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PubMed1992

Salicylic Acid induces cyanide-resistant respiration in tobacco cell-suspension cultures.

Cyanide-resistant, alternative respiration in Nicotiana tabacum L. cv Xanthi-nc was analyzed in liquid suspension cultures using O(2) uptake and calorimetric measurements. In young cultures (4-8 d after transfer), cyanide inhibited O(2) uptake by up to 40% as compared to controls. Application of 20 mum salicylic acid (SA) to young cells increased cyanide-resistant O(2) uptake within 2 h. Development of KCN resistance did not affect total O(2) uptake, but was accompanied by a 60% increase in the rate of heat evolution from cells as measured by calorimetry. This stimulation of heat evolution by SA was not significantly affected by 1 mm cyanide, but was reduced by 10 mm salicylhydroxamic acid (SHAM), an inhibitor of cyanide-resistant respiration. Treatment of SA-induced or uninduced cells with a combination of cyanide and SHAM blocked most of the O(2) consumption and heat evolution. Fifty percent of the applied SA was taken up within 10 min, with most of the intracellular SA metabolized in 2 h. 2,6-Dihydroxybenzoic and 4-hydroxybenzoic acids also induced cyanide-resistant respiration. These data indicate that in tobacco cell-suspension culture, SA induces the activity and the capacity of cyanide-resistant respiration without affecting the capacity of the cytochrome c respiration pathway.

Journal Article↗
PubMed1993

Role of Oxygen in the Limitation and Inhibition of Nitrogenase Activity and Respiration Rate in Individual Soybean Nodules.

Although infected cell O2 concentration (Oi) is known to limit respiration and nitrogenase activity in legume nodules, techniques have not been available to measure both processes simultaneously in an individual legume nodule. Consequently, details of the relationship between nitrogenase activity and Oi are not fully appreciated. For the present study, a probe was designed that allowed open circuit measurements of H2 evolution (nitrogenase activity) and CO2 evolution (respiration rate) in a single attached soybean nodule while simultaneously monitoring fractional oxygenation of leghemoglobin (and thereby Oi) with a nodule oximeter. Compared to measurements of whole nodulated roots, use of the probe led to inhibition of nitrogenase activity in the single nodules. During oximetry measurements, total nitrogenase activity (TNA; peak H2 evolution in Ar/O2) in the single nodules was 16% of that in whole nodulated roots and 48% of nodulated root activity when Oi was not being measured simultaneously. This inhibition did not affect the nodules' ability to regulate Oi, because exposure to Ar/O2 (80:20, v/v) caused nitrogenase activity and respiration rate to decline, and this decline was linearly correlated with a concurrent decrease in Oi. When the nodules were subsequently exposed to a linear increase in external pO2 from 20 to 100% O2 at 2.7% O2/min, fractional leghemoglobin oxygenation first increased gradually and then more rapidly, reaching saturation at a pO2 between 76 and 100% O2. Plots of nitrogenase activity and respiration rate against Oi showed that rates increased with Oi up to a value of 57 nM, with half-maximal rates being attained at Oi values between 10 and 14 nM O2. The maximum nitrogenase activity achieved during the increase in pO2 (potential nitrogenase activity) was 30 to 57% of that measured in intact nodulated roots, showing that O2 limitation of nitrogenase activity could account for a significant proportion of the inhibition of TNA associated with the use of the probe. However, some factor(s) in addition to O2 must have limited the activity of single nodules at both subsaturating and saturating Oi. At Oi values greater than about 57 nM, nitrogenase activity and nodule respiration were inhibited, but, because this inhibition has been shown previously to be readily reversible when the Oi was lowered, it was not attributed to direct O2 inactivation of the nitrogenase protein. These results indicate that maximum nitrogenase activity in legume nodules is supported by a narrow range of Oi values. Possible biochemical mechanisms are discussed for both O2 limitation of nitrogenase activity at low Oi and inhibition of nitrogenase activity at high Oi.

Journal Article↗
PubMed1999

Cyanide-resistant, ATP-synthesis-sustained, and uncoupling-protein-sustained respiration during postharvest ripening of tomato fruit

Tomato (Lycopersicon esculentum) mitochondria contain both alternative oxidase (AOX) and uncoupling protein as energy-dissipating systems that can decrease the efficiency of oxidative phosphorylation. We followed the cyanide (CN)-resistant, ATP-synthesis-sustained, and uncoupling-protein-sustained respiration of isolated mitochondria, as well as the immunologically detectable levels of uncoupling protein and AOX, during tomato fruit ripening from the mature green stage to the red stage. The AOX protein level and CN-resistant respiration of isolated mitochondria decreased with ripening from the green to the red stage. The ATP-synthesis-sustained respiration followed the same behavior. In contrast, the level of uncoupling protein and the total uncoupling-protein-sustained respiration of isolated mitochondria decreased from only the yellow stage on. We observed an acute inhibition of the CN-resistant respiration by linoleic acid in the micromolar range. These results suggest that the two energy-dissipating systems could have different roles during the ripening process.

