Some Observations on a Circadian Rhythm in Carbon Dioxide Compensation in Bryophyllum fedtschenkoi

Detached shoots of Bryophyllum fedtschenkoi maintained in continuouslight and at a constant temperature exhibit a circadian rhythmin CO₂ compensation. The rhythm has a period of 21.6±0.1h at 23 °C and its phase can be set and shifted by suitabletreatments. These observations suggest that the ryhthm is trulyendogenous in nature. The phase is set by a light-on stimulus, or a drop in lightintensity. Phase shift can be induced by short periods (3 h)of reduced light intensity as well as similar periods of darknessgiven during the day phase of the rhythm. A change from whitelight to the same number of incident quanta of monochromaticlight of 450 nm (blue), 550 nm (green), or 652 nm (red) alwaysshifted the phase, but the degree of shift was less in red lightthan in blue or green. The plant reacted to achange to blueor green light as though it had been placed in the dark. Althbughred light was apparently ‘recognized’, the changefrom white to red was still sufficient to alter the phase ofthe rhythm. The possible role of phosphoenolpyruvate carboxylase activityin controlling the rhythm is discussed.     

An Endogenous Rhythm in the Rate of Carbon Dioxide Output of Bryophyllum: I. SOME PRELIMINARY EXPERIMENTS

A techinique is described for recording automatically, withthe aid of an infrared gas analyzer, the rate CO₂ output orabsorption by plant material under controlled conditions. An examination of the rate of CO₂ output by excised leaves of16 species of succulent plants in darkness and in a CO_-2-freeatmosphere revealed clearly defined rhythms in only Bryophyllumfedtschenkoi, B. daigremontianum and B. calycinum (pinnatum). Further investigation of the rhythm in leaves of B. fedtschenkoirevealed that: (1) daylength has no effect upon the period ofthe rhythm in subsequent darkness, the phase being set at thetime the lights are extinguished; (2) normal air suppressesthe rhythm; (3) removel of the epidermis and cutting the mesophyllinto pieces 1 cm² does not effect either the phase or periodof the rhythm; (4) continuous illumination at an intensity of3,000 lux inhibits the rhythm which restarts when the lightsare extinguished. The phase of the rhythm can be set at anytime of day according to the time at which the lights are extinguished.The time which elapses between the onset of darkness and thefirst peak decreases as the length of the light treatment isincreased. The endogenuous nature of the rhythm is fully established. Theresults are compared with of other researches.     

A Circadian Rhythm in the Level of the Carbon Dioxide Compensation Point in Bryophyllum and Coffea

Measurements of CO² compensation were made over long periodsof time for Coffea arabica and Bryophyllum daigremontianum.For each species there was a marked rhythm in the level of CO²compensation, even though all major factors of the environmentwere held as constant as possible. The rhythm in Coffea wassusceptible to phase-shift and had a period slightly greaterthan 24 h. The rhythm in Bryophyllum had a period of c. 23 h.These observations show that the rhythm is probably endogenousin nature.     

Carbon Dioxide Efflux from Leaves in Light and Darkness

Efflux of carbon dioxide in light and darkness was measured at low ambient CO₂ concentrations in leaves of Rumex acetosa. Light carbon dioxide production (photo-respiration) was found to depend on irradiance and to differ from dark production as to the response to temperature and ambient concentrations of O₂ and CO₂. These observations support previously made suggestions that photorespiration follows a different metabolic pathway to dark respiration.     

An Endogenous Circadian Rhythm in the Rate of Carbon Dioxide Output of Bryophyllum: VI. ACTION SPECTRUM FOR THE INDUCTION OF PHASE SHIFTS BY VISIBLE RADIATION

A comparison has been made of the effectiveness of a 4-h exposureto equal quantum-flux densities of radiation in different zonesof the visible spectrum in shifting the phase of the endogenouscircadian rhythm of carbon dioxide metabolism in leaves of Bryoyihyllumfedtschenkoi. At an incident quantum-flux density of 8.9 ? 10^–13einsteins cm^–2 8^–1, only radiation between 600 and700 nm induced a phase shift, maximum activity being found at(560 nm. At a higher incident quantum-flux density of 1.9 ?10^–11 einsteins cm^–2 8^{– 1}, the peak of activitywas broader and extended from 560 to 700 nm. At both flux densitiesa sharp cut-off occurred at 700 nm. The action spectra are somewhat similar to the absorption spectrumof phytochrome except that they show no minor peak in the bluezone. No evidence has yet been obtained that the inductive effectof red light can be reversed by exposure to far-red radiationas in the case of a typical phytochrome-mediated response. Ultra-violet radiation at 237 nm had no effect upon the phaseof the rhythm.     

