Physiological Function of Night Interruption in Lemna paucicostata 6746: Action of Light as a Phaser on the Photoperiodic Clock

Flowering of Lemna paucicostata 6746 under short-day conditionsis completely inhibited by daily night interruption given tothe "inhibition zone" that starts at Zeitgeber time (ZT) 14,i.e., 14 h after dawn, and ends 14 h before the next dusk [Oota(1983a) Plant & Cell Physiol. 24: 327]. With a modifiedmin-SD method, most of these night interruptions were foundto signal the false dawn (false ZT 0) after one entraining cycle.Thus, on and after day 2 the interruption was associated withthe next main photoperiod to form a noninductive skeleton photoperiod.However, a light pulse applied at the start of the inhibitionzone, caused no phase shift in the photoperiodic clock, andformed a noninductive skeleton photoperiod in association withthe preceding main photoperiod. The complete floral inhibition due to the night interruptionwas ascribed to the illumination of both the LI-phase (realor false ZT 0) and L2-phase (real or false ZT 14), or the twolight-sensitive fractions of the original or shifted criticalphotoperiod, by the thus formed skeleton photoperiod, just aswas the case for the floral inhibition by complete photoperiodslonger than the critical daylength, 14 h [Oota (1983a), Oota(1983b) Plant & Cell Physiol. 24: 1503]. (Received October 20, 1983; Accepted January 7, 1984)     

Floral Inhibition in Lemna paucicostata 6746 Due to Night Interruption

The floral response to various 24-h photoperiodic cycles ofthe short-day plant, Lemna paucicostata 6746 was investigated.No day that had a main photoperiod longer than about 14 h wasable to induce flowers, evidence that the critical day lengthwas ca.14 h. Flowering in the 12-, 9- or 6-h day was inhibitedcompletely by a light pulse inserted daily in the ‘inhibitionzone’ that ranged from about 14 h after the precedingdawn to about 14 h before the next dusk. In the 3- and 1-h days,only the pulse applied 14 h after the dawn completely inhibitedflowering. These results suggest that the daily night interruption prohibitedflowering only when it was linked to either the preceding orthe subsequent main photoperiod to form a skeleton photoperiodwhose length was equal to, or longer than, the critical daylength. Analysis of the floral response to skeleton schedules11:13 and 13:11 on Pittendrigh's model of the photoperiodicclock indicated that light-on circadian oscillation probablyis involved in the day length measurement. ¹ Dedicated to the memory of Dr. Joji Ashida. (Received July 13, 1982; Accepted January 17, 1983)     

The action of skeleton photoperiods on flowering in Lemna perpusilla

The effects of 24 hr cycle skeleton photoperiodic schedulesinvolving two short light pulses on flowering in Lemna perpusillahave been studied. Simulation of complete photoperiods by correspondingskeletal ones is nearly perfect for all photoperiods up to 8hr and is unstable for periods of 9 to 13 hr. A jump in theresponse phase appears when skeleton photoperiods ranging from12 to 13hr are given. For all skeleton photoperiods longer than14 hr the phase is entrained so that it agrees with that givenby skeleton photoperiods of complemental lengths. That is, askeleton photoperiod of 18 hr is equivalent to that of 6 (=24–18) hr. Simulation is largely related to whether thesecond pulse is locked on to "dawn" or to "sunset" dependingon when it falls during the dark period following the firstpulse. The inductive action of skeleton photoperiods that gives unstableentrainment depends on the length of a preliminary dark periodgiven before the plant receives the first pulse, since in theseskeleton schedules the sensitive zone to the second pulse shiftswith the length of the preliminary darkness. Thus, we tentatively conclude that the circadian oscillationin L. perpusilla involves an entrainment mechanism and thatphotoperiodic induction is contingent on the coincidence oflight and a specific inductive phase in oscillation. (Received September 18, 1968; )     

Maintenance of high Pfr-level in the dark period in relation to flowering in Lemna gibba G3

