Circadian Modulation of Circumnutation Length, Period, and Shape in Helianthus annuus

Modulation of a turgor-growth movement called circumnutation in sunflower (Helianthus annuus L.) was investigated using a picture analysis system. Two photoperiod conditions were applied: light–darkness conditions (LD) 8:8 and LD 20:10. After about 3 weeks of these regimes, the plants were placed under constant light to determine whether circadian regulation of circumnutation existed or not. The rhythms of movement activity with regard to the trajectory length, period, and shape of individual circumnutations were examined. Data were processed by Fourier spectral analysis. All the parameters, trajectory length, period, and shape, revealed the ability to entrain to the administered daily cycles (16 h or 30 h). We observed diurnal fluctuations of the circumnutation parameters with the phase of the highest trajectory length, the shortest period, and the highest shape coefficient (the most circular form) during the dark period. After the LD–LL transition, the parameters revealed periodicity, which was close to 24 h. After several days of a clear circadian free running rhythm, a gradual decrease of the amplitude of the rhythm was observed. However, the rhythm did not disappear completely. The trajectory length manifested the strongest entrainment; the circumnutation period and the circumnutation shape were less modulated by photoperiod. These findings indicate for the first time that different parameters of circumnutation in sunflower are circadian-regulated rhythms, not solely ultradian as had been thought previously.     

Circadian rhythm of circumnutation in inflorescence stems of Arabidopsis

We investigate the modulation of circumnutation in inflorescence stems of Arabidopsis to determine the circadian regulation of circumnutation. Under constant light conditions (LL), circumnutation speed in wild-type plants fluctuates, with the phase of the highest speed at subjective dawn; the period length is close to 24 h. toc1 appears to shorten the period and elf3 causes an arrhythmic phenotype in circumnutation speed in LL, suggesting that a common circadian clock may control both circumnutation speed and other circadian outputs. These results highlight for the first time a role for a circadian clock in the regulation of circumnutation based on genetic analysis of Arabidopsis.     

Complex relationship between growth and circumnutations in Helianthus annuus stem

The growth and circumnutation of the stem of three-week old Helianthus annuus in the 16:8 h light:dark photoperiod were monitored using an angular position-sensing transducer and a time lapse photography system. It was found that the rate of growth and circumnutation reached a high level in the dark stage; in the light stage, however, only the growth rate reached the same high level, whereas the circumnutations were weak. These results showed that in the light stage the stem circumnutation was downregulated more strongly than the growth. Short-term stem responses to darkening and illumination were a further display of the relation between growth and circumnutations. Switching off the light caused an increase in the growth and circumnutation rate. In some cases it was accompanied by changes in the rotation direction. On the other hand, switching the light on caused an immediate transient (several-minute long) decrease in the growth rate resulting in stem contraction, and this was accompanied by an almost complete pause of circumnutation. Additionally, under light, there occurred a subsequent decrease in the magnitude, disturbance of circumnutation trajectory and, in some cases, changes in the direction of rotation. The observed stem contraction and disturbance of circumnutation imply the occurrence of turgor changes in sunflower stem, which may be caused by a non-wounding, darkening or illumination stimulus. Our experiments indicate that the disturbances of the growth rate are accompanied by changes in circumnutation parameters but we have also seen that there is no simple quantitative relation between growth rate and circumnutation rate.Key words: Helianthus annuus, plant movement, circumnutation, elongation, growth, stem contraction     

Circumnutation as a visible plant action and reaction: Physiological,cellular and molecular basis for circumnutations

Circumnutation is a helical organ movement widespread among plants. It is variable due to a different magnitude of trajectory (amplitude) outlined by the organ tip, duration of one cycle (period), circular, elliptical, pendulum-like or irregular shape and clock- and counterclockwise direction of rotation. Some of those movement parameters are regulated by circadian clock and show daily and infradian rhythms. Circumnutation is influenced by light, temperature, chemicals and can depend on organ morphology. The diversity of this phenomenon is easier to see now that the digital time-lapse video method is developing fast. Whether circumnutation is an endogenous action, a reaction to exogenous stimuli or has a combined character has been discussed for a long time. Similarly, the relationship between growth and circumnutation is still unclear. In the mechanism of circumnutation, epidermal and endodermal cells as well as plasmodesmata, plasma membrane, ions (Ca²⁺, K⁺ and Cl⁻), ion channels and the proton pump (H⁺ATPase) are engaged. Based on these data, the hypothetical electrophysiological model of the circumnutation mechanism has been proposed here. In the recent circumnutation studies, gravitropic, auxin, clock and phytochrome mutants are used and new functions of circumnutation in plants'' life have been investigated and described.Key words: circumnutation, Helianthus annuus, plant movement, elongation, growth, ultradian rhythm, circadian rhythm, time-lapse video, ion channels     

Circumnutations under free-fall conditions in space?

