Isolation of membrane bound light-harvesting-complexes from the dinoflagellates Heterocapsa pygmaea and Prorocentrum minimum.

We have isolated Chl a-Chl c-carotenoid binding proteins from the dinoflagellates Prorocentrum minimum and Heterocapsa pygmaea grown under high (500 μmol m(-2) s(-1), HL) and low (35 μmol m(-2) s(-1), LL) light conditions. We compared various isolation procedures of membrane bound light harvesting complexes (LHCs) and assayed the functionality of the solubilized proteins by determining the energy transfer efficiency from the accessory pigments to Chl a by means of fluorescence excitation spectra. The identity of the newly isolated protein-complexes were confirmed by immunological cross-reactions with antibodies raised against the previously described membrane bound Chl a-c proteins (Boczar et al.

High abundance of Archaea in Antarctic marine picoplankton.

Archaea (archaebacteria) constitute one of the three major evolutionary lineages of life on Earth. Previously these prokaryotes were thought to predominate in only a few unusual and disparate niches, characterized by hypersaline, extremely hot, or strictly anoxic conditions. Recently, novel (uncultivated) phylotypes of Archaea have been detected in coastal and subsurface marine waters, but their abundance, distribution, physiology and ecology remain largely undescribed.

Calibration of an integrating sphere for determining the absorption coefficient of scattering suspensions.

Measuring the absolute absorption of suspensions of absorbing particles with unknown scattering characteristics is not possible in conventional spectrophotometers or in integrating spheres that have the sample located outside the sphere. A method for the calibration and use of an integrating sphere with a centrally located sample to measure absolute absorption coefficients of scattering suspensions is presented. Under the tested conditions the integrating sphere used in this study was insensitive to changes in the scattering coefficient of the sample but had a nonlinear response to increasing absorption of the sample, which could be corrected with an empirically derived function.

Characterization of two full-length cDNA sequences encoding for apoproteins of peridinin-chlorophyll a-protein (PCP) complexes.

Characterizations are presented for RNA, 2 cDNA libraries, and 2 full-length cDNA sequences encoding for photosynthetic light-harvesting peridinin-chlorophyll a-protein (PCP) in the dinoflagellate Heterocapsa pygmaea. Subsequent analyses of the PCP system also indicate that (1) it is represented by multiple nuclear encoded genes, (2) a subset of mRNAs encoding for PCP apoproteins are regulated by growth irradiance, (3) PCP preproteins are larger than the mature apoproteins, and (4) PCP cDNA clones sequenced thus far contain a conserved region but are not identical. Results are discussed in the context of photoadaptation in dinoflagellates.

Ozone depletion: ultraviolet radiation and phytoplankton biology in antarctic waters.

The springtime stratospheric ozone (O3) layer over the Antarctic is thinning by as much as 50 percent, resulting in increased midultraviolet (UVB) radiation reaching the surface of the Southern Ocean. There is concern that phytoplankton communities confined to near-surface waters of the marginal ice zone will be harmed by increased UVB irradiance penetrating the ocean surface, thereby altering the dynamics of Antarctic marine ecosystems. Results from a 6-week cruise (Icecolors) in the marginal ice zone of the Bellingshausen Sea in austral spring of 1990 indicated that as the O3 layer thinned: (i) sea surface- and depth-dependent ratios of UVB irradiance (280 to 320 nanometers) to total irradiance (280 to 700 nanometers) increased and (ii) UVB inhibition of photosynthesis increased.

Light Regulation of Peridinin-Chlorophyll a-Protein (PCP) Complexes in the Dinoflagellate, Glenodinium sp. : Use of Anti-Pcp Antibodies to Detect Pcp Gene Products in Cells Grown in Different Light Conditions.

As a step toward developing the tools needed to study the molecular bases of light regulation of gene expression in dinoflagellates, light-harvesting peridinin-chlorophyll a-protein (PCP) complexes from Glenodinium sp. were purified and used to generate anti-PCP antibodies. Affinity purified anti-PCP antibodies were isolated from the crude anti-PCP antiserum resulting in improved specificity of immune reactions.

Chlorophyll-Protein Complexes from the Red-Tide Dinoflagellate, Gonyaulax polyedra Stein : Isolation, Characterization, and the Effect of Growth Irradiance on Chlorophyll Distribution.