The relative Pfr-level in a long-day duckweed, Lemna gibba G3,was estimated by the null response method. The null % R value(% R in a R/FR-mixture that provides a null flowering response.This value was assumed to indicate the endogenous Pfr-levelof the duckweed.) remained high during the initial hours ofthe 15 hr nyctoperiod then decreased rapidly, if a 12 or 33hr photoperiod preceded the nyctoperiod. The null % R valuedropped immediately after the start of the 15 hr nyctoperiodsubsequent to a 1 or 24 hr photoperiod. Thus, the duration ofthe maintenance of a high Pfr-level changed rhythmically dependingon the length of the preceding photoperiod. Nyctoperiods ofup to 9 hr following a 12 hr photoperiod hardly affected flowering,but nyctoperiods given after a 24 hr photoperiod suppressedflowering in proportion to the length of the period. The Pfr-levelin the nyctoperiod, therefore, seems to be important for flowering,and phytochrome change, as a function of the length of photoperiod,may serve as a photoperiodic timer. Although floral responseto interruption with R or FR changed with the application period,the difference in response between R-treated and FR-treatedplants was relatively constant during a 15 hr nyctoperiod combinedwith a photoperiod of any length other than 1 hr. Apparently,the floral response to the R or FR pulse was regulated by ashift in the Pfr-level caused by the light pulse. (Received April 2, 1979; )     

Rapid photoperiodic responses in Japanese quail: is daylength measurement based upon a circadian system?

Experimental photoperiods, presented either once only or repeatedly, were used to assess the oscillatory and hourglass properties of the photoperiodic clock in Japanese quail. Gonadectomized quail on 8-hr daylengths respond to a single skeleton photoperiod consisting of two 8-hr light pulses separated by 2 hr of darkness (i.e., LDLD 8:2:8:6) with a marked increase in secretion rate of luteinizing hormone (LH). This response suggests that the second light pulse interacts with a "photoinducible phase" (phi i) lying some 10-16 hr from "dawn" (start of the first light pulse). If, however, groups of quail maintained on 8-hr daylengths are transferred to continuous darkness (DD), and the position of the phi i is sought by a single 8-hr light pulse applied at various times on the first or third day of DD, then an increase in circulating LH is, at best, barely detectable. It would appear that a strongly responsive phi i does not recur rhythmically in DD. Instead, the light pulse apparently acts primarily as a "dawn" signal that triggers a single cycle of photoinducibility, since a second 8-hr light pulse, placed to begin 2 hr after the end of the first, induces a large increase in plasma LH. Similar results are obtained if any single 8-hr light pulse presented to animals held in darkness is preceded, 10 hr earlier, by a short "dawn" light signal. Such dawn signals can be effective when very short; a pulse of only 30 sec can cause a subsequent phi i. The dawn pulse is effective at any circadian phase and leads to a single cycle in photoinducibility. In contrast, a much longer light pulse (perhaps not less than 4 hr) is needed to interact with phi i if significant gonadotropin secretion is to be stimulated. In confirmation of the findings described above, we found that Nanda-Hammer lighting schedules have remarkably little effect in stimulating gonadotropin secretion in gonadectomized quail. There is, for example, a very marked difference between the effectiveness of "resonating" schedules such as LD 6:6, which stimulates a high LH secretion rate since each "inductive" light pulse is preceded by an appropriate "dawn" signal, and a theoretically effective schedule such as LD 6:30, which induces a very small response by comparison. Such schedules (even theoretically noninductive ones) can, however, be made very highly inductive if alternate light pulses are preceded by an appropriately positioned 15-min light pulse to act as "dawn."     

Photoperiodic requirements for flowering of the long-day duckweed, Lemna gibba G3

The min-LD estimation by the log. flower number vs. the cultureperiod curve provides a unique method of judging whether a givenphotoperiodic schedule is a long day or not for Lemna gibbaG3. Duckweeds in M-sucrose medium are exposed to the test scheduleat 26°C on either the first or second day of a continuouslight culture. If the min-LD (48 hr for control cultures) isnot changed, the inserted schedule is considered to have functionedas a long day. If, however, the min-LD is extended by 24 hr,the inserted schedule is judged to have functioned as a shortday. Examinations using this method of orderly designed light-darkschedules disclosed two critical phases in the light requirement;the initial and terminal 1 hr portions (designated the L1- andL2-phases) of the subjective day. Thus, a given day became along or short day when both the L1- and L2-phases were illuminatedor when either or both of the two phases were darkened. Thecritical daylength (11.5 hr) was just long enough to cover boththe L1- and L2-phases and the inductive phase (L2-phase) waslocated at the end of the subjective day. (Received June 9, 1975; )     

