Effect of High Temperature on the Development of Cyanide-Sensitive Respiration in the Germination Process of Spinach Seeds

Germination of spinach (Spinacia oleracea L. var. grabra cv.Nobel) seeds was inhibited at a high temperature (35?C). Effectsof KCN on the respiration of seeds incubated at 20 and 35?Cwere compared in order to investigate the mechanism of inhibitionof seed germination by high temperature. Respiration of germinatingseeds incubated at 20?C was inhibited about 50% by 5 mM. KCNsolution, whereas it hardly inhibited the weak respiration ofthe seeds at 35?C. Germination of seeds was delayed by exogenousKCN. When the KCN solution was renewed daily, germination wascompletely inhibited. Pericarp removal promoted germinationat 35?C, but atypical germination (cotyledons emerging earlierthan a radicle) took up more than half of the total germination.The inhibitory action of KCN on the respiration of seeds wasnot altered by pericarp removal. A KCN addition, even at 20?C,elicited atypical germination in the pericarp-less seeds. Theseresults show that cyanide-sensitive respiration is needed toinduce typical spinach seed germination (root emergence), butis rendered inoperative by high temperatures thus bringing aboutpoor germination and atypical germination. (Received December 1, 1984; Accepted February 8, 1985)     

Action of Respiratory Inhibitors on Seed Germination and Oxygen Uptake in Avena fatua L.

The effects of sodium azide, potassium cyanide (cytochrome oxidaseinhibitors), and salicylhydroxamic acid (SHAM; an alternativerespiration inhibitor) on germination and respiration of Avenafatua L. seeds were studied. Azide and cyanide released seeddormancy at similar concentrations and treatment durations.Cyanide, however, stimulated germination of seeds with littleafter-ripening, whereas azide had no effect under similar conditionsunless the seeds were after-ripened for several months; theduration of after-ripening required for seeds to respond toazide varied with seed batch. There was also a greater lag priorto germination in the case of azide, compared to cyanide treatedseeds. SHAM inhibited the stimulation of germination and respirationby azide, but not by cyanide. Furthermore, respiration induced by azide or cyanide could notbe inhibited by the subsequent application of SHAM. These findingssuggest that the respiration stimulated by azide and cyanideis not alternative (SHAM-sensitive) and, therefore, this respiratorypathway cannot be involved in the stimulation of germinationby cytochrome oxidase inhibitors. While embryos excised fromcontrol, azide or cyanide pretreated seeds had the capacityto perform alternative respiration, the actual contributionof this pathway was negligible. A large proportion of respirationof embryos excised from azide or cyanide pretreated seeds wasresidual, i.e. insensitive to both SHAM and cyanide. Alternative respiration, azide, cyanide, dormancy, salicylhydroxamic acid, wild oats     

Alteration of the respiratory function in chill-sensitive callus due to low temperature stress: Involvement of the alternate pathway

The diversion of electron flow to the alternate pathway in mitochondriaof chillsensitive callus of Cornus stolonifera was studied withreference to the immediate response of the cells to chilling.Temperatures below 15?C diverted the bulk of respiration tothe alternate path, proportionally as the temperature decreasedto 0?C. The alternate path, however, existed in cells in a suppressedform at temperatures above 15?C. In chill-resistant callus ofSambucus Sieboldiana, no alteration was noted in electron apportionmentbetween the cytochrome path and the alternate path. Thus, thereseems to be a marked difference in the mode of respiration betweensensitive and resistant plant cells at low temperatures. Therespiratory control ratio also markedly declined at temperaturesbelow 15?C in chill-sensitive callus, but a higher respiratorycontrol ratio was observed below 10?C in chill-resistant callus.From these results, an alteration in the regulatory system forelectron apportionment between the two paths is probably themost immediate response of chill-sensitive cells to low temperatures. ¹Contribution No. 2154 from the Institute of Low TemperatureScience. (Received June 6, 1979; )     

Pea Fruit Extension Rate: II. EFFECT OF TEMPERATURE ON THE RELATIONSHIP WITH RESPIRATION