Journal Article↗
PubMed1966

Relationship of lipid metabolism to the respiration and growth of pea stem sections.

Biologically active lipids increase the growth of pea stem sections within 3 hours at the same time their respiration is increased and their growth rate is more than that of the intact plant. The greater final length of the intact internode is due to a longer growth period.BOTH ACTIVE AND INACTIVE LIPIDS ARE RAPIDLY TAKEN UP AND ENTER ALL MAJOR METABOLIC FRACTIONS: among centrifugal fractions methyl oleate tends to label those that contain metabolically active membranes. It is concluded that lipids active in the bioassay are probably the effective molecules at the subcellular site of action.No direct effect of lipids on isolated mitochondria could be shown. The respiration of stem tissue was not influenced by dinitrophenol and carbonyl cyano m-chlorophenyl hydrazone although dinitrophenol inhibited growth. Lipid-induced respiration was sensitive to these agents as well as to cyanide, indicating cytochrome oxidase is probably involved.The promotion of growth and respiration by lipids is not linked to protein synthesis, since actinomycin D, puromycin and cycloheximide failed to inhibit the respiratory increase even though strongly limiting amino acid incorporation into protein. It is most likely that the effect of lipids on growth is due to their promotion of respiration.

Journal Article↗
PubMed1972

Regulation of salt respiration in carrot root slices.

In slices of carrot-phloem parenchyma washed for 7 days in water at 20 C, 50 mm KCl stimulates respiration by up to 100% of the ground respiration within 4 minutes of application. The data presented imply that ADP liberated in the cytoplasm as a consequence of KCl accumulation first stimulates a regulator reaction requiring ADP (phosphoglycerate kinase). Thereafter, the point of control alternates between this reaction and the phosphofructokinase reaction, forming a sequence of enzyme stimulations which continue after the new steady state of increased respiration is established. KCl induces a similar sequence in slices washed for 3 days, but it is completed within 3 minutes, and metabolite oscillations are not so marked. In slices washed for 2 days, KCl stimulates respiration by less than 10%, and the sequence of regulator reactions does not occur. Phosphoglycerate kinase is the only enzyme stimulated within 3 minutes of applying KCl to these slices. Contrary to previous reports, KCl frequently stimulates the respiration of freshly prepared slices by 10 to 30%.

Journal Article↗
PubMed1975

Effect of Temperature on Respiration of Mitochondria and Shoot Segments from Cold hardened and Nonhardened Wheat and Rye Seedlings.

The effect of temperature on respiration of mitochondria and tissue segments from three wheat (Triticum aestivum L.) and one rye (Secale cereale L.) cultivar grown at 2 and 24 C has been examined. Discontinuities in Arrhenius plots of respiratory activity against temperature were observed for mitochondria and tissue segments from seedlings grown at both temperatures. The rates of respiration decreased abruptly below the transition temperatures, resulting in increased energy of activation values for respiration. Transition temperatures were observed from 6 to 10 C during tissue segment respiration, and from 10 to 14 C during respiration by isolated mitochondria. Respiratory control and efficiency of phosphorylation were not affected markedly by either reaction temperature or growth temperature of the seedlings. No correlation was observed between the cold hardiness of the cultivars and the temperature at which structural transitions occurred in the mitochondria. Dry matter content of the seedlings increased markedly during growth at 2 C, but no appreciable changes in the levels of mitochondrial protein were observed. The results support the view that changes other than fatty acid unsaturation are involved in the abrupt change in mitochondrial membrane properties at low temperature.

Journal Article↗
PubMed1979

Environmental and genotypic effects on the respiration associated with symbiotic nitrogen fixation in peas.

