MOLECULAR PHYLOGENETIC ANALYSIS OF rbcL IN THE PRYMNESIOPHYTA1

The nucleotide sequences of rbcL genes encoding the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were determined from six species of Prymnesiophyta to clarify their phylogenetic relationships. Molecular phylogenetic trees were constructed using PAUP (Phylogenetic Analysis Using Parsimony). These analyses suggest that the Prymnesiophyta, except for the Pavlovales, area relatively stable monophyletic group. Pleurochrysis carterae, included in the Isochrysidales, is a sister species of a monophyletic group consisting of other members of the Isochrysidales, Gephyrocapsa oceanica and Emiliania huxleyi, members of the Coccosphaerales, Calyptrosphaera sphaeroidea and Umbilicosphaera sibogae var. foliosa, and a member of the Prymnesiales, Chrysochromulina hirta. The nucleotide sequence of rbcL from G. oceanica was identical to that from E. huxleyi within the region examined. Our trees show that G. oceanica and E. huxleyi are more closely related to C. hirta than to U. sibogae, C. sphaeroidea, and P. carterae. These results suggest that orders in the Prymnesiophyceae, including the above-mentioned genera, should be redefined.     

CARBONIC ANHYDRASE AND CARBON FIXATION IN COCCOLITHOPHORIDS1

Rates of carbon fixation in coccolithophorids in culture, unlike many other algae, are carbon limited at ambient levels of dissolved inorganic carbon (DIC). Apparently, plants often rely on activity of carbonic anhydrase (CA) to raise the level of CO₂ in cells and achieve carbon saturation. However, CA activities in the coccolithophorids, Coccolithus (= Emiliania) huxleyi Lohmann and Hymenomonas (=Cricosphaera) carterae Braarud, were either not detectable or very low compared to activities in other systems, including other algae, higher plants, and representative animals. Furthermore, additions of CA to medium with 2 mM DIC at pH 8.1 resulted in nearly 30% enhancement of photosynthesis, but not coccolith formation. Although carbon fixation in coccolithophorids can be suppressed by the CA inhibitor acetazolamide, studies of CaCO₃ nucleation revealed a non-specific effect of the inhibitor. Using a 30 min assay based on pH decreases accompanying loss of dissolved. CO₃^2-, inhibition of crystal formation in the absence of CA at 1 mM acetazolamide was demonstrated for decalcified crab carapace, a tissue with which normal CaCo₃ deposition in vitro has been shown. The results suggest only a minor role for CA in coccolithophorids.     

CALCIFICATION BY COCCOLITHOPHORIDS: EFFECTS OF pH AND Sr1

Cells of Coccolithus huxleyi which fail to deposit CaCO₃ and form coccoliths often occur as unwanted components in cultures used for studies of calcification. Non-calcified cells generally cannot be made to recalcify, but they can be removed from cultures by treatment at elevated pH or by a method based on faster sinking of calcified cells. Lowering the concentrations of nitrate, phosphate, or trace metals in the medium did not restore calcifying ability of non-calcified cells. However, addition of strontium did promote recalcification of decalcified Cricosphaera carterae grown under calcium limitation. Strontium seemed to promote coccolith attachment to cells rather than to affect calcium uptake or coccolith formation itself.     

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.     

LOCALIZATION OF CA2+-STIMULATED ATPASE IN THE COCCOLITH-PRODUCING COMPARTMENT OF CELLS OF PLEUROCHRYSIS SP. (PRYMNESIOPHYCEAE)1

Cell homogenates of Pleurochrysis sp. (CCMP299) were fractionated by means of sucrose gradients. Ca²⁺-stimulated ATPase (EC 3.6.1.3., ATP phosphohydrolase) was associated primarily with the plasma membrane, Golgi, and high density (1.21 g·cm^?3) membranous structures. Ca²⁺-stimulated ATPase was highly enriched in the latter. Based on treatments with Triton X-100 and NBD ceramide, we conclude that the high-density structures were membrane-delimited organelles. These vesicle-like organelles contained complex polysaccharides, a high concentration of calcium, and, upon microscopic examination, structures resembling coccoliths. These findings are consistent with observations on the known composition of coccoliths and the presumed mineralizing function of the sub-cellular coccolith-producing compartment. The high-density vesicles were linked to the Golgi by means of colchicine-sensitive materials, presumably microtubules. These data and prior ultrastructural observations by other investigators indicating vectorial assembly and secretion suggest that the subcellular movement of the newly formed coccoliths may be directed and/or powered by colchicine-sensitive cytoskeletal elements. We interpret the data to mean that the high-density vesicles represent the coccolith-producing compartment previously observed by others in electron micrographs.     

