A NEW CELL STAGE IN THE HAPLOID‐DIPLOID LIFE CYCLE OF THE COLONY‐FORMING HAPTOPHYTE PHAEOCYSTIS ANTARCTICA AND ITS ECOLOGICAL IMPLICATIONS1

Few members of the well‐studied marine phytoplankton taxa have such a complex and polymorphic life cycle as the genus Phaeocystis. However, despite the ecological and biogeochemical importance of Phaeocystis blooms, the life cycle of the major bloom‐forming species of this genus remains illusive and poorly resolved. At least six different life stages and up to 15 different functional components of the life cycle have been proposed. Our culture and field observations indicate that there is a previously unrecognized stage in the life cycle of P. antarctica G. Karst. This stage comprises nonmotile cells that range in size from ～4.2 to 9.8 μm in diameter and form aggregates in which interstitial spaces between cells are small or absent. The aggregates (hereafter called attached aggregates, AAs) adhere to available surfaces. In field samples, small AAs, surrounded by a colony skin, adopt an epiphytic lifestyle and adhere in most cases to setae or spines of diatoms. These AAs, either directly or via other life stages, produce the colonial life stage. Culture studies indicate that bloom‐forming, colonial stages release flagellates (microzoospores) that fuse and form AAs, which can proliferate on the bottom of culture vessels and can eventually reform free‐floating colonies. We propose that these AAs are a new stage in the life cycle of P. antarctica, which we believe to be the zygote, thus documenting sexual reproduction in this species for the first time.     

COLONY SIZE OF PHAEOCYSTIS ANTARCTICA (PRYMNESIOPHYCEAE) AS INFLUENCED BY ZOOPLANKTON GRAZERS1

The haptophyte Phaeocystis antarctica G. Karst. is a dominant phytoplankton species in the Ross Sea, Antarctica, and exists as solitary cells and mucilaginous colonies that differ by several orders of magnitude in size. Recent studies with Phaeocystis globosa suggest that colony formation and enlargement are defense mechanisms against small grazers. To test if a similar grazer‐induced morphological response exists in P. antarctica, we conducted incubation experiments during the austral summer using natural P. antarctica and zooplankton assemblages. Dialysis bags that allowed exchange of dissolved chemicals were used to separate P. antarctica and zooplankton during incubations. Geometric mean colony size decreased by 35% in the control, but increased by 30% in the presence of grazers (even without physical contact) over the 15 d incubation. The estimated colonial‐to‐solitary cell carbon ratio was significantly higher in the grazing treatment. These results suggest that P. antarctica colonies would grow larger in the presence of indigenous zooplankton and skew the carbon partitioning significantly toward the colonial phase. While these observations show that the colony size of P. antarctica was affected by a chemical signal related to grazers, the detailed nature and ecological significance of this signal remain unknown.     

MORPHOLOGICAL AND GENETIC CHARACTERIZATION OF PHAEOCYSTIS CORDATA AND P. JAHNII (PRYMNESIOPHYCEAE), TWO NEW SPECIES FROM THE MEDITERRANEAN SEA

Two new Phaeocystis species recently discovered in the Mediterranean Sea are described using light and electron microscopy, and their systematic position is discussed on the basis of an analysis of their nuclear-encoded small-subunit ribosomal RNA gene (SSU rRNA) sequences. Phaeocystis cordata Zingone et Chrétiennot-Dinet was observed only as flagellated unicells. Cells are heart shaped, with two flagella of slightly unequal length and a short haptonema. The cell body is covered with two layers of thin scales. The outermost layer scales are oval, with a faint radiating pattern, a raised rim, and a modest central knob. The inner-layer scales are smaller and have a faint radiate pattern and an inflexed rim. Cells swim with their flagella close together, obscuring the haptonema, pushing the cell, and causing it to rotate about its longitudinal axis while moving forward. Phaeocystis jahnii Zingone was isolated as a nonmotile colony. It forms loose aggregates of cells embedded in a mucilaginous, presumably polysaccharide matrix without a definite shape or visible external envelope. The flagellated stage has the features typical of other Phaeocystis species. Cells are rounded in shape and slightly larger than P. cordata. The cell body is covered with extremely thin scales of two different sizes with a very faint radiating pattern toward their margin. Swimming behavior is similar to that of P. cordata, with the flagella in a posterior position as the cells swim. The SSU rRNA sequence analysis indicated that both species are distinct from other cultivated Phaeocystis species sequenced to date. Regions previously identified as specific for the genus Phaeocystis are not found in P. jahnii, and new genus-specific regions have been identified. P. cordata is more closely related to the colonial species P. globosa, P. antarctica, and P. pouchetii and has branched prior to the divergence of the warm-water P. globosa species complex from the cold-water species P. antarctica and P. pouchetii. These results are discussed within a framework ofthe available data on the evolution of the world’s oceans.     

