Living in a Phaeocystis colony: a way to be a successful algal species

Evidence is provided showing that in two species of Phaeocystis (P. globosa and P. pouchetii) the colonial cells possess a much higher growth rate than the single cells when grown under identical conditions. Based on the DNA-cell-cycle method gross growth rate of colony cells exceeded those of co-occurring single cells by a factor 1.5 up to 3.8. The dominance of colonies in blooms of Phaeocystis can therefore be primarily due to their significantly high growth rate allowing a rapid bloom formation.Both Phaeocystis species showed ultradian growth but differed in timing of the initiation of the second DNA replication phase. In both species the first DNA-replication period started at the end of the (local) light period and was completed in the early dark period. In P. globosa this was immediately followed by the second DNA-replication period (first half of the dark period). In P. pouchetii this process was delayed by ca. 12 h until the middle of the light period (local noon).Flow cytometric analysis of the cell size and chlorophyll fluorescence showed little variation in colony and single cells of P. pouchetii. In contrast, colonies of P. globosa showed often the presence of two cell morphs, co-occurring in the same colony. The size of both morphs was identical but they differed in chlorophyll fluorescence up to a factor 4. In general the high chlorophyll cell morph dominated (>70% of the total colony cells). Both colony cell morphs were observed in cultures, mesocosms differing in N/P ratio but also in the field.     

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.     

Selective grazing of Temora longicornis in different stages of a Phaeocystis globosa bloom—a mesocosm study

Selective grazing of a calanoid copepod Temora longicornis was measured during different stages of a Phaeocystis globosa bloom, in order to reveal (1) if T. longicornis feeds on single cells and/or colonies of P. globosa in the presence of alternative food sources, (2) if copepod food selection changes during the initiation, maintenance, collapse and decay of a P. globosa bloom and (3) if P. globosa dominated food assemblage provides a good diet for copepod egg production. Our results show low but constant feeding on small colonies of P. globosa, irrespective of the type or concentration of alternative food sources. In contrast, feeding on single cells was never significant, and the total contribution of P. globosa to carbon ingestion of T. longicornis was minor. T. longicornis fed most actively on the decaying colonies, whereas during the peak of the bloom copepods selected against P. globosa. Mostly, T. longicornis fed unselectively on different food particles: before the bloom, the major part of the diet consisted of diatoms, whereas during and after the bloom copepod diet was dominated by dinoflagellates and ciliates. Egg production was highest during the decay of the bloom, coinciding with highest proportional ingestion of heterotrophic organisms, but was not seriously reduced even during the peak of the bloom. We conclude that P. globosa blooms should not threaten survival of copepod populations, but the population recruitment may depend on the type (and concentration) of the dominant heterotrophs present during the blooms. Due to relatively unselective grazing, the impact of T. longicornis to the initiation of a Phaeocystis bloom is considered small, although grazing on decaying colonies may contribute to the faster termination of a bloom.     

An examination of the role of colonial <Emphasis Type="Italic">Phaeocystis antarctica</Emphasis> in the microbial food web of the Ross Sea

The extensive buildup of phytoplankton biomass in the Ross Sea conflicts with the view that high rates of herbivory occur in all regions of the Southern Ocean. Nano and microplanktonic consumers comprise a significant fraction of total plankton biomass; however, the importance of grazing remains uncertain in the Ross Sea. Microzooplankton ingestion of solitary and colonial cells of Phaeocystis antarctica were calculated using a novel live-staining fluorescently-labeled algae method. Different morphotypes of P. antarctica were stained different colors, mixed, and observed inside Euplotes to determine their feeding preference. The blue (7-aminocoumarin) (CMAC) stain was used on the colonies and the green (CMFDA) CellTracker Probe was used on solitary cells. Both morphotypes can be seen inside the food vacuoles of the ciliate, supporting the idea that microzooplankton are capable of ingesting cells within the colonial matrix. This suggests that P. antarctica colonies enter the microbial loop in the Ross Sea before sedimentation.     

