RESPONSE OF PROROCENTRUM MARIAE-LEBOURIAE (DINOPHYCEAE) TO LIGHT OF DIFFERENT SPECTRAL QUALITIES AND IRRADIANCES: GROWTH AND PIGMENTATION1

Growth and pigment concentrations of the, estuarine dinoflagellate, Prorocentrum mariae-lebouriae (Parke and Ballantine) comb. nov., were measured in cultures grown in white, blue, green and red radiation at three different irradiances. White irradiances (400–800 nm) were 13.4, 4.0 and 1.8 W · m^?2 with photon flux densities of 58.7 ± 3.5, 17.4 ± 0.6 and 7.8 ± 0.3 μM quanta · m^?2· s^?1, respectively. All other spectral qualities had the same photon flux densities. Concentrations of chlorophyll a and chlorophyll c were inversely related to irradiance. A decrease of 7- to 8-fold in photon flux density resulted in a 2-fold increase in chlorophyll a and c and a 1.6- to 2.4-fold increase in both peridinin and total carotenoid concentrations. Cells grown in green light contained 22 to 32% more peridinin per cell and exhibited 10 to 16% higher peridinin to chlorophyll a ratios than cells grown in white light. Growth decreased as a function of irradiance in white, green and red light grown cells but was the same at all blue light irradiances. Maximum growth rates occurred at 8 μM quanta · m^?2· s^?1 in blue light, while in red and white light maximum growth rates occurred at considerably higher photon flux densities (24 to 32 μM quanta · m^?2· s^?1). The fastest growth rates occurred in blue and red radiation. White radiation producing maximum growth was only as effective as red and blue light when the photon flux density in either the red or blue portion of the white light spectrum was equivalent to that of a red or of blue light treatment which produced maximum growth rates. These differences in growth and pigmentation indicate that P. mariae-lebouriae responds to the spectral quality under which it is grown.     

Variations in biochemical parameters of Heterocapsa sp. and Olisthodiscus luteus grown in 12:12 light:dark cycles II. Changes in pigment composition

Photosynthetic pigment composition was studied in batch cultures of Heterocapsa sp. and Olisthodiscus luteus growing exponentially in a 12:12 light:dark cycle. Both species divided in the dark. The synthesis of pigments was continuous for both species. However for chlorophyll c and peridinin, in Heterocapsa sp., and chlorophyll c and fucoxanthin, in O. luteus, (pigments belonging to light harvesting complexes) the synthesis was significantly higher during the light period. Concentrations per total cell volume (TCV) of chlorophyll a, chlorophyll c, peridinin and diadinoxanthin in Heterocapsa sp., and chlorophyll a, chlorophyll c, fucoxanthin and violaxanthin in O. luteus, showed a maximum at the onset of light and decreased during the light period. The values of the chlorophyll a:chlorophyll c, chlorophyll a:peridinin and chlorophyll a:fucoxanthin ratios are compared with data reported in the literature.     

PHOTOSYNTHESIS IN THE FEMALE GAMETOPHYTE OF GINKGO BILOBA

The female gametophyte of Ginkgo biloba is the only seed plant gametophyte known to contain chlorophyll. Measurements of photosynthetically active radiation (PAR) indicate that a gametophyte growing within an ovule can receive significant quantities of light (70 μmol photons m^–2 s^–1). Under the conditions of our experiments, whole gametophytes dissected free from ovules were capable of gross photosynthesis, but not net photosynthesis. On a dry wt basis, the maximum rate of carbon fixation under near saturating light intensities was 3.64 × 10^–3 μmol CO₂g^–1 s^–1. The unique ability of the female gametophyte of Ginkgo biloba to produce chlorophyll and engage in photosynthesis appears to result from its exposure to sufficient levels of light and a predisposition to react to this stimulus by the development of a functional photosynthetic apparatus.     

STUDIES ON THE AUTECOLOGY OF THE MARINE DIATOM THALASSIOSIRA NORDENSKIÖLDIICLEVE. 1. THE INFLUENCE OF DAYLENGTH,LIGHT INTENSITY,AND TEMPERATURE ON GROWTH1