THE EFFECT OF IRON LIMITATION ON THE PHOTOPHYSIOLOGY OF PHAEOCYSTIS ANTARCTICA (PRYMNESIOPHYCEAE) AND FRAGILARIOPSIS CYLINDRUS (BACILLARIOPHYCEAE) UNDER DYNAMIC IRRADIANCE1

The effects of iron limitation on photoacclimation to dynamic irradiance were studied in Phaeocystis antarctica G. Karst. and Fragilariopsis cylindrus (Grunow) W. Krieg. in terms of growth rate, photosynthetic parameters, pigment composition, and fluorescence characteristics. Under dynamic light conditions mimicking vertical mixing below the euphotic zone, P. antarctica displayed higher growth rates than F. cylindrus both under iron (Fe)–replete and Fe‐limiting conditions. Both species showed xanthophyll de‐epoxidation that was accompanied by low levels of nonphotochemical quenching (NPQ) during the irradiance maximum of the light cycle. The potential for NPQ at light levels corresponding to full sunlight was substantial in both species and increased under Fe limitation in F. cylindrus. Although the decline in F_v/F_m under Fe limitation was similar in both species, the accompanying decrease in the maximum rate of photosynthesis and growth rate was much stronger in F. cylindrus. Analysis of the electron transport rates through PSII and on to carbon (C) fixation revealed a large potential for photoprotective cyclic electron transport (CET) in F. cylindrus, particularly under Fe limitation. Probably, CET aided the photoprotection in F. cylindrus, but it also reduced photosynthetic efficiency at higher light intensities. P. antarctica, on the other hand, was able to efficiently use electrons flowing through PSII for C fixation at all light levels, particularly under Fe limitation. Thus, Fe limitation enhanced the photophysiological differences between P. antarctica and diatoms, supporting field observations where P. antarctica is found to dominate deeply mixed water columns, whereas diatoms dominate shallower mixed layers.     

MORPHOLOGY,PLOIDY, PIGMENT COMPOSITION,AND GENOME SIZE OF CULTURED STRAINS OF PHAEOCYSTIS (PRYMNESIOPHYCEAE)1

We examined cell morphology, ploidy level, cell size, pigment composition, and genome size in 16 cultured strains of Phaeocystis Lagerheim. Two strains originated from the Antarctic, 3 from the tropical Western Atlantic, and 11 from temperate regions (Eastern Atlantic, English Channel, North Sea, and Mediterranean Sea). Thirteen strains made colonies morphologically similar to P. glo-bosa Scherffel, whereas three never formed colonies under any circumstances. Five-rayed star-like structures with filaments were observed in 11 strains. In several strains, two ploidy levels were observed, one (haploid) linked to flagellates and one (diploid) linked to colonies. Cell size did not appear to be a very good criterion for distinguishing strains since size distributions overlapped. Pigment analysis by reversed-phase-high-performance liquid chroma-tography allowed the strains to be grouped into three clusters that differed from each other mainly by the relative proportions of three carotenoids: fucoxanthin, 19′-hex-anoyloxyfucoxanthin, and diadinoxanthin. All strains contained low levels of 19′-butanoyloxyfucoxanthin. Differences in genome size measured by flow cytometry delimited at least five groups. On the basis of both pigment composition and genome size, six clusters were defined, one corresponding to an Antarctic species (possibly P. antarc-tica), one to P. globosa, and the rest probably to several yet-undescribed species or subspecies. Two main conclusions emerge from this study. First, the taxonomy of the genus Phaeocystis needs to be clarified through a combination of morphological, biochemical, and molecular studies. Second, sexuality is a prevalent phenomenon in Phaeocystis, but controls of the sexual cycle are most likely strain-dependent.     

