33 results on '"Bock Ralph"'
Search Results
2. Characterization of mutants deficient in N-terminal phosphorylation of the chloroplast ATP synthase subunit β.
- Author
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Strand DD, Karcher D, Ruf S, Schadach A, Schöttler MA, Sandoval-Ibañez O, Hall D, Kramer DM, and Bock R
- Subjects
- Phosphorylation, Electron Transport, Adenosine Triphosphate metabolism, Chloroplast Proton-Translocating ATPases genetics, Chloroplast Proton-Translocating ATPases metabolism, Photosynthesis genetics
- Abstract
Understanding the regulation of photosynthetic light harvesting and electron transfer is of great importance to efforts to improve the ability of the electron transport chain to supply downstream metabolism. A central regulator of the electron transport chain is ATP synthase, the molecular motor that harnesses the chemiosmotic potential generated from proton-coupled electron transport to synthesize ATP. ATP synthase is regulated both thermodynamically and post-translationally, with proposed phosphorylation sites on multiple subunits. In this study we focused on two N-terminal serines on the catalytic subunit β in tobacco (Nicotiana tabacum), previously proposed to be important for dark inactivation of the complex to avoid ATP hydrolysis at night. Here we show that there is no clear role for phosphorylation in the dark inactivation of ATP synthase. Instead, mutation of one of the two phosphorylated serine residues to aspartate to mimic constitutive phosphorylation strongly decreased ATP synthase abundance. We propose that the loss of N-terminal phosphorylation of ATPβ may be involved in proper ATP synthase accumulation during complex assembly., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2023. Published by Oxford University Press on behalf of American Society of Plant Biologists.)
- Published
- 2023
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3. A transcriptional regulator that boosts grain yields and shortens the growth duration of rice.
- Author
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Wei S, Li X, Lu Z, Zhang H, Ye X, Zhou Y, Li J, Yan Y, Pei H, Duan F, Wang D, Chen S, Wang P, Zhang C, Shang L, Zhou Y, Yan P, Zhao M, Huang J, Bock R, Qian Q, and Zhou W
- Subjects
- Nitrogen metabolism, Transcription, Genetic, Crop Production, Edible Grain genetics, Edible Grain growth & development, Oryza genetics, Oryza growth & development, Photosynthesis genetics, Plant Proteins genetics, Plant Proteins metabolism, Transcription Factors genetics, Transcription Factors metabolism
- Abstract
Complex biological processes such as plant growth and development are often under the control of transcription factors that regulate the expression of large sets of genes and activate subordinate transcription factors in a cascade-like fashion. Here, by screening candidate photosynthesis-related transcription factors in rice, we identified a DREB (Dehydration Responsive Element Binding) family member, OsDREB1C, in which expression is induced by both light and low nitrogen status. We show that OsDREB1C drives functionally diverse transcriptional programs determining photosynthetic capacity, nitrogen utilization, and flowering time. Field trials with OsDREB1C -overexpressing rice revealed yield increases of 41.3 to 68.3% and, in addition, shortened growth duration, improved nitrogen use efficiency, and promoted efficient resource allocation, thus providing a strategy toward achieving much-needed increases in agricultural productivity.
- Published
- 2022
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4. A photosynthesis operon in the chloroplast genome drives speciation in evening primroses.
- Author
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Zupok A, Kozul D, Schöttler MA, Niehörster J, Garbsch F, Liere K, Fischer A, Zoschke R, Malinova I, Bock R, and Greiner S
- Subjects
- Acclimatization genetics, Cytochrome b6f Complex genetics, Light, Oenothera biennis physiology, Photosystem II Protein Complex genetics, Plant Proteins genetics, Plastids genetics, Promoter Regions, Genetic, RNA Editing, Genetic Speciation, Genome, Chloroplast, Oenothera biennis genetics, Operon, Photosynthesis genetics
- Abstract
Genetic incompatibility between the cytoplasm and the nucleus is thought to be a major factor in species formation, but mechanistic understanding of this process is poor. In evening primroses (Oenothera spp.), a model plant for organelle genetics and population biology, hybrid offspring regularly display chloroplast-nuclear incompatibility. This usually manifests in bleached plants, more rarely in hybrid sterility or embryonic lethality. Hence, most of these incompatibilities affect photosynthetic capability, a trait that is under selection in changing environments. Here we show that light-dependent misregulation of the plastid psbB operon, which encodes core subunits of photosystem II and the cytochrome b6f complex, can lead to hybrid incompatibility, and this ultimately drives speciation. This misregulation causes an impaired light acclimation response in incompatible plants. Moreover, as a result of their different chloroplast genotypes, the parental lines differ in photosynthesis performance upon exposure to different light conditions. Significantly, the incompatible chloroplast genome is naturally found in xeric habitats with high light intensities, whereas the compatible one is limited to mesic habitats. Consequently, our data raise the possibility that the hybridization barrier evolved as a result of adaptation to specific climatic conditions., (© The Author(s) 2021. Published by Oxford University Press on behalf of American Society of Plant Biologists.)
- Published
- 2021
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5. Photosynthesis without β-carotene.
- Author
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Xu P, Chukhutsina VU, Nawrocki WJ, Schansker G, Bielczynski LW, Lu Y, Karcher D, Bock R, and Croce R
- Subjects
- Plants, Genetically Modified metabolism, Plants, Genetically Modified physiology, Nicotiana metabolism, Nicotiana physiology, Xanthophylls metabolism, beta Carotene metabolism, Photosynthesis, beta Carotene physiology
- Abstract
Carotenoids are essential in oxygenic photosynthesis: they stabilize the pigment-protein complexes, are active in harvesting sunlight and in photoprotection. In plants, they are present as carotenes and their oxygenated derivatives, xanthophylls. While mutant plants lacking xanthophylls are capable of photoautotrophic growth, no plants without carotenes in their photosystems have been reported so far, which has led to the common opinion that carotenes are essential for photosynthesis. Here, we report the first plant that grows photoautotrophically in the absence of carotenes: a tobacco plant containing only the xanthophyll astaxanthin. Surprisingly, both photosystems are fully functional despite their carotenoid-binding sites being occupied by astaxanthin instead of β-carotene or remaining empty (i.e. are not occupied by carotenoids). These plants display non-photochemical quenching, despite the absence of both zeaxanthin and lutein and show that tobacco can regulate the ratio between the two photosystems in a very large dynamic range to optimize electron transport., Competing Interests: PX, VC, WN, GS, LB, YL, DK, RB, RC No competing interests declared, (© 2020, Xu et al.)