The marine diatom Thalassiosira nordenskiöldii Clave was grown at 48 different combinations of daylength (9:15, 12:12, 15:9 LD), light intensity (0.011, 0.027, 0.066, 0.100 ly/min [g cal/cm²/min]), and temperature (0, 5, 10, 15 C). Growth occurred at all combinations of light and temperature except at 15 C at the highest light level. Maximum growth (K = 1.8 doublings/day) occurred at 10 C under the 15:9 LD cycle. At 15 C the maximum rate was 1.7 doublings/day but occurred at the shortest day-length (9:15 LD). The maxima at 5 and 0 C were 1.32 and 0.67 doubling/day, respectively. At 0 C growth was similar over a wide range of light intensities (K = 0.6–0.65), with, maximum growth being attained at a much lower light intensity than at 5 C. Above 5 C there was a decrease in the light intensity at which maximum growth occurred and excessive light became inhibitory to growth. At 15 C the light intensity at which maximum growth occurred was greater with shorter day-lengths. The temperature optimum was 10 C at 15:9 and 15 C at 9:15 LD. The chlorophyll a content of the cells was greatest under low light intensities and short daylengths, while temperature had a variable effect. The response of Thalassiosira in the laboratory contrasts with, its apparent preference for low temperatures in nature (0–5 C). The experiments suggest that the termination of the bloom of Thalassiosira in Narragansett Bay and elsewhere is not solely temperature dependent.     

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 c₂, 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. Low light cells yielded five distinct chlorophyll-protein complexes (I to V), while only four (I′ to IV′) were evident in preparations of high light cells. Both high molecular weight complexes I and I′ were dominated by chlorophyll a absorption and associated with minor amounts of chlorophyll c. Both complexes II and II′ were chlorophyll a-chlorophyll c₂-protein complexes devoid of peridinin and unique to dinoflagellates. The chlorophyll a:c₂ molar ratio of both complexes was 1:3, indicating significant chlorophyll c enrichment over thylakoid membrane chlorophyll a:c ratios of 1.8 to 2:1. Low light complex III differed from all other high or low light complexes in that it possessed peridinin and had a chlorophyll a:c₂ ratio of 1:1. Low light complexes IV and V and high light complexes III′ and IV′ were spectrally similar, had high chlorophyll a:c₂ ratios (4:1), and were associated with peridinin. The effects of growth irradiance on the composition of chlorophyll-protein complexes in Gonyaulax polyedra differed from those described for other chlorophyll c-containing plant species.     

KINETICS OF CHLOROPHYLL a AND PLASTOQUINONE A: CHANGES IN RESPONSE TO LIGHT INTENSITY1

The thermophilic blue-green Synechococcus lividus Y52 was grown at several light intensities between 200 and 2400 ft-c. A 3.4-fold change of cellular chlorophyll (Chl) a content was found. The concentration of Plastoquinone A (PQA) per Chl a varied by a factor of 7, whereas PQA per cell varied by 1.4. Adaptation to light intensity appeared to occur through changes in size of the light-gathering antenna of chlorophyll and phycocyanin while the concentration of PQA remained nearly constant.     

Tuning Energy Transfer in the Peridinin–chlorophyll Complex by Reconstitution with Different Chlorophylls

In vitro studies of the carotenoid peridinin, which is the primary pigment from the peridinin chlorophyll-a protein (PCP) light harvesting complex, showed a strong dependence on the lifetime of the peridinin lowest singlet excited state on solvent polarity. This dependence was attributed to the presence of an intramolecular charge transfer (ICT) state in the peridinin excited state manifold. The ICT state was also suggested to be a crucial factor in efficient peridinin to Chl-a energy transfer in the PCP complex. Here we extend our studies of peridinin dynamics to reconstituted PCP complexes, in which Chl-a was replaced by different chlorophyll species (Chl-b, acetyl Chl-a, Chl-d and BChl-a). Reconstitution of PCP with different Chl species maintains the energy transfer pathways within the complex, but the efficiency depends on the chlorophyll species. In the native PCP complex, the peridinin S₁/ICT state has a lifetime of 2.7 ps, whereas in reconstituted PCP complexes it is 5.9 ps (Chl-b) 2.9 ps (Chl-a), 2.2 ps (acetyl Chl-a), 1.9 ps (Chl-d), and 0.45 ps (BChl-a). Calculation of energy transfer rates using the Förster equation explains the differences in energy transfer efficiency in terms of changing spectral overlap between the peridinin emission and the absorption spectrum of the acceptor. It is proposed that the lowest excited state of peridinin is a strongly coupled S₁/ICT state, which is the energy donor for the major energy transfer channel. The significant ICT character of the S₁/ICT state in PCP enhances the transition dipole moment of the S₁/ICT state, facilitating energy transfer to chlorophyll via the Förster mechanism. In addition to energy transfer via the S₁/ICT, there is also energy transfer via the S₂ and hot S₁/ICT states to chlorophyll in all reconstituted PCP complexes.     

