Photosystem electron-transport capacity and light-harvesting antenna size in maize chloroplasts

Spectrophotometric and kinetic measurements were applied to yield photosystem (PS) stoichiometries and the functional antenna size of PSI, PSII_α, and PSII_β in Zea mays chloroplasts in situ. Concentrations of PSII and PSI reaction centers were determined from the amplitude of the light-induced absorbance change at 320 and 700 nm, which reflect the photoreduction of the primary electron acceptor Q of PSII and the photooxidation of the reaction center P700 of PSI, respectively. Determination of the functional chlorophyll antenna size (N) for each photosystem was obtained from the measurement of the rate of light absorption by the respective reaction center. Under the experimental conditions employed, the rate of light absorption by each reaction center was directly proportional to the number of light-harvesting chlorophyll molecules associated with the respective photosystem. We determined N_P700 = 195, N_α = 230, N_β = 50 for the number of chlorophyll molecules in the light-harvesting antenna of PSI, PSII_α, and PSII_β, respectively. The above values were used to estimate the PSII/PSI electron-transport capacity ratio (C) in maize chloroplasts. In mesophyll chloroplasts C > 1.4, indicating that, under green actinic excitation when Chl a and Chl b molecules absorb nearly equal amounts of excitation, PSII has a capacity to turn over electrons faster than PSI. In bundle sheath chloroplasts C < 1, suggesting that such chloroplasts are not optimally poised for linear electron transport and reductant generation.     

Activation of a Reserve Pool of Photosystem II in Chlamydomonas reinhardtii Counteracts Photoinhibition

The effect of strong irradiance (2000 micromole photons per square meter per second) on PSII heterogeneity in intact cells of Chlamydomonas reinhardtii was investigated. Low light (LL, 15 micromole photons per square meter per second) grown C. reinhardtii are photoinhibited upon exposure to strong irradiance, and the loss of photosynthetic functioning is due to damage to PSII. Under physiological growth conditions, PSII is distributed into two pools. The large antenna size (PSII_α) centers account for about 70% of all PSII in the thylakoid membrane and are responsible for plastoquinone reduction (Q_b-reducing centers). The smaller antenna (PSII_β) account for the remainder of PSII and exist in a state not yet able to photoreduce plastoquinone (Q_b-nonreducing centers). The exposure of C. reinhardtii cells to 60 minutes of strong irradiance disabled about half of the primary charge separation between P680 and pheophytin. The PSII_β content remained the same or slightly increased during strong-irradiance treatment, whereas the photochemical activity of PSII_α decreased by 80%. Analysis of fluorescence induction transients displayed by intact cells indicated that strong irradiance led to a conversion of PSII_β from a Q_b-nonreducing to a Q_b-reducing state. Parallel measurements of the rate of oxygen evolution revealed that photosynthetic electron transport was maintained at high rates, despite the loss of activity by a majority of PSII_α. The results suggest that PSII_β in C. reinhardtii may serve as a reserve pool of PSII that augments photosynthetic electron-transport rates during exposure to strong irradiance and partially compensates for the adverse effect of photoinhibition on PSII_α.     

On the Factors Which Determine Massive beta-Carotene Accumulation in the Halotolerant Alga Dunaliella bardawil

Dunaliella bardawil, a β-carotene-accumulating halotolerant alga, has been analyzed for the effect of various growth conditions on its pigment content, and compared with Dunaliella salina, a β-carotene nonaccumulating species. In D. bardawil, increasing light intensity and light period or inhibiting growth by various stress conditions such as nutrient deficiency or high salt concentration caused a decrease in the content of chlorophyll per cell and an increase in the amount of β-carotene per cell. As a result, the β-carotene-to-chlorophyll ratio increased from about 0.4 to 13 grams per gram and the alga changed its visual appearance from green to deep orange. D. salina grown similarly decreased in content of both chlorophyll and β-carotene per cell and the culture turned from green to yellowish. Low chlorophyll-containing cells of D. bardawil or D. salina exhibit very high photosynthetic rates when expressed on a chlorophyll basis (~600 micromoles O₂ evolved per milligram chlorophyll per hour).

