Two new subfamilies of DNA mismatch repair proteins (MutS) specifically abundant in the marine environment

MutS proteins are ubiquitous in cellular organisms and have important roles in DNA mismatch repair or recombination. In the virus world, the amoeba-infecting Mimivirus, as well as the recently sequenced Cafeteria roenbergensis virus are known to encode a MutS related to the homologs found in octocorals and ɛ-proteobacteria. To explore the presence of MutS proteins in other viral genomes, we performed a genomic survey of four giant viruses (‘giruses'') (Pyramimonas orientalis virus (PoV), Phaeocystis pouchetii virus (PpV), Chrysochromulina ericina virus (CeV) and Heterocapsa circularisquama DNA virus (HcDNAV)) that infect unicellular marine algae. Our analysis revealed the presence of a close homolog of Mimivirus MutS in all the analyzed giruses. These viral homologs possess a specific domain structure, including a C-terminal HNH-endonuclease domain, defining the new MutS7 subfamily. We confirmed the presence of conserved mismatch recognition residues in all members of the MutS7 subfamily, suggesting their role in DNA mismatch repair rather than DNA recombination. PoV and PpV were found to contain an additional type of MutS, which we propose to call MutS8. The MutS8 proteins in PoV and PpV were found to be closely related to homologs from ‘Candidatus Amoebophilus asiaticus'', an obligate intracellular amoeba-symbiont belonging to the Bacteroidetes. Furthermore, our analysis revealed that MutS7 and MutS8 are abundant in marine microbial metagenomes and that a vast majority of these environmental sequences are likely of girus origin. Giruses thus seem to represent a major source of the underexplored diversity of the MutS family in the microbial world.     

Growth characteristics of flagellated cells of Phaeocystis pouchetii revealed by diel changes in cellular DNA content

The present study reports on effects of different light:dark periods, light intensities, N:P ratios and temperature on the specific growth rate of flagellated cells of Phaeocystis pouchetii in culture. The specific growth rate was estimated by diel changes in cellular DNA content. The cellular DNA content and cell cycle of flagellated cells of P. pouchetii are shown, and the importance of light:dark period in cell division is demonstrated. Diel patterns of the cellular DNA content showed that cell division was confined to the dark period. The cells dealt with more than one division per day by rapid divisions shortly after each other.The specific growth rates (μ_DNA) based on the DNA cell cycle model were in close agreement with specific growth rates (μ_Cell) determined from cell counts. The temperature affected the specific growth rates (multiple regression, p < 0.01) and were higher at 5 °C (μ ≤ 2.2 d⁻¹) than at 10 °C (μ ≤1.6 d⁻¹). Increasing the light:dark period from 12:12 h to 20:4 h affected the specific growth rate of P. pouchetii at the lower temperature tested (5 °C) (multiple regression, p < 0.01), resulting in higher specific growth rates than at 10 °C. At 10 °C, the effect of light:dark period was severely reduced. Neither light nor nutrients could compensate the reduction in specific growth rates caused by elevated temperature. The specific growth rates was not affected by the N:P ratios tested (multiple regression, p = 0.21). The experiments strongly suggest that the flagellated cells have a great growth potential and could play a dominating role in northern areas at increased day length.     

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.     

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.     

Effect of some environmental factors on cyanophage AS-1 development in Anacystis nidulans

The development cycle of the cyanophage AS-1 was studied in the host blue-green alga, Anacystis nidulans, under conditions that impair photosynthesis and under various light/dark regimes. Under standard conditions of incubation the 16-h development cycle consisted of a 5-h eclipse period and an 8-h latent period. Burst size was decreased by dark incubation to 2% of that observed in the light. An inhibitor of photosystem II, 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU), reduced the burst size to 27% of that of the uninhibited control, whereas cyanophage production was completely abolished by carbonyl-cyanide m-chlorophenyl hydrazone (CCCP), an inhibitor of photosynthetic electron transport. Dark incubation of infected cells decreased the latent period by 1–2 h and the eclipse period by 1 h, once the cultures were illuminated. This suggests that adsorption took place in the dark. Intracellular growth curves indicated that light is necessary for viral development. Infected cells must be illuminated at least 13 h to produce a complete burst at the same rate as the continuously illuminated control. Low light intensities retarded the development cycle, and at lowest light intensities no phage yield was obtained. AS-1 is highly dependent on host cell photophosphorylation for its development.List of Abbreviations CCCP Carbonyl-cyanide m-chlorophenyl hydrazone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl urea - m.o.i. multiplicity of infection - O.D. optical density - PFU plaque-forming unit Dedicated to Prof. Roger Y. Stanier on the occasion of his 60th birthday     

