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  • 1. Baba, Kyoko
    et al.
    Schmidt, Julien
    Espinosa-Ruiz, Ana
    Villarejo, Arsenio
    Shiina, Takashi
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Sane, Aniruddha P
    Bhalerao, Rishikesh P
    Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Organellar gene transcription and early seedling development are affected in the rpoT;2 mutant of Arabidopsis.2004In: Plant J, ISSN 0960-7412, Vol. 38, no 1, p. 38-48Article in journal (Refereed)
    Abstract [en]

    An Arabidopsis mutant that exhibited reduced root length was isolated from a population of activation-tagged T-DNA insertion lines in a screen for aberrant root growth. This mutant also exhibited reduced hypocotyl length as well as a delay in greening and altered leaf shape. Molecular genetic analysis of the mutant indicated a single T-DNA insertion in the gene RpoT;2 encoding a homolog of the phage-type RNA polymerase (RNAP), that is targeted to both mitochondria and plastids. A second T-DNA-tagged allele also showed a similar phenotype. The mutation in RpoT;2 affected the light-induced accumulation of several plastid mRNAs and proteins and resulted in a lower photosynthetic efficiency. In contrast to the alterations in the plastid gene expression, no major effect of the rpoT;2 mutation on the accumulation of examined mitochondrial gene transcripts and proteins was observed. The rpoT;2 mutant exhibited tissue-specific alterations in the transcript levels of two other organelle-directed nuclear-encoded RNAPs, RpoT;1 and RpoT;3. This suggests the existence of cross-talk between the regulatory pathways of the three RNAPs through organelle to nucleus communication. These data provide an important information on a role of RpoT;2 in plastid gene expression and early plant development.

  • 2. Bergman, Anders
    et al.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Ericson, Ingemar
    Method to Obtain a Chlorophyll-free Preparation of Intact Mitochondria from Spinach Leaves.1980In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 66, no 3, p. 442-445Article in journal (Refereed)
    Abstract [en]

    Mitochondria from green leaves of spinach have been prepared using a three-step procedure involving differential centrifugation, partition in an aqueous dextran polyethylene glycol two-phase system and Percoll gradient centrifugation. The mitochondrial fractions after the different steps of purification were compared. The final mitochondrial preparation was totally free from chloroplast material measured as chlorophyll content. The enrichment of mitochondria in relation to peroxisomes and microsomes was approximately 12 and 33 times, respectively, based on NAD:isocitrate dehydrogenase activity, glycolate oxidase activity, and NADPH:cytochrome c oxidoreductase activity. The apparent intactness of the inner and the outer mitochondrial membranes was higher than 90% as measured by latency of enzyme activities. The mitochondria showed high respiratory rates with respiratory control and the ADP/O ratios approached the theoretical limits.

  • 3.
    Bhalerao, Rupali
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Keskitalo, Johanna
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Sterky, Fredrik
    Erlandsson, Rikard
    Björkbacka, Harry
    Birve, Simon Jonsson
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Karlsson, Jan
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gustafsson, Petter
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Lundeberg, Joakim
    Jansson, Stefan
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gene expression in autumn leaves2003In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 131, no 2, p. 430-442Article in journal (Refereed)
    Abstract [en]

    Two cDNA libraries were prepared, one from leaves of a field-grown aspen (Populus tremula) tree, harvested just before any visible sign of leaf senescence in the autumn, and one from young but fully expanded leaves of greenhouse-grown aspen (Populus tremula x tremuloides). Expressed sequence tags (ESTs; 5,128 and 4,841, respectively) were obtained from the two libraries. A semiautomatic method of annotation and functional classification of the ESTs, according to a modified Munich Institute of Protein Sequences classification scheme, was developed, utilizing information from three different databases. The patterns of gene expression in the two libraries were strikingly different. In the autumn leaf library, ESTs encoding metallothionein, early light-inducible proteins, and cysteine proteases were most abundant. Clones encoding other proteases and proteins involved in respiration and breakdown of lipids and pigments, as well as stress-related genes, were also well represented. We identified homologs to many known senescence-associated genes, as well as seven different genes encoding cysteine proteases, two encoding aspartic proteases, five encoding metallothioneins, and 35 additional genes that were up-regulated in autumn leaves. We also indirectly estimated the rate of plastid protein synthesis in the autumn leaves to be less that 10% of that in young leaves.

  • 4. BORODIN, V
    et al.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Samuelsson, Göran
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    THE EFFECT OF LIGHT QUALITY ON THE INDUCTION OF EFFICIENT PHOTOSYNTHESIS UNDER LOW CO2 CONDITIONS IN CHLAMYDOMONAS-REINHARDTII AND CHLORELLA-PYRENOIDOSA1994In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 92, no 2, p. 254-260Article in journal (Refereed)
    Abstract [en]

    The effect of blue and red light on the adaptation to low CO2 conditions was studied in high-CO2 grown cultures of Chlorella pyrenoidosa (82T) and Chlamydomonas reinhardtii (137(+)) by measuring O-2 exchange under various inorganic carbon (C-i) concentrations. At equal photosynthetic photon flux density (PPFD), blue light was more favourable for adaptation in both species, compared to red light. The difference in photosynthetic oxygen evolution between cells adapted to low C-i under blue and red light was more pronounced when oxygen evolution was measured under low C-i compared to high C-i conditions. The effect of light quality on adaptation remained for several hours. The different effects caused by blue and red light was observed in C. pyrenoidosa over a wide range of PPFD with increasing differences at increasing PPFD. The maximal difference was obtained at a PPFD above 1 500 mu mol m(-2) s(-1). We found no difference in the extracellular carbonic anhydrase activity between blue- and red light adapted cells. The light quality effect recorded under C-i-limiting conditions in C. reinhardtii cells adapted to air, was only 37% less when instead of pure blue light red light containing 12.5% of blue light (similar PPFD as blue light) was used during adaptation to low carbon. This indicates that in addition to affecting photosynthesis, blue light affected a sensory system involved in algal adaptation to low C-i conditions. Since the affinity for C-i of C. pyrenoidosa and C. reinhardtii cells adapted to air under blue light was higher than that of cells adapted under red light, we suggest that induction of some component(s) of the C-i accumulating mechanism is regulated by the light quality.

  • 5.
    Brouwer, Bastiaan
    et al.
    Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC). Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Keech, Olivier
    Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC). Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    In response to partial plant shading, the lack of phytochrome A does not directly induce leaf senescence but alters the fine-tuning of chlorophyll biosynthesis2014In: Journal of Experimental Botany, ISSN 0022-0957, E-ISSN 1460-2431, Vol. 65, no 14, p. 4037-4049Article in journal (Refereed)
    Abstract [en]

    Phytochrome is thought to control the induction of leaf senescence directly, however, the signalling and molecular mechanisms remain unclear. In the present study, an ecophysiological approach was used to establish a functional connection between phytochrome signalling and the physiological processes underlying the induction of leaf senescence in response to shade. With shade it is important to distinguish between complete and partial shading, during which either the whole or only a part of the plant is shaded, respectively. It is first shown here that, while PHYB is required to maintain chlorophyll content in a completely shaded plant, only PHYA is involved in maintaining the leaf chlorophyll content in response to partial plant shading. Second, it is shown that leaf yellowing associated with strong partial shading in phyA-mutant plants actually correlates to a decreased biosynthesis of chlorophyll rather than to an increase of its degradation. Third, it is shown that the physiological impact of this decreased biosynthesis of chlorophyll in strongly shaded phyA-mutant leaves is accompanied by a decreased capacity to adjust the Light Compensation Point. However, the increased leaf yellowing in phyA-mutant plants is not accompanied by an increase of senescence-specific molecular markers, which argues against a direct role of PHYA in inducing leaf senescence in response to partial shade. In conclusion, it is proposed that PHYA, but not PHYB, is essential for fine-tuning the chlorophyll biosynthetic pathway in response to partial shading. In turn, this mechanism allows the shaded leaf to adjust its photosynthetic machinery to very low irradiances, thus maintaining a positive carbon balance and repressing the induction of leaf senescence, which can occur under prolonged periods of shade.

  • 6.
    Brouwer, Bastiaan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Ziolkowska, Agnieszka
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Bagard, Matthieu
    Keech, Olivier
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    The impact of light intensity on shade-induced leaf senescence2012In: Plant, Cell and Environment, ISSN 0140-7791, E-ISSN 1365-3040, Vol. 35, no 6, p. 1084-1098Article in journal (Refereed)
    Abstract [en]

    Plants often have to cope with altered light conditions, which in leaves induce various physiological responses ranging from photosynthetic acclimation to leaf senescence. However, our knowledge of the regulatory pathways by which shade and darkness induce leaf senescence remains incomplete. To determine to what extent reduced light intensities regulate the induction of leaf senescence, we performed a functional comparison between Arabidopsis leaves subjected to a range of shading treatments. Individually covered leaves, which remained attached to the plant, were compared with respect to chlorophyll, protein, histology, expression of senescence-associated genes, capacity for photosynthesis and respiration, and light compensation point (LCP). Mild shading induced photosynthetic acclimation and resource partitioning, which, together with a decreased respiration, lowered the LCP. Leaf senescence was induced only under strong shade, coinciding with a negative carbon balance and independent of the red/far-red ratio. Interestingly, while senescence was significantly delayed at very low light compared with darkness, phytochrome A mutant plants showed enhanced chlorophyll degradation under all shading treatments except complete darkness. Taken together, our results suggest that the induction of leaf senescence during shading depends on the efficiency of carbon fixation, which in turn appears to be modulated via light receptors such as phytochrome A.

