Seasonal variations in the morphology of the coccolithophore Calcidiscus leptoporus off Bermuda (N. Atlantic)

Calcidiscus leptoporus is a cosmopolitan coccolithophore specieswith one of the longest fossil records. A previous morphometricstudy on coccoliths of C. leptoporus, carried out on a globalset of Holocene sediment samples, indicated that temperaturemight be a significant factor influencing the size distributionof C. leptoporus. In the present study, C. leptoporus morphologicalvariations and dynamics are studied in samples from HydrostationS time series, near Bermuda, over a 3 year interval (1991 to1994) with monthly sampling. Calcidiscus leptoporus was presentall year round in very low abundances but displayed strong variationsthrough the time-series, both in absolute abundances (maximum551 cells/l and minimum <10 cells/l) and in morphology. Nosignificant correlation between environmental parameters andC. leptoporus abundances was found. The seasonal dynamics ofC. leptoporus, therefore, appears to be the consequence of planktoncommunity interactions and, possibly, of life-cyle. The morphologicalstudy of C. leptoporus shows a sharp decrease in size and variabilityoccurring during the spring–summer transition at the onsetof water stratification. Our results give no conclusive evidenceas to whether C. leptoporus at Hydrostation S consists of anassemblage of different species or ecophenotypes. However, weprovide evidence for significant morphological plasticity linkedto seasonality and possibly temperature. More importantly, ourresults show a trend between temperature and morphology oppositeto the study of Knappertsbusch et al. (Knappertsbusch et al.,1997) based on Holocene sediments. These results emphasize thenecessity of plankton studies in order to interpret patternsobserved in sediments.     

Morphologic variability of the coccolithophorid Calcidiscus leptoporus in the plankton, surface sediments and from the Early Pleistocene

On a global scale, morphological variability of the extant coccolithophorid Calcidiscus leptoporus (Murray and Blackman, 1898) Loeblich and Tappan was investigated in surface sediments and plankton samples and from an Early Pleistocene time-slice (1.8 Ma to 1.6 Ma). In the bivariate space coccolith diameter versus number of rays in the distal shield, Holocene samples follow a single, unimodal morphocline. Sample means of coccolith size and number of elements group in three clusters, I, II and III, which are of biogeographic significance. Clusters II and III coccoliths (mean coccolith size of 5.0 μm and 20.9 elements, and 6.6 μm and 25.6 elements, respectively) are found in a tropical belt extending from 11 °N to 17 °S with an annual minimum sea-surface temperature above 23.5 °C. Cluster I coccoliths (5.8 μm, 20.7 elements) are found in samples outside that belt. The distribution of coccoliths in the surface sediments is tentatively interpreted to be a result of mixing to a varying degree of at least three different morphotypes (‘small’, ‘intermediate’ and ‘large’), which were identified in the living plankton, and which are separated from each other at 5 μm and 8 μm mean coccolith diameter, respectively. A comparison of the surface sediments with the Early Pleistocene assemblages revealed that between 1.6 Ma and 1.8 Ma two morphoclines A and B existed, the first of which persisted until the Holocene in the form of C. leptoporus, while the second comprises only extinct morphotypes including Calcidiscus macintyrei as one end-member. During the Early Pleistocene morphocline A was more homogeneous and no clusters were evident.Morphocline B shows a clear bimodality with a separation of morphotypes at 9.5 μm. Our observations suggest that morphoclines are subsets within the total stratigraphical range of a single species, and represent the global variability of that species in a particular time interval. Morphotypes, which belong to a morphocline, represent the infra-specific variability of that species within the biogeographic and stratigraphic limits of that species.     

Observations on Syracosphaera rhombica sp. nov.