The Damping and Reinitiation of the Circadian Rhythm of CO2 Output in Bryophyllum Leaves in Relation to their Malate Content

The circadian rhythm of CO₂ output in leaves of Bryophyllumfedtschenkoi damps out after 3–4 d in continuous darknessand a CO₂-free air stream at 15°C. The rhythm is reinitiatedafter a single exposure to white light of 2, 4, 6 or 8 h duration,damps out again after a further 3–4 d and can be reinitiatedfor a second time by a further exposure to light. During the exposure to light there is a burst of CO₂ outputconsistent with the decarboxylation of malate, and the rhythmbegins afterwards with an initial high rate of CO₂ fixation.Malate gradually accumulates in the leaves in continuous darknessto attain a maximum value (35 mol m^–3) at the time whenthe circadian rhythm disappears, and decreases to a low value(19 mol m^–3) after a 4 h exposure to light which reinitiatesrhythmicity. These results support the hypothesis that damping of the rhythmof CO₂ output in continuous darkness is due to the accumulationof malate in the leaf cells, eventually reaching such a levelthat its removal from the cytoplasm into the vacuole cannottake place, with the result that PEPc activity, upon which therhythm of CO₂ output depends, remains allosterically inhibited. Key words: CAM, circadian rhythm, Bryophyllum, CO₂-fixation, malate metabolism     

A rapid circadian rhythm of carbon-dioxide metabolism in Bryophyllum fedtschenkoi

Leaves of Bryophyllum fedtschenkoi Hamet et Perrier maintained in a stream of normal air and at 15° C exhibit a circadian rhythm of CO₂ uptake in continuous light but not in continuous darkness. The rhythm is unusual in that it persists for at least 10 d, and has a short period of approximately 18 h. The mechanism by which this rhythm is generated is discussed.Abbreviation PEPCase phosphoenolpyruvate carboxylase     

The role of temperature in the regulation of the circadian rhythm of CO2 fixation in Bryophyllum fedtschenkoi

Detached leaves of Bryophyllum fedtschenkoi Hamet et Perrier kept in normal air show a single period of net CO₂ fixation on transfer to constant darkness at temperatures in the range 0–25 °C. The duration of this initial fixation period is largely independent of temperature in the range 5–20 °C, but lengthens very markedly at temperatures below 4 °C, and is reduced at temperatures above 25 °C. The onset of net fixation of CO₂ on transfer of leaves to constant darkness is immediate at low temperatures, but is delayed as the temperature is increased. The ambient temperature also determines whether or not a circadian rhythm of CO₂ exchange occurs. The rhythm begins to appear at about 20 °C, is most evident at 30 °C and becomes less distinct at 35 °C. The occurrence of a distinct circadian rhythm in CO₂ output at 30° C in the absence of a detectable rhythm in PEPCase kinase activity shows that the kinase rhythm is not a mandatory requirement for the rhythm of PEPCase activity. However, when it occurs, the kinase rhythm undoubtedly amplifies the PEPCase rhythm.Abbreviation PEPCase phosphoenolpyruvate carboxylase We thank the Agricultural and Food Research Council for financial support for this work.     

Mechanism of Excitation of Aplysia Neurons by Carbon Dioxide

The abdominal ganglion of Aplysia californica was perfused with artificial seawater equilibrated at different P_COCO2's and pH's for 5 min or less. 5% CO₂ dropped perfusate pH from 8.0 to 6.5 and produced depolarization and increased discharge rate in visceromotor neurons. Half the giant cells studied had a similar response, whereas the other half were hyperpolarized. Pacemaker neurons showed little, if any, response to such changes in pH or CO₂. Membrane conductance of responsive cells was always increased. The effect of CO₂ occurred even when synaptic transmission was blocked by low calcium and high magnesium, and therefore must have been a direct result of CO₂ or the concomitant fall in pH. When extracellular pH was lowered to 6.5 using HCl or H₂SO₄ and no CO₂, the same effects were observed. Also, local application of HCl or H₂SO₄ to the external surface of the cell soma elicited depolarization and spike discharge. When extracellular pH was held constant by continual titration, 5–50% CO₂ had no effect. Intracellular pH was probably decreased at least one pH unit under these circumstances. Thus CO₂ per se, decreased intracellular pH, and increased bicarbonate ion were without effect. It is concluded that CO₂ acts solely through a decrease in extracellular pH.     