Variation of pigment content in D. bardawil by a large variety of environmental agents has been correlated with the integral irradiance received by the algal culture during a division cycle. The higher the integral irradiance per division cycle, the lower the chlorophyll content per cell; the higher the β-carotene content per cell, and therefore the higher the β-carotene-to-chlorophyll ratio. The results are interpreted as indicating a protecting effect of β-carotene against injury by high irradiance under conditions of impairment in chlorophyll content per cell.

     

Antenna Sizes of Photosystem I and Photosystem II in Chlorophyll b-Deficient Mutants of Rice. Evidence for an Antenna Function of Photosystem II Centers That are Inactive in Electron Transport

Light-harvesting capacities of photosystem I (PSI) and photosystemII (PSII) in a wild-type and three chlorophyll b-deficient mutantstrains of rice were determined by measuring the initial slopeof light-response curve of PSI and PSII electron transport andkinetics of light-induced redox changes of P-700 and Q_A, respectively.The light-harvesting capacity of PSI determined by the two methodswas only moderately reduced by chlorophyll b-deficiency. Analysisof the fluorescence induction that monitors time course of Q_Aphotoreduction showed that both relative abundance and antennasize of PSII_a decrease with increasing deficiency of chlorophyllb and there is only PSII_rß in chlorina 2 which totallylacks chlorophyll b. The numbers of antenna chlorophyll moleculesassociated with the mutant PSII centers were, therefore, threeto five times smaller than that of PSII_a in the wild type rice.Rates of PSII electron transport determined on the basis ofPSII centers in the three mutants were 60–70% of thatin the normal plant at all photon flux densities examined, indicatingthat substantial portions of the mutant PSII centers are inactivein electron transport. The initial slopes of light-responsecurves of PSII electron transport revealed that the functionalantenna sizes of the active populations of PSII centers in themutants correspond to about half that of PSII in the wild typerice. Thus, the numbers of chlorophyll molecules that serveas antenna of the oxygen-evolving PSII centers in the mutantsare significantly larger than those that are actually associatedwith each PSII center. It is proposed that the inactive PSIIserves as an antenna of the active PSII in the three chlorophyllb-deficient mutants of rice. In spite of the reduced antennasize of PSII, therefore, the total light-harvesting capacityof PSII approximately matches that of PSI in the mutants. (Received July 29, 1994; Accepted February 7, 1996)     

Growth under Red Light Enhances Photosystem II Relative to Photosystem I and Phycobilisomes in the Red Alga Porphyridium cruentum

Acclimation of the photosynthetic apparatus to light absorbed primarily by photosystem I (PSI) or by photosystem II (PSII) was studied in the unicellular red alga Porphyridium cruentum (ATCC 50161). Cultures grown under green light of 15 microeinsteins per square meter per second (PSII light; absorbed predominantly by the phycobilisomes) exhibited a PSII/PSI ratio of 0.26 ± 0.05. Under red light (PSI light; absorbed primarily by chlorophyll) of comparable quantum flux, cells contained nearly five times as many PSII per PSI (1.21 ± 0.10), and three times as many PSII per cell. About 12% of the chlorophyll was attributed to PSII in green light, 22% in white light, and 39% in red light-grown cultures. Chlorophyll antenna sizes appeared to remain constant at about 75 chlorophyll per PSII and 140 per PSI. Spectral quality had little effect on cell content or composition of the phycobilisomes, thus the number of PSII per phycobilisome was substantially greater in red light-grown cultures (4.2 ± 0.6) than in those grown under green (1.6 ± 0.3) or white light (2.9 ± 0.1). Total photosystems (PSI + PSII) per phycobilisome remained at about eight in each case. Carotenoid content and composition was little affected by the spectral composition of the growth light. Zeaxanthin comprised more than 50% (mole/mole), β-carotene about 40%, and cryptoxanthin about 4% of the carotenoid pigment. Despite marked changes in the light-harvesting apparatus, red and green light-grown cultures have generation times equal to that of cultures grown under white light of only one-third the quantum flux.     

