Chloroplasts Of Green Plants Research Articles

Proteomes reveal the lipid metabolic network in the complex plastid of Phaeodactylum tricornutum.

Phaeodactylum tricornutum plastid is surrounded by four membranes, and its protein composition and function remain mysterious. In this study, the P. tricornutum plastid-enriched fraction was obtained and 2850 proteins were identified, including 92 plastid-encoded proteins, through label-free quantitative proteomic technology. Among them, 839 nuclear-encoded proteins were further determined to be plastidial proteins based on the BLAST alignments within Plant Proteome DataBase and subcellular localization prediction, in spite of the strong contamination by mitochondria-encoded proteins and putative plasma membrane proteins. According to our proteomic data, we reconstructed the metabolic pathways and highlighted the hybrid nature of this diatom plastid. Triacylglycerol (TAG) hydrolysis and glycolysis, as well as photosynthesis, glycan metabolism, and tocopherol and triterpene biosynthesis, occur in the plastid. In addition, the synthesis of long-chain acyl-CoAs, elongation, and desaturation of fatty acids (FAs), and synthesis of lipids including TAG are confined in the four-layered-membrane plastid based on the proteomic and GFP-fusion localization data. The whole process of generation of docosahexaenoic acid (22:6) from palmitic acid (16:0), via elongation and desaturation of FAs, occurs in the chloroplast endoplasmic reticulum membrane, the outermost membrane of the plastid. Desaturation that generates 16:4 from 16:0 occurs in the plastid stroma and outer envelope membrane. Quantitative analysis of glycerolipids between whole cells and isolated plastids shows similar composition, and the FA profile of TAG was not different. This study shows that the diatom plastid combines functions usually separated in photosynthetic eukaryotes, and differs from green alga and plant chloroplasts by undertaking the whole process of lipid biosynthesis.

Thylakoids: A Novel Food-Derived Supplement for Obesity - A Mini-Review.

Nowadays, overweight and obesity are major epidemic health problems that can bring about some other health issues such as cardiovascular disease which is the first cause of mortality worldwide. Thylakoids are disc-like membranes responsible for photosynthetic light reactions in chloroplasts of green plants. Although only a few animal and human studies have been conducted regarding the impact of thylakoids on overweight- and obesity-related factors, all of them have resulted in positive outcomes. These outcomes are as follows: increment of satiety response; suppression of hunger sensations, particularly hedonic hunger; reduction of body weight and fat; promotion of glucose homeostasis; decrease in serum lipids; attenuation of oxidative stress and inflammation; and modulation of gut microbiota, notably by increasing some helpful bacteria such as Lactobacillus reuteri. It seems that some of these useful effects are related to retarded absorption of dietary fat and carbohydrate caused by thylakoids. There is still a need for more well-designed studies.

Structure, mechanism, and regulation of the chloroplast ATP synthase

INTRODUCTION Green plant chloroplasts convert light into chemical energy, and adenosine triphosphate (ATP) generated by photosynthesis is the prime source of biologically useful energy on the planet. Plants produce ATP by the chloroplast F 1 F o ATP synthase (cF 1 F o ), a macromolecular machine par excellence, driven by the electrochemical proton gradient across the photosynthetic membrane. It consists of 26 protein subunits, 17 of them wholly or partly membrane-embedded. ATP synthesis in the hydrophilic α 3 β 3 head (cF 1 ) is powered by the cF o rotary motor in the membrane. cF o contains a rotor ring of 14 c subunits, each with a conserved protonatable glutamate. Subunit a conducts the protons to and from the c-ring protonation sites. The central stalk of subunits γ and e transmits the torque from the F o motor to the catalytic cF 1 head, resulting in the synthesis of three ATP per revolution. The peripheral stalk subunits b, b′, and δ act as a stator to prevent unproductive rotation of cF 1 with cF o . All rotary ATP synthases are, in principle, fully reversible. To prevent wasteful ATP hydrolysis, cF 1 F o has a redox switch that inhibits adenosine triphosphatase (ATPase) activity in the dark. RATIONALE Understanding the molecular mechanisms of this elaborate nanomachine requires detailed structures of the whole complex, ideally at atomic resolution. Because of the dynamic nature of this membrane protein complex, crystallization has been difficult and no high-resolution structure of an entire, functional ATP synthase is available. We reconstituted cF 1 F o from spinach chloroplasts into lipid nanodiscs and determined its structure by cryo–electron microscopy (cryo-EM). Cryo-EM is the ideal technique for this study because it can deliver high-resolution structures of large, dynamic macromolecular assemblies that adopt a mixture of conformational states. RESULTS We present the cryo-EM structure of the intact cF 1 F o ATP synthase in lipid nanodiscs at a resolution of 2.9 A (cF 1 ) to 3.4 A (cF o ). In the cF 1 ATPase head, we observe nucleotides with their coordinating Mg ions and water molecules, allowing assignment to the three well-characterized functional states involved in rotary ATP synthesis. Subunit δ on top of the ATPase head binds to all three α subunits, ensuring that only one peripheral stalk can attach. The loosely entwined, long α helices of the peripheral stalk subunits b and b′ clamp the integral membrane subunit a in its position next to the c-ring rotor, thus connecting cF 1 to cF o . Subunit γ has an L-shaped double hairpin with a redox sensor that can form a disulfide bond and a chock that blocks rotation to avoid wasteful ATP hydrolysis at night. Protons are translocated through access routes in subunit a in all rotary ATPases. We observe a hydrophilic channel on the lumenal surface that connects to the glutamate residues on the c-ring rotor that carry protons for an almost full rotation before releasing them into the stroma through another hydrophilic channel. A strictly conserved arginine separates the access and exit channels, preventing leakage of protons through the membrane. CONCLUSION We observe three cF 1 F o conformations, each with the central rotor stalled in a different position. Ring rotation is unexpectedly divided into three unequal steps. The peripheral stalk may thus act like an elastic spring, evening out the different energy contributions of each step. The features of ATP synthase nanomachines are remarkably similar in chloroplasts and mitochondria, considering their evolutionary distance of a billion years or more.

Two paths diverged in the stroma: targeting to dual SEC translocase systems in chloroplasts.

Chloroplasts inherited systems and strategies for protein targeting, translocation, and integration from their cyanobacterial ancestor. Unlike cyanobacteria however, chloroplasts in green algae and plants contain two distinct SEC translocase/integrase systems: the SEC1 system in the thylakoid membrane and the SEC2 system in the inner envelope membrane. This review summarizes the mode of action of SEC translocases, identification of components of the SEC2 system, evolutionary history of SCY and SECA genes, and previous work on the co- and post-translational targeting of lumenal and thylakoid membrane proteins to the SEC1 system. Recent work identifying substrates for the SEC2 system and potential features that may contribute to inner envelope targeting are also discussed.

Thioredoxin regulates G6PDH activity by changing redox states of OpcA in the nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120.

Glucose 6-phosphate dehydrogenase (G6PDH) catalyzes the first reaction in the oxidative pentose phosphate pathway. In green plant chloroplasts, G6PDH is a unique redox-regulated enzyme, since it is inactivated under the reducing conditions. This regulation is accomplished using a redox-active cysteine pair, which is conserved in plant G6PDH. The inactivation of this enzyme under conditions of light must be beneficial to prevent release of CO2 from the photosynthetic carbon fixation cycle. In the filamentous, heterocyst-forming, nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120 (Anabaena 7120), G6PDH plays a pivotal role in providing reducing power for nitrogenase, and its activity is also reported to be suppressed by reduction, though Anabaena G6PDH does not conserve the critical cysteines for regulation. Based on the thorough analyses of the redox regulation mechanisms of G6PDH from Anabaena 7120 and its activator protein OpcA, we found that m-type thioredoxin regulates G6PDH activity by changing the redox states of OpcA. Mass spectrometric analysis and mutagenesis studies indicate that Cys393 and Cys399 of OpcA are responsible for the redox regulation property of this protein. Moreover, in vivo analyses of the redox states of OpcA showed that more than half of the OpcA is present as an oxidized form, even under conditions of light, when cells are cultured under the nitrogen-fixing conditions. This redox regulation of OpcA might be necessary to provide reducing power for nitrogenase by G6PDH in heterocysts even during the day.

Chloroplast genome expansion by intron multiplication in the basal psychrophilic euglenoid Eutreptiella pomquetensis.

BackgroundOver the last few years multiple studies have been published showing a great diversity in size of chloroplast genomes (cpGenomes), and in the arrangement of gene clusters, in the Euglenales. However, while these genomes provided important insights into the evolution of cpGenomes across the Euglenales and within their genera, only two genomes were analyzed in regard to genomic variability between and within Euglenales and Eutreptiales. To better understand the dynamics of chloroplast genome evolution in early evolving Eutreptiales, this study focused on the cpGenome of Eutreptiella pomquetensis, and the spread and peculiarities of introns.MethodsThe Etl. pomquetensis cpGenome was sequenced, annotated and afterwards examined in structure, size, gene order and intron content. These features were compared with other euglenoid cpGenomes as well as those of prasinophyte green algae, including Pyramimonas parkeae.Results and DiscussionWith about 130,561 bp the chloroplast genome of Etl. pomquetensis, a basal taxon in the phototrophic euglenoids, was considerably larger than the two other Eutreptiales cpGenomes sequenced so far. Although the detected quadripartite structure resembled most green algae and plant chloroplast genomes, the gene content of the single copy regions in Etl. pomquetensis was completely different from those observed in green algae and plants. The gene composition of Etl. pomquetensis was extensively changed and turned out to be almost identical to other Eutreptiales and Euglenales, and not to P. parkeae. Furthermore, the cpGenome of Etl. pomquetensis was unexpectedly permeated by a high number of introns, which led to a substantially larger genome. The 51 identified introns of Etl. pomquetensis showed two major unique features: (i) more than half of the introns displayed a high level of pairwise identities; (ii) no group III introns could be identified in the protein coding genes. These findings support the hypothesis that group III introns are degenerated group II introns and evolved later.

The chloroplast-localized small heat shock protein Hsp21 associates with the thylakoid membranes in heat-stressed plants.

The small heat shock protein (sHsp) chaperones are crucial for cell survival and can prevent aggregation of client proteins that partially unfold under destabilizing conditions. Most investigations on the chaperone activity of sHsps are based on a limited set of thermosensitive model substrate client proteins since the endogenous targets are often not known. There is a high diversity among sHsps with a single conserved β‐sandwich fold domain defining the family, the α‐crystallin domain, whereas the N‐terminal and C‐terminal regions are highly variable in length and sequence among various sHsps and conserved only within orthologues. The endogenous targets are probably also varying among various sHsps, cellular compartments, cell type and organism. Here we have investigated Hsp21, a non‐metazoan sHsp expressed in the chloroplasts in green plants which experience huge environmental fluctuations not least in temperature. We describe how Hsp21 can also interact with the chloroplast thylakoid membranes, both when isolated thylakoid membranes are incubated with Hsp21 protein and when plants are heat‐stressed. The amount of Hsp21 associated with the thylakoid membranes was precisely determined by quantitative mass spectrometry after metabolic 15N‐isotope labeling of either recombinantly expressed and purified Hsp21 protein or intact Arabidopsis thaliana plants. We found that Hsp21 is among few proteins that become associated with the thylakoid membranes in heat‐stressed plants, and that approximately two thirds of the pool of chloroplast Hsp21 is affected. We conclude that for a complete picture of the role of sHsps in plant stress resistance also their association with the membranes should be considered.

Why we need to know the structure of phosphorylated chloroplast light-harvesting complex II.

In oxygenic photosynthesis there are two 'light states'-adaptations of the photosynthetic apparatus to spectral composition that otherwise favours either photosystem I or photosystem II. In chloroplasts of green plants the transition to light state 2 depends on phosphorylation of apoproteins of a membrane-intrinsic antenna, the chlorophyll-a/b-binding, light-harvesting complex II (LHC II), and on the resulting redistribution of absorbed excitation energy from photosystem II to photosystem I. The transition to light state 1 reverses these events and requires a phospho-LHC II phosphatase. Current structures of LHC II reveal little about possible steric effects of phosphorylation. The surface-exposed N-terminal domain of an LHC II polypeptide contains its phosphorylation site and is disordered in its unphosphorylated form. A molecular recognition hypothesis proposes that state transitions are a consequence of movement of LHC II between binding sites on photosystems I and II. In state 1, LHC II forms part of the antenna of photosystem II. In state 2, a unique but as yet unidentified 3-D structure of phospho-LHC II may attach it instead to photosystem I. One possibility is that the LHC II N-terminus becomes ordered upon phosphorylation, adopting a local alpha-helical secondary structure that initiates changes in LHC II tertiary and quaternary structure that sever contact with photosystem II while securing contact with photosystem I. In order to understand redistribution of absorbed excitation energy in photosynthesis we need to know the structure of LHC II in its phosphorylated form, and in its complex with photosystem I.

Effect of Cross-linking on the Performance and Stability of Photocatalytic Photosystem I Films

Extracted from the chloroplasts of green plants, Photosystem I (PSI) is a multi-subunit protein complex that can be deposited as a thin film onto electrodes and incorporated into biohybrid photovoltaic devices to convert sunlight into usable energy. Previously, our group characterized persistent photocurrents over nine months using PSI multilayer films in an aqueous solution of redox mediators. However, the loss of periphery PSI proteins upon exposure to mediator solutions is known to affect the photocatalytic performance of the overall film. The cross-linker 2-iminothiolane (2IT) is capable of connecting protein molecules with covalent bonds, and is herein used to stabilize PSI multilayer films on gold substrates against significant desorption/degradation. Results showed that the thickness retainment of the cross-linked PSI films was superior to that of non-cross-linked films. PSI films modified by 2IT maintained production of photocurrent with long-term exposure to aqueous solutions in contrast to non-cross-linked control films.

Evolution of the Cp-Actin-based Motility System of Chloroplasts in Green Plants.

During the course of green plant evolution, numerous light responses have arisen that optimize their growth under fluctuating light conditions. The blue light receptor phototropin mediates several photomovement responses at the tissue, cellular and organelle levels. Chloroplast photorelocation movement is one such photomovement response, and is found not only in most green plants, but also in some red algae and photosynthetic stramenopiles. In general, chloroplasts move toward weak light to maximally capture photosynthetically active radiation (the chloroplast accumulation response), and they move away from strong light to avoid photodamage (the avoidance response). In land plants, chloroplast movement is dependent on specialized actin filaments, chloroplast-actin filaments (cp-actin filaments). Through molecular genetic analysis using Arabidopsis thaliana, many molecular factors that regulate chloroplast photorelocation were identified. In this Perspective, we discuss the evolutionary history of the molecular mechanism for chloroplast photorelocation movement in green plants in view of cp-actin filaments.

Light-harvesting superstructures of green plant chloroplasts lacking photosystems.

The light-harvesting antenna of higher plant photosystem II (LHCII) is the major photosynthetic membrane component encoded by an entire family of homologous nuclear genes. On the contrary, the great majority of proteins of photosystems and electron transport components are encoded by the chloroplast genome. In this work, we succeeded in gradually inhibiting the expression of the chloroplast genes that led to the disappearance of the photosystem complexes, mimicking almost total photoinhibition. The treated plants, despite displaying only some early signs of senescence, sustained their metabolism and growth for several weeks. The only major remaining membrane component was LHCII antenna that formed superstructures - stacks of dozens of thylakoids or supergrana. Freeze-fracture electron microscopy revealed specific organization, directly displaying frequently bifurcated membranes with reduced or totally absent photosystem II (PSII) reaction centre complexes. Our findings show that it is possible to accumulate large amounts of light-harvesting membranes, organized into three-dimensional structures, in the absence of reaction centre complexes. This points to the reciprocal role of LHCII and PSII in self-assembly of the three-dimensional matrix of the photosynthetic membrane, dictating its size and flexible adaptation to the light environment.