The role of the main photophase on dark-time measurement used for diapause determination in the Indian meal moth, Plodia interpunctella

Abstract.  The Indian meal moth Plodia interpunctella Hubner (Lepidoptera: Pyralidae) measures night length and enters diapause as a last-instar larva. To examine the role of photophase on dark-time measurement, the main LD 7 : 17 h photoperiod is disrupted by various lengths of darkness at 25 °C. When the light phase is not disrupted, the incidence of diapause is 76%. As the dark pulse disrupting a 7-h photophase becomes longer, the incidence of diapause decreases. To detect the dynamic kinetics of the time-measuring process, the main scotophase of 17 h is scanned by a 2-h light pulse. When the dark pulse in a 7-h photophase is fixed at 1 h after dawn and its duration is varied systematically from 1 to 3 h, or when the end of the dark pulse is fixed at 1 h before dusk, diapause is prevented completely by a 2-h light pulse inserted in the middle of 17-h darkness. These results are compared with those of a single night interruption of a 17-h scotophase with a 2-h light pulse but with an intact 7-h photophase. The disruption of a 7-h photophase by a dark pulse shifts the descending and ascending slopes of the response curve to some extent toward dawn and dusk, respectively, indicating that the dark pulse tends to shorten the critical length of dark time for diapause induction. When the main photophase (7 h) is interrupted by a 1-h dark pulse at 3–4 h after dawn, the 2-h scanning light pulse in the main scotophase (17 h) appears to act effectively as a dusk signal in the early scotophase. However, those in the mid- and late scotophase do not define the critical night length from dusk as sharply as for the critical night length from a 2-h light pulse to dawn. The results indicate the importance of photophase in the dark-time measurement.     

Effect of a light pulse during the dark on photoperiodic regulation of the rate of thyroxine-induced, spontaneous, and prolactin-inhibited metamorphosis in Rana pipiens tadpoles

Since Rana pipiens tadpoles injected with thyroxine (T4) early in the dark develop more slowly than those injected in the light, we studied the effect of giving a light pulse of 1 hr early in the dark. Tadpoles injected under a 7.5-W red light bulb in a darkened room with 0.2 microgram T4 daily at 2200 hr went through metamorphosis faster on a 12L:3D:1L:8D cycle with a light pulse after injection than on a 12L:12D cycle without a light pulse, and even faster on a 12L:1.5D:1L:9.5D cycle with a light pulse before the injection. Thus a 1-hr light pulse counteracted the metamorphic delay resulting from administration of T4 in the dark, and set in motion the conditions that resulted in a more rapid response to an injection of T4. However, a 1-hr light pulse in the early dark had no effect on growth and development of older or younger untreated tadpoles or those constantly immersed in 30 micrograms/liter T4. Larvae on 21L:3D with T4 injection in the dark and on 12L:3D:1L:8D with T4 injection at 0700 hr just before the start of the main light phase progressed faster than 12L:3D:1L:8D with injection at 2200 hr in the dark before only a 1-hr light pulse. Thus the length of the light phase immediately after T4 injection was significant. There was no difference on 12L:12D and 12L:3D:1L:8D cycles in the effectiveness of daily injections of 10 micrograms prolactin (PRL) in the early dark at 2200 hr in promoting tail growth or antagonizing tail resorption induced by T4 immersion. Under these conditions, PRL utilization did not appear to be inhibited by the light pulse.     

