The Influence of Light Wavelength on Phase-Dependent Responsiveness of the Photoperiodic Clock in Migratory Blackheaded Bunting

We investigated the phase-dependent effects of light wavelength on photoperiodic clock in the migratory blackheaded bunting. Two experiments were performed, employing a skeleton paradigm (6 hours light : 6 hours darkness : 1 hour light : 11 hours darkness; 6L : 6D : 1L : 11D) at 37 ± 2 lux intensity. In the experiment 1, both 6 and 1 h light pulses were given at the same wavelength, 500 nm (green) or 650 nm (red). A group exposed to both pulses of white light served as control. In the experi-ment 2, the two light pulses were given at two different wavelengths, 6 h at 500 nm (green) and 1 h at 640 nm (red) in one group or vice-versa in the other. There was almost no photoinduction when both light pulses in experiment 1, or 1 h light pulse in experiment 2, were green. On the other hand, birds fattened and testes recrudesced when both the light pulses in experiment 1, or 1 h light pulse in experiment 2, were red. Birds receiving both pulses of white light in experiment 1 showed an intermediate response. Taken together, these results indicate that the photoperiodic clock in buntings is differentially responsive at its various circadian phases to different light wavelengths.     

Wavelength dependency of light-induced effects on photoperiodic clock in the migratory blackheaded bunting (Emberiza melanocephala)

The effects of light wavelength on photoperiodic clock were determined in the migratory male blackheaded bunting (Emberiza melanocephala). We constructed an action spectrum for photoperiodic induction (body fattening, gain in body mass, and gonadal recrudescence) by exposing birds for 4.5 weeks to 13 h light per day (L:D = 13:11 h) of white (control), blue (450 nm), or red (640 nm) color at irradiances ranging from 0.028 to 1.4Wm(-2). The threshold light irradiance for photoinduction was about 10-fold higher for blue, compared to red and white light. Phase-dependent effects of light wavelength on the photoperiodic clock were further examined in the next two sets of skeleton photoperiods (SKPs). In the first set of SKPs, birds were exposed for four weeks to asymmetrical light periods (L:D:L:D= 6:6:1:11 h) at 0.25+/-0.01 W m(-2); two light periods applied were of the same (450nm: blue:blue, B:B; 640nm, red:red, R:R) or different (blue:red, B:R or red:blue, R:B) wavelengths, or of white:white (W:W, controls). Photoperiodic induction occurred under R:R and B:R, but not under B:B and R:B light conditions; the W:W condition induced an intermediate response. The second set of SKPs used symmetrical light periods (L:D:L:D = 1:11:1:11 h), and measured effects also on the activity rhythm. Birds were first exposed to one of the four SKPs (R:R, B:B, R:B, or B:R) for three weeks, subsequently were released into dim constant light (LLdim; approximately 0.01 Wm(-2), the night light used in an L:D cycle) for two weeks, and then were returned to respective SKPs for another three weeks. Activity was greater in the R:R compared to B:B, and in B:R compared to R:B light condition. Zugunruhe (intense nighttime activity, indicating migratory restlessness in a caged situation) developed under the R:R and B:R, but not the B:B and R:B, light condition. Under LLdim, all birds free-ran with a period >24h, the Zugunruhe had a circadian period longer than the daytime activity, and the re-entrainment to SKPs was influenced by the position of light periods relative to circadian phase of the activity rhythm. Photoperiodic induction at the end of 8 weeks was found in the R:R and B:R, but not in B:B, light conditions; in the R:B condition only one bird had initiated testes. Taken together, these results suggest that in the blackheaded bunting, the circadian photoperiodic clock is differentially responsive to light wavelengths; this responsiveness is phase-dependent, and the development of Zugunruhe reflects a true circadian function. Wavelength-dependent response of the photoperiodic clock could be part of an adaptive strategy in evolution of the seasonality in reproduction and migration among photoperiodic species under wild conditions.     