  • 7. Bykova, Natalia V
    et al.
    Keerberg, Olav
    Pärnik, Tiit
    Bauwe, Hermann
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Interaction between photorespiration and respiration in transgenic potato plants with antisense reduction in glycine decarboxylase.2005In: Planta, ISSN 0032-0935, Vol. 222, no 1, p. 130-40Article in journal (Refereed)
    Abstract [en]

    Abstract Potato (Solanum tuberosum L. cv. Désirée) plants with an antisense reduction in the P-protein of the glycine decarboxylase complex (GDC) were used to study the interaction between respiration and photorespiration. Mitochondria isolated from transgenic plants had a decreased capacity for glycine oxidation and glycine accumulated in the leaves. Malate consumption increased in leaves of GDC deficient plants and the capacity for malate and NADH oxidation increased in isolated mitochondria. A lower level of alternative oxidase protein and decreased partitioning of electrons to the alternative pathway was found in these plants. The adenylate status was altered in protoplasts from transgenic plants, most notably the chloroplastic ATP/ADP ratio increased. The lower capacity for photorespiration in leaves of GDC deficient plants was compensated for by increased respiratory decarboxylations in the light. This is interpreted as a decreased light suppression of the tricarboxylic acid cycle in GDC deficient plants in comparison to wild-type plants. The results support the view that respiratory decarboxylations in the light are restricted at the level of the pyruvate dehydrogenase complex and/or isocitrate dehydrogenase and that this effect is likely to be mediated by mitochondrial photorespiratory products.

  • 8. Bykova, Natalia V.
    et al.
    Møller, Ian M.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC). Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Igamberdiev, Abir U.
    The function of glycine decarboxylase complex is optimized to maintain high photorespiratory flux via buffering of its reaction products2014In: Mitochondrion (Amsterdam. Print), ISSN 1567-7249, E-ISSN 1872-8278, Vol. 19, p. 357-364Article in journal (Refereed)
    Abstract [en]

    Oxidation of glycine in photorespiratory pathway is the major flux through mitochondria of C3 plants in the light. It sustains increased intramitochondrial concentrations of NADH and NADPH, which are required to engage the internal rotenone-insensitive NAD(P)H dehydrogenases and the alternative oxidase. We discuss here possible mechanisms of high photorespiratory flux maintenance in mitochondria and suggest that it is fulfilled under conditions where the concentrations of glycine decarboxylase reaction products NADH and CO2 achieve an equilibrium provided by malate dehydrogenase and carbonic anhydrase, respectively. This results in the removal of these products from the glycine decarboxylase multienzyme active sites and in the maintenance of their concentrations at levels sufficiently low to prevent substrate inhibition of the reaction. 

  • 9. Bykova, Natalia V
    et al.
    Rasmusson, Allan G.
    Igamberdiev, Abir U
    Department of Plant Science, Faculty Agriculture and Food Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Møller, Ian M.
    Two separate transhydrogenase activities are present in plant mitochondria1999In: Biochemical and Biophysical Research Communications - BBRC, ISSN 0006-291X, E-ISSN 1090-2104, Vol. 265, no 1, p. 106-111Article in journal (Refereed)
    Abstract [en]

    Inside-out submitochondrial particles from both potato tubers and pea leaves catalyze the transfer of hydride equivalents from NADPH to NAD(+) as monitored with a substrate-regenerating system. The NAD(+) analogue acetylpyridine adenine dinucleotide is also reduced by NADPH and incomplete inhibition by the complex I inhibitor diphenyleneiodonium (DPI) indicates that hive enzymes are involved in this reaction. Gel-filtration chromatography of solubilized mitochondrial membrane complexes confirms that the DPI-sensitive TH activity is due to NADH-ubiquinone oxidoreductase (EC 1,6,5,3, complex I), whereas the DPI-insensitive activity is due to a separate enzyme eluting around 220 kDa. The DPI-insensitive TH activity is specific for the 4B proton on NADH, whereas there is no indication of a 4A-specific activity characteristic of a mammalian-type energy-linked TH. The DPI-insensitive TH may be similar to the soluble type of transhydrogenase found in, e.g., Pseudomonas. The presence of non-energy-linked TR: activities directly coupling the matrix NAD(H) and NADP(H) pools will have important consequences for the regulation of NADP-linked processes in plant mitochondria. (C) 1999 Academic Press.

  • 10. Covey-Crump, Elizabeth M
    et al.
    Bykova, Natalia V
    Affourtit, Charles
    Hoefnagel, Marcel H N
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Atkin, Owen K
    Temperature-dependent changes in respiration rates and redox poise of the ubiquinone pool in protoplasts and isolated mitochondria of potato leaves2007In: Physiologia Plantarum, Vol. 129, p. 175-184Article in journal (Refereed)
    Abstract [en]

    In many environments, leaves experience large diurnal variations in temperature. Such short-term changes in temperature are likely to have important implications for respiratory metabolism in leaves. Here, we used intact leaf, protoplasts and isolated mitochondria to determine the impact of short-term changes in temperature on respiration rates (R), adenylate concentrations and the redox poise of the ubiquinone (UQ) pool in mitochondria of potato leaves. The Q10 (i.e. proportional change in R for each 10°C rise in temperature) of respiration was 1.8, both for intact leaves and protoplasts. In protoplasts, the redox poise of the extracted UQ pool (UQR/UQT) increased from 0.33 at 22°C, to 0.76 at 15°C. Further decreases in temperature (from 15 to 5°C) resulted in UQR/UQT decreasing to 0.40. Adenylate ratios in protoplasts were also temperature dependent. At high adenosine 5'-triphosphate (ATP) adenosine 5'-diphosphate (ADP) ratios (i.e. low ADP concentrations), UQR/UQT values were low, suggesting that adenylates restricted flux via the UQ-reducing pathways more than they restricted flux via pathways that oxidized UQH2. To assess whether high rates of alternative oxidase (AOX) activity could have uncoupled respiratory flux (and thus UQR/UQT) from adenylate restriction of the cytochrome (Cyt) pathway, we constructed kinetic curves of O2 uptake (via the two pathways) vs UQR/UQT in isolated mitochondria, measured at two temperatures (15 and 25°C); measurements were made for mitochondria operating under state 3 (i.e. +ADP) and state 4 (i.e. −ADP) conditions. In contrast to the Cyt pathway, flux via the AOX was temperature insensitive, with maximal rates of AOX activity representing 21–57% of total O2 uptake in isolated mitochondria. We conclude that temperature-dependent variations in UQR/UQT are largely dependent on temperature-dependent changes in adenylate ratios, and that flux via the AOX could in some circumstances help reduce maximal UQ values.

  • 11. EDWARDS, GE
    et al.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    ISOLATION OF MITOCHONDRIA FROM LEAVES OF C-3, C-4, AND CRASSULACEAN ACID METABOLISM PLANTS1987In: Methods in Enzymology, ISSN 0076-6879, E-ISSN 1557-7988, Vol. 148, p. 421-433Article in journal (Refereed)
  • 12. Eriksson, M
    et al.
    Villand, P
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Samuelsson, Göran
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Induction and regulation of expression of a low-CO2-induced mitochondrial carbonic anhydrase in Chlamydomonas reinhardtii1998In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 116, no 2, p. 637-641Article in journal (Refereed)
    Abstract [en]

    The time course of and the influence of light intensity and light quality on the induction of a mitochondrial carbonic anhydrase (CA) in the unicellular green alga Chlamydomonas reinhardtii was characterized using western and northern blots. This CA was expressed only under low-CO2 conditions (ambient air). In asynchronously grown cells, the mRNA was detected 15 min after transfer from air containing 5% CO2 to ambient air, and the 21-kD polypeptide was detected on western blots after 1 h. When transferred back to air containing 5% CO2, the mRNA disappeared within 1 h and the polypeptide was degraded within 3 d. Photosynthesis was required for the induction in asynchronous cultures. The induction increased with light up to 500 mu mol m(-2) s(-1), where saturation occurred. In cells grown synchronously, however, expression of the mitochondrial CA was also detected in darkness. Under such conditions the expression followed a circadian rhythm, with mRNA appearing in the dark 30 min before the light was turned on. Algae left in darkness continued this rhythm for several days.