Regular growth movement, so-called circumnutation, of plants and plant organs is a very general phenomenon. The origin of circumnutations is, however, unclear. Three different types of model are discussed in relation to some experimental results on circumnutations. It is suggested that an experiment in space, under free-fall conditions, should be done on different species showing circumnutations in order to discriminate between the models.     

Circumnutations of Arabidopsis thaliana Seedlings

The hypocotyls of Arabidopsis thaliana seedlings exhibit circumnutations with different frequency ranges. Using a picture-analysis system, two types of oscillations were distinguished, short period (SPN) and long period nutations (LPN). The period of the SPNs is between 20 and 60 min, decreases with increasing temperature (between 20° and 30°C; Q 10 of 2.0) and increases with age. The SPNs changed frequently between circular and pendular movements. The circumnutation usually occurred clockwise, but the direction could change. For SPNs to occur, a minimum growth rate of 0.05 mm/h was required. The period of the LPNs ranged from 1 to 8 h. Peaks occurred around 95 and 200 min. The movements of the LPNs are mostly counterclockwise. The oscillation could change from circular to elliptic or pendular. In contrast to SPNs, LPNs were found also at low growth rates. Under red light and in the strain Landsberg erecta the circumnutations were especially slow. Short and long period oscillations can occur simultaneously or separately in different plants. In cases of simultaneous occurrence the quotient between the period lengths of the LPN and SPN was not constant. The number of occurrences of these frequencies depended on the strain and on external factors such as temperature and light. In continuous bright white light phases of circumnutations alternated with quiescent periods. This behaviour was circadian and correlated with growth bursts.     

Circumnutations of Arabidopsis thaliana Seedlings

The hypocotyls of Arabidopsis thaliana seedlings exhibit circumnutations with different frequency ranges. Using a picture-analysis system, two types of oscillations were distinguished, short period (SPN) and long period nutations (LPN). The period of the SPNs is between 20 and 60 min, decreases with increasing temperature (between 20° and 30°C; Q 10 of 2.0) and increases with age. The SPNs changed frequently between circular and pendular movements. The circumnutation usually occurred clockwise, but the direction could change. For SPNs to occur, a minimum growth rate of 0.05 mm/h was required. The period of the LPNs ranged from 1 to 8 h. Peaks occurred around 95 and 200 min. The movements of the LPNs are mostly counterclockwise. The oscillation could change from circular to elliptic or pendular. In contrast to SPNs, LPNs were found also at low growth rates. Under red light and in the strain Landsberg erecta the circumnutations were especially slow. Short and long period oscillations can occur simultaneously or separately in different plants. In cases of simultaneous occurrence the quotient between the period lengths of the LPN and SPN was not constant. The number of occurrences of these frequencies depended on the strain and on external factors such as temperature and light. In continuous bright white light phases of circumnutations alternated with quiescent periods. This behaviour was circadian and correlated with growth bursts.     

Circumnutations of Sunflower Hypocotyls in Satellite Orbit

The principal objective of the research reported here was to determine whether a plant's periodic growth oscillations, called circumnutations, would persist in the absence of a significant gravitational or inertial force. The definitive experiment was made possible by access to the condition of protracted near weightlessness in an earth satellite. The experiment, performed during the first flight of Spacelab on the National Aeronautics and Space Administration shuttle, Columbia, in November and December, 1983, tested a biophysical model, proposed in 1967, that might account for circumnutation as a gravity-dependent growth response. However, circumnutations were observed in microgravity. They continued for many hours without stimulation by a significant g-force. Therefore, neither a gravitational nor an inertial g-force was an absolute requirement for initation or continuation of circumnutation. On average, circumnutation was significantly more vigorous in satellite orbit than on earth-based clinostats. Therefore, at least for sunflower (Helianthus annuus L.) circumnutation, clinostatting is not the functional equivalent of weightlessness.     

