Egg release in gametophytes of Laminaria saccharina: Induction by darkness and inhibition by blue light and u.v.

Gametophytes of Laminaria saccharina cultivated from zoospores in a light-dark regime (16:8), release eggs exclusively during the dark cycles, 8–10 days after seeding of the zoospores, and mainly during the first 30 min of darkness. The inhibiting effects of light during the light cycle of 16 h per day is also apparent in gametophytes which have experienced only two dark cycles prior to day 8, when egg release begins. Egg release can be shifted to any time during the light cycle by prolonging the irradiation with white fluorescent light and by subsequent darkening for 1 h. In gametophytes cultivated in continuous white fluorescent light eggs are also released from day 8–10, so in this case no inhibiting activity of light is apparent. Egg release is inhibited by blue light and u.v., with peak wavelengths for inhibition at 372, 413, 438 and 481 nm. No inhibition occurs at wavelengths above 513 nm. The light requirement for inhibition is very low. A photon fluence rate of 1·4 μE m^-2s^-1, given for 45 min at 449 nm, inhibits egg release in 50% of the mature gametophytes. There is some evidence that a circadian rhythm is involved, primarily since in gametophytes which are transferred at the beginning of day 8 from 16:8 to constant conditions (darkness, continuous red or green light) the diel rhythm of egg release persists until day 11.     

Stem extension rate in light-grown plants. Evidence for an endogenous circadian rhythm in Chenopodium rubrum

Stem extension in light-grown plants of Chenopodium rubrum L. ecotype selection 184 (50°10'N; 150°35'W) was recorded continuously for periods up to one week at constant temperature. Stem extension rate measurements were made with linear voltage-displacement transducer devices. At the beginning of experiments, the 3rd intenode above the cotyledons was about 5 mm long. Stem extension rate exhibited a rhythmic behaviour in continuous white light (20 W m⁻²), and in continuous darkness with a period of approximately 23 h. In continuous darkness, the amplitude of the rhythm damped out very quickly after 24 h and a second peak was just measurable. The mean value of the stem extension rate was dependent on the light fluence before the experiments. This overt rhythm, which could be observed at the individual plant or even internode level, exhibited the characteristics of an endogenous circadian rhythm. There was no correlation of the peak time to local time. The peak time was determined by the time of transfer from dark to light for dark periods equal to or longer than 8 h, and the phase was shifted by the time of transfer from light to dark at the proper phase of a pre-existing rhythm.     

The effects of pinealectomy and blinding on the circadian locomotor activity rhythm in the Japanese newt,Cynops pyrrhogaster

We examined the effects of pinealectomy and blinding (bilateral ocular enucleation) on the circadian locomotor activity rhythm in the Japanese newt, Cynops pyrrhogaster. The pinealectomized newts were entrained to a light-dark cycle of 12 h light and 12 h darkness. After transfer to constant darkness they showed residual rhythmicity for at least several days which was gradually disrupted in prolonged constant darkness. Blinded newts were also entrained to a 12 h light/12 h dark cycle. In subsequent constant darkness they showed free-running rhythms of locomotor activity. However, the freerunning periods noticeably increased compared with those observed in the previous period of constant darkness before blinding. In blinded newts entrained to the light/dark cycle the activity rhythms were gradually disrupted after pinealectomy even in the presence of the light/dark cycle. These results suggest that both the pineal and the eyes are involved in the newt's circadian system, and also suggest that the pineal of the newt acts as an extraretinal photoreceptor which mediates the entrainment of the locomotor activity rhythm.Abbreviations circadian period - DD constant darkness - LD cycle, light-dark cycle - LD 12:12 light-dark cycle of 12 h light and 12 h darkness     

Phase resetting of the circadian rhythm of carbon dioxide assimilation inBryophyllum leaves in relation to their malate content following brief exposure to high and low temperatures,darkness and 5% carbon dioxide