Oscillatory behavior of control-systems of calcium homeostasis in chickens

Computer simulation of calcium homeostasis in chicks predicted an oscillatory behavior of bone calcium flow and kidney 25-hydroxyvitamin D₃-1 hydroxylase with a periodicity of 56 h and a 9 h phase difference between the two signals. In growing chickens subjected to a light: dark cycle of 22:2 h, and intravenously dosed with ⁴⁵Ca, the temporal changes in plasma ⁴⁵Ca could be described by an exponential decline with superimposed diurnal oscillations. The activity of the renal 25-hydpoxyvitamil D₃-1-hydroxylase in chicks subjected to a 12:12 h light: dark cycle ALSO followed diupnal oscillations, with a ladir at the beginning of the light period and a peak 12 h later. The production of 1,25-dihydroxyvitamin D₃ by primary cultures of chicken kidney cells mscillated with a periodicity of 5.6 h or shorter. It is suggested that despite the differences in phase and periodicity between the simulation predictions and actual results, the oscillations in both 1-hydroxylase and bone calcium flow could be coupled through the hormonal systems involved in regulation of plasma calcium.     

Differential patterns of mitochondrial,chloroplastic and nuclear DNA synthesis in the synchronous cell cycle of Chlamydomonas reinhardtii

Nuclear DNA (ncDNA) synthesis in Chlamydomonas reinhardtii was measured by both ³²P[or-thophosphoric acid] (³²P) and [¹⁴C]adenine incorporation and found to be highly synchronous. Ca. 85% of incorporation was confined to the first 6 h of the dark period of a synchronized regime consisting of an alternating light-dark period of 12 h each. In contrast, no such synchronous incorporation pattern was found for chloroplast (cp) and mitochondrial (mt) DNAs in the same cell population. These two organellar DNAs also exhibited different ³²P-incorporation patterns in the cell cycle. Considerable amounts of ³²P were incorporated into cpDNA throughout the light-dark synchronous cycle under both mixo- and phototrophic growth conditions, although the second 6-h light period under phototrophy showed an increase not apparent under mixotrophy. This change in growth conditions did not affect ³²P incorporation into mtDNA, which was found throughout the cell cycle, with a modest peak in the first 6-h of the dark period. The pattern of [³H]thymidine incorporation into cpDNA was also determined. Under synchronous phototrophic conditions, this pattern was quite different from that obtained with ³²P. Most [³H]thymidine incorporation occurred during the light period of the synchronous cycle; this period had been shown previously by density transfer experiments to be the time of cpDNA duplication. Such preferential [³H]thymidine incorporation into cpDNA in the light period was not observed under mixotrophic synchronous growth conditions; in these, [³H]thymidine incorporation was detected throughout the cell cycle. This lack of coincidence between the patterns of ³²P- and of [³H]thymidine incorporation into cpDNA during the synchronous cell cycle indicates that in addition to replication, the considerably reiterated organelle-DNA molecules may also regularly undergo an extensive repair process during each cell cycle.     