Influence of the g-force on the circumnutations of sunflower hypocotyls

Circumnutations of hypocotyls of sunflower ( Helianthus annuus L. cv. Californicus) were studied under 1 g and 3 g conditions. Root mean square values of the hypocotyl deviation from the plumbline and period of the movements were determined from Calculations of the autocorrelation functions of the movements. The amplitude and the period of the circumnutations increased under 3 g as compared to 1 g . A transition from 3 to 1 g or vice versa also caused changes in period and amplitude of the movoments. The results are interpreted as a support for the idea that gravity influences the circumnutation parameters in this sunflower variety. A comparison is made with published results on the dwarf sunflower ev. Teddy Bear where the force influence is very small or negligible. Simulations of a model for circumnutations show movements which are in qualitative agreement with the experimental results, provided adaptation to g -levels is included in the model. Finally, the results are discussed with the recent Spacelab-experiment (SLI) as a background.     

KINETICS OF SUPPRESSION OF CIRCUMNUTATION BY CLINOSTATTING FAVORS MODIFIED INTERNAL OSCILLATOR MODEL

According to a well-conceived gravity-driven model, oscillatory growth movements called circumnutations are produced by a continuing set of gravitational responses with overshoot. In simplest form that model predicts that if the gravity stimulus could be removed completely and abruptly, growth oscillations would damp out within a time less than the period of oscillation. That condition was simulated by imposing on horizontally clinostatted sunflower shoots a unit g centripetal force in the direction that coincided with the plants' morphological axis. Circumnutations proceeded in quantitatively normal manner. By stopping the centrifuge, the plants were subjected abruptly only to clinostat simulated weightlessness. Circumnutations decreased but did not disappear. The half-time for decline to a final low level of activity was about 4-fold the period of oscillation at 1 g—about 8-fold the period at simulated zero g. That slow damping was not consistent with the prediction based on the gravity driven model of circumnutation. The model needs to be revised.     

Spontaneous action potentials and circumnutation in <Emphasis Type="Italic">Helianthus annuus</Emphasis>

Action potentials (APs) in plants are involved in fast leaf or trap closure as well as elongation, respiration, photosynthesis, and fertilization regulation. Here, spontaneous APs (SAPs) in relation to endogenous stem movement named circumnutation (CN) have been investigated in Helianthus annuus in different light conditions in freely circumnutating and immobilized plants. Extracellular electrical measurements and time-lapse photography were carried out simultaneously. The parameters of CN (trajectory length, period, and direction) and the number and transmission direction of SAPs were analysed. In continuous light (25–40 μmol m^?2 s^?1), all plants circumnutating vigorously in a regular elliptical manner and no SAPs were observed. In light/dark conditions, the plants circumnutated in a daily pattern, most SAPs were observed in the dark and freely circumnutated sunflowers had two times more SAPs (10 SAPs/24 h/plant) than the immobilized plants (5 SAPs/24 h/plant). In continuous very low light (5 μmol m^?2 s^?1), the plants circumnutated weakly and irregularly and SAPs appeared without the circadian pattern. 3–5 SAPs/24/plant occurred in the freely circumnutating and immobilized plants. In light/dark and continuous very low light conditions, an ultradian rhythm of SAPs was observed and the mean spacing between SAPs was approx. 121–271 min in the freely circumnutating and immobilized plants. Under all light conditions, more SAPs were transmitted basipetally than acropetally. One-hour lasting series of 3–4 min spaced SAPs locally propagated were observed as well in very low light. Basipetal and acropetal SAPs passing along the stem motor region accompany irregularity, changes in the CN trajectory direction, and stem torsion. These results demonstrate that APs and CN changes play a role in plant adaptation to light conditions and that there is an ultradian rhythm of SAPs beside ultradian CN rhythm.     

Effects of constant darkness and constant light on circadian organization and reproductive responses in the ram

The relationship between circadian rhythms in the blood plasma concentrations of melatonin and rhythms in locomotor activity was studied in adult male sheep (Soay rams) exposed to 16-week periods of short days (8 hr of light and 16 hr of darkness; LD 8:16) or long days (LD 16:8) followed by 16-week periods of constant darkness (dim red light; DD) or constant light (LL). Under both LD 8:16 and LD 16:8, there was a clearly defined 24-hr rhythm in plasma concentrations of melatonin, with high levels throughout the dark phase. Periodogram analysis revealed a 24-hr rhythm in locomotor activity under LD 8:16 and LD 16:8. The main bouts of activity occurred during the light phase. A change from LD 8:16 to LD 16:8 resulted in a decrease in the duration of elevated melatonin secretion (melatonin peak) and an increase in the duration of activity corresponding to the changes in the ratio of light to darkness. In all rams, a significant circadian rhythm of activity persisted over the first 2 weeks following transfer from an entraining photoperiod to DD, with a mean period of 23.77 hr. However, the activity rhythms subsequently became disorganized, as did the 24-hr melatonin rhythms. The introduction of a 1-hr light pulse every 24 hr (LD 1:23) for 2 weeks after 8 weeks under DD reinduced a rhythm in both melatonin secretion and activity: the end of the 1-hr light period acted as the dusk signal, producing a normal temporal association of the two rhythms. Under LL, the 24-hr melatonin rhythms were disrupted, though several rams still showed periods of elevated melatonin secretion. Significant activity rhythms were either absent or a weak component occurred with a period of 24 hr. The introduction of a 1-hr dark period every 24 hr for 2 weeks after 8 weeks under LL (LD 23:1) failed to induce or entrain rhythms in either of the parameters. The occurrence of 24-hr activity rhythm in some rams under LL may indicate nonphotoperiodic entrainment signals in our experimental facility. Reproductive responses to the changes in photoperiod were also monitored. After pretreatment with LD 8:16, the rams were sexually active; exposure to LD 16:8, DD, or LL resulted in a decline in all measures of reproductive function. The decline was slower under DD than LD 16:8 or LL.(ABSTRACT TRUNCATED AT 400 WORDS)     