Molecular genetic delineation of Phaeocystis species (Prymnesiophyceae) using coding and non-coding regions of nuclear and plastid genomes

Sequence variation among 22 isolates representing a global distribution of the prymnesiophyte genus Phaeocystis has been compared using nuclear-encoded 18S rRNA genes and two non-coding regions: the ribosomal DNA internal transcribed spacer 1 (ITS1) separating the 18S rRNA and 5.8S rRNA genes and the plastid ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO) spacer flanked by short stretches of the adjacent large and small subunits (rbcL and rbcS). 18S rRNA can only resolve major species complexes. The analysis suggests that an undescribed unicellular Phaeocystis sp. (isolate PLY 559) is a sister taxon to the Mediterranean unicellular Phaeocystis jahnii; this clade branched prior to the divergence of all other Phaeocystis species, including the colonial ones. Little divergence was seen among the multiple isolates sequenced from each colonial species complex. RUBISCO spacer regions are even more highly conserved among closely related colonial Phaeocystis species and are identical in Phaeocystis antarctica, Phaeocystis pouchetii and two warm-temperate strains of Phaeocystis globosa, with a single base substitution in two cold-temperate strains of P. globosa. The RUBISCO spacer sequences from two predominantly unicellular Phaeocystis isolates from the Mediterranean Sea and PLY 559 were clearly different from other Phaeocystis strains. In contrast, ITS1 exhibited substantial inter- and intraspecific sequence divergence and showed more resolution among the taxa. Distinctly different copies of the ITS1 region were found in P. globosa, even among cloned DNA from a single strain, suggesting that it is a species complex and making this region unsuitable for phylogenetic analysis in this species. However, among nine P. antarctica strains, four ITS1 haplotypes could be separated. Using the branching order in the ITS1 tree we have attempted to trace the biogeographic history of the dispersal of strains in Antarctic coastal waters.     

A mesocosm study of Phaeocystis globosa (Prymnesiophyceae) population dynamics: II. Significance for the microbial community

The impact of Phaeocystis globosa population decline on the microbial community was studied during a mesocosm experiment, with irradiance regime and inorganic N:P ratios (4, 16, and 44) as controlling factors. Heterotrophic bacterial activity was closely related to enhanced (viral) lysis rates of P. globosa cells and disintegration of the colonies. Up to 85% of the bacterial C demand could be supplied by P. globosa-specific cellular C release. The bacterial populations with high DNA content became dominant (>70% of total). The bacterial community showed a rapid shift in composition to take advantage of the changing conditions during the demise of the P. globosa bloom. Members of the Alphaproteobacteria and the Bacteroidetes group emerged directly upon bloom decay. Multidimensional scaling analysis in conjunction with DGGE fingerprinting implied that clustering was more related to the availability of organic carbon (the collapse of the P. gobosa bloom) than to the nature of the phytoplankton growth-controlling nutrient. Reduced irradiance delayed the development of the P. globosa population and subsequently changes in the bacterial community composition. Disintegration of P. globosa colonies resulted in the formation of transparent exopolymeric particles (TEP) and aggregates, more so under P-depletion than under N-deficient conditions. The colonial matrix transformed into big aggregates under P-depleted conditions but remained largely as ghost colonies under N-depleted conditions. In the mesocosm with initial nitrogen and phosphorus supplied in the Redfield ratio, features intermediate to conditions with either N- or P-depletion were observed. It was hypothesized that TEP affected microbial population dynamics directly through bacterial colonization and indirectly through scavenging of predators and viruses.     

A mesocosm study of Phaeocystis globosa population dynamics: I. Regulatory role of viruses in bloom control

The regulatory role of viruses on population dynamics of the prymnesiophyte Phaeocystis globosa was studied during a mesocosm experiment in relation to growth and loss by microzooplankton grazing and cell lysis. The mesocosms were conducted under varying light conditions (20 and 150 μmol photons m⁻² s⁻¹) and nutrient regime (inorganic nitrogen to phosphorus ratios of 4, 16 and 44). Overall, viruses infecting P. globosa (PgV) were found to be an important cause of cell lysis (30–100% of total lysis) and a significant loss factor (7–67% of total loss). We demonstrate that the morphology of P. globosa cells (solitary versus colonial) differently regulated viral control of P. globosa bloom formation. Reduced irradiance (20 μmol photons m⁻² s⁻¹) was provided for 11 days to select for the solitary cell morphotype. Viruses were able to restrict P. globosa bloom formation even after irradiance became saturating again (150 μmol photons m⁻² s⁻¹). Saturating light conditions from the start of the experiment allowed colony formation and because the colony-morphotype acted as a mechanism reducing viral infection bloom formation succeeded. Nutrient depletion, however, affected specifically the colonies that disintegrated while releasing single cells. Virus infection of these solitary cells resulted in the termination of the bloom. The nature of phytoplankton growth-limiting nutrient (nitrate and/or orthophosphate) did not seem to noticeably affect the level of viral control.     