Estimated values for the respiration associated with symbiotic nitrogen fixation in Pisum sativum L. were independent of irradiance, temperature, plant age, and CO(2) concentration, despite large variation in the total rates of C(2)H(2) reduction and root + nodule respiration. Similar values were also found in Phaseolus vulgaris L., Vicia faba L. and Glycine max (L.) Merr. Among all combinations of four Pisum cultivars with four Rhizobium leguminosarum inoculants only the plant genotype significantly affected the fixation-linked respiration, although both plant and bacterial types significantly influenced the total rate of C(2)H(2) reduction. On the basis of measured rates of H(2) evolution and C(2)H(2) reduction, or total nitrogen gain in the same system, the least respiration per unit of ammonia produced symbiotically was estimated as 4.8 to 6.9 moles CO(2) (mole NH(3))(-1) in Laxton's Progress and the greatest as 9.3 to 13.3 moles CO(2) (mole NH(3))(-1) in an Indian cultivar, as compared to a theoretical minimum respiration requirement of 4.7 moles CO(2) (mole NH(3))(-1) in peas.

Journal Article↗
PubMed1984

Inhibition and stimulation of root respiration in pisum and plantago by hydroxamate : its consequences for the assessment of alternative path activity.

The contribution of the alternative pathway in root respiration of Pisum sativum L. cv Rondo, Plantago lanceolata L., and Plantago major L. ssp major was determined by titration with salicylhydroxamate (SHAM) in the absence and presence of cyanide. SHAM completely inhibited the cyanide-resistant component of root respiration at 5 to 10 millimolar with an apparent K(i) of 600 micromolar. In contrast, SHAM enhanced pea root respiration by 30% at most, at concentrations below 15 millimolar. An unknown oxidase appeared to be responsible for this stimulation. Its maximum activity in the presence of low SHAM concentrations (1-5 millimolar) was 40% of control respiration rate in pea roots, since 25 millimolar SHAM resulted in 10% inhibition. In plantain roots, the maximum activity was found to be 15%. This hydroxamate-activated oxidase was distinct from the cytochrome path by its resistance to antimycin. The results of titrations with cyanide and antimycin indicated that high SHAM concentrations (up to 25 millimolar) block the hydroxamate-activated oxidase, but do not affect the cytochrome path and, therefore, are a reliable tool for estimating the activity of the alternative path in vivo. A considerable fraction of root respiration was mediated by the alternative path in plantain (45%) and pea (15%), in the latter because of the saturation of the cytochrome path.

Journal Article↗
PubMed1984

A Comparative Study of CN-Resistant Respiration in Different Cultures of Tobacco Callus.

The callus of Nicotiana rustica cv Gansu yellow flower and N. tabacum cv willow leaf were cultured on ordinary subculture medium (M-1) and on regeneration medium (M-2), respectively. No differentiation was observed in Gansu yellow flower tobacco callus cultures grown on both M-1 and M-2 medium. The respiration of both cultures was partially resistant to cyanide and markedly inhibited by m-chlorobenzhydroxamic acid. The relative contributions of alternative and cytochrome pathway were 31% and 47% of the total respiration, respectively, in M-1 callus cultures. The relative O(2) uptake of the two pathways was not changed significantly in M-2 callus cultures. In subcultured M-1 callus cultures of Willow leaf tobacco, the respiration mediated via alternative pathway was about 29 to 38% of the total respiration, and the cytochrome pathway still was the major respiratory pathway. In M-2 callus cultures in which differentiation occurred, the relative contribution of alternative pathway increased to 41 to 47% of the total respiration, and the cytochrome pathway decreased considerably. These results suggested that the change of respiratory electron transport pathway was probably related to the differentiation of tobacco callus cultures.

Journal Article↗
PubMed1985

Effects of CO(2) Enrichment and Carbohydrate Content on the Dark Respiration of Soybeans.

During the period of most active leaf expansion, the foliar dark respiration rate of soybeans (Glycine max cv Williams), grown for 2 weeks in 1000 microliters CO(2) per liter air, was 1.45 milligrams CO(2) evolved per hour leaf density thickness, and this was twice the rate displayed by leaves of control plants (350 microliters CO(2) per liter air). There was a higher foliar nonstructural carbohydrate level (e.g. sucrose and starch) in the CO(2) enriched compared with CO(2) normal plants. For example, leaves of enriched plants displayed levels of nonstructural carbohydrate equivalent to 174 milligrams glucose per gram dry weight compared to the 84 milligrams glucose per gram dry weight found in control plant leaves. As the leaves of CO(2) enriched plants approached full expansion, both the foliar respiration rate and carbohydrate content of the CO(2) enriched leaves decreased until they were equivalent with those same parameters in the leaves of control plants. A strong positive correlation between respiration rate and carbohydrate content was seen in high CO(2) adapted plants, but not in the control plants.Mitochondria, isolated simultaneously from the leaves of CO(2) enriched and control plants, showed no difference in NADH or malate-glutamate dependent O(2) uptake, and there were no observed differences in the specific activities of NAD(+) linked isocitrate dehydrogenase and cytochrome c oxidase. Since the mitochondrial O(2) uptake and total enzyme activities were not greater in young enriched leaves, the increase in leaf respiration rate was not caused by metabolic adaptations in the leaf mitochondria as a response to long term CO(2) enrichment. It was concluded, that the higher respiration rate in the enriched plant's foliage was attributable, in part, to a higher carbohydrate status.