Triplet Oxygen Evolution Catalyzed by a Biomimetic Oxomanganese Complex: Functional Role of the Carboxylate Buffer

Photosynthetic oxygen evolution involves water splitting into triplet oxygen, protons, and electrons, as follows: 2H2O → 3O2 + 4e–+ 4H+. The reaction is catalyzed by the oxomanganese complex of photosystem II, embedded in the thylakoid membrane of green plant chloroplasts and internal membranes of cyanobacteria. Biomimetic synthetic complexes have been developed over the years, although the reactivity of most of these complexes remains to be established. Here, we report a computational study of water splitting catalyzed by the mixed-valent oxomanganese dimer [H2O(terpy)MnIII(μ-oxo)2MnIV(terpy)OH2]3+ (terpy = 2,2′:6′,2″-terpyridine) in acetate buffer, with emphasis on the origin of triplet oxygen and the noninnocent role of carboxylate ligands in the underlying reaction mechanism. Our calculations suggest triplet oxygen evolution from an end-on (η1) Mn(III)-superoxo species, which forms from a hydroperoxo intermediate generated by nucleophilic attack of substrate water onto an oxyl radical Mn(IV)–O•. Carboxylate groups of acetate facilitate formation of the oxyl radical by shifting the redox potential of the Mn complex upon exchange with water ligands, and catalyze the O–O bond formation by deprotonating the nucleophilic water molecule. These findings provide valuable insights on the origin of triplet oxygen and on the regulatory role of the environment surrounding the inorganic core of oxomanganese complexes during catalytic oxygen evolution.

Artificial chloroplast: Au/chloroplast-morph-TiO2 with fast electron transfer and enhanced photocatalytic activity

The unique structures and functional features of chloroplasts in green plants provide a promising blueprint for greatly improving solar energy utilization efficiency. In this paper, a prototype of artificial chloroplast, Au/chloroplast-morph-TiO2 with natural chloroplasts' nanostructure and analogous functional features, is provided. The nano-layered structures of chloroplast template inherited in the chloroplast-morph-TiO2 lead to a large reaction area and fast photo-induced electron transfer; cocatalyst Au nanoparticles which work as reaction centers promote photo-induced charge separation and improve the overall photocatalytic activity in hydrogen production; Nitrogen and phosphorus self-doped from the bio-template increase visible light absorption, similar to the antenna pigment. With this new inorganic artificial photosynthetic system, we achieve effective light utilization, fast photo-induced charge separation, high electron transfer, enhanced photocatalytic activity for dye degradation rate and improved H2 evolution efficiency. This concept provides the inspiration for constructing efficient photocatalysts by imitating the photosynthesis process from both structures and functions.

Structural diversity of signal recognition particle RNAs in plastids

One of the pathways for protein targeting to the plasma membrane in bacteria utilizes the co-translationally acting signal recognition particle (SRP), a universally conserved ribonucleoprotein complex consisting of a 54 kDa protein and a functional RNA. An interesting exception is the higher plant chloroplast SRP, which lacks the otherwise essential RNA component. Furthermore, green plant chloroplasts have an additional post-translational SRP-dependent transport system in which the chloroplast-specific cpSRP43 protein binds to imported substrate proteins and to the conserved 54 kDa SRP subunit (cpSRP54). While homologs to the bacterial SRP protein and RNA component previously have been identified in genome sequences of red algae and diatoms, a recent study investigated the evolution of the green plant SRP system.1 Analysis of hundreds of plastid and nuclear genomes showed a surprising pattern of multiple losses of the plastid SRP RNA during evolution and a widespread presence in all non-spermatophyte plants and green algae. Contrary to expectations, all green organisms that have an identified cpSRP RNA also contain a cpSRP43. Notably, the structure of the plastid SRP RNAs is much more diverse than that of bacterial SRP RNAs. The apical GNRA tetraloop is only conserved in organisms of the red lineage and basal organisms of the green lineage, whereas further chloroplast SRP RNAs are characterized by atypical, mostly enlarged apical loops.

