Thylakoid Pigment-protein Complexes Research Articles

High light-induced changes in whole-cell proteomic profile and its correlation with the organization of thylakoid super-complex in cyclic electron transport mutants of Chlamydomonas reinhardtii.

Light and nutrients are essential components of photosynthesis. Activating the signaling cascades is critical in starting adaptive processes in response to high light. In this study, we have used wild-type (WT), cyclic electron transport (CET) mutants like Proton Gradient Regulation (PGR) (PGRL1), and PGR5 to elucidate the actual role in regulation and assembly of photosynthetic pigment-protein complexes under high light. Here, we have correlated the biophysical, biochemical, and proteomic approaches to understand the targeted proteins and the organization of thylakoid pigment-protein complexes in the photoacclimation. The proteomic analysis showed that 320 proteins were significantly affected under high light compared to the control and are mainly involved in the photosynthetic electron transport chain, protein synthesis, metabolic process, glycolysis, and proteins involved in cytoskeleton assembly. Additionally, we observed that the cytochrome (Cyt) b6 expression is increased in the pgr5 mutant to regulate proton motive force and ATPase across the thylakoid membrane. The increased Cyt b6 function in pgr5 could be due to the compromised function of chloroplast (cp) ATP synthase subunits for energy generation and photoprotection under high light. Moreover, our proteome data show that the photosystem subunit II (PSBS) protein isoforms (PSBS1 and PSBS2) expressed more than the Light-Harvesting Complex Stress-Related (LHCSR) protein in pgr5 compared to WT and pgrl1 under high light. The immunoblot data shows the photosystem II proteins D1 and D2 accumulated more in pgrl1 and pgr5 than WT under high light. In high light, CP43 and CP47 showed a reduced amount in pgr5 under high light due to changes in chlorophyll and carotenoid content around the PSII protein, which coordinates as a cofactor for efficient energy transfer from the light-harvesting antenna to the photosystem core. BN-PAGE and circular dichroism studies indicate changes in macromolecular assembly and thylakoid super-complexes destacking in pgrl1 and pgr5 due to changes in the pigment-protein complexes under high light. Based on this study, we emphasize that this is an excellent aid in understanding the role of CET mutants in thylakoid protein abundances and super-complex organization under high light.

Differential response of photosynthetic apparatus towards alkaline pH treatment in NIES-39 and PCC 7345 strains of Arthrospira platensis.

Alkaline stress is one of the severe abiotic stresses, which is not well studied so far, especially among cyanobacteria. To affirm the characteristics of alkaline stress and the subsequent adaptive responses in Arthrospira platensis NIES-39 and Arthrospira platensis PCC 7345, photosynthetic pigments, spectral properties of thylakoids, PSII and PSI activities, and pigment-protein profiles of thylakoids under different pH regimes were examined. The accessory pigments showed a pH-mediated sensitivity. The pigment-protein complexes of thylakoids are also affected, resulting in the altered fluorescence emission profile. At pH 11, a possible shift of the PBsome antenna complex from PSII to PSI is observed. PSII reaction center is found to be more susceptible to alkaline stress in comparison to the PSI. In Arthrospira platensis NIES-39 at pH 11, a drop of 68% in the oxygen evolution with a significant increase of PSI activity by 114% is recorded within 24 h of pH treatment. Alterations in the cellular ultrastructure of Arthrospira platensis NIES-39 at pH 11 were observed, along with the increased number of plastoglobules attached with the thylakoid membranes. Arthrospira platensis NIES-39 is more adaptable to pH variation than Arthrospira platensis PCC 7345.