暗期干扰对环带锦斑蛾滞育诱导的影响

在短日照条件下,环带锦斑蛾Pseudopidorus fasciata以4龄幼虫进入冬季滞育,其滞育诱导的临界暗长为10.5 h。本文系统测试了暗期干扰实验［即在暗期采用单一的光（亦称光脉冲）干扰光周期反应的实验］对环带锦斑蛾幼虫滞育的抑制影响。幼虫孵化后在25℃,短日照L9∶D15(光9 h∶暗15 h),L10∶D14、L11∶D13及L12∶D12暗期的不同时段给予1 h或30 min光脉冲干扰。结果表明： 滞育是否被抑制主要取决于暗期的长度,当光脉冲干扰前的暗长（D₁）或光脉冲干扰后的暗长（D₂）超过临界暗长（10.5 h）时,100%的幼虫被诱导进入滞育;当D₁和D₂都短于（或等于）临界暗长时,滞育不同程度被抑制,抑制的效果则取决于所干扰的光周期、光脉冲的长度和光脉冲落入的位点。结果说明该虫滞育诱导的暗期干扰反应是基于临界暗长的时间测量。     

Physiological Structure of the Critical Photoperiod of Lemna paucicostata 6746

The critical day length or the length of the critical photoperiodfor the short-day duckweed, Lemna paucicostata 6746 is about14 h (Oota 1983). With the min-SD method, I found that not thewhole critical photoperiod but only its initial and terminalbrief fractions, called respectively the LI- and L2-phases,need be illuminated for a given day to be a noninductive day.Inversely, the darkened LI- and/or L2-phase makes the day inductive.The rest of the day can be either darkened or illuminated withoutmodifying the inductive or noninductive property of the day. Thus, the physiological structure of the critical photoperiodfor L. paucicostata 6746 closely resembles that of the criticalphotoperiod for the long-day duckweed, L. gibba G3 (Oota 1981). (Received May 24, 1983; Accepted September 21, 1983)     

The effect of temperature and light pulses on the induction of diapause in the Toyama strain of the Indian meal moth, Plodia interpunctella

Abstract.The photoperiodic response in Plodia interpunctella collected at Toyama (36.7°N) was of long-day type and highly sensitive to temperature. The critical photoperiod giving 50% diapause was between 14 and 16 h at 20°C, between 12 and 14 h at 25°C and between 6 and 8 h at 30°C. Effects of night interruption by a 2-h light pulse on the diapause response were examined at 25°C on different background photoperiods ranging from LD 12:12 h to LD 2:22 h. Percentage diapause was very low when the middle portion of dark period was interrupted, so that U- or V-shaped response curves were obtained with background scotophases longer than 12 h. In these curves, the descending slopes were less steep than the ascending slopes. The critical dark period measured from dusk to an interrupting light pulse was about 1.5 h longer than the critical dark period ( c . 10 h) in the normal photoperiodic response. The critical dark period from the interrupting light pulse to dawn, on the other hand, was not parallel to dawn but shorter than the normal critical period in LD 12:12 h and LD 10:14 h and longer than that in LD 7:17 h to LD 4:20 h, indicating that the priming effects of the light pulse might be under the influence of the photophase.     

Relationships between behavioral rhythms, plasma corticosterone and hypothalamic circadian rhythms

Circadian rhythms in physiological processes and behaviors were compared with hypothalamic circadian rhythms in norepinephrine (NE) metabolites, adrenergic transmitter receptors, cAMP, cGMP and suprachiasmatic nucleus (SCN) arginine vasopressin (AVP) in a single population of rats under D:D conditions. Eating, drinking and locomotor activity were high during the subjective night (the time when lights were out in L:D) and low during the subjective day (the time when lights were on in L:D). Plasma corticosterone concentration rose at subjective dusk and remained high until subjective dawn. Binding to hypothalamic alpha 1- and beta-adrenergic receptors also peaked during the subjective night. Cyclic cGMP concentration was elevated throughout the 24-hr period except for a trough at dusk, whereas DHPG concentration peaked at dawn. Arginine vasopressin levels in the suprachiasmatic nucleus peaked in the middle of the day. No rhythm was found either in binding to the alpha 2-adrenergic receptor, or in MHPG or cAMP concentration. Behavioral and corticosterone rhythms, therefore, are parallel to rhythms in hypothalamic alpha 1- and beta-receptor binding and NE-release. Cyclic GMP falls only at dusk, suggesting the possibility that cGMP inhibits activity much of the day and that at dusk the inhibition of nocturnal activity is removed. SCN AVP, on the other hand, peaking at 1400 hr, may play a role in the pacemaking function of the SCN that drives these other rhythms.     