Inhibition of copepod feeding by exudates and transparent exopolymer particles (TEP) derived from a Phaeocystis globosa dominated phytoplankton community

We investigated if (1) dissolved compounds excreted by Phaeocystis globosa and (2) transparent exopolymer particles (TEP) formed from carbohydrates excreted into the water affect the feeding of nauplii and females of the calanoid copepod Temora longicornis during a P. globosa bloom. Copepod grazing on the diatom Thalassiosira weissflogii in the presence of these possible grazing deterrents was measured during three successive weeks of a mesocosm study, simulating the development of a P. globosa bloom. Our results demonstrate no indication for the presence of feeding deterrents in the dissolved phase, but a strong inhibitory effect of transparent exopolymer particles (TEP) on the consumption of algae by both nauplii and adult copepods. The inhibitory effect of TEP was connected to the accumulation of DOM during the progress of the bloom. We suggest that a reduction in the grazing pressure of zooplankton may increase the survival of the liberated single cells during disruption of colonies and allow seeding populations to persist. Furthermore, P. globosa reduces the trophic efficiency of the food web not only by withdrawal of its colonies from grazing but also by a relaxation of the grazing pressure on co-occurring phytoplankton and by alteration of the food web structure via TEP production.     

The colonization of two Phaeocystis species (Prymnesiophyceae) by pennate diatoms and other protists: a significant contribution to colony biomass

The association of Phaeocystis spp. with small pennate diatoms during three Phaeocystis-dominated spring blooms were investigated in the Eastern English Channel (2003 and 2004) and in coastal waters of Western Norway during a mesocosm experiment (2005). In each of these studies, colonization of the surface of large Phaeocystis spp. colonies by small needle-shaped diatoms (Pseudo-nitzschia spp.) were observed. In the English Channel the diatom Pseudo-nitzschia delicatissima colonized the surface of large (>100 μm) Phaeocystis globosa colonies. The abundance of Pseudo-nitzschia delicatissima reached 130 cells per colony and formed up to 70% of the total carbon associated with Phaeocystis cells during late bloom stages. In Norwegian waters, the surface of large (>250 μm) Phaeocystis pouchetii colonies were colonized by Pseudo-nitzschia cf. granii var. curvata and to a lesser degree by other phytoplankton and protist species, although the abundance of these diatoms was never greater than 40 cells per colony. Based on these observations we suggest that diatoms utilize Phaeocystis colonies not only as habitat, but that they are able to utilize the colonial matrix as a growth substrate. Furthermore, these observations indicate that a considerable fraction of biomass (chlorophyll) associated with Phaeocystis colonies, especially large colonies concerned with intense and prolonged blooms, are due to co-occurring plankton species and not exclusively Phaeocystis cells.     

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。     

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.     

Vernal sedimentation trends in north Norwegian fjords: temporary anomaly in 234Th particulate fluxes related to Phaeocystis pouchetii proliferation

We report data of a naturally occurring radionuclide, ²³⁴Th, an in situ tracer, to investigate vertical export of biogenic matter during a vernal bloom of Phaeocystis pouchetii in the fjords of northern Norway. To optimise sampling of different stages of the bloom, three fjords with increasing oceanic influence (Balsfjord, Malangen fjord and Ullsfjord, respectively) were investigated in April 1997. Contrasting situations were encountered between the three fjords: the proliferation of P. pouchetii in Ullsfjord surface waters coincided with a drastic reduction of particulate ²³⁴Th fluxes in traps, although particulate organic carbon (POC) and dimethylsulphoniopropionate (DMSP) were exported and ²³⁴Th was available in surface waters. When large colonies make up a significant fraction of the vertical flux, as observed in Ullsfjord in April 1997, there may be a large and rapid change in the POC/²³⁴Th ratio, further complicating the use of ²³⁴Th as a tracer for POC export. The results suggest that the proliferation of Phaeocystis pouchetii during vernal bloom could temporary increase OC/²³⁴Th ratio of particles and delay the particulate export of ²³⁴Th, and probably of other particle-reactive species, from surface waters.     