Small One-Helix Proteins Are Essential for Photosynthesis in Arabidopsis

The extended superfamily of chlorophyll a/b binding proteins comprises the Light-Harvesting Complex Proteins (LHCs), the Early Light-Induced Proteins (ELIPs) and the Photosystem II Subunit S (PSBS). The proteins of the ELIP family were proposed to function in photoprotection or assembly of thylakoid pigment-protein complexes and are further divided into subgroups with one to three transmembrane helices. Two small One-Helix Proteins (OHPs) are expressed constitutively in green plant tissues and their levels increase in response to light stress. In this study, we show that OHP1 and OHP2 are highly conserved in photosynthetic eukaryotes, but have probably evolved independently and have distinct functions in Arabidopsis. Mutations in OHP1 or OHP2 caused severe growth deficits, reduced pigmentation and disturbed thylakoid architecture. Surprisingly, the expression of OHP2 was severely reduced in ohp1 T-DNA insertion mutants and vice versa. In both ohp1 and ohp2 mutants, the levels of numerous photosystem components were strongly reduced and photosynthetic electron transport was almost undetectable. Accordingly, ohp1 and ohp2 mutants were dependent on external organic carbon sources for growth and did not produce seeds. Interestingly, the induction of ELIP1 expression and Cu/Zn superoxide dismutase activity in low light conditions indicated that ohp1 mutants constantly suffer from photo-oxidative stress. Based on these data, we propose that OHP1 and OHP2 play an essential role in the assembly or stabilization of photosynthetic pigment-protein complexes, especially photosystem reaction centers, in the thylakoid membrane.

Changes induced by cadmium stress and iron deficiency in the composition and organization of thylakoid complexes in sugar beet (Beta vulgaris L.)

In intact plants, Cd-induced Fe deficiency is thought to play a role in the toxic effects of Cd on photosynthesis. To investigate the contribution of the Cd-induced Fe deficiency to Cd stress symptoms we studied the composition and organization changes of thylakoid pigment–protein complexes by two-dimensional Blue Native-SDS gel electrophoresis and mass spectrometry, in parallel to functional changes, using Beta vulgaris plants grown in hydroponics. Plants were treated by withdrawing Fe or with 10μM CdCl2 for 10 days. Both metal stresses caused a marked decline in leaf chlorophyll concentration and chloroplast Fe content, as well as a loss of photosystem I (PSI) and light harvesting complex II (LHCII) complexes. Furthermore, marked organizational changes of the photosynthetic apparatus were found, including a decrease in the ratio of the PSII mega-/supercomplexes and an increase in the monomeric form of the LHCII antennae, with the extent of these changes being similar under both stresses. This supports that Fe deficiency responses have a major role in the responses of plants under Cd stress. In Fe-deficient thylakoids the increase in the ratio of PSI supercomplexes and degrading PSII particles was more pronounced, and higher zeaxanthin content was found. Under Cd stress, a stronger inhibition of PSII activity and enhancement of thermal dissipation of the inactive PSII complexes were observed. The differences detected under the two metal stresses lead to the conclusion that both local Fe deficiency in chloroplasts and other direct or indirect inhibitory effects of Cd are behind the response mechanisms of plants grown under Cd stress.

Photoacclimation in Spathiphyllum

We studied photoacclimation in Spathiphyllum grown at an irradiance of 40 or 420 μmol/m 2 s (LL or HL, respectively). All parameters studied responded to acclimation. Leaves at LL, in contrast to HL, were thinner and oriented perpendicular to the incident light, had more chlorophyll per g f w, fewer stomata on the upper leaf surface and a reduced layer of mesophyll cells. Their chloroplasts at HL had wider grana with less thylakoids per granum, and better organized photosystems than at LL. PSI and PSII activities per mg chlorophyll ( V max), and PSI and PSII content (total activity per g f w), were lower at LL than at HL and so was the light requirement for saturation of the PSI or PSII partial photoreactions, suggesting that fewer photosystems with larger antenna size prevail at LL, but many more with smaller antenna size at HL. Analysis of chlorophyll distribution among the thylakoid pigment–protein complexes showed less antenna chlorophyll serving PSII (CPa + LHCP 1 + LHCP 3) than that serving PSI (CPIa + CPI + LHCP 2) at LL as compared to HL, and thus a lower PSII/PSI ratio at LL, in agreement with the general finding that LL plants, with larger PSII antenna size, have lower PSII/PSI ratio. The increase in PSI antenna size at LL was correlated with the increase in the distribution of chlorophyll in pigment–protein complexes serving PSI, and a very large chlorophyll/protein molar ratio in the isolated CPI complex. On the other hand, the PSII antenna chlorophyll (CPa + LHCP 1 + LHCP 3) on a g f w basis, and the chlorophyll a/b ratio remained more or less constant at LL or HL. This may reflect our finding that Spathiphyllum contains mainly the 27 kDa inner LHCII antenna protein, the size of which remains unaffected by photoacclimation. The increase in the distribution of chlorophyll in pigment–protein complexes serving PSII at HL, therefore, reflects the higher population of PSII at HL. Very high PSI activity was found at HL, which we attribute to the highly organized small in size PSI.

