Spore discharge in Entomophthora grylli

Entomophthora grylli Fres. has been found on larvae of Bradysia (Sciaridae, Diptera) on wood in Kansas State University greenhouses since November 1967. Infected larvae crawled to an exposed site in the night and by morning spores of E. grylli were being discharged. In the greenhouse and under controlled environments spore discharge showed a marked periodicity; spore discharge occurred in the light with peaks in the first 1–4 h; discharge then gradually declined but extended into the dark. On the second and third days peaks occurred in the light but were progressively smaller. In a greenhouse under fluctuating conditions twin peaks occurred at 10·00 and 22·00; in a growth chamber, at constant 21° C. 90% r.h., alternating 12 h light and dark periods, spore discharge was similar, with maximum numbers about 21/2 h after the onset of the photo-period. In continuous dark and continuous light no endogenous pattern was evident. Temperature range for spore discharge was 2–28·, with optimum at 15·. Decreasing humidities resulted in decreased spore production and cessation of discharge below 40% r.h.     

Light and Spore Discharge in Ascobolus

Spore discharge in Ascobolus crenulatus occurs both in the lightand in the dark. In a 12 h light: 12 h dark daily regime discharge-ratehas peaks in the dark periods, due apparently to light stimulationwith about half a day's interval between stimulus and response. Using a ‘spore clock’ the course of discharge hasbeen followed for a single apothecium on changing from darknessto light. Exposure to light (500 lux) of wave-lengths around400, 440 and 460 mµ immediately causes ’puffing‘,whilst light of longer wave-length (504 and 580 mµ) hasno effect. Change from darkness to white light has no immediateeffect, but there is a delayed stimulation with a marked increasein discharge-rate 10–14 h later. Simultaneous illuminationof an apothecium, which has been in darkness, by blue light(420 mµ, or 440 mµ, 500lux) and yellow light (580mµ, 500 lux) does not result in puffing. The yellow appearsto prevent the blue light from exerting its effects.     

An Endogenous Rhythm of Spore Discharge in Sordaria fimicola

The sporulation process in Sordaria fimicola has been studiedunder controlled conditions, with particular reference to discharge.Discharge occurred with a circadian periodicity for the greaterpart of the sporulation life of the culture, with peaks at about18.00 hours in either light or darkness. The consistency ofthis phenomenon was confirmed by an analysis of previous work.Fluctuations in light intensity, temperature, relative humidity,carbon dioxide concentration, and vibration were eliminatedas far as possible, as causes of this periodicity. Minor fluctuationsin discharge rate with a period of less than 24 h were probablyrandom. The general nature of the periodicity was shown to be consistentwith some aspects of endogenous rhythms in other organisms.However, spore discharge in S. fimicola is very sensitive tolight, and it is readily entrained to discharge out of phasewith the natural period. The rate of development of two stages of spore formation (delimitationand maturation) were determined, and these were related to therate and periodicity of discharge. From the data it was impossibleto establish at which stage of sporulation the periodicity arose.     

Further Observations on Light and Spore Discharge in Certain Pyrenomycetes

A ‘spore-clock’ for studying the hourly rate ofspore discharge over a 24-hour period is described. A numberof the experiments reported in this paper have involved theuse of this apparatus. In Sordaria fimicola there is a distinct positive light-dischargereaction in a dark-conditioned culture, the rate of spore dischargeincreasing steeply to a peak 2–3 hours after brief stimulationby bright light. Although darkening a light-conditioned cultureleads to an immediate decrease in the rate of discharge, thereis no evidence of a delayed negative dark-discharge reaction. In S. verruculosa with a 12-hours light: 12-hours dark dailyreëgime, more spores are discharged in the dark than inthe light periods if the intensity of illumination is low. Withhigher light intensity there is no significant difference betweenthe number of spores discharged in light and dark periods. Asin S. fimicola there is a positive light-discharge reaction,the interval between stimulus and maximum response being muchlonger (8–12 hours). When a dark-conditioned culture istransferred to light for 48 hours and then returned to darknessfor a further 48 hours it is apparent that not only is therea positive light-discharge reaction but also a negative dark-dischargeresponse. The ‘plateau’ level of discharge is essentiallythe same in light and darkness. It is confirmed that in Hypoxylon fuscum light inhibits discharge.     

