Light-dark Regime Research Articles

Movement of spermatozoa in the reproductive tract of adult male Spodoptera litura: daily rhythm of sperm descent and the effect of light regime on male reproduction

Sperm production and movement from the fused testes into the male reproductive tract of the common cutworm Spodoptera litura were studied in insects maintained in a 12 h:12 h light dark (LD) regime. Two types of sperm bundles, eupyrene (nucleated) and apyrene (anucleate) were present in the adult testes. Eupyrene bundles constituted about 25% of the total. Descent of spermatozoa from the testes into the upper vas deferens (UVD) first occurred about 24–30 h before adult eclosion. On entering the reproductive tract, eupyrene spermatozoa remained in bundles while apyrene bundles became dissociated before they reached the UVD. Downward movement of both eupyrene and apyrene spermatozoa within the male tract occurred in a daily rhythm. Sperm descent from the testes into the UVD occurred during the early scotophase, followed by their further descent into the seminal vesicle (SV) during the photophase. Spermatozoa remained in the SV for only a short duration, whence sperm quickly passed through the lower vas deferens into the duplex, which acted as the main sperm storage organ until mating was initiated. During mating 80% of sperm left the duplex, but mating did not influence the number of sperm bundles that subsequently descended into the duplex or the rate of their descent. There was no evidence of sperm reflux. Rearing in constant light (LL) and in constant dark (DD) reduced the number of eupyrene sperm present in the testes of adults that emerged in LL and DD compared to controls (LD), although there was no significant effect on the number of apyrene sperm in the testes. The rhythmic pattern of sperm descent was suppressed in both LL and DD regimes, and the number of sperm in the duplex was adversely affected, with a marked impact in LL reared insects. Male longevity, mating behaviour, oviposition and fertility were found to be more severely affected in LL than in DD.

Circadian regulation of visually evoked potentials in the domestic pigeon, Columba livia.

The avian circadian and visual systems are integrally related and together influence many aspects of birds' behavior and physiology. Certainly, light cycles and their visual perception are the major zeitgebers for circadian rhythms, but do circadian rhythms affect vision? To assess whether visual function is regulated on a circadian basis, flash-evoked electroretinograms (ERGs) and vision-evoked potentials (VEPs) from the optic tectum (TeO) were recorded simultaneously in domestic pigeons at different circadian phases in a light-dark regime (LD) and in constant darkness (DD), while feeding activity was measured to determine circadian phase. In both LD and DD, the amplitudes of ERG b-waves were higher during the day than at night and latencies of a- and b-waves were longer at night. The median effective intensity for ERG a-wave was marginally higher during the day than during the night, indicating greater sensitivity at night, but this rhythm did not persist in DD. The amplitudes of TeO VEPs were also greater during the day, and latencies were greater at night in LD and DD. Together, the data indicate that a circadian clock regulates pigeon visual function at several integrative levels.

Regulation of nitrate reductase expression in leaves by nitrate and nitrogen metabolism is completely overridden when sugars fall below a critical level

ABSTRACTThese experiments use Nia30(145), a tobacco nia1nia2 double null mutant transformed with a NIA2 construct, to define when sugar supply plays the dominating role in the regulation of nitrate reductase (NIA) expression. The null alleles of Nia30(145) are transcribed and translated to produce non‐functional NIA transcript and NIA protein, providing an endogenous reporter system to track NIA expression at the transcript and protein level. The re‐introduced NIA2 construct is expressed at low efficiency, providing a background in which the response to changes in sugar status is not complicated by simultaneous changes in the rate of nitrate assimilation and the levels of nitrate and glutamine. In an alternating light–dark regime, Nia30(145) contained high levels of nitrate and low levels of glutamine and other amino acids. This drives constitutive overexpression of NIA. After transfer of Nia30(145) to continuous darkness, nitrate remains high and glutamine low, but the NIA transcript level and NIA protein decreased significantly within 24 h and were undetectable from 48 h onwards. The decrease of the NIA transcript level was fully reversed and the decrease of NIA protein was partly reversed when leaves were detached from the pre‐darkened plants and supplied with sucrose in the dark. The decrease was not reversed by nitrate or cytokinin. The NIA transcript disappeared when the leaf sugar content fell below 4 μmol hexose equivalents g−1 FW, and recovered when sugars rose above 8 μmol hexose equivalents g−1 FW. It is concluded that low sugar represses NIA, completely overriding signals derived from nitrate and nitrogen metabolism.