Leaves ofBryophyllum fedtschenkoi show a persistent circadian rhythm in CO₂ assimilation when kept in continuous illumination and normal air at 15°C. The induction of phase shifts in this rhythm by exposing the leaves for four hours at different times in the circadian cycle to 40° C, 2° C, darkness and 5% CO₂ have been investigated. Exposure to high temperature has no effect on the phase at the apex of the peak but is effective at all other times in the cycle, whereas exposure to low temperature, darkness or 5% CO₂ is without effect between the peaks and induces a phase shift at all other times. The next peak of the rhythm occurs 17 h after a 40° C treatment and 7–10 h after a 2° C, dark or 5% CO₂ treatment regardless of their position in the cycle. When these treatments are given at times in the cycle when they induce maximum phase shifts, they cause no change in the gross malate status of the leaf. The gross malate content of the leaf in continuous light and normal air at 15% shows a heavily damped circadian oscillation which virtually disappears by the time of the third cycle, but the CO₂ assimilation rhythm persists for many days. The generation of the rhythm, and the control of its phase by environmental factors are discussed in terms of mechanisms that involve the synthesis and metabolism of malate in specific localised pools in the cytoplasm of the leaf cells.     

Synchronization of Mating Reactivity Rhythms in Populations of Paramecium bursaria

Cell populations of Paramecium bursaria show arhythmic mating reactivity after exposure to constant light (LL) for more than 2 wk. After this arhythmic population is exposed to darkness for 9 h, the mating reactivity rhythm of the cell population reappears. The phases of rhythms in individual cells are synchronized to each other. When the arhythmic population in constant light is exposed to dark pulses of various durations, the first peak of the recovered mating reactivity rhythm appears 6 h after the end of the dark pulse. Thus, in the case of dark pulses to cells in LL, the transition from dark to light sets the phase of the subsequent mating reactivity rhythm. When an arhythmic population in LL is transferred to constant darkness (DD), a rhythm of mating reactivity also appears and, in this case, the first peak of the rhythm occurs 18 h after the LL to DD transition. Therefore, arhythmic populations of cells in LL can be synchronized by either a dark pulse or by transition to continuous darkness. When the arhythmic populations in LL were transferred to various light/dark (LD) cycles, the mating reactivity rhythms entrained to LD cycles of 18 to 30 h in duration. Finally, mating rhythms can also be synchronized by treatment with puromycin (400 μg/ml for 6–18 h).     

A Circadian Rhythm in the Number of Daughter Cells in Synchronous Chlorella fusca var vacuolata

Chlorella fusca var vacuolata cells were transferred to continuous darkness or weak light (0.07 watts per square meter) (both were called waiting time, WT) after a 12-hour light and 12-hour dark schedule. A daily dilution is performed at the end of the light/dark schedule, resulting in always the same average production of 18 autospores per mother cell. After 12 and 24 hours of WT in darkness, the production of autospores in a subsequent light/dark schedule was 50 and 100%, respectively. If the WT was performed in weak light (0.07 watts per square meter) the lowest production was obtained, independently of the length of WT. However, an interruption of this weak light by dark pulses (3 hours) increased the autospore production by an amount that depends upon the phase of the circadian rhythm, and varied up to 70% of the control (WT in permanent darkness). If the WT (total darkness) was interrupted by light pulses of 0.5 hour (white, same as used for growth), a phase response curve of productivity resulted. Pulses between the 12th and 18th hour of WT in darkness gave a 3-hour delay of maximum; later on pulses shifted the maximum autospore production 3 hours ahead.     

Melatonin accelerates reentrainment of the circadian rhythm of its own production after an 8-h advance of the light-dark cycle

