A circadian clock measures photoperiodic time in the male lizard Anolis carolinensis

Summary Photoperiod plays an important role in controlling the annual reproductive cycle of the male lizard Anolis carolinensis. The nature of photoperiodic time measurement in Anolis was investigated by exposing anoles to 3 different kinds of lighting paradigms (resonance, T cycles, and night breaks) to determine if photoperiodic time measurement involves the circadian system. Both the reproductive response and the patterns of entrainment of the activity rhythm were assessed. The results show that the circadian system is involved in photoperiodic time measurement in this species and that a discrete photoinducible phase resides in the latter half of the animals' subjective night. Significantly, the ability of the circadian system to execute photoperiodic time measurement is crucially dependent on the length of the photoperiod. Resonance, T cycle and night break cycles utilizing a photoperiod 10–11 h in duration reveal circadian involvement whereas these same cycles utilizing 6 or 8 h photoperiods do not.Abbreviation CRPP circadian rhythm of photoperiodic sensitivity     

Circadian nature of the photoperiodic clock in Japanese quail

Summary The photoperiodic clock in quail (Coturnix colurnix japonica) is based upon a rhythm of photoinducibility (Øi) but the extent to which this rhythm is circadian remains unclear. Two types of experiment investigated this situation. In the first, gonadectomized quail were adapted to live in periods of darkness by training them on a schedule containing one short day and 3 days of darkness (SD/DD/DD/DD). They were then exposed to a single pulse of 6 or 10 h of light at different times across 3 days of darkness. The photoperiodic response, measured by the increase in LH secretion, showed clear rhythmicity, demonstrating unequivocally the circadian nature of Øi. The second set of experiments employed Nanda-Hamner cycles and varied the length of the photoperiod from 6 to 11 h. Responsiveness in a 36 h or a 60 h cycle was highly dependent upon the length of the photoperiod, something not predicted from theory. For instance, LD 6:30 was not photoperiodically inductive but LD 10:26 was clearly inductive. Close analysis of patterns of LH secretion indicated an unexpected delay before induction occurred and then a rapid rise to a stable level of induction. When LH was measured in every pulse under LD 10:26 there was no evidence that LH levels alternately increased and decreased. This is not consistent with the simplest interpretation of Nanda-Hamner experiments where alternate pulses of light are thought to entrain the rhythm or induce a photoperiodic response by coinciding with Øi. It is concluded that the quail's photoinducible rhythm is indeed based on a circadian rhythm but one that is only weakly self-sustaining. Possibly as a consequence of this, the rhythm's behaviour under abnormal photoperiodic cycles may be rather different from that found in other species and from other circadian rhythms in quail.Abbreviations Øi photoinducible phase - LH luteinizing hormone     

A kinetic analysis of the facultative photoperiodic response in Amaranthus retroflexus L.

The ontogenetic change taking place in the facultative photoperiodic response of A. retroflexus to inductive short-day (SD) conditions was studied by exposing plants to continuous induction after different initial exposures to long-days (LD), and comparing the kinetics of their developmental responses (cumulative number of plants with reproductive apices, flowering stage, and height of the apical dome). As the plants progressed from emergence to autonomous flowering (i.e., in non-inductive conditions), their response to continuous induction became progressively more rapid. Reproductive development was initiated following a progressively shorter lag-phase after the start of induction, but its subsequent rate remained unchanged. Until the onset of reproductive development, the undifferentiated upper part of the shoot apex (apical dome) elongated much more rapidly in SD than in LD. However, in both cases reproductive development was initiated when the apex had elongated to about the same extent, after which its elongation accelerated considerably, but to similar rates in both photoperiods. The data indicate that progress towards reproductive development takes place in inductive (SD), as well as in non-inductive (LD) photoperiods, but one cycle of the latter is as effective as 0.20–0.25 of a cycle of the former. —Plants induced at different stages in ontogeny started to change their subsequent branching pattern (ratio of leafy to leafless branches) as soon as induction was delayed beyond autonomous flowering.Abbreviations LD long-days - SD short-days - RGR Relative Growth Rate     

Inheritance of photoperiod-induced flowering in three photoperiodic lines of Aeschynomene americana L.