Growth, ageing and death of a photoautotrophic plant cell culture.

Batch cultures of photoautotrophic cell suspensions of Chenopodium rubrum L., growing in an inorganic medium on CO2 under a daily balanced light-dark regime of 16: 8 h could be maintained for approximately 100 d without subcultivation. The long-lived cultures showed an initial cell division phase of 4 weeks, followed by a stationary phase of another 4 weeks, after which ageing and progressive cell death reduced the number of living cells and the cultures usually expired after another 3-4 weeks. These developmental phases of the cell culture were characterised with respect to photosynthetic performance, dark respiration, content of phytohormones and capacity of cell division. Cell division of the majority of the cells finished in the G1- or G0-phase of the cell cycle, caused by a pronounced decline in the endogenous levels of auxin and cytokinins. Supply of these growth factors to resting cells resulted in resumption of cytokinesis, at least by some of the cells. However, responsiveness to the phytohormones declined during the stationary phase, and subcultivation was no longer possible beyond day 60 when the phases of ageing and death commenced. Ageing was characterised by a further decline in the photosynthetic capacity of the cells, by a climacteric enhancement of dark respiration, but also by a slight increase in the level of IAA and cytokinins concomitant with a decrease in ethylene. Similarities and differences between the development of batch-cultured photoautotrophic cells of C. rubrum and that of a leaf are discussed with respect to using the cell culture as a model for a leaf.

Hydrogen production by Anabaena variabilis PK84 under simulated outdoor conditions.

Hydrogen production by autotrophic, vanadium-grown cells of Anabaena variabilis PK84, a cyanobacterial mutant impaired in the utilization of molecular hydrogen, has been studied under simulated outdoor conditions. The cyanobacterium was cultivated in an automated helical tubular photobioreactor (4.35 L) under air containing 2% CO(2), with alternating 12-h light (36 degrees C) and 12-h dark (14 degrees to 30 degrees C) periods. A. variabilis steadily produced H(2) directly in the photobioreactor during continuous cultivation for 2.5 months. The maximum H(2) production by the continuously aerated culture under light of 332 microE. s(-1). m(-2) was 230 mL per 12-h light period per photobioreactor and was observed at a growth density corresponding to 3.6 to 4.6 microgram Chl a. mL(-1) (1.2 to 1.6 mg dry weight. mL(-1)). Replacement of air with an argon atmosphere enhanced H(2) evolution by a factor of 2. This stimulatory effect was caused mainly by N(2) deprivation in the cell suspension. A short-term decrease of the CO(2) concentration in the air suppressed H(2) evolution. Anoxygenic conditions over the dark periods had a negative effect on H(2) production. The peculiarity of hydrogen production and some physiological characteristics of A. variabilis PK84 during cultivation in the photobioreactor under a light-dark regime are investigated.

Dynamic studies of phloem and xylein flow in fully differentiated plants by fast nuclear-magnetic-resonance microimaging

A fast nuclear-magnetic-resonance imaging method was developed in order to measure simultaneously and quantitatively the water flow velocities in the xylem and the phloem of intact and transpiring plants. Due to technical improvements a temporal resolution of 7 min could be reached and flow measurements could be performed over a time course of 12-30 h. The novel method was applied to the hypocotyl of 35- to 40-day-old, leafy plants of Ricinus communis which were subjected to different light-dark regimes. The results showed that the xylem flow velocities and the xylem volume flow responded immediately to light on-off changes. Upon illumination the flow velocity and the volume flow increased as expected in respect to literature. In contrast, the phloem flow velocity did not change in response to the light-dark regimes. Interestingly, though, the volume flow in the phloem increased during darkness. These findings can be explained by assuming that the conducting area of the phloem becomes enlarged during the dark period due to opening of sieve pores.