Journal Article↗
PubMed1987

Cyanide-resistant respiration in photosynthetic organs of freshwater aquatic plants.

THE RATE AND SENSITIVITY TO INHIBITORS (KCN AND SALICYLHYDROXAMIC ACID[SHAM]) OF RESPIRATORY OXYGEN UPTAKE HAS BEEN INVESTIGATED IN PHOTOSYNTHETIC ORGANS OF SEVERAL FRESHWATER AQUATIC PLANT SPECIES: six angiosperms, two bryophytes, and an alga. The oxygen uptake rates on a dry weight basis of angiosperm leaves were generally higher than those of the corresponding stems. Leaves also had a higher chlorophyll content than stems. Respiration of leaves and stems of aquatic angiosperms was generally cyanide-resistant, the percentage of resistance being higher than 50% with very few exceptions. The cyanide resistance of respiration of whole shoots of two aquatic bryophytes and an alga was lower and ranged between 25 and 50%. These results suggested that the photosynthetic tissues of aquatic plants have a considerable alternative pathway capacity. The angiosperm leaves generally showed the largest alternative path capacity. In all cases, the respiration rate of the aquatic plants studied was inhibited by SHAM alone by about 13 to 31%. These results were used for calculating the actual activities of the cytochrome and alternative pathways. These activities were generally higher in the leaves of angiosperms. The basal oxygen uptake rate of Myriophyllum spicatum leaves was not stimulated by sucrose, malate or glycine in the absence of the uncoupler carbonylcyanide-m-chlorophenylhydrazone (CCCP), but was greatly increased by CCCP, either in the presence or in the absence of substrates. These results suggest that respiration was limited by the adenylate system, and not by substrate availability. The increase in the respiratory rate by CCCP was due to a large increase in the activities of both the cytochrome and alternative pathways. The respiration rate of M. spicatum leaves in the presence of substrates was little inhibited by SHAM alone, but the SHAM-resistant rate (that is, the cytochrome path) was greatly stimulated by the further addition of CCCP. Similarly, the cyanide-resistant rate of O(2) uptake was also increased by the uncoupler.

Journal Article↗
PubMed1989

Relative sensitivity of photosynthesis and respiration to freeze-thaw stress in herbaceous species : importance of realistic freeze-thaw protocols.

The relative effect of a freeze-thaw cycle on photosynthesis, respiration, and ion leakage of potato leaf tissue was examined in two potato species, Solanum acaule Bitt. and Solanum commersonii Dun. Photosynthesis was found to be much more sensitive to freezing stress than was respiration, and demonstrated more than a 60% inhibition before any impairment of respiratory function was observed. Photosynthesis showed a slight to moderate inhibition when only 5 to 10% of the total electrolytes had leaked from the tissue (reversible injury). This was in contrast to respiration which showed no impairment until temperatures at which about 50% ion leakage (irreversible injury) had occurred. The influence of freeze-thaw protocol was further examined in S. acaule and S. commersonii, in order to explore discrepancies in the literature as to the relative sensitivities of photosynthesis and respiration. As bath cooling rates increased from 1 degrees C/hour to about 3 or 6 degrees C/hour, there was a dramatic increase in the level of damage to all measured cellular functions. The initiation of ice formation in deeply supercooled tissue caused even greater damage. As the cooling rates used in stress treatments increased, the differential sensitivity between photosynthesis and respiration nearly disappeared. Examination of agriculturally relevant, climatological data from an 11 year period confirmed that air cooling rates in the freezing range do not exceed 2 degrees C/hour. It was demonstrated, in the studies presented here, that simply increasing the actual cooling rate from 1.0 to 2.9 degrees C/hour, in frozen tissue from paired leaflet halves, meant the difference between cell survival and cell death.

Journal Article↗
PubMed1989

Root Respiration and Growth in Plantago major as Affected by Vesicular-Arbuscular Mycorrhizal Infection.