Summary Induction of flowering by photoperiod was studied in the parental, F₁, F₂, and reciprocal backcross generations of crosses between three photoperiod-responsive Aeschynomene americana L. lines. Generation means appeared additive. Analysis with Mather and Jinks' scaling tests showed little or no epistasis and indicated that an additive-dominance model was adequate. Partitioning components of variation revealed that nearly all variation was additive genetic with dominance and environmental variation negligible. An additive genetic model with two loci, each with two alleles and all alleles having equal net effect, was tested using Power's partitioning method. Results demonstrated that the model fit the data and that there is a major additive genetic system controlling flowering in these crosses, with minor genetic and environmental influences present. Selection for flowering at a desired day length should be feasible.Accepted by A.R. HallauerFlorida Agricultural Experiment Station, Journal Series No. 9251     

Response of the wasp (<Emphasis Type="Italic">Cotesia glomerata</Emphasis>) to larvae of the large white butterfly (<Emphasis Type="Italic">Pieris brassicae</Emphasis>)

The large white butterfly (Pieris brassicae L) first invaded northernmost Japan from Siberia around 1994, and after a few years, began to expand its range. The wasp, Cotesia glomerata (L) parasitizes larvae of the small white butterfly (Pieris rapae crucivora Boisduval), a usual host in the same geographic area. Some Pieris brassicae larvae in Hokkaido have been parasitized by Cotesia glomerata, but the parasitism rate of Pieris brassicae larvae tends to be lower than that of Pieris rapae. To examine the process of parasitizing Pieris brassicae larvae, we observed how the parasitoid wasp responded to the host larvae on damaged leaves. Cotesia glomerata females tended to avoid Pieris brassicae larvae, and even when female wasps inserted their ovipositors into Pieris brassicae larvae, none laid eggs. The parasitoids obtained from Pieris rapae larvae failed to parasitize Pieris brassicae during the host-acceptance step.     

The photoperiodic clock in blackheaded buntings (Emberiza melanocephala) is mediated by a self-sustaining circadian system

Three experimental protocols were employed to clarify whether the circadian system is involved in photoperiodic time-measurement in the blackheaded bunting, Emberiza melanocephala. In a single-pulse paradigm, one 8-h light pulse was delivered at different times to groups of birds across three days of constant darkness (DD). Photoperiodic induction, as measured by a rise in plasma luteinizing hormone (LH), showed clear circadian rhythmicity. The second experiment examined the LH responses in birds exposed to lighting cycles using a Nanda-Hamner type of protocol and confirmed full photostimulation under 6L:30D. The third experiment measured the time of the first photo-induced rise in LH in birds subjected to 30 h of continuous light following entrainment under short days (6L:18D). This experiment aimed to identify the position of the photoinducible phase ( _i). LH first rose at hour 18 following dawn indicating that _i lies in the middle of the day. Plasma concentrations of melatonin were also measured under 6L:18D and 6L:30D light cycles as another physiological marker of the circadian system in buntings. The pattern of melatonin secretion under these LD cycles showed properties consistent with the driving oscillator being circadian in nature. It is concluded that the circadian pacemaker driving the photoinducible rhythm in blackheaded bunting is strongly self-sustaining and free-runs under constant conditions.     