Evidence of circadian rhythm of electric discharge in Eigenmannia virescens system

The gymnotid electric fish, Eigenmannia virescens, exhibits electric discharge rhythmicity both in alternate light-dark (LD; 12h light, 12h dark [LD 12:12]) and in constant dark (DD) conditions. It suggests that the electric discharge rhythm is under control of the circadian clock. The free-running periods (FRPs) of electric discharge rhythms at 21°C in DD are greater than, but close to, 24h. The maximum of the electric discharge in the Eigenmannia system peaks approximately at circadian time 6 (CT6) in the middle of the subjective day. The circadian oscillator in the system is temperature compensated. This original report reveals the relationship between electric discharge activity and the circadian pacemaker in Eigenmannia and provides an alternative system to investigate circadian rhythms in vertebrates. (Chronobiology International, 17(1), 43-48, 2000)     

Effect of Changing the Light/Dark Schedule, the Time of Onset of the Light or Dark Period, or the Daylength, on Rhythms of Epidermal Cell Proliferation

Rhythms of labeling and mitotic indices were studied in the hindlimb epidermis of the anuran tadpole Rana pipiens under different light/dark (LD) cycles and daylengths in order to examine the role of the various parameters of the lighting regimen in setting the periods of the rhythms and the timing of the cell proliferation peaks. Altering the time of, or inverting, the 12 h light period on a 24 h day resulted in phase shifting of basically bimodal circadian rhythms with peaks in the light and dark. Thus the cell proliferation rhythms were entrained to the LD cycle. These rhythms also entrained to noncircadian schedules since they lengthened on a 15L : 15D cycle and shortened on a 9L : 9D cycle, although the bimodal characteristic of a peak in the light and a peak in the dark remained. Studies of 18L: 6D and 6L : 18D cycles in which either the time of onset of light or dark was changed relative to the 12L: 12D control indicated that the onset of dark may regulate the timing of the labeling index peaks while the onset of light may determine the time of occurrence of mitotic index peaks. Control of the timing of labeling and mitotic index peaks by different parameters of the LD cycle suggests a mechanism for cell cycle regulation by the environmental lighting schedule. Analysis of the rhythms on all the cycles studied suggested that labeling index rhythms equal the length of, or twice the length of, the dark period. Mitotic index rhythms equal the daylfength or a multiple of the length of the dark period.     

Rhythmic Oscillations in Starch Concentration and Activities of Amylolytic Enzymes and Invertase in Medicago sativa Nodules

Alfalfa (Medicago sativa L.) nodule amylase, starch phosphorylaseand invertase activities and concentrations of starch and proteinwere determined every 4 h for 44 h to determine if daily fluctuationsoccur. Plants experienced 12 h light: 12 h dark (LD) duringentrainment and the first 24 h of the experiment. The last 20h were under continuous darkness (DD). Temperature (21°C)and relative humidity (72%) were always constant. Data wereanalyzed by the cosinor method to determine probabilities ofsinusoidal rhythms with periods between 12.0 and 30.0 h. UnderLD conditions, significant 12.1 h rhythms were found for activitiesof amylase and starch phosphorylase and for starch content.Rhythms in amylase activities and starch content were inverselycorrelated whereas rhythms of starch content and starch phosphorylaseactivity were positively correlated. These data indicate thatnodule starch can be rapidly turned over and that amylases maybe responsible for the degradation. None of these rhythms persistedunder DD conditions. In contrast, invertase activity was rhythmicunder LD (24 h period) and DD conditions (30 h period). No significantrhythmic variations were detected in protein levels throughoutthe entire experiment. (Received August 12, 1985; Accepted November 22, 1985)     

Periodicity of Spore Discharge in Daldinia

In Daldinia concentrica spore discharge under natural conditionsis periodic, most spores being discharged during the night andfew in the daytime. An experimental study has been made of dischargeunder controlled conditions of light and temperature. In continuousdarkness periodic discharge was maintained for 12 days, butthen, although spore output continued, it ceased to be periodic.Return to alternating light (12 hrs., 100 f.c.) and darkness(12 hrs.) at once re-established the periodicity. In continuouslight (100 f.c.) periodic discharge ceased after 2 to 3 days,but was immediately re-established in alternating light (12hrs.) and darkness (12 hrs.). When the fungus was placed underconditions of alternating light and darkness each of 6 hours'duration, two peaks of spore-output were soon developed in the24-hour period. The experiments suggest that the natural periodicityis determined by the alternation of day and night.     