Rates of respiration and extension of pea fruits maintainedunder different regimes of fruit (10 ?C to 25 ?C) and plant(15 ?C to 30 ?C) temperature were measured during prolongeddarkness. Higher fruit temperatures resulted in faster maximumrates while higher plant temperatures were associated with slowermaximum rates of fruit extension and respiration. Increasingthe temperature of either part caused faster decay of ratesin the dark; this effect being more apparent for respirationthan for growth in length and for plant temperatures than forfruit temperatures. The relationship between maximum rates ofgrowth and respiration for different temperatures was linear.There was an asymptotic curvilinear relationship between specificrespiration and relative extension rates obtained during prolongeddarkness. Analysis of these data using an exponential equationof the form showed that treatment regimes affected primarily the asymptote(A) of this equation. Estimates of the shape parameters (b andk) of the curve were very variable and no obvious treatmenteffects could be discerned. The form of these data is comparedwith those of other models for respiration. Key words: Peas, Fruits, Growth, Extension, Respiration, Temperature, LVDT     

Root Respiration for Agave deserti: Influence of Temperature, Water Status and Root Age on Daily Patterns

Root respiration, measured as CO₂ efflux, was studied for asucculent perennial from the Sonoran Desert, Agave deserti,with a new technique using individual, attached roots. The dailypatterns of root respiration closely followed the daily patternsof root temperature for both established roots and rain roots,with higher rates during the day when root temperature averaged27?C and lower rates at night when root temperature averaged17?C. When root temperature was raised from 5?C to 40?C, rootrespiration increased about 7-fold; from 45 ?C to 55 ?C, rootrespiration decreased about 2-fold, except for old establishedroots. Root respiration per unit dry weight for both root typesdecreased with age, the initial decrease being greater for rainroots than for established roots. Root respiration rates forrain roots were reduced to zero at a soil water potential (_soil)of –0.9 MPa and did not recover upon rewatering. Upondrying, root respiration rates for established roots were maintainedat about 12% of maximum, even when _soil fell to –1.6 MPa,and fully recovered 1.5 d after rewatering the soil. Such responsesof rain and established roots must be taken into account whenassessing the carbon costs for the root system. Key words: Agave deserti, CO₂ exchange, root respiration, temperature, soil water potential     

Metabolism of urea in Chlorella ellipsoidea

The effect of cyanide on ammonia and urea metabolism was studiedwith intact cells of Chlorella ellipsoidea Gerneck, a greenalga which apparently lacks urease. Ammonia uptake was inhibited more readily by cyanide than wasurea uptake. Urea uptake was stimulated by lower concentrationsof cyanide. The addition of cyanide caused the formation ofammonia from some cellular nitrogenous compounds. In the presenceof exogenously added urea, the molar ratio of ammonia accumulatedin the medium to urea taken up exceeded 2.0 as the cyanide concentrationincreased. However, the molar ratio of ammonia actually producedfrom urea nitrogen to urea taken up was less than 1.35 at anyconcentration of cyanide tested. In the presence of higher concentrationsof cyanide, the rate of incorporation of ¹⁵N into amino acidsfrom ¹⁵N-urea was higher than that from ¹⁵N-ammonium sulfate. The results suggest that Chlorella ellipsoidea possesses a pathwaythrough which urea nitrogen is assimilated directly withouta preliminary breakdown to ammonia. (Received October 18, 1976; )     

The Effect of Cyanide on the Respiration and the Oxidative Assimilation of Glucose by Chlorella vulgaris

It is shown that low concentrations of cyanide stimulate theendogenous respiration of Chlorella vulgaris. When glucose isadded the respiration rate is much increased but is now reversiblyinhibited by cyanide. Some 30–60 per cent. of the totalrespiration remains uninhibited. One-eighth to one-ninth ofthe glucose added is completely oxidized. Most of the remainderis assimilated to di- or polysaccharide. Low concentrationsof cyanide which inhibit the rate of glucose oxidation alsoinhibit the assimilation of glucose. Two possible interpretationsof this fact are discussed. It is suggested that the assimilationof glucose is coupled with the oxidation of glucose by a cyanide-sensitiverespiratory system. The mathematical consequences of this theoryare considered and shown to agree with the experimental results.The effect of cyanide on the respiratory quotient is also discussed.     