Interspecific egg load assessment of host plants by Pieris rapae butterflies

Egglaying responses of Pieris rapae L. butterflies to the oviposition deterring pheromone (ODP) of Pieris brassicae L. were studied in the laboratory. Choice experiments with ODP treated leaves and control leaves revealed that females perform a strong preference to lay their eggs on the control leaves. This preference is maintained even when during the experiment the control leaf becomes covered with a large number of conspecific eggs. Choice experiments with cabbage leaves with and without P. rapae eggs seem to indicate the absence of intraspecific egg load assessment of host plants in P. rapae. The deterrent effect of the ODP of P. brassicae to P. rapae females persists for at least 8 days. Behavioural observations suggest olfactory hairs as well as gustatory hairs to be involved in the perception of the ODP but electrophysiological recordings of the various chemoreceptors are necessary to confirm this. Finally the prospects of application of this pheromone/kairomone in cabbage pest control are discussed.

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Appréciation de la charge interspécifique en oeufs sur la plante hôte par Pieris rapae

Résumé La réponse au laboratoire de P. rapae à la phéromone dissuadant la ponte (ODP) de P. brassicae a été étudiée par l'oviposition. Des expériences de choix entre des feuilles traitées à l'ODP et des témoins ont montré que les femelles préfèrent nettement les feuilles témoins. Cette préférence s'est maintenue même quand les feuilles témoins ont été recouvertes d'un grand nombre d'oeufs de P. rapae. Ceci peut indiquer l'absence chez P. rapae d'une évaluation de la charge de ses propres oeufs. L'effet dissuadant du ODP de P. brassicae sur les femelles de P. rapae persiste au moins 8 jours. Les observations comportementales suggèrent que des poils olfactifs aussi bien que des poils gustatifs sont impliqués dans la perception d'ODP mais une confirmation de cette hypothèse par enregistrements électrophysiologiques est nécessaire. Les perspectives d'utilisation de cette phéromone/kairomone dans la lutte contre les insectes du chou sont examinées.

     

Effects of light and photoperiodic conditions on abscisic acid in leaves and roots of Acer pseudoplatanus L.

Roots of Acer pseudoplatanus seedlings grown in liquid nutrient medium contained much lower levels of both free and bound abscisic acid than did leaves. The levels of free abscisic acid were similar in young expanding and of mature leaves, but lower in older senscing leaves. Growing plants under long days or short days did not influence the levels of free and bound abscisic acid in leaves. However, under both long days and short days, levels of bound abscisic acid were lower at the end of the dark period than 8 h later during the light period. Phaseic acid was also detected during the light period but never at the end of the dark period.Abbreviations ABA abscisic acid - PA phaseic acid - SD short day - GLC gas-liquid chromatography - LD long day     

Vertebrate insulin induces diapause termination in Pieris brassicae pupae

Summary Due to its close structural homology with the 4K prothoracicotropic hormone isolated from Bombyx mori, we tested the ability of vertebrate insulin to break pupal diapause in a Lepidopteran, Pieris brassicae. Injection of 5g of bovine insulin in diapausing pupae led to diapause termination and synchronous adult eclosion; the effect of insulin was dose-dependent. Bovine insulin-A chain and B chain injected separately failed to show any biological activity suggesting that the intact structure of the molecule is required. Bovine insulin also promoted adult development of decapitated diapausing animals. We show that insulin triggers a reactivation of the neuroendocrine system leading to a neosynthesis of ecdysone beginning 6 days after treatment. This neosynthesis also occurred in beheaded animals suggesting that insulin stimulates the prothoracic glands without acting via the brain.     

Endokrinologische Untersuchungen an dem Wirt-Parasit-System: Pieris brassicae-Apanteles glomeratus