  • 13. Eriksson, Mats
    et al.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Samuelsson, Göran
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    ISOLATION, PURIFICATION, AND CHARACTERIZATION OF MITOCHONDRIA FROM CHLAMYDOMONAS-REINHARDTII1995In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 107, no 2, p. 479-483Article in journal (Refereed)
    Abstract [en]

    Mitochondria were isolated from autotrophically grown Chlamydomonas reinhardtii cell-wall-less mutant CW 92. The cells were broken by vortexing with glass beads, and the mitochondria were collected by differential centrifugation and purified on a Percoll gradient. The isolated mitochondria oxidized malate, pyruvate, succinate, NADH, and a-ketoglutarate. Respiratory control was obtained with malate (2.0) and pyruvate (2.2) but not with the other substrates. From experiments with KCN and salicylhydroxamic acid, it was estimated that the capacity of the cytochrome pathway was at least 100 nmol O-2 mg(-1) protein min(-1) and the capacity of the alternative oxidase was at least 50 nmol O-2 mg(-1) protein min(-1). A low sensitivity to oligomycin indicates some difference in the properties of the mitochondrial ATPase from Chlamydomonas as compared to higher plants.

  • 14.
    Eriksson, Mats
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Karlsson, Jan
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Ramazanov, Zakir
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Samuelsson, Göran
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Discovery of an algal mitochondrial carbonic anhydrase: molecular cloning and characterization of a low-CO2-induced polypeptide in Chlamydomonas reinhardtii1996In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 93, no 21, p. 12031-12034Article in journal (Refereed)
    Abstract [en]

    In green unicellular algae, several polypeptides are induced upon exposure to limiting CO2. We report here on the localization and characterization of one of these, a 22-kDa polypeptide in Chlamydomonas reinhardtii. This nuclear-encoded polypeptide is induced in the mitochondria by a lowering of the partial pressure of CO2 in the growth medium from 5% to air CO2 levels. Sequencing of two different cDNA clones coding for the polypeptide identified it as a 20.7-kDa beta-type carbonic anhydrase (CA; carbonate dehydratase, carbonate hydro-lyase, EC 4.2.1.1). The two clones differ in their nucleotide sequences but code for identical proteins, showing that this CA is encoded by at least two genes. Northern blot hybridization reveals that mRNA transcripts are only present in cells transferred to air CO2 levels. A comparison of the deduced amino acid sequence with those of other beta-CAs shows the largest degree of similarity with CA from the cyanobacterium Synechocystis (50% identity and 66% similarity). To our knowledge, this is the first identification and characterization of a mitochondrial CA from a photosynthetic organism.

  • 15. Gama, Filipe
    et al.
    Keech, Olivier
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Eymery, Francoise
    Finkemeier, Iris
    Gelhaye, Eric
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Dietz, Karl Josef
    Rey, Pascal
    Jacquot, Jean-Pierre
    Rouhier, Nicolas
    The mitochondrial type II peroxiredoxin from poplar2007In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 129, no 1, p. 196-206Article in journal (Refereed)
    Abstract [en]

    Mitochondria are a major site of reactive oxygen species production and controlling the peroxide levels in this compartment is essential. Peroxiredoxins (Prx) are heme-free peroxidases, which use reactive cysteines for their catalysis and reducing systems for their regeneration. One of the two Prxs present in poplar mitochondria, Prx IIF, expressed as a recombinant protein, was found to reduce a broad range of peroxides with electrons provided preferentially by glutaredoxin and to a lesser extent by glutathione, all the thioredoxins tested being inefficient. This protein is constitutively expressed because it is found in all tissues analyzed. Its expression is modified during a biotic interaction between poplar and the rust fungus Melampsora laricii populina. On the other hand, Prx IIF expression does not substantially vary under abiotic stress conditions. Nevertheless, water deficit or chilling and probably induced senescence, but not photooxidative conditions or heavy metal treatment, also led to a small increase in PrxIIF abundance in Arabidopsis thaliana plants.

  • 16.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    ADENYLATE RATIOS IN THE CYTOSOL, CHLOROPLASTS AND MITOCHONDRIA OF BARLEY LEAF PROTOPLASTS DURING PHOTOSYNTHESIS AT DIFFERENT CARBON-DIOXIDE CONCENTRATIONS1987In: FEBS Letters, ISSN 0014-5793, E-ISSN 1873-3468, Vol. 212, no 1, p. 114-118Article in journal (Refereed)
  • 17.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    METABOLITE LEVELS IN THE CHLOROPLAST AND EXTRACHLOROPLAST COMPARTMENTS OF BARLEY LEAF PROTOPLASTS DURING THE INITIAL PHASE OF PHOTOSYNTHETIC INDUCTION1993In: Biochimica et Biophysica Acta, ISSN 0006-3002, E-ISSN 1878-2434, Vol. 1183, no 2, p. 327-332Article in journal (Refereed)
    Abstract [en]

    Metabolite levels were determined in the chloroplast and extrachloroplast compartments of barley protoplasts during photosynthetic induction using rapid fractionation by membrane filtration. This method allowed studies with a high time resolution the first determination of subcellular metabolite content bring made after only 0.3 s. Upon illumination, dark-adapted protoplasts exhibited a 1 min lag phase prior to commencement of oxygen evolution, and the maximum rate was reached after 4 to 5 min. In contrast to oxygen evolution, the ATP/ADP ratio in the chloroplasts increased from 1 to 2 within 0.5 s and reached a maximum of about 5 after 2 s. There was a dramatic increase in the extrachloroplastic ATP/ADP ratio within a few seconds, reaching a maximum after about 15 s. During the initial phase of photosynthetic induction, the subcellular ATP/ADP ratios were very similar in photorespiratory (low CO,) and non-photorespiratory (high CO,) conditions. The ATP/ADP ratios in both the chloroplast and extrachloroplast compartments remained high until photosynthetic oxygen evolution started and then decreased when the photosynthetic rate reached its maximum. In steady-state photosynthesis the subcellular ATP/ADP ratios were considerably higher under photorespiratory conditions as compared to non-photorespiratory conditions. During the initial phase of photosynthetic induction, 3-phosphoglycerate decreased and triose phosphates increased both in the chloroplast and extrachloroplast compartments. The changes in these metabolites are consistent with a 3-phosphoglycerate/triose phosphate shuttle using the phosphate translocator as the means to supply ATP to the cytosol during photosynthetic induction.

  • 18.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Preparation of leaf mitochondria and studies on mitochondrial photorespiratory reactions1981Doctoral thesis, monograph (Other academic)
    Abstract [en]

    A procedure for the preparation of spinach leaf mitochondria was developed. The procedure combines differential centrifugation, partition in dextran- polyethyleneglycol two-phase system and Percoli density gradient centri- fugation. The different steps separate the material mainly according to size, surface properties and density, respectively. No chlorophyll was present in the final mitochondrial preparation and the mitochondria were also markedly enriched relative to peroxisomes and microsomes as esti­mated from the recovery of marker enzymes. The latency of enzyme activities was used to study the apparent intactness of the mitochondrial membranes. These measurements showed that both the inner and outer mitochondrial membranes were more than 90 % intact. The mitochondria were also functionally intact since the coupling between respiration and oxidative phosphorylation was retained.

    The purity of the preparation made it possible to study cytochromes from leaf mitochondria. The cytochrome content of stalk and leaf mitochondria was measured in order to compare mitochondria from photosynthesizing and non-photosynthesizing tissue. The measurements were performed by difference spectroscopy both at room temperature and at liquid nitrogen temperature. Qualitatively the cytochrome content in mitochondria from stalks and leaves was identical. Quantiatively leaf mitochondria contained,on a protein basis, only half the amount of the different cytochromes as compared to stalk mitochondria. The relative content of the different cytochromes was, however, similar suggesting that the composition of the respiratory chain was the same.

    The photorespiratory conversion of glycine to serine takes place in the mitochondria and involves oxidative decarboxylation of glycine. The ability to oxidize glycine via the respiratory chain was present in spinach leaf mitochondria, but absent in mitochondria prepared from roots, stalks and leaf veins from the same plants. This confirmed the specific localization of the glycine oxidizing activity to photosyntheticaliy active tissue, as suggested by studies with other plant material.

    The conversion of glycine to serine is a complex reaction depending on the combined action of two enzymes: glycine decarboxylase and serine hydroxymethyltransferase. The effect of inhibitors on the serine hydroxy­methyl transferase activity and the rate of the glycine bicarbonate exchange reaction associated with glycine decarboxylase was studied. These reactions represent partial steps in the conversion of glycine to serine and the aim was to investigate the site of inhibition for the different inhibitors, namely, isonicotinyl hydrazide (a pyridoxa!phosphate antagonist), amino- acetonitrile, glycinehydroxamate (glycine analogues) and cyanide. The results showed that these inhibitors had a complex pattern of inhibition. The same inhibitor affected more than one site and often with an apparently different mechanism. It was, however, found that aminoacetonitrile at low concentrations specifically inhibited glycine decarboxylase and that cyanide specifically inhibited serine hydroxymethyltransferase.

  • 19.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Bergman, A
    Ericson, I
    Oxidation of Glycine via the Respiratory Chain in Mitochondria Prepared from Different Parts of Spinach.1980In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 65, no 2, p. 389-91Article in journal (Refereed)
    Abstract [en]

    Mitochondria were prepared from roots, stalks, leaves, and leaf veins of spinach. The mitochondrial preparations were examined for their ability to oxidize glycine via the respiratory chain. It is shown that the glycine-oxidizing capacity is restricted to photosynthetically active tissue. The activity is present in mitochondria from the green parts of the leaves, but not in mitochondria from roots, stalks, or leaf veins.

