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.     

Low Temperature Stimulates Cell Enlargement and Intracellular Calcification of Coccolithophorids

Temperature effect on growth, cell size, calcium uptake activity, coccolith production was studied in coccolith-producing haptophytes, Emiliania huxleyi (Lohmann) Hay & Mohler (strain EH2) and Gephyrocapsa oceanica Kamptner (strain GO1) (Coccolithophorales, Prymnesiophyceae). E. huxleyi grew at a wider temperature range (10°–25°C), while G. oceanica growth was limited to warmer temperatures (20°–25°C). Cell size was inversely correlated with temperature. At low temperature, the enlargement of chloroplasts and cells and the stimulation of coccolith production were morphologically confirmed under fluorescent and polarization microscopes, respectively. ⁴⁵ Ca uptake by E. huxleyi at 10°C was greatly increased after a 5-day lag and exceeded that at 20°C. These results clearly showed that low temperature suppressed coccolithophorid growth but induced cell enlargement and as stimulated the intracellular calcification that produces coccoliths.     

ISOLATION AND CHARACTERIZATION OF GOLGI FROM COCCOLITHUS PELAGICUS (PRYMNESIOPHYCEAE)1

Cells of the marine alga Coccolithus pelagicus (Wal-lich)J. Schiller grown in axenic cultures were homogenized and fractionated. The distribution of organelle markers was assessed enzymatically after centrifugation through zonal, density, and flotation gradients made with sucrose, sorbitol, or Percoll. Mitochondria (1.19 g·cm^-3) and chloroplasts (1.15 g·cm^-3) were recovered in sucrose gradients at densities similar to those observed for higher plants and most algae. The position of endoplasmic reticulum and plasma membrane in the gradients was monitored by NADPH cytochrome c reductase and vanadate-sensitive Mg²⁺-ATPase, respectively. Higher plant Golgi markers, latent undine diphosphatase (UDPase) and glucan synthase I, were colocalized at a density range including two peaks of activity at 1.13–1.15 g·cm^-3. Bound calcium was associated with high density (1.15 g·cm^-3) membranes. Ca²⁺-stimulated ATPase was found at high levels on membranes that did not coisolate with the latent UDPase-containing membranes. The Ca²⁺-stimulated ATPase, a possible participant during calcification, was associated with a chloroplast-enriched fraction in all the organelle separation systems. However, about 30% of the total activity was separated from both the chloroplasts and Golgi on 0–70% Percoll gradients containing 0.4 M sucrose. The possible relationship of the Golgi and the high-density organelle exhibiting Ca²⁺-stimulated ATPase to coccolithogenesis and the process of calcification and crystal formation is discussed.     

HETEROMORPHIC LIFE HISTORIES IN ARCTIC COCCOLITHOPHORIDS (PRYMNESIOPHYCEAE)1

During three visits to Disko Bay, West Greenland, we found four different types of prymnesiophyte flagellates with heterococcoliths from species of the genera Papposphaera Tangen and Pappomonas Manton and Oates in characteristic and consistent combinations with holococcoliths from species of the genera Turrisphaera Manton, Sutherland, and Oates and Trigonaspis Thomsen. We conclude that several taxa previously considered to be autonomous species are, in fact, part of life histories combining hetero- and holococcolithophorid forms in a manner somewhat similar to that known from studies of cultured strains of Coccolithus pelagicus (Wallich) Schiller and Crystallolithus hyalinus (Gaarder) Markali. Similarly, Calciarcus Manton, Sutherland and Oates and Wigwamma Manton, Sutherland and Oates also may be alternate phases of a coccolithophorid life history.     

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.     