Differentiation betweenPhaeocystis pouchetii (Har.) Lagerheim andPhaeocystis globosa Scherffel

An Arctic clone ofPhaeocystis pouchetii LAGERHEIM was compared toPhaeocystis globosa SCHERFFEL isolated from the southern North Sea with regard to temperature tolerance and colony shapes. Already youngP.pouchetii colonies (<100 m) show the typical distribution of the cells in groups, separated from each other by wide zones of cell-free mucilage; the maximum colony size is ca 2 mm in diameter.P.pouchetii colonies form clouds with bubble-like vesicles, spherical colony-shapes are seldom found.P.globosa colonies are spherical up to a size of 2 mm; the cells are distributed homogeneously over the periphery of the colonies. A pouchetii-like distribution of cells never occurs either in the spherical young colonies or in the pear-shaped old colonies (size up to 8 mm). A development from the colony shape of the globosa-type to the pouchetii-type or vice versa was never found. Therefore the colony shape has to be considered a constant distinctive character. Single cells ofP.pouchetii andP.globosa cannot be separated from each other by using the light microscope; this also holds for the flagellates and the non-motile cells.P.pouchetii grows well between 0°C and 14°C,P.globosa between 4°C and 22°C, respectively. Because of the distinctive differences in the morphology of the colonies and the differences in temperature tolerances we propose thatPhaeocystis globosa should no longer be considered conspecific withPhaeocystis pouchetii.     

棕囊藻属(Phaeocystis)的分类与生活史(综述)

棕囊藻属Phaeocystis（定鞭藻纲Prymnesiophyceae)的分类问题目前还有争论。其种的分类标准是以初始的群体形态、地理分布、细胞特征等以及分子生物学特征,如染色体倍性,基因组大小等为依据。基于以上各种分类特征,目前比较确定的棕囊藻属藻类有四种：一种是只观察到单细胞形态的凹孔棕囊藻（P.scrobiculata),另外三种是能够形成群体的波切棕囊藻（P.pouchetii)、球形棕囊藻（P.globosa)和南极棕囊藻（P.antarctica)。棕囊藻具有一个复杂的异形生活史,介于几种游离的单细胞（不动的细胞,具有鞭毛的动细胞,小游动孢子以及可能存在的大游动孢子）和群体之间的形态交替。但其生活史中仍有许多不确定的问题。     

ISOLATION AND CHARACTERIZATION OF A VIRUS INFECTING PHAEOCYSTIS POUCHETII (PRYMNESIOPHYCEAE)1

A virus infecting the haptophyte Phaeocystis pouchetii (Hariot) Lagerheim was isolated from Norwegian coastal waters in May 1995 at the end of a bloom of this phytoplankter. The virus was specific for P. pouchetii because it did not lyse 10 strains of P. globosa Scherffel, Phaeocystis sp., and P. antarctica Karsten. It was a double-stranded DNA virus, and the viral particle was a polyhedron with a diameter of 130–160 nm. The virus had a main polypeptide of about 59 kDa and at least five minor polypeptides between 30 and 50 kDa. The latent period of the virus when propagated in cultures of P. pouchetii was 12–18 h, and the time required for complete lysis of the cultures was about 48 h. The burst size was estimated to be 350–600 viral particles per lysed cell.     

STRATEGIES AND RATES OF PHOTOACCLIMATION IN TWO MAJOR SOUTHERN OCEAN PHYTOPLANKTON TAXA: PHAEOCYSTIS ANTARCTICA (HAPTOPHYTA) AND FRAGILARIOPSIS CYLINDRUS (BACILLARIOPHYCEAE)1

We investigated rates and mechanisms of photoacclimation in cultures of Phaeocystis antarctica G. Karst. and Fragilariopsis cylindrus (Grunow) Willi Krieg, phytoplankton taxa that each dominate distinct areas of the Ross Sea, Antarctica. Both P. antarctica and F. cylindrus acclimated to increases in irradiance by reducing the effective size of the pigment antenna (σPSII) via xanthophyll‐cycle activity and reductions in chl. While enhanced photoprotection facilitated increases in specific growth rate and eventually led to higher light‐saturated photosynthetic rates (P^cell_m) in P. antarctica, increases in those variables were much smaller in F. cylindrus. In response to a lower irradiance, relaxation of xanthophyll‐cycle activity led to an increase in σPSII in both taxa, which occurred much more slowly in F. cylindrus. A surprising increase in specific growth rate over the first 36 h of acclimation in P. antarctica may have facilitated the significant reductions in P^cell_m observed in that taxon. In general, P. antarctica acclimated more quickly to changes in irradiance than F. cylindrus, exhibited a wider range in photosynthetic rates, but was more susceptible to photoinhibition. This acclimation strategy is consistent with growth in deeply mixed water columns with variations in irradiance that allow time for repair. In contrast, the slower acclimation rates, extensive photoprotection, and low photoinhibition exhibited by F. cylindrus suggest that it does not require the same period for repair as P. antarctica and is best suited for growth in habitats with relatively uniform irradiance, such as shallow mixed layers or sea ice.     

