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.     

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.     

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.     

Responsiveness to light of the circadian clock controlling eclosion in the blowfly, Lucilia cuprina

ABSTRACT. Eclosion in Lucilia cuprina (Wiedemann) occurs near dawn. The rhythm of eclosion persists in both darkness and constant light of high intensity (490μW cm^-2) with a period close to 24h. The sensitivity to light of the circadian clock controlling eclosion varies greatly according to the stage of the life cycle. During larval life the free running rhythm in darkness can be phase shifted by light pulses of 100μW cm^-2 intensity, with the transition from a Type 1 phase response curve to a Type 0, occurring with pulses of between 1 and 8h. Extending the last light period of LD to 24 h followed by constant darkness resets the phase of the rhythm by 12h, a transition from constant light to constant darkness initiates rhythmicity in flies made arrhythmic by being reared from eggs collected from adults maintained in constant light. After pupariation, the rhythm is relatively insensitive to light. Rhythmicity is sometimes induced by a transition from constant light to constant darkness, but the phase of the rhythm is not shifted by extending the last light period of LD before entering constant darkness. Repeated LD cycles applied after pupariation initiate and entrain the rhythm.     

Phase-shifting effects of a light pulse interrupting scotophase on locomotor activity rhythm and photoperiodic time measurement for diapause in the onion fly, Delia antiqua

When a light pulse of 1 h duration was given 3 h after lights off in a photoperiod of 11 h light : 13 h dark (LD 11 : 13) at 20°C, the phase of the major peak of locomotor activity rhythm in Delia antiqua was delayed for approximately 0.6 h. In contrast, it was advanced by approximately 0.6 h by a light pulse given 9 h after lights off. It is suggested that in the circadian clock, a pulse falling in the early scotophase is taken as a new dusk and a pulse falling in the late scotophase is taken as a new dawn. Although a sharply defined critical photoperiod did not exist in the diapause response to photoperiod in D. antiqua, the percentage of pupal diapause decreased by these pulses in LD 11 : 13 at 20°C. The effect of a 15 min light pulse on both locomotor activity rhythm and pupal diapause induction was stronger at 3 h than at 9 h after lights off, while a 1 min light pulse was ineffective at both times. The parallel effects of light pulse on locomotor activity rhythm and diapause response might be based on the same chronobiological functions.     

Effects of light pulses on the locomotor activity rhythm of Orchestia montagui (Amphipoda,Talitridae)

Animals of the amphipod Orchestia montagui are kept in constant darkness with two short light pulses. One pulse is applied at the beginning of subjective night (around the dusk) and the other one at the end of subjective night (around the dawn). The pulse duration is estimated in the order of one or two hours around the dusk as well as the dawn. The locomotor activity rhythm was monitored in individual animals in summer under constant temperature. Results revealed that whatever the experimental conditions, under continuous or interrupted darkness by pulses, two endogenous components have been highlighted. In fact, Periodogram analysis showed the presence of ultradian and circadian periods around 12 and 24 h, respectively. The shortest circadian period and the most important inter-individual variability was observed under pulse of 2 h around the dusk with mean value equal to τ_DD+pulse = 24h38′ ± 4h34′. The activity profiles are in majority unimodal. Moreover, the most activity peak showed a slipping of its location from the middle of subjective night under constant darkness to the middle of subjective day under pulse. Globally, the locomotor activity rhythm of O. montagui was better defined under pulses and specimens were significantly more active under continuous darkness. Moreover, a great variability around the activity time was observed especially with pulse of 1 h.     

Laboratory studies on the effects of pollen from Bt-maize on larvae of some butterfly species

Abstract:   Three lepidopteran species were tested to determine their susceptibility against the ingestion of pollen from genetically modified maize plants. To prove the existence of dose–response relations between the applied amount of pollen (Bt-maize) and the damages on the tested larvae, a method was developed which makes it possible to feed caterpillars with defined amounts of pollen. If their food plants were contaminated with pollen of a cultivar of the Bt-176 maize-line Pieris brassicae , Pieris rapae and Plutella xylostella -larvae fed less, grew more slowly and showed a higher mortality than caterpillars of an untreated control group. The 50% lethality (LD₅₀)-values were calculated for P.xylostella (L₄) with 19.2, for P. rapae (L₂) with 39.0 and also for P. brassicae (L₂) with 139.2 pollen of the transgenic maize Pactol CB. Studies with P. brassicae -caterpillars of different larval stages indicated, that older individuals showed a higher tolerance against pollen from Bt-maize than younger ones. It must be stated on the basis of the present studies, that ingestion of non-transgenic maize pollen has neither a positive nor a negative effect on caterpillars. It becomes clear from the information presented here that it is still difficult to make general statements about the endangering of butterflies, arising from cultivation of genetically modified maize lines. Further investigations on this issue are needed. Initially, the LD₅₀-values concerning the larvae of certain butterfly species have to be determined to anticipate the risks, and in addition the distances between habitats with caterpillar host plants and maize fields, and the abundance of these plants have to be considered.     