Die von Apanteles glomeratus L. parasitierten Raupen von Pieris brassicae zeigen in Abhängigkeit des Parasitierungstermins eine deutliche Veränderung des Juvenilhormon (JH)-Titer-Verlaufs während des letzten Larvenstadiums. Dabei tritt ein steiler Anstieg des JH-Gehaltes der Wirtshämolymphe im Zusammenhang mit der Häutung der Parasitenlarven vom 1. zum 2. Larvenstadium auf. Aufgrund von Ligations-experimenten konnte nachgewiesen werden, daß die Parasitenlarven selbst für den erhöhten JH-Titer ihrer Wirtsraupen verantwortlich sind, indem sie während ihrer Häutungsphase anscheinend JH in die Wirtshämolymphe abgeben.Eine durch die Parasitierung gesteigerte Syntheseaktivität läßt sich aus den Befunden histologischer Schnitte der Corpora allata frühparasitierter Raupen nicht feststellen. Dagegen weisen die Prothoraxdrüsen parasitierter Raupen zur Mitte des letzten Stadiums eine deutlich kleinere Querschnittsfläche auf als unparasitierte Tiere. Eine dadurch im Zusammenhang mit dem erhöhten JH-Titer bestehende Beziehung zur Häutungsunfähigkeit parasitierter Pieris-Raupen am Ende des letzten Larvenstadiums wird diskutiert.

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Summary The effects of parasitism by Apanteles glomeratus on the hemolymph juvenile hormone (JH) titers of Pieris brassicae during the last larval instar were determined using the Galleria bioassay.Depending on the time of parasitization, a significant increase of the JH titer could be observed when moulting of the parasites from the first to the second larval instar occurred.As neck-ligatured, parasitized Pieris larvae showed a similar increase of the JH titer at this time, it is concluded that the parasite larvae themselves are responsible for the elevation of the titer by delivering JH during their ecdysis into the host's hemolymph.This is supported by histological results from the corpora allata of parasitized and unparasitized caterpillars at the first and third day of the last larval instar, indicating no differences in its secretory activity. The prothoracid glands of parasitized host larvae, however, appear significantly smaller than those of comparable unparasitized ones in the middle of the last instar. A reduced secretory activity at this time, which is assumed from their decreased size, combined with an elevated JH titer may explain why parasitized larvae fail to moult at the end of their larval development.

     

Behavioral manipulation of host caterpillars by the primary parasitoid wasp Cotesia glomerata (L.) to construct defensive webs against hyperparasitism

Many parasitoids control the behavior of their hosts to achieve more preferable conditions. Decreasing predation pressure is a main aim of host manipulation. Some parasitoids control host behavior to escape from their enemies, whereas others manipulate hosts into constructing defensive structures as barriers against hyperparasitism. Larvae of the parasitoid wasp Cotesia glomerata form cocoon clusters after egression from the parasitized host caterpillar of the butterfly Pieris brassicae. After the egression of parasitoids, the perforated host caterpillar lives for a short period and constructs a silk web that covers the cocoon cluster. We examined whether these silk webs protect C. glomerata cocoons against the hyperparasitoid wasp Trichomalopsis apanteroctena. In cocoon clusters that were not covered by silk webs (bare clusters), only cocoons hidden beneath others avoided hyperparasitism. In covered cocoon clusters, both cocoons hidden beneath others and those with a space between them and the silk web avoided hyperparasitism, whereas cocoons that contacted the silk webs were parasitized. The frequency of cocoons that were hidden beneath others increased with the increasing number of cocoons in a cluster, but the defensive effect of cluster size was thought to be lower than that of silk webs. However, the rate of hyperparasitism did not differ between covered and bare clusters when we allowed the hyperparasitoids to attack the cocoon clusters in an experimental arena. This result was thought to have been caused by low oviposition frequency by these hyperparasitoids. As a result, silk webs did not guard the cocoons from hyperparasitoids in our experiments, but would protect cocoons under high hyperparasitism pressure by forming a space through which the ovipositors could not reach the cocoons.     