- Published
- 2020
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6. Expression of a carotenogenic gene allows faster biomass production by redesigning plant architecture and improving photosynthetic efficiency in tobacco.
- Author
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Moreno JC, Mi J, Agrawal S, Kössler S, Turečková V, Tarkowská D, Thiele W, Al-Babili S, Bock R, and Schöttler MA
- Subjects
- Abscisic Acid metabolism, Daucus carota genetics, Gene Expression Regulation, Plant genetics, Genes, Plant genetics, Gibberellins metabolism, Metabolic Networks and Pathways genetics, Photosynthesis physiology, Plant Growth Regulators metabolism, Plant Proteins genetics, Plant Proteins physiology, Plants, Genetically Modified, Nicotiana anatomy & histology, Nicotiana metabolism, Nicotiana physiology, Up-Regulation, Carotenoids metabolism, Genes, Plant physiology, Photosynthesis genetics, Nicotiana growth & development
- Abstract
Because carotenoids act as accessory pigments in photosynthesis, play a key photoprotective role and are of major nutritional importance, carotenogenesis has been a target for crop improvement. Although carotenoids are important precursors of phytohormones, previous genetic manipulations reported little if any effects on biomass production and plant development, but resulted in specific modifications in carotenoid content. Unexpectedly, the expression of the carrot lycopene β-cyclase (DcLCYB1) in Nicotiana tabacum cv. Xanthi not only resulted in increased carotenoid accumulation, but also in altered plant architecture characterized by longer internodes, faster plant growth, early flowering and increased biomass. Here, we have challenged these transformants with a range of growth conditions to determine the robustness of their phenotype and analyze the underlying mechanisms. Transgenic DcLCYB1 lines showed increased transcript levels of key genes involved in carotenoid, chlorophyll, gibberellin (GA) and abscisic acid (ABA) biosynthesis, but also in photosynthesis-related genes. Accordingly, their carotenoid, chlorophyll, ABA and GA contents were increased. Hormone application and inhibitor experiments confirmed the key role of altered GA/ABA contents in the growth phenotype. Because the longer internodes reduce shading of mature leaves, induction of leaf senescence was delayed, and mature leaves maintained a high photosynthetic capacity. This increased total plant assimilation, as reflected in higher plant yields under both fully controlled constant and fluctuating light, and in non-controlled conditions. Furthermore, our data are a warning that engineering of isoprenoid metabolism can cause complex changes in phytohormone homeostasis and therefore plant development, which have not been sufficiently considered in previous studies., (© 2020 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)
- Published
- 2020
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7. Redesigning photosynthesis to sustainably meet global food and bioenergy demand.
- Author
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Ort DR, Merchant SS, Alric J, Barkan A, Blankenship RE, Bock R, Croce R, Hanson MR, Hibberd JM, Long SP, Moore TA, Moroney J, Niyogi KK, Parry MA, Peralta-Yahya PP, Prince RC, Redding KE, Spalding MH, van Wijk KJ, Vermaas WF, von Caemmerer S, Weber AP, Yeates TO, Yuan JS, and Zhu XG
- Subjects
- Biofuels, Crops, Agricultural physiology, Food Supply, Photosynthesis
- Abstract
The world's crop productivity is stagnating whereas population growth, rising affluence, and mandates for biofuels put increasing demands on agriculture. Meanwhile, demand for increasing cropland competes with equally crucial global sustainability and environmental protection needs. Addressing this looming agricultural crisis will be one of our greatest scientific challenges in the coming decades, and success will require substantial improvements at many levels. We assert that increasing the efficiency and productivity of photosynthesis in crop plants will be essential if this grand challenge is to be met. Here, we explore an array of prospective redesigns of plant systems at various scales, all aimed at increasing crop yields through improved photosynthetic efficiency and performance. Prospects range from straightforward alterations, already supported by preliminary evidence of feasibility, to substantial redesigns that are currently only conceptual, but that may be enabled by new developments in synthetic biology. Although some proposed redesigns are certain to face obstacles that will require alternate routes, the efforts should lead to new discoveries and technical advances with important impacts on the global problem of crop productivity and bioenergy production.
- Published
- 2015
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8. Photosynthetic Membranes of Synechocystis or Plants Convert Sunlight to Photocurrent through Different Pathways due to Different Architectures.
- Author
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Pinhassi RI, Kallmann D, Saper G, Larom S, Linkov A, Boulouis A, Schöttler MA, Bock R, Rothschild A, Adir N, and Schuster G
- Subjects
- Electrodes, Electron Transport physiology, Electrons, Light, Sunlight, Synechocystis metabolism, Thylakoids metabolism, Photosynthesis physiology, Photosystem II Protein Complex metabolism, Synechocystis physiology, Thylakoids physiology
- Abstract
Thylakoid membranes contain the redox active complexes catalyzing the light-dependent reactions of photosynthesis in cyanobacteria, algae and plants. Crude thylakoid membranes or purified photosystems from different organisms have previously been utilized for generation of electrical power and/or fuels. Here we investigate the electron transferability from thylakoid preparations from plants or the cyanobacterium Synechocystis. We show that upon illumination, crude Synechocystis thylakoids can reduce cytochrome c. In addition, this crude preparation can transfer electrons to a graphite electrode, producing an unmediated photocurrent of 15 μA/cm2. Photocurrent could be obtained in the presence of the PSII inhibitor DCMU, indicating that the source of electrons is QA, the primary Photosystem II acceptor. In contrast, thylakoids purified from plants could not reduce cyt c, nor produced a photocurrent in the photocell in the presence of DCMU. The production of significant photocurrent (100 μA/cm2) from plant thylakoids required the addition of the soluble electron mediator DCBQ. Furthermore, we demonstrate that use of crude thylakoids from the D1-K238E mutant in Synechocystis resulted in improved electron transferability, increasing the direct photocurrent to 35 μA/cm2. Applying the analogous mutation to tobacco plants did not achieve an equivalent effect. While electron abstraction from crude thylakoids of cyanobacteria or plants is feasible, we conclude that the site of the abstraction of the electrons from the thylakoids, the architecture of the thylakoid preparations influence the site of the electron abstraction, as well as the transfer pathway to the electrode. This dictates the use of different strategies for production of sustainable electrical current from photosynthetic thylakoid membranes of cyanobacteria or higher plants.