Summary The rhythm in melatonin production in the rat is driven by a circadian rhythm in the pineal N-acetyltransferase (NAT) activity. Rats adapted to an artificial lighting regime of 12 h of light and 12 h of darkness per day were exposed to an 8-h advance of the light-dark regime accomplished by the shortening of one dark period; the effect of melatonin, triazolam and fluoxetine, together with 5-hydroxytryptophan, on the reentrainment of the NAT rhythm was studied.In control rats, the NAT rhythm was abolished during the first 3 cycles following the advance shift. It reappeared during the 4th cycle; however, the phase relationship between the evening rise in activity and the morning decline was still compressed.Melatonin accelerated the NAT rhythm reentrainment. In rats treated chronically with melatonin at the new dark onset, the rhythm had already reappeared during the 3rd cycle, in the middle of the advanced night, and during the 4th cycle, the phase relationship between the evening onset and the morning decline of the NAT activity was the same as before the advance shift. In rats treated chronically with melatonin at the old dark onset or in those treated with melatonin 8 h, 5 h and 2 h after the new dark onset during the 1st, 2nd and 3rd cycle, respectively, following the advance shift, the NAT rhythm reappeared during the 3rd cycle as well but in the last third of the advanced night only.Neither triazolam nor fluoxetine together with 5-hydroxytryptophan administered around the new dark onset facilitated NAT rhythm reentrainment after the 8-h advance of the light-dark cycle.Abbreviations NAT N-acetyltransferase - LD cycle light-dark cycle - CT circadian time - LD xy light dark cycle comprising x h of light and y h of darkness     

Rhythmic egg-laying behaviour in virgin females of fruit flies Drosophila melanogaster

Fruit fly Drosophila melanogaster females display rhythmic egg-laying under 12:12?h light/dark (LD) cycles which persists with near 24?h periodicity under constant darkness (DD). We have shown previously that persistence of this rhythm does not require the neurons expressing pigment dispersing factor (PDF), thought to be the canonical circadian pacemakers, and proposed that it could be controlled by peripheral clocks or regulated/triggered by the act of mating. We assayed egg-laying behaviour of wild-type Canton S (CS) females under LD, DD and constant light (LL) conditions in three different physiological states; as virgins, as females allowed to mate with males for 1?day and as females allowed to mate for the entire duration of the assay. Here, we report the presence of a circadian rhythm in egg-laying in virgin D. melanogaster females. We also found that egg-laying behaviour of 70 and 90% females from all the three male presence/absence protocols follows circadian rhythmicity under DD and LL, with periods ranging between 18 and 30?h. The egg-laying rhythm of all virgin females synchronized to LD cycles with a peak occurring soon after lights-off. The rhythm in virgins was remarkably robust with maximum number of eggs deposited immediately after lights-off in contrast to mated females which show higher egg-laying during the day. These results suggest that the egg-laying rhythm of D. melanogaster is endogenously driven and is neither regulated nor triggered by the act of mating; instead, the presence of males results in reduction in entrainment to LD cycles.     

Rhythms as photoperiodic timers in the control of flowring in Chenopodium rubrum L.

Summary In C. rubrum, the amount of flowering that is induced by a single dark period interrupting continuous light depends upon the duration of darkness. A rhythmic oscillation in sensitivity to the time that light terminates darkness regulates the level of flowering. The period length of this oscillation is close to 30 hours, peaks of the rhythm occurring at about 13, 43 and 73 h of darkness.Phasing of the rhythm by 6-, 12- and 18-h photoperiods was studied by exposing plants to a given photoperiod at different phases of the free-running oscillation in darkness. The shift in phase of the rhythm was then determined by varying the length of the dark period following the photoperiod; this dark period was terminated by continuous light.With a 6-h photoperiod the timing of both the light-on and light-off signals is shown to control rhythm phasing. However, when the photoperiod is increased to 12 or 18 h, only the light-off signal determines phasing of the rhythm. In prolonged periods of irradiation-12 to 62 h light—a durational response to light overrides any interaction between the timing of the light period and the position of the oscillation at which light is administered. Such prolonged periods of irradiation apparently suspend or otherwise interact with the rhythm so that, in a following dark period, it is reinitiated at a fixed phase relative to the time of the light-off signal to give a peak of the rhythm 13 h after the dusk signal.In daily photoperiodic cycles rhythm phasing by a 6-h photocycle was also estimated by progressively increasing the number of cycles given prior to a single dark period of varied duration.In confirmation of Bünning's (1936) hypothesis, calculated and observed phasing of the rhythm controlling flowering in c. rubrum accounts for the photoperiodic response of this species. Evidence is also discussed which indicates that the timing of disappearance of phytochrome P_fr may limit flowering over the early hours of darkness.     