  • 20.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Edwards, G E
    Isolation of Mitochondria from Leaf Tissue of Panicum miliaceum, a NAD-Malic Enzyme Type C(4) Plant.1983In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 71, no 1, p. 24-9Article in journal (Refereed)
    Abstract [en]

    A mechanical isolation procedure was developed to study the respiratory properties of mitochondria from the mesophyll and bundle sheath tissue of Panicum miliaceum, a NAD-malic enzyme C(4) plant. A mesophyll fraction and a bundle sheath fraction were obtained from young leaves by differential mechanical treatment. The purity of both fractions was about 80%, based on analysis of the cross-contamination of ribulose bisphosphate carboxylase activity and phosphoenolpyruvate carboxylase activity.Mitochondria were isolated from the two fractions by differential centrifugation and Percoll density gradient centrifugation. The enrichment of mitochondria relative to chloroplast material was about 75-fold in both preparations.Both types of mitochondria oxidized NADH and succinate with respiratory control. Malate oxidation in mesophyll mitochondria was sensitive to KCN and showed good respiratory control. In bundle sheath mitochondria, malate oxidation was largely insensitive to KCN and showed no respiratory control. The oxidation was strongly inhibited by salicylhydroxamic acid, showing that the alternative oxidase was involved. The bundle sheath mitochondria of this type of C(4) species contribute to C(4) photosynthesis through decarboxylation of malate. Malate oxidation linked to an uncoupled, alternative pathway may allow decarboxylation to proceed without the restraints which might occur via coupled electron flow through the cytochrome chain.

  • 21.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    EDWARDS, GE
    HENRICSON, D
    ERICSON, I
    THE LOCALIZATION OF SERINE HYDROXYMETHYLTRANSFERASE IN LEAVES OF C-3 AND C-4 SPECIES1985In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 64, no 1, p. 29-33Article in journal (Refereed)
  • 22.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    ERICSON, I
    SEPARATION OF SPINACH LEAF MITOCHONDRIA ACCORDING TO SURFACE-PROPERTIES - PARTITION IN AQUEOUS POLYMER 2-PHASE SYSTEMS1987In: Methods in Enzymology, ISSN 0076-6879, E-ISSN 1557-7988, Vol. 148, p. 434-442Article in journal (Refereed)
  • 23.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Lernmark, U
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    The contribution of mitochondria to energetic metabolism in photosynthetic cells1995In: Journal of Bioenergetics and Biomembranes, ISSN 0145-479X, E-ISSN 1573-6881, Vol. 27, no 4, p. 415-421Article in journal (Refereed)
    Abstract [en]

    Mitochondria fulfill important functions in photosynthetic cells not only in darkness but also in light. Mitochondrial oxidative phosphorylation is probably the main mechanism to supply ATP for extrachloroplastic functions in both conditions. Furthermore, during photosynthesis mitochondrial electron transport is important for regulation of the redox balance in the cell. This makes mitochondrial function an integral part of a flexible metabolic system in the photosynthetic cell. This flexibility is probably very important in order to allow the metabolism to override disturbances caused by the changing environment which plants are adapted to.

  • 24.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Petit, Patrice X
    Møller, Ian M
    Purification and characterization of plant mitochondria and submitochondrial particles1994In: Methods in Enzymology, ISSN 0076-6879, E-ISSN 1557-7988, Vol. 228, p. 424-431Article, review/survey (Refereed)
  • 25.
    Gardeström, Per
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    WIGGE, B
    INFLUENCE OF PHOTORESPIRATION ON ATP/ADP RATIOS IN THE CHLOROPLASTS, MITOCHONDRIA, AND CYTOSOL, STUDIES BY RAPID FRACTIONATION OF BARLEY (HORDEUM-VULGARE) PROTOPLASTS1988In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 88, no 1, p. 69-76Article in journal (Refereed)
  • 26. Goulas, Estelle
    et al.
    Schubert, Maria
    Kieselbach, Thomas
    Umeå University, Faculty of Science and Technology, Department of Chemistry.
    Kleczkowski, Leszek
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Schröder, Wolfgang
    Umeå University, Faculty of Science and Technology, Department of Chemistry. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Hurry, Vaughan
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    The chloroplast lumen and stromal proteomes of Arabidopsis thaliana show differential sensitivity to short- and long-term exposure to low temperature.2006In: Plant Journal, ISSN 0960-7412, Vol. 47, no 5, p. 720-34Article in journal (Refereed)
    Abstract [en]

    Cold acclimation and over-wintering by herbaceous plants are energetically expensive and are dependent on functional plastid metabolism. To understand how the stroma and the lumen proteomes adapt to low temperatures, we have taken a proteomic approach (difference gel electrophoresis) to identify proteins that changed in abundance in Arabidopsis chloroplasts during cold shock (1 day), and short- (10 days) and long-term (40 days) acclimation to 5°C. We show that cold shock (1 day) results in minimal change in the plastid proteomes, while short-term (10 days) acclimation results in major changes in the stromal but few changes in the lumen proteome. Long-term acclimation (40 days) results in modulation of the proteomes of both compartments, with new proteins appearing in the lumen and further modulations in protein abundance occurring in the stroma. We identify 43 differentially displayed proteins that participate in photosynthesis, other plastid metabolic functions, hormone biosynthesis and stress sensing and signal transduction. These findings not only provide new insights into the cold response and acclimation of Arabidopsis, but also demonstrate the importance of studying changes in protein abundance within the relevant cellular compartment.

  • 27. Heineke, D
    et al.
    Bykova, N
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Bauwe, H
    Metabolic response of potato plants to an antisense reduction of the P-protein of glycine decarboxylase2001In: Planta, ISSN 0032-0935, E-ISSN 1432-2048, Vol. 212, no 5-6, p. 880-887Article in journal (Refereed)
    Abstract [en]

    Potato (Solanum tuberosum L. cv. Desire) plants with reduced amounts of P-protein, one of the subunits of glycine decarboxylase (GDC), have been generated by introduction of an antisense transgene. Two transgenic lines, containing about 60-70% less P-protein in the leaves compared to wild-type potato, were analysed in more detail. The reduction in P-protein amount led to a decrease in the ability of leaf mitochondria to decarboxylate glycine. Photosynthetic and growth rates were reduced but the plants were viable under ambient air and produced tubers. Glycine concentrations within the leaves were elevated up to about 100-fold during illumination. Effects: on other amino acids and on sucrose and hexoses were minor. Nearly all of the glycine accumulated during the day was metabolised during the following night. The data suggest that the GDC operates far below substrate saturation under normal conditions thus allowing a flexible and fast response to changes in the environment.

  • 28.
    Hurry, Vaughan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Igamberdiev, Abir U
    Keerberg, Olav
    Pärnik, Tiit
    Atkin, Owen K
    Zaragoza-Costells, Joana
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Respiration in photosynthetic cells2005In: Plant Respiration: From cell to ecosystem, Springer , 2005, p. 43-61Chapter in book (Other academic)
    Abstract [en]

    Respiration in plants, as in all living organisms, is essential to provide metabolic energy and carbon skeletons for growth and maintenance. As such, respiration is an essential component of a plant’s carbon budget. Depending on species and environmental conditions, it consumes 25-75% of all the carbohydrates produced in photosynthesis – even more at extremely slow growth rates. Respiration in plants can also proceed in a manner that produces neither metabolic energy nor carbon skeletons, but heat. This type of respiration involves the cyanide-resistant, alternative oxidase; it is unique to plants, and resides in the mitochondria. The activity of this alternative pathway can be measured based on a difference in fractionation of oxygen isotopes between the cytochrome and the alternative oxidase. Heat production is important in some flowers to attract pollinators; however, the alternative oxidase also plays a major role in leaves and roots of most plants. A common thread throughout this volume is to link respiration, including alternative oxidase activity, to plant functioning in different environments.

  • 29.
    Hurry, Vaughan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Keerberg, O
    Parnik, T
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Öquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Cold-hardening results in increased activity of enzymes involved in carbon metabolism in leaves of winter rye (Secale-Cereale L)1995In: Planta, ISSN 0032-0935, E-ISSN 1432-2048, Vol. 195, no 4, p. 554-562Article in journal (Refereed)
    Abstract [en]

    Light- and CO2-saturated photosynthesis of nonhardened rye (Secale cereale L. cv. Musketeer) was reduced from 18.10 to 7.17 mu mol O-2.m(-2).s(-1) when leaves were transferred from 20 to 5 degrees C for 30 min. Following cold-hardening at 5 degrees C for ten weeks, photosynthesis recovered to 15.05 mu mol O-2.m(-2).s(-1), comparable to the non-hardened rate at 20 degrees C. Recovery of photosynthesis was associated with increases in the total activity and activation of enzymes of the photosynthetic carbon-reduction cycle and of sucrose synthesis. The total hexose-phosphate pool increase by 30% and 120% for nonhardened and cold-hardened leaves respectively when measured at 5 degrees C. The large increase in esterified phosphate in cold-hardened leaves occurred without a limitation in inorganic phosphate supply. In contrast, the much smaller increase in esterified phosphate in nonhardened leaves was associated with an inhibition of ribulose-1,5-bisphosphate carboxylase/oxygenase and sucrose-phosphate synthase activation. It is suggested that the large increases in hexose phosphates in cold-hardened leaves compensates for the higher substrate threshold concentrations needed for enzyme activation at low temperatures. High substrate concentrations could also compensate for the kinetic limitations imposed by product inhibition from the accumulation of sucrose at 5 degrees C. Nonhardened leaves appear to be unable to compensate in this fashion due to an inadequate supply of inorganic phosphate.