Pathway of dark inorganic carbon fixation in two species of diatoms: influence of light regime and regulator factors on diel variations

Planktonic algae submitted to vertical mixing with a short periodicitycommute many times a day from low to high irradiance levels.To study the influence of this light periodicity, two diatoms,Skeletonema coslatum and Nitzschia turgiduloides, were cultivatedunder alternating conditions of 2 h light/2 h dark (2 h/2 h),simulating vertical mixing in the natural environment. Two otherlight regimes were used: continuous light (CL) and alternatecycles of 12 h light/12 h dark (12 h/12 h). Products synthesizedin the dark by S.costmum during 60 s incubation for 2 h/2 hculture or during 5 min for 12 h/12 h culture were determined.They were essentially sugars, malate, aspartate and glyceratefor 2 h/2 h cells and 12 h/12 h cells taken at the beginningof the light period. In contrast, 12 h/12 h cells taken duringthe darkness or in the middle of the light period and set inthe dark synthesized only amino acids. Our results corroborateprevious reports on dark CO₂ fixation via phosphoenolpyruvatecarboxykinase (PEPCKase, enzyme allowing the fixation of CO₂on PEP and the synthesis of amino acids) with involvement ofa substrate synthesized during the light period, but demonstratethat incorporation also occurs by the C-3 pathway (pathway responsiblefor the major CO₂ fixation in the light) in the very early stagesof the dark period. Another important result highlighted bythis study is the appreciable rate of dark fixation: on average6.7, 8.3 and 12.7% of photosynthesis at saturating photon fluxdensity for N.turgiduloides cultivated under 2 h/2 h, CL and12 h/12 h regime respectively and nearly 12% for S.costatumin the 2 h/2 h light regime. Variation of dark fixation wasinvestigated as a function of hour in the two species. Skeletonemacostatum cells submitted to the 2 h/2 h cycle show a constantrate of light-independent assimilation throughout the day. Bycontrast, both N.turgiduloides grown under the 12 h/12 h or2 h/2 h regime and S.costatum cultured under the 12 h/12 h cycleundergo fluctuations in the rate of dark fixation over the light/darkcycle. The mean dark fixation rate is controlled by the lengthof the photoperiod or the frequency of light fluctuations, dependingon species. We argue that this phenomenon must be taken intoconsideration in primary production calculations. Dependingon whether they are synthesized at the beginning or at the endof the light period, products are somewhat different and therate of fixation varies. This leads us to suggest that the pathwayof dark fixation may be regulated by at least two factors: amountof available substrate and enzyme (RuBPCase and PEPCKase) activityand/or amount.     

Patterns of tetraspore discharge in Caloglossa and Murrayella (Ceramiales, Rhodophyta)

Caloglossa released tetraspores only during dark periods in contrast to a marked light-period release exhibited by Murrayella. Both could be readily converted to a reversed light-dark cycle after a lag period of 1 day. Maximum tetraspore discharge by Caloglossa occurred approximately 4–5 h after the onset of darkness and by Murrayella 4–5 h after the onset of light. Neither 8- nor 16–h light periods affected spore release patterns in Caloglossa, while in Murrayella the spore release pattern was briefly disrupted by short day lengths, but recovered after several days. In both genera, the rhythmicity was disrupted in periods of continual darkness or continual light and appeared to be independent of light quality; however, minimum irradiance levels were critical to maintenance of release patterns. Possible mechanisms of tetraspore discharge periodicity are discussed but since both plants typically have a limited habitat as epiphytes on mangrove trunks and pneumatophores, any speculations as to the adaptive advantages of each pattern tend toward mutual contradiction.     