SUN-SHADE ECOTYPES OF A BLUEGREEN ALGA IN A HOT SPRING1,2

Clone cultures of the thermophilic alga Plectonema notatum Schmidle were established from cells collected from the high and low light intensity regions of the algal mat which developed in Jerry Johnson Hot Spring, Idaho. Clones isolated from cither high, or low light intensify zones were grown at light intensities of 8000 and 400 ft-c. The existence of specialized and genetically fixed sun or shade ecotypes was evidenced, by the ability of low light intensity clones to synthesize more light harvesting chlorophyll a when grown under low light conditions than the high light clones. High light clones showed light saturation of photosynthesis at higher light intensities with higher carboxylating enzyme activities and less chlorophyll a than low light clones when both were cultured at 8000 ft-c. These clones displayed infraspecific variation along the light intensity gradient and therefore exist as a mosaic of light, intensity ecotypes.     

A QUALITATIVE AND QUANTITATIVE STUDY OF PLASTOQUINONE A IN TWO THERMOPHILIC BLUE-GREEN ALGAE1

The thermophilic blue-green alga Synechococcus lividus strain Y52 was shown to have plastoquinone (PQ) types A, B, and C. The values PQA per cell and chlorophyll a per cell are presented with the changes in the ratio of PQA to chlorophyll a for cells grown at different light intensities.     

EFFECTS OF VARIATIONS IN LIGHT INTENSITY ON THE EFFICIENCY OF GROWTH OF SKELETONEMA COSTATUM (BACILLARIOPHYCEAE) IN A CYCLOSTAT1

The relative importance of respiration and organic carbon release to the efficiency of carbon specific growth of Skeletonema costatum (Grev.) Clave was evaluated over a light range from 1500–15 μE · m^?2· s^?1. Net growth efficiency ranged from 0.45–0.69 with a maximum at 130 μE · m^?2· s^?1. Respiration was 93% or more of the variations in growth efficiency. Organic carbon release ranged from 0–7% of gross production and increased with light intensity. Carbon specific particulate production was a hyperbolic function of incident light intensity and was related exponentially to particulate carbon production per unit chlorophyll a. Full sunlight conditions, 1500 μE · m^?2· s^?1, did not induce photoinhibition of gross production. Variations in the efficiency of growth of S. costatum were minimized over a wide range of light intensities mainly because of variations in cellular pigments which permitted the efficient utilization of available light energy, and a reduction in the losses of carbon which increases the growth rate, possibly as a consequence of the recycling of respired carbon within the cell.     

Structural Confirmation of a Unique Carotenoid Lactoside,P457, in Symbiodinium sp. Strain nbrc 104787 Isolated from a Sea Anemone and its Distribution in Dinoflagellates and Various Marine Organisms

The molecular structure of the carotenoid lactoside P457, (3S,5R,6R,3′S,5′R,6′S)‐13′‐cis‐5,6‐epoxy‐3′,5′‐dihydroxy‐3‐(β‐d ‐galactosyl‐(1→4)‐β‐d ‐glucosyl)oxy‐6′,7′‐didehydro‐5,6,7,8,5′,6′‐hexahydro‐β,β‐caroten‐20‐al, was confirmed by spectroscopic methods using Symbiodinium sp. strain NBRC 104787 cells isolated from a sea anemone. Among various algae, cyanobacteria, land plants, and marine invertebrates, the distribution of this unique diglycosyl carotenoid was restricted to free‐living peridinin‐containing dinoflagellates and marine invertebrates that harbor peridinin‐containing zooxanthellae. Neoxanthin appeared to be a common precursor for biosynthesis of peridinin and P457, although neoxanthin was not found in peridinin‐containing dinoflagellates. Fucoxanthin‐containing dinoflagellates did not possess peridinin or P457; green dinoflagellates, which contain chlorophyll a and b, did not contain peridinin, fucoxanthin, or P457; and no unicellular algae containing both peridinin and P457, other than peridinin‐containing dinoflagellates, have been observed. Therefore, the biosynthetic pathways for peridinin and P457 may have been coestablished during the evolution of dinoflagellates after the host heterotrophic eukaryotic microorganism formed a symbiotic association with red alga that does not contain peridinin or P457.     