A 20-state empirical amino-acid substitution model for green plant chloroplasts

We introduce a new empirical amino-acid substitution model intended for use with for green plant (Viridiplantae) chloroplast protein data, which we call gcpREV (green chloroplast reversible model). Relative exchange rates and base composition frequencies among amino-acids were calculated using a Markov-chain Monte Carlo analysis on a combined data set of 34 proteins sampled among 27 taxa of green plants. The gcpREV model is a better fit than the commonly-used cpREV model to five previously published chloroplast protein test data sets of green plants, but is not a better fit to test data consisting mostly of non-green plant chloroplasts. Consequently, our analyses suggest a degree of specificity of the new model to green plant chloroplast data. The new model is implemented in the software P4, and model description files are available for other popular phylogenetic analysis software.

Oxidative Folding in Chloroplasts

Disulfide-bonded proteins in chloroplasts from green plants exist in the envelope and the thylakoid membrane, and in the stroma and the lumen. The formation of disulfide bonds in proteins is referred to as oxidative folding and is linked to the import and folding of chloroplast proteins as well as the assembly and repair of thylakoid complexes. It is also important in the redox regulation of enzymes and signal transfer. Green-plant chloroplasts contain enzymes that can form and isomerize disulfide bonds in proteins. In Arabidopsis thaliana, four proteins are identified that are relevant for the catalysis of disulfide bond formation in chloroplast proteins. The proteins' low quantum yield of Photosystem II 1 (LQY1, At1g75690) and snowy cotyledon 2 (SCO2, At3g19220) exhibits protein disulfide isomerase activity and is suggested to function in the assembly and repair of Photosystem II (PSII), and the biogenesis of thylakoids in cotyledons, respectively. The thylakoid-located Lumen thiol oxidoreductase 1 (LTO1, At4g35760) can catalyze the formation of the disulfide bond of the extrinsic PsbO protein of PSII. In addition, the stroma-located protein disulfide isomerase PDIL1-3 (At3g54960) may have a role in oxidative folding. Research on oxidative folding in chloroplasts plants is in an early stage and little is known about the mechanisms of disulfide bond formation in chloroplast proteins. The close link between the import and folding of chloroplast proteins suggests that Hsp93, a component of the inner envelope's import apparatus, may have co-chaperones that can catalyze disulfide bond formation in newly imported proteins.

Large Photovoltages Generated by Plant Photosystem I Crystals

Light energy conversion to electrical energy in photosynthesis and in photovoltaic devices is initiated by separation of charges that generates a photovoltage across an energy barrier in the system. [ 1–3 ] In conventional devices, mostly based on semiconductors, [ 4 ] the photovoltage is limited by the bandgap energy. Anomalous photovoltaic effects, in which the generated photovoltage is larger than the bandgap, have been reported in polar materials such as ferroelectric crystals, [ 5 , 6 ] but also in various amorphous and polycrystalline semiconductors. [ 7–10 ] Can a photoactive biological protein also generate an anomalous photovoltage? Natural candidates are photosynthetic reaction centers such as plant photosystem I (PSI), which are highly effi cient light energy converters found in thylakoid membranes inside the chloroplasts of green plants. [ 11 , 12 ] Here we show that these dried micrometer-thick crystalline biological protein complexes made of plant PSI [ 13 ] can generate unprecedented high photovoltages (well above 10 V) when placed on a conducting solid surface and measured by Kelvin probe force microscopy (KPFM). The measured photovoltages give rise to internal electric fi elds as large as 100 kV cm − 1 , which are among the highest values ever reported in an inorganic material system. We suggest that these high values result from depletion of the P700 electron donor centers and trapping centers in the PSI complex. We also present an equation depicting the dependence of the large photovoltage phenomenon in plant PSI crystals on light intensity. The unprecedented photovoltage, the microscale dimensions, and the high quantum yield make the PSI crystals intriguing units for potential use in optoelectronic devices. Photosynthetic reaction centers such as PSI evolved over a billion years to become highly effi cient light energy converters.