- Published
- 2015
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9. Tomato fruit photosynthesis is seemingly unimportant in primary metabolism and ripening but plays a considerable role in seed development.
- Author
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Lytovchenko A, Eickmeier I, Pons C, Osorio S, Szecowka M, Lehmberg K, Arrivault S, Tohge T, Pineda B, Anton MT, Hedtke B, Lu Y, Fisahn J, Bock R, Stitt M, Grimm B, Granell A, and Fernie AR
- Subjects
- Aminolevulinic Acid metabolism, Fruit genetics, Fruit metabolism, Fruit physiology, Gene Expression Profiling, Gene Expression Regulation, Plant physiology, Glucuronidase, Solanum lycopersicum genetics, Solanum lycopersicum metabolism, Oligonucleotide Array Sequence Analysis, Organ Specificity, Phenotype, Plant Proteins genetics, Plants, Genetically Modified, Promoter Regions, Genetic genetics, Reproduction, Seeds genetics, Seeds metabolism, Fruit growth & development, Solanum lycopersicum growth & development, Photosynthesis physiology, Plant Proteins metabolism, Seeds growth & development
- Abstract
Fruit of tomato (Solanum lycopersicum), like those from many species, have been characterized to undergo a shift from partially photosynthetic to truly heterotrophic metabolism. While there is plentiful evidence for functional photosynthesis in young tomato fruit, the rates of carbon assimilation rarely exceed those of carbon dioxide release, raising the question of its role in this tissue. Here, we describe the generation and characterization of lines exhibiting a fruit-specific reduction in the expression of glutamate 1-semialdehyde aminotransferase (GSA). Despite the fact that these plants contained less GSA protein and lowered chlorophyll levels and photosynthetic activity, they were characterized by few other differences. Indeed, they displayed almost no differences in fruit size, weight, or ripening capacity and furthermore displayed few alterations in other primary or intermediary metabolites. Although GSA antisense lines were characterized by significant alterations in the expression of genes associated with photosynthesis, as well as with cell wall and amino acid metabolism, these changes were not manifested at the phenotypic level. One striking feature of the antisense plants was their seed phenotype: the transformants displayed a reduced seed set and altered morphology and metabolism at early stages of fruit development, although these differences did not affect the final seed number or fecundity. Taken together, these results suggest that fruit photosynthesis is, at least under ambient conditions, not necessary for fruit energy metabolism or development but is essential for properly timed seed development and therefore may confer an advantage under conditions of stress.
- Published
- 2011
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10. ATP synthase repression in tobacco restricts photosynthetic electron transport, CO2 assimilation, and plant growth by overacidification of the thylakoid lumen.
- Author
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Rott M, Martins NF, Thiele W, Lein W, Bock R, Kramer DM, and Schöttler MA
- Subjects
- Antisense Elements (Genetics), Plant Proteins metabolism, Plants, Genetically Modified enzymology, Plants, Genetically Modified growth & development, Point Mutation, Nicotiana growth & development, Transformation, Genetic, ATP Synthetase Complexes metabolism, Carbon Dioxide metabolism, Electron Transport, Photosynthesis, Thylakoids enzymology, Nicotiana enzymology
- Abstract
Tobacco (Nicotiana tabacum) plants strictly adjust the contents of both ATP synthase and cytochrome b(6)f complex to the metabolic demand for ATP and NADPH. While the cytochrome b(6)f complex catalyzes the rate-limiting step of photosynthetic electron flux and thereby controls assimilation, the functional significance of the ATP synthase adjustment is unknown. Here, we reduced ATP synthase accumulation by an antisense approach directed against the essential nuclear-encoded γ-subunit (AtpC) and by the introduction of point mutations into the translation initiation codon of the plastid-encoded atpB gene (encoding the essential β-subunit) via chloroplast transformation. Both strategies yielded transformants with ATP synthase contents ranging from 100 to <10% of wild-type levels. While the accumulation of the components of the linear electron transport chain was largely unaltered, linear electron flux was strongly inhibited due to decreased rates of plastoquinol reoxidation at the cytochrome b(6)f complex (photosynthetic control). Also, nonphotochemical quenching was triggered at very low light intensities, strongly reducing the quantum efficiency of CO(2) fixation. We show evidence that this is due to an increased steady state proton motive force, resulting in strong lumen overacidification, which in turn represses photosynthesis due to photosynthetic control and dissipation of excitation energy in the antenna bed.
- Published
- 2011
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11. RBF1, a Plant Homolog of the Bacterial Ribosome-Binding Factor RbfA, Acts in Processing of the Chloroplast 16S Ribosomal RNA
- Author
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Fristedt, Rikard, Scharff, Lars B, Clarke, Cornelia A, Wang, Qin, Lin, Chentao, Merchant, Sabeeha S, and Bock, Ralph
- Subjects
Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Amino Acid Sequence ,Arabidopsis ,Arabidopsis Proteins ,Chlamydomonas reinhardtii ,Chloroplasts ,Escherichia coli Proteins ,Genetic Complementation Test ,Molecular Sequence Data ,Mutation ,Photosynthesis ,Plants ,Genetically Modified ,Plastids ,RNA ,Ribosomal ,16S ,Ribosomal Proteins ,Sequence Homology ,Amino Acid ,Thylakoids ,Biological Sciences ,Agricultural and Veterinary Sciences ,Plant Biology & Botany - Abstract
Plastids (chloroplasts) possess 70S ribosomes that are very similar in structure and function to the ribosomes of their bacterial ancestors. While most components of the bacterial ribosome (ribosomal RNAs [rRNAs] and ribosomal proteins) are well conserved in the plastid ribosome, little is known about the factors mediating the biogenesis of plastid ribosomes. Here, we have investigated a putative homolog of the bacterial RbfA (for ribosome-binding factor A) protein that was identified as a cold-shock protein and an auxiliary factor acting in the 5' maturation of the 16S rRNA. The unicellular green alga Chlamydomonas reinhardtii and the vascular plant Arabidopsis (Arabidopsis thaliana) both encode a single RbfA-like protein in their nuclear genomes. By generating specific antibodies against this protein, we show that the plant RbfA-like protein functions exclusively in the plastid, where it is associated with thylakoid membranes. Analysis of mutants for the corresponding gene (termed RBF1) reveals that the gene function is essential for photoautotrophic growth. Weak mutant alleles display reduced levels of plastid ribosomes, a specific depletion in 30S ribosomal subunits, and reduced activity of plastid protein biosynthesis. Our data suggest that, while the function in ribosome maturation and 16S rRNA 5' end processing is conserved, the RBF1 protein has assumed an additional role in 3' end processing. Together with the apparent absence of a homologous protein from plant mitochondria, our findings illustrate that the assembly process of the 70S ribosome is not strictly conserved and has undergone some modifications during organelle evolution.