Light Actions in the Germination of Cocklebur Seeds: IV. DISAPPEARANCE OF RED LIGHT-REQUIREMENT FOR THE GERMINATION OF UPPER SEEDS SUBJECT TO ANOXIA, CHILLING, CYANIDE OR AZIDEPRETREATMENT

Esashi, Y., Fuwa, Nn Kojima, K. and Hase, S. 1986. Light actionsin the germination of cocklebur seeds. IV. Disappearance ofred light-requirement for the germination of upper seeds subjectto anoxia, chilling, cyanide or azide pretreatmenL—J.exp. Bot. 37: 1652–1662. The effects on the germination of positively photoblastic uppercocklebur (X anthium pennsylvanicum Wallr.) seeds by pretreatingwith anoxia, chilling, cyanide or azide, which stimulates theirdark germination, were examined in relation to light actions.Prior to experiments, seeds were pre-soaked at 23 °C inthe dark for 1 or 2 weeks to remove the pre-existing Pfr. Whenthe prctreatment conditions were suboptimal for germinationinduction, the stimulating effects of the pretreatments on germinationduring a subsequent dark period at 23 °C were manifest onlywhen seeds were irradiated with red light before or after thepretreatment Red light promotion was reversed by blue or far-redlight treatment. However, both prc-chilling for 6 d at 8 °Cand prctreatment with 1· 5 mol m ^{– 3} NaN³ for 2d could induce full germination without red light exposure.On the other hand, both pre-exposure to anoxia for 8 d and pretreatmentwith 30 mol m^–3 KCN could induce the dark germinationonly when germination occurred at 33 °C which is known toaugment the ratio of an alternative respiration flux to a cytochromeone. Moreover, the dark germination in response to these inductionswere strongly inhibited by the inhibitors of alternative respiration,propyl gallate and benzohydroxamic acid, applied during a subsequentdark period. It was thus suggested that Pfr has some relationto the operation of two respiration systems of cocklebur seeds,but it is not indispensable to germination of this positivelyphotoblastic seed. Key words: Anoxia, azide, blue light, chilling cyanide, dark germination, far-red light, red light, seed germination, X anthium pennsylvanicum     

The Effects of Low Growth and Measurement Temperature on the Respiratory Properties of Five Temperate Species

Five temperate ruderal species, Epilobium glandulosum, Matricariamatricariodes, Melilotus alba, Sonchus asper and Taraxacum officinale,were grown under controlled conditions at both 10 °C and20 °C. Respiration was measured at 10 °C and 20 °C.Total dark respiration rates of plants grown at 10 °C wereconsistently higher than the rates of plants grown at 20 °C.This was due to greater respiratory activity along both thecytochrome (KCN-sensitive) and alternative (SHAM-sensitive)pathways in the cold grown plants. The capacity to use the alternativepathway was also greater in the 10 °C grown plants. Theincrease in the relative contribution of each pathway was speciesspecific. At a given growth temperature the activity of thealternative pathway varied little between measurements at 10°C and 20 °C. In contrast, the activity of the cytochromepathway was greater when measured at 20 °C than when itwas measured at 10 °C. The capacity of the alternative pathwaywas lower when measured at 10 °C. These results suggestthat the cytochrome pathway is more sensitive to temperaturestress than is the alternative pathway. Alternative pathway, cyanide resistance, cytochrome pathway, respiration, temperature     

Characteristics of light-induced 515-nm absorbance change in spinach chloroplasts at lower temperatures I. Participation of structural changes of thylakoid membranes in light-induced 515-nm absorbance change

