific functions than has been assumed and that the cytoskeleton plays more active roles in coccolith morphogenesis and deployment than has previously been inferred.     

Disintegration of aggregates and coccospheres in sediment trap samples

A method for non-destructive disaggregation of coccoliths contained in fecal pellets, marine snow and on coccospheres is described. It consists of repeated chemical oxidation of organic material with (NaClO and H₂O₂) combined with brief periods of ultrasonification.     

Coccolithophore (–CaCO3) flux in the Sea of Okhotsk: seasonality, settling and alteration processes

Coccolithophore fluxes were determined in the Sea of Okhotsk using samples from a 1 year experiment (12 August 1990 to 12 August 1991) with sediment traps at 258 and 1061 m depth. A special study was made on Coccolithus pelagicus, using fragmentation and the degree of etching, as indicators of transport mechanisms. A Corrosion Index for C. pelagicus is developed. The coccolithophore flux pattern at 258 m depth was characterised by a strong seasonality, with flux peaks during autumn 1990 (late November to early December) and spring 1991 (March). The assemblage consisted almost entirely of the two species C. pelagicus and Emiliania huxleyi. During autumn, coccolithophore transportation to 258 m depth mainly occurred within cylindrical fecal pellets and marine snow aggregates of silicoflagellates, and through agglutination on tintinnids. Grazing caused severe fragmentation of coccoliths and disintegration of coccospheres. Marine snow aggregates contained many intact coccospheres of C. pelagicus. During spring, coccolithophores were probably removed from the euphotic zone by the ballast effect of sinking diatoms. The coccolithophore flux peak in spring occurred immediately after the ice had retreated from the trap station, and the trapped assemblage included coccoliths of subtropical species. These features indicate drifting from an ice-free location to the south or east.The coccolith and coccosphere flux at 1061 m was respectively 7 and 12 times lower than at 258 m depth, and maximum fluxes were recorded 2 months later. Increasing carbonate dissolution from 258 to 1061 m depth is expressed in the coccolithophore–CaCO₃ flux reduction of 82%, and in the increasing percentage of etched coccoliths of Coccolithus pelagicus from 32 to >90%.     

INDIAN OCEAN NANOPLANKTON. I. RHABDOSPHAERACEAE (PRYMNESIOPHYCEAE) WITH A REVIEW OF EXTANT TAXA1

Family characteristics of the Rhabdosphaeraceae are revised to limit the genera to those having cyrtoliths; genera with placoliths are removed from the family. Rhabdoliths, cyrtoliths bearing a process in the central area, are present in all genera. Coccospheres having monomorphic coccoliths, all being rhabdoliths, form one group within the family, whereas genera with dimorphic coccoliths in the coccosphere comprise a second group. Cyrtoliths without processes in the latter group may be intermixed with rhabdoliths in some genera, whereas other genera have rhabdoliths located only in polar regions of the coccosphere. Two generic nomenclatural changes are proposed, Algirosphaera being the name applied to species previously placed in Anthosphaera, an invalid generic name, and Palusphaera is recognized as a valid monotypic genus, P. vandeli being the name applied to the species that has been named Rhabdosphaera longistylis in recent literature. A new combination is made, Rhabdosphaera xiphos (Deflandre et Fert) comb. nov., recognizing a species formerly known only in sediments as extant. Rhabdosphaera, Acanthoica and Algirosphaera are genera with dimorphic coccoliths in the coccospheres; Discosphaera, Palusphaera and Anacanthoica are genera having monomorphic coccoliths in the coccosphere.     