Stoichiometry of Photosystem I, Photosystem II, and Phycobilisomes in the Red Alga Porphyridium cruentum as a Function of Growth Irradiance

Cells of the red alga Porphyridium cruentum (ATCC 50161) exposed to increasing growth irradiance exhibited up to a three-fold reduction in photosystems I and II (PSI and PSII) and phycobilisomes but little change in the relative numbers of these components. Batch cultures of P. cruentum were grown under four photon flux densities of continuous white light; 6 (low light, LL), 35 (medium light, ML), 180 (high light, HL), and 280 (very high light, VHL) microeinsteins per square meter per second and sampled in the exponential phase of growth. Ratios of PSII to PSI ranged between 0.43 and 0.54. About three PSII centers per phycobilisome were found, regardless of growth irradiance. The phycoerythrin content of phycobilisomes decreased by about 25% for HL and VHL compared to LL and ML cultures. The unit sizes of PSI (chlorophyll/P₇₀₀) and PSII (chlorophyll/Q_A) decreased by about 20% with increase in photon flux density from 6 to 280 microeinsteins per square meter per second. A threefold reduction in cell content of chlorophyll at the higher photon flux densities was accompanied by a twofold reduction in β-carotene, and a drastic reduction in thylakoid membrane area. Cell content of zeaxanthin, the major carotenoid in P. cruentum, did not vary with growth irradiance, suggesting a role other than light-harvesting. HL cultures had a growth rate twice that of ML, eight times that of LL, and slightly greater than that of VHL cultures. Cell volume increased threefold from LL to VHL, but volume of the single chloroplast did not change. From this study it is evident that a relatively fixed stoichiometry of PSI, PSII, and phycobilisomes is maintained in the photosynthetic apparatus of this red alga over a wide range of growth irradiance.     

Compensatory Alterations in the Photochemical Apparatus of a Photoregulatory, Chlorophyll b-Deficient Mutant of Maize

Characterization of the functional organization of the photochemical apparatus in the light sensitive chlorophyll b-deficient oil yellow-yellow green (OY-YG) mutant of maize (Zea mays) is presented. Spectrophotometric and kinetic analysis revealed substantially lower amounts of the light harvesting complex of photosystem II (LHCII-peripheral) in high light-grown OY-YG thylakoids. However, accumulation of a tightly bound LHCII appears unaffected by the lesion. Changes in photosystem (PS) stoichiometry include lower amounts of PSII with characteristic fast kinetics (PSII_α) and a substantial accumulation of PSII centers with characteristic slow kinetics (PSII_β) in the thylakoid membrane of the OY-YG mutant. Thus, PSII_β is the dominant photosystem in the mutant chloroplasts. In contrast to wild type, roughly 80% of the mutant PSII_β centers are functionally coupled to the plastoquinone pool and are probably localized in the appressed regions of the thylakoid membrane. These centers, designated PSII_β-Q_B-reducing (Q_B being the secondary electron quinone acceptor of PSII), are clearly distinct from the typical PSII_β-Q_B-nonreducing centers found in the stroma lamellae of wild-type chloroplasts. It is concluded that the observed changes in the stoichiometry of electron-transport complexes reflect the existence of a regulatory mechanism for the adjustment of photosystem stoichiometry in chloroplasts designed to correct any imbalance in light absorption by the two photosystems.     

Characterization of Photosystem II Activity and Heterogeneity during the Cell Cycle of the Green Alga Scenedesmus quadricauda

The photosynthetic activity of the green alga Scenedesmus quadricauda was investigated during synchronous growth in light/dark cycles. The rate of O₂ evolution increased 2-fold during the first 3 to 4 h of the light period, remained high for the next 3 to 4 h, and then declined during the last half of the light period. During cell division, which occurred at the beginning of the dark period, the ability of the cells to evolve O₂ was at a minimum. To determine if photosystem II (PSII) controls the photosynthetic capacity of the cells during the cell cycle we measured PSII activity and heterogeneity. Measurements of electron-transport activity revealed two populations of PSII, active centers that contribute to carbon reduction and inactive centers that do not. Measurements of PSII antenna sizes also revealed two populations, PSII_α and PSII_β, which differ from one another by their antenna size. During the early light period the photosynthetic capacity of the cells doubled, the O₂-evolving capacity of PSII was nearly constant, the proportion of PSII_β centers decreased to nearly zero, and the proportion of inactive PSII centers remained constant. During the period of minimum photosynthetic activity 30% of the PSII centers were insensitive to the inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea, which may be related to reorganization of the thylakoid membrane. We conclude from these results that PSII does not limit the photosynthetic activity of the cells during the first half of the light period. However, the decline in photosynthetic activity observed during the last half of the light period can be accounted for by limited PSII activity.     

Distribution of Thylakoid Proteins between Stromal and Granal Lamellae in Spirodela : Dual Location of Photosystem II Components

We have quantified the lateral distribution of 12 thylakoid proteins of Spirodela oligorrhiza by immunoblot analysis of detergent-derived granal and stromal lamellae. The immunological, ultrastructural, cytochemical, and biophysical measurements each indicated the expected overall separation of photosystem II (PSII) and photosystem I (PSI) components; however, certain proteins were not completely localized to one lamellar fraction. The apoproteins of the light harvesting chlorophyll a/b complex, subunit 1 of PSI and the components of the PSII reaction center (the 32 kilodalton, D2, and cytochrome b₅₅₉ proteins) were dually located between granal and stromal lamellae. Proteins associated exclusively with one of the membrane types were: in granal lamellae, the 43 and 51 kilodalton PSII proteins, and in stromal lamellae, the α and β subunits of the proton ATPase.     

Photoacclimation in the Red Alga Porphyridium cruentum: Changes in Photosynthetic Enzymes, Electron Carriers, and Light-Saturated Rate of Photosynthesis as a Function of Irradiance and Spectral Quality

Acclimation of the photosynthetic apparatus to changes in the light environment was studied in the unicellular red alga Porphyridium cruentum (American Type Culture Collection No. 50161). Absolute or relative amounts of four photosynthetic enzymes and electron carriers were measured, and the data were compared with earlier observations on light-harvesting components (F.X. Cunningham, Jr., R.J. Dennenberg, L. Mustárdy, P.A. Jursinic, E. Gantt [1989] Plant Physiol 91: 1179-1187; F.X. Cunningham, Jr., R.J. Dennenberg, P.A. Jursinic, E. Gantt [1990] Plant Physiol 93: 888-895) and with measurements of photosynthetic capacity. P_max, the light-saturated rate of photosynthesis on a chlorophyll (Chl) basis, increased more than 4-fold with increase in growth irradiance from 6 to 280 μeinsteins·m⁻²·s⁻¹. Amounts of ferredoxin-NADP⁺ reductase, ribulose-1,5-bisphosphate carboxylase, and cytochrome f increased in parallel with P_max, whereas numbers of the light-harvesting complexes (photosystem [PS] I, PSII, and phycobilisomes) changed little, and ATP synthase increased 7-fold relative to Chl. The calculated minimal turnover time for PSII under the highest irradiance, 5 ms, was thus about 4-fold faster than that calculated for cultures grown under the lowest irradiance (19 ms). A change in the spectral composition of the growth light (irradiance kept constant at 15 μeinsteins·m⁻²·s⁻¹) from green (absorbed predominantly by the phycobilisome antenna of PSII) to red (absorbed primarily by the Chl antenna of PSI) had little effect on the amounts of ribulose-1,5-bisphosphate carboxylase, ATP synthase, and phycobilisomes on a Chl, protein, or thylakoid area basis. However, the number of PSI centers declined by 40%, cytochrome f increased by 40%, and both PSII and ferredoxin-NADP⁺ reductase increased approximately 3-fold on a thylakoid area basis. The substantial increase in ferredoxin-NADP⁺ reductase under PSI light is inconsistent with a PSI-mediated reduction of NADP as the sole function of this enzyme. Our results demonstrate a high degree of plasticity in content and composition of thylakoid membranes of P. cruentum.     