The vitamin B requirement of Phaeocystis globosa (Prymnesiophyceae)

In batch cultures of flagellates and non-flagellate cells ofPhaeocystis globosa, the biomass yield was significantly enhancedby the addition of a mixture of the vitamins thiamine (B1),cyanocobalamin (B12) and biotin (H). A bioassay with B1 andB12 using the non-flagellate cells of P.globosa showed thatthis prymnesiophyte is a B1 auxotroph. The bioassay also indicateda significant difference in growth rate between culture mediumwith 10 nmol l^–1 B1 (µ = 0.80 day^–1) and culturemedium with 10 nmol l^–1 B12 (µ = 0.52 day^–1).These findings are discussed in relation to the hypothesis thatcentric diatoms, through vitamin B1 excretion or B12 depletion,initiate Phaeocystis blooms. It is concluded, however, thatan alternative hypothesis, that diatoms provide a solid substratefor colony initiation, has more experimental support.     

摄食信息对球形棕囊藻和布氏双尾藻竞争结果的影响

浮游动物的摄食信息能增大棕囊藻囊体体积,囊体形成被认为是棕囊藻的诱导性防御机制。利用桡足类火腿伪镖水蚤和异养甲藻海洋尖尾藻释放的摄食信息,研究了诱导性防御对球形棕囊藻和布氏双尾藻的竞争的影响。结果表明,球形棕囊藻接收了火腿伪镖水蚤和海洋尖尾藻释放的摄食信息之后形成更大的囊体。防御启动后的球形棕囊藻比未接收摄食信息的球形棕囊藻更快地形成囊体,且囊体维持的时间更长。对照组和火腿伪镖水蚤摄食信息诱导的球形棕囊藻的生物体积比布氏双尾藻更高,且球形棕囊藻在竞争中占优势;而海洋尖尾藻摄食信息诱导的球形棕囊藻生物体积低于布氏双尾藻,且球形棕囊藻相对布氏双尾藻的竞争力下降。微型浮游动物海洋尖尾藻摄食信息导致球形棕囊藻相对硅藻布氏双尾藻的竞争力的下降,有利于解释硅藻先于棕囊藻发生藻华。     

PHOTOPHYSIOLOGY IN TWO SOUTHERN OCEAN PHYTOPLANKTON TAXA: PHOTOSYNTHESIS OF PHAEOCYSTIS ANTARCTICA (PRYMNESIOPHYCEAE) AND FRAGILARIOPSIS CYLINDRUS (BACILLARIOPHYCEAE) UNDER SIMULATED MIXED‐LAYER IRRADIANCE1

In the Ross Sea, the prymnesiophyte Phaeocystis antarctica G. Karst. dominates deeply mixed water columns, while diatoms dominate shallower mixed layers. Understanding what controls the dynamics of these two phytoplankton taxa is essential because they dominate virtually all coastal polar waters, have different nutrient utilization characteristics, and support dissimilar food webs. We cultured two strains of P. antarctica and one strain of the diatom Fragilariopsis cylindrus (Grunow) Willi Krieg under three dynamic irradiance regimes that simulated different mixed‐layer depths and measured their photosynthetic characteristics, cellular pigment concentrations, and cellular carbon and nitrogen content. In both species, chl a–normalized maximum carbon uptake rate (P_m^* ) and specific growth rate were highest in the deeply mixed treatment that had a dark period. In all irradiance treatments, both (P_m^* ) and photosynthetic efficiency (α*) were greater for the two P. antarctica strains than for the F. cylindrus strain. In contrast, P. antarctica strains were more susceptible to photoinhibition (β*) than the F. cylindrus strain. When photosynthetic rates of each phytoplankton taxon were normalized by cellular particulate organic carbon (POC), the difference in the maximal photosynthetic rate () was generally reduced. In the dynamic irradiance treatment that simulated the shallowest mixed‐layer irradiance, all three phytoplankton had similar ; however, the diatom had a 2‐fold higher POC‐normalized photosynthetic efficiency (α^C). Finally, we performed calculations using the measured POC‐normalized photosynthetic parameters to show that α^C and can play a greater role than β^C in determining the competitive outcome between P. antarctica and F. cylindrus in both shallow and deep mixed‐layer environments of the Ross Sea.     