Circadian rhythms in the mating behavior of the cockroach, Leucophaea maderae

Mating behavior of small populations of virgin males and females of the cockroach Leucophaea maderae were continuously monitored via time-lapse video recording in controlled laboratory conditions. The time of onset of copulation was found to be rhythmic in a light cycle of 12 h light alternated with 12 h of darkness, with the peak of mating behavior occurring near the light to dark transition. This rhythm persisted in constant dim red illumination and constant temperature. In constant conditions, the period of the rhythm was slightly less than 24 h, with a peak of copulation during the late subjective day. These data demonstrated that mating behavior is gated by a circadian clock. When males and females were taken from light cycles that were 12 h out of phase, a bimodal rhythm was observed with one peak in the males' late subjective day and a second peak of equal amplitude in the late subjective day of females. The results indicated that circadian systems in both males and females contribute to the circadian rhythm in copulation. Bilateral section of the optic tracts (OTX) of both males and females abolished the rhythm, but the rhythm persisted when OTX females were paired with intact males or when OTX males were paired with intact females. Furthermore, when OTX males or OTX females were paired with intact animals that were 12 h out of phase, a bimodal rhythm was still observed. These results suggested that the circadian pacemaker in the optic lobes of both male and female cockroaches participates in the control of mating, but that a pacemaker outside the optic lobes is also likely involved. Finally, it was shown that the female's olfactory response (measured by electroantennogram) to components of the male sex pheromone exhibited a circadian rhythm, but the data suggested the peripheral olfactory rhythm is not likely to be involved in the rhythm of mating behavior.     

Evidence of phytochrome involvement in the entrainment of the circadian rhythm of carbon dioxide metabolism in Bryophyllum

The rhythm of carbon dioxide output in Bryophyllum leaves was entrained on exposure to 0.25 h of white light every 24 h. Entrainment also occurred on similar exposure to monochromatic radiation in spectral bands centred at 660 nm and, to a lesser extent, at 730 nm, but a band centred at 450 nm was without effect. A skeleton irradiation programme comprising two 0.25-h exposures to white light per 24 h also entrained the rhythm when the intervening dark periods were either 7.5 h and 16 h, or 10.5 h and 13 h. The rhythm disappeared when the two exposures were separated by 11.5-h and 12-h dark periods. Regular 0.25-h exposures to red light separated by 11.75-h periods of darkness also resulted in loss of the rhythm. Red/far-red reversibility was observed in irradiation schedules having either one or two exposures to red light daily. In the latter case, far-red reversal of the effects of one of the exposures to red light resulted in entrainment of the rhythm by the other, instead of abolition of the rhythm. The occurrence of distinct red/far-red reversibility suggests strongly that phytochrome is the pigment involved in entrainment of this rhythm by cycles of light and darkness.Abbreviation LD light-dark rhythm     

Regulation of PEP-Carboxylase by Biological Clock in a CAM Plant

The endogenous circadian rhythm in a crassulacean acid metabolism(CAM) plant Graptopetalum paraguayense was investigated. Phosphoenolpyruvatecarboxylase (PEP-C) takes two forms: the malate-sensitive dayform and the malate-insensitive night form. We monitored thestate of PEP-C by measuring the sensitivity to malate as a parameterof the circadian rhythm. We also measured vacuolar pH and malateconcentration, and contents of oxaloacetate, pyruvate and phosphoenolpyruvate(PEP). A free-running circadian oscillation was observed under continuousdim light (5 klux) after 12 h/12 h light/dark cycles at 20°C.The period of the rhythm was about 20 h. Under continuous light(18 klux), the rhythm was less clear but the length of the periodwas not affected. On the other hand, the rhythms of the vacuolarpH and the malate concentration were evident under the continuouslight, but were not clear under the continuous dim light. Therhythm disappeared in continuous darkness. The content of PEPchanged simultaneously with the transformation of PEP-C duringthe normal day-night cycles and under the continuous light,but stayed at a low level under the continuous dim light. Thisindicated that the transformation of PEP-C was not sufficientto maintain the rhythm in the carbon metabolism. Shift of the timing of the start or end of the dark period priorto the continuous illumination shifted the phase of the PEP-Crhythm without changing the period length significantly. At30°C, the rhythm of PEP-C was less clear, but the periodlength was not affected. These results suggest that the biological clock controls CO₂uptake and day-night CAM cycle through regulation of PEP-C transformation. (Received August 20, 1993; Accepted December 3, 1993)     

