Cyanide-Resistant Respiration in Suspension Cultured Cells of Nicotiana glutinosa L

The respiration of dark-grown Nicotiana glutinosa L. cells in liquid suspension culture was found to be highly cyanide resistant and salicylhydroxamic acid (SHAM) sensitive, indicative of an active alternative respiratory pathway. This was especially true during the lag and logarithmic phases of the 14-day growth cycle. Mitochondria isolated from logarithmically growing cells exhibited active oxidation of malate, succinate, and exogenous NADH. Oxidation of all three substrates had an optimum pH of 6.5 and all were highly resistant to inhibited by cyanide and sensitive to SHAM. Respiratory control was exhibited by all three substrates but only if SHAM was present to block the alternative pathway and divert electrons to the phosphorylating cytochrome pathway. The cyanide-resistant oxidation of exogenous NADH has previously only been associated with Arum spadix mitochondria. Coemergence during evolution of the alternative respiratory pathway and the exogenous NADH dehydrogenase in plant mitochondria as a possible mechanism for removal of cytoplasmic NADH is proposed. Evidence is presented which suggests that mitochondrial assays should be performed at pH 6.5.     

Compartmentation of Alternative Oxidase in Plant Mitochondria

Capacity of alternative pathway mediated, CN-resistant respirationwas measured in mitochondria isolated from three plant species:Iris bulbs, potato tuber callus and Petunia cells, grown insuspension culture. Succinate, NADH or a combination of succinateand NADH were used as respiratory substrates. In all three plantspecies electrons from exogenous NADH appeared to have no accessto the alternative pathway connected with succinate mediatedrespiration and vice versa. Therefore, besides a (functional)compartmentation of Q-pools a compartmentation of alternativepathways in plant mitochondria is proposed as well. (Received October 19, 1985; Accepted January 29, 1986)     

Bio-soliton model that predicts non-thermal electromagnetic frequency bands,that either stabilize or destabilize living cells

Solitons, as self-reinforcing solitary waves, interact with complex biological phenomena such as cellular self-organization. A soliton model is able to describe a spectrum of electromagnetism modalities that can be applied to understand the physical principles of biological effects in living cells, as caused by endogenous and exogenous electromagnetic fields and is compatible with quantum coherence. A bio-soliton model is proposed, that enables to predict which eigen-frequencies of non-thermal electromagnetic waves are life-sustaining and which are, in contrast, detrimental for living cells. The particular effects are exerted by a range of electromagnetic wave eigen-frequencies of one-tenth of a Hertz till Peta Hertz that show a pattern of 12 bands, and can be positioned on an acoustic reference frequency scale. The model was substantiated by a meta-analysis of 240 published articles of biological electromagnetic experiments, in which a spectrum of non-thermal electromagnetic waves were exposed to living cells and intact organisms. These data support the concept of coherent quantized electromagnetic states in living organisms and the theories of Fröhlich, Davydov and Pang. It is envisioned that a rational control of shape by soliton-waves and related to a morphogenetic field and parametric resonance provides positional information and cues to regulate organism-wide systems properties like anatomy, control of reproduction and repair.     

The role of polyamines in animal cell physiology

The ubiquitous distribution of polyamines in nature suggests that they fulfil some fundamental role(s) in living organisms. In animal cells, polyamine content closely parallels changes in the rate of cell proliferation so that the highest content is always observed in rapidly growing cells. The activity of ornithine decarboxylase (which is the first enzyme in the polyamine biosynthetic pathway) has been found to increase significantly in many systems shortly after exposure to hormones. Also, addition of polyamines greatly stimulates cell-free macromolecular synthesis. Observations such as these have suggested that polyamine accumulation stimulates cell growth and is important in the regulation of macromolecular biosynthesis. However, it is also possible to interpret such data as evidence that polyamine accumulation is the result, not the cause, of increased cell growth. This review supports the latter concept and re-examines the significance of the early induction of ornithine decarboxylase activity and of the stimulatory effects of exogenous polyamine on macromolecular synthesis. It is proposed that the polyamines are important only in maintaining cell growth that has already been stimulated by other factors and that their biosynthesis is to a large extent determined by the accumulation of RNA in the cell.     