Diurnal Oscillations of Gas Production and Effluxes (CO2 and CH4) in Cores from a Peat Bog

Peat cores (15 cm diam X 30 cm deep) from Ellergower Moss, New Galloway, Scotland were kept and monitored at constant temperature (10 ± 0.1ºC) for gas production using a 1.6 mm diam stainless steel probe fitted with a membrane inlet and connected to a quadrupole mass spectrometer. In the headspace, O2, CO2 and CH4 (measured at m/z values 32, 44 and 15 respectively) showed diurnal fluctuations in low-intensity natural daylight and under a light-dark (LD, 12:12) regime. Over the first few cycles O2 and CO2 increased together in the dark and decreased in the light, whereas CH4 showed variations in antiphase with the other two gases. CO2 and CH4 also showed diurnal oscillations at 15 cm depth, but these decreased together in the light whereas argon (m/z = 40) was not varying. A highly-damped free-run of the oscillations in gas concentrations at 15cm depth was evident for only 3 cycles in complete darkness and at constant temperature. This might suggest desynchronization between individual plants with different free-running periods. A hydrocarbon signal (m/z = 26) at 15 cm depth also showed diurnal cycles but out of phase with CO 2 and CH 4. We postulate a circadian control of microbiological activities imposed by the vascular plants (Carex, Eriophorum, Molinia, Calluna, Erica). Under natural conditions the pronounced temperature sensitivity of CO2 and CH4 emission results in entrainment to daily temperature cycles. The amplitudes of the rhythms are greatest when temperature and light intensity changes are most pronounced, i.e. when the fluctuations in environmental factors are most potent as synchronizers (zeitgebers) and as masking factors.

Adult emergence rhythm of the egg-parasitoid Telenomus busseolae

The adult emergence rhythm of Telenomus busseolae, an egg parasitoid of Sesamia nonagrioides, was examined when parasitoids were exposed to different light-dark regimes. Most of the adult parasitoids emerged throughout the whole period of the photoperiodic cycle. Peak male emergence occurred 2–5 hours earlier than that of females. Adult emergence was asynchronous in continuous darkness or light. However, regimes of alternative light and dark phases such as L4:D20, L8:D16, L12:D12, L16:D8 and L20:D4 h generated a population rhythm with a period length of 24 hours. The peak of the emergence activity moves from the scotophase to the middle of the photophase with an increase of the photophase from 4 to 20 h. Rhythmical activity of adults was synchronised within 2 cycles when immature stages of parasitoid grow under continuous light conditions (LL) and then transferred to L12:D12. Moreover, emergence rhythm persisted and continued in a free-run with a period length of less than 24 hours by transferring a rhythmic culture from L12:D12 h to LL or RR (continuous red light) conditions, indicating the existence of a circadian rhythm. The ecological implications of the expression rhythm relate to better survival of the parasitoids.

Regulation of a NAD+ kinase activity isolated from asynchronous cultures of the achlorophyllous ZC mutant of Euglena gracilis.

NAD+ kinase was isolated by chromatography steps from asynchronous cultures of the achlorophyllous ZC mutant of Euglena gracilis. A non Ca(2+)-calmodulin dependent form whose activity was stimulated by EGTA, was selected for its large quantity and high specific activity. Studies of the kinetic parameters revealed two kinds of NAD+ binding site, depending on NAD+ concentrations, and changes induced by EGTA, Ca2+ and Ca(2+)-calmodulin. The search for effectors, soluble (S) and membrane-bound (P), in Euglena gracilis synchronously grown (in a light-dark regime of 12h:12h), and collected at circadian times (CT)--corresponding to the maximum, CT 17, and to the trough, CT 09, of the circadian rhythm of NAD+ kinase activity--was also undertaken by testing the modulations of the kinetic parameters of the prepared NAD+ kinase. The results suggest: (i) structural changes of NAD+ binding sites depending on NAD+ concentrations; (ii) possible binding of the Mg-ATP-2 (or Ca-ATP-2) on the NAD+ sites, because of their common ADP motif; and (iii) different and specific modulations of the kinetic parameters of the two types of NAD+ binding site by the Ca(2+)-calmodulin complex. In addition, the results indicate, in pelletable fractions isolated at CT 09 and CT 17, the presence of two kinds of effector:(i) the first one, possibly Ca2+, which increases the Vmax's while decreasing the binding of NAD+; (ii) the second one, possibly the Ca(2+)-calmodulin complex, which provokes a complete reverse effect. Each of these two effectors seems to be, alternatively and rhythmically (eight circadian hours apart), partially released from the membranes.

Photoregulation of Chloroplast Gene Transcription in the Chromophytic Alga Heterosigma carterae.