Association of chlorophyll a/b-binding Pcb proteins with photosystems I and II in Prochlorothrix hollandica

Action spectra for photosystem II (PSII)-driven oxygen evolution and of photosystem I (PSI)-mediated H₂ photoproduction and photoinhibition of respiration were used to determine the participation of chlorophyll (Chl) a/b-binding Pcb proteins in the functions of pigment apparatus of Prochlorothrix hollandica. Comparison of the in situ action spectra with absorption spectra of PSII and PSI complexes isolated from the cyanobacterium Synechocystis 6803 revealed a shoulder at 650 nm that indicated presence of Chl b in the both photosystems of P. hollandica. Fitting of two action spectra to absorption spectrum of the cells showed a chlorophyll ratio of 4:1 in favor of PSI. Effective antenna sizes estimated from photochemical cross-sections of the relevant photoreactions were found to be 192 ± 28 and 139 ± 15 chlorophyll molecules for the competent PSI and PSII reaction centers, respectively. The value for PSI is in a quite good agreement with previous electron microscopy data for isolated Pcb-PSI supercomplexes from P. hollandica that show a trimeric PSI core surrounded by a ring of 18 Pcb subunits. The antenna size of PSII implies that the PSII core dimers are associated with ∼ 14 Pcb light-harvesting proteins, and form the largest known Pcb-PSII supercomplexes.     

Dunaliella salina (Chlorophyta) with small chlorophyll antenna sizes exhibit higher photosynthetic productivities and photon use efficiencies than normally pigmented cells

The photon use efficiencies and maximal rates of photosynthesis in Dunaliella salina (Chlorophyta) cultures acclimated to different light intensities were investigated. Batch cultures were grown to the mid-exponential phase under continuous low-light (LL: 100 μmol photon m^-2 s^-1) or high-light (HL: 2000 μmol photon m^-2 s^-1) conditions. Under LL, cells were normally pigmented (deep green) containing ∼500 chlorophyll (Chl) molecules per photosystem II (PSII) unit and ∼250 Chl molecules per photosystem I (PSI). HL-grown cells were yellow-green, contained only 60 Chl per PSII and 100 Chl per PSI and showed signs of chronic photoinhibition, i.e., accumulation of photodamaged PSII reaction centers in the chloroplast thylakoids. In LL-grown cells, photosynthesis saturated at ∼200 μmol photon m^-2 s^-1 with a rate (P_max) of ∼100 mmol O₂ (mol Chl)^-1 s^-1. In HL-grown cells, photosynthesis saturated at much higher light intensities, i.e. ∼2500 μmol photon m^-2 s^-1, and exhibited a three-fold higher P_max (∼300 mmol O₂ (mol Chl)^-1 s^-1) than the normally pigmented LL-grown cells. Recovery of the HL-grown cells from photoinhibition, occurring prior to a light-harvesting Chl antenna size increase, enhanced P_max to ∼675 mmol O₂ (mol Chl)^-1 s^-1. Extrapolation of these results to outdoor mass culture conditions suggested that algal strains with small Chl antenna size could exhibit 2–3 times higher productivities than currently achieved with normally pigmented cells. This revised version was published online in August 2006 with corrections to the Cover Date.     