Night-interruption experiments and action spectra for dawn and dusk in relation to the photoperiodic clock of the cabbage whitefly,Aleyrodes proletella (Homoptera: Aleyrodidae)

The dark period (scotophase) is the most photoperiodically important part of a light-dark cycle in Aleyrodes proletella. Night-interruption studies have revealed three distinct dark stages: the photosensitive stage 1 lasts for about 3 h after dusk and 1-h light breaks both stop and re-set the photoperiodic clock; stage 2 also lasts about 3 h, but is photorefractory to some degree; stage 3 is photosensitive, but short light breaks do not re-set the clock although a 4-h light break (equivalent to a main photophase) does restore the capacity to respond to a normal critical night length in the post-interruption scotophase.Action spectra revealed peak photoperiodic sensitivity to blue light (410–430 nm) with 50% responses., at 1.5 μWcm⁻² and 2.5 μWcm⁻² for the dusk and dawn peaks respectively. These data are consistent with the view that the photopigment is a carotenoprotein.The results are interpreted in terms of the photoperiodic clock in A. proletella operating on the hour glass principle.     

Photokinetic responses and diel activity of sea lamprey Petromyzon marinus ammocoete larvae

Twenty-four ammocoete larvae of the sea lamprey Petromyzon marinus were tested individually for 3-day periods in electronic shuttleboxes (Ichthyotrons) with natural lighting through windows, to determine their diel activity pattern in terms of hourly percentages of the 24-h total. Mean hourly activity was slightly greater during the day (4–7 % h⁻¹) than at night (3·7% h⁻¹). Peak hourly activity (7·2% h⁻¹) occurred during the dawn crepuscular period as light levels were increasing, while the minimum hourly activity (1·6 % h^-l) occurred during the dusk crepuscular period while light levels were decreasing. The observed activity pattern is atypical of other aquatic animals similarly tested, and might be explained as an extraptically mediated photoinetic response.     

Evaluation of Serangium n. sp. (Col., Coccinellidae), a predator of Bemisia tabaci (Hom., Aleyrodidae) on cassava

Abstract  The potential of a new, previously unidentified Serangium species (Col., Coccinellidae) to control the high Bemisia tabaci (Gennadius) (Hom., Aleyrodidae) populations on cassava was evaluated. Field and laboratory studies were carried out to determine the abundance and feeding capacity of this Serangium species feeding on B. tabaci on cassava. Serangium nymphs and adults were most abundant in cassava fields late in the season, rising sharply from 5 months after planting (MAP) to a peak at 7–8 MAP. Pre-imaginal development averaged 21.2 days and was longest in eggs and shortest in the L₁ instar. Mean total prey consumption of immature Serangium increased with the stage of development with the lowest consumption in the L₁ instar and highest in the L₄ instar. Mean daily consumption was lowest on the first day after hatching in the L₁ instar and rose to a peak on the 13th day after hatching in the L₄ instar. Each Serangium larva consumed a mean of over 1000 nymphs during its entire development. These results have demonstrated the potential of this Serangium species to control B. tabaci populations on cassava.     

The effects of initial length and body curvature on shrinkage of juvenile Atlantic salmon during freezing

Fork length was measured in two groups of salmon parr (32–139 mm, frozen in a straight posture and frozen in a curved posture) before (L₁) and after (L₂) freezing and thawing. All the fish shrank. The decrease in length was significantly greater in the curved fish than the straight fish. The absolute reduction in length (L₁–L₂) was related directly to L₁, whereas the percentage reduction in length [(L₁–L₂)/L₁× 100] was related inversely to L₁.     