- Published
- 2014
12. The plastid-encoded PsaI subunit stabilizes photosystem I during leaf senescence in tobacco
- Author
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Schöttler, Mark Aurel, Thiele, Wolfram, Belkius, Karolina, Bergner, Sonja Verena, Flügel, Claudia, Wittenberg, Gal, Agrawal, Shreya, Stegemann, Sandra, Ruf, Stephanie, and Bock, Ralph
- Published
- 2017
13. Generation and characterization of a collection of knock-down lines for the chloroplast Clp protease complex in tobacco
- Author
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Moreno, Juan C., Tiller, Nadine, Diez, Mercedes, Karcher, Daniel, Tillich, Michael, Schöttler, Mark A., and Bock, Ralph
- Published
- 2017
14. A Small Chloroplast-Encoded Protein as a Novel Architectural Component of the Light-Harvesting Antenna
- Author
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Ruf, Stephanie, Biehler, Klaus, and Bock, Ralph
- Published
- 2000
15. Transfer of the cytochrome P450-dependent dhurrin pathway from Sorghum bicolor into Nicotiana tabacum chloroplasts for light-driven synthesis
- Author
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Gnanasekaran, Thiyagarajan, Karcher, Daniel, Nielsen, Agnieszka Zygadlo, Martens, Helle Juel, Ruf, Stephanie, Kroop, Xenia, Olsen, Carl Erik, Motawie, Mohammed Saddik, Pribil, Mathias, Møller, Birger Lindberg, Bock, Ralph, and Jensen, Poul Erik
- Published
- 2016
16. Systems Analysis of the Response of Photosynthesis, Metabolism, and Growth to an Increase in Irradiance in the Photosynthetic Model Organism Chlamydomonas reinhardtii
- Author
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Mettler, Tabea, Mühlhaus, Timo, Hemme, Dorothea, Schöttler, Mark-Aurel, Rupprecht, Jens, Idoine, Adam, Veyel, Daniel, Pal, Sunil Kumar, Yaneva-Roder, Liliya, Winck, Flavia Vischi, Sommer, Frederik, Vosloh, Daniel, Seiwert, Bettina, Erban, Alexander, Burgos, Asdrubal, Arvidsson, Samuel, Schönfelder, Stephanie, Arnold, Anne, Günther, Manuela, Krause, Ursula, Lohse, Marc, Kopka, Joachim, Nikoloski, Zoran, Mueller-Roeber, Bernd, Willmitzer, Lothar, Bock, Ralph, Schroda, Michael, and Stitt, Mark
- Published
- 2014
17. Targeted inactivation of the smallest plastid genome‐encoded open reading frame reveals a novel and essential subunit of the cytochrome b6f complex
- Author
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Hager, Martin, Biehler, Klaus, Illerhaus, Jürgen, Ruf, Stephanie, and Bock, Ralph
- Published
- 1999
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18. Highly Resolved Systems Biology to Dissect the Etioplast-to-Chloroplast Transition in Tobacco Leaves1[OPEN]
- Author
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Armarego-Marriott, Tegan, Kowalewska, Łucja, Burgos, Asdrubal, Fischer, Axel, Thiele, Wolfram, Erban, Alexander, Strand, Deserah, Kahlau, Sabine, Hertle, Alexander, Kopka, Joachim, Walther, Dirk, Reich, Ziv, Schöttler, Mark Aurel, and Bock, Ralph
- Subjects
Cell Nucleus ,Chloroplasts ,Light ,Systems Biology ,fungi ,Genome, Plastid ,food and beverages ,Articles ,Lipid Metabolism ,Plant Leaves ,Microscopy, Electron, Transmission ,Tobacco ,Carbohydrate Metabolism ,sense organs ,Plastids ,Amino Acids ,Photosynthesis ,Transcriptome ,Triglycerides - Abstract
Upon exposure to light, plant cells quickly acquire photosynthetic competence by converting pale etioplasts into green chloroplasts. This developmental transition involves the de novo biogenesis of the thylakoid system and requires reprogramming of metabolism and gene expression. Etioplast-to-chloroplast differentiation involves massive changes in plastid ultrastructure, but how these changes are connected to specific changes in physiology, metabolism, and expression of the plastid and nuclear genomes is poorly understood. Here, we describe a new experimental system in the dicotyledonous model plant tobacco (Nicotiana tabacum) that allows us to study the leaf deetiolation process at the systems level. We have determined the accumulation kinetics of photosynthetic complexes, pigments, lipids, and soluble metabolites and recorded the dynamic changes in plastid ultrastructure and in the nuclear and plastid transcriptomes. Our data describe the greening process at high temporal resolution, resolve distinct genetic and metabolic phases during deetiolation, and reveal numerous candidate genes that may be involved in light-induced chloroplast development and thylakoid biogenesis.
- Published
- 2019
19. Curvature thylakoid 1 proteins modulate prolamellar body morphology and promote organized thylakoid biogenesis in Arabidopsis thaliana.