A spherical coccosphere and two collapsed coccospheres composed of monomorphic rhombic coccoliths were encountered in 2005 in the Java upwelling system of the SE Indian Ocean, while a further two specimens with elongate coccospheres were recently found in the Gulf of Mexico. All of the specimens were collected from the lower photic zone (75–160 m). The coccoliths possess a proximal flange, a slightly flared wall with a serrated distal margin, and a relatively plain central area structure comprised only of overlapping laths. The taxon appears to be an undescribed species of the Syracosphaera nodosa group, so we describe it herein as Syracosphaera rhombica sp. nov.     

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.     

Holococcolithophore‐heterococcolithophore (Haptophyta) life cycles: Flow cytometric analysis of relative ploidy levels

Several coccolithophore species are known to exhibit heteromorphic life cycles. In certain species, notably Emiliania huxleyi, the heterococcolith‐bearing phase alternates with a non‐calcifying stage, whereas in others the heterococcolith‐bearing phase alternates with a holococcolith‐bearing phase. Heterococcolithophore‐holococcolithophore life cycles have previously been observed for only one species in culture, but have also been inferred from an increasing number of observations of combination coccospheres. 18S rDNA sequences from pure cultures of both the heterococcolith‐bearing and holococcolith‐bearing phases of Coccolithus pelagicus were identical, providing an additional indication of their identity as different life cycle stages of the same species. Flow cytometric analyses have been undertaken on SybrGreen‐stained nuclei isolated from pure cultures of the two phases of four coccolithophore species (Coccolithus pelagicus, Calcidiscus leptoporus, Coronosphaera mediterranea and Emiliania huxleyi) in order to determine relative DNA content. Results confirm the hypothesis that holococcolithophore‐heterococcolithophore life cycles are haplo‐diploid in nature. Light microscope observations of the processes of sexual fusion and meiosis are reported for two of the experimental species. The results are discussed in the context of the evolution of bio‐mineralization in the coccolithophores and the possible ubiquity of haplo‐diploidy in the haptophytes.     

Crepidolithus cantabriensis nov. sp., a new calcareous nannofossil (Prymnesiophyceae) from the Lower Jurassic of northern Spain

A new calcareous nannofossil, Crepidolithus cantabriensis nov. sp., is described from the Lower Jurassic marls and marly limestones of the Cantabrian Range (Northern Spain). The new species is characterized by medium-sized, normal to narrowly elliptical coccoliths with open central-areas. In the light microscope, under crossed nicols and in distal view, it displays a characteristic bicyclic extinction pattern. Its inner rim edge shows maximum birefringence colours, which vary from white to yellow, whereas the faintly outer rim margin exhibits low birefringence, tending to grey, and they are separated by a dark and sigmoidal isogyre. C. cantabriensis nov. sp. seems closely related to Crepidolithus crassus on the basis of shared morphological characters. Both of them have a distal shield composed of around thirty sub-vertical, dextrally imbricating laths. The central-area width of C. cantabriensis nov. sp. is approximately half of the coccolith width, whilst it is reduced to one or two tiny lenticular slits in C. crassus. Nevertheless, the phyletic relationships between C. cantabriensis nov. sp. and Crepidolithus cavus are not clear, since they show different rim structures, and C. cavus has a delicate bar aligned along the minor axis of its central area. The first specimens of C. cantabriensis nov. sp. have been recognized within the Raricostatum AZ in the Tudanca section. This species seems to be more abundant in the Reinosa area, which occupied a more distal position with respect to the Iberian Massif during the studied time interval. It is hypothesized here that C. cantabriensis nov. sp. could have inhabited the lower-photic zone.     