Circadian rhythms of demand-feeding and locomotor activity in rainbow trout

Under free-running conditions, most rainbow trout displayed circadian feeding rhythms, although the expression of circadian rhythmicity depended on the experimental condition: 16·7% of fish under constant dim light (LL dim), 66·1% under a 45 :45 min light-dark cycle (LD pulses), and 83·8% under constant light (LL). Under LD pulses, the period length of the free-running rhythms for feeding was significantly shorter (21·9 ± 0·7 h, n =8) than under LL (26·2 ± 0·3 h, n =10). Period length for locomotor activity under LL was 25·8 ± 0·6 h ( n =4). Under LD conditions, the daily demand-feeding profile was always confined to the light phase and chiefly composed of two main episodes, directly after lights on (light elicited) and in anticipation to lights off (endogenous). Contrasting to feeding, the diel locomotor activity profile varied remarkably: a diurnal activity pattern at the bottom, while a clearly nocturnal pattern at the surface. These results contribute to a better understanding of feeding and locomotor rhythms of rainbow trout, providing evidence for the existence of a biological clock involved in their circadian control. This finding contrasts with the previously recorded lack of an endogenous oscillator in the pineal organ driving the rhythmic secretion of melatonin, which suggests different locations from the pineal for the circadian pacemakers in this species.     

The internal synchronization of five circadian functions of the rabbit

Five different physiological functions of the rabbit (hard faeces and urine excretion, food and water intake and locomotor activity) were registered during LD 12:12 and during continuous light conditions (LL).

(1) In LD 12:12 a strong synchronization of the five parameters existed. The minima of all functions consistently occurred during the hours of light. The nocturnal percentage of overall 24-hr events was increased significantly in 'hard faeces excretion' (66±8 (S.D.) %), 'water intake' (64±15 (S.D.) %) and 'urine excretion' (58±10 (S.D.) %). The nocturnal percentage of locomotor activity was significantly increased during the dark-hours in 9 out of 14 animals. In the other five individuals prominent peaks were present even during the photoperiod. On the average of all 14 animals 5S±13 (S.D.) % of the 24 hr events of locomotor activity occurred during the night. Despite a trough during the cessation of hard faeces excretion the events of food intake were not elevated significantly during the dark hours.

(2) During LL the synchronization of the five functions within each animal persisted during the complete 90-day LL period. A free-running circadian rhythm with-: = 24.8±0.5 (S.D.) hr was present in the four rabbits kept in LL conditions within 5-16 days after the withdrawal of the zeitgeber.

(3) In addition to the circadian period the power spectrum analysis of data obtained during LD 12:12 revealed significant ultradian periods of an average period length of 11,6 hr (hard faeces and urine excretion), 8 hr (food and water intake, locomotor activity) and 4 hr (food intake, locomotor activity). During the free-run ultradian periods of 8 and 3.2-4.2 hr were significant in almost all parameters.

(4) During LL the level of locomotor activity was reduced for 13±16 (S.D.) %, the events of food intake were increased for 17±12 (S.D.) %.

(5) The reinserted LD 12:12 zeitgeber re-entrained the circadian rhythms within 25-45 days.

(6) These results provided evidence of a predominant nocturnality of the rabbits under investigation.     