A comparison of photosynthetic apparatus of the detached leaves of the resurrection plant Boea hygrometrica with its non-tolerant relative Chirita heterotrichia in response to dehydration and rehydration

Comparisons of the water contents in detached leaves of Boea hygrometrica (Bunge) R. Br ., and other members of the family Gesneriaceae have shown that B. hygrometrica can tolerate an extraordinarily high degree of water loss and restore its life upon rehydration. We have investigated the effects of dehydration on photosynthetic function in B. hygrometrica in comparison with its non-resurrection relative Chirita heterotrichia Merr. Studies on the net photosynthetic rate, chlorophyll fluorescence emission and excitation spectra, and the kinetics of chlorophyll fluorescence induction revealed that the effects of dehydration on both species were similar insofar as photosystem II was more sensitive to dehydration than photosystem I. However, B. hygrometrica responded more rapidly than C. heterotrichia. Although photosynthetic activity could be restored in both species upon rehydration after 2 days of dehydration, only B. hygrometrica recovered when the dehydration period was extended to 8 days. In contrast to C. heterotrichia leaves, in which the levels of carotenoids decreased rapidly during desiccation, carotenoid content actually increased in B. hygrometrica leaves upon dehydration and returned to the normal level after rehydration. Non-denaturing electrophoresis revealed that the thylakoid pigment–protein complexes of B. hygrometrica were highly stable during desiccation and rehydration, whereas those of C. heterotrichia were totally destroyed after 8 days of desiccation.

Effects of NaCl Stress on the Structure, Pigment Complex Composition, and Photosynthetic Activity of Mangrove Bruguiera parviflora Chloroplasts

Exposure of two-month-old seedlings of Bruguiera parviflora to NaCl stress (0 to 400 mM) for 45 d under hydroponic culture caused notable disorganisation of the thylakoid structure of chloroplasts in NaCl-treated leaves as revealed from transmission electron microscopy. The absorption spectra of treated and control thylakoid samples were similar having a red peak at 680 nm and Soret peaks at 439 and 471 nm in the blue region of the spectrum. The spectra of treated samples differed from control samples by gradual decrease in absorbance of 100, 200, and 400 mM NaCl treated samples at 471 and 439 nm, which could be due to scattering of radiation in these samples. Thus, absorption characteristics of thylakoid membranes indicated no major alterations in the structural integrity of the photosynthetic membranes during salt stress in B. parviflora. Analysis of pigment protein complexes of thylakoids on non-denaturing gel showed that CP1 complex consisting of photosystem (PS) 1 reaction centre decreased marginally by 19% and the CP47 constituting the core antenna of PS2 declined significantly by 30% in 400 mM NaCl treated samples in respect to control. This decrease in structural core antenna might cause inefficient photon harvesting capacity. However, CP43 content did not alter. An increase in CP2/CP1 ratio from 3.2 in control to 4.0 in 400 mM NaCl treated samples indicated significant structural changes in the thylakoids of salt treated plants. Haem staining of thylakoids revealed significant losses in cytochrome (Cyt)f and Cyt b6 contents by NaCl stress. However, Cyt b559 content remained nearly constant in both control and NaCl treated samples. SDS-PAGE of thylakoid proteins showed that the intensity of many of Coomassie stained polypeptide bands ranging from 15–22 and 28–66 kDa regions decreased significantly in NaCl treated samples as compared to control. Electron transport activity of thylakoids, measured in terms of DCPIP photoreduction, was 22% lower in 400 mM NaCl treated plants than in the control ones. Hence, NaCl induces oxidative stress in chloroplasts causing structural alterations in thylakoids. These structural alterations might be responsible for declined efficiency of photosystems and reduced electron transport activity.