  • 30.
    Hurry, Vaughan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Keerberg, O
    Parnik, T
    Öquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Effect of cold hardening on the components of respiratory decarboxylation in the light and in the dark in leaves of winter rye1996In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 111, no 3, p. 713-719Article in journal (Refereed)
    Abstract [en]

    In the dark, all decarboxylation reactions are associated with the oxidase reactions of mitochondrial electron transport. In the light, photorespiration is also active in photosynthetic cells. In winter rye (Secale cereale L.), cold hardening resulted in a P-fold increase in the rate of dark respiratory CO2 release from leaves compared with nonhardened (NH) controls. However, in the light, NH and cold-hardened (CH) leaves had comparable rates of oxidase decarboxylation and total intracellular decarboxylation, Furthermore, whereas CH leaves showed similar rates of total oxidase decarboxylation in the dark and light, NH leaves showed a 2-fold increase in total oxidase activity in the light compared with the dark. Light suppressed oxidase decarboxylation of end products of photosynthesis 2-fold in NH leaves and 3-fold in CH leaves in air. However, in high-CO2, light did not suppress the oxidase decarboxylation of end products. Thus, the decrease in oxidase decarboxylation of end products observed in the light and in air reflected glycolate-cycle-related inhibition of tricarboxylic acid cycle activity. We also showed that the glycolate cycle was involved in the decarboxylation of the end products of photosynthesis in both NH and CH leaves, suggesting a flow of fixed carbon out of the starch pool in the light.

  • 31.
    Hurry, Vaughan M.
    et al.
    Cooperative Research Centre for Plant Science, The Australian National University, Canberra ACT 2601, Australia.
    Strand, Åsa
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Tobiaeson, Maria
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Öquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Cold hardening of spring and winter-wheat and rape results in differential-effects on growth, carbon metabolism, and carbohydrate content1995In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 109, no 2, p. 697-706Article in journal (Refereed)
    Abstract [en]

    The effect of long-term (months) exposure to low temperature (5 degrees C) on growth, photosynthesis, and carbon metabolism was studied in spring and winter cultivars of wheat (Triticum aestivum) and rape (Brassica napus). Cold-grown winter rape and winter wheat maintained higher net assimilation rates and higher in situ CO2 exchange rates than the respective cold-grown spring cultivars. In particular, the relative growth rate of spring rape declined over time at low temperature, and this was associated with a 92% loss in in situ CO2 exchange rates. Associated with the high photosynthetic rates of cold-grown winter cultivars was a P-fold increase per unit of protein in both stromal and cytosolic fructose-1,6-bisphosphatase activity and a 1.5- to 2-fold increase in sucrose-phosphate synthase activity. Neither spring cultivar increased enzyme activity on a per unit of protein basis. We suggest that the recovery of photosynthetic capacity at low temperature and the regulation of enzymatic activity represent acclimation in winter cultivars. This allows these overwintering herbaceous annuals to maximize the production of sugars with possible cryoprotective function and to accumulate sufficient carbohydrate storage reserves to support basal metabolism and regrowth in the spring.

  • 32.
    Hurry, Vaughan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Malmberg, Gunilla
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Öquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Effects of a short-term shift to low-temperature and of long-term cold hardening on photosynthesis and ribulose-1,5-bisphosphate carboxylase oxygenase and sucrose-phosphate synthase activity in leabves of winter rye (Secale-Cereale L)1994In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 106, no 3, p. 983-990Article in journal (Refereed)
    Abstract [en]

    The effect of a short-term (hours) shift to low temperature (5 degrees C) and long-term (months) cold hardening on photosynthesis and carbon metabolism was studied in winter rye (Secale cereale L. cv Musketeer), Cold-hardened plants grown at 5 degrees C exhibited 25% higher in situ CO2 exchange rates than nonhardened plants grown at 24 degrees C. Cold-hardened plants maintained these high rates throughout the day, in contrast to nonhardened plants, which showed a gradual decline in photosynthesis after 3 h. Associated with the increase in photosynthetic capacity following cold hardening was an increase in ribulose-1,5-bisphosphate carboxylase/oxygenase and sucrose phosphate synthase activity and 3- to 4-fold increases in the pools of associated metabolites. Leaves of nonhardened plants shifted overnight to 5 degrees C required 9 h in the light at 5 degrees C before maximum rates of photosynthesis were reached. The gradual increase in photosynthesis in leaves shifted to 5 degrees C was correlated with a sharp decline in the 3-phosphoglycerate/triose phosphate ratio and by an increase in the ribulose bisphosphate/3-phosphoglycerate ratio, indicating the gradual easing of aninorganic phosphate-mediated feedback inhibition on photo-synthesis. We suggest that the strong recovery of photosynthesis in winter rye following cold hardening indicates that the buildup of photosynthetic enzymes, as well as those involved in sucrose synthesis, is an adaptive response that enables these plants to maximize the production of sugars that have both cryoprotective and storage functions that are critical to the performance of these cultivars during over-wintering.

  • 33.
    Hurry, Vaughan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Tobiaeson, M
    Kromer, S
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Öquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Mitochondria contribute to increased photosynthetic capacity of leaves of winter rye (Secale-Cereale L) following cold-hardening1995In: Plant, Cell and Environment, ISSN 0140-7791, E-ISSN 1365-3040, Vol. 18, no 1, p. 69-76Article in journal (Refereed)
    Abstract [en]

    Cold-hardening of winter rye (Secale cereale L. cv. Musketeer) increased dark respiration from -2.2 to -3.9 mu mol O-2 m(-2)s(-1) and doubled light- and CO2-saturated photosynthesis at 20 degrees C from 18.1 to 37.0 mu mol O-2 m(-2) s(-1). We added oligomycin at a concentration that specifically inhibits oxidative phosphorylation to see whether the observed increase in dark respiration reflected an increase in respiration in the light, and whether this contributed to the enhanced photosynthesis of cold-hardened leaves, Oligomycin inhibited light- and CO2-saturated rates of photosynthesis in non-hardened and cold-hardened leaves by 14 and 25%, respectively, and decreased photochemical quenching of chlorophyll a fluorescence to a greater degree in cold-hardened than in non-hardened leaves, These data indicate an increase both in the rate of respiration in the light, and in the importance of respiration to photosynthesis following cold-hardening, Analysis of metabolite pools indicated that oligomycin inhibited photosynthesis by limiting regeneration of ribulose-1,5-bisphosphate, This limitation was particularly severe in cold-hardened leaves, and the resulting low 3-phosphoglycerate pools led to a feed-forward inhibition of sucrose-phosphate synthase activity, Thus, it does not appear that oxidative phosphorylation supports the increase in photosynthetic O-2 evolution following cold-hardening by increasing the availability of cytosolic ATP, The data instead support the hypothesis that the mitochondria function in the light by using the reducing equivalents generated by nan-cyclic photosynthetic electron transport.

  • 34.
    HURRY, VM
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Oquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    REDUCED SENSITIVITY TO PHOTOINHIBITION FOLLOWING FROST-HARDENING OF WINTER RYE IS DUE TO INCREASED PHOSPHATE AVAILABILITY1993In: Planta, ISSN 0032-0935, E-ISSN 1432-2048, Vol. 190, no 4, p. 484-490Article in journal (Refereed)
    Abstract [en]

    The possibility of a role for phosphate metabolism in the photosynthetic regulation that occurs during frost hardening was investigated in winter rye (Secale cereale L. cv. Musketeer). Leaves of frost-hardened and non-hardened winter rye were studied during photosynthetic induction, and at steady state after being allowed to take up 20 mM orthophosphate through the transpiration stream for 3 h. At the growth irradiance (350 mumol.m-2.s-1) frost-hardening increased the stationary rate Of CO2-dependent O2 evolution by 57% and 25% when measured at 5 and 20-degrees-C, respectively. Frost-hardening also reduced the lag phase to stationary photosynthesis by 40% at 5-degrees-C and decreased the susceptibility of leaves to oscillations during induction and after interruption of the actinic beam during steady-state photosynthesis. These responses are all indicative of increased phosphate availability in frost-hardened leaves. As reported previously by Oquist and Huner (1993, Planta 189, 150-156), frost-hardening also decreased the reduction state of Q(A), the primary, stable quinone acceptor of PSII, and decreased the sensitivity of winter rye to photoinhibition of photosynthesis. Non-hardened rye leaves fed orthophosphate also showed an increased photosynthetic capacity (25% at 20-degrees-C and light saturation), lower reduction state of Q(A), a reduced sensitivity to photoinhibition and lower susceptibility to oscillations resulting from a brief interruption of the actinic light. Thus, the data indicate that phosphate metabolism plays a key role in photosynthetic acclimation of winter rye to low temperatures.