Light-induced absorbance change at 515 nm in spinach chloroplastswas studied in the temperature range from –2?C to 27?C.Lowering of temperature had no marked effect on the extentsof initial "light-on" spike and the steady-state change overthe temperature range examined, whereas the rate of recoveryof the 515-nm change was significantly reduced at lower temperatures.Above 15?C, recovery of the 515-nm change after continuous illuminationshowed a first-order kinetics. In contrast, the recovery wascomposed of a fast and a slow phases at lower temperatures. The fast phase of the recovery of the 515-nm change was acceleratedby carbonyl cyanide m-chlorophenylhydrazone, valinomycin plusK⁺ or sodium tetraphenylboron, while the slow phase was completelyeliminated in glutaraldehyde-fixed chloroplasts. Light-inducedchange in absorbance at 546 nm, an indicator of structural changesof membrane, showed almost the same dependency on temperatureas the slow phase of the recovery of the 515-nm change. Theseresults suggest that not only electric field formation acrossthe thylakoid membrane but also structural or conformationalchanges in the membrane participate in the 515-nm absorbancechange observed under steady illumination. (Received July 5, 1976; )     

Influences of Water Status, Temperature, and Root Age on Daily Patterns of Root Respiration for Two Cactus Species

Daily patterns of root respiration measured as CO₂, efflux werestudied at various soil water potentials, temperatures, androot ages for individual, attached roots of the barrel cactusFerocactus acanthodes and the platyopuntia Opuntia ficus-indica.The daily patterns of root respiration for both establishedroots and rain roots followed the daily patterns of root temperature.Root respiration increased when root temperature was raisedfrom 5 °C to 50 °C for F. acanthodes and from 5 °Cto 55 °C for O. ficus-indica; at 60 °C root respirationdecreased 50° from the maximum for F. acanthodes and decreased25° for O. ficus-indica. Root respiration per unit d. wtdecreased with root age for both species, especially for rainroots. Root respiration rates for rain roots were reduced tozero at a soil water potential (     

Metabolic Inhibitors Block ABA-Induced Stomatal Closure

Closure of stomata of Commelina communis L. leaf epidermis causedby abscisic acid (ABA) was inhibited by sodium azide, potassiumcyanide and hypoxic conditions. Azide was more effective thancyanide at low concentrations, but the cyanide effect couldbe enhanced by addition of salicylhydroxamic acid, providingindirect evidence for cyanide-resistant respiration in epidermaltissue. Azide also inhibited ABA-induced closure of ‘isolated’ stomata and shrinkage of guard cell protoplasts.The results indicate that metabolic energy is required for ABAaction involving solute loss from the guard cells. Possiblemechanisms of action are discussed.     

Effect of Temperature on Nitrogenase Activity in White Clover

Single, clonal plants of white clover were grown without inorganicnitrogen in four contrasting day/night temperature regimes,with a 12 h photoperiod, in controlled environments. Root andnodule respiration and acetylene reduction activity were measuredin a flow-through system during both day and night for plantsacclimated to day/night regimes of 23/18, 15/10 and 10/5 ?C.Similar measurements were made on plants acclimated to 20/15?C and stepwise at temperatures from 4 to 33 ?C. Peak rate of ethylene production, nitrogenase-linked respirationand basal root + nodule respiration increased approximatelylinearly from 5 to 23 ?C both in temperature-acclimated plantsand in plants exposed to varying measurement temperatures. Themeasured attributes did not vary significantly between day andnight. Temperatures above 23–25 ?C did not further enhancethe rate of ethylene production, which remained essentiallythe same up to the maximum measured temperature of 33 ?C. The measurements of nitrogenase-linked respiration between 5and 23 ?C, during both day and night, demonstrated a constant‘energetic cost’ of acetylene reduction of 2.9 µmolCO₂ µmol C₂H₄^–1,. Over the same temperature range,the approximate activation energy of acetylene reduction was60 kJ mol^–1. The integrated day plus night nitrogenase-linkedrespiration accounted for 13.4–16% of the plant‘snet shoot photosynthesis in a single diurnal period: there wasno significant effect of temperature between 5 and 23 ?C. Key words: Trifolium repens, white clover, temperature, N₂ fixation, respiration     