Export production of coccolithophores in an upwelling region: Results from San Pedro Basin, Southern California Borderlands

A seven month-long time series sediment trap project was carried out in San Pedro Basin (Southern California Borderlands) in order to evaluate the response of calcareous nannoplankton to seasonal hydrographic changes. This region is periodically influenced by upwelling, particularly during the spring and early summer. The highest fluxes of both whole coccospheres and individual coccoliths occurred during winter (January-February), a period when the fluxes of diatoms and planktic foraminifera were low. The highest coccolithophore fluxes were recorded in the mid-February with 860 × 10⁶ coccoliths m⁻² day⁻¹, 8 × 10⁶ whole coccospheres m⁻² day⁻¹, and 80 mg of coccolith carbonate m⁻² day⁻¹. Coccolith carbonate fluxes in January and February account for most of the total carbonate fluxes measured during this period. The season of maximum coccolithophore production in this region (winter) is correlated with weak stratification of the upper water column, low total primary production, low nutrient contents, and low temperatures.Emiliania huxleyi and Florisphaera profunda are the two most abundant species in this region. While E. huxleyi displays no distinct seasonal changes in flux, F. profunda shows a clear preference for cold, low nutrient water conditions and low light levels. Helicosphaera spp. flux is positively correlated to the total coccosphere fluxes and is indicative of high coccolithophore productivity.     

Seasonal occurrence of coccoliths in sediment traps from West Caroline Basin, equatorial West Pacific Ocean

Coccolith fluxes were investigated by sediment trap studies in the West Caroline Basin, which is located in the equatorial western Pacific. The investigation was conducted from June 1991 to March 1992 at two water depths, 1592 and 3902 m, as part of the Northwest Pacific Carbon Cycle Study (NOPACCS) program. Two seasonal maxima of coccolith fluxes were observed during September–early October and late December–January. The average coccolith and coccosphere fluxes at the depth of the shallow trap were 1800×10⁶ coccoliths m⁻² day⁻¹ and 1.9×10⁶ coccospheres m⁻² day⁻¹, respectively. The flux of coccoliths followed the same trend as the total flux, and was closely correlated with the flux of organic matter flux. Florisphaera profunda, Gladiolithus flabellatus, Gephyrocapsa oceanica, Umbilicosphaera sibogae var. sibogae, Emiliania huxleyi, and Oolithotus fragilis were the most abundant species together comprising more than 85% of the total flora. Observed seasonal changes of the species composition of coccolith flora, as well as analysis of the R-mode cluster, revealed that during the summer, the assemblage was marked by the dominance of G. oceanica and U. sibogae. However, during the winter, the assemblage was dominated by E. huxleyi and O. fragilis. These assemblage changes were influenced by monsoonal events, which were observed off the New Guinea coast. F. profunda dominated the community in the shallow trap throughout most of the year; peak values of this species were recorded during the winter. The coccosphere assemblage was dominated by G. oceanica at both water depths. In the deep trap, the sedimentation pattern was similar to that observed at the shallow depth. Mean coccolith and coccosphere fluxes at the deep trap were 2000×10⁶ coccolith m⁻² day⁻¹ and 0.08×10⁶ coccospheres m⁻² day⁻¹, respectively. The increase in coccolith flux with water depth suggests a lateral influx. The estimated average daily mass of CaCO₃ flux in coccoliths and coccospheres was 16.6 mg m⁻² day⁻¹ at the 1592 m trap and 17.9 mg m⁻² day⁻¹ at the 3902 m trap, respectively. These calculated values contributed only 23.3% to the total CaCO₃ flux at the shallow trap and 27.9% at the deep trap.     

LA ENPRENO DE STRONTIO EN KOKOLITOFOROJ

Pure cultures of the coccolithophorid Syracosphaera carleraewere grown in a synthetic saline medium containing 3.4mM Ca++and 2.0, 1.0, 0.5 or 0.0 mM Sr⁺⁺. The coccoliths were separatedby differential centrifugation, washed, dried, and examinedby flame photometry and by X-ray diffraction. In the absenceof Sr, they consisted of pure calcite. In media containing Sr,the concentration factor or incorporation factor (Sr/Ca in coccolithsSr/Ca in medium) was approx. 0.02 in each case, indicating ahigh degree of discrimination against Sr. ¹ Nuna adreso: Scripps Instituto por Oceanografio, La Jolla,California, U. S. A. (Received March 4, 1961; )     

Life-cycle associations involving pairs of holococcolithophorid species: intraspecific variation or cryptic speciation?