Movement of oceanic fronts south of Australia during the last 10 ka: interpretation of calcareous nannoplankton in surface sediments from the Southern Ocean

Preservation of calcareous nannoplankton in surface sediment samples from the Southern Ocean south of Australia and adjacent to New Zealand record a single assemblage. The dominant species are Emiliania huxleyi, Gephyrocapsa muellerae, Calcidiscus leptoporus, Helicosphaera carteri and Coccolithus pelagicus. The assemblage varies little in abundance and diversity with minor correlation to present-day overlying surface water masses and oceanic fronts. Increase in abundance of H. carteri and C. pelagicus in the region of the Subtropical Front may reflect higher nutrients associated with this front. The assemblage, although altered by dissolution, represents a warmer climatic interval than present-day with the presence of preferentially dissolved, warm-water species preserved as far south as the Polar Front. The presence of warm-water species under sub-Antarctic waters at the Polar Front is interpreted as a relic population from the Holocene climatic optimum of 10–8 ka. The absence of coccoliths in sediments poleward of the Polar Front suggests an equatorward shift of this front following the climatic optimum, resulting in increased productivity of siliceous phytoplankton associated with the colder waters and increased dissolution of coccoliths. Movement of the Subtropical Front for the same interval is not recorded in the preserved coccoliths. The more heavily calcified form of E. huxleyi which dominates the living assemblage north of the Subtropical Front is subject to dissolution in this region and is poorly preserved in the sediment assemblage.     

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.     

Molecular evolution of the CMT1A-REP region: a human- and chimpanzee-specific repeat.

The CMT1A-REP repeat consists of two copies of a 24-kb sequence on human chromosome 17p11.2-12 that flank a 1.5-Mb region containing a dosage-sensitive gene, peripheral nerve protein-22 (PMP22). Unequal meiotic crossover mediated by misalignment of proximal and distal copies of the CMT1A-REP in humans leads to a 1.5-Mb duplication or deletion associated with two common peripheral nerve diseases, Charcot-Marie-Tooth disease type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP). Previous molecular hybridization studies with CMT1A-REP sequences suggested that two copies of the repeat are also found in the chimpanzee, raising the possibility that this unique repeat arose during primate evolution. To further characterize the structure and evolutionary synthesis of the CMT1A-REP repeat, fluorescent in situ hybridization (FISH) analysis and heterologous PCR-based assays were carried out for a series of primates. Genomic DNA was analyzed with primers selected to differentially amplify the centromeric and telomeric ends of the human proximal and distal CMT1A-REP elements and an associated mariner (MLE) sequence. All primate species examined (common chimpanzee, pygmy chimpanzee, gorilla, orangutan, gibbon, baboon, rhesus monkey, green monkey, owl monkey, and galago) tested positive for a copy of the distal element. In addition to humans, only the chimpanzee was found to have a copy of the proximal CMT1A-REP element. All but one primate species (galago) tested positive for the MLE located within the CMT1A-REP sequence. These observations confirm the hypothesis that the distal CMT1A-REP element is the ancestral sequence which was duplicated during primate evolution, provide support for a human-chimpanzee clade, and suggest that insertion of the MLE into the CMT1A-REP sequence occurred in the ancestor of anthropoid primates.     

HYMENOMONAS CORONATA SP. NOV., A NEW COCCOLITHOPHORID FROM THE TEXAS COAST1,2

A new species of Hymenomonas (H. coronata) has been isolated from the Texas coast, grown in axenic culture, and examined by both light and electron microscopy. Certain cultural and ultrastructural observations are presented. This alga has several haptophycean characteristics, although a definite haptonema has not been observed. Similarities to other members of the genus are evident, such us type and shape of coccoliths and pyrenoid-chloroplast morphology.     

A complex role for distal-less in crustacean appendage development.

The developing leg of Drosophila is initially patterned by subdivision of the leg into proximal and distal domains by the activity of the homeodomain proteins Extradenticle (Exd) and Distal-less (Dll). These early domains of gene expression are postulated to reflect a scenario of limb evolution in which an undifferentiated appendage outgrowth was subdivided into two functional parts, the coxapodite and telopodite. The legs of most arthropods have a more complex morphology than the simple rod-shaped leg of Drosophila. We document the expression of Dll and Exd in two crustacean species with complex branched limbs. We show that in these highly modified limbs there is a Dll domain exclusive of Exd but there is also extensive overlap in Exd and Dll expression. While arthropod limbs all appear to have distinct proximal and distal domains, those domains do not define homologous structures throughout arthropods. In addition, we find a striking correlation throughout the proximal/distal extent of the leg between setal-forming cells and Dll expression. We postulate that this may reflect a pleisiomorphic function of Dll in development of the peripheral nervous system. In addition, our results confirm previous observations that branch formation in multiramous arthropod limbs is not regulated by a simple iteration of the proximal/distal patterning module employed in Drosophila limb development.     