Neutrophil stimulation and priming by direct contact with activated human T lymphocytes.

The concept that T lymphocytes regulate neutrophil function has an important implication in the understanding of the role of these cells in immunity against infection and in inflammatory diseases, but evidence for this concept is primarily derived from the effects of lymphokines on neutrophils. We now present evidence to show that living or paraformaldehyde-fixed mitogen-activated T lymphocytes, as well as an activated T cell line (HUT-78), induce by cell-cell contact, an oxygen-dependent respiratory burst measured by both the lucigenin-dependent chemiluminescence assay and superoxide production. Neutrophils reacted with purified human T lymphocytes which had been activated by culture in the presence of PHA and PMA for 72 h showed a marked and significant respiratory burst compared with neutrophils treated with T lymphocytes cultured in the absence of these mitogens. Similar results were observed with the paraformaldehyde-fixed T cell line (HUT-78). The ability to stimulate neutrophils required intact paraformaldehyde-fixed T cells, and neutrophil stimulation failed to occur if the T cells and neutrophils were separated by membrane filters. mAb to TNF-alpha, and TNF-beta blocked the ability of rTNF-alpha and TNF-beta to stimulate neutrophils but did not block the neutrophil response induced by activated T cells. Pretreatment of neutrophils with the activated T lymphocytes enhanced the response to the tripeptide, FMLP. It is therefore conceivable that activated T lymphocytes attracted at sites of inflammation influence neutrophil activity by direct plasma membrane interaction which clearly represents an efficient microbial defence mechanism, minimizing tissue damage during inflammation.     

Regulation and Control of Multiple Pathways of Respiratory Metabolism in Relation to Other Physiological Functions in Higher Plants

This account presents the views of the author on the functional and regulatory aspects of respiratory metabolism in higher plants: Control of metabolism (by enzymes) and the interaction of respiration with the other physiological functions in the living plant (metabolic control). This concept, formulated in the early fifties (ref. 47), was presented in part in 1965 (ref. 2) based on experiments performed mostly by the author and his colleagues and by his co-workers in this country. After an interruption of a decade, during which his work was discontinued, a more complete formulation of his views are given here based on results reported by workers in this field in other countries during that period. The more complete view can now be 'summarized as follows: Respiratory metabolism is the process whereby a part of the material stored in the plant (organism) is converted into biological work (function) for maintaining its state of being alive, while the other part of the same material is converted into substances of higher degrees of orderliness (negative entropy) in the form of structure and organization. Within limits imposed by the genetic potential, these processes are controlled by enzymes which in turn are regulated by internal and external factors. The above statement is essentially a special expression of a general view on the functional aspects of living organisms given in the author’s earlier book, Green Thral- dom (Alien & Unwin, London, 1949). If the above theme finds acceptance, it follows, as stated earlier (ref. 14), that: 1. Respiratory metabolic pathways must be multiple ("multilineal") and multi- directional; 2. They must be interacting, not only with themselves, but also with other functions in the plant, alternatingly in time and separately in space (compartmentation); 3. There must be mutual interactions among the pathways and func- tions regulated by enzymes which in turn are regulated through external and internal factors. This functional and regulatory concept of respiratory metabolism in higher plants are now summarized by the following expressions: 1. CH2O + O2→>Xl→X2→H2O + CO2 + E ↓↓ Y1 Y2 in which E = Energy, X1, X2 etc. represent intermediate products, and Y1, Y2 etc. represent anabolic products of different composition and different degrees of complexity. 2. Borrowing from the second law of thermodynamics, the free energy △G deri- ved from process 1 is used for performance of physiological work (function) during which part of the energy is given off in the form of heat (△H), and the other part is concerned with the change of materials of lower orderliness into form and structure with a decrease in entropy (△S): △G = △H - T△S in which T is temperature (in K). This equation may or may not be directly applicable without qualifications in our case. But the decrease in entropy with the change of degree of orderliness in the process of tissue and organ formation from formless materials holds true. 3. The third expression presents the fundamental aspects of our concept of control of metabolism by enzymes and metabolic control of physiological functions. This may be given as: Fuction Gene→ Enzymes→Metabolism→ Structurc State→ Time cource (Solid arrows denote Control) .Experimental evidences selected from the numerous published experimental results, mostly from those of our own, in support of the above scheme at the substrate level oxidation in addition to those given in an earlier account (ref. 2) are presented here. Evidences based on experiments during the past decade on multiple pathways in NADH oxidation through the electron transport chain gathered in the literature (ref. 37) during the period when our work was interrupted completes the formula- tion of our concept on respiratory metabolism at both the substrate and terminal oxidation levels. The use of this generalized concept on the functional and regulatory aspects of respiratory metabolism in higher plants for guiding further research on plant respiration and on other physiological processes, as well as the application of this concept to practical physiological and biological problems are discussed.     