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.     

INDIAN OCEAN NANNOPLANKTON. II. HOLOCOCCOLITHOPHORIDS (CALYPTROSPHAERACEAE,PRYMNESIOPHYCEAE) WITH A REVIEW OF EXTANT GENERA1

Holococcolithophorids, pyrmnesiophytes having only one type of calcareous element in their coccoliths, are delicate and not commonly recorded in recent and fossil marine floras. There are few records of these organisms from the Indian Ocean and 26 species from there are included in this report. Although the group, generally assigned to a single family, the Calyptrosphaeraceae, may contain species that are part of the life history of heterococcolith-bearing cells in other stages, so little is known of this aspect of their biology that one must continue, at the present time, to treat them as independent taxonomic entities. A key is provided for the known holococcolithophorid genera. A new genus, Gliscolithus, and three new species, Gliscolithus amitakarenae, Calyptrosphaera heimdalae, and Helladosphaera pienaarii are described. The following new combinations are proposed: Calyptrolithina fragaria (Kamptner) comb. nov., Calyptrolithina gaarderae (Borsetti et Cati) comb. nov., Calyptrolithina isselii (Borsetti et Cati) comb. nov., Calyptrolithina lafourcadii (Lecal) comb. nov., Calyptrolithina magnaghii (Borsetti et Cati) comb. nov., Calyptrolithina multipora (Gaarder) comb. nov., Calyptrolithina porritectum (Heimdal) comb. nov., Calyptrolithina wettsteinii (Kamptner) comb. nov., Calyptrolithophora catillifera (Kamptner) comb. nov., Calyptrolithophora galea (Lecal-Schlauder) comb. nov., Dactylethra pirus (Kamptner) comb. nov., Helladosphaera arethusae (Kamptner) comb. nov., Helladosphaera gracilis (Kamptner) comb. nov., Homozygosphaera strigilis (Gaarder) comb. nov. and Syracolithus schilleri (Kamptner) comb. nov. The new combination in the genus Dactylethra Gartner for the first time brings an extant species into this genus that formerly contained only fossil species. It is pointed out that the enlarged zygoliths in stomatal areas of Corisphaera and the helladoliths in stomatal regions of Helladosphaera have intergrading types and are not distinctive enough characteristics to separate these genera.     

DIATOM PRODUCTIVITY COMPARED TO OTHER ALGAE IN NATURAL MARINE PHYTOPLANKTON ASSEMBLAGES1,2

Total productivity rates and the component attributed to the diatom proportion in the crop were measured in natural water samples taken from the Scripts Pier (La Jolla, California) at ca, monthly intervals in 1975–76. Two other measurements were done in water samples taken from Saanich Intel, Vancouver Islands, British Columbia (Canada). The percent of photosynthesis by diatoms was compared with the proportion of diatom carbon in the crop. In all instances, percent diatom photosynthesis was higher than percent diatom carbon for dialoms were higher than those of other algae. It is hypothesized that, in the sea, diatoms can outcompete other algae. However, the assemblages consist of some tens of species coexisting, and no portion of the crop out competes the others to exclusion.     

Coccolithophorid algae culture in closed photobioreactors

The feasibility of growth, calcium carbonate and lipid production of the coccolithophorid algae (Prymnesiophyceae), Pleurochrysis carterae, Emiliania huxleyi, and Gephyrocapsa oceanica, was investigated in plate, carboy, airlift, and tubular photobioreactors. The plate photobioreactor was the most promising closed cultivation system. All species could be grown in the carboy photobioreactor. However, P. carterae was the only species which grew in an airlift photobioreactor. Despite several attempts to grow these coccolithophorid species in the tubular photobioreactor (Biocoil), including modification of the airlift and sparger design, no net growth could be achieved. The shear produced by turbulence and bubble effects are the most likely reasons for this failure to grow in the Biocoil. The highest total dry weight, lipid and calcium carbonate productivities achieved by P. carterae in the plate photobioreactors were 0.54, 0.12, and 0.06 g L⁻¹ day⁻¹ respectively. Irrespective of the type of photobioreactor, the productivities were P. carterae > E. huxleyi > G. oceanica. Pleurochrysis carterae lipid (20–25% of dry weight) and calcium carbonate (11–12% of dry weight) contents were also the highest of all species tested. Biotechnol. Bioeng. 2011;108:2078–2087. © 2011 Wiley Periodicals, Inc.