Relationships Between Behavioral Rhythms,Plasma Corticosterone and Hypothalamic Circadian Rhythms

Circadian rhythms in physiological processes and behaviors were compared with hypothalamic circadian rhythms in norepinephrine (NE) metabolites, adrenergic transmitter receptors, cAMP, cGMP and suprachiasmatic nucleus (SCN) arginine vasopressin (AVP) in a single population of rats under D: D conditions. Eating, drinking and locomotor activity were high during the subjective night (the time when lights were out in L: D) and low during the subjective day (the time when lights were on in L: D). Plasma corticosterone concentration rose at subjective dusk and remained high until subjective dawn. Binding to hypothalamic α₁- and β-adrenergic receptors also peaked during the subjective night. Cyclic cGMP concentration was elevated throughout the 24-hr period except for a trough at dusk, whereas DHPG concentration peaked at dawn. Arginine vasopressin levels in the suprachiasmatic nucleus peaked in the middle of the day. No rhythm was found either in binding to the α₂-adrenergic receptor, or in MHPG or cAMP concentration. Behavioral and corticosterone rhythms, therefore, are parallel to rhythms in hypothalamic α₁-and β-receptor binding and NE-release. Cyclic GMP falls only at dusk, suggesting the possibility that cGMP inhibits activity much of the day and that at dusk the inhibition of nocturnal activity is removed. SCN AVP, on the other hand, peaking at 1400 hr, may play a role in the pacemaking function of the SCN that drives these other rhythms.     

Night length measurements by the circadian clock controlling diapause induction in the mosquito Aedes atropalpus

Night interruption experiments were used to investigate the behavior of the clock controlling diapause induction in the mosquito, Aedes atropalpus. The data from these experiments indicated that the clock included a circadian oscillator which was phase set at dusk. Following this event the oscillator proceeded to drive a nightly rhythm of sensitivity to light. This rhythm included a photoinducible phase where light interruption inhibited diapause. The photoinducible phase was fixed, occurring 7 to 9 hr after dusk in all photoperiod regimens tested. The photoinducible phase was followed by a refractory phase, which continued until dawn. During the refractory period light did not inhibit diapause. These observations indicated that the circadian clock behaved like an interval timer which was set at dusk. The rhythm of sensitivity to light, an inherited time scale, limited the induction of diapause to seasonal periods when nights were longer than 9 hr. As a result, diapause was induced only when the daylength dropped below the critical photoperiod of L15:D9 (hours of light:hours of dark).A ‘T’ experimental design was used to confirm the importance of the length of the night in clock controlled induction of diapause in this mosquito.     

Effect of photoperiod on incubation period in a wild passerine,Sylvia atricapilla

Time required for avian embryos to develop is influenced by incubation temperature and the amount of time adults incubate eggs. Experiments on poultry indicate that photoacceleration, the light‐induced stimulation of embryonic development, decreases the length of the incubation period as embryos receive more light. We hypothesized that eggs of wild birds exposed to longer periods of light should also have shorter incubation periods. We tested whether photoacceleration would occur in a species of open‐cup nesting passerine, the blackcap Sylvia atricapilla. We artificially incubated blackcap eggs under four different photoperiods, four hours of light (4L) and 20 h of dark (20D), 12L:12D, 20L:4D, and a skeleton photoperiod (1 h light, 2 times per day) that framed a 20 h day. While incubation periods were accelerated with increasing photoperiod length, the differences among photoperiods of 4, 12 and 20L were weak. Embryos exposed to skeleton photoperiods developed as fast as those exposed to 20L and significantly faster than those exposed to 4L and 12L treatments. Skeleton photoperiods may most closely approximate natural patterns of light exposure that embryos experience during dawn and dusk incubation recesses typically associated with adult foraging. If our results from this species also occur in other wild birds, exposure to different day lengths may help explain some of the variation in the observed seasonal and latitudinal trends in avian incubation period.     