Rearrangement of the chloroplast thylakoid at chilling temperature in the light

The leaves of chilling-sensitive pumpkin (Cucurbita pepo L.) showed symptoms reminiscent of photoinhibition when kept for 4 days at 5°C in moderate light. A decrease was observed in the variable part of chlorophyll α fluorescence, apparent quantum yield, and maximum rate of O₂ evolution. Chloroplast whole-chain electron transport activity measured from chloroplast thylakoids had decreased to 51% of the control value. Photosystem II (PSII) activity decreased by only 9%, suggesting that photoinhibition was not responsible for the loss of electron transport activity. An increase in the proportion of PSII_β (measured as a β_max value) was observed after the chilling treatment. Fractionation of thylakoid membranes showed a 42% increase in PSII activity in the nonappressed region while that in the appressed region decreased slightly. This was accompanied by a decrease in the ratio of the length of appressed to nonappressed thylakoid membranes. Leaf photosynthesis largely recovered within 24 hours of returning to the original growth conditions. We suggest that the increase in the proportion of PSII_β during chilling in light plays a role in protecting PSII from photoinhibitory damage.     

Photochemical Apparatus Organization in the Chloroplasts of Two Beta vulgaris Genotypes

The composition and structural organization of thylakoid membranes of a low chlorophyll mutant of Beta vulgaris was investigated using spectroscopic, kinetic and electrophoretic techniques. The data obtained were compared with those of a standard F1 hybrid of the same species. The mutant was depleted in chlorophyll b relative to the hybrid and it had a higher photosystem II/photosystem I reaction center (Q/P₇₀₀) ratio and a smaller functional chlorophyll antenna size. Analysis of thylakoid membranes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the mutant lacked a portion of the chlorophyll a/b light-harvesting complex but was enriched in the photosystem II reaction center chlorophyll protein complex. Comparison of functional antenna sizes and of photosystem stoichiometries determined electrophoretically were in good agreement with those determined spectroscopically. Both approaches indicated that about 30% of the total chlorophyll was associated with photosystem I and about 70% with photosystem II. A greater proportion of photosystem II_β was detected in the mutant. The results suggest that a higher photosystem II to photosystem I ratio in the sugar beet mutant has apparently compensated for the smaller photosystem II chlorophyll light-harvesting antenna in its chloroplasts. Moreover, a lack of chlorophyll a/b light-harvesting complex correlates with the abundance of photosystem II_β. It is proposed that a developmental relationship exists between the two types of photosystem II where photosystem II_β is a precursor form of photosystem II_α occurring prior to the addition of the chlorophyll a/b light-harvesting complex and grana formation.     

Differential Control of Xanthophylls and Light-Induced Stress Proteins, as Opposed to Light-Harvesting Chlorophyll a/b Proteins, during Photosynthetic Acclimation of Barley Leaves to Light Irradiance

Barley (Hordeum vulgare L.) plants were grown at different photon flux densities ranging from 100 to 1800 μmol m⁻² s⁻¹ in air and/or in atmospheres with reduced levels of O₂ and CO₂. Low O₂ and CO₂ partial pressures allowed plants to grow under high photosystem II (PSII) excitation pressure, estimated in vivo by chlorophyll fluorescence measurements, at moderate photon flux densities. The xanthophyll-cycle pigments, the early light-inducible proteins, and their mRNA accumulated with increasing PSII excitation pressure irrespective of the way high excitation pressure was obtained (high-light irradiance or decreased CO₂ and O₂ availability). These findings indicate that the reduction state of electron transport chain components could be involved in light sensing for the regulation of nuclear-encoded chloroplast gene expression. In contrast, no correlation was found between the reduction state of PSII and various indicators of the PSII light-harvesting system, such as the chlorophyll a-to-b ratio, the abundance of the major pigment-protein complex of PSII (LHCII), the mRNA level of LHCII, the light-saturation curve of O₂ evolution, and the induced chlorophyll-fluorescence rise. We conclude that the chlorophyll antenna size of PSII is not governed by the redox state of PSII in higher plants and, consequently, regulation of early light-inducible protein synthesis is different from that of LHCII.     