A comparision of high (330 microeinsteins per meter squared per second) and low (80 microeinsteins per meter squared per second) light grown Gonyaulax polyedra indicated a change in the distribution of chlorophyll a, chlorophyll c(2), and peridinin among detergent-soluble chlorophyll-protein complexes. Thylakoid fractions were prepared by sonication and centrifugation. Chlorophyll-protein complexes were solubilized from the membranes with sodium dodecyl sulfate and resolved by Deriphat electrophoresis.

Changes in Photosystem II Account for the Circadian Rhythm in Photosynthesis in Gonyaulax polyedra.

Cell-free extracts that show activity in photosynthetic electron flow have been prepared from the unicellular dinoflagellate, Gonyaulax polyedra. Electron flow, as O(2) uptake, was measured through both photo-system I and II from water to methyl viologen, through photosystem I alone from reduced 2,6-dichlorophenol indophenol to methyl viologen which does not include the plastoquinone pool or from duroquinol to methyl viologen which includes the plastoquinone pool. Electron flow principally through photosystem II was measured from water to diaminodurene and ferricyanide, as O(2) evolution.

Photosynthetic characteristics and organization of chlorophyll in marine dinoflagellates.

The photosystem I reaction center complex, the P-700-chlorophyll a-protein, has been isolated from the photosynthetic membranes of two marine dinoflagellates, Gonyaulax polyedra and Glenodinium sp., by detergent solubilization with Triton X-100. The complexes isolated from the two species were indistinguishable, exhibiting identical absorption properties (400-700 nm) at both room (300 K) and low (77 K) temperature.

Characterization of Photosynthetic Rhythms in Marine Dinoflagellates: II. Photosynthesis-Irradiance Curves and in Vivo Chlorophyll a Fluorescence.

Using data from light-dark cultures of Gonyaulax polyedra entrained to a 24-hour cycle, whole cell absorption curves and photosynthesis-irradiance curves were constructed for various circadian times. While whole cell absorbance and half-saturation constants of photosynthesis showed no statistical difference that could be directly related to the photosynthetic rhythm, the initial slope of the photosynthesis-irradiance curve was a time-dependent parameter which altered in direct proportion to the change in photosynthetic capacity. The results indicated a temporal change in the relative quantum yield of photosynthesis, and the circadian rhythmicity of light-limited photosynthesis was established under constant conditions.

Characterization of photosynthetic rhythms in marine dinoflagellates: I. Pigmentation, photosynthetic capacity and respiration.

Circadian rhythms in photosynthesis were defined for the first time in the dinoflagellates Glenodinium sp. (M. Bernard strain) and Ceratium furca Ehrenberg (B.

Molecular topology of the photosynthetic light-harvesting pigment complex, peridinin-chlorophyll a-protein, from marine dinoflagellates.

The photosynthetic light-harvesting complex, peridinin-chlorophyll a-protein, was isolated from several marine dinoflagellates including Glenodinium sp. by Sephadex and ion-exchange chromatography. The carotenoid (peridinin)-chlorophyll a ratio in the complex is estimated to be 4:1.

Purification and characterization of peridinin-chlorophyll a-proteins from the marine dinoflagellates Glenodinium sp. and Gonyaulax polyedra.

A peridinin-chlorophyll a-protein complex (PCP) was obtained in large quantity from the marine dinoflagellates, Glenodinium sp. and Gonyaulax polyedra. The chromoproteins have similar molecular weights, 35,500 for Glenodinium sp.

Effects of growth irradiance on the photosynthetic action spectra of the marine dinoflagellate, Glenodinium sp.

Whole cell absorption curves of the marine dinoflagellate Glenodinium sp., cultured at irradiances of 250μW/cm(2) (low light) and 2500μW/cm(2) (high light), were measured and their difference spectrum determined. Absorption by low light grown cells exceeded that of high light grown cells throughout the visible spectrum by a factor which ranged from 2 to 4.

The role of peridinin-chlorophyll a-proteins in the photosynthetic light adaption of the marine dinoflagellate, Glenodinium sp.

The marine dinoflagellate, Glenodinium sp., was cultured at a series of light levels and growth, pigmentation, and photosynthetic rates were compared. Under decreasing light conditions, growth rates decreased, cellular chlorophyll a and peridinin content per cell increased, and maximum cellular photosynthetic rates remained unchanged.