The alternate respiratory pathway of Candida albicans

Candida albicans contains a cryptic cyanide and antimycin A insensitive respiratory system. This alternate oxidase was found (i) at all growth rates from =0.05 to 0.26 in a chemostat culture and (ii) in both mycelial and yeast forms of the organism. Neither chloramphenicol nor cycloheximide prevented the expression of the alternate oxidase. Salicyl-hydroxamic acid was a potent inhibitor of the cyanide insensitive respiration. The respiration of mitochondria grown in the presence of antimycin A was not inhibited by cyanide or antimycin A but was inhibited by salicylhydroxamic acid.Abbreviations KCN potassium cyanide - SHAM salicyl hydroxamic acid     

Characterization of membrane-bound Mg++-activated ATPase isolated from the lower epidermis of tobacco leaves

Membrane-bound Mg⁺⁺-activated ATPase was separated from thelower epidermis of tobacco leaves (Nicotiand tabacum L. SamsunNN) on stepwise sucrose density gradient centrifugation. Membrane-bound epidermal ATPase was localized in the interfaceof densities in sucrose of 1.12 to 1.16 in the sedimentary fractionbetween 1,500?g to 10,000?g from the homogenate of the lowerepidermis. The epidermal ATPase activity was activated by divalentcations (Mg⁺⁺>Mn⁺⁺Co⁺⁺>Fe⁺⁺>Zn⁺⁺>Ca⁺⁺) and furtherstimulated by KCl by ca. 20%. The pH optimum for Mg⁺⁺-activationof the epidermal ATPase was ca. 6.0. The enzyme hydrolyzed ATPmore rapidly than other nucleoside triphosphates. The optimumtemperature for activation of the epidermal ATPase activitywas ca. 40?C. 50% of the epidermal ATPase activity was lostin 18 min at 55?C and in 2.5 days at 2.5?C. The apparent Kmvalue of the epidermal ATPase was 4.7?10^–4 M and Vmaxwas 65.4 nmoles Pi/mg protein/min. The epidermal ATPase wasstrongly inhibited by N, N'-dicyclohexylcarbodiimide (DCCD)in vitro whereas oligomycin, carbonyl cyanide m-chlorophenylhydrazone(CCGP), indoleacetic acid (IAA) and abscisic acid (ABA) wereinsensitive to the epidermal ATPase activity. (Received May 23, 1978; )     

Long-term Respiratory Cost of Maintenance and Growth of Field-grown Young Hinoki Cypress (Chamaecyparis obtusa)

Respiration rate of the entire above-ground parts of field-grown8-year-old hinoki cypress [Chamaecyparis obtusa(Sieb. et Zucc.)Endl.] was measured at monthly intervals over a 5-year period,to evaluate the trend in proportion of maintenance and growthcomponents of respiration with stand development. Representativesample trees were selected for respiration measurements. Theannual respiration rates of individual sample trees were combinedand partitioned into maintenance and growth components by regressingspecific respiration rate on mean relative growth rate. Maintenanceand growth respiration coefficients generated in this way were5.2 mol CO₂kg^-1year^-1and 39 mol  CO₂kg^-1, which are equivalentto 14.3 mg C kg^-1C h^-1(at mean annual air temperature of 15.1°C) and 0.94 kg C kg^-1C. Considering the maintenance andgrowth respiration coefficients, and phytomass and phytomassincrement of individual trees in the stand, the maintenanceand growth respiration rates of the stand were estimated. Theproportion of the maintenance respiration increased, whereasthat of the growth respiration decreased with stand development,due to decreasing relative growth rate.Copyright 1997 Annalsof Botany Company Chamaecyparis obtusa; growth respiration coefficient; hinoki; maintenance respiration coefficient; stand respiration     

δ-N-[methyl-14C]arginine: A simplified preparation

A method is described for quantitative assay of cyanide which is based on measuring the rate of rise in absorbancy at 325 nm when cyanide forms an addition compound with NAD⁺ at pH 10. It was found that the rate of rise in absorbancy is proportional with the concentration of cyanide within the range of 0.05–4.00 mm. By applying this method it could be shown that in biological systems in which the interface between the aqueous phase and the atmosphere is large the effective concentration of cyanide, added to inhibit the respiration, decreases rather rapidly during incubation.     