Nitrogenase activity in the non-heterocystous cyanobacterium Oscillatoria sp. grown under alternating light-dark cycles

The non-heterocystous cyanobacterium Oscillatoria sp. strain 23 fixes nitrogen under aerobic conditions. If nitrate-grown cultures were transferred to a medium free of combined nitrogen, nitrogenase was induced within about 1 day. The acetylene reduction showed a diurnal variation under conditions of continuous light. Maximum rates of acetylene reduction steadily increased during 8 successive days. When grown under alternating light-dark cycles, Oscillatoria sp. fixes nitrogen preferably in the dark period. For dark periods longer than 8 h, nitrogenase activity is only present during the dark period. For dark periods of 8 h and less, however, nitrogenase activity appears before the beginning of the dark period. This is most pronounced in cultures grown in a 20 h light – 4 h dark cycle. In that case, nitrogenase activity appears 3–4 h before the beginning of the dark period. According to the light-dark regime applied, nitrogenase activity was observed during 8–11 h. Oscillatoria sp. grown under 16 h light and 8 h dark cycle, also induced nitrogenase at the usual point of time, when suddenly transferred to conditions of continuous light. The activity appeared exactly at the point of time where the dark period used to begin. No nitrogenase activity was observed when chloramphenicol was added to the cultures 3 h before the onset of the dark period. This observation indicated that for each cycle, de novo nitrogenase synthesis is necessary.     

The contribution of single and colonial cells of Phaeocystis pouchetii to spring and summer blooms in the north-eastern North Atlantic

Studies of the phytoplankton ecology in different localities in north-Norwegian fjords, the White Sea and the Barents Sea were carried out in spring and early summer to investigate the contribution of single and colonial stages of Phaeocystis pouchetii to phytoplankton abundance. Three different types of flagellated and four colonial cells were observed in all localities. P. pouchetii was rare under the ice of the Barents and White Seas, but their abundance increased rapidly during ice retreat. Single cell C dominated over colonial cell C, often by 50 times or more. The highest share of colonial cells was encountered in April in northern Norwegian fjords, in May in the Barents Sea and in May–June in the White Sea. At times the single cell dominated the total P. pouchetii biomass in Balsfjord (April 1999, 2001) with hardly any colonies present. In the White Sea colonies of P. pouchetii were less abundant than in the other regions. Cell carbon of P. pouchetii colonies appears never to be as dominating in the north-eastern North Atlantic as P. globosa blooms in coastal regions such as the southern North Sea. However, the lobal matrix of P. pouchetii colonies appears to be less solid than that of P. globosa and partly dissolution of the colony matrix during handling and storage of fixes samples induces uncertainty about the absolute numbers of P. pouchetii colonial cell counts. Despite of that, single cells of P. pouchetii seem to dominate significantly over colonial cell biomass at most sites and during some years and in some regions colonial cells seem rare. We speculate that top-down regulation of Phaeocystis spp. blooms possibly determines the ratio between single and colonial cells.     

Stage-dependent localization of LHCP II apoprotein in the Golgi of synchronized cells of Euglena gracilis by immunogold electron microscopy

We have localized LHCP II apoprotein in the Golgi and thylakoids of Euglena gracilis Klebs var. bacillaris Cori and strain Z Pringsheim by electron microscopy using a specific antibody and protein A-gold. Using synchronized cells (light, 14 h:dark, 10 h) we show that thylakoids are always immunoreactive. There is no reaction in the Golgi at 0 h (the beginning of the light period) but immunoreaction appears in the Golgi soon thereafter, rises to a peak at 8 h and declines to zero by 16 h (2 h into the dark period). The peak in immunoreaction in the Golgi immediately precedes the peak in cellular ¹⁴C-labeling of thylakoid LHCP II apoprotein seen by Brandt and von Kessel (Plant Physiol. (1983) 72, 616), supporting our suggestion that processing in the Golgi precedes deposition of LHCP apoprotein in the thylakoids. Substitution of preimmune serum for antiserum eliminates the immunoreaction in the Golgi, and thylakoids of synchronized cells of mutant Gr₁BSL which lacks LHCP II apoprotein show no immunoreaction in the Golgi or thylakoids at any stage. Random observations indicate that the compartmentalized osmiophilic structure (COS) shows an immunoreaction with anti-LHCP II apoprotein antibody at 1 h into the light period (when the Golgi is not immunoreactive) and at 10 h into the light period (when the Golgi is fully reactive), suggesting that the COS remains immunoreactive throughout the cell cycle.     