The appearance and characterization of cyanide-resistant respiration in the fungus Candida albicans

The respiration of yeast-form cells of the dimorphic fungus Candida albicans became resistant to cyanide during aging treatment in the resting state. An alternative, cyanide-resistant respiratory pathway was found to develop fully in cells aged at a concentration of 0.75 X 10(9)/ml or more at 25 C, but did not appear at 5 C. Chloramphenicol did not prevent the appearance of the alternative respiratory pathway. The effects of inhibitors, salicylhydroxamic acid (SHAM) and disulfiram (tetraethylthiuram disulfide), on respiration of aged cells were examined, and results indicated that SHAM binds at a site on the alternative respiratory pathway whereas disulfiram binds at two sites, one on the conventional respiratory pathway and the other on the alternative pathway. Thus, SHAM is a more selective inhibitor of the alternative respiration of C. albicans cells. SHAM-titration of the alternative respiration revealed that less than 10% of the maximal activity of the alternative respiratory pathway was utilized under normal conditions, indicating that the alternative respiratory pathway makes a small contribution to the total respiration. It was therefore concluded that the alternative, cyanide-resistant respiratory pathway operates fully when the cyanide-sensitive, cytochrome pathway is blocked although aged cells possess both respiratory pathways.     

Sensitivities of the alternative respiratory components of potato tuber mitochondria to thiol reagents and Ca2+.

Plant mitochondria differ from those of mammals, since they incorporate an alternative electron transport pathway, which branches at ubiquinol to an alternative oxidase (AOX), characteristically inhibited by salicylhydroxamic acid (SHAM). Another feature of plant mitochondria is that besides complex I (EC 1.6.5.3) they possess alternative NAD(P)H-dehydrogenases insensitive to rotenone. Many stress conditions are known to alter the expression of the alternative electron transport pathway in plant mitochondria. In the present study we investigated the effects of some thiol reagents and Ca(2+) on potato mitochondrial respiratory chain presenting different activities of the alternative respiratory components AOX and external NADH dehydrogenase, a condition induced by previous treatment of potato tubers (Solanum tuberosum L., cv. Bintje) to cold stress. The results showed that Ca(2+) presented an inhibitory effect on AOX pathway in potato mitochondria energized with NADH or succinate, which was only now observed when the cytochrome pathway was inhibited by cyanide. When the cytochrome pathway was functional, Ca(2+) stimulated the external NADH dehydrogenase. Diamide was a potent AOX inhibitor and this effect was only now observed when the cytochrome pathway was inactive, as was the case for the calcium ion. Mersalyl inhibited the externally located NADH dehydrogenase and had no effect on AOX activity. The results may represent an important function of Ca(2+) on the alternative mitochondrial enzymes NADH-DH(ext) and AOX.     

Inhibitory effects of nitric oxide on oxidative phosphorylation in plant mitochondria.

Plant nitrate reductase (NR) produces nitric oxide (NO) when nitrite is provided as the substrate in the presence of NADH [H. Yamasaki and Y. Sakihama (2000) FEBS Lett. 468, 89-92]. Using a NR-dependent NO producing system, we investigated the effects of NO on the energy transduction system in plant mitochondria isolated from mung bean (Vigna radiata). Plant mitochondria are known to possess two respiratory electron transport pathways-the cytochrome and alternative pathways. When the alternative pathway was inhibited by n-propyl gallate, the addition of NR strongly suppressed respiratory O(2) consumption driven by the cytochrome pathway. In contrast, the alternative pathway measured in the presence of antimycin A was not affected by NO. The extent of the steady-state membrane potential (Deltapsi) generated by respiratory electron transport rapidly declined in response to NO production. The addition of bovine hemoglobin, a quencher of NO, resulted in the recovery of Deltapsi to the uninhibited level. Consistent with its inhibition of Deltapsi, NO produced by NR strongly suppressed ATP synthesis in the mitochondria. These results provide substantial evidence to confirm that the plant alternative pathway is resistant to NO and support the idea that the alternative pathway may lower respiration-dependent production of active oxygens under conditions where NO is overproduced.     

NaCl胁迫下交替呼吸途径对烟草悬浮细胞死亡调节作用的研究

盐分胁迫是植物在自然环境中经常遭遇的环境胁迫因素之一,会引起植物代谢紊乱乃至细胞死亡,这严重限制了植物的生长、繁育和生存。交替呼吸途径是植物较之动物独特的线粒体呼吸途径。该研究在烟草悬浮细胞中调查了交替呼吸途径对Na Cl胁迫引起的植物细胞死亡过程的调节作用及相应的内在机制,以及在200 mmol·L~(-1)Na Cl处理的烟草悬浮细胞中研究了交替呼吸途径和细胞死亡发生及H_2O_2之间的关系。结果表明:(1)随着Na Cl处理浓度的增加,烟草悬浮细胞死亡水平逐渐增加,而交替呼吸途径的容量也逐渐上升。(2)与Na Cl处理相似,外源H_2O_2的处理也能导致烟草悬浮细胞死亡水平的增加。200 mmol·L~(-1)Na Cl的胁迫导致明显的细胞死亡发生和H_2O_2产量的显著性增加;而较之200 mmol·L~(-1)Na Cl胁迫下的细胞,用水杨基氧肟酸(交替呼吸途径的抑制剂)预处理后的细胞再置于200 mmol·L~(-1)Na Cl的胁迫下导致更高水平的细胞死亡和H_2O_2的产生。综上表明,高盐胁迫诱导了烟草悬浮细胞的交替呼吸途径的增加,而交替呼吸途径则可能通过抑制活性氧的产生而起到缓解细胞死亡发生的作用。     

Origin of the concept of the quiescent centre of plant roots

Concepts in biology feed into general theories of growth, development and evolution of organisms and how they interact with the living and non-living components of their environment. A well-founded concept clarifies unsolved problems and serves as a focus for further research. One such example of a constructive concept in the plant sciences is that of the quiescent centre (QC). In anatomical terms, the QC is an inert group of cells maintained within the apex of plant roots. However, the evidence that established the presence of a QC accumulated only gradually, making use of strands of different types of observations, notably from geometrical-analytical anatomy, radioisotope labelling and autoradiography. In their turn, these strands contributed to other concepts: those of the mitotic cell cycle and of tissue-related cell kinetics. Another important concept to which the QC contributed was that of tissue homeostasis. The general principle of this last-mentioned concept is expressed by the QC in relation to the recovery of root growth following a disturbance to cell proliferation; the resulting activation of the QC provides new cells which not only repair the root meristem but also re-establish a new QC.     

Exponential Signaling Gain at the Receptor Level Enhances Signal-to-Noise Ratio in Bacterial Chemotaxis

Cellular signaling systems show astonishing precision in their response to external stimuli despite strong fluctuations in the molecular components that determine pathway activity. To control the effects of noise on signaling most efficiently, living cells employ compensatory mechanisms that reach from simple negative feedback loops to robustly designed signaling architectures. Here, we report on a novel control mechanism that allows living cells to keep precision in their signaling characteristics – stationary pathway output, response amplitude, and relaxation time – in the presence of strong intracellular perturbations. The concept relies on the surprising fact that for systems showing perfect adaptation an exponential signal amplification at the receptor level suffices to eliminate slowly varying multiplicative noise. To show this mechanism at work in living systems, we quantified the response dynamics of the E. coli chemotaxis network after genetically perturbing the information flux between upstream and downstream signaling components. We give strong evidence that this signaling system results in dynamic invariance of the activated response regulator against multiplicative intracellular noise. We further demonstrate that for environmental conditions, for which precision in chemosensing is crucial, the invariant response behavior results in highest chemotactic efficiency. Our results resolve several puzzling features of the chemotaxis pathway that are widely conserved across prokaryotes but so far could not be attributed any functional role.     

Changes in respiratory mitochondrial machinery and cytochrome and alternative pathway activities in response to energy demand underlie the acclimation of respiration to elevated CO2 in the invasive Opuntia ficus-indica

Studies on long-term effects of plants grown at elevated CO(2) are scarce and mechanisms of such responses are largely unknown. To gain mechanistic understanding on respiratory acclimation to elevated CO(2), the Crassulacean acid metabolism Mediterranean invasive Opuntia ficus-indica Miller was grown at various CO(2) concentrations. Respiration rates, maximum activity of cytochrome c oxidase, and active mitochondrial number consistently decreased in plants grown at elevated CO(2) during the 9 months of the study when compared to ambient plants. Plant growth at elevated CO(2) also reduced cytochrome pathway activity, but increased the activity of the alternative pathway. Despite all these effects seen in plants grown at high CO(2), the specific oxygen uptake rate per unit of active mitochondria was the same for plants grown at ambient and elevated CO(2). Although decreases in photorespiration activity have been pointed out as a factor contributing to the long-term acclimation of plant respiration to growth at elevated CO(2), the homeostatic maintenance of specific respiratory rate per unit of mitochondria in response to high CO(2) suggests that photorespiratory activity may play a small role on the long-term acclimation of respiration to elevated CO(2). However, despite growth enhancement and as a result of the inhibition in cytochrome pathway activity by elevated CO(2), total mitochondrial ATP production was decreased by plant growth at elevated CO(2) when compared to ambient-grown plants. Because plant growth at elevated CO(2) increased biomass but reduced respiratory machinery, activity, and ATP yields while maintaining O(2) consumption rates per unit of mitochondria, we suggest that acclimation to elevated CO(2) results from physiological adjustment of respiration to tissue ATP demand, which may not be entirely driven by nitrogen metabolism as previously suggested.     

一个拖延了半个多世纪的研究课题的总结汇报

本文是汤佩松教授为北京植物生理学会1992年年会准备的发言稿,并以庆祝中国植物生理学会成立30周年。后“北京年会”因故未开,本刊应汤老要求按原样发表于此(并附英文摘要),以飨读者。     

Towards an organismic concept of land plants: The marginal blastozone and the development of the vegetation body of selected frondose gametophytes of liverworts and ferns

An organismic concept of land plants is outlined, which is based on a synthesis of plant morphology and plant anatomy. The entire plant, the living unity, is conceived as the organism being subdivided into cells, which cannot be interpreted as organisms themselves in the sense of elementary organisms. The evolution of land plant tissue systems is discussed in the introductive chapter.To test the proposed concept, some frondose plants were selected from liverworts (Pellia epiphylla, Metzgeria furcata, Pallavicinia lyallii) and comparable fern gametophytes (Dryopteris filix mas, Vittaria lineata, Stenochlaena tenuifolia) and studied with respect to their organization and the principles of development. They all have an archetypic, two-dimensional, open construction, which is described as the repens-type of plant construction. Primary form growth occurs in the marginal blastozone, which controls cell wall integration. One of the most significant processes of form generation is blastozone fractionation. The tissues leaving the blastozone differentiate during extension growth and maturation of the vegetation body. While the plant grows continuously in the blastozone, it decays steadily in the necrozone.The implications of the two-dimensional repens-type are discussed. It appears as a perfect plant construction, fit to start plant evolution on the land surface. Growing upwards into the atmosphere, the repens-type is obscured. But is reappears in all groups of higher land plants. This demonstrates the existence of evolutionary cycles in plants. It is argued that mutation and selection do not suffice to understand cyclical evolutionary patterns. The influence of organismic construction seems to predetermine evolution because of the limited options to change an appropriately functioning construction. Via construction analysis evolutionary options can be detected and thus, evolution becomes predictable to some extent. Instead of being object of mutation and selection, living organisms should be conceived as subjects in evolution (Weingarten 1993).Dedicated to my admired teacher Professor DrWilhelm Troll on the occasion of his 100^th birthday, 3rd November, 1997.     

Pollen profilin function depends on interaction with proline-rich motifs.

The actin binding protein profilin has dramatic effects on actin polymerization in vitro and in living cells. Plants have large multigene families encoding profilins, and many cells or tissues can express multiple profilin isoforms. Recently, we characterized several profilin isoforms from maize pollen for their ability to alter cytoarchitecture when microinjected into living plant cells and for their association with poly-L-proline and monomeric actin from maize pollen. In this study, we characterize a new profilin isoform from maize, which has been designated ZmPRO4, that is expressed predominantly in endosperm but is also found at low levels in all tissues examined, including mature and germinated pollen. The affinity of ZmPRO4 for monomeric actin, which was measured by two independent methods, is similar to that of the three profilin isoforms previously identified in pollen. In contrast, the affinity of ZmPRO4 for poly-L-proline is nearly twofold higher than that of native pollen profilin and the other recombinant profilin isoforms. When ZmPRO4 was microinjected into plant cells, the effect on actin-dependent nuclear position was significantly more rapid than that of another pollen profilin isoform, ZmPRO1. A gain-of-function mutant (ZmPRO1-Y6F) was created and found to enhance poly-L-proline binding activity and to disrupt cytoarchitecture as effectively as ZmPRO4. In this study, we demonstrate that profilin isoforms expressed in a single cell can have different effects on actin in living cells and that the poly-L-proline binding function of profilin may have important consequences for the regulation of actin cytoskeletal dynamics in plant cells.     

Berberine Promotes Glucose Consumption Independently of AMP-Activated Protein Kinase Activation

Berberine is a plant alkaloid with anti-diabetic action. Activation of AMP-activated protein kinase (AMPK) pathway has been proposed as mechanism for berberine’s action. This study aimed to examine whether AMPK activation was necessary for berberine’s glucose-lowering effect. We found that in HepG2 hepatocytes and C2C12 myotubes, berberine significantly increased glucose consumption and lactate release in a dose-dependent manner. AMPK and acetyl coenzyme A synthetase (ACC) phosphorylation were stimulated by 20 µmol/L berberine. Nevertheless, berberine was still effective on stimulating glucose utilization and lactate production, when the AMPK activation was blocked by (1) inhibition of AMPK activity by Compound C, (2) suppression of AMPKα expression by siRNA, and (3) blockade of AMPK pathway by adenoviruses containing dominant-negative forms of AMPKα1/α2. To test the effect of berberine on oxygen consumption, extracellular flux analysis was performed in Seahorse XF24 analyzer. The activity of respiratory chain complex I was almost fully blocked in C2C12 myotubes by berberine. Metformin, as a positive control, showed similar effects as berberine. These results suggest that berberine and metformin promote glucose metabolism by stimulating glycolysis, which probably results from inhibition of mitochondrial respiratory chain complex I, independent of AMPK activation.     

Occurrence, biosynthesis and function of isoprenoid quinones

Isoprenoid quinones are one of the most important groups of compounds occurring in membranes of living organisms. These compounds are composed of a hydrophilic head group and an apolar isoprenoid side chain, giving the molecules a lipid-soluble character. Isoprenoid quinones function mainly as electron and proton carriers in photosynthetic and respiratory electron transport chains and these compounds show also additional functions, such as antioxidant function. Most of naturally occurring isoprenoid quinones belong to naphthoquinones or evolutionary younger benzoquinones. Among benzoquinones, the most widespread and important are ubiquinones and plastoquinones. Menaquinones, belonging to naphthoquinones, function in respiratory and photosynthetic electron transport chains of bacteria. Phylloquinone K₁, a phytyl naphthoquinone, functions in the photosynthetic electron transport in photosystem I. Ubiquinones participate in respiratory chains of eukaryotic mitochondria and some bacteria. Plastoquinones are components of photosynthetic electron transport chains of cyanobacteria and plant chloroplasts. Biosynthetic pathway of isoprenoid quinones has been described, as well as their additional, recently recognized, diverse functions in bacterial, plant and animal metabolism.     

Functioning of a CSP310 stress protein is related to the shunting of electron transfer along the respiratory chain of winter wheat mitochondria

Using three-day-old winter-wheat (Triticum aestivum L.) and six-day-old pea (Pisum sativum L.) seedlings as examples, we studied the effects of inhibitors of the electron transfer chain of plant mitochondria on the uncoupling between oxidation and phosphorylation brought about by the CSP310 stress protein. This uncoupling was inhibited by cyanide and by antibodies against CSP310, but not inhibited by antimycin A. It was shown that, in plant mitochondria, the CSP310 stress protein is involved in the electron transfer via shunting the major cytochrome pathway. In this case, the electron transfer bypasses complex II, ubiquinone, and complex III of the mitochondrial respiratory chain and is realized in the following succession: complex I-CSP310-cytochrome c-complex IV. This electron-transfer pathway was found in winter grass mitochondria during the low-temperature stress and resulted in thermogenesis. It was concluded that CSP310 is a thermogenic system, which is activated in winter grass mitochondria during the low-temperature stress.     

Inhibition of Oxidative Phosphorylation Induces a Rapid Death of GA-Pretreated Aleurone Cells,But Not of ABA-Pretreated Aleurone Cells

Reactive oxygen species (ROS) mediate programmed cell death in aleurone cells, which is promoted by gibberellic acid (GA) and prevented by abscisic acid (ABA). Plant mitochondria contain two distinct respiratory pathways: respiration through cytochrome c oxidase increases ROS production, whereas respiration through the alternative oxidase pathway lowers it. While studying the effects of GA and ABA on partitioning of respiration between those two pathways during the germinating process, we discovered that oxidative phosphorylation inhibitors like sodium azide and 2, 4-dinitrophenol induce rapid death of GA-pretreated aleurone cells but not of ABA-pretreated cells. Functional aerobic respiration was required for GA signaling, and 6 to 12 hours of GA signaling altered the cellular state of aleurone cells to be extremely susceptible to inhibition of oxidative phosphorylation. Anaerobic conditions were also able to mimic the effects of respiratory inhibitors in specifically inducing cell death in GA-treated cells, but cell death was provoked much more slowly. Cotreatment with various antioxidants did not prevent this process at all, suggesting that no ROS are responsible for this respiratory inhibitor-induced cell death. Our observation implicates that GA may partition all the electrons produced during mitochondrial respiration only to the cytochrome oxidase pathway, which would at least partly contribute to cellular accumulation of ROS.