THE EFFECT OF WAVELENGTH AND INTENSITY OF LIGHT ON THE PROPORTION OF PIGMENTS IN PORPHYRIDIUM CRUENTUM

Brody , Marcia , and Robert Emerson . (U. Illinois, Urbana.) The effect of wavelength and intensity of light on the proportion of pigments in Porphyridium cruentum. Amer. Jour. Bot. 46(6): 433–440. Illus. 1959.—We specify a medium for Porphyridium cruentum made from distilled water and inorganic salts, without addition of earth extract, sea water or other supplements of uncertain composition, which sustains excellent growth through successive transfers. By control of intensity and wavelength of light used for growing the cultures, we have found it possible to vary the concentrations of chlorophyll and phycoerythrin, and also the proportions of these 2 pigments, over a considerable range. The changes in proportion of red and green pigments, in response to culturing in green and in blue light, are opposite to the changes that would be predicted from Engelmann's theory of complementary chromatic adaptation, when the intensities of the blue and green light are of the order of 10⁴ ergs/cm²/sec. (or greater). However, at about 1/100 of this intensity, the changes are in the direction of complementary chromatic adaptation.     

Endozoic algae in shelled gastropods — a new symbiotic association in coral reefs?

Live algae were found in the hepatopancreas and gonads of the Red Sea snail Strombus tricornis. These organs are constantly concealed within the upper whorls of the snail's shell. Light penetration was 5–15% of the incident light reaching the shell. Pigment analysis indicated the presence of chlorophyll a, c and peridinin, a composition resembling the Dinoflagellata. Chlorophyll a concentration in the algae was 1.18±0.36 pg chl/cell. ¹⁴C assimilation of isolated algae incubated in the light exceeded that of dark controls, demonstrating the photosynthetic activity of the endozoic algae.     

CAROTENOID PIGMENTS OF THE DINOFLAGELLATE GLENODINIUM FOLIACEUM STEIN12

The marine dinoflagellate, Glenoclinium foliaceum Stein, has been shown to contain fucoxanthin, instead of peridinin, as the major xanthophyll. In addition, 2 carotenes–β-carotene and a compound with spectral properties reminiscent of isomerized y-carotene–and 2 xanthophylls–diadinoxanthin and an unidentified compound–were also isolated. These results support an earlier work that indicated the possible presence of fucoxanthin in some members of the Pyrrophyta.     

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

Summary Whole cell absorption curves of the marine dinoflagellate Glenodinium sp., cultured at irradiances of 250W/cm² (low light) and 2500W/cm² (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 difference spectrum supported the view that increased pigmentation, resulting from low light conditions, was largely due to an increase in cell content of a peridinin-chlorophyll a-protein (PCP) and an unidentified chlorophyll a component of the chloroplast membrane. Photosynthetic action spectrum measurements indicated that chlorophyll a, peridinin, and very likely chlorophyll c, were effective light-harvesting pigments for photosynthesis in both high and low light grown cultures of Glenodinium sp. Comparison of action spectra and absorption spectra suggested that low light grown cells selectively increased cellular absorption in the 480 nm to 560 nm region, and effectively utilized this spectral region for the promotion of oxygen evolution.Abbreviations PCP peridinin-chlorophyll a-protein - SIO (F.T. Haxo) Scripps Institution of Oceanography collection     

THE FINE STRUCTURE OF RHODOSPIRILLUM RUBRUM

The fine structure of Rhodospirillum rubrum grown under a series of defined conditions has been examined in thin sections prepared by the methods of Ryter and Kellenberger. In cells grown anaerobically at different light intensities, the abundance of 500 A membrane-bounded vesicles in the cytoplasm is inversely related to light intensity, and directly related to cellular chlorophyll content. When the chlorophyll content of the cell is low, the vesicles are exclusively peripheral in location; they extend more deeply into the cytoplasm when the chlorophyll content is high. Typical vesicles also occur, though rarely, in cells grown aerobically in the dark, which have a negligible chlorophyll content. When synthesis of the photosynthetic pigment system is induced in a population of aerobically grown cells by incubation under semianaerobic conditions in the dark, the vesicles become increasingly abundant with increasing cellular chlorophyll content, and the cells eventually acquire the cytoplasmic structure that is characteristic of cells growing anaerobically at a high light intensity. Poststaining with lead hydroxide reveals that the membranes surrounding the 500 A vesicles are indistinguishable in structure from the cytoplasmic membrane, and continuous with it in some areas of the sections. The bearing of these observations on current notions concerning the organization of the bacterial photosynthetic apparatus is discussed.     