Metabolic correlates to glycerol biosynthesis in a freeze-avoiding insect,Epiblema scudderiana

Summary The course of glycerol biosynthesis, initiated by exposure to –4°C, was monitored in larvae of the goldenrod gall moth,Epiblema scudderiana, and accompanying changes in the levels of intermediates of glycolysis, adenylates, glycogen, glucose, fructose-2,6-bisphosphate, and fermentative end products were characterized. Production of cryoprotectant was initiated within 6 h after a switch from +16° to –4°C, with halfmaximal levels reached in 30 h and maximal content, 450–500 mol/g wet weight, achieved after 4 days. Changes in the levels of intermediates of the synthetic pathway within 2 h at –4°C indicated that the regulatory sites involved glycogen phosphorylase, phosphofructokinase, and glycerol-3-phosphatase. A rapid increase in fructose-2,6-bisphosphate, an activator of phosphofructokinase and inhibitor of fructose-1,6-bisphosphatase, appeared to have a role in maintaining flux in the direction of glycerol biosynthesis. Analysis of metabolite changes as glycerol production slowed suggested that the inhibitory restriction of the regulatory enzymes was slightly out of phase. Inhibition at the glycerol-3-phosphatase locus apparently occurred first and resulted in a build-up of glycolytic intermediates and an overflow accumulation of glucose. Glucose inhibition of phosphorylase, stimulating the conversion of the activea to the inactiveb forms, appears to be the mechanism that shuts off phosphorylase function, counteracting the effects of low temperature that are the basis of the initial enzyme activation. Equivalent experiments carried out under a nitrogen gas atmosphere suggested that the metabolic make-up of the larvae in autumn is one that obligately routes carbohydrate flux through the hexose monophosphate shunt. The consequence of this is that fermentative ATP production during anoxia is linked to the accumulation of large amounts of glycerol as the only means of maintaining redox balance.Abbreviations G6P glucose-6-phosphate - F6P fructose-6-phosphate - F1, 6P fructose-1,6-bisphosphate - F2,6P ₂ fructose-2,6-bisphosphate - G3P grycerol-3-phosphate - DHAP dinydroxyacetonephosphate - GAP glyceraldehyde-3-phosphate - PEP phosphoenolpyruvate - PFK phosphofructokinase - FBPase fructose-1,6-bisphosphatase - PK pyruvate kinase     

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     

Gibberellins and the photoperiodic control of stem elongation in the long-day plant Agrostemma githago L.

Agrostemma githago is a long-day rosette plant in which transfer from short days (SD) to long days (LD) results in rapid stem elongation, following a lag phase of 7–8 d. Application of gibberellin A₂₀ (GA₂₀) stimulated stem elongation in plants under SD, while 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidine-carboxylate methyl chloride (AMO-1618, an inhibitor of GA biosynthesis) inhibited stem elongation in plants exposed to LD. This inhibition of stem elongation by AMO-1618 was overcome by simultaneous application of GA₂₀, indicating that GAs play a role in the photoperiodic control of stem elongation in this species. Endogenous GA-like substances were analyzed using reverse-phase high-performance liquid chromatography and the d-5 corn (Zea mays L.) assay. Three zones with GA-like activity were detected and designated, in order of decreasing polarity, as A, B, and C. A transient, 10-fold increase in the activity of zone B occurred after 8–10 LD, coincident with the transition from lag phase to the phase of rapid stem elongation. After 16 LD the activity in this zone had returned to a level similar to that under SD, even though the plants were elongating rapidly by this time. However, when AMO-1618 was applied to plants after 11 LD, there was a rapid reduction in the rate of stem elongation, indicating that continued GA biosynthesis was necessary following the transient increase in activity of zone B, if stem elongation was to continue under LD. It was concluded that control of stem elongation in A. githago involves more than a simple qualitative or quantitative change in the levels of endogenous GAs, and that photoperiodic induction alters both the sensitivity to GAs and the rate of turnover of endogenous GAs.Abbreviations AMO-1618 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidine-carboxylate methyl chloride - GA(s) gibberellin(s) - LD long day(s) - LDP long-day plant(s) - SD short day(s)     

Selection for high diapause incidence in blow flies (Calliphora vicina) maintained under long days increases the maternal critical daylength: some consequences for the photoperiodic clock