New holococcolith-heterococcolith life-cycle associations are documented based on observations of combination coccospheres. Daktylethra pirus is shown to be a life-cycle phase of Syracosphaera pulchra and Syracolithus quadriperforatus a life-cycle phase of Calcidiscus leptoporus. In addition, new observations from cultures confirm the life-cycle associations of Crystallolithus braarudii with Coccolithus pelagicus and of Zygosphaera hellenica with Coronosphaera mediterranea. In all four cases previous work has shown that the heterococcolithophorid species is associated with another holococcolithophorid. Two other examples of a heterococcolithophorid being associated with two holococcolithophorids have previously been identified, so this seems to be a common phenomenon. The six examples are reviewed to determine whether a single underlying mechanism is likely to be responsible for all cases. It is concluded that there is no single mechanism but rather that the six examples fall into three categories: (a) in two cases the holococcolith types are probably simply ecophenotypic morphotypes; (b) in two other cases the holococcolith types are discrete and are paralleled by morphometric differences in the heterococcolith types; (c) in the final two cases the holococcolith types are discrete but are not paralleled by any obvious morphological variation in the heterococcolith morphology. We infer that cryptic speciation may be widespread in heterococcolithophorid phases and that study of holococcolithophorid phases can provide key data to elucidate this phenomenon.     

Mg isotopes and Mg/Ca values of coccoliths from cultured specimens of the species Emiliania huxleyi and Gephyrocapsa oceanica

Coccoliths from cultured specimens of two species of coccolithophores (Emiliania huxleyi and Gephyrocapsa oceanica) were sampled during two growth phases (late exponential and stationary), and their Mg isotope values (δ²⁶Mg) as well as Mg/Ca values were measured in order to investigate whether δ²⁶Mg can be used as a temperature proxy. Mg/Ca values were positively related with temperature (~ 0.002 mmol/mol/°C), without statistically significant differences between the two growth phases and the two species. Both species were depleted in heavier Mg isotopes relative to the culture medium, and δ²⁶Mg values were temperature dependent in both growth phases of E. huxleyi, although the δ²⁶Mg values differed in the two growth phases. In G. oceanica, a weak correlation between δ²⁶Mg values and temperature was seen in the late exponential growth phase only, and the δ²⁶Mg values differed between growth phases. The large differences between δ²⁶Mg values as measured in calcite formed during different growth phases indicate that Mg isotopes of coccoliths cannot be simply used as a temperature proxy. Our conclusions are preliminary and more data must be collected in order to fully evaluate the use of Mg isotopes of coccoliths as a temperature proxy.     

Incorporation of zinc into the coccoliths of the microalga <Emphasis Type="Italic">Emiliania huxleyi</Emphasis>

The coccolithophore Emiliania huxleyi is covered with elaborated calcite plates, the so-called coccoliths, which are produced inside the cells. We investigated the incorporation of zinc into the coccoliths of E. huxleyi by applying different zinc and calcium amounts via the culture media and subsequently analyzing the zinc content in the cells and the Zn/Ca ratio of the coccoliths. To investigate the Zn/Ca ratio of coccoliths built in the manipulated media, the algae have first to be decalcified, i.e. coccolith free. We used a newly developed decalcification method to obtain ‘naked’ cells for cultivation. E. huxleyi proliferated and produced new coccoliths in all media with manipulated Zn/Ca ratios. The cells and the newly built coccoliths were investigated regarding their zinc content and their Zn/Ca ratio, respectively. High zinc amounts were taken up by the algae. The Zn/Ca ratio of the coccoliths was positively correlated to the Zn/Ca ratio of the applied media. The unique feature of the coccoliths was maintained also at high Zn/Ca ratios. We suggest the following pathway of the zinc ions into the coccoliths: first, the zinc ions are bound to the cell surface, followed by their transportation into the cytoplasm. Obviously, the zinc ions are removed afterwards into the coccolith vesicle, where the zinc is incorporated into the calcite coccoliths which are then extruded. The incorporation of toxic zinc ions into the coccoliths possibly due to a new function of the coccoliths as detoxification sites is discussed.     