- Author
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Sandoval-Ibáñez, Omar, Sharma, Anurag, Bykowski, Michał, Borràs-Gas, Guillem, Behrendorff, James B. Y. H., Mellor, Silas, Qvortrup, Klaus, Verdonk, Julian C., Bock, Ralph, Kowalewska, Łucja, and Pribil, Mathias
- Subjects
ARABIDOPSIS thaliana ,IMMOBILIZED proteins ,CURVATURE ,CHLOROPLAST membranes ,COMMERCIAL products ,PROTEINS - Abstract
The term "de-etiolation" refers to the light-dependent differentiation of etioplasts to chloroplasts in angiosperms. The underlying process involves reorganization of prolamellar bodies (PLBs) and prothylakoids into thylakoids, with concurrent changes in protein, lipid, and pigment composition, which together lead to the assembly of active photosynthetic complexes. Despite the highly conserved structure of PLBs among land plants, the processes that mediate PLB maintenance and their disassembly during de-etiolation are poorly understood. Among chloroplast thylakoid membrane--localized proteins, to date, only Curvature thylakoid 1 (CURT1) proteins were shown to exhibit intrinsic membrane-bending capacity. Here, we show that CURT1 proteins, which play a critical role in grana margin architecture and thylakoid plasticity, also participate in de-etiolation and modulate PLB geometry and density. Lack of CURT1 proteins severely perturbs PLB organization and vesicle fusion, leading to reduced accumulation of the light-dependent enzyme protochlorophyllide oxidoreductase (LPOR) and a delay in the onset of photosynthesis. In contrast, overexpression of CURT1A induces excessive bending of PLB membranes, which upon illumination show retarded disassembly and concomitant overaccumulation of LPOR, though without affecting greening or the establishment of photosynthesis. We conclude that CURT1 proteins contribute to the maintenance of the paracrystalline PLB morphology and are necessary for efficient and organized thylakoid membrane maturation during de-etiolation. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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20. Lycopene β-cyclase expression influences plant physiology, development, and metabolism in tobacco plants.
- Author
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Kössler, Stella, Armarego-Marriott, Tegan, Tarkowská, Danuše, Turečková, Veronika, Agrawal, Shreya, Mi, Jianing, Souza, Leonardo Perez de, Schöttler, Mark Aurel, Schadach, Anne, Fröhlich, Anja, Bock, Ralph, Al-Babili, Salim, Ruf, Stephanie, Sampathkumar, Arun, and Moreno, Juan C
- Subjects
PLANT physiology ,LYCOPENE ,PLANT metabolism ,PLANT biomass ,PLANT pigments ,CARROTS ,ABSCISIC acid ,TOBACCO - Abstract
Carotenoids are important isoprenoids produced in the plastids of photosynthetic organisms that play key roles in photoprotection and antioxidative processes. β-Carotene is generated from lycopene by lycopene β-cyclase (LCYB). Previously, we demonstrated that the introduction of the Daucus carota (carrot) DcLCYB1 gene into tobacco (cv. Xanthi) resulted in increased levels of abscisic acid (ABA) and especially gibberellins (GAs), resulting in increased plant yield. In order to understand this phenomenon prior to exporting this genetic strategy to crops, we generated tobacco (Nicotiana tabacum cv. Petit Havana) mutants that exhibited a wide range of LCYB expression. Transplastomic plants expressing DcLCYB1 at high levels showed a wild-type-like growth, even though their pigment content was increased and their leaf GA
1 content was reduced. RNA interference (RNAi) NtLCYB lines showed different reductions in NtLCYB transcript abundance, correlating with reduced pigment content and plant variegation. Photosynthesis (leaf absorptance, Fv / Fm , and light-saturated capacity of linear electron transport) and plant growth were impaired. Remarkably, drastic changes in phytohormone content also occurred in the RNAi lines. However, external application of phytohormones was not sufficient to rescue these phenotypes, suggesting that altered photosynthetic efficiency might be another important factor explaining their reduced biomass. These results show that LCYB expression influences plant biomass by different mechanisms and suggests thresholds for LCYB expression levels that might be beneficial or detrimental for plant growth. [ABSTRACT FROM AUTHOR]- Published
- 2021
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21. LCAA, a Novel Factor Required for Magnesium Protoporphyrin Monomethylester Cyclase Accumulation and Feedback Control of Aminolevulinic Acid Biosynthesis in Tobacco1[W][OA]
- Author
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Albus, Christin Anne, Salinas, Annabel, Czarnecki, Olaf, Kahlau, Sabine, Rothbart, Maxi, Thiele, Wolfram, Lein, Wolfgang, Bock, Ralph, Grimm, Bernhard, and Schöttler, Mark Aurel
- Subjects
Chlorophyll ,Feedback, Physiological ,Chlorophyll A ,Molecular Sequence Data ,Light-Harvesting Protein Complexes ,food and beverages ,Protoporphyrins ,Aminolevulinic Acid ,Fluorescence ,Evolution, Molecular ,Intramolecular Oxidoreductases ,Protein Transport ,Phenotype ,Tetrapyrroles ,Gene Expression Regulation, Plant ,Tobacco ,RNA, Antisense ,Amino Acid Sequence ,Plastids ,Photosynthesis ,Oxidation-Reduction ,Sequence Alignment ,Bioenergetics and Photosynthesis ,Conserved Sequence ,Plant Proteins - Abstract
Low Chlorophyll Accumulation A (LCAA) antisense plants were obtained from a screen for genes whose partial down-regulation results in a strong chlorophyll deficiency in tobacco (Nicotiana tabacum). The LCAA mutants are affected in a plastid-localized protein of unknown function, which is conserved in cyanobacteria and all photosynthetic eukaryotes. They suffer from drastically reduced light-harvesting complex (LHC) contents, while the accumulation of all other photosynthetic complexes per leaf area is less affected. As the disturbed accumulation of LHC proteins could be either attributable to a defect in LHC biogenesis itself or to a bottleneck in chlorophyll biosynthesis, chlorophyll synthesis rates and chlorophyll synthesis intermediates were measured. LCAA antisense plants accumulate magnesium (Mg) protoporphyrin monomethylester and contain reduced protochlorophyllide levels and a reduced content of CHL27, a subunit of the Mg protoporphyrin monomethylester cyclase. Bimolecular fluorescence complementation assays confirm a direct interaction between LCAA and CHL27. 5-Aminolevulinic acid synthesis rates are increased and correlate with an increased content of glutamyl-transfer RNA reductase. We suggest that LCAA encodes an additional subunit of the Mg protoporphyrin monomethylester cyclase, is required for the stability of CHL27, and contributes to feedback-control of 5-aminolevulinic acid biosynthesis, the rate-limiting step of chlorophyll biosynthesis.