MOLECULAR PHYLOGENY OF THE HAPTOPHYTA BASED ON THE rbcL GENE AND SEQUENCE VARIATION IN THE SPACER REGION OF THE RUBISCO OPERON

A phylogeny of 21 haptophyte algae was inferred by maximum parsimony, neighbor-joining, and maximum likelihood analyses of sequences of the plastid-encoded gene, rbcL. Sequence variation in the spacer region of the RUBISCO operon was also investigated. In all the rbcL trees constructed, the haptophytes form two distinct clades: one includes the Pavlovales and the other includes the Prymnesiales, Coccosphaerales, and Isochrysidales (all sensu Parke and Green 1976 . This relationship coincides with the recent taxonomic treatment splitting the division into two subclasses, the Prymnesidae and Pavlovidae ( Cavalier-Smith 1989 ) or the Prymnesiophycidae and the Pavlovophycidae using botanical suffixes ( Jordan and Green 1994 ), or into two classes, the Patelliferea and the Pavlovea ( Cavalier-Smith 1993 ). In the Prymnesiophycidae, all the coccolithophorids examined are placed in a single clade, which suggests a single origin of the coccolithophorids and the ability of coccolith formation in the haptophytes. The genus Chrysochromulina is polyphyletic. Species of Chrysochromulina with a very long haptonema and a compressed cell body (typical of species including the type C. parva Lackey) form a clade, including Imantonia, that is often classified in the Isochrysidales in the neighbor-joining tree, whereas some species possessing a nontypical cell body and cell covering form a clade with Prymnesium and Platychrysis in all trees. It is suggested that loss of the haptonema in Imantonia and the reduction in Prymnesium and Platychrysis occurred secondarily and independently in two different lineages. Within the coccolithophorids, four clades are recognized: Pleurochrysis, Calyptrosphaera-Cruciplacolithus-Calcidiscus-Umbilicosphaera, Helicosphaera, and Emiliania-Gephyrocapsa. A non-coccolith-bearing haptophyte, Isochrysis, is an ingroup of the Emiliania-Gephyrocapsa clade, suggesting its secondary loss of the ability to form a coccolith. Sequence comparison of the spacer region of RUBISCO operon supports most results obtained in the analysis of rbcL sequences. Monophyly of the Prymnesiales sensu Parke and Green is still unclear because of low (<50%) bootstrap support for this group.     

THE OCCURRENCE OF CHLOROPHYLL C1 AND C2 IN ALGAE1

Forty-two species of chlorophyll c-containing algae (diatoms, dinoflagellates, chrysomonads, haptophytes, cryptomonads and xanthophytes) were examined for their content of chlorophyll c₁ and c₂. This work, and recent studies on c₁/c₂ distribution in the literature (total 86 species), show that chlorophyll c₂ is universal to all algae examined. Chlorophyll c₁ occurs in addition to c₂ in brown seaweeds, diatoms, chrysomonads, haptophytes (coccolithophorids) xanthophytes and the, fucoxanthin-containing dinoflagellates; c₂ only occurs in dinoflagellates and cryptomonads. Two exceptions to the generalizations are one dinoflagellate and one cryptomonad containing c₁ in addition to c₂ No explanation can be offered on present knowledge for these exceptions. No alga was found containing only chlorophyll c₁. Chlorophyll c, far from being a minor accessory chlorophyll, occurred in amounts almost equal to chlorophyll a(some diatoms and dinoflagellates) or ranged from 50 to 20% of the chlorophyll a (diatoms, dinoflagellates, chrysomonads, cryptomonads, browns). Xanthophytes, however, contained only trace amounts of chlorophyll c with ratios of chlorophyll a:c ranging from 55:1 to 116:1 on a weight basis. In those algae with both chlorophyll c components, c₁ and c₂, occurred either in equal amounts, or chlorophyll c₂was twice the c₁ content.     

Polymorphic microsatellite loci in global populations of the marine coccolithophorid Emiliania huxleyi

The marine coccolithophorid Emiliania huxleyi is an important component of the marine carbon cycle because bloom development results in the export of calcium carbonate from the ocean surface to the abyss. Laboratory and field studies demonstrate significant biogeographical, ecological, physiological and morphological plasticity in E. huxleyi and suggest high underlying genetic variability. Here we describe seven polymorphic microsatellite loci from the E. huxleyi genome and their degree of polymorphism in clonal isolates of different geographical origin. Our results indicate a high degree of genetic diversity within E. huxleyi.