Wavelength Dependency of Light-Induced Effects on Photoperiodic Clock in the Migratory Blackheaded Bunting (Emberiza melanocephala)

The effects of light wavelength on photoperiodic clock were determined in the migratory male blackheaded bunting (Emberiza melanocephala). We constructed an action spectrum for photoperiodic induction (body fattening, gain in body mass, and gonadal recrudescence) by exposing birds for 4.5 weeks to 13 h light per day (L:D = 13:11 h) of white (control), blue (450 nm), or red (640 nm) color at irradiances ranging from 0.028 to 1.4 W m^?2. The threshold light irradiance for photoinduction was about 10-fold higher for blue, compared to red and white light. Phase-dependent effects of light wavelength on the photoperiodic clock were further examined in the next two sets of skeleton photoperiods (SKPs). In the first set of SKPs, birds were exposed for four weeks to asymmetrical light periods (L:D:L:D = 6:6:1:11 h) at 0.25 ± 0.01 W m^?2; two light periods applied were of the same (450 nm: blue:blue, B:B; 640 nm, red:red, R:R) or different (blue:red, B:R or red:blue, R:B) wavelengths, or of white:white (W:W, controls). Photoperiodic induction occurred under R:R and B:R, but not under B:B and R:B light conditions; the W:W condition induced an intermediate response. The second set of SKPs used symmetrical light periods (L:D:L:D = 1:11:1:11 h), and measured effects also on the activity rhythm. Birds were first exposed to one of the four SKPs (R:R, B:B, R:B, or B:R) for three weeks, subsequently were released into dim constant light (LL_dim; ?0.01 W m^?2, the night light used in an L:D cycle) for two weeks, and then were returned to respective SKPs for another three weeks. Activity was greater in the R:R compared to B:B, and in B:R compared to R:B light condition. Zugunruhe (intense nighttime activity, indicating migratory restlessness in a caged situation) developed under the R:R and B:R, but not the B:B and R:B, light condition. Under LL_dim, all birds free-ran with a period >24 h, the Zugunruhe had a circadian period longer than the daytime activity, and the re-entrainment to SKPs was influenced by the position of light periods relative to circadian phase of the activity rhythm. Photoperiodic induction at the end of 8 weeks was found in the R:R and B:R, but not in B:B, light conditions; in the R:B condition only one bird had initiated testes. Taken together, these results suggest that in the blackheaded bunting, the circadian photoperiodic clock is differentially responsive to light wavelengths; this responsiveness is phase-dependent, and the development of Zugunruhe reflects a true circadian function. Wavelength-dependent response of the photoperiodic clock could be part of an adaptive strategy in evolution of the seasonality in reproduction and migration among photoperiodic species under wild conditions.     

Different circadian rhythms regulate photoperiodic flowering response and leaf movement in Pharbitis nil (L.) Choisy

The phase shifting effect of red light on both the leaf movement rhythm, and on the rhythm of responsiveness of photoperiodic flower induction towards short light breaks (10 min red light), has been studied in Pharbitis nil, strain Violet, and comparisons between the two rhythms have been made. The phase angle differences between the rhythms after a phase shift with 2 or 6 h of red light given at different times during a long dark period were not constant. The results indicate the involvement of two different clocks controlling leaf movement and photoperiodic flower induction.Abbreviations DD continuous darkness - l:D x:y light/dark cycles with x hours of light and y hours of darkness - PPR rhythm of photoperiodic responsiveness towards light break     

Effect of light pulses in early scotophase on resetting of the night-measuring diapause clock of the Indian meal moth, Plodia interpunctella

Abstract.  The Indian meal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae) may enter diapause in the last larval instar in response to the photoperiod during the preceding instars. An hourglass-type photoperiodic clock may measure night length for this purpose. The present study describes the resetting of the hourglass by light pulse(s) in the early scotophase and by scanning the subsequent clock phase by another light pulse (P). When the lights-off time of a first light pulse is fixed at 4 h after dusk under LD 4 : 20 h and LD 6 : 18 h photoperiods and its duration is increased from 1 to 3 h, the critical night length (CNL) from dawn is decreased, but that from dusk to P increases. A 3-h first light pulse efficiently resets the time measuring system. If this 3-h light pulse is split into two 1-h light pulses (L₁ and L₂) by 1 h of darkness, the dark-time measuring function appears to be impeded and CNL from P to dawn disappears, but that from L₂ to P is expressed. This indicates that the receptivity to light pulses varies among individual insects.     

Chloroplast Biogenesis: XXII. Contribution of Short Wavelength and Long Wavelength Protochlorophyll Species to the Greening of Higher Plants

The contribution of short and long wavelength membrane-bound fluorescing protochlorophyll species to the over-all process of chlorophyll formation was assessed during photoperiodic growth. Protochlorophyll forms were monitored spectrofluorometrically at 77 K during the first six light and dark cycles in homogenates of cucumber (Cucumis sativus L.) cotyledons grown under a 14-hour light/10-hour dark photoperiodic regime, and in cotyledons developing in complete darkness. In the etiolated tissue, short wavelength protochlorophyll having a broad emission maximum between 630 and 640 nm appeared within 24 hours after sowing. Subsequently, the long wavelength species fluorescing at 657 nm appeared, and accumulated rapidly. This resulted in the preponderance of the long wavelength species which characterizes the protochlorophyll profile of etiolated tissues. The forms of protochlorophyll present in etiolated cucumber cotyledons resembled those in etiolated bean leaves in their absorption, fluorescence, and phototransformability. A different pattern of protochlorophyll accumulation was observed during the dark cycles of photoperiodic greening. The short wavelength species appeared within 24 hours after sowing. Subsequently, the long wavelength form accumulated and disappeared. The long wavelength to short wavelength protochlorophyll emission intensity ratio reached a maximum (~3:1) during the second dark cycle, then declined during subsequent dark cycles. Short wavelength species were continuously present in the light and dark. Primary corn and bean leaves exhibited a similar pattern of protochlorophyll accumulation. In cucumber cotyledons, both the short and long wavelengths species appeared to be directly phototransformable at all stages of photoperiodic development. It thus appears that whereas the long wavelength protochlorophyll species is the major chlorophyll precursor during primary photoconversion in older etiolated tissues, both long wavelength and short wavelength species seem to contribute to chlorophyll formation during greening under natural photoperiodic conditions.     

Role de l'intensite lumineuse rouge (630–670 nm) dans le determinisme photoperiodique de la diapause nymphale de Pieris brassicae

In previous communications we have reported the role of red light on the determinism of the pupal diapause of Pieris brassicae (630 to 670 nm, wavelengths corresponding to the maximum of absorption of pterobilin, the blue bile pigment present in the integument and haemolymph).Red light suppresses the diapausing effect of the 9-hr/24-hr photophase of white light (identical energy in both and similar to the energy under natural conditions). The activity of red light in two experimental conditions: 9-hr white light followed by 15 hr of red lights of different intensities, and 9-hr red lights of different intensities followed by 15 hr darkness is examined. The photoperiodic determinism of pupal diapause is discussed.     

Effects of light wavelength on growth and survival rate in juvenile Pacific bluefin tuna, Thunnus orientalis

The spectral sensitivity of the fish and the suitable light wavelength range for survival and growth performance of juvenile Pacific bluefin tuna (PBT) were investigated. The spectral sensitivity peak of PBT under photopic condition was observed between 449 and 503 nm, which corresponded to their natural habitat. The fish were reared in tanks irradiated continuously with 4 kinds of light emitting diodes (LEDs). The maximum wavelength of LEDs used for the rearing experiment were 460 nm (blue), 520 nm (green), 630 nm (red), and 450–680 nm (white). There was no notable difference in survival rate among fish in the four LED groups. However, the growth of juvenile PBT was lesser under red light compared to the green and white light wavelengths. These results suggest that PBT juveniles have low sensitivity to red light because the fish are rarely exposed to the red light wavelengths under natural ocean conditions. Thus, low sensitivity to red light negatively influenced the feeding behavior and growth of PBT juveniles.     

Capacity of cytochrome and alternative path in coupled and uncoupled root respiration of Pisum and Plantago

A critical duration of darkness must be exceeded for the photoperiodic induction of flowering in short-day plants. This requires detection of the light/dark transition at dusk and the coupling of this information to a time-measuring system.
Lowering the P_fr/P_tot, ratio photochemically at the end of the day did not accelerate the onset of dark timing in Pharbitis nil Choisy cv. Violet. Time-measurement was initiated when, with no change in spectral quality, the irradiance fell below a threshold value. Thus, if the light/dark transition at dusk is sensed by a reduction in P_fr, this reduction can be achieved as rapidly through thermal reactions as through photochemical ones. When given at hourly intervals during a 6-h extension of a 24-h main light period in white light, pulses of red light were as effective as continuous red light in delaying the onset of timing; pulses every 2 or 3 h were less effective. The effectiveness of intermittent red light indicates that phytochrome is the photoreceptor and the requirement for frequent exposures suggests that P_fr is lost rapidly in the dark. However, the red light pulses could not be reversed by far-red light, which argues against this hypothesis. An alternative explanation is that the perception of light as being continuous occurs only when "new" P_fr is regenerated sufficiently frequently.
The nature of the coupling of the dusk signal to the time-measuring system is discussed and it is suggested that the effect of each red light pulse is to delay the phase of the photoperiodic rhythm by 1–3 h.     

Determination of effective light pulse and classical Bünsow experiment in the larval diapause of the Indian meal moth Plodia interpunctella

Plodia interpunctella Hübner (Lepidoptera: Pyralidae) comprises a model insect to analyse the photoperiodic time‐measuring system controlling its larval diapause. In the present study, the effective length of light pulse in night interruption experiments is determined at 25 °C. Various lengths of light pulse are tested by inserting them at the midnight of an LD 12 : 12 h photoperiod. When the light pulse is 15 or 30 min, the incidence of diapause is 86%. To inhibit the induction of diapause effectively, a light pulse of 1.75–2 h is needed. The incidence of diapause is 12% under an LD 12 : 5 : 2 : 5 h photoperiod. To determine the precise role of the light pulse, 2‐h light pulses placed at the midnight of an LD 12 : 12 h photoperiod are disrupted systematically by darkness. When a 2‐h light pulse is disrupted by 15 min of darkness, diapause is generally prevented (< 29%) regardless of the temporal position of darkness. Longer disruption by darkness induces diapause moderately (37–67%). A Bünsow experiment is also conducted at 25 and 20 °C, in which the main photophase of 12 h of light is combined with 24–72‐h scotophases scanned by a 2‐h light pulse. The photoperiodic cycle length tested, therefore, varies in the range 36–84 h. In each cycle length, the incidence of diapause fluctuates as the light pulse moves toward dawn. However, no regular and circadian changes in the percentage diapause are observed in relation to diapause determination.     