Oscillations in the Activities of Enzymes of Nitrate Reduction and Ammonia Assimilation in Glycine max and Zea mays

The activities of glutamate dehydrogenase (GDH), glutamine synthetase (GS), and nitrate reductase (NR) and the levels of soluble protein and NO^-₃ were assayed in soybean (Glycine max [L.] Merr.) leaves over a 48-h period with the initial 24 h under a light-dark cycle (LD 16:8) followed by 24 h of continuous light (LL). Plants had been entrained for 30 days under the LD regime. Maize (Zea mays) leaves (10 days old) under a LD 15:9 cycle were assayed only for NR and nitrite reductase (NiR). Data were subjected to frequency analysis by the least squares method to determine probabilities for cosine function periods (τ's) between 10 and 30 h. NR activities for both soybean and Zea leaves had 24 h τ's with P values < 0.05 indicating circadian periodicity. GDH in soybeans had a 24-h rhythm under LD conditions which lengthened under LL conditions. The 24-h rhythm of GDH displayed maximal activity toward the end of the dark period of the LD cycle whereas the highest activity of NR was early in the light period. Total soluble protein displayed a rhythm with a best fitting τ of greater than 24 h under both LD and LL. GDH, GS, NR, NO₃, and soluble protein in soybeans and NiR in Zea, all displayed that were ultradian (10–18 h), indicating that a τ of about one half a circadian periodicity may be a common characteristic of the enzymes of primary nitrogen metabolism in higher plants. These data also demonstrate that although both NR and GDH are circadian in their activity, the 24-h rhythm may be greatly influenced by ultradian oscillations in activity.     

Some effects of photoperiod on larviposition and fresh weight-gain in Myzus persicae

ABSTRACT. The larviposition of adult apterous Myzus persicae previously entrained to LD 18:6 showed a marked rhythm both under LD 12:12 (on plants reared in LD 18:6) and under LL (on plants reared in LL). No rhythm was detectable in larviposition by adults reared in LL. Larviposition peaked towards the end of the photophase in LD 12:12. Fresh-weight gain also showed a circadian rhythm with the greatest weight increases during the photophase. Re-entrainment from LD 12:12 to DL 12:12 was not complete after 10 days. It is concluded that the changes in the light cycle did not affect the aphids through the plant.     

Daily rhythms of clock gene expression and feeding behavior during the larval development in gilthead seabream,Sparus aurata

Light is the main environmental time cue which synchronizes daily rhythms and the molecular clock of vertebrates. Indeed, alterations in photoperiod have profound physiological effects in fish (e.g. reproduction and early development). In order to identify the changes in clock genes expression in gilthead seabream larvae during ontogeny, three different photoperiods were tested: a regular 12L:12D cycle (LD), a continuous light 24L:0D (LL) and a two-phases photoperiod (LL?+?LD) in which the photoperiod changed from LL to LD on day 15 after hatching (dph). Larvae were sampled on 10, 18, 30 and 60 days post-hatch (dph) during a 24?h cycle. In addition to the expression of clock genes (clock, bmal1, cry1 and per3), food intake was measured. Under LD photoperiod, larvae feed intake and clock genes expression showed a rhythmic pattern with a strong light synchronization, with the acrophases occurring at the same hour in all tested ages. Under LL photoperiod, the larvae also showed a rhythmic pattern but the acrophases occurred at different times depending on the age, although at the end of the experiment (60 dph) clock genes expression and feed intake rhythms were similar to those larvae exposed to LD photoperiod. Moreover, the expression levels of bmal1 and cry1 were much lower than in LD photoperiod. Under the LL?+?LD photoperiod, the 10 dph larvae showed the same patterns as LL treatment while 18 and 30 dph larvae showed the same patterns as LD treatment. These results revealed the presence of internal factors driving rhythmic physiological responses during larvae development under constant environmental conditions. The LL?+?LD treatment demonstrates the plasticity of the clock genes expression and the strong effect of light as synchronizer in developing fish larvae.     

Rhythmic Circumnutation Movement of the Shoots in Phaseolus Vulgaris L

The apex of growing stems in twining plants describes a rhythmic movement in space called circumnutation. By the method of orthogonal projections, the position in space of the apex can be determined constantly. The mathematical analysis of data allowed us to determine that far Phaseolus vulgaris L. (cv. Mangetout Blanc de Juillet) grown under constant illumination and temperature (25°C) the period is about 100 min. This movement has been related to rhythmic changes in the osmotic potential of the cells located in the bending zone of the stem for a constant period. These variations are longitudinally and laterally coordinated. Treatment with a solution of LiCI at 7 × 10 'M supplied to the root system induces a lengthening of the period. The effect of the treatment is reversed by K⁺ ions. From these results we deduce that rhythmic changes of the membranes are implicated in the circumnutation movements of twining plants.