Photosynthetic energy conversion under extreme conditions—II: the significance of lipids under light limited growth in Antarctic sea ice diatoms

Low photosynthetic active radiation is a strong determinant in the development and growth of sea ice algae. The algae appear to have universal mechanisms to overcome light limitation. One important process, which is induced under light limitation, is the desaturation of chloroplast membrane lipids. In order to discover whether this process is universally valid in sea ice diatoms, we investigated three species coexisting in chemostats illuminated with 15 and 2 μmol photons m −2 s −1 at −1 °C. Growth under 2 μmol photons m −2 s −1 caused a 50% increase in monogalactosyldiacylglycerols (MGDG) thylakoid membrane related 20:5 n-3 fatty acids. This fatty acid supports the fluidity of the thylakoid membrane and therefore the velocity of electron flow, which is indicated by increasing rate constants for the electron transport between Q A (first stable electron acceptor) and bound Q B (second stable electron acceptor) (11.16±1.34 to 23.24±1.35 relative units). Two μmol photons m −2 s −1 furthermore resulted in higher amounts of non-lipid bilayer forming MGDG in relation to other bilayer forming lipids, especially digalactosydiacylglycerol (DGDG). The ratio of MGDG:DGDG increased from 3.4±0.3 to 5.7±0.3. The existence of bilayer thylakoid membranes with high proportions of non. bilayer forming lipids is only possible when sufficient thylakoid pigment–protein complexes are present. If more thylakoid pigment–protein complexes are present in membranes, as found under extreme light limitation, less bilayer forming lipids such as DGDG are required to stabilize the bilayer structure. Differences in protein contents between both light intensities were not found. Consequently pigment contents which nearly doubled under 2 μmol photons m −2 s −1 must be responsible in balancing the potential stability loss resulting from an increase in MGDG:DGDG ratio.

Leaf C40.4: a carotenoid-associated protein involved in the modulation of photosynthetic efficiency?

Comparative analysis by differential RNA display (DDRT-PCR) of the expression patterns of potato plants induced and non-induced to tuberize, led to the isolation of a cDNA clone, C40.4, that is strongly upregulated in the leaves of tuberizing plants. Leaf expression of this transcript was shown to be light-dependent, with increased levels of mRNA and protein being detected during the light hours. Sequence analysis revealed near complete identity to potato CDSP34, a thylakoid protein induced by drought stress, and strong homology to the carotenoid-associated proteins fibrillin, ChrB and PAP from pepper fruit, and CHRC from cucumber flowers. By using an antibody against pepper fibrillin, we were able to demonstrate a chloroplastic location of the C40.4 protein in association with the thylakoid membranes. Two-dimensional analysis of thylakoid pigment-protein complexes showed a specific association of the protein with the photosystem II (PSII) multi-subunit complex. Antisense plants with reduced levels of accumulation of C40.4 showed a stunted growth and decreased tuber yield and exhibited reduced values of non-photochemical quenching of chlorophyll a fluorescence. Altogether, these results indicate a preferential association of the C40.4 protein with the light harvesting complex of photosystem II (LHCII) antenna pigment complexes, and suggest a functional role of this protein in photosynthesis, by modulating photosynthetic efficiency and dissipation of excess absorbed light energy within the antenna complex.

Photosynthetic apparatus in primary leaves of barley seedlings grown under blue or red light of very low photon flux densities

Chlorophyll (Chl) a and Chl b contents, rate of CO2 gas exchange, quenching coefficients of chlorophyll fluorescence, and endogenous phytohormones have been studied in primary leaves of barley seedlings cultivated under blue (BL) or red (RL) light. Photon flux densities (PFD) were between 0.3 and 12 μmol m-2 s-1. Plants grown at PFD of 0.3 μmol m-2 s-1 demonstrated in BL tenfold and in RL threefold decreased Chl content compared to plants grown at 12 μmol m-2 s-1. Chl a/b ratio increased from 2.3–2.5 to 4.4–4.5 in BL, not in RL, following the decrease in PFD at plant cultivation from 12 to 0.3 μmol m-2 s-1. Plants cultivated at weak BL demonstrated severalfold decreased rate of photosynthetic CO2 uptake, whereas decrease in PFD of RL from 12 to 0.3 μmol m-2 s-1 caused only 20% de cline in the rate of photosynthesis. Decrease in PFD during a plant cultivation reduced the maximum quantum yield of photosynthesis in BL, not in RL leaves. Light response curves of non-photochemical and photochemical quenching of chlorophyll fluorescence calculated on the basis of absorbed quanta were not affected by PFD of RL during plant cultivation. On the contrary, both non-photochemical quenching and accumulation of QA-, reduced primary acceptor of Photosystem II, occurred at lower amounts of absorbed quanta in leaves of BL plants grown at 0.3 than at 12 μmol m-2 s-1. Two photoregulatory reactions were suggested to exert the light control of the development of photosynthetic apparatus in the range of low PFDs. The photoregulatory reaction saturating by very low PFDs of RL was supposed to be mediated by phytochrome. Phytochrome was proposed to enhance (as related to other pigment-protein complexes of thylakoids) the accu mulation of chlorophyll- b-binding light-harvesting complex of Photosystem II (LHC II). It acts independently of the pigment mediating the second photoregulatory reaction, as evidenced by the results of experiments with plant growth under mixed blue plus red light. The contents of cytokinins and indole-3-acetic acid in a leaf were not significantly affected by either light quality and PFD thus indicating those phytohormones not to be involved into photoregulatory processes.

The coleoptile chloroplast: Distinct distribution of xanthophyll cycle pigments, and enrichment in Photosystem II

Recent studies have shown that coleoptile chloroplasts operate the xanthophyll cycle, and that their zeaxanthin concentration co-varies with their sensitivity to blue light. The present study characterized the distribution of photosynthetic pigments in thylakoid pigment–protein complexes from dark-adapted and light-treated coleoptile and mesophyll chloroplasts, the low temperature fluorescence emission spectra, and the rates of PS I and PS II electron transport in both types of chloroplasts from 5-day-old corn seedlings. Pigments were extracted from isolated PS I holocomplex, LHC IIb trimeric and LHC II monomeric complexes and analyzed by HPLC. Chlorophyll distribution in coleoptile thylakoids showed 31% of the total collected Chl in PS I and 65% in the light harvesting complexes of PS II. In mesophyll thylakoids, the values were 44% and 54%, respectively. Mesophyll and coleoptile PS I holocomplexes differed in their Chl t a/Chl t b ratios (8.1 and 6.1, respectively) and β-carotene content. In contrast, mesophyll and coleoptile LHC IIb trimers and LHC II monomers had similar Chl t a/Chl t b ratios and β-carotene content. The three analyzed pigment–protein complexes from dark-adapted coleoptile chloroplasts contained zeaxanthin, whereas there was no detectable zeaxanthin in the complexes from dark-adapted mesophyll chloroplasts. In both chloroplast types, zeaxanthin and antheraxanthin increased markedly in the three pigment–protein complexes upon illumination, while violaxanthin decreased. In mesophyll thylakoids, zeaxanthin distribution as a percentage of the xanthophyll cycle pool was: LHC II monomers > LHC IIb trimers > PS I holocomplex, and in coleoptile thylakoids, it was: LHC IIb trimers > LHC II monomers = PS I holocomplex. Low temperature (77 K) fluorescence emission spectra showed that the 686 nm emission of coleoptile chloroplasts was approximately 50% larger than that of mesophyll chloroplasts when normalized at 734 nm. The pigment and fluorescence analysis data suggest that there is relatively more PS II per PS I and more LHC I per CC I in coleoptile chloroplasts than in mesophyll chloroplasts. Measurements of t in vitro uncoupled photosynthetic electron transport showed approximately 60% higher rates of electron flow through PS II in coleoptile chloroplasts than in mesophyll chloroplasts. Electron transport rates through PS I were similar in both chloroplast types. Thus, when compared to mesophyll chloroplasts, coleoptile chloroplasts have a distinct PS I pigment composition, a distinct chlorophyll distribution between PS I and PS II, a distinct zeaxanthin percentage distribution among thylakoid pigment–protein complexes, a higher PS II-related fluorescence emission, and higher PS II electron transport capacity. These characteristics may be associated with a sensory transducing role of coleoptile chloroplasts.