暗期干扰对棉铃虫两个不同地理种群滞育抑制作用的比较

至今,所测试昆虫的光周期反应均表明,光周期反应对暗期干扰高度敏感,短暂的光脉冲都可在不同程度上逆转长夜效应,抑制滞育的发生。在研究了棉铃虫Helicoverpa armigera泰安种群(36.15°N,116.59°E)和喀佐种群(41.34°N,120.27°E)光周期反应的基础上,在滞育诱导的短光周期下(L12:D12和L9:D15),分别测试了暗期不同时段1h光脉冲对这两个不同地理种群滞育抑制的影响。25和22℃下的光周期反应显示了泰安种群在长暗期11—14 h的滞育率均显著低于喀佐种群;泰安种群的临界暗长分别为11.7 h和11.5 h,喀佐种群分别为10.5 h和10.3 h,泰安种群均比喀佐种群长1.2 h。在所测试的暗期干扰实验中,除了极少数光脉冲干扰点外,泰安种群蛹滞育率显著低于喀佐种群,但两者的滞育反应曲线基本相似。在短光周期L9:D15下,泰安种群和喀佐种群均显示了光脉冲落入暗期的第9—11小时最有效地抑制了滞育的发生。在短光周期L12∶D12下,泰安种群和喀佐种群在25℃时均显示了光脉冲落入暗期的第3—4小时和第10小时导致了最低的滞育发生;但在22℃时,喀佐种群只在暗期的第3—4小时显示了最高的滞育抑制。这些结果揭示了偏南的泰安种群对暗期干扰的敏感性强于偏北的喀佐种群,但这两个地理种群的最高光敏感位点基本相同。     

Photoperiodic and light spectral conditions which inhibit circulating concentrations of thyroxine in the male hamster

Adult male Syrian hamsters were exposed daily for 12 weeks to 11 h/day of cool white fluorescent light (350 +/- 50 microW/cm2) followed by an additional 3 h of near ultraviolet (339-317 nm), blue (435-500 nm), green (515-550 nm), yellow (558-636 nm) or red (653-668 nm) light at an irradiance of 0.2 microW/cm2 or to total darkness. Animals exposed to the wavelengths between 558-668 nm (yellow or red half peak bandwidths) or those receiving a total of 13 h of darkness/day had suppressed circulating levels of thyroxine (T4), a depressed free T4 index (FT4I) and a higher T3/T4 ratio compared to animals receiving a total of 14 h of white light (350 +/- 50 microW/cm2). These results suggest that specific wavelengths of light can affect the neuroendocrine-thyroid axis.     

Photoperiod induced developmental effects on silkmoth,Bombyx mori

Bombyx mori is most common silkmoth used for the commercial production of the silk. The different environmental conditions may affect development of silkmoth and hence the output of silkworm. Present study investigated effects of different photoperiodic conditions on the development of silkmoth. Three experiments were performed on newly hatched larvae. In experiment1, larvae were divided into three groups; group 1, 8L:16D (8 h light and 16 h dark), group 2, 12L:12D (12 h light and 12 h dark) and group 3, 16L:8D (16 h light and 8 h dark). In experiment 2, larvae were exposed to either continuous light or continuous dark. In experiment 3, larvae were divided into three groups. Each group received light of similar duration (16L:8D) and intensity (1.1 ± 0.1 PAR) but of different wavelengths; i.e. long wavelength red light (640 nm), short wavelength blue light (450 nm) and white light. Larvae were fed on young mulberry (Morus alba) leaves. Temperature and humidity were maintained at 26 (±2) °C and 75 (±5)%, respectively, throughout the study. Observations on change in body length of larvae were made on every alternate day. We also calculated the shell ratio, time taken in cocoon formation and emergence. Results from experiment show that the larvae underwent significant change in body length but with faster growth rate under 12L:12D photoperiod in comparison to 8L:16D and 16L:8D. Time taken in cocoon formation and hatching was significantly less, while shell ratio was significantly higher in 12L:12D in comparison to other groups. We did not observe much effect of monochromatic light on the development of silkmoth.     

Spectral preference and temporal modulation of photic orientation by Colorado potato beetle on a servosphere

The Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), is a diurnal insect that strongly relies on visual cues to guide its walk. In the present study, we investigated the orientation behavior of non‐diapausing walking CPB in response to emissive colors produced by light emitting diodes (LEDs) in a dual choice arena adapted to a servosphere, where the only illumination available came from the photo‐stimuli. Our results demonstrate that CPB show positive phototactic behavior when stimulated with different wavelengths of light; they preferred to orient towards white (420–775 nm), ultraviolet (UV) (351 nm), blue (472 nm), green (570 nm), yellow (585 nm), orange (590 nm), and red (660 nm) over darkness when both alternatives were offered, but no orientation responses were elicited by infrared (940 nm). Both males and females preferred yellow and green over other colors, but did not show any particular preference between them, thus correlating with their preference for wavelengths reflected by vegetation. Sexual differences were noted in that male CPB preferred white over either red or UV, whereas female CPB did not show any preferences when offered these colors. Female CPB preferred UV and blue over red, whereas males showed no preferences when these colors were offered. Colorado potato beetles turned at higher angles and performed more tortuous walks in complete darkness and when infrared vs. darkness were offered compared with the rest of the colored lights. Both sexes preferred continuous over pulsed yellow light. Colorado potato beetles subjected to pulsed yellow light showed a temporal alteration of their walking performance by walking less, slower, and turning at a higher rate. The results are discussed with regard to the role of color in the CPB attraction to host plants and conspecifics as well as the role of intermittent photic stimuli in their orientation behavior. The information provided here provides a basis for the improvement of trapping devices for detection and survey of incipient or invasive CPB, and development of alternate control strategies for this important pest of potatoes and other solanaceous crops.     

The influence of light wavelength on reproductive photorefractoriness in migratory blackheaded bunting (Emberiza melanocephala).

There are two effects of long day length on reproductive responses in birds, one is the photoinduction of gonadal growth and maturation and the other is the induction of gonadal regression and photorefractoriness. Although it is likely that the same photoreceptors are involved in the photoinduction of gonadal growth and the onset and maintenance of photorefractoriness. and so the influence of wavelength should be similar, this has not been investigated. Therefore, we investigated the influence of light wavelength on reproductive photorefractoriness in the migratory male blackheaded bunting held under long photoperiods. In mid May, when photoperiod was approximately 14L:10D (14 hours light:10 hours darkness), eight groups of sexually mature birds were moved indoors on an artificial photoperiod of 14L:10D (L - 450 lux. D - 0 lux). Then after 3 weeks, for six groups, a 4-h light period in the morning (zt 0-4; zt 0 [zeitgeber time 0] refers to the beginning of lights-on period) or in the evening (zt 10-14) was substituted with green (428 nm), red (654 nm) or white light at 16 +/- 2 lux intensity. Of the remaining two groups, one was maintained on 14L: 10D and the other transferred to 10L:14D: these served as controls. At the end of 4 weeks, all birds were found to have undergone testicular regression, irrespective of LD cycle they were exposed to. When these gonadally regressed birds were subjected to 16L:8D for another 4 weeks, to test their responsiveness to the stimulatory effects of long day lengths, only those exposed to 10L:14D and 14L:10D with a 4-h green light period showed testicular regrowth. On the other hand, birds exposed to 14L:10D with a 4-h white or red light period remained fully regressed, similar to 14L:10D controls. Except for some individual difference, there was no difference in response between the groups that received a 4-h light period in the morning and that received it in the evening. These results suggest that the wavelengths of light influence induction of buntings from the photosensitive state into the photorefractory state. Whereas the short light wavelengths facilitated recovery from the photorefractoriness, the long light wavelengths were more effective in maintaining the photorefractoriness.     

Two phytochrome-dependent processes in Anagallis arvensis L.: flowering and stem elongation

Abstract Two phytochrome-dependent processes are compared in Anagallis arvensis, an absolute long-day plant: flowering and internode elongation. The effectiveness of red or far red radiation is different according to the length of the treatment and its position within the photoperiodic cycle: for both flowering and internode elongation, 15 h treatments with wavelengths above 700 nm are promotive, whilst wavelengths below 700 nm are inhibitory. In contrast, night breaks of 3 h red light (λ < 700 nm), given after the middle of the dark period, are promotive for flowering and inhibitory for internode elongation. These results are discussed in context with data reported in the literature on photoperiodic control of long-day plants.     