- Published
- 2012
22. Identification and characterization of a stable intermediate in photosystem I assembly in tobacco.
- Author
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Wittenberg, Gal, Järvi, Sari, Hojka, Marta, Tóth, Szilvia Z., Meyer, Etienne H., Aro, Eva‐Mari, Schöttler, Mark A., and Bock, Ralph
- Subjects
PHOTOSYSTEMS ,TOBACCO ,MEMBRANE proteins ,PROSTHETIC groups (Enzymes) ,THYLAKOIDS - Abstract
Photosystem I ( PSI) is the most efficient bioenergetic nanomachine in nature and one of the largest membrane protein complexes known. It is composed of 18 protein subunits that bind more than 200 co-factors and prosthetic groups. While the structure and function of PSI have been studied in great detail, very little is known about the PSI assembly process. In this work, we have characterized a PSI assembly intermediate in tobacco plants, which we named PSI*. We found PSI* to contain only a specific subset of the core subunits of PSI. PSI* is particularly abundant in young leaves where active thylakoid biogenesis takes place. Moreover, PSI* was found to overaccumulate in PsaF-deficient mutant plants, and we show that re-initiation of PsaF synthesis promotes the maturation of PSI* into PSI. The attachment of antenna proteins to PSI also requires the transition from PSI* to mature PSI. Our data could provide a biochemical entry point into the challenging investigation of PSI biogenesis and allow us to improve the model for the assembly pathway of PSI in thylakoid membranes of vascular plants. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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23. Regulation of ascorbate biosynthesis in green algae has evolved to enable rapid stress-induced response via the VTC2 gene encoding GDP- l-galactose phosphorylase.
- Author
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Vidal‐Meireles, André, Neupert, Juliane, Zsigmond, Laura, Rosado‐Souza, Laise, Kovács, László, Nagy, Valéria, Galambos, Anikó, Fernie, Alisdair R., Bock, Ralph, and Tóth, Szilvia Z.
- Subjects
GREEN algae ,BIOSYNTHESIS ,GALACTOSE ,PHOSPHORYLASES ,VITAMIN C - Abstract
Ascorbate (vitamin C) plays essential roles in stress resistance, development, signaling, hormone biosynthesis and regulation of gene expression; however, little is known about its biosynthesis in algae., In order to provide experimental proof for the operation of the Smirnoff-Wheeler pathway described for higher plants and to gain more information on the regulation of ascorbate biosynthesis in Chlamydomonas reinhardtii, we targeted the VTC2 gene encoding GDP- l-galactose phosphorylase using artificial micro RNAs., Ascorbate concentrations in VTC2 ami RNA lines were reduced to 10% showing that GDP- l-galactose phosphorylase plays a pivotal role in ascorbate biosynthesis. The VTC2 ami RNA lines also grow more slowly, have lower chlorophyll content, and are more susceptible to stress than the control strains. We also demonstrate that: expression of the VTC2 gene is rapidly induced by H
2 O2 and1 O2 resulting in a manifold increase in ascorbate content; in contrast to plants, there is no circadian regulation of ascorbate biosynthesis; photosynthesis is not required per se for ascorbate biosynthesis; and Chlamydomonas VTC2 lacks negative feedback regulation by ascorbate in the physiological concentration range., Our work demonstrates that ascorbate biosynthesis is also highly regulated in Chlamydomonas albeit via mechanisms distinct from those previously described in land plants. [ABSTRACT FROM AUTHOR]- Published
- 2017
- Full Text
- View/download PDF
24. The Diurnal Logic of the Expression of the Chloroplast Genome in Chlamydomonas reinhardtii.
- Author
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Idoine, Adam D., Boulouis, Alix, Rupprecht, Jens, and Bock, Ralph
- Subjects
CHLOROPLASTS ,GENOMES ,CHLAMYDOMONAS reinhardtii ,GENE expression ,REACTIVE oxygen species - Abstract
Chloroplasts are derived from cyanobacteria and have retained a bacterial-type genome and gene expression machinery. The chloroplast genome encodes many of the core components of the photosynthetic apparatus in the thylakoid membranes. To avoid photooxidative damage and production of harmful reactive oxygen species (ROS) by incompletely assembled thylakoid protein complexes, chloroplast gene expression must be tightly regulated and co-ordinated with gene expression in the nucleus. Little is known about the control of chloroplast gene expression at the genome-wide level in response to internal rhythms and external cues. To obtain a comprehensive picture of organelle transcript levels in the unicellular model alga Chlamydomonas reinhardtii in diurnal conditions, a qRT-PCR platform was developed and used to quantify 68 chloroplast, 21 mitochondrial as well as 71 nuclear transcripts in cells grown in highly controlled 12 h light/12 h dark cycles. Interestingly, in anticipation of dusk, chloroplast transcripts from genes involved in transcription reached peak levels first, followed by transcripts from genes involved in translation, and finally photosynthesis gene transcripts. This pattern matches perfectly the theoretical demands of a cell “waking up” from the night. A similar trend was observed in the nuclear transcripts. These results suggest a striking internal logic in the expression of the chloroplast genome and a previously unappreciated complexity in the regulation of chloroplast genes. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
25. Reverse genetics in complex multigene operons by co-transformation of the plastid genome and its application to the open reading frame previously designated psbN.