  • 35. Igamberdiev, A U
    et al.
    Bykova, N V
    Lea, P J
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    The role of photorespiration in redox and energy balance of photosynthetic plant cells: A study with a barley mutant deficient in glycine decarboxylase2001In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 111, no 4, p. 427-438Article in journal (Refereed)
    Abstract [en]

    Protoplasts and mitochondria were isolated from leaves of homozygous barley (Hordeum vulgare L,) mutant deficient in glycine decarboxylase complex (GDC, EC 2.1.2.10) and wild-type plants. The photosynthetic rates of isolated protoplasts from the mutant and wild-type plants under saturating CO, were similar, but the respiratory rate of the mutant was two-fold higher. Respiration in the mutant plants was much more strongly inhibited by antimycin A than in wild-type plants and a low level of the alternative oxidase protein was found in mitochondria, The activities of NADP- and NAD-dependent malate dehydrogenases were also increased in mutant plants, suggesting an activation of the malate-oxaloacetate exchange for redox transfer between organelles. Mutant plants had elevated activities of NADH- and NADPH-dependent glyoxylate/hydroxypruvate reductases, which may be involved in oxidizing excess NAD(P)H and the scavenging of glyoxylate. We estimated distribution of pools of adenylates, NAD(H) and NADP(H) between chloroplasts, cytosol and mitochondria. Under photorespiratory conditions, ATP/ADP and NADPH/NADP ratios in the mutant were higher in chloroplasts as compared to wild-type plants. The cytosolic NADH/NAD ratio was increased, whereas the ratio in mitochondria decreased. It is concluded that photorespiration serves as an effective redox transfer mechanism from the chloroplast, Plants with a lowered GDC content are deficient in this mechanism, which leads to over-reduction and over-energization of the chloroplasts.

  • 36. Igamberdiev, A U
    et al.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Regulation of NAD- and NADP-dependent isocitrate dehydrogenases by reduction levels of pyridine nucleotides in mitochondria and cytosol of pea leaves2003In: Biochimica et Biophysica Acta - Bioenergetics, ISSN 0005-2728, E-ISSN 1879-2650, Vol. 1606, no 1-3, p. 117-125Article in journal (Refereed)
    Abstract [en]

    Regulation of NAD- and NADP-dependent isocitrate dehydrogenases (NAD-ICDH, EC 1.1.1.41, and NADP-ICDH, EC 1.1.1.42) by the level of reduced and oxidized pyridine nucleotides has been investigated in pea (Pisum sativum L.) leaves. The affinities of mitochondrial and cytosolic ICDH enzymes to substrates and inhibitors were determined on partially purified preparations in forward and reverse directions. From the kinetic data, it follows that NADP(+)- and NAD+-dependent isocitrate dehydrogenases in mitochondria represent a system strongly responding to the intramitochondrial NADPH and NADH levels. The NADPH, NADP(+), NADH and NAD(+) concentrations were determined by subcellular fractionation of pea leaf protoplasts using membrane filtration in mitochondria and cytosol in darkness and in the light under saturating and limiting CO2 Conditions. The cytosolic NADPH/NADP ratio was about I and almost constant both in darkness and in the light. In mitochondria, the NADPH/NADP ratio was low in darkness (0.2) and increased in the light, reaching 3 in limiting CO2 conditions compared to I in saturating CO2. At high reduction levels of NADP and NAD observed at limiting CO2 in the light, i.e. when photorespiratory glycine is the main mitochondrial substrate, isocitrate oxidation in mitochondria will be suppressed and citrate will be transported to the cytosol ('citrate valve'), where the cytosolic NADP-ICDH supplies 2-oxoglutarate for the photorespiratory ammonia refixation. (C) 2003 Elsevier B.V. All rights reserved.

  • 37. Igamberdiev, A U
    et al.
    Ivlev, A A
    Bykova, N V
    Threlkeld, C N
    Lea, P J
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Decarboxylation of glycine contributes to carbon isotope fractionation in photosynthetic organisms2001In: Photosynthesis Research, ISSN 0166-8595, E-ISSN 1573-5079, Vol. 67, no 3, p. 177-184Article in journal (Refereed)
    Abstract [en]

    Carbon isotope effects were investigated for the reaction catalyzed by the glycine decarboxylase complex (GDC; EC 2.1.2.10). Mitochondria isolated from leaves of pea (Pisum sativum L.) and spinach (Spinacia oleracea L.) were incubated with glycine, and the CO2 evolved was analyzed for the carbon isotope ratio (delta C-13). Within the range of parameters tested (temperature, pH, combination of cofactors NAD(+), ADP, pyridoxal 5-phosphate), carbon isotope shifts of CO2 relative to the C-1-carboxyl carbon of glycine varied from +14 parts per thousand to -7 parts per thousand. The maximum effect of cofactors was observed for NAD(+), the removal of which resulted in a strong C-12 enrichment of the CO2 evolved. This indicates the possibility of isotope effects with both positive and negative signs in the GDC reaction. The measurement of delta C-13 in the leaves of the GDC-deficient barley ( Hordeum vulgare L.) mutant (LaPr 87/30) plants indicated that photorespiratory carbon isotope fractionation, opposite in sign when compared to the carbon isotope effect during CO2 photoassimilation, takes place in vivo. Thus the key reaction of photorespiration catalyzed by GDC, together with the key reaction of CO2 fixation catalyzed by ribulose-1,5-bisphosphate carboxylase, both contribute to carbon isotope fractionation in photosynthesis.

  • 38. Igamberdiev, A U
    et al.
    Romanowska, E
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Photorespiratory flux and mitochondrial contribution to energy and redox balance of barley leaf protoplasts in the light and during light-dark transitions2001In: Journal of plant physiology (Print), ISSN 0176-1617, E-ISSN 1618-1328, Vol. 158, no 10, p. 1325-1332Article in journal (Refereed)
    Abstract [en]

    The contribution of mitochondrial oxidation of photorespiratory and respiratory substrates to subcellular energy and redox balance was investigated in leaf protoplasts of barley (Hordeum vulgare L.). The ATP/ADP ratios (indicating the energy balance) in chloroplasts and in tine extrachloroplast compartment were highest in the light in limiting CO2 (photorespiratory conditions), and they drastically increased after illumination if plants were pre-incubated in darkness for 24 hours. After illumination, the ATP/ADP ratio rapidly decreased in chloroplasts. The NADPH/NADP ratio (as an indicator of redox balance) in chloroplasts declined rapidly during the first seconds of darkness, then slowly increased. In limiting CO2, the ratio decreased more slowly during the first minute of darkness corresponding to post-illumination respiratory burst (PIE). During this period, the activation state of chloroplast NADP-malate dehydrogenase was higher in limiting CO2 than in saturating CO2. However, during the light-enhanced dark respiration (LEDR) period, following PIE, there were no differences in subcellular NADPH/NADP ratios in saturating and limiting CO2. A decline in malate and citrate concentrations in protoplasts and activation of mitochondrial NAD-malic enzyme were revealed during LEDR. The results presented highlight the importance of glycine oxidation in mitochondria in energization of the cytosol and chloroplasts and in maintaining redox balance in the light and during the first minute after illumination. And further, they show non-photorespiratory origin of LEDR.

  • 39.
    Igamberdiev, Abir U
    et al.
    Department of Plant Physiology and Biochemistry, Voronezh University, Voronezh 394693, Russia.
    Bykova, Natalia V
    Department of Plant Physiology and Biochemistry, Voronezh University, Voronezh 394693, Russia.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Involvement of cyanide-resistant and rotenone-insensitive pathways of mitochondrial electron transport during oxidation of glycine in higher plants1997In: FEBS Letters, ISSN 0014-5793, E-ISSN 1873-3468, Vol. 412, no 2, p. 265-269Article in journal (Refereed)
    Abstract [en]

    Metabolism of glycine in isolated mitochondria and protoplasts was investigated in photosynthetic, etiolated (barley and pea leaves) and fat-storing (maize scutellum) tissues using methods of [1-C-14]glycine incorporation and counting of (CO2)-C-14 evolved, oxymetric measurement of glycine oxidation and rapid fractionation of protoplasts incubated in photorespiratory conditions with consequent determination of ATP/ADP ratios in different cell compartments, The involvement of different paths of electron transport in mitochondria during operation of glycine decarboxylase complex (GDC) was tested in different conditions, using aminoacetonitrile (AAN), the inhibitor of glycine oxidation in mitochondria, rotenone, the inhibitor of Complex I of mitochondrial electron transport, and inhibitors of cytochrome oxidase and alternative oxidase, It was shown that glycine has a preference to other substrates oxidized in mitochondria only in photosynthetic tissue where succinate and malate even stimulated its oxidation, Rotenone had no or small effect on glycine oxidation, whereas the role of cyanide-resistant path increased in the presence of ATP, Glycine oxidation increased ATP/ADP ratio in cytosol of barley protoplasts incubated in the presence of CO2, but not in the CO2-free medium indicating that in conditions of high photorespiratory nus oxidation of NADH formed in the GDC reaction passes via the non-coupled paths, Activity of GDC in fat-storing tissue correlated with the activity of glyoxylate-cycle enzymes, glycine oxidation did not reveal preference to other substrates and the involvement of paths non-connected with proton translocation was not pronounced, It is suggested that the preference of glycine to other substrates oxidized in mitochondria is achieved in photosynthetic tissue by switching to rotenone-insensitive intramitochrondrial NADH oxidation and by increasing of alternative oxidase involvement in the presence of glycine. (C) 1997 Federation of European Biochemical Societies.