Duration times of the immature stages of Cacopsylla pyri L. (Hom., Psyllidae), estimated under field conditions, and their relationship to ambient temperature

The duration of the immature stages of Cacopsylla pyri L. was studied under field conditions by artificially infesting pear branches on several dates during the year. The duration of the egg stage decreased from winter to summer, as the season progresses and temperature rises, and slightly increased in September. It ranged from 27.4 to 6.7 days. The same trend was observed in the duration of the first three larval stages (L₁₋₃) which ranged from 18.8 to 10.3 days. For eggs deposited during the period February–August the duration of the last two larval stages (L₄₋₅) ranged from 17.5 to 12.1 days. However, the duration of L₄₋₅ developed from eggs deposited in September and which give rise to winter-form adults were the longest observed. The rate of egg development was related to average ambient temperature with a highly significant linear relationship. This relationship indicates that the egg stage requires a constant number of 158.9 (SD = 5.0) of day-degrees above an average temperature of 2.31°C to complete development. The rate of development both of L₁₋₃ and L₄₋₅ were related to average ambient temperature with curvilinear relationships. These relationships indicate a proportional increase in the developmental rate as temperature rises between 10–22°C. At the higher average temperatures that occurred in the summer experiments (24–27°C) the acceleration of development of L₁₋₃ is reduced and the developmental rate of L₄₋₅ decreases. The developmental rates of L₄₋₅ developed from eggs deposited in September did not follow the established relationship with temperature and they were lower than those in the other periods of the year with the same average temperature.     

Properties of two pectin lyases produced by Aureobasidium pullulans LV 10

Two pectic lyases, L₁ and L₂, from culture liquids of Aureobasidium pullulans LV 10 were partially purified by ultrafiltration, CM-Sepharose 6B, DEAE-cellulose and/or Sephadex G 100 column chromatography, and characterized. L₁ and L₂ showed optimum activity at pH 5 and 7.5 respectively, and at 40°C. The molecular weights of the enzymes determined by gel filtration were estimated to be 89000 1000 and 55000 1000 for L₁ and L₂ respectively. Both lyases were activated by Ca²⁺ ions. L₁ attacked highly esterified pectins, L₂ attacked low methoxy-pectins in preference to polygalacturonic acid.     

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.     

THE HAMSTER CLOCK PHASE-RESPONSE CURVE FROM SUMMERLIKE LIGHT:DARK CYCLES AND ITS ROLE IN DAILY AND SEASONAL TIMEKEEPING

We address the subject of entrainment of the hamster clock by the day:night cycle in summer when the sun sets after 6 PM and rises before 6 AM (nights<12 h). Summer day:night cycles were simulated by 6 light:dark (LD) cycles with D<12 h (summerlike, SLD) ranging from SLD 12.5h:11.5h (D, 6:15 PM–5:45 AM) to 18h:6h (D, 9 PM–3 AM). These are the near limiting SLDs for constant PM timing (entrainment) of behavioral estrus and wheel running in hamsters. The onset of estrus was observed every 4 d in the same hamsters as a phase marker of their 24h clock. On the day before an experimental estrus, preceded and followed by control onsets, a dark period was imposed to cover a putative 6 PM–6 AM light-sensitive period (LSP). This was scanned with a light pulse (and periodic 5sec bell alarms) lasting 5–240 min. Shifts in onset of estrus on the next day were plotted vs. the end of the light pulse for PM times (“dusk”) and its onset for AM times (“dawn”). The resulting phase shifts from the six SLDs were similar, permitting their combination into a single phase-response curve (PRC) of 1605 shifts. This SLD composite PRC rose at 10:15 PM, peaked at 2 AM (81min advanced shift), fell linearly to 5:55 AM, and then abruptly to normal at 6 AM (no shift). Peak shift was unaffected by light pulse duration or intensity, or hamster age. The SLD composite PRC lacked the 6 PM–9 PM curve of delayed shifts present in reported PRCs from LD 12h:12h and DD. However, a two-pulse experiment showed that all light from 6 PM to L-off was needed to block (balance) the advancing action of a 5min morning light pulse, thereby maintaining entrainment. A working hypothesis to explain daily entrainment and seasonal fertility in the golden hamster is illustrated. A nomenclature for labeling the phases of the hamster clock (circadian time) is proposed.     

Triiodothyronine Is a High-Affinity Inhibitor of Amino Acid Transport System L1 in Cultured Astrocytes

Abstract: The relationship between the transport of thyroid hormones and that of amino acids was examined by measuring the uptake of amino acids that are characteristic substrates of systems L, A, and N, and the effect of 3,3',5-triiodo-L-thyronine (T₃) on this uptake, in cultured astrocytes. Tryptophan and leucine uptakes were rapid, Na⁺-independent, and efficiently inhibited by T₃ (half-inhibition at ∼ 2 μ M ). Two Na⁺-independent L-like systems (L₁ and L₂), common to leucine and aromatic amino acids, were characterized kinetically. System L₂ had a low affinity for leucine and tryptophan ( K _m= 0.3–0.9 m M ). The high-affinity system L₁ ( K _m∼ 10 μ M for both amino acids) was competitively inhibited by T₃ with a K _i of 2–3 μ M (close to the T₃ transport K _m). Several T₃ analogues inhibited system L₁ and the T₃ transport system similarly. Glutamine uptake and α-(methylamino)isobutyric acid uptake were, respectively, two and 200 times lower than tryptophan and leucine uptakes. T₃ had little effect on the uptakes of glutamine and α-(methylamino)isobutyric acid. The results indicate that the T₃ transport system and system L₁ are related.     