Effects of growth irradiance levels on the ratio of reaction centers in two species of marine phytoplankton

Cells of two species of single-celled marine algae, the diatom Skeletonema costatum (Greve), Cleve, and the chlorophyte Dunaliella tertiolecta Butcher, were cultured in white light of high (500-600 microeinsteins per square meter per second) and low (30 microeinsteins per square meter per second) intensity. For both algal species, cells grown at low light levels contained more chlorophyll a and had a lower ratio of chlorophyll a to chlorophylls b or c than did cells grown at high light levels. When photosynthetic unit sizes were measured on the basis of either oxygen flash yields or P₇₀₀ photooxidation, different results were obtained with the different species. In the chlorophyte, the cellular content of photosystem I (PSI) and photosystem II (PSII) reaction centers increased in tandem as chlorophyll a content increased so that photosynthetic unit sizes changed only slightly and the ratio PSI:PSII reaction centers remained constant at about 1.1. In the diatom, as the chlorophyll content of the cells increased, the number of PSI reaction centers decreased and the number of PSII reaction centers increased so that the ratio of PSI:PSII reaction centers decreased from about unity to 0.44. In neither organism did photosynthetic capacity correlate with changes in cellular content of PSI or PSII reaction centers. The results are discussed in relationship to the physical and biological significance of the photosynthetic unit concept.     

ALGAL PHOTOSYNTHETIC MEMBRANE COMPLEXES AND THE PHOTOSYNTHESIS-IRRADIANCE CURVE: A COMPARISON OF LIGHT-ADAPTATION RESPONSES IN CHLAMYDOMONAS REINHARDTII (CHLOROPHYTA)1

Chlamydomonas reinhardtii was grown at photon flux densities (PFDs) ranging from 47 to 400 μE.m^-2 s^-1. The total cellular content of chlorophyll (Chl) was twice as high in the low light (LL) versus high light (HL) grown cells. On an equal Chl basis, photosystem II (PSII) and cytochrome f (Cyt f) content was higher in HL cells, but photosystem I (PSI) concentration displayed little variation with the light intensity during cell growth. Consequently, there was a shift in the ratio of PSII / PSI and Cyt / PSI from near unity in LL cells to greater than two in HL cells. The functional Chl antenna size of PSII and PSI ranged from 460 and 170 Chl (a + b)in HL-grown cells to 620 and 370 Chl (a+ b)in LL-grown cells, respectively. The initial slope of the Chl-specific photosyn-thesis-irradiance (P-I) curve was similar in LL- and HL-grown cells, but the light saturated rate of photosynthesis was lower under LL. The response to low light was beneficial at the cellular level, since there was an enhancement of photosynthesis in LL. The PFD for the onset of light saturation, 1 was a factor of 2 lower in LL- relative to HL-grown photosythetic membranes. Since growth PFD varied by a factor of ten, photosynthesis shifted from being light-limited in the LL regime to light-saturated in the HL regime. The requirement for balanced absorption of light by the two photosystems constrains the PSII / PSI ratio to near unity when growth is light-limited, but such a constraint does not apply in HL conditions. Instead the concentration of individual electron transport complexes way be related to the pool size necessary for maximum rates of steady-state electron transport. Thus the stoichiometry of electron transport complexes changes in response to growth PFD and this change is correlated with the response flexlbility of algal photosynthesis in diverse light environments.     

Contrasting patterns of photosynthetic acclimation to the light environment are dependent on the differential expression of the responses to altered irradiance and spectral quality

Chl, chlorophyll
Chl a/b, ratio of chlorophyll a to chlorophyll b
Cyt f, cytochrome f
FR, far-red light
LFR, low irradiance, far-red enriched growth light
LHCII, light harvesting complex associated with PSII
LW, low irradiance, white growth light
MW, moderate irradiance, white growth light
PAR, photosynthetically active radiation
P_max, light and CO₂ saturated photosynthetic rate
PSI, photosystem I
PSII, photosystem II