Different actions of red and blue or far-red lights in the photoperiodic tuberization of Begonia evansiana

The spectral dependence of Begonia evansiana in supplementarylight periods of photoperiodic tuberization and sprouting wasinvestigated. Supplementary application of red light inhibitedtuber development, thereby stimulating vegetative growth. Supplementaryblue or far-red light also suppressed tuber development, butbarely stimulated vegetative growth. However, both red and blue light, given at 6°C during themain light period or the supplementary light period, permittedthe tuberization under the subsequently given conditions ofeither long-days or darkness at 23°C. Blue light appliedafter 5-days of irradiation with white light at 10°C, showedalmost the same action as far-red light, which suppressed tuberizationin darkness. The nature and function of the pigments concernedin the photoperiodic responses are discussed. (Received October 11, 1968; )     

Oak Seedlings Grown in Different Light Qualities

Seedlings of oak (Quercus robur) were germinated in darkness for 3 weeks and then given continuous light or short pulses of light (5–8 min every day). The morphological development was followed during 25 days. In continuous white, blue, and red light the stem growth terminated after about 10 days by formation of a resting bud. At that time the seedlings were about 100 mm high. In con tinuous long wavelength farred light (wavelength longer than 700 nm) the stem growth including leaf formation was continuous without the formation of resting buds, and the stem length was about 270 mm after 25 days. The number of nodes developed became twice that of the seedlings grown in while light. The leaves became well developed in all light colours, but leaf areas were largest in plants cultivated in white light. Compared to dark grown seedlings the mean area per leaf was increased about five times in continuous long wavelength far red light. A supplement with short (5 min) pulses of red light each day increased the leaf area up to 20 times. The stem elongation showed a high energy reaction response, i.e. the stem length increased only in continuous long wavelength far-red light but was not influenced by short pulses of red light or far-red light. The leaf expansion, however, was increased by short pulses of red light with a partial reversion of the effect by a subsequent pulse of far-red light. The fraction of the plant covered with periderm was higher in plants given continuous light. In respect to periderm inhibition continuous long wavelength far red light was the most effective. The transfer of seedlings from darkness to continuous white light gave anthocyanin formation in the stem 10–20 mm below the apex. This formation took place in the cortex and was evident in plants grown in darkness or under short pulses of light. Plants grown in continuous red, blue or long wavelength Far red light showed only traces of anthocyanin.     

Circadian response reduction in light and response restoration in darkness: a "skeleton" light pulse PRC study in mice (Mus musculus)

Entrainment may involve responses to dawn, to dusk, and to the light in between these transitions. Previous studies showed that the circadian system responds to only 2 light pulses, one at the beginning and one at the end of the day, in a similar way as to a full photoperiod, as long as the photoperiod is less than approximately 1/2 tau. The authors used a double 1-h light pulse protocol with different intervals of darkness in between (1, 2, 4, 7, 10, and 16 h) to study the phase responses of mice. The phase response curves obtained were compared to full light pulse PRCs of corresponding durations. Up to 6 hours, phase responses induced by double light pulses are virtually the same as by a corresponding full light pulse. The authors made a simple phase-only model to estimate the response reduction due to light exposure and response restoration due to dark exposure of the system. In this model, they assumed a 100% contribution of the first 1-h light pulse and fitted the reduction factor for the second light pulse to yield the best fit to the observations. The results suggest that after 1 h of light followed by less than 4 h of darkness, there is a considerable reduction in response to the second light pulse. Full response restoration requires more than 10 h of darkness. To investigate the influence of the duration of light on the response saturation, the authors performed a second series of experiments where the duration of the 2 light pulses was varied from 4 to 60 min each with a fixed duration of the stimulus (4 h). The response to 2 light pulses saturates when they are between 30 and 60 min long. In conclusion, double pulses replace single full light pulses of a corresponding duration of up to 6 h due to a response reduction during light, combined with response restoration during darkness. By the combined response reduction and response restoration, mice can maintain stable entrainment to the external LD cycle without being continuously exposed to it.