Photoperiodic induction in quail as a function of the period of the light-dark cycle: implications for models of time measurement.

The earliest detectable event in the photoperiodic response of quail is a rise in luteinizing hormone (LH) secretion beginning at about hour 20 on the first long day. The timing of this rise was measured in castrated quail after entrainment to short daylengths which cause significant phase angle differences in the circadian system: (1) LD 2:22 and LD 10:14, and (2) LD 3:21 (T = 24 hr) and LD 3:24 (T = 27 hr). The quail were then exposed to 24 hr of light (by delaying lights-off), and the time of the first LH rise was measured; it was similar in all schedules. Quail were also entrained to LD 3:21 or LD 3:24 and then given a single 6-hr nightbreak 6-12, 7-13, or 13-19 hr after dawn. The earlier pulse was marginally more inductive in the 27-hr cycle. Thus the entrainment characteristics of the photoinducible rhythm (phi i) in quail appear very different from those of the locomotor circadian rhythm, and raise doubts as to whether phi i is a primary circadian oscillator.     

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.     

Aspects of the induction of diapause in a laboratory strain of the mite Tetranychus urticae

Some diapause characteristics were studied in a strain of the spider mite. Tetranychus urticae. which had been reared on bean plants in the laboratory for over 15 yr. The diapause induction response curve was of the long-day type, showing a sharply defined critical daylength of 13 hr 50 min. In constant darkness no diapause induction occurred, but with a photoperiod of 1L:23D diapause incidence was already complete. A thermoperiod with a 5°C amplitude induced diapause in combination with a short-day photoperiod only when the low phase of the thermoperiod coincided with the scotophase. The same thermoperiod did not induce any diapause in constant darkness. The photoperiodic reaction of the laboratory strain used in these experiments appeared to remain constant over a very long period of time and to be independent of the diapause history of previous generations of mites.Although photoperiodic sensitivity was demonstrated during the whole postembryonic development, sensitivity was maximal at the end of the protonymphal instar and declined rapidly during the deutonymphal instar. Only 2 inductive cycles of 10L:14D were required to induce up to 62% diapause if the mites were kept in continuous darkness during the remainder of their development. Long days or continuous light could reverse the inductive effect of a sequence of short-day cycles previously applied to the mites.Light breaks of 1 hr duration applied at different times during the dark period of a 10L:14D photoperiod generated a sharp bimodal response curve with two discrete points of sensitivity to the light breaks at 10 hr after ‘dusk’ and 10 hr before ‘dawn’, thus showing a remarkable similarity with the results obtained in light break experiments with some species of insects.     

Removal by chemicals of photoperiodic light requirements of Lemna gibba G3

Both the initial and the terminal 1 hr portions of the subjectiveday fraction, namely the L1- arid L2-phases, of a 24 hr daymust be illuminated in order for the day to be perceived asa long day in the min-LD determination by the long-day plant,Lemna gibba G3 (9). The light requirement of the L1-phase wassatisfied by a 10 min red light pulse given at the beginningof the phase. The red light effect was erased by a subsequent10 min far-red light, indicating phytochrome-mediated processesoccurring in the L1-phase. The light requirement of the L2-phasewas satisfied by blue or far-red light given during the terminal10 min period of the phase; there was no indication of phytochromeinvolvement. The light action on the L1-phase was replaced by10^–5 M of cyclic AMP or 10^–7 M of DL-isoproterenol.The isoproterenol action was antagonized by 10^–7 M ofDL-propranolol. Cyclic AMP (10^–5 M) combined with salicylicacid (10^–6 M), which can remove the light requirementof the L2-phase (10), rendered a completely dark day physiologicallyequivalent to a long day. Acetylcholine (10^–5 M) exertednyctomimetic action on the L1-phase of the second light day.The action of acetylcholine was antagonized by cyclic AMP (10^–5M). The L2-phase required no light in the presence of 10^–7M of DL-propranolol, and this propranolol action was not affectedby isoproterenol. These findings suggest changes in membranepermeability caused by the light given during the L1- and L2-phases. (Received July 7, 1976; )