Light acts as a complex regulator of cellular development and gene expression in photoautotrophs. Although light signals are highly effective in controlling cellular division and chloroplast biogenesis in the toxic marine alga Heterosigma carterae, their influence on gene expression has not been well characterized. To address this need cultures of H. carterae synchronized by an alternating light-dark regime were sampled through 12 h of light and 12 h of dark to characterize cell division, chloroplast complement, and chloroplast RNA abundance. These studies have identified a unimodal pattern of chloroplast transcriptional activity for a suite of cellular and photosynthetic genes. To determine the alga's response to a change in photoperiod, 12-h light/12-h dark-synchronized cultures were transferred to constant light and then periodically sampled. Although cellular and chloroplast division cycles remained synchronized in constant conditions for 24 h, the transcriptional apparatus responded by increasing ctRNA abundance within 45 min of the change in photoperiod. However, the ability of the alga to mount this rapid transcriptional response was limited to the first 2 h of the putative dark period. Thus, the chloroplast transcriptional apparatus of H. carterae may initiate a rapid, temporally gated response to a change in photoperiod that is independent of ongoing light-entrained cellular and chloroplast division cycles.

In situ observations of stomatal movements in different light-dark regimes: the influence of endogenous rhythmicity and long-term adjustments

In situ observations of stomatal movements in different light-dark regimes: the influence of endogenous rhythmicity and long-term adjustments

A Crypthecodinium cohnii-like dinoflagellate feeding myzocytotically on the unicellular red alga Porphyridium sp.

A Crypthecodinium cohnii-like dinoflagellate was found to prey on the unicellular red alga Porphyridium sp. The cytoplasm of the prey is ingested by myzocytosis within 10–30 s, and the contents of up to 20 Porphyridium cells can be taken up by one dinoflagellate. The feeding tube is retracted after each uptake process. Cytochalasin D disturbs the suction and the retraction of the feeding tube. Feeding behaviour depends on the light–dark regime: the dinoflagellates prey preferentially in the dark period and finish the trophic phase at the beginning of the light period. They then assemble and encyst, and digestion takes place in the light period. Cell division is restricted to the encysted stage. At the end of the light period excystment takes place, combined with defecation. Isogamy as well as anisogamy and nuclear cyclosis were observed. The trophonts have very thin thecal plates and a microtubular basket with two kinds of elongate vesicles. The microtubules of the tubular basket are also found in young cysts, formed after the end of the feeding period. The fine structure of the digestion vacuoles and of the faecal bodies is described.

In situobservations of stomatal movements in different light-dark regimes: the influence of endogenous rhythmicity and long-term adjustments

The technical device for continuous microscopic observations of stomatal movements in a gas exchange chamber using digital image analysis, earlier described by Kappen et al. (1994), was technically improved. By electronic remote control, it is now possible repeatedly to record over a period of several days identical stomata on a 25 x 50 mm leaf area. The responses of individual stomata to various light-dark sequences in the light and in the dark phase were investigated on attached leaves of Vicia faba (L.). The amplitude of stomatal oscillations caused by alternating light was highest on the first day of an experimental series and decreased with repeated application, indicating a long-term adjustment to changing light conditions by decreasing sensitivity to intermittent darkening. In the usual dark phase light-dark sequences had little effect on the stomatal aperture, which remained small. These results were widely convergent with the stomatal conductance calculated from the gas exchange measurements. Very prominent is the role of an endogenous rhythmicity of the stomatal response. It could be demonstrated (1) by autonomous stomatal opening before the light phase started; (2) by a suppression of dark response in the early morning hours; (3) by a decreasing light stimulation in the afternoon; (4) by further increase of aperture for several hours if no light was provided in the light phase. Closely adjacent stomata could show divergent dark-opening movements pointing to an autonomous control mechanism located in the guard cells. The endogenously controlled morning opening provides full assimilation capacity in the usually humid morning hours when transpiratory water loss associated with CO 2 -uptake is comparatively small.