PHOSPHATE UPTAKE KINETICS IN EUGLENA GRACILIS (Z) (EUGLENOPHYCEAE) GROWN ON LIGHT/DARK CYCLES.' I. SYNCHRONIZED BATCH CULTURES1

The characteristics of phosphate uptake in synchronized populations of Euglena gracilis Klebs (Z) were studied. The cells were grown autotrophically in batch culture and synchronized with a cycle of 14:10 LD. Incorporation of P was nonlinear with time for the first 2 h of incubation over a wide range of P concentrations and completely inhibited by darkness. The kinetics of P uptake as a function of P concentration were triphasic between 0 and 100 μM PO₄, obeying Michaelis-Menten kinetics over the 0–3 μM PO₄ range-only. Uptake velocity increased linearly with, concentration above 3 μM PO₄. The kinetics of P uptake varied with stage in the cell cycle. The half-saturation constant for uptake at the lower concentrations oscillated between 0.7 and 2.8 μM PO₄, reaching a peak immediately before the onset of cell division (beginning of the dark period). V_max was largest in the middle of the light period, as was the slope of the linear portion of the kinetic pattern. Further analysis of the kinetics suggests that changes in this slope are responsible for the oscillation in K_s values calculated for the lower concentrations. This analysis assumes 2 uptake mechanisms, one which saturates at low concentrations of phosphate, and one which is nonsaturable over the entire concentration range examined.     

Diurnal differences in the supply of glucomannans and xylans to innermost surface of cell walls at various developmental stages from cambium to mature xylem in Cryptomeria japonica

Summary. We investigated the diurnal differences in the innermost surface of tracheid cell walls at various developmental stages from cambium to mature xylem. Cryptomeria japonica saplings were cultivated in a growth chamber with a light cycle set at 14 h of light and 10 h of darkness. Samples were collected from the saplings during both the light and dark periods. The innermost surface of cell walls was immunogold-labeled with anti-glucomannan or anti-xylan antiserum and was observed by field emission scanning electron microscopy. Diurnal differences in the aspect of the innermost surface of cell walls were seen only in S₂-layer-forming tracheids; cellulose microfibrils were clearly evident during the light period, and amorphous material containing glucomannans and xylans was prevalent during the dark period. Cellulose microfibrils were present at the primary-wall formation and S₁-layer-forming stages, and many warts were observed in the mature tracheids, regardless of the time of sampling. The densities of labeled glucomannans on the innermost surface of cell walls in S₁- and S₂-forming tracheids and of labeled xylans in S₂-forming tracheids during the dark period were significantly higher than those during the light period. These results suggest a diurnal periodicity in the supply of cell wall matrix containing hemicellulose to the innermost surface of developing secondary walls. Correspondence and reprints: Laboratory of Bio-material Physics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan. Present address: Chair of Climate Change Science for Forestry and Water Resources, Graduate School of Science and Technology, Niigata University, Niigata, Japan.     

Rhabdom degradation in white-eyed and wild-type crayfish after long term dark adaptation

Summary Rhabdom degradation has been compared in groups of wild-type (WT) and white-eyed mutant (WEM) crayfish exposed to a 1212 LD cycle after 1 month of continuous darkness. Light and electron microscopic quantitative analysis of the lysosome-related bodies (LRB) formed during rhabdom degradation show that WEM photore-ceptors produced few LRB during the first 12 h of light exposure, but the microvilli in the rhabdom became severely disrupted. An increase in LRB production began in the WEM after the initial light-dark transition, and continued during the first dark period, as well as throughout the second light cycle. Analysis of fluctuations in specific LRB types shows that multivesicular bodies (MVB) were the major contributor to the total organelle population seen in the WEM, and lamellar bodies (LB) showed a significant drop following light onset of the second light cycle.Wild-type crayfish maintained in long-term dark before exposure to cyclic lighting showed a characteristic increase in LRB production in the light and a decline in the dark. However, LRB persisted throughout the light period and did not show a characteristic decline during the second 6 h in the light. This exaggerated degradative response in the WT animals was repeated during the second light cycle.Abbreviations WEM white-eyed mutant - LRB lysosome-related bodies - MVB multivesicular bodies - LB lamellar bodies - CB combination bodies     

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.     