Two new species of Mallomonas (Synurophyceae) from the Ocala National Forest, Florida, U.S.A.

Two new species of Mallomonas, M. ocalensis and M. caerula , are described from a clearwater, acidic and oligotrophic waterbody in the Ocala National Forest, Florida, U.S.A. Mallomonas ocalensis , in the Series Doignonianae of the Section Torquatae, consists of relatively small cells with scales that have a series of parallel shield ribs with alternating thicknesses, collar scales with very short bristles and posterior scales with small protruding spines. Mallomonas ocalensis is most similar to Mallomonas dickii , but is distinguished from the latter species based on the highly variable thicknesses of the shield ribs and significantly larger scales. Mallomonas caerula , in the Series Mallomonas of the Section Mallomonas, consists of relatively large cells that bear large domed and dorneless scales, and long, robust and ribbed bristles. Bristles with and without helmets can be found. The shield, especially the distal portion, and the posterior flange of scales consists of series of more or less parallel ribs. Mallomonas caerula appears to be most closely related to Mallomonas acaroides var. acaroides and Mallomonas acaroides var. muskokana , but can be distinguished from the latter taxa on the basis of scale type, scale structure and bristle morphology. Both new species were found in the plankton and surface sediments of Blue Sink Pond.     

Coccolith biomineralisation studied with atomic force microscopy

Biomineralisation can only be understood as an interplay between organic and mineral phases. With this objective, we conducted an investigation of coccoliths using atomic force microscopy (AFM), an ultra-high resolution technique that requires no surface coating and can be used in air or under solution at ambient conditions of temperature and pressure. The detailed morphology, crystal structure, organic scales and organic coating of the coccolith species Coccolithus pelagicus , Helicosphaera carteri and Oolithotus fragilis were investigated. The fine structure of coccoliths is very complex, with the calcite either being smooth, dominated by steps or tuberculate; organic cover can be either granular or fibrous. Behaviour of coccolith surfaces during dissolution is influenced both by mineral and organic material and different surface types show variable resistance to dissolution. The organic coating protects element faces against etching. Through atomic resolution AFM, it is possible to establish the crystallographic structure of the distal shields of C. pelagicus and O. fragilis . Though elements of both species are dominated by stable crystal faces, there are important differences between them, with the external edge of elements being parallel to a cleavage direction in C. pelagicus but parallel to the atomic rows in O. fragilis . Thus, there is evidence that the biomineralisation of each species, and also of select areas of coccoliths of the same species, is markedly different.     

NEW EVIDENCE FOR MORPHOLOGICAL AND GENETIC VARIATION IN THE COSMOPOLITAN COCCOLITHOPHORE EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) FROM THE COX1b‐ATP4 GENES1

Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler is a cosmopolitan coccolithophore occurring from tropical to subpolar waters and exhibiting variations in morphology of coccoliths possibly related to environmental conditions. We examined morphological characters of coccoliths and partial mitochondrial sequences of the cytochrome oxidase 1b (cox1b) through adenosine triphosphate synthase 4 (atp4) genes of 39 clonal E. huxleyi strains from the Atlantic and Pacific Oceans, Mediterranean Sea, and their adjacent seas. Based on the morphological study of culture strains by SEM, Type O, a new morphotype characterized by coccoliths with an open central area, was separated from existing morphotypes A, B, B/C, C, R, and var. corona, characterized by coccoliths with central area elements. Molecular phylogenetic studies revealed that E. huxleyi consists of at least two mitochondrial sequence groups with different temperature preferences/tolerances: a cool‐water group occurring in subarctic North Atlantic and Pacific and a warm‐water group occurring in the subtropical Atlantic and Pacific and in the Mediterranean Sea.     