The influence of a permanent magnetic field on the process of adult emergence in Tenebrio molitor

The duration of the last stage of preadult development (pupal stage) was observed in Tenebrio molitor (yellow mealworm beetle) under controlled conditions ( t = 19 °C, RH = 75%, 10 14 h light-dark regime, light intensity approx. 1000 Ix in the presence of a permanent magnetic field of 320 mT maximum induction, as an additional environmental factor, its intensity being approx. 10 000 times higher than the natural, terrestrial one. Several repeated experiments in the April–October period indicate that the process of adult emergence follows the same pattern in control groups of pupae as in the groups exposed to the magnetic field, but the dynamics of metamorphosis are significantly different. Pupal metamorphosis into adults was more rapid in the presence of a magnetic field. An average reduction of the process of adult emergence (eclosion time) was about 14%.

Feed consumption, growth and growth efficiency of rainbow trout (Oncorhynchus mykiss(Walbaum)) fed on diets containing a bacterial single-cell protein

The aim of the present study was to compare the nutritive value of bacterial single-cell protein (BSCP) with that of fishmeal in rainbow trout (Oncorhynchus mykiss (Walbaum)). Four diets were formulated to contain a total of 458 g crude protein/kg of which 0% was from BSCP in diet 1 (BSCP-0), 25% in diet 2 (BSCP-25), 62.5% in diet 3 (BSCP-62.5) and 100% in diet 4 (BSCP-100); the remainder of the protein was from fishmeal. There were two studies: in study 1, duplicate groups of twenty-five fish were fed on one of the four experimental diets at the rate of 20 g/kg body weight per d for 132 d. Feed consumption rates of individual fish were measured using radiography and the overall apparent absorption efficiency for N in each group was measured over a 2-week period. In study 2, N intake, consumption, absorption and accretion were measured for each fish under controlled environmental conditions (12 h: 12 h light-dark regime; 14 degrees). Higher dietary levels of BSCP resulted in significantly higher feed consumption rates but reduced N absorption efficiency and growth rates. However, a diet containing 25% BSCP (75% fishmeal) did not significantly influence growth rates, feed consumption and absorption efficiency compared with a 100% fishmeal diet. The N growth efficiencies were highest in fish fed on the diet containing the highest level of fishmeal and significantly decreased with increasing BSCP content. Construction of N budgets demonstrated that the reduction in growth in fish eating an increasingly larger proportion of BSCP was due to a decrease in N absorption and an increase in the excretion of urea.

Shifts of thermoeffector thresholds in heat-acclimated rats.

1. Heat-acclimated (HA) rats, kept under a 12:12 h light-dark regime, were subjected to an ambient temperature (Ta) of 33 degrees C for 5 h in the last half of the dark phase for 3 weeks. Control rats were kept under a 12:12 h light-dark regime at a constant Ta of 24 degrees C. 2. After the acclimation period, the rats were then placed in a metabolic chamber at a Ta of 26.5 degrees C, and body core temperature was gradually increased and decreased using an intravenous thermode. Thermoeffector thresholds were determined by the hypothalamic temperature (T(hy)) at the onset of warm-induced tail skin vasodilatation and cold-induced thermogenesis measured by oxygen consumption. 3. Each rat was subjected to the experiment twice, once in the first half and once in the last half of the dark phase, on different days in random order. 4. In HA rats, T(hy) at the onset of both skin vasodilatation and thermogenesis were significantly lower in the last half of the dark phase compared with the first half. In control rats, however, there were no such differences between the two halves. 5. The results suggest that in rats acclimated to daily heat exposure, thermoeffector thresholds shift to lower temperatures only during the period of day when the rats had previously been exposed to heat.

Flowering Response of an Intermediate-day Plant, Salsola komarovii Iljin under Different Photoperiodic Conditions

Summary The photoperiodic flowering response of Salsola komarovii Iljin (Chenopodiaceae) was studied. Theplants flowered earliest under an intermediate-day photoperiodic regime of 12-h light and 12-h dark. The flowering response was strongest under these conditions when the response was observed during an early stage of development, whereas the same level of response was obtained even under short-days when observed at a later stage. The nodes on the main stem at which the first flowers were formed were lowest under photoperiodic regimes with dark periods of 12 h and longer. The flowering response was stronger under a prolonged cycle of 12-h light and 16-h dark regime than under the 12-h light and 12-h dark regime. A light-break given at the middle of a 16-h dark period completely inhibited flowering. The results indicate that S. komarovii is an intermediate-day plant and that the intermediate-day plant is a variation of a short-day plant response that has a requirement for a long light period.