Plants

Plants

Analysis of high temperature stress on the dynamics of antenna size and reducing side heterogeneity of Photosystem II in wheat leaves ( Triticum aestivum)

This study demonstrates the effect of high temperature stress on the heterogeneous behavior of PSII in Wheat ( Triticum aestivum) leaves. Photosystem II in green plant chloroplasts displays heterogeneity both in the composition of its light harvesting antenna i.e. on the basis of antenna size (α, β and γ centers) and in the ability to reduce the plastoquinone pool i.e. the reducing side of the reaction centers (Q B-reducing centers and Q B-non-reducing centers). Detached wheat leaves were subjected to high temperature stress of 35 °C, 40 °C and 45 °C. The chlorophyll a (Chl a) fluorescence transient were recorded in vivo with high time resolution and analyzed according to JIP test which can quantify PS II behavior using Plant efficiency analyzer (PEA). Other than PEA, Biolyzer HP-3 software was used to evaluate different types of heterogeneity in wheat leaves. The results revealed that at high temperature, there was a change in the relative amounts of PSII α, β and γ centers. As judged from the complementary area growth curve, it seemed that with increasing temperature the PSII β and PSII γ centers increased at the expense of PSII α centers. The reducing side heterogeneity was also affected as shown by an increase in the number of Q B-non-reducing centers at high temperatures. The reversibility of high temperature induced damage on PSII heterogeneity was also studied. Antenna size heterogeneity was recovered fully up to 40 °C while reducing side heterogeneity showed partial recovery at 40 °C. An irreversible damage to both the types of heterogeneity was observed at 45 °C. The work is a significant contribution to understand the basic mechanism involved in the adaptation of crop plants to stress conditions.

Cytokinin-binding protein (70 kDa) from etioplasts and amyloplasts of etiolated maize seedlings and chloroplasts of green plants and its putative function

Cytokinins regulate chloroplast differentiation and functioning, but their targets in plastids are not known. In this connection, the plastid localization of the 70 kDa cytokinin-binding protein (CBP70) was studied immunocytochemically in 4-d-old etiolated maize seedlings (Zea mays L., cv. Elbrus) using monoclonal antibodies (mAbs) against CBP70 recognizing this protein not only in nuclei and cytoplasm, but also in plastids. CBP70 was detected in the amyloplasts of the root cap and etioplasts of the mesocotyl, stem apex, and leaves encircling the stem axis in the node. Immunogold electron microscopy demonstrated CBP70 localization in amyloplasts outside starch grains and revealed a dependence of CBP70 content in etioplasts on the degree of their inner membrane differentiation: the low CBP70 amount in etioplasts at the early stages of membrane development, the high content in etioplasts with actively developing membranes, and a considerable decrease in plastids with the formed prolamellar body. This suggests that CBP70 is involved in etioplast structure development. CBP70 was also observed in chloroplasts of the bundle sheath of green maize leaves. CBP70 purified from etioplasts mediated trans-zeatin-dependent activation of transcription elongation in vitro in the transcription systems of maize etioplasts and barley chloroplasts, suggesting that CBP70 is a plastid transcription elongation factor or a modulator of plastid elongation factor activity. CBP70 involvement in the cytokinin-dependent regulation of plastid transcription elongation could be essential for the cytokinin control of the biogenesis of this organelle.