PRESENCE OF CIRCADIAN RHYTHMS IN THE LOCOMOTOR ACTIVITY OF A CAVE-DWELLING MILLIPEDE GLYPHIULUS CAVERNICOLUS SULU (CAMBALIDAE,SPIROSTREPTIDA)

The locomotor activity of the millipede Glyphiulus cavernicolus (Spirostreptida), which occupies the deeper recesses of a cave, was monitored in light-dark (LD) cycles (12h light and 12h darkness), constant darkness (DD), and constant light (LL) conditions. These millipedes live inside the cave and are apparently never exposed to any periodic factors of the environment such as light-dark, temperature, and humidity cycles. The activity of a considerable fraction of these millipedes was found to show circadian rhythm, which entrained to a 12:12 LD cycle with maximum activity during the dark phase of the LD cycle. Under constant darkness (DD), 56.5% of the millipedes (n = 23) showed circadian rhythms, with average free-running period of 25.7h ± 3.3h (mean ± SD, range 22.3h to 35.0h). The remaining 43.5% of the millipedes, however, did not show any clear-cut rhythm. Under DD conditions following an exposure to LD cycles, 66.7% (n = 9) showed faint circadian rhythm, with average free-running period of 24.0h ± 0.8h (mean ± SD, range 22.9h to 25.2h). Under constant light (LL) conditions, only 2 millipedes of 11 showed free-running rhythms, with average period length of 33.3h ± 1.3h. The results suggest that these cave-dwelling millipedes still possess the capacity to measure time and respond to light and dark situations. (Chronobiology International, 17(6), 757–765, 2000)     

An investigation of circadian rhythm in Escherichia coli

Background: Persuasive evidence for circadian programs in non-photosynthetic bacteria other than cyanobacteria is still lacking, we aimed to investigate the circadian rhythm of specific growth rate in Escherichia coli ATCC 25922, one of the important prokaryotes. Methods: To grow E. coli under different light and dark conditions. When the growth entered into the stationary phase, we stopped the culture and obtained the viable counts by MTT assay every 3 h. The specific growth rates (SGRs) were calculated and analyzed with cosinor method for potential rhythms. Results: Single cosinor method revealed that the SGR of E. coli displayed rhythmic variations with a period of around 24 h both under light/dark cycles and under constant darkness. The best-fitting periods and best-fitting cosine curves were acquired. Conclusions: The SGR of E. coli (ATCC 25922) in a culture medium with limiting substrates in the stationary and death phases displayed rhythmic variations with a period of around 24 h under light/dark cycles and constant darkness conditions.     

Studies on Halimeda IV. An endogenous rhythm of chloroplast migration in the siphonous green alga,H. distorta 1

Abstract

Halimeda has been found particularly suitable for studies of long‐distance chloroplast migration by virtue of its coenocytic structure and calcium carbonate skeleton. A circadian rhythm of chloroplast migration in Halimeda distorta was monitored by videography of segment surface pigmentation. In normal 12 h light/12 h dark treatments synchronised with dawn and dusk, the segments were green all day, began to become pale immediately the light was turned off, and then remained almost white for most of the night until beginning to re‐green a few hours before dawn. As a result of that, they were already quite green by the time the light was turned on. In continuous darkness a similar cycle, albeit with reducing amplitude and a period of about 23 hours, was maintained for at least 7 days. However, this cycle differed significantly from the normal one in that the segments did not remain green after the light was not switched on at dawn, but rather began to pale immediately thereafter. Conversely, in continuous light the segments did not become pale at any time. Thus, the rhythmical re‐emergence of the chloroplasts before dawn and their subsequent withdrawal appears to be controlled by an endogenous rhythm which is independent of light. However, light does completely, but reversibly, inhibit the chloroplast withdrawal component of the cycle. This behaviour of the chloroplasts in Halimeda is very similar to that in the related alga, Caulerpa, but it is quite different from that in another extensively Studied but unrelated siphonous green alga, Acetabularia, in which the circadian rhythm of chloroplast migration is maintained in continuous light.     

