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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 49 (1980), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Ethanol, when added to the incubation medium of callus-forming potato tuber discs, inhibits callus growth and causes an increase of the mitochondrial antimycin-A resistant respiration, expressed as a percentage of state III-respiration. This increase in resistance to antimycin-A is the result of a poor development of the cytochrome pathway in tissue discs treated with ethanol. The development of the antimycin-A resistant alternative oxidase sensitive to chelator is about the same for treated and untreated discs.The respiratory control (RC) ratio of the mitochondrial respiration increases after addition of a chelator, which inhibits the alternative pathway. The RC ratio of the uninhibited mitochondrial respiration appears to be inversely related to the capacity of the alternative pathway, when mitochondrial preparations with different capacities to transfer electrons via the alternative path are compared. From the experimentally observed relation between RC-ratio and alternative oxidase capacity, it was concluded that at least half of the capacity of the alternative path is used in uninhibited state IV respiration.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: In potato (Solatium tuberosum L. cv. Bintje and Doré) callus a very active hydrox-amate-stimulated NADH-dependent O2-uptake develops during the growth of the callus, which is caused by a peroxidase. More than 95% of the peroxidase activity is found in the 40000 g supernatant. The total activity may be as high as 1000 times the respiratory acitivity of the callus tissue. At least two fractions, obtained by Sephadex gel filtration, can be distinguished showing this peroxidase activity, one of about 15 kDa and one 〉 50 kDa.The main properties of both fractions are:a) Hydroxamate at 0.2–0.5 mM gives half-maximal stimulation. Maximal stimulation is observed with 1–3 mM benzhydroxamate (BHAM) and 1–15 mM salicylhydroxamate (SHAM). Higher concentrations, especially of BHAM, give less or no stimulation.b) Hydroxamates are not consumed during the reaction.c) Both NADH and NADPH can serve as the electron donor for the reaction. The affinity for NAD(P)H is very low (Km near 10 mM). In the absence of hydroxamates NAD(P)H is only slowly oxidized, with an even lower affinity.d) The peroxidase can carry out two reactions: an O2-consuming and a H2O2-consuming reaction. In both reactions one NAD(P)H is consumed. In the first reaction H2O2 is formed which can be consumed in the second reaction, resulting in an overall stoichiometry of 2 NADH consumed for each O2 molecule and in the production of H2O.e) The reaction is completely blocked by cyanide, superoxide dismutase (EC 1.15.1.1) and (excess) catalase (EC 1.11.1.6), but not by antimycin A or azide. This peroxidase-mediated O2-uptake might interfere with respiratory measurements. In experiments with isolated mitochondria this interference can be prevented by the addition of catalase to the reaction mixture. The use of high concentrations of hydroxamate is not allowed because of inhibitory effects on the cytochrome pathway. In intact callus tissue hydroxamates only stimulate O2-uptake in the presence of exogenous NADH. In vivo the peroxidase does not appear to function in O2-uptake, probably because of its localization (at least partly in the cell wall) and/or its low affinity for NADH. The use of hydroxamates in the determination of cytochrome and alternative pathway activity is discussed.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 58 (1983), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Callus-forming discs from potato tubers lose 80% of their starch during one month of incubation on nutrient medium containing either 0, 3 or 6% (w/v) sucrose. The content of soluble sugar in the discs varies from 5 mg (incubated without sucrose) to 22 mg (on 3% sucrose) and 40 mg (on 6% sucrose) per g fresh weight. The activity of the cytochrome pathway (Vcyt) increases during the first week of incubation on all media. Thereafter Vcyt decreases again on 0% sucrose medium, while it remains constant on 3 and 6% sucrose media. Alternative pathway capacity (Valt), absent in freshly sliced tissue, shows a sharp increase during the first days of incubation, independent of the sucrose concentration in the medium. This capacity further increases during prolonged incubation on 3 and 6% sucrose but decreases on 0% sucrose.The in vivo activity of the alternative pathway (the participation in uninhibited respiration, ϱValt) varies with the sucrose concentration and with the culture time. In tissue incubated for 2-3 weeks on 6% sucrose as much as 45% of the electrons are transfered to oxygen via the alternative pathway. In this tissue the factor Q (the part of the alternative pathway capacity that is operative) is about 0.8, while in tissue incubated on 0 and 3% sucrose media p generally does not exceed 0.5.When chloramphenicol, an inhibitor of mitochondrial protein synthesis, is added to the medium together with 3% sucrose, the increase in Vyet does not occur, while the induction of Valt during the first week of incubation is the same as without chloramphenicol. A greater part of the alternative pathway capacity becomes operative in this tissue, leading to values of Q of almost 1 after prolonged incubation. Apparently, incubation on high sugar medium leads to extra participation in respiration of the energetically inefficient alternative oxidase pathway Excess sugar leads to wasteful respiration suggesting that the alternative oxidase functions as an ‘energy overflow’.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 62 (1984), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Potato tuber (Solanum tuberosum L. cv. Bintje) callus shows a decrease in fresh weight and an increase in dry weight upon transfer to nutrient medium supplemented with 0.3 or 0.5 M mannitol. The osmolarity of the intracellular fluid increases simultaneously. Probably mannitol is taken up from the medium till the osmolarity of the tissue is in equilibrium with that of the medium. After osmotic adaptation, on a medium with 0.5 M mannitol, growth is negligible, although the tissue retains its viability.Respiration increases upon transfer to medium with extra mannitol, especially when expressed on a fresh weight basis. On this basis cytochrome and alternative pathway capacities do not change appreciably. The respiratory increase is exclusively caused by an increased engagement of the alternative pathway. The participation of this pathway in uninhibited respiration increases from about 10 to 90% upon transfer to medium with extra mannitol. The increase in respiration is partly correlated with the decrease in fresh weight upon transfer. Per disc, the capacities of the cytochrome and alternative pathway decline. Yet, total respiration per disc significantly increases due to the increased participation of the alternative pathway. This results in an almost equal ATP-production per disc before and after transfer. We suggest, that the alternative pathway functions as a reserve capacity in potato callus, which is switched on when ATP-production coupled to the cytochrome pathway is impaired.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 7 (1986), S. 217-225 
    ISSN: 1573-5044
    Keywords: callus ; mitochondria ; oxygen ; potato ; respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Growth and oxygen uptake of potato callus is faster in an oxygen-enriched atmosphere (70% oxygen, v/v; “oxygen-callus”) than in air (20% oxygen, v/v; “air-callus”). Especially the non-mitochondrial, so-called ‘residual respiration’ is increased in “oxygen-callus”. The capacities of the mitochondrial respiratory pathways (cytochrome pathway, Vcyt and alternative pathway, Valt) are also higher in this callus. In both callus types only a small part of the alternative pathway capacity is used in uninhibited respiration. The lower oxygen uptake of “air-callus” at normal air oxygen pressures is partially due to diffusional impedance. Measurement of the respiratory parameters of “air-callus” in oxygen-saturated medium leads to higher values than measurement in air-saturated medium, although these values are still lower than those of “oxygen-callus”. ATP-production was calculated from the oxygen-uptake data and compared with the dry weight production of the callus to give values of 10.0 and 10.8 g dry weight produced.-mol ATP-1, for “air-callus” and “oxygen-callus” respectively. As no harmful side-effects are observed, cultivation of callus under elevated oxygen pressures may be useful, when rapid callus-growth is necessary.
    Type of Medium: Electronic Resource
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