Heat-Induced Alterations in Thylakoid Membrane Protein Composition in Barley

Biochemical and spectroscopic studies on the effects of high temperatures (45-47� C) over a 1 h period on the protein composition, fluorescence and photochemical activities of the barley thylakoid membrane were made. Photosystem II (PS II) activity decreased as expected, and photosystem I (PS I) activity also unexpectedly decreased. Our data support previous conclusions that the decrease in PS I activity is largely due to inactivation (or loss) of a component between the two photosystems. A two-dimensional electrophoretic system permitted first the separation of the thylakoid pigment-protein complexes of unstressed and stressed plants, followed by a determination of their subunit composition. The changes in the protein composition of each pigment-protein complex in response to elevated temperatures were monitored. Heat changed the quaternary structure of PS II and resulted in removal of the oxygen-evolving enhancer proteins from the thylakoid, but did essentially no damage to the PS I complex. The PS II core complex dissociated from a dimeric form to a monomeric one, and the major LHC II component (LHC IIb) changed from a trimeric to a monomeric form. The pigments that are lost from thylakoids during heat stress are mainly removed from the PS II pigment-proteins.

Carotenoid distribution and deepoxidation in thylakoid pigment-protein complexes from cotton leaves and bundle-sheath cells of maize

In response to excess light, the xanthophyll violaxanthin (V) is deepoxidized to zeaxanthin (Z) via antheraxanthin (A) and the degree of this deepoxidation is strongly correlated with dissipation of excess energy and photoprotection in PS II. However, little is known about the site of V deepoxidation and the localization of Z within the thylakoid membranes. To gain insight into this problem, thylakoids were isolated from cotton leaves and bundle-sheath strands of maize, the pigment protein-complexes separated on Deriphat gels, electroeluted, and the pigments analyzed by HPLC. In cotton thylakoids, 30% of the xanthophyll cycle pigments were associated with the PS I holocomplex, including the PS I light-harvesting complexes and PS I core complex proteins (CC I), and about 50% with the PS II light-harvesting complexes (LHC II). The Chl was evenly distributed between PS I and PS II. Less than 2% of the neoxanthin, about 18% of the lutein, and as much as 76% of the β-carotene of the thylakoids were associated with PS I. Exposure of pre-darkened cotton leaves to a high photon flux density for 20 min prior to thylakoid isolation caused about one-half of the V to be converted to Z. The distribution of Z among the pigment-protein complexes was found to be similar to that of V. The distribution of the other carotenoids was unaffected by the light treatment. Similarly, in field-grown maize leaves and in the bundle-sheath strands isolated from them, about 40% of the V present at dawn had been converted to Z at solar noon. Light treatment of isolated bundle-sheath strands which initially contained little Z caused a similar degree of conversion of V to Z. As in cotton thylakoids, about 30% the V+A+Z pool in bundle-sheath thylakoids from maize was associated with the PS I holocomplex and the CC I bands and 46% with the LHC II bands, regardless of the extent of deepoxidation. These results demonstrate that Z is present in PS I as well as in PS II and that deepoxidation evidently takes place within the pigment-protein complexes of both photosystems.

Correlation between the “Low”-salt-induced increase in the F730/F685 fluorescence emission ratio at 77 K in isolated chloroplasts, and the organization of chlorophyll in photosystem I pigment-protein complexes of thylakoids

Isolated pea or spinach chloroplasts suspended in “high”-salt phosphate buffer exhibit a low F730/F685 fluorescence emission ratio at 77 K; in contrast, removal of cations by incubation in “low”-salt Tricine buffer induces a drastic increase in the F730/F685 ratio. Parallel to the F730/F685 ratio increase, a gradual organization of chlorophyll (Chl) in the pigment-protein complexes of the Photosystem I, chlorophyll-protein complex Ia, and light-harvesting complex I (LHC-I), is observed. The kinetics of the two processes are closely correlated, all changes being completed within 5–10 min from Tricine addition. On the other hand, the inability of low-salt Tricine to induce any changes in the F730/F685 ratio in bean plastids, isolated and suspended in high-salt phosphate buffer, correlates with the lack of extensive changes in the organization of the Photosystem I complexes, and more specifically of LHC-I. The latter is attributed to the higher stability of complexes in bean, arising from stronger association of thylakoids in grana stacks in this species; this is probably due to higher levels of residual divalent cations present in the isolated thylakoids of bean compared to pea (or spinach). The results suggest that the F730/F685 ratio changes, observed in chloroplasts by manipulation of their ionic environment, reflect modulation of Chl organization within the pigment-protein complexes of the photosynthetic units.