Four plant species (Chamerion angustifolium, Digitalis purpurea, Brachypodium sylvaticum and Plantago lanceolata) which have previously been shown to demonstrate contrasting photosynthetic acclimatory responses to the light environment ( 33 , Plant, Cell and Environment 20, pp. 438–448) were analysed at a biochemical level. Plants were grown under low irradiance with a shade-type spectrum (LFR: 50μmol quanta m^–2 s^–1), moderately high white light (MW: 300μmol quanta m^–2 s^–1) and low irradiance white light (LW: 50μmol quanta m^–2 s^–1). The effects of light quality upon chlorophyll content and photosynthetic capacity were found to be species-dependent. A far-red dependent reduction in chlorophyll was found in three species, and an irradiance-dependent reduction was found in B. sylvaticum, which showed the greatest alteration in the xanthophyll cycle pool size of all species tested under these conditions. Chlorophyll a/b ratios were sensitive to both light quality and quantity in C. angustifolium and D. purpurea, being highest in MW, lowest in LFR, and intermediate in LW, whilst the other species showed no response. Ratios of photosystem II to photosystem I (PSII and PSI) demonstrated a strong irradiance-associated increase in all species except B. sylvaticum, whereas an increase in PSII/PSI in LFR compared to LW conditions was present in all species. A change in chlorophyll a/b was not always associated with a change in PSII/PSI, suggesting that the level of LHCII associated with each PSII varied in some species. Cytochrome f content showed an irradiance-dependent effect only, indicating a relationship with the capacity of electron transport. It is concluded that differing strategies of acclimation to the light environment demonstrated by these species results from differing strengths of expression of a series of independently regulated changes in the levels of photosynthetic components.     

Purification and characterization of photosystem I complex from Synechocystis sp. PCC 6803 by expressing histidine-tagged subunits

We generated Synechocystis sp. PCC 6803 strains, designated F-His and J-His, which express histidine-tagged PsaF and PsaJ subunits, respectively, for simple purification of the photosystem I (PSI) complex. Six histidine residues were genetically added to the C-terminus of the PsaF subunit in F-His cells and the N-terminus of the PsaJ subunit in J-His cells. The histidine residues introduced had no apparent effect on photoautotrophic growth of the cells or the activity of PSI and PSII in thylakoid membranes. PSI complexes could be simply purified from the F-His and J-His cells by Ni²⁺-affinity column chromatography. When thylakoid membranes corresponding to 20 mg chlorophyll were used, PSI complexes corresponding to about 7 mg chlorophyll could be purified in both strains. The purified PSI complexes could be separated into monomers and trimers by ultracentrifugation in glycerol density gradient and high activity was recorded for trimers isolated from the F-His and J-His strains. Blue-Native PAGE and SDS-PAGE analysis of monomers and trimers indicated the existence of two distinct monomers with different subunit compositions and no contamination of PSI with other complexes, such as PSII and Cyt b₆f. Further analysis of proteins and lipids in the purified PSI indicated the presence of novel proteins in the monomers and about six lipid molecules per monomer unit in the trimers. These results demonstrate that active PSI complexes can be simply purified from the constructed strains and the strains are very useful tools for analysis of PSI.     

The Nature of Light-Induced Inhibition of Photosystem II in Pumpkin (Cucurbita pepo L.) Leaves Depends on Temperature

Attached leaves of pumpkin (Cucurbita pepo L.) were treated in high or moderate light at room temperature or a 1°C. The symptoms of photoinhibition appearing during light treatments at room temperature could be attributed to a decrease in the primary activity of PSII. However, when the light treatment was given at 1°C, the quantum yield of photosynthetic oxygen evolution decreased much more than would be expected from the decrease in the ratio of variable to maximum fluorescence at 77°K. Also, light treatment at 1°C lowered the chloroplast wholechain electron transfer capacity much more than it affected PSII electron transport (H₂O to paraphenylbenzoquinone). Light treatments at both room temperature and 1°C led to an increase in B_max, which indicates an increase in the proportion of PSII_β centers. PSI was not affected by the light treatments, and the treatments in the dark at 1°C caused only minor changes in the measured properties of the leaves. We conclude that high light always inhibits the primary activity of PSII, but at low temperature there is greater inhibition of electron transfer from primary electron accepting plastoquinone of PSII to the plastoquinone pool, which leads to a drastic decrease in the quantum yield of oxygen evolution in the chilling-sensitive pumpkin.