- Author
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Krech, Katharina, Fu, Han‐Yi, Thiele, Wolfram, Ruf, Stephanie, Schöttler, Mark A., and Bock, Ralph
- Subjects
PLANT genetics ,PLASTIDS ,PLANT proteins ,PLANT mutation ,GENE expression in plants ,PHOTOSYNTHESIS ,GENETIC recombination ,PLANTS - Abstract
Reverse genetics approaches have contributed enormously to the elucidation of gene functions in plastid genomes and the determination of structure-function relationships in chloroplast multiprotein complexes. Gene knock-outs are usually performed by disrupting the reading frame of interest with a selectable marker cassette. Site-directed mutagenesis is done by placing the marker into the adjacent intergenic spacer and relying on co-integration of the desired mutation by homologous recombination. These strategies are not applicable to genes residing in large multigene operons or other gene-dense genomic regions, because insertion of the marker cassette into an operon-internal gene or into the nearest intergenic spacer is likely to interfere with expression of adjacent genes in the operon or disrupt cis-elements for the expression of neighboring genes and operons. Here we have explored the possibility of using a co-transformation strategy to mutate a small gene of unknown function ( psbN) that is embedded in a complex multigene operon. Although inactivation of psbN resulted in strong impairment of photosynthesis, homoplasmic knock-out lines were readily recovered by co-transformation with a selectable marker integrating >38 kb away from the targeted psbN. Our results suggest co-transformation as a suitable strategy for the functional analysis of plastid genes and operons, which allows the recovery of unselected homoplasmic mutants even if the introduced mutations entail a significant selective disadvantage. Moreover, our data provide evidence for involvement of the psbN gene product in the biogenesis of both photosystem I and photosystem II. We therefore propose to rename the gene product 'photosystem biogenesis factor 1′ and the gene pbf1. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
26. Design of chimeric expression elements that confer high-level gene activity in chromoplasts.
- Author
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Caroca, Rodrigo, Howell, Katharine A., Hasse, Claudia, Ruf, Stephanie, and Bock, Ralph
- Subjects
CHIMERISM ,GENE expression in plants ,CHROMOPLASTS ,PHOTOSYNTHESIS ,PLASTIDS ,PLANT genomes ,PLANT proteins - Abstract
Non-green plastids, such as chromoplasts, generally have much lower activity of gene expression than chloroplasts in photosynthetically active tissues. Suppression of plastid genes in non-green tissues occurs through a complex interplay of transcriptional and translational control, with the contribution of regulation of transcript abundance versus translational activity being highly variable between genes. Here, we have investigated whether the low expression of the plastid genome in chromoplasts results from inherent limitations in gene expression capacity, or can be overcome by designing appropriate combinations of promoters and translation initiation signals in the 5′ untranslated region (5′- UTR). We constructed chimeric expression elements that combine promoters and 5′- UTRs from plastid genes, which are suppressed during chloroplast-to-chromoplast conversion in Solanum lycopersicum (tomato) fruit ripening, either just at the translational level or just at the level of m RNA accumulation. These chimeric expression elements were introduced into the tomato plastid genome by stable chloroplast transformation. We report the identification of promoter- UTR combinations that confer high-level gene expression in chromoplasts of ripe tomato fruits, resulting in the accumulation of reporter protein GFP to up to 1% of total cellular protein. Our work demonstrates that non-green plastids are capable of expressing genes to high levels. Moreover, the chimeric cis-elements for chromoplasts developed here are widely applicable in basic and applied research using transplastomic methods. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
27. Identification of protein stability determinants in chloroplasts.
- Author
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Apel, Wiebke, Schulze, Waltraud X., and Bock, Ralph
- Subjects
CHLOROPLASTS ,PLANT proteins ,PHOTOSYNTHESIS ,AMINO acids ,AMINOPEPTIDASES ,GENETIC regulation ,PLANT cells & tissues - Abstract
Although chloroplast protein stability has long been recognised as a major level of post-translational regulation in photosynthesis and gene expression, the factors determining protein stability in plastids are largely unknown. Here, we have identified stability determinants in vivo by producing plants with transgenic chloroplasts that express a reporter protein whose N- and C-termini were systematically modified. We found that major stability determinants are located in the N-terminus. Moreover, testing of all 20 amino acids in the position after the initiator methionine revealed strong differences in protein stability and indicated an important role of the penultimate N-terminal amino acid residue in determining the protein half life. We propose that the stability of plastid proteins is largely determined by three factors: (i) the action of methionine aminopeptidase (the enzyme that removes the initiator methionine and exposes the penultimate N-terminal amino acid residue), (ii) an N-end rule-like protein degradation pathway, and (iii) additional sequence determinants in the N-terminal region. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
28. Solar-powered factories for new vaccines and antibiotics
- Author
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Bock, Ralph and Warzecha, Heribert
- Subjects
- *
SOLAR energy industries , *VACCINE biotechnology , *ANTIBIOTICS , *CHLOROPLASTS , *PLANT cell differentiation , *PLASTIDS , *PHOTOSYNTHESIS , *ANTI-infective agents - Abstract
Chloroplasts, the green differentiation form of a group of plant cell organelles called plastids, are the sites of photosynthesis, the main energy source for life on Earth. The small circular genome of the plastid has become increasingly amenable to genetic modification, providing biotechnologists with an attractive site for the accommodation of foreign genes. In recent years, the development of optimized expression strategies has given a huge boost to the exploitation of chloroplasts in molecular farming. Exciting progress has been made with the chloroplast-based production of two particularly important classes of pharmaceuticals: vaccines and antibiotics. Extraordinarily high expression levels and the prospects of developing edible biopharmaceuticals make transgenic chloroplasts a promising platform for the production of next-generation vaccines and antimicrobials. [Copyright &y& Elsevier]
- Published
- 2010
- Full Text
- View/download PDF
29. The paramutated SULFUREA locus of tomato is involved in auxin biosynthesis.
- Author
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Ehlert, Britta, Schöttler, Mark Aurel, Tischendorf, Gilbert, Ludwig-Müller, Jutta, and Bock, Ralph
- Subjects
TOMATO genetics ,AUXIN ,BIOSYNTHESIS ,GENETIC mutation ,PHENOTYPES - Abstract
The tomato (Solanum lycopersicum) sulfurea mutation displays trans-inactivation of wild-type alleles in heterozygous plants, a phenomenon referred to as paramutation. Homozygous mutant plants and paramutated leaf tissue of heterozygous plants show a pigment-deficient phenotype. The molecular basis of this phenotype and the function of the SULFUREA gene (SULF) are unknown. Here, a comprehensive physiological analysis of the sulfurea mutant is reported which suggests a molecular function for the SULFUREA locus. It is found that the sulf mutant is auxin-deficient and that the pigment-deficient phenotype is likely to represent only a secondary consequence of the auxin deficiency. This is most strongly supported by the isolation of a suppressor mutant which shows an auxin overaccumulation phenotype and contains elevated levels of indole-3-acetic acid (IAA). Several lines of evidence point to a role of the SULF gene in tryptophan-independent auxin biosynthesis, a pathway whose biochemistry and enzymology is still completely unknown. Thus, the sulfurea mutant may provide a promising entry point into elucidating the tryptophan-independent pathway of IAA synthesis. [ABSTRACT FROM PUBLISHER]