Uniconazole‐induced thermotolerance in wheat seedlings is mediated by transpirational cooling

Seed treatment of thermotolerant and sensitive cultivars of wheat (Triticum aestivum L. cv. Frederick and Glenlea, respectively) with uniconazole reduced shoot and increased root fresh and dry weights. When subjected to 48°C for 6 h, treated seedlings had lower leaf‐temperatures and overall higher rates of evapotranspiration. Percent survival 11 days after high temperature incubation in untreated seedlings was 53% in Frederick and 30% in Glenlea, whereas in uniconazole‐treated seedlings it was 94% and 80%, respectively. Transpiration resistance markedly increased in the control cultivars after 4 h of high‐temperature incubation, but remained relatively unchanged in the treated cultivars. The increased root to shoot ratio exhibited in treated plants may have alleviated guard cell stress, even under high evaporative demand. Chlorophyll fluorescence measurements on leaves of heat‐stressed seedlings indicated that increased photosynthetic metabolism in treated seedlings was correlated with lower leaf temperatures. High‐temperature stress resulted in a dramatic decline in pigments and proteins in thylakoid extracts of control seedlings but not in extracts from treated seedlings. Integrity of thylakoid pigment protein complexes, as illustrated by reduced relative amounts of free pigment, was maintained after exposure to high temperatures in treated seedlings. Furthermore, the treated Glenlea seedlings displayed a reduction in the monomeric form of Sight‐harvesting chlorophyll protein II (LHCP II) compared to control and Frederick seedlings. The heat‐tolerant Frederick cultivar showed a greater protective effect from uniconazole treatment than the sensitive Glenlea cultivar. Uniconazole treatment did not affect heat‐shock protein (HSP) synthesis in mesocotyl tissue.

Modifications to Thylakoid Composition during Development of Maize Leaves at Low Growth Temperatures

The effects of reductions in growth temperature on the development of thylakoids of maize (Zea mays var LG11) leaves are examined. Thylakoids isolated from mesophyll cells of leaves grown at 17 degrees and 14 degrees C, compared with 25 degrees C, exhibited a decreased accumulation of many polypeptides, which was accompanied by a loss of activity of photosystems (PS) I and II. Probing the polypeptide profiles with a range of antibodies specific for thylakoid proteins demonstrated that a number of polypeptides encoded by the chloroplast genome failed to accumulate at low temperatures. Although thylakoid protein synthesis was reduced severely at 14 degrees C compared with 25 degrees C, major synthesis of both chloroplast and nuclear encoded polypeptides was detected. It is suggested that the lack of accumulation of some thylakoid proteins at low temperatures may be due to an inability to stabilize the proteins in the membranes. A number of thylakoid polypeptides were found to appear as the growth temperature was decreased. Analyses of pigments and polypeptides demonstrated that decreases in the photosystem reaction center core complexes occur relative to the light harvesting complex associated with PS II at reduced growth temperatures. Differential effects on the development of PSI and PSII were also observed, with PSII activity being preferentially reduced. Reductions in PSII content and activity occurred in parallel with decreases in the quantum yield and light-saturated rate of CO(2) assimilation. Fractionation of thylakoid pigment-protein complexes showed that the ratio of monomeric:oligomeric form of the light harvesting complex associated with PSII increased at low growth temperature, which is consistent with a chill-induced modification of thylakoid organization. Many, but not all, of the characteristic changes in thylakoid protein metabolism, which were observed when leaves were grown at low temperatures in controlled environments, were identified in leaves of a field maize crop during the early growing season when low temperatures were experienced by the crop. Chill-induced perturbations of thylakoid development can occur in the field in temperate regions and may have implications for the photosynthetic productivity of the crop.