Diazotrophic synchronous growth of a marine unicellular cyanobacterium, Synechococcus sp. strain Miami BG 043511, under aerobic and microaerobic/anaerobic conditions

The growth attributes of an aerobic nitrogen-fixing Synechococcus strain, Miami BG 043511, under aerobic and microaerobic/anaerobic conditions were examined using conventional batch and synchronous culture methods. Generation times of this strain, estimated from the increase in cell density under aerobic and anaerobic batch culture conditions, were 19-23 h and 15-19 h at 30 °C, respectively. It seems, therefore, that atmospheric oxygen did not seriously affect diazotrophic growth in this strain. Under a periodic light-dark regime, cells grew synchronously even under microaerobic/anaerobic conditions. When the aerobic culture entered the light period, a peak of photosynthetic activity was followed by a peak of nitrogenase activity. In contrast, a peak of nitrogenase activity preceded a peak of photosynthetic activity under microaerobic/anaerobic conditions. In both cases, however, cell division was observed at or just after the peak of photosynthetic activity. The difference in the timing of the appearance of nitrogenase activity in microaerobic/anaerobic cultures was ascribed to the inability of cells to generate sufficient ATP under anaerobic dark conditions. Periodic changes in cellular carbohydrate content, associated with the periodic appearances of photosynthetic and nitrogen-fixing activities, were observed under both aerobic and microaerobic/anaerobic conditions. Cellular carbohydrate content increased from 10% to 60% of cell dry weight during the phase of photosynthesis under aerobic conditions, while it reached only 40% under microaerobic/anaerobic conditions. The amount of reserve polysaccharides required to support nitrogen fixation was larger in aerobic cultures than in microaerobic/anaerobic cultures.

Cyclic variations of photosynthetic activity under nitrogen fixing conditions in Synchococcus RF-1

When nitrogen fixing cell cultures of Synechococcus RF-1 were subjected to an alternating lightdark regime (12 h:12 h), a cyclic decrease in the photosynthetic oxygen evolution potential was observed during the dark periods. This rhythm of net photosynthesis rate was maintained for at least two days after transition to continuous light. The decrease in net photosynthesis was accompanied by a stimulation of dark respiration. However, the magnitude of oxygen uptake was considerably smaller than the observed decrease in oxygen evolution. The photosynthetic activity of cells taken from the dark period was characterized by (i) a significantly lower quantum yield and (ii) a strong reduction in the light-saturated rate of photosynthesis. Growing the cultures on nitrate or under continuous light completely suppressed this rhythm. Protein synthesis was not necessary for the recovery of the light-saturated rate of photosynthesis during the light period. The cellular content of chlorophyll a and of phycobiliproteins did not vary between light and dark period, indicating that quantitative changes in the composition of the photosynthetic apparatus are not the basis for the observed oscillations. Regulatory modifications of the photosynthetic efficiency are proposed as an adaptation mechanism to adjust the intracellular oxygen concentration to the needs for nitrogenase activity.

Effects of photoperiod on survival, feeding and development of larvae of the red frog crab, Ranina ranina

Ranina ranina zoea was reared from hatching to megalopa under five photoperiods, from 0-h to 24-h light phase per day (0–24 L) with 6-h intervals, to test their effects on the survival rate, intermolt period, growth, external morphology, food consumption and cannibalism. Continuous light had a negative effect on the larvae; however, continuous darkness was advantageous, except during the final instar and metamorphosis. The survival rate to the megalopa stage was significantly lower under 0 and 24 L conditions than under 12 L conditions, but the rate to the final instar under 0 L conditions was similar to that under 6–18 L conditions. The frequency of normal metamorphosis tended to be low under 0 L conditions. The intermolt period was similar in each group, but accumulated zoeal duration was significantly longer under 24 L conditions than in the other groups, at instars VI and VII. The carapace length was smaller with increasing photoperiod at instars III, V and VII. The number of setae on several appendages was similar in each group, but morphogenesis was retarded on the segmentation of several appendages under 24 L conditions, at instar VII. The amount of food consumed daily was significantly lower under 0 L conditions than under 18 and 24 L conditions at instars I, III and VII. Under 0 L condtions little cannibalism occurred, in contrast with those reared under 6–24 L conditions. A light-dark regime is recommended for the rearing of the larvae.