A circadian rhythm influencing foraging behaviour in the saw-toothed grain beetle Oryzaephilus surinamensis

ABSTRACT. Adults of Oryzaephilus surinamensis (L.) (Coleoptera, Silvanidae) placed in an open arena containing a refuge showed a cyclic pattern of activity in light, dark (LD) cycles when food in the form of damaged wheat grains was placed in the arena outside the refuge. Placing food in the refuge reduced cyclic change and lowered the general activity of the beetles. Batches of fifty insects conditioned in LD 9:15 h or 15:9 h at 25°C , 65% r.h., and transferred to continuous darkness (DD) at the end of a photophase, showed a circadian rhythm of foraging activity of periodicity near 24 h. Those transferred to continuous light at the end of a scotophase showed a 6 h delay in the onset of the next peak of activity, but subsequent peaks, although damped, revealed a periodicity near that in DD. The mean number of beetles wandering in the arena ranged from about eight in LD 15:9 h with all food in the refuge to about twenty-one in LD 9:15 h with all food in the arena.     

Circadian Rhythms in Stomatal Responsiveness to Red and Blue Light

Stomata of many plants have circadian rhythms in responsiveness to environmental cues as well as circadian rhythms in aperture. Stomatal responses to red light and blue light are mediated by photosynthetic photoreceptors; responses to blue light are additionally controlled by a specific blue-light photoreceptor. This paper describes circadian rhythmic aspects of stomatal responsiveness to red and blue light in Vicia faba. Plants were exposed to a repeated light:dark regime of 1.5:2.5 h for a total of 48 h, and because the plants could not entrain to this short light:dark cycle, circadian rhythms were able to "free run" as if in continuous light. The rhythm in the stomatal conductance established during the 1.5-h light periods was caused both by a rhythm in sensitivity to light and by a rhythm in the stomatal conductance established during the preceding 2.5-h dark periods. Both rhythms peaked during the middle of the subjective day. Although the stomatal response to blue light is greater than the response to red light at all times of day, there was no discernible difference in period, phase, or amplitude of the rhythm in sensitivity to the two light qualities. We observed no circadian rhythmicity in net carbon assimilation with the 1.5:2.5 h light regime for either red or blue light. In continuous white light, small rhythmic changes in photosynthetic assimilation were observed, but at relatively high light levels, and these appeared to be attributable largely to changes in internal CO2 availability governed by stomatal conductance.     

Light Cycles Given During Development Affect Freerunning Period of Circadian Locomotor Rhythm of period Mutants in Drosophila melanogaster

We reared wild type (Canton-S) and period mutant flies, i.e., per(S) and per(L), of Drosophila melanogaster in constant darkness, constant light or 24h light dark cycles with various light to dark ratios throughout the development from embryo to early adult. The locomotor activity rhythms of newly eclosed individuals were subsequently monitored in the lighting conditions, in which they had been reared, for several days and then in constant darkness. Circadian rhythms were clearly exhibited in constant darkness even in flies reared in constant light and constant darkness as well as flies reared in light-dark cycles, but the freerunning period differed among groups. The results suggest that the circadian clock is assembled without any cyclical photic information, and that the light influences the developing circadian clock of Drosophila to alter the freerunning period. The effects of light on the rhythm differed in some aspects between per(L) flies and the other two strains. Possible mechanisms through which light affects the developing circadian clock are discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved