Calcified structures and calcification in protists

Summary The diversity of calcified structures found in protists, the mechanisms utilized to form these structures, and the role these structures play in the taxonomy and systematics of the protists are presented. The two most frequently studied orders of protists which produce calcified structures, the coccolithophorids and foraminifera, are featured. However, consideration is given to the less known and least studied organisms.     

Physiological regulation of carbon fixation in the photosynthesis and calcification of coccolithophorids

Emiliania huxleyi and Gephyrocapsa oceanica are the predominant coccolithophorid species that produce blooms in the ocean and affect the global environment. These species are capable of carbon fixation by both photosynthesis for organic matter production and by intracellular calcification for coccolith production. Both processes were strongly affected by the nutrient status in a laboratory culture. The coccolith production was stimulated by the addition of a high concentration of sodium bicarbonate and by the depletion of phosphate. Interestingly, when the calcification was stimulated, the increase in cell number during algal growth was greatly suppressed and then the cell volume increased. When the growth rate was increased under nutrient-sufficient conditions, the cells became very small in size and most of them bore few or no coccoliths. The data from laboratory experiments show that the cell growth and calcification proceeded apparently independently at different phases. We, therefore, assume that the coccolithophorid blooms in the ocean might be separated into two phases; firstly, the increase in cell population might be triggered by an adequate supply of nutrients to enhance algal growth and then the calcification might subsequently be stimulated when the nutrients become depleted by substantial algal growth.     

Cytochemical and X-ray microanalysis studies of intracellular calcium pools in scale-bearing cells of the coccolithophoridEmiliania huxleyi

Summary Emiliania huxleyi is a coccolithophorid with a life cycle including a stage characterized by the occurrence of a scale-bearing cell type. The scales are composed of organic material and are produced in the cisternae of the Golgi apparatus. The present report deals with the ultrastructural calcium localization in scale-bearing cells using cation-precipitating agents. Cations were precipitated either with potassium pyroantimonate alone or according to a combined procedure in which cells are treated first with potassium oxalate, or potassium carbonate, or potassium phosphate, and then with potassium pyroantimonate. The distribution of electron-opaque deposits was the same when visualized by all four techniques. The most extensive deposits occurred in the Golgi apparatus, the peripheral space (a cellular compartment totally encompassing the protoplast), the multivesicular bodies, and the cell vacuole. X-ray microanalysis revealed that calcium was a constituent of the electron-opaque deposits. The uptake and transport of calcium, as universal functions of the Golgi apparatus, are discussed.     

The relationship between calcification and photosynthesis in the coccolithophorid Pleurochrysis carterae

Coccolithophorids are one of the dominant groups of marine phytoplankton. They are found in large numbers throughout the surface euphotic zone of the ocean, and are able to form large-scale blooms that persist for long periods of time. Coccolithophorid cells are covered by species-specific calcium carbonate crystals of various structures. In the process of calcification in coccolithophorids, Ca²⁺ is absorbed into cells from the culture medium, and a coccolith unit is formed inside the cell. Then, the coccolith unit extrudes to the cell surface where it is constructed into crystal layers. The formation of these crystals is regulated by cellular metabolism under different environmental conditions. The carbon biogeochemical cycle in the coccolithophorids involves both photosynthetic and calcification processes, which not only play an important role in population dynamics, but also in the global carbon cycle and climate change. However, one important question remains, namely, whether the relationship between photosynthesis and calcification is species-dependent. Previous studies have yielded controversial results, even in the same species. In this paper, we selected Pleurochrysis carterae, a coccolithophore species that frequently blooms in coastal areas, to study the relationship between calcification and photosynthesis. First, we studied population growth in a batch culture over several days. For batch cultures, P. carterae was inoculated into a 10 L bioreactor at an initial cell density of approximately 5 × 10⁴ cells mL^-1. The culture conditions were optimal for cell growth. Dissolved oxygen (DO) was detected during all the culture period, and the rate of photosynthetic oxygen evolution was calculated according the DO changes during the 12-h illumination period. Algal samples (10 mL) were collected during the population growth phases. The calcium carbonate content on the cell surface was determined each day by chemical titration. Next, we studied the relationship between photosynthesis and calcification at the cellular level by observing patterns of recalcification during a 12-h period. In this study, non-calcified cells were obtained by decalcifying calcified cells collected during the exponential growth period in MES-NaOH buffer solution (pH 5.5). The non-calcified cells were inoculated into culture media containing different concentrations of Ca²⁺ (0, 5, 20, 40, 50, or 100 mg L^-1). The rate of recalcification was determined by microscopic analyses in which the number of recalcified cells per 100 cells was counted at 0, 3, 6, 9, and 12 h of culture. Ca²⁺ absorbed into the cell was detected by measuring the fluorescence intensity of Fluo-3/AM labeled Ca²⁺. The rate of photosynthetic oxygen evolution in the non-calcified cell cultures was detected by measuring the changes in dissolved oxygen during the 12-h illumination period. The results showed that during the population growth period, the rate of photosynthetic oxygen evolution was inversely related to the calcium carbonate content per cell. When the amount of calcium carbonate on the cell surface increased, the relative photosynthetic ability (the rate of photosynthetic oxygen evolution) decreased, and vice versa. Both recalcification rates and photosynthetic oxygen evolution were affected by the extracellular calcium concentration. Non-calcified cells showed different recalcification abilities at different extracellular Ca²⁺ concentrations. The recalcification rate of non-calcified cells was positively correlated with the extracellular calcium concentration when [Ca²⁺] in the medium ranged from 0 to 100 mg L^-1. However, photosynthetic oxygen evolution was suppressed at higher cell calcification rates, especially when extracellular [Ca²⁺] was 50–100 mg L^-1. Our analyses of the population growth process and the cell recalcification process confirmed that photosynthesis is inversely related to calcification in P. carterae.     

Foraminifera and coccolithophorid assemblage changes in the Panama Basin during the last deglaciation: Response to sea-surface productivity induced by a transient climate change

The responses of community assemblages of planktonic and benthonic foraminifera and coccolithophorids to transient climate change are explored for the uppermost 2 m of cores ODP677B (1.2°N; 83.74°W, 3461 m) and TR163-38 (1.34°S; 81.58°W, 2200 m), for the last ∼ 40 ka. Results suggest that the deglaciation interval was a time of increased productivity and a major reorganization of planktonic trophic webs. The succession in dominance between the planktonic foraminifera species Globorotalia inflata, Globigerina bulloides, and Neogloboquadrina pachyderma denote four periods of oceanographic change: (1) advection (24-20 ka), (2) strong upwelling (20-15 ka), (3) weak upwelling (14-8 ka) and (4) oligotrophy (8 ka to present). Strong upwelling for the deglaciation interval is supported by the low Florisphaera profunda/other coccolithophorids ratio and the high percentage abundance of Gephyrocapsa oceanica. Benthonic foraminifera assemblage changes are different in both cores and suggest significant regional variations in surface productivity and/or oxygen content at the seafloor, and a decoupling between surface productivity and export production to the seafloor. This decoupling is evidenced by the inverse relationship between the percentage abundance of infaunal benthonic foraminifera and the percentage abundance of N. pachyderma. The terrigenous input of the Colombian Pacific rivers, particularly the San Juan River, is suggested as a possible mechanism. Finally, the Globorotalia cultrata/Neogloboquadrina dutertrei ratio is used to reconstruct the past influence of the Costa Rica Dome-Panama Bight and cold tongue upwelling systems in the Panama Basin. A northern influence is suggested for the late Holocene (after 5 ka) and the last glacial (before 20 ka), whereas a southern influence is suggested for the 20-5 ka interval. There is a correspondence between our reconstructed northern and southern influences and previously proposed positions of the Intertropical Convergence Zone (ITCZ) after the Last Glacial Maximum (LGM).     

Polyanion-mediated mineralization — assembly and reorganization of acidic polysaccharides in the Golgi system of a coccolithophorid alga during mineral deposition

Summary Immunolocalization of two highly acidic polysaccharides (PS-1 and PS-2) in a calcifying algaPleurochrysis carterae is described throughout the mineralization process, from before crystal nucleation through the cessation of crystal growth. This unicellular coccolithophorid alga is a useful model for mineralization because it produces calcified scales known as coccoliths in homogeneous cell culture. PS-1 and PS-2 were localized in the crystal coats of mature coccoliths and in electron dense Golgi particles. The polyanions are synthesized in medial Golgi cisternae and co-aggregate with calcium ions into discrete 25 nm particles. Particle-laden vesicles bud from cisternal margins and fuse with a coccolith-forming saccule containing an organic oval-shaped scale which forms the base of the future coccolith. The particles are localized on the base before the onset of mineral deposition and are present in the coccolith saccule throughout the period of crystal (CaCO₃) nucleation and growth. During the final phase of coccolith formation, the particles disappear, and the mature crystals acquire an amorphous coat containing PS-1 and PS-2 polysaccharides which remain with the mineral phase after the coccoliths are extruded from the cell. Postulated mechanisms of polyanion-mediated mineralization are reviewed and their relevance to the calcification of coccoliths is addressed.Abbreviations PS-1 polysaccharide one - PS-2 polysaccharide two - BSA bovine serum albumin - SDS sodium dodecyl sulfate - MES 2-(N-morpholino)-ethanesulfonic acid - EDTA ethylenediaminetetraacetic acid - DHA 3-deoxy-lyxo-2-heptulosaric acid - TCA trichloroacetic acid