Rapid Phase Resetting of a Mammalian Orcadian Rhythm by Brief Light Pulses

The phase-shift (Δψ) responses of the circadian rhythm in the field mouse Mus booduga to brief light pulses (LPs) of 15 minutes duration and 1000 lux intensity were measured in 90 experiments. In each experiment, a resetting light pulse LP₁ was administered at CT14 (CT, circadian time), and a scanning light pulse LP₂ was then variously administered in separate experiments at CT16, CT20, and CT22 in the same and in the next circadian cycle. The Δψ obtained in all these two-pulse experiments did not differ significantly from theoretical values computed on the assumption that LP₁ reset the phase response curve (PRC) rapidly. In each case, the steady-state Δψ observed after LP₁ and LP₂ differed significantly from the Δψ obtained at the same CT in determination of the single-pulse PRC (control) and also differed significantly from the values on the assumption of no Δψ in the PRC following LP₁. These results indicate that the circadian pacemaker of M. booduga, as measured by its PRC, is substantially reset within 2h after a light pulse at CT14. (Chronobiology International 14(6), 537–548, 1997)     

Precise Measurements of Carbon Dioxide Exchange by Illuminated Leaves near the Compensation Point

The results of precise measurements of rates of carbon dioxideassimilation at low external concentrations of carbon dioxideand of rates of carbon dioxide output into virtually carbondioxide-freeair show that linear extrapolations of the carbon dioxide intakecurve plotted against the external concentrations of carbondioxide give an exaggerated estimate of the rate of carbon dioxideoutput from illuminated leaves into carbon dioxide-free air. The shape of the exchange curve suggests that the rate of endogenousproduction of carbon dioxide changes at external concentrationsin the region of the carbon dioxide compensation point ().     

Period and phase control by temperature in the circadian rhythm of carbon dioxide fixation in illuminated leaves of Bryophyllum fedtschenkoi

The rhythm of CO₂ assimilation exhibited by leaves of Bryophyllum fedtschenkoi maintained in light and normal air occurs only at constant ambient temperatures between 10°C and 30°C. Over this range the period increases linearly with increasing temperature from the extremely low value of 15.7 h to 23.3 h, but shows a considerable degree of temperature compensation. Outside the range 10°C–30°C the rhythm is inhibited but re-starts on changing the temperature to 15°C. Prolonged exposure of leaves to high (40°C) and low (2°C) temperature inhibits the rhythm by driving the basic oscillator to fixed phase points in the cycle which differ by 180°, and which have been characterised in terms of the malate status of the leaf cells. At both temperatures loss of the circadian rhythm of CO₂ assimilation is due to the inhibition of phosphoenolpyruvate carboxylase (PEPCase) activity, but the inhibition is apparently achieved in different ways at 40°C and 2°C. High temperature appears to inhibit directly PEPCase activity, but not the activity of the enzymes responsible for the breakdown of malate, with the result that the leaf acquires a low malate status. In contrast, low temperature does not directly inhibit PEPCase activity, but does inhibit enzymes responsible for malate breakdown, so that the malate level in the leaf increases to a high value and PEPCase is eventually allosterically inhibited. The different malate status of leaves held at these two temperatures accounts for the phases of the rhythms being reversed on returning the leaves to 15°C. After exposure to high temperature, CO₂ fixation by PEPCase activity can begin immediately, whereas after exposure to low temperature, the large amount of malate accumulated in the leaves has to be decarboxylated before CO₂ fixation can begin.     

Enzyme Activities Associated with Carbon Dioxide Exchange in Illuminated Leaves of Hordeum vulgare L.: IV. The Effect of Light Intensity on the Carbon Dioxide Compensation Point

The carbon dioxide compensation point () was found to vary whenmeasured at increasing light intensities, in plants grown ata constant illumination. This response varied with the physiologicalage of the leaf. The also varied when measured at a constantillumination, with plants grown at different light intensities.The activity of the enzymes RuDP carboxylase, nitrate reductase,glycollate oxidase, and catalase was found to be influencedby the light intensity at which the plants were grown. A goodcorrelation was obtained between the measured and the ratioof nitrate reductase: RuDP carboxylase activities, suggestingthat nitrate reductase may be used as an indirect measure ofphotorespiration in plants receiving nitrate as the sole nitrogensource.     