COMPARISONS OF NITRATE UPTAKE, STORAGE, AND REDUCTION IN MARINE DIATOMS AND FLAGELLATES

Diatoms, but not flagellates, have been shown to increase rates of nitrogen release after a shift from a low growth irradiance to a much higher experimental irradiance. We compared NO₃ ^? uptake kinetics, internal inorganic nitrogen storage, and the temperature dependence of the NO₃ ^? reduction enzymes, nitrate (NR) and nitrite reductase (NiR), in nitrogen‐replete cultures of 3 diatoms (Chaetoceros sp., Skeletonema costatum, Thalassiosira weissflogii) and 3 flagellates (Dunaliella tertiolecta, Pavlova lutheri, Prorocentrum minimum) to provide insight into the differences in nitrogen release patterns observed between these species. At NO₃ ^? concentrations <40 μmol‐N·L ^{? 1}, all the diatom species and the dinoflagellate P. minimum exhibited saturating kinetics, whereas the other flagellates, D. tertiolecta and P. lutheri, did not saturate, leading to very high estimated K _s values. Above ～60 μmol‐N·L ^{? 1}, NO₃ ^? uptake rates of all species tested continued to increase in a linear fashion. Rates of NO₃ ^? uptake at 40 μmol‐N·L ^{? 1}, normalized to cellular nitrogen, carbon, cell number, and surface area, were generally greater for diatoms than flagellates. Diatoms stored significant amounts of NO₃ ^? internally, whereas the flagellate species stored significant amounts of NH₄ ⁺ . Half‐saturation concentrations for NR and NiR were similar between all species, but diatoms had significantly lower temperature optima for NR and NiR than did the flagellates tested in most cases. Relative to calculated biosynthetic demands, diatoms were found to have greater NO₃ ^? uptake and NO₃ ^? reduction rates than flagellates. This enhanced capacity for NO₃ ^? uptake and reduction along with the lower optimum temperature for enzyme activity could explain differences in nitrogen release patterns between diatoms and flagellates after an increase in irradiance.     

THE LIFE CYCLE OF PEDIASTRUM SIMPLEX1

A sexual cycle and a newly discovered mode of asexual reproduction by azygotes arc described for Pediastrum simplex. In the sexual cycle isogametes from colony cells fuse in pairs, develop into zygotes, which grow and divide, producing motile cells which become angular polyeders. Polyeders may divide internally to form 4-celled colonies, or they enlarge, cleave, and produce zoospores or nonmotile cells. Zoospores are released in vesicles, swarm, adhere to each other, and form, flat colonies I cell thick. The nonmotile cells adhere to each other, enlarge, break out of the polyeder, and become spheroidal colonies. In the asexual cycle, nonfusing gametes, and motile cells derived from colony cells, become 4-spined azygotes which enlarge, divide internally, and release motile cells developing into polyeders. These polyeders develop further like those from zygotes.     

Lotharella amoeboformis sp. nov.: A new species of chlorarachniophytes from Japan

The ultrastructure, morphology and life cycle of a new chlorarachniophyte alga collected from Okinawa in Japan have been studied. The life cycle of this alga consists of amoeboid, wall‐less round, coccoid and flagellated cells in culture condition; however, the coccoid and flagellate cells are very rare. The pyrenoid ultra‐structure of this alga is the same as that of a previously described species, Lotharella globosa. Since pyrenoid ultrastructure has been adopted as the main criterion for the generic classification of the chlorarachniophytes, the present alga is placed in Lotharella. However, the present alga has a dominant amoeboid cell stage and a reduced walled‐cell stage in the life cycle, while in L. globosa, the walled‐cell stage is dominant and there is no amoeboid cell stage. Therefore the present alga is described as a new species of Lotharella: Lotharella amoeboformis Ishida et Y. Hara sp. nov.