- Published
- 2008
- Full Text
- View/download PDF
30. Plastid protein synthesis is required for plant development in tobacco.
- Author
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Ahlert, Daniela, Ruf, Stephanie, and Bock, Ralph
- Subjects
CELL metabolism ,CHLOROPLASTS ,GENES ,GENOMES ,PHOTOSYNTHESIS ,PROTEIN synthesis ,CELL lines ,MORPHOLOGY ,CELL division - Abstract
Chloroplasts fulfill important functions in cellular metabolism. The majority of plastid genome-encoded genes is involved in either photosynthesis or chloroplast gene expression. Whether or not plastid genes also can determine extrapiastidic functions has remained controversial. We demonstrate here an essential role of plastid protein synthesis in tobacco leaf development. By using chloroplast transformation, we have developed an experimental system that produces recombination-based knockouts of chloroplast translation in a cell-line-specific manner. The resulting plants are chimeric and, in the presence of translational inhibitors, exhibit severe developmental abnormalities. In the absence of active plastid protein synthesis, leaf blade development is abolished because of an apparent arrest of cell division. This effect appears to be cell-autonomous in that adjacent sectors of cells with translating plastids are phenotypically normal but cannot complement for the absence of plastid translation in mutant sectors. Developmental abnormalities also are seen in flower morphology, indicating that the defects are not caused by inhibited expression of plastid photosynthesis genes. Taken together, our data point to an unexpected essential role of plastid genes and gene expression in plant development and cell division. [ABSTRACT FROM AUTHOR]
- Published
- 2003
- Full Text
- View/download PDF
31. Targeted inactivation of the smallest plastid genome-encoded open reading frame reveals a novel and essential subunit of the cytochrome b6f complex.
- Author
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Hager, Martin, Biehler, Klaus, Illerhaus, Jürgen, Ruf, Stephanie, and Bock, Ralph
- Subjects
AMINO acids ,PLASTIDS ,PEPTIDE hormones ,CHLOROPLASTS ,EXTRACHROMOSOMAL DNA ,PHENOTYPES - Abstract
The smallest conserved open reading frame in the plastid genome, ycf6, potentially specifies a hydrophobic polypeptide of only 29 amino acids. In order to determine the function of this reading frame we have constructed a knockout allele for ycf6. This allele was introduced into the tobacco plastid genome by chloroplast transformation to replace the wild-type ycf6 allele. Homoplasmic Δycf6 plants display a photosynthetically incompetent phenotype. Whereas the two photosystems are intact and physiologically active, we found that the electron transfer from photosystem II to photosystem I is interrupted in Δycf6 plants. Molecular analyses revealed that this block is caused by the complete absence of the cytochrome b
6 f complex, the redox-coupling complex that interconnects the two photosystems. Analysis of purified cytochrome b6 f complex by mass spectroscopy revealed the presence of a protein that has exactly the molecular mass calculated for the Ycf6 protein. This suggests that Ycf6 is a genuine subunit of the cytochrome b6 f complex, which plays a crucial role in complex assembly and/or stability. We therefore propose to rename the ycf6 reading frame petN. [ABSTRACT FROM AUTHOR]- Published
- 1999
- Full Text
- View/download PDF
32. Photosystem I: Its biogenesis and function in higher plants
- Author
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Schöttler, Mark Aurel, Albus, Christin Anne, and Bock, Ralph
- Subjects
- *
PLANT evolution , *PLASTOCYANIN , *FERREDOXIN-NADP reductase , *PHOTOSYNTHESIS , *BIOENERGETICS , *THYLAKOIDS , *MOLECULAR chaperones , *CHLOROPLAST DNA - Abstract
Abstract: Photosystem I (PSI), the plastocyanin-ferredoxin oxidoreductase of the photosynthetic electron transport chain, is one of the largest bioenergetic complexes known. It is composed of subunits encoded in both the chloroplast genome and the nuclear genome and thus, its assembly requires an intricate coordination of gene expression and intensive communication between the two compartments. In this review, we first briefly describe PSI structure and then focus on recent findings on the role of the two small chloroplast genome-encoded subunits PsaI and PsaJ in the stability and function of PSI in higher plants. We then address the sequence of PSI biogenesis, discuss the role of auxiliary proteins involved in cofactor insertion into the PSI apoproteins and in the establishment of protein–protein interactions during subunit assembly. Finally, we consider potential limiting steps of PSI biogenesis, and how they may contribute to the control of PSI accumulation. [Copyright &y& Elsevier]
- Published
- 2011
- Full Text
- View/download PDF
33. Photosynthesis without β-carotene.
- Author
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Pengqi Xu, Chukhutsina, Volha U., Nawrocki, Wojciech J., Schansker, Gert, Bielczynski, Ludwik W., Yinghong Lu, Karcher, Daniel, Bock, Ralph, and Croce, Roberta
- Subjects
- *
CAROTENES , *XANTHOPHYLLS , *ASTAXANTHIN , *PHOTOSYNTHESIS , *ZEAXANTHIN , *CAROTENOIDS , *PHOTOSYSTEMS , *ELECTRON transport - Abstract
Carotenoids are essential in oxygenic photosynthesis: they stabilize the pigment-protein complexes, are active in harvesting sunlight and in photoprotection. In plants, they are present as carotenes and their oxygenated derivatives, xanthophylls. While mutant plants lacking xanthophylls are capable of photoautotrophic growth, no plants without carotenes in their photosystems have been reported so far, which has led to the common opinion that carotenes are essential for photosynthesis. Here, we report the first plant that grows photoautotrophically in the absence of carotenes: a tobacco plant containing only the xanthophyll astaxanthin. Surprisingly, both photosystems are fully functional despite their carotenoid-binding sites being occupied by astaxanthin instead of b-carotene or remaining empty (i.e. are not occupied by carotenoids). These plants display non-photochemical quenching, despite the absence of both zeaxanthin and lutein and show that tobacco can regulate the ratio between the two photosystems in a very large dynamic range to optimize electron transport. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
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