Photoperiodic clock of diapause induction in Pseudopidorus fasciata (Lepidoptera: Zygaenidae)

Pseudopidorus fasciata enters diapause as fourth instar larvae at short day lengths. Using 24-h light-dark cycles, the photoperiodic response curves in this species appeared to be similar with a critical night length of 10.5h at temperatures below 30 degrees C. At an average temperature of 30.5 degrees C, the critical night length had shifted to between 15 and 17h. In experiments using non-24-h light-dark cycles, it was clearly demonstrated that the dark period (scotophase) was the decisive phase for a diapause determination. In night interruption experiments using 24-h light-dark cycles, a 1-h light pulse at LD12:12 completely reversed the long night effect and averted diapause in all treatments. At LD 9:15 light pulses of 1-h, 30- or 15-min also averted diapause effectively when both the pre-interruption (D(1)) or the post-interruption scotophases (D(2)) did not exceed the critical night length. If D(1) or D(2) exceeded the critical night length diapause was induced. The most crucial event for the photoperiodic time measurement in this species is the length of the scotophase. A 10-min light pulse placed in the most photosensitive phase reversed diapause in over 50% of the individuals. Night interruption experiments under non-24-h light-dark cycles indicated that the photoperiodic clock measured only D(1) regardless of the length of D(2), suggesting that the most inductive cycles are often those in which L+D are close to 24h. In resonance experiments, this species showed a circadian periodicity at temperatures of 24.5 or 26 degrees C, but not at 30.5 and 23.3 degrees C. On the other hand, Bünsow and skeleton photoperiod experiments failed to reveal the involvement of a circadian system in this photoperiodic clock. These results suggest the photoperiodic clock in this species is a long-night measuring hourglass and the circadian effect found in the final expression of the photoperiodic response in the resonance experiments may be caused by a disturbing effect of the circadian system in unnatural regimes.     

Genetic manipulation of the circadian clock's timing of sexual behaviour in the Queensland fruit flies, Dacus tryoni and Dacus neohumeralis

ABSTRACT. Mating in Dacus tryoni is restricted to dusk, whereas that of a sibling species, Dacus neohumeralis , occurs in the middle of the day. The timing of sexual behaviour in both species is determined by an interaction between a circadian clock and light intensity. In D. tryoni peak mating responsiveness is at the time of dusk, and the optimal light intensity for mating is approximately 91x. In D.neohumeralis peak responsiveness is in the middle of the day, and the optimal light intensity for mating is greater than 10 000 lx. The two species were crossed and the time of mating and response to light intensity of F₁, F₂ and backcross progeny determined. The circadian clock set a mating phase ('gate') as narrow in F₁ flies as in their parents, suggesting the circadian timing mechanism to be common between the two species. The results indicate that the genetic mechanism controlling timing is independent of that controlling response to light intensity, and that both genetic mechanisms are complex.     

Natural entrainment without dawn and dusk: the case of the European ground squirrel (Spermophilus citellus).

Observational data collected in the field and in enclosures show that diurnal, burrow-dwelling European ground squirrels (Spermophilus citellus) never were above ground during twilight at dawn or at dusk. The animals emerged on average 4.02 h (SD = 0.45) after civil twilight at dawn and retreated in their burrows on average 2.87 h (SD = 0.47) before civil twilight at dusk. Daily patterns of light perceived by these burrowing mammals were measured with light-sensitive radio collar transmitters in an enclosure (the Netherlands) and in the field (Hungary). The observational data are corroborated by the telemetry data, which show clear daily patterns of timing of light perception including light perceived from the burrow entrances. The first light was observed by the animals on average 3.54 h (enclosure, SD = 0.45) and 3.60 h (field, SD = 0.31) after civil twilight at dawn, whereas the final observed light was on average 3.04 h (enclosure, SD = 0.64) and 2.02 h (field, SD = 0.72) before civil twilight at dusk. Thus, the animals do not perceive the rapid natural light-dark (LD) transitions that occur at civil twilight. Instead, they generate their own pattern of exposure to light within the natural LD cycle. The classical phase response model for entrainment by light or dark pulses cannot explain how the circadian system of this species remains entrained to the external, natural LD cycle while the major LD transitions are created by its own behavior.