PIGMENTS,FATTY ACIDS,AND STEROLS OF THE TOXIC DINOFLAGELLATE GYMNODINIUM CATENATUM1

Cultures and field samples of the toxic dinoflagellate Gymnodinium catenatum Graham from Tasmania, Australia, were analyzed for pigment, fatty acid, and sterol composition. Gymnodinium catenatum contained the characteristic pigments of photosynthetic dinoflagellates, including chlorophyll a, chlorophyll c₂, and the carotenoids peridinin, dinoxanthin, diadinoxanthin, diatoxanthin, and β,β-carotene. In midlogarithmic and early stationary phase cultures, the chlorophyll a content ranged 50–72 pg · cell^?1, total lipids 956–2084 pg · cell^?1, total fatty acids 426–804 pg · cell^?1, and total sterols 8–20 pg · cell^?1. The major fatty acids (in order of decreasing abundance) were 16:0, 22:6(n-3), and 20:5(n-3) (collectively 65–70% of the total fatty acids), followed by 16:1(n-7), 18:2(n-6), and 14:0. This distribution is characteristic of most dinoflagellates, except for the low abundance (<3%) of the fatty acid 18:5(n-3), considered by some authors to be a marker for dinoflagellates. The three major sterols were 4α-methyl-5α-cholest-7-en-3β-ol, 4α,23,24-trimethyl-5α-cholest-22E-en-3β-ol (the dinoflagellate sterol, dinosterol), and 4α,23,24-trimethyl-5α-cholest-7-en-3β-ol. These three sterols comprised about 75% of the total sterols in both logarithmic and early stationary phase cultures, and they were also found in high proportions (22–25%) in natural dinoflagellate bloom samples. 4-Desmethyl sterols, which are common in most microalgae, were only present in trace amounts in G. catenatum. The chemotaxonomic affinities of G. catenatum and the potential for using specific signature lipids for monitoring toxic dinoflagellate blooms are discussed.     

THE TIME-COURSE OF PHOTOADAPTATION TO LOW-LIGHT IN PROROCENTRUM MARIAE-LEBOURIAE (DINOPHYCEAE)1

The estuarine dinoflagellate, Prorocentrum mariaelebouriae (Parke & Ballantine 1957) Faust 1974 undergoes increases in pigmentation and photosynthetic efficiency within several days of downward light shifts. These changes can be described by first-order kinetics, as has been reported previously for Chlorophyll (Chl) a in several phytoplankton species. The studies described in this paper were conducted with isolates of populations of Prorocentrum from the Chesapeake Bay. We determined rates of adaptation to low-light for cultures grown at a range of photon flux densities (I₀= 2.65–26.2 E.m^?2, d^?1, shifted to 6.3–7.0% I₀) at three temperatures (10°, 15°, and 20° C), bracketing the conditions this species experiences in situ. In this paper, I report the time-course of changes in α, P_max Chl a, peridinin, and I_k and first-order rate constants, K₁ for changes in α, Chl a and peridinin. cell^?1. K₁ for changes in α cell^?1 averaged 1.58 × 10^?2 h^?1 for conditions encompassing five light treatments and three temperatures; the corresponding mean for Chl a was 1.59 × 10^?2 h^?1. Increases in peridinin measured for five light treatments at 15° C showed a mean K₁ of 1.22 × 10^?2 h^?1, Average percent changes in per cell α, Chl a, and peridinin ranged from 0.4–4.0% h^?1 (10–90% d^?1) following exposure to low-light. Photoadaptive changes are important to Prorocentrum because in nature it occupies turbid waters (K_t≥ 0.5 m^?1) where the mixing depth often exceeds the depth of the photic layer. Cells are entrained beneath a seasonally-stable density discontinuity and are exposed to very low-light (< I E.m^?2.d^?1) for days to weeks during subpycnocline transport. The ability of this species to undergo changes in pigmentation and photosynthetic physiology confers increased efficiency of light harvesting and contributes to this species’survival in the estuary where it is an important component of the dinoflagellate flora.     

HETEROTROPHIC GROWTH OF ULVA LACTUCA (CHLOROPHYCEAE)1

The effect of external glucose (51 mM) and acetate (13 mM) on growth and photosynthetic capacity of Ulva lactuca L. was tested in laboratory cultures over 41 days in the dark and in dim light (0.9 μmol photons·m^?2·s^?1) at 7–8° C. Glucose and acetate had a significant positive effect on growth rate, chlorophyll content, and quantum yield for discs grown in the dark and in dim light. The carbon gain from heterotrophic uptake was low and only allowed U. lactuca to maintain a specific uptake was low and only allowed U. lactuca to maintain a specific growth rate of 0.005 day^?1 compared to 0.06–0.1 day^?1 at higher light intensities. However, plants with added organic substrate maintained a normal chlorophyll content and were able to photosynthesize whereas control plants lost pigmentation and photosynthetic capability after 41 days in both dim light and darkness, probably because of disorganization of the photosynthetic apparatus. This suggest that the ecological significance of heterotrophic uptake is to allow U. lactuca to survive during prolonged low light conditions with an intact photosynthetic apparatus.