Patterns of carbon and nitrogen uptake during blooms of Emiliania huxleyi in two Norwegian fjords

Blooms of the coccolithophorid Emiliania huxleyi were monitoredin two land-locked fjords, Fauskangerpollen and Nordåsvannet(Western Norway), in May 1993. The chemical composition of particulatematter, size-fractionated photosynthesis, calcification, nitrogenuptake rates and the patterns of macromolecular synthesis wereexamined during the peak and decline of E.huxleyi blooms. Thetemporal evolution of the bloom in Fauskangerpollen was definedby a gradual decrease in cell abundance and cell-specific calcificationrates, and by increasing numbers of empty coccospheres and theratio detached coccoliths/living cells. A large proportion ofthe nitrogen required for microplankton growth was suppliedby aminonium and dissolved organic compounds such as urea and,as a consequence, the f-ratios were low (0.2). In general, nitrogenuptake patterns were consistent with ambient concentrationsof nitrogenous species. The photosynthetic carbon metabolismof E.huxleyi dominated phytoplankton assemblages was characterizedby high carbon allocation into lipids and relatively low carbonincorporation into protein as compared with diatom-dominatedassemblages. Consequently, the organic C/nitrogen uptake ratiodetermined stoichiometrically was significantly higher (up to10.8) when coccolithophorids were dominant than in diatom-basedor mixed-phyto-plankton assemblages. These carbon incorporationpatterns were reflected in differences in the chemical compositionof particulate matter.     

2010秋季东海今生颗石藻的空间分布

根据2010年11月东海29站位所采集108个样品偏光镜检分析结果显示:调查区共发现26种今生颗石藻,优势种为赫氏艾密里藻(Emiliania huxleyi)、大洋桥石藻(Gephyrocapsa oceanica)、卡特螺旋球藻(Helicosphaera carteri)和粗壮环翼球(Algirosphaera robusta)等.水体中今生颗石藻丰度为0-76.562个/mL,平均值为18.641个/mL;颗石粒丰度0-4506.47个/mL,平均值为613.44个/mL,今生颗石藻在表层和底层均呈斑块状分布,海盆区站位丰度较近岸站位下降明显,但物种丰富度明显增多;颗石球丰度最大值站位发生在硅藻水华站位.将研究区域划分为3个断面进行分析:PN断面、沿岸流断面和黑潮断面.比较显示,PN断面的空间分布差异性较大,赫氏艾密里藻具有最高丰度的颗石粒和颗石球.沿岸流断面颗石粒/颗石球、黑潮断面中颗石粒(除底层外),自表至底随水深增加有轻微增加趋势,黑潮断面颗石球丰度高值区集中于25-80 m之间.结合历史环境资料,分析得出东海调查期水体混合、泥沙再悬浮以及硝酸盐浓度是控制今生颗石藻物种丰富度与物种特异分布的主要因子;今生颗石藻在不同环境中具有不同的生活策略,其空间分布特征随取样季节和海区不同有较大变化.     

Further observations on Vexillarius cancellifer Jordan & Chamberlain

A single specimen of an unusual dimorphic coccosphere was encountered in the subtropical North Atlantic. Despite its poor condition, it was formally described in 1993 as a new lower photic zone species, Vexillarius cancellifer Jordan & Chamberlain. Since then, the species has only been reported twice, with little or no additional information to the original diagnosis. In 2005, a new specimen was found in the Java upwelling system in the southeastern Indian Ocean, and like the type specimen, it was collected from the lower photic zone. The distal portions of the tubular coccoliths are far more complete in the new specimen. We therefore provide an emended diagnosis for this rare genus and species.     

Coccolith Sr/Ca ratios in the eastern Mediterranean: Production versus export processes

Samples collected by two sediment traps located southwest of Crete in the eastern Mediterranean (EMED) [48A (1953 m) and 48B (950 m)] from June 2005 to May 2006 were used to study fluxes of organic carbon, carbonate and coccolithophores in combination with the variations of Sr/Ca ratios in different individually picked coccolith species. Considering the complexity of the EMED, we validate the use of Sr/Ca ratios as productivity proxy and unravel the varied processes which may influence it. We examined the relationship between the seasonal peaks in export fluxes and the Sr/Ca ratio in coccoliths of three upper photic zone coccolithophores species collected in the traps, Calcidiscus leptoporus, Helicosphaera carteri and Emiliania huxleyi. We aimed at testing whether high export fluxes are correlated with high Sr/Ca ratios, suggestive of higher nutrient-stimulated production, or Sr/Ca ratios are unchanged during high export periods, suggestive of increased export efficiency or scavenging. Periods of enhanced trap fluxes in March and June result from surface water blooms recognized in satellite imagery. An additional peak flux was found in January, but this peak represents re-suspended or recycled material in the water column.The amplitude of seasonal variations in the Sr/Ca ratios of the three investigated species is small in both traps. In the shallow trap, a decrease in the Sr/Ca ratio of C. leptoporus occurred synchronously with minimal fluxes. The other two species were not measured for this period. In the deep trap, no such decrease in Sr/Ca was observed during minimal fluxes, in either C. leptoporus or H. carteri, probably due to a long residence of coccoliths in the water column, recycling and low export efficiency. Absolute Sr/Ca ratios for all species are lower than in other more productive environments like the Bay of Bengal, Arabian Sea, or Sargasso Sea. We conclude that Sr/Ca ratios in coccoliths of surface sediments in the EMED reflect mainly spring–summer bloom conditions averaged over hundreds to thousands of years.In addition, the origin of varying calcite thickness in H. carteri was investigated. The similarity of average Sr/Ca ratios in differently-calcified specimens confirms that coccolith thickness variations in this species result from primary biomineralization processes and not from variable overgrowth by (low Sr) abiogenic calcite in the water column or the sediments.     

COCCOLITH PRODUCTION AND DETACHMENT BY EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE)1

Laboratory experiments were performed with the prymnesiophyte Emiliania huxleyi (Lohm.) Hay and Mohler, strain 88E, to quantify calcification per cell, coccolith detachment, and effects of coccolith production on optical scattering of individual cells. ¹⁴C incorporation into attached and detached coccoliths was measured using a bulk filtration technique. ¹⁴C-labeled cells also were sorted using a flow cytometer and analyzed for carbon incorporation into attached coccoliths. The difference between the bulk and flow cytometer analyses provided a ¹⁴C-based estimate of the rate of production of detached coccoliths. Coccolith production and detachment were separated in time in batch cultures, with most detachment happening well after calcification had stopped. Accumulation of coccoliths was maximum at the end of logarithmic growth with 50–80 coccoliths per cell (three to five complete layers of coccoliths around the cells). Net accretion rates of coccoliths were on the order of 7 coccoliths· cell^?1·d^?1 while net detachment rates were as high as 15 coccoliths· cell^?1·d^?1 for stationary phase cells. Equal numbers of coccoliths were attached and detached early in logarithmic growth, and as cells aged, the numbers of detached coccoliths exceeded the attached ones by a factor of 6. Our results demonstrate pronounced charges of forward angle light scatter and 90° light scatter of cells as they grow logarithmically and enter stationary phase. Counts of loose coccoliths in batch cultures are consistent with the detachment of coccoliths in layers rather than individual coccoliths.     

Effects of cytochalasin B on division and calcium carbonate extrusion in a calcareous alga.

Cytochalasin B (CB) (100 μg/ml) reversibly blocked cell division and cuased the formation of abnormal cytoplasmic bodies in the alga Cricosphaera carterae. Concentrations of 20 μg/ml and 40 μg/ml CB were without effects. In the presence of CB, calcified bodies (coccoliths) which form in Golgi vesicles and are normally extruded through the plasma membrane were not extruded and accumulated within the cell. CB appeared to alter the membranes of Golgi vesicles containing coccoliths. DMSO (10% $\text{v}\text{v}$), the solvent for CB, was without effect on cell division and coccolith extrusion. A concentration of 20% $\text{v}\text{v}$ DMSO inhibited cell division irreversibly.