Haemolytic activity of live Phaeocystis pouchetii during mesocosm blooms

Chemical defence is a potential mechanism contributing to the success of Phaeocystis species that repeatedly dominate the phytoplankton in coastal areas. Species within the genus Phaeocystis have long been suspected of imposing negative effects on co-occurring organisms. Recently a number of toxins have been extracted and identified from Phaeocystis samples, but it is not clear if they do enhance the competitive advantage of Phaeocystis species. In the present study the cytotoxic impact of live Phaeocystis pouchetii to human blood cells in close proximity, regardless of the nature of the responsible mechanism, was quantified using a bioassay. Haemolytic activity was measured during blooms of P. pouchetii in mesocosms. These environments were chosen to mimic natural conditions including chemically mediated interactions that could trigger defensive and/or allelopathic responses of Phaeocystis. Haemolytic activity correlated with P. pouchetii numbers and was absent during the preceding diatom bloom. Samples containing live P. pouchetii cells showed the highest activity, while filtered sea water and cell extracts were less haemolytic or without effect. Dose-response curves were linear up to 70% lysis, and haemolysis in samples containing live P. pouchetii cells reached EC₅₀ values comparable to known toxic prymnesiophytes (1.9 * 10⁷ cells l⁻¹). Haemolytic activity was enhanced by increased temperature and light. The results indicate that unprotected and thus presumably vulnerable cells present in a P. pouchetii bloom may lyse within days.     

Long-term development of the crustacean plankton in the Saidenbach Reservoir (Germany) – changes,causes, consequences

The rates of development and food intake of the copepod Temora longicornis (Müller) were studied using artificial blooms of Phaeocystis globosa Scherffel under different conditions of nutrient limitation. Mesocosms with 800 l of natural seawater were manipulated by inoculation with cultured P. globosa and by addition of nitrogen and/or phosphorus, to obtain N- or P-limited blooms of P. globosa. During development and ageing of these blooms, water from the mesocosms was used as medium for incubation of nauplii of T. longicornis. Only moderate rates of naupliar development as well as high rates of mortality were observed, irrespective of major differences of nutrient conditions and density of P. globosa. Grazing by the nauplii on P. globosa seemed to be low, suggesting a low food quality of this alga at all physiological conditions studied. The results of this study indicate a low capability of T. longicornis nauplii for control of nuisance algal blooms caused by P. globosa.     

Zooplankton grazing on Phaeocystis: a quantitative review and future challenges

The worldwide colony-forming haptophyte phytoplankton Phaeocystis spp. are key organisms in trophic and biogeochemical processes in the ocean. Many organisms from protists to fish ingest cells and/or colonies of Phaeocystis. Reports on specific mortality of Phaeocystis in natural plankton or mixed prey due to grazing by zooplankton, especially protozooplankton, are still limited. Reported feeding rates vary widely for both crustaceans and protists feeding on even the same Phaeocystis types and sizes. Quantitative analysis of available data showed that: (1) laboratory-derived crustacean grazing rates on monocultures of Phaeocystis may have been overestimated compared to feeding in natural plankton communities, and should be treated with caution; (2) formation of colonies by P. globosa appeared to reduce predation by small copepods (e.g., Acartia, Pseudocalanus, Temora and Centropages), whereas large copepods (e.g., Calanus spp.) were able to feed on colonies of Phaeocystis pouchetii; (3) physiological differences between different growth states, species, strains, cell types, and laboratory culture versus natural assemblages may explain most of the variations in reported feeding rates; (4) chemical signaling between predator and prey may be a major factor controlling grazing on Phaeocystis; (5) it is unclear to what extent different zooplankton, especially protozooplankton, feed on the different life forms of Phaeocystis in situ. To better understand the mechanisms controlling zooplankton grazing in situ, future studies should aim at quantifying specific feeding rates on different Phaeocystis species, strains, cell types, prey sizes and growth states, and account for chemical signaling between the predator and prey. Recently developed molecular tools are promising approaches to achieve this goal in the future.