Galactolipids of Thylakoid Pigment Protein Complexes Separated Electrophoretically from Thylakoids of Dunaliella salina Labeled with Radioactive Fatty Acids

Experiments have been conducted to determine whether mono- and digalactosyldiacylglycerols (MGDG and DGDG) labeled by short term exposure of the green alga Dunaliella salina to exogenous radioactive lauric, myristic, or palmitic acids are preferentially recovered in one or more thylakoid protein complexes separated by electrophoresis on nondenaturing gels. A selective retention of certain lipid classes and molecular species by the major electrophoretically separable fractions, oligomeric form (LHCP-1) and monomeric form (LHCP-3) of hight-harvesting chlorophyll a/b protein complex, and major photosystem I protein complex (CPla), and nonprotein-bound lipids, was indicated. After 5 min of labeling with (3)H-palmitate the specific radioactivities of MGDG and DGDG molecular species recovered from LHCP-3 and CPla were slightly higher and lower, respectively, than specific radioactivities of equivalent molecular species from LHCP-1 and nonprotein-bound lipid. However, the differences were smaller than those which have been observed between whole thylakoids and the more highly labeled chloroplast envelopes from briefly labeled cells.

The distribution of α-carotene in the photosynthetic pigment-protein complexes of higher plants

α-Carotene (β, ϵ-carotene) is a minor component of the chloroplast carotenoids in many plant species, but its location in the thylakoid pigment-protein complexes has never been established. It is now shown that in the dwarf French Bean ( Phaseolus vulgaris) the distribution of α-carotene in the complexes is strongly correlated with that of β-carotene, i.e. α-carotene is associated primarily with the photosystem I (PSI) and photosystem II (PSII) reaction centres, with only small quantities in the light-harvesting complexes.

Supramolecular structure of chlorophyll-protein complexes in relation to the chlorophyll a fluorescence of chloroplasts at room or liquid nitrogen temperature

To investigate further the possibility that changes in the organization of the thylakoid pigment-protein complexes are monitored by the chlorophyll a fluorescence yield changes, or by changes in the F 685 F 730 ratio in the 77 °K fluorescence, as earlier proposed ( A. Castorinis, G. Akoyunoglou, and J. H. Argyroudi-Akoyunoglou (1982) Photobiochem. Photobiophys. 4, 283–291 ; J. H. Argyroudi-Akoyunoglou, A. Castorinis, and G. Akoyunoglou (1982) Photobiochem. Photobiophys. 4, 201–210 ), the effect of pH, Cd 2+ or Zn 2+ addition, and trypsinization on all parameters was studied. We found that (a) the pH of the medium affects the ratio of oligomeric to monomeric structures of the complexes (monomers predominate at low pH), as well as the ratio F 685 F 730 of thylakoids or of the isolated CPIa complex (the F 685 F 730 ratio is high at low pH, and low at high). (b) Zn 2+ or Cd 2+ addition to thylakoids, suspended in Tricine ( N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine), has no effect on the chlorophyll a fluorescence yield nor on the F 685 F 730 ratio at 77 °K; similarly, no effect can be noticed on the F 685 F 730 ratio at 77 °K of the isolated CPIa complex. These cations, contrary to Mg 2+, do not affect the oligomer to monomer dissociation, but they instead reverse the cation effect and enhance the oligomeric structures. (c) Trypsinized thylakoids in Tricine do not show the Mg 2+ effect on the chlorophyll a fluorescence yield, have a reduced F 730 F 685 ratio at 77 °K, and are deficient in the CPIa complex, which is mainly responsible for the F 685 F 730 ratio changes in the presence of cations. The results support the proposal that cation- or pH-induced changes in the chlorophyll a fluorescence yield at room temperature, or in the F 685 F 730 ratio at 77 °K, are irrelevant to the excitation energy distribution between the two photosystems. They instead seem to reflect changes in the organization of the supramolecular structure of the pigment-protein complexes in the photosystem I and photosystem II units.

Separation of thylakoid pigment—protein complexes by SDS—sucrose density gradient centrifugation

Separation of thylakoid pigment—protein complexes by SDS—sucrose density gradient centrifugation