The secretory dynamics of the CHH-producing cell group in the eyestalk of the crayfish,Astacus leptodactylus,in the course of the day/night cycle

Summary The secretory dynamics of the Crustacean Hyperglycemic Hormone (CHH)-producing cells in the eyestalk of the crayfish Astacus leptodactylus were studied during the daily cycle (12 h light/12 h dark). The different secretory stages of individual cells were determined by means of immunocytochemistry combined with morphometric analysis at the light-microscopic level. The data obtained were correlated with the 24-h rhythmicity of blood glucose concentration. The results suggest the following hypothesis. The synthetic activity of the CHH cells receives a stimulus 2 h before the beginning of the dark period, resulting in a pronounced transfer of CHH granules into the axons. These CHH granules reach the axon terminals after the onset of the dark period. At that time a burst of exocytotic activity occurs, causing a strong release of CHH into the hemolymph. Four hours later this CHH release results in hyperglycemia. The same process, though with less intensity, is repeated and causes a second smaller glucose peak at the beginning of the light period.     

The "Point of no Return" Concept in Cell Division. The Effects of Some Metabolic Inhibitors on Synchronized Chlorella pyrenoidosa

The coarse of growth and cell division in synchronized cultures of Chlorella pyrenoidosa was studied after the addition of metabolic inhibitors at differing times during the cell cycle (14 h light - 10 h darkness with nitrate as nitrogen source. 12 h light: 12 h darkness with urea as nitrogen source). Dinitrophenol (DNP) added to a final concentration of 0.3 mM at any time in the synchronization cycle, the compound remaining in the suspension from the time of addition to the end of the dark period, inhibited spore formation completely. Growth measured as increase in cell volume was less sensitive to the action of the inhibitor. Chloramphenicol (CAP) added dining the 0–5 h interval to a final concentration of 0.1 mM resulted in 80 per cent inhibition of cell division. Similar treatment started at successive times thereafter resulted in a gradual decrease of the inhibition. Treatment at the 14th hour and during the dark period did not affect the sporulation. Similar experiments with 0.9 mM puromycin added at various times during the illumination period gave almost complete inhibition of cell division, while the growth was reduced by only 25 per cent. para-Fluorophenylalanine (p-FPhe) at 3.3 × 10^?2 mM stopped cell division nearly completely irrespective of addition time in the light period. Addition during the dark period also prevented an increase in the number of tree cells. In this case about half of the cells produced spores which were not released. It is concluded that DNP inhibits all stages of preparation for cell division, as well as the division process itself. With CAP a genuine transition point of preparation for cell division was observed, although its interpretation as related to protein synthesis is somewhat uncertain. With puromycin and p-FPhe no transitions were observed.     

LIGHT/DARK-PHASED CELL DIVISION IN EUGLENA GRACILIS (Z) (EUGLENOPHYCEAE) IN PO4-LIMITED CONTINUOUS CULTURE1

Eugene gracilis Klebs (Z) was grown in a cyclostat (continuous culture on a light/dark cycle) at growth limiting levels of phosphate. Cell division was restricted to the dark period regardless of the proportion of the cells dividing during each 24 h period. Growth rate, as reflected by the amplitude of the cell density oscillation, was correlated with dilution rate. The width of the division gate was analyzed using a phasing index and found to be narrowest at dilution rates where the mean generation time of the cell population was an even multiple of 24 h. The effect was attributed to enhanced phasing of the cell division process by the biological clock of Euglena. Residual phosphate levels in the cyclostat were less than 0.3 μM PO₄ at all submaximal growth rates. Cellular phosphorus concentration increased with dilution rate as described by a hyperbola saturating at D_max= 0.74 day⁻¹ with 8 × 10⁻⁸μM P/cell as the minimum intracellular phosphorus concentration for growth. The results are discussed, in terms of the inherent similarities and differences between a cyclostat and a steady state chemostat, and the advantages of the cyclostat for studies in phytoplankton ecology.     