Using a population of Calliphora vicina from southern Scotland (55 degrees N), showing a critical day length for maternal induction of diapause of about 14.5 h per day, strains of flies were selected for a high incidence of larval diapause under long day length (LD 16:8 h). Diapause incidence was raised from under 10% to almost 100% within five or six generations. The selected flies showed an increase in their critical day length to over 16 h per day, and a high incidence of larval diapause under very long photophases. Selected flies, however, showed mean circadian periods for locomotor activity little different from the original stock, or from flies selected for high diapause under LD 12:12 h, and a Nanda-Hamner profile lacking peaks and troughs of diapause incidence at about 24 h intervals. These results are interpreted to show that the photoperiodic regulation of diapause and the control of overt behavioural rhythmicity are 'separate' physiological systems.     

Activity persistence of granulosis viruses [Baculoviridae] in soils in United Kingdom and Denmark

The persistence of granulosis virus infectivity in soil has been investigated. Virus applied to soil was extracted at different times up to 1 year after the original application and the amount of virus was determined using bioassays. In this way the content of infective capsules remained was determined. It was shown that about 99% of the original virus infectivity was lost during 1 year. Together with other investigations this result indicates that leaching, passive lateral movements and degradation accounts for the main part of the loss.      

Changes in gene expression in the leaf of Lolium temulentum L. Ceres during the photoperiodic induction of flowering

Unifoliated plants of Lolium temulentum L. Ceres were induced to flower by a unique 24-h long day (LD) consisting of the extension of the regular 8-h short day (SD) (400 mol photons·m^–2·s^–1, fluorescence + incandescence) with incandescence at 10–15 mol photonsm ^–2·s^–1. The polyadenylated-RNA complement of leaf blade tissues was analysed at 4-h intervals during the photoperiod extension in LD vs. SD, by using two-dimensional polyacrylamide gel electrophoresis to resolve in-vitro-translated products. Of the 991 spots that were analysed, none appeared or disappeared during the inductive cycle, i.e. no qualitative effect of floral induction was detected, at any time. Sixty-eight spots were found whose intensity was influenced by lengthening of the photoperiod; 50 of them, i.e. ca. 5% of the population analysed, were affected before the end of the extension period and were thus potentially related to floral induction. Many of these RNAs were not quantitatively constant during a 24-h cycle in SD. Seven of them oscillated according to the light-on and the light-off signals, among which three seemed to be controlled by phytochrome since their relative amount increased under the standard light conditions but decreased under incandescence even faster than in darkness. The large majority of other RNAs varied with a timing that was not clearly driven by the alternation of light and darkness, indicating that genes related to the biological clock may be especially sensitive to the lengthening of the photoperiod. Furthermore, seven spots were observed that underwent a phase-shift in LD, which consisted, for six of them, of a phase advance of 4–8 h. The steady-state level of CAB mRNA was analysed because the CAB gene family (encoding the chlorophyll a/b-binding proteins of the light-harvesting complexes) is known to be controlled both by the biological clock and phytochrome. In SD, the level was high in the light and low in darkness; the fluctuation was conducted by a circadian rhythm. When plants were exposed to the inductive LD, the peak of mRNA accumulation that was expected according to the endogenous rhythmicity was abolished, possibly because of the change in light quality during the LD extension.Abbreviations CAB chlorophyll a/b-binding proteins of the light-harvesting complexes - 2D two-dimensional - LD(s) longday(s) - LDP(s) long day plant(s) - SD(s) short day(s) - SDP(s) short day plant(s) This work was supported by the University of Liège through the Action de Recherche Concertée (# 88/93-129). Some analyses were performed with the collaboration of Dr. H. Ougham, Institute of Grassland and Environmental Research, Aberystwyth, UK. The authors also want to thank Dr. F. Cremer (Max Planck Institute for Plant Breeding, Köln, Germany) for critical discussion of the results.