Effects of vesicular-arbuscular mycorrhizal (VAM) infection and P on root respiration and dry matter allocation were studied in Plantago major L. ssp. pleiosperma (Pilger). By applying P, the relative growth rate of non-VAM controls and plants colonized by Glomus fasciculatum (Thaxt. sensu Gerdemann) Gerdemann and Trappe was increased to a similar extent (55-67%). However, leaf area ratio was increased more and net assimilation rate per unit leaf area was increased less by VAM infection than by P addition. The lower net assimilation rate could be related to a 20 to 30% higher root respiration rate per unit leaf area of VAM plants. Root respiration per unit dry matter and specific net uptake rates of N and P were increased more by VAM infection than by P addition. Neither the contribution of the alternative respiratory path nor the relative growth rate could account for the differences in root respiration rate between VAM and non-VAM plants. It was estimated that increased fungal respiration (87%) and ion uptake rate (13%) contributed to the higher respiratory activity of VAM roots of P. major.

Journal Article↗
PubMed1992

CO(2) Inhibits Respiration in Leaves of Rumex crispus L.

Curly dock (Rumex crispus L.) was grown from seed in a glasshouse at an ambient CO(2) partial pressure of about 35 pascals. Apparent respiration rate (CO(2) efflux in the dark) of expanded leaves was then measured at ambient CO(2) partial pressure of 5 to 95 pascals. Calculated intercellular CO(2) partial pressure was proportional to ambient CO(2) partial pressure in these short-term experiments. The CO(2) level strongly affected apparent respiration rate: a doubling of the partial pressure of CO(2) typically inhibited respiration by 25 to 30%, whereas a decrease in CO(2) elicited a corresponding increase in respiration. These responses were readily reversible. A flexible, sensitive regulatory interaction between CO(2) (a byproduct of respiration) and some component(s) of heterotrophic metabolism is indicated.

Journal Article↗
PubMed2002

Simulation of human respiration in fMRI with a mechanical model.

Obtaining functional magnetic resonance images of the brain is a challenging measurement process having a low characteristic signal-to-noise ratio. Images contain various forms of noise, including those induced by physiologic processes. One of the prevalent disturbances is hypothesized to result from susceptibility fluctuations caused by abdominal volume changes during respiration. To test this hypothesis and characterize the contribution of respiration noise to both magnitude and phase images, a mechanical model of a respiring human was constructed. The model was tested by comparing data from the model with that of a resting human. Power spectrum analyses show that the model induces both phase and magnitude disturbances similar to those in the human. The disturbances are directly related to the frequency of the respiration, with the noise most prevalent in the phase images. Though magnitude image noise is hard to identify in the human, the manikin demonstrates the presence of this disturbance. The construction of the manikin rules out motion as the primary source of the observed fluctuations and variation of the electrical properties of the manikin also indicates that signal fluctuations are not primarily due to eddy currents. Therefore, the changes are most probably induced by bulk susceptibility changes correlating with respiration.

Artifacts↗
PubMed1984

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.

Binding Sites↗
PubMed1985

Decrease in respiration activity related to prodigiosin synthesis in Serratia marcescens.

Variation in the cell respiration rate of pigmented and nonpigmented strains of Serratia marcescens was exhibited. The respiration rate of a pigmented strain decreased earlier than that of nonpigmented strains in the late exponential or early stationary phase. However when prodigiosin synthesis was not induced by exchange of carbon sources in the medium, the decrease in the respiration rate of the pigmented strain was the same as that of nonpigmented strains. Measurement of the oxygen consumption rate in the sonicated cell membrane by adding NADH solution showed that the rate in the pigmented strain was lower than that in nonpigmented strains. Furthermore, the cell membrane of prodigiosin-induced organisms was more sensitive to respiration inhibitors than that of pigment-noninduced organisms of the pigmented strain. These results showed that the respiration activity was decreased by prodigiosin synthesis in S. marcescens.

Cell Fractionation↗
PubMed1989

The long-term effect of a positive pressure respirator on the specific antibody response in pigeon breeders.

A group of pigeon fanciers with Pigeon Breeder's Lung wore a positive pressure respirator with a full mask and hood when working with pigeons. Serial measurements of specific IgG antibody to pigeon globulin (ELISA) were made in 22 subjects. Thirteen wore a respirator (monitored over 14 months) and nine did not (monitored over 3 months). Following introduction of the respirator, the serum antibody to pigeon globulin significantly decreased when compared with the non-respirator group. This fall in antibody persisted over 14 months reflecting the reduction of antigen inhalation. Although circulating antibody decreased in all those wearing respirators, the remaining levels were still within the range that can be associated with clinical symptoms possibly because of inward leakage of antigen bypassing the filter.

Adult↗