Variation in Photorespiration in Lolium

The rate of photorespiration in several grass species was shownto be highly variable and dependent on the species, genotype,and conditions under which the plants were grown. Photorespiration,measured as oxygen uptake, was negligible in Cenchrus ciliarisand Paspalum dilatatum but significant in Lolium spp. and Festucaarundinacea. There were significant differences in the rateof photorespiration among plants within a Lolium populationof diverse origin and these differences were independent ofthe conditions under which the plants were grown. Among thetemperate grasses there was a significant correlation betweenphotorespiration and the CO₂-compensation concentration andboth parameters were very low in P. dilatatum. Plants grownin day/night temperatures of 15/10 °C compared with 25/20°C had faster rates of dark respiration but slower ratesof light respiration when measured at the same temperature.Photorespiration was faster than dark respiration although differencesin respiration among plants in the light were not shown in thedark.     

An Analysis of the Relationship between Respiration and the Transmembrane Potential in Corn Roots

The effects of cyanide, anoxia, and temperatures varying from 2 to 42 C on the cell membrane electropotential difference (PD) of washed and freshly excised corn roots have been determined. Respiration rates of freshly excised root segments in response to cyanide and to varying temperatures were also measured. The cell membrane PD of roots which had been washed for 12 to 15 hours was almost insensitive to cyanide and anoxia but sensitive to low temperature. In contrast, the cell membrane PD of freshly excised roots was reversibly depolarized by all three treatments, cyanide depolarized from −117 to −86 millivolts and the sequential imposition of anoxia further lowered the PD to −69 millivolts. Anoxia applied first depolarized maximally and the PD was not further lowered by sequential cyanide treatment. Arrhenius plot analysis of the temperature response of respiration showed an apparent transition at 13 C with an activation energy of 20.0 kilocalories per mole below and 8.8 kilocalories per mole above the transition temperatures. The energy of activation for repolarization of PD is much higher; 53.4 kilocalories per mole below 7 to 8 C and 25.4 kilocalories per mole above this apparent transition. The energy requirement for polarization of the cell membrane PD was calculated based on the temperature responses of the cell membrane PD and respiration. It was estimated that 3.5% of the energy output from respiration at 22 C is required for cell polarization. It is unlikely that ion transport is limited by energy availability below the 8 C transition in this chill sensitive species.     

Some Specific Features of the Respiration of Hydrocarbon-utilizable Candida Yeasts

The respiration of both glucose-grown and hydrocarbon-grown cells of Candida tropicalis pK 233 harvested in the stationary phases was not inhibited by cyanide when glucose was used as oxidation substrate, but the former was rather stimulated in the presence of cyanide. When n-alkanes were used as oxidation substrate, cyanide lowered the respiratory activities of both cells to about 50%. With respect to the susceptibility to cyanide, the younger cells growing on n-alkanes were less sensitive in hydrocarbon oxidizing ability than the older cells, whereas the older cells growing on glucose or n-alkanes were more resistant in glucose oxidizing ability than the younger cells. Acetate was oxidized by both glucose-grown and hydrocarbon-grown cells of the yeast. Laurate was oxidized by hydrocarbon-grown cells, but not by glucose-grown cells. The respiration on laurate was inhibited completely by 3.3 mM of cyanide. In general, hydrocarbon-grown cells of Candida tropicalis pK 233 were more sensitive to various respiratory inhibitors than glucose-grown cells, although the oxidation substrates had a significant effect.

The respiration of both glucose-grown and hydrocarbon-grown cells of C. albicans, C. guilliermondii and C. lipolytica harvested in the stationary phases was also resistant to cyanide when glucose was used as oxidation substrate. But the respiration on n-alkanes of these cells was inhibited significantly by 3.3 mM of cyanide except for C. albicans.