19F NMR of trifluoroacetyl-labeled cysteine mutants of myoglobin: structural probes of nitric oxide bound to the H93G cavity mutant.

Nitric oxide (NO) binds to the myoglobin (Mb) cavity mutant, H93G, forming either a 5- or 6-coordinate Fe--NO heme complex. The H93G mutation replaces the proximal histidine of Mb with glycine, allowing exogenous ligands to occupy the proximal binding site. In the absence of the covalently attached proximal ligand, NO could bind to H93G from the proximal side of the heme rather than the typical diatomic binding pocket on the distal side when the 5-coordinate complex forms. The question of whether NO binds on the distal or proximal side was addressed by (19)F NMR. Site-directed mutagenesis was used to introduce unique cysteine residues at the protein surface on either the distal (S58C) or proximal (L149C) side, approximately equidistant from and perpendicular to the heme plane of both wild-type and H93G Mb. The cysteine thiols were alkylated with 3-bromo-1,1,1-trifluoroacetone to attach a trifluoroacetyl group at the mutation site. (19)F NMR spectra of 5-coordinate, NO bound S58C/H93G and L149C/H93G double mutants depict peaks with line widths of 100 and 23 Hz, respectively. As fluorine peaks broaden with increasing proximity to paramagnetic centers, such as 5-coordinate Fe--NO, the (19)F NMR data are consistent with NO binding in the distal heme pocket of H93G, even in the absence of a sixth axial ligand. Additionally, (19)F NMR spectra are reported for deoxy, oxy, CO, met CN, and met H(2)O forms of the labeled cysteine mutants. These results demonstrate that the fluorine probes are sensitive to subtle conformational changes in the protein structure due to ligation and oxidation state changes of the heme iron in Mb.     

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.     

Innervation of the limb accessory flexor muscle in several decapod crustaceans. II. Electrophysiology

The innervation of the distal and proximal heads of the accessory flexor muscle in three portunid crabs and two non-portunid decapods was studied electrophysiologically. In all species studied, the proximal head received only the two previously reported accessory flexor axons, an excitor and an inhibitor. The same two axons also innervated the distal head in all species, but in the portunids the distal head also received excitation from at least three, and probably sometimes four, of the main flexor excitor efferents. The accessory inhibitor exerted very strong effects in the tonic muscle fibers found in the proximal head and in the most proximal bundle of the distal head. The newly described inhibitory and excitatory distributions may have important implications for locomotory behavior.     

Strontium/Calcium ratio, carbon and oxygen stable isotopes in coccolith carbonate from different grain-size fractions in South Atlantic surface sediments

In this study, the Strontium/Calcium (Sr/Ca) ratio, and the carbon and oxygen isotopic compositions of coccoliths are investigated in three different grain-size fractions (<20 μm, 15-5 μm, <5 μm) of 17 surface sediment samples from the Equatorial and South Atlantic. The results are compared to environmental parameters in order to assess the factors controlling the observed coccolith geochemical patterns. Isotopic and geochemical composition of coccolith species in surface sediment samples from the South Atlantic greatly varies according to the different grain-size fractions. However, even if the absolute values show a great offset, the general trends are comparable. The δ¹⁸O values show a decreasing trend with increasing temperature. The δ¹³C and Sr/Ca ratio are mainly influenced by productivity of coccolithophores, which is in turn controlled by different factors, such as temperature, nutrient supply and productivity of other phytoplankton groups. Dilution and dissolution are negligible factors in these open marine samples. Therefore, coccolith abundance in bulk sediment is the best approximation for productivity of coccolithophores. The various coccolith species fractionate Sr differently, as is best shown by the 5-15 μm fraction where three species (Calcidiscus leptoporus, Helicosphaera carteri and Coccolithus pelagicus) predominantly occur.