SIMILARITIES IN THE ASEXUAL REPRODUCTION OF THE OCEANIC DINOFLAGELLATES, PYROCYSTIS FUSIFORMIS, PYROCYSTIS LUNULA, AND PYROCYSTIS NOCTILUCA1

Three cultured species of Pyrocystis (Dinoccoccales) reproduced asexually by forming 2 (or 1) aplanospores or zoospores inside the parent cell wall. In all 3 species these small reproductive cells, although they may not resemble the parent cells, swell up rapidly (～ 10 min) to the approximate size and shape of the parent cell. These swollen cells become new vegetative cells. The above asexual process is the only way by which cells numbers increase in our cultures. Pyrocystis lunula was propagated at the lunula stage of the life cycle. The nonmotile crescent-shaped cells produced reproductive cells that were Gymnodinium-shaped and had, in some cases, a trailing flagellum. With P. fusiformis and P. noctiluca, the reproductive cells were not flagellated. With P. fusiformis, these bodies had a pronounced equatorial constriction like a girdle, while in P. noctiluca the “girdle” was an inconspicuous feature if present. With P. noctiluca and P. fusiformis on a 12:12 ld cycle, reproductive cells were formed early in the dark period and they swelled up at the beginning of the light period. Reproduction of P. lunula was not well phased in our experiments, with reproductive cells developing at the end of the light period and the end of the dark period.     

DIEL CHANGES IN THE COMPOSITION OF PHOTOSYNTHETIC PIGMENTS AND CELLULAR CARBON AND NITROGEN IN PYRAMIMONAS PARKEAE (PRASINOPHYCEAE)1

Diel changes in mean cell volume, cellular carbon (carbon content per cell), cellular Chl a, C/N ratio, Chl a/carbon ratio and pigment composition were determined for an axenic clonal culture of Pyramimonas parkeae Norris et Pearson through three 12:12 h LD cycles in a laboratory culture tank of 1 m³. Mean cell volume and cellular C, N and most pigments increased during the light period as a result of photosynthesis and decreased with an increase in cell density by phased cell division during the dark period. Chi a and Chi b increased in a parallel manner during the light period. Increases in the diel synthesis pattern of carotenoids varied. Violaxanthin and lutein content increased for a few hours at the beginning of the light period and preceeded that of neoxanthin. The diel synthesis pattern of neoxanthin was similar to that of Chi a. Increases of loroxanthin and its ester form were slower than that of Chi a at the beginning of the light period. A net increase of α-carotene was observed during the dark period. Mass spectroscopy of carotenoid structure showed a new xanthophyll, loroxanthin dodecenoate, in this species.     

Influence of medium frequency light/dark cycles of equal duration on the photosynthesis and respiration of Chlorella pyrenoidosa

Chlorella pyrenoidosa was grown in three continuous cultures each receiving a different light regime during the light period of a diurnal cycle. Hourly samples taken during the light period were subjected to medium frequency light/dark oscillations of equal duration, ranging from 3 to 240 seconds. The oxygen consumption and production of each sample were measured with an oxygen electrode in a small oxygen chamber. Although the light/dark cycles had little overall influence on photosynthetic activity, the microalgae appeared to adapt to the light regime to which they were subjected. Large differences were found between the maximum chlorophyll-specific production rates (P _infmax ^supB ), the chlorophyll-specific production rates (P^B) and the respiration rates between the cultures and treated subsamples. Respiration rates increased during the light period, whilst P^B either increased, or had a mid light period minimum or maximum. The culture which received an hourly light oscillation during the light period had the highest P _infmax ^supB and lowest respiration rates, and it is suggested that these algae react as in nature, whereas either a sinusoidal or a block light pattern is unnatural. The latter light regime is commonly used in laboratory studies.