蓝藻浓缩二氧化碳的机制

综述了蓝藻的CO2逍缩机制研究进展,并简要介绍了一些重要的研究手段。     

The Melting Phase Transition in Small Carbon Dioxide Clusters

Abstract

Small isolated carbon dioxide clusters have been studied by canonical and microcanonical Molecular Dynamics simulation. The phenomenon of “dynamical coexistence” between solidlike and liquidlike forms of the (CO₂)₁₃ cluster was found, in a finite energy range. An animation of this phenomenon is shown. Coexistence is associated with bimodality in the probability distribution of the short-time averaged internal kinetic energy of the system in the microcanonical ensemble. This phenomenon has also been observed in (CO₂)_12,14. It is interpreted in terms of the energy distribution of the Potential Energy Surface minima. This statistical mechanics tool enables to understand why dynamical coexistence has also been observed in previous works on (Ar)₁₃ and (N₂)₁₃ but not in the case of (Ar)_8,14 and (SF₆)₁₃.     

Persistent circadian rhythms in the phosphorylation state of phosphoenolpyruvate carboxylase from Bryophyllum fedtschenkoi leaves and in its sensitivity to inhibition by malate

Phosphoenolpyruvate carboxylase (EC 4.1.1.31; PEPCase) from Bryophyllum fedtschenkoi leaves has previously been shown to exist in two forms in vivo. During the night the enzyme is phosphorylated and relatively insensitive to feedback inhibition by malate whereas during the day the enzyme is dephosphorylated and more sensitive to inhibition by malate. These properties of PEPCase have now been investigated in leaves maintained under constant conditions of temperature and lighting. When leaves were maintained in continuous darkness and CO₂-free air at 15°C, PEPCase exhibited a persistent circadian rhythm of interconversion between the two forms. There was a good correlation between periods during which the leaves were fixing respiratory CO₂ and periods during which PEPCase was in the form normally observed at night. When leaves were maintained in continuous light and normal air at 15°C, starting at the end of a night or the end of a day, a circadian rhythm of net uptake of CO₂ was observed. Only when these constant conditions were applied at the end of a day was a circadian rhythm of interconversions between the two forms of PEPCase observed and the rhythms of enzyme interconversion and CO₂ uptake did not correlate in phase or period.Abbreviations CAM Crassulacean acid metabolism - FW fresh weight - PEPCase phosphoenolpyruvate carboxylase - RuBPCase ribulose-1,5-bisphosphate carboxylase To whom correspondence should be addressed.     

A circadian rhythm in the level of carbon dioxide compensation in Bryophyllum fedtschenkoi with zero values during the transient

Summary Detached shoots of Bryophyllum fedtschenkoi maintained in a closed system in the light exhibited an endogenous circadian rhythm in CO₂ compensation. The rhythm was sensitive to changes in light intensity and temperature. At 15° C it damped rapidly in light of 78 J m^-2 s^-1, but at 10° C a rhythm of considerable amplitude was evident at this same light intensity. During the transient (i.e. the temporary state of the rhythm before it acquired its steady state) low compensation values between 0 and 5 ppm CO₂ were achieved. When the plants were maintained at a higher light intensity prior to the measurements, the period of low compensation during the transient was extended, and zero values were obtained under some conditions.Studies of gas exchange at opposite phases of the rhythm revealed: (i) that the rate of uptake of ¹⁴CO₂ differed, both in light and darkness (the epidermis was removed during these observations to avoid interference from stomatal rhythms); (ii) photorespiration, estimated by extrapolation of the graph relating photosynthetic rate and CO₂ concentration, was highest during the peaks of the rhythm in CO₂ compensation; (iii) estimates of the capacity for photorespiration by the glycine-1-¹⁴C assay indicated highest values during the troughs of the rhythm. These findings are discussed in relation to the C₄-acid metabolism of this species. Low CO₂ compensation is probably due to the activity of phosphoenolpyruvate carboxylase and not to the absence of processes involving CO₂ evolution.