  • 40.
    Igamberdiev, Abir U
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Hurry, Vaughan
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Krömer, Silke
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    The role of mitochondrial electron transport during photosynthetic induction. A study with barley (Hordeum vulgare) protoplasts incubated with rotenone and oligomycin1998In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 104, no 3, p. 431-439Article in journal (Refereed)
    Abstract [en]

    Mitochondrial contribution to photosynthetic metabolism during photosynthetic induction was investigated in protoplasts from barley leaves (Hordeum vulgare L. cv. Gunilla, Svalof) by using an inhibitor of mitochondrial Complex I (rotenone) and an inhibitor of the mitochondrial ATPase (oligomycin). Both inhibitors increased the lag phase of photosynthetic induction after the transition of protoplasts from darkness to light. This effect was not observed with broken protoplasts or isolated chloroplasts. Using the method of rapid fractionation of protoplasts it was shown that the delay in photosynthetic induction was accompanied by a decrease in ATP/ADP ratios of the cytosol and mitochondria, whereas the ratio in chloroplasts was not affected. A delay in activation of chloroplastidic NADP-dependent malate dehydrogenase (EC 1.1.1.82) was observed in the presence of either inhibitor. A delay was also observed in the rise of photochemical quenching of chlorophyll fluorescence in the presence of rotenone or oligomycin during photosynthetic induction. The results indicate that during the transition from dark to light the mitochondrial electron transport chain and its Complex I participate in the reoxidation of excessive redox equivalents from photosynthetic electron transport.

  • 41. Igamberdiev, Abir U.
    et al.
    Lernmark, Ulrika
    Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC). Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Activity of the mitochondrial pyruvate dehydrogenase complex in plants is stimulated in the presence of malate2014In: Mitochondrion (Amsterdam. Print), ISSN 1567-7249, E-ISSN 1872-8278, Vol. 19, no Part B, p. 184-190Article in journal (Refereed)
    Abstract [en]

    The effect of malate on the steady-state activity of the pea (Pisum sativum L.) and barley (Hordeum vulgare L) leaf pyruvate dehydrogenase complex (PDC) has been studied in isolated mitochondria. The addition of malate was found to be stimulatory for the mitochondrial PDC, however there was no stimulation of chloroplast PDC. The stimulation was saturated below 1 mM malate and was apparently related to a partially activated complex, which activity increased in the presence of malate by about twofold. Malate also reversed the reduction of PDC activity in the presence of glycine. Based on the obtained kinetic data, we suggest that the effect of malate is rather not a direct activation of PDC but involves the establishment of NAD-malate dehydrogenase equilibrium, decreasing concentration of NADH and relieving its inhibitory effect of PDC. 

  • 42. Igamberdiev, Abir U
    et al.
    Mikkelsen, Teis N
    Ambus, Per
    Bauwe, Herrmann
    Lea, Peter J
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Photorespiration Contributes to Stomatal Regulation and Carbon Isotope Fractionation: A Study with Barley, Potato and Arabidopsis Plants Deficient in Glycine Decarboxylase2004In: Photosynthesis Research, Vol. 81, p. 139-152Article in journal (Refereed)
    Abstract [en]

    The rates of respiration in light and darkness, C i/C a and carbon isotope fractionation were investigated in glycine decarboxylase-deficient plants of barley, potato and Arabidopsis thaliana grown in climate chambers with controlled light intensity, temperature, humidity, irradiation and different CO2 concentrations (360, 700 and 1400 µl l–1) and compared to the wild-type plants. All photorespiration-impaired plants exhibited higher C i/C a and corresponding lower apparent water-use efficiencies, which were more expressed under high irradiance and elevated temperature. The mutants were depleted in 13C as compared to the wild-type plants, with a difference of up to 6permil following growth in 360 µl l–1 CO2. We determined the carbon isotope content at different CO2 concentrations to calculate the contribution of both C i/C a and photorespiration for 13C/12C fractionation. The direct effect of photorespiration was in the range of 0.7–1.0permil, from which we calculated the value of fractionation at the site of glycine decarboxylation as being 10–13permil, which is in agreement with the previously reported carbon isotope discrimination exerted by the glycine decarboxylase. Respiratory rates, particularly in the light, were increased in the glycine decarboxylase mutants. The necessity of the maintenance of a high CO2 concentration near the site of carboxylation in chloroplasts in plants deficient in photorespiratory enzymes, requires an increased opening of the stomata with a corresponding decrease in water-use efficiency. It is concluded that photorespiration participates in the regulation of C i/C a and contributes to carbon isotope fractionation, both via effects on stomata and via discrimination of 13C in the glycine decarboxylase reaction.

  • 43. Igamberdiev, Abir U
    et al.
    Shen, Tongyun
    Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Function of mitochondria during the transition of barley protoplasts from low light to high light.2006In: Planta, ISSN 0032-0935, Vol. 224, no 1, p. 196-204Article in journal (Refereed)
    Abstract [en]

    Mitochondrial contribution to photosynthetic metabolism during the transition from low light (25–100 μmol quanta m−2 s−1, limiting photosynthesis) to high light (500 μmol quanta m−2 s−1, saturating photosynthesis) was investigated in protoplasts from barley (Hordeum vulgare) leaves. After the light shift, photosynthetic oxygen evolution rate increased rapidly during the first 30–40 s and then declined up to 60–70 s after which the rate increased to a new steady-state after 80–110 s. Rapid fractionation of protoplasts was used to follow changes in sub-cellular distribution of key metabolites during the light shift and the activation state of chloroplastic NADP-dependent malate dehydrogenase (EC 1.1.1.82) was measured. Although oligomycin (an inhibitor of the mitochondrial ATP synthase) affected the metabolite content of protoplasts following the light shift, the first oxygen burst was not affected. However, the transition to the new steady-state was delayed. Rotenone (an inhibitor of mitochondrial complex I) had similar, but less pronounced effect as oligomycin. From the analysis of metabolite content and sub-cellular distribution we suggest that the decrease in oxygen evolution following the first oxygen burst is due to phosphate limitation in the chloroplast stroma. For the recovery the control protoplasts can utilize ATP supplied by mitochondrial oxidative phosphorylation to quickly overcome the limitation in stromal phosphate and to increase the content of Calvin cycle metabolites. The oligomycin-treated protoplasts were deficient in cytosolic ATP and thereby unable to support Calvin cycle operation. This resulted in a delayed capacity to adjust to a sudden increase in light intensity.

  • 44.
    Igamberdiev, Abir U
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Zhou, Guoquing
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Malmberg, Gunilla
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Respiration of barley protoplasts before and after illumination1997In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 99, no 1, p. 15-22Article in journal (Refereed)
    Abstract [en]

    Respiratory O-2 consumption was investigated in dark-adapted barley (Hordeum vulgare L. cv. Gunilla) protoplasts and after illumination for 10 min at high and very low CO2 in the presence of respiratory and photorespiratory inhibitors. In dark-adapted protoplasts no difference was observed between inhibitor treatments in high and very low CO2. The respiratory rate increased somewhat after illumination and a difference in responce to inhibitors was in some cases observed between high and very low CO2. Thus, the operation of the mitochondrial electron transport chain is affected following a period of active photosynthesis. In all situations tested, oligomycin inhibited respiratiory O-2 uptake indicating that respiration of mitochondria in protoplasts is not strictly ADP limited. Antimycin A inhibited respiration more in dark-adapted protoplasts than after illumination whereas SHAM gave the opposite response. Rotenone inhibited respiration both in dark adapted protoplasts (about 30%) and after illumination where the inhibition was much greater in very low CO2 (50%) than in high CO2 (10%). After iilumination in very low CO2, SHAM + rotenone inhibited respiration almost completely (70%). Photorespiratory inhibitors had very small effect on O-2 consumption in darkness. After illumination the effect of aminoacetonitrile (AAN) was also very low whereas a-hydroxypyridine-2-methane sulphonate (HPMS) in photorespiratory conditions inhibited O-2 uptake much stronger (35%). The addition of glyoxylate enhanced respiration in the presence of HPMS up to the control level suggesting that alternative pathways of glyoxylate conversion might be operating. The differences in inhibitor responses may reflect fine mechanisms for the regulation of energetic balance in the plant cell which consists of switching from electron transport coupled to ATP production to non-coupled transport. Photorespiratory flux is also very flexible, and the suppression of glycine decarboxylation can induce bypass reactions of glyoxylate metabolism.

  • 45. Ivanov, A G
    et al.
    Sane, P V
    Zeinalov, Y
    Malmberg, G
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Huner, N P A
    Oquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Photosynthetic electron transport adjustments in overwintering Scots pine (Pinus sylvestris L.)2001In: Planta, ISSN 0032-0935, E-ISSN 1432-2048, Vol. 213, no 4, p. 575-585Article in journal (Refereed)
    Abstract [en]

    As shown before [C. Ottander et al. (1995) Planta 197:176-183], there is a severe inhibition of the photosystem (PS) II photochemical efficiency of Scots pine (Pinus sylvestris L.) during the winter. In contrast, the in vivo PSI photochemistry is less inhibited during winter as shown by in vivo measurements of DeltaA(820)/Delta (820) (P700(+)). There was also an enhanced cyclic electron transfer around PSI in winter-stressed needles as indicated by 4-fold faster reduction kinetics of P700(+). The differential functional stability of PSII and PSI was accompanied by a 3.7-fold higher intersystem electron pool size, and a 5-fold increase in the stromal electron pool available for P700(+) reduction. There was also a strong reduction of the QB band in the thermoluminescence glow curve and markedly slower Q-A re-oxidation in needles of winter pine, indicating an inhibition of electron transfer between QA and QB. The data presented indicate that the plastoquinone pool is largely reduced in winter pine, and that this reduced state is likely to be of metabolic rather than photochemical origin. The retention of PSI photochemistry, and the suggested metabolic reduction of the plastoquinone pool in winter stressed needles of Scots pine are discussed in terms of the need for enhanced photoprotection of the needles during the winter and the role of metabolically supplied energy for the recovery of photosynthesis from winter stress in evergreens.

  • 46. Ivanov, Alexander G
    et al.
    Krol, Marianna
    Sveshnikov, Dimitri
    Malmberg, Gunilla
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Hurry, Vaughan
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Öquist, Gunnar
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Huner, Norman P A
    Characterization of the photosynthetic apparatus in cortical bark chlorenchyma of Scots pine.2006In: Planta, ISSN 0032-0935, Vol. 223, no 6, p. 1165-77Article in journal (Refereed)
    Abstract [en]

    Winter-induced inhibition of photosynthesis in Scots pine (Pinus sylvestris L.) needles is accompanied by a 65% reduction of the maximum photochemical efficiency of photosystem II (PSII), measured as F v/F m, but relatively stable photosystem I (PSI) activity. In contrast, the photochemical efficiency of PSII in bark chlorenchyma of Scots pine twigs was shown to be well preserved, while PSI capacity was severely decreased. Low-temperature (77 K) chlorophyll fluorescence measurements also revealed lower relative fluorescence intensity emitted from PSI in bark chlorenchyma compared to needles regardless of the growing season. Nondenaturating SDS-PAGE analysis of the chlorophyll–protein complexes also revealed much lower abundance of LHCI and the CPI band related to light harvesting and the core complex of PSI, respectively, in bark chlorenchyma. These changes were associated with a 38% reduction in the total amount of chlorophyll in the bark chlorenchyma relative to winter needles, but the Chl a/b ratio and carotenoid composition were similar in the two tissues. As distinct from winter pine needles exhibiting ATP/ADP ratio of 11.3, the total adenylate content in winter bark chlorenchyma was 2.5-fold higher and the estimated ATP/ADP ratio was 20.7. The photochemical efficiency of PSII in needles attached to the twig recovered significantly faster (28–30 h) then in detached needles. Fluorescence quenching analysis revealed a high reduction state of Q A and the PQ-pool in the green bark tissue. The role of bark chlorenchyma and its photochemical performance during the recovery of photosynthesis from winter stress in Scots pine is discussed.

  • 47.
    Karlsson, Jan
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    RAMAZANOV, Z
    Hiltonen, Thomas
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Samuelsson, Göran
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    EFFECT OF VANADATE ON PHOTOSYNTHESIS AND THE ATP ADP RATIO IN LOW-CO2-ADAPTED CHLAMYDOMONAS-REINHARDTII CELLS1994In: Planta, ISSN 0032-0935, E-ISSN 1432-2048, Vol. 192, no 1, p. 46-51Article in journal (Refereed)
    Abstract [en]

    We have assessed the effect of vanadate as an inhibitor of plasma-membrane ATPase on photosynthesis and the ATP/ADP ratio in Chlamydomonas reinhardtii CW-92 (a mutant strain lacking a cell wall). This effect was compared in low-CO2-adapted cells grown in media bubbled with air containing 400 or 70 muL . L-1 CO2. Evidence is presented indicating that cells grown at 70 muL . L-1 CO2 have a higher rate of photosynthetic O2 evolution than cells grown at 400 muL . L-1 CO2, at limiting carbon concentrations. Extracellular and intracellular carbonic-anhydrase activities were, however, similar in cells grown in both of the low-carbon conditions. Vanadate inhibited, to a different extent, the HCO3--dependent O2 evolution in cells grown at 400 and 70 muL . L-1 CO2. At 400 muM vanadate, inhibition reached 70-75 % in cells grown at 400 muL . L-1 but only 50 % in those grown at 70 muL . L-1 CO2. The ATP/ADP ratios determined with and without vanadate at limiting concentrations of dissolved inorganic carbon indicated that more ATP was hydrolysed in algae grown at 70 muL . L-1 than in those grown at 400 muL . L-1 CO2. We conclude that the maximal capacity to accumulate dissolved inorganic carbon is inversely related to the CO2 concentration in the medium. Activation and - or synthesis of vanadate-sensitive ATPase may be the major explanation for the higher capacity for HCO3--dependent O2 evolution in cells grown under limited CO2 concentrations.

  • 48. KAY, CJ
    et al.
    ERICSON, I
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    PALMER, JM
    MOLLER, IM
    GENERATION AND PURIFICATION OF SUBMITOCHONDRIAL PARTICLES OF DIFFERENT POLARITIES FROM PLANT-MITOCHONDRIA1985In: FEBS Letters, ISSN 0014-5793, E-ISSN 1873-3468, Vol. 193, no 2, p. 169-174Article in journal (Refereed)
  • 49.
    Keech, Olivier
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Dizengremel, Pierre
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Preparation of leaf mitochondria from Arabidopsis thaliana2005In: Physiologia Plantarum: An International Journal for Plant Biology, ISSN 0031-9317, E-ISSN 1399-3054, Vol. 124, no 4, p. 403-409Article in journal (Refereed)
    Abstract [en]

    Arabidopsis thaliana is, perhaps, the most important model species in modern plant biology. However, the isolation of organelles from leaves of this plant has been difficult. Here, we present two different protocols for the isolation of mitochondria, yielding either highly functional crude mitochondria or highly purified mitochondria. The crude mitochondria were well coupled with the substrates tested (malate + glutamate, glycine and NADH), exhibiting respiratory control ratios of 2.1–3.9. Purified mitochondria with very low levels of chlorophyll contamination were obtained by Percoll gradient centrifugation, yielding 1.2 mg of mitochondrial protein from 50 g of leaves.

  • 50.
    Keech, Olivier
    et al.
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Pesquet, Edouard
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Gutierrez, Laurent
    Ahad, Abdul
    Bellini, Catherine
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Smith, Steven M
    Gardeström, Per
    Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
    Leaf senescence is accompanied by an early disruption of the microtubule network in Arabidopsis.2010In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 154, no 4, p. 1710-1720Article in journal (Refereed)
    Abstract [en]

    The dynamic assembly and disassembly of microtubules (MTs) is essential for cell function. Although leaf senescence is a well-documented process, the role of the MT cytoskeleton during senescence in plants remains unknown. Here, we show that both natural leaf senescence and senescence of individually darkened Arabidopsis (Arabidopsis thaliana) leaves are accompanied by early degradation of the MT network in epidermis and mesophyll cells, whereas guard cells, which do not senesce, retain their MT network. Similarly, entirely darkened plants, which do not senesce, retain their MT network. While genes encoding the tubulin subunits and the bundling/stabilizing MT-associated proteins (MAPs) MAP65 and MAP70-1 were repressed in both natural senescence and dark-induced senescence, we found strong induction of the gene encoding the MT-destabilizing protein MAP18. However, induction of MAP18 gene expression was also observed in leaves from entirely darkened plants, showing that its expression is not sufficient to induce MT disassembly and is more likely to be part of a Ca(2+)-dependent signaling mechanism. Similarly, genes encoding the MT-severing protein katanin p60 and two of the four putative regulatory katanin p80s were repressed in the dark, but their expression did not correlate with degradation of the MT network during leaf senescence. Taken together, these results highlight the earliness of the degradation of the cortical MT array during leaf senescence and lead us to propose a model in which suppression of tubulin and MAP genes together with induction of MAP18 play key roles in MT disassembly during senescence.

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