Circadian Rhythms of Potassium and Sodium Contents in the Growing Front of Neurospora crassa

The temporal changes of potassium (K⁺) and sodium (Na⁺) contentsin the growing front of Neurospora crassa (al-2, bd strain)grown on solid medium showed circadian rhythms which persistedfor at least 45 h in the dark. The K⁺ content reached a maximumat about 10 and 30 h after the transfer from light to darkness,while the Na⁺ content was at a minimum at these times. Boththe rhythms were set off by the light to dark transition andwere not observed in constant light. The phase of the circadianrhythm of conidiation of this strain was delayed by 5 h by exposureto 50 min of white light (photon fluence rate 20.7 W/m²) 7 hafter the light to dark transition. The same exposure significantlychanged the ratio of K⁺ to Na⁺ content in the growing frontmeasured 8 h after the exposure. ³ Present address: Pesticides Research Laboratory, TakarazukaResearch Center, Sumitomo Chemical Co., Ltd., 2-1, 4-chome,Takatsukasa Takarazuka, Hyogo 665, Japan. (Received June 26, 1984; Accepted January 11, 1985)     

Light/dark‐induced effects on behavioral rhythms in suprachiasmatic nucleus‐lesioned rats irrespective of the presence of functional suprachiasmatic nucleus brain implants

Summary

Suprachiasmatic nucleus (SCN)‐lesioned rats which had received a fetal SCN graft were kept in constant red light for three months. After this period it was examined whether those rats that showed a recovered free‐running circadian rhythm could be entrained to light/dark cycles. To this end, they were subjected to a 12 h light/12 h dark schedule, followed by a 12 h light shift and again to dark conditions. In addition, the same regime was imposed on SCN‐grafted rats without recovered circadian rhythms and on sham‐grafted animals with a lesion, which were studied as controls. The presence of an SCN graft was identified immunocytochemically by the presence of vasopressin, vasoactive intestinal polypeptide and somatostatin cells.

Drinking, eating and wheel‐running rhythms were found to synchronize to the light/dark cycles in all rats, not with standing the presence of an SCN graft was. A 12 h light shift was immediately followed by a shift in the three rhythms. Under final dark conditions, free‐running patterns reappeared in rhythm‐recovered animals, without any convincing evidence for entrainment of the rhythms in the pattern of transition.

Behavioral rhythms in SCN‐lesioned rats are apparently masked by 12 h light/dark schedules via other visual pathways than the direct projection from the retina to the SCN.     

The effects of pinealectomy and blinding on the circadian locomotor activity rhythm in the Japanese newt,Cynops pyrrhogaster

We examined the effects of pinealectomy and blinding (bilateral ocular enucleation) on the circadian locomotor activity rhythm in the Japanese newt, Cynops pyrrhogaster. The pinealectomized newts were entrained to a light-dark cycle of 12 h light and 12 h darkness. After transfer to constant darkness they showed residual rhythmicity for at least several days which was gradually disrupted in prolonged constant darkness. Blinded newts were also entrained to a 12 h light/12 h dark cycle. In subsequent constant darkness they showed free-running rhythms of locomotor activity. However, the freerunning periods noticeably increased compared with those observed in the previous period of constant darkness before blinding. In blinded newts entrained to the light/dark cycle the activity rhythms were gradually disrupted after pinealectomy even in the presence of the light/dark cycle. These results suggest that both the pineal and the eyes are involved in the newt's circadian system, and also suggest that the pineal of the newt acts as an extraretinal photoreceptor which mediates the entrainment of the locomotor activity rhythm.Abbreviations circadian period - DD constant darkness - LD cycle, light-dark cycle - LD 12:12 light-dark cycle of 12 h light and 12 h darkness     

Fruiting in Sphaerobolus with Special Reference to Light

Sphaerobolus grows and, provided there is sufficient illumination,fruits readily on oatmeal agar or on malt agar. No effect oflight on vegetative growth can be demonstrated. On the maltmedium, increased fruiting occurs with increase of nutrientup to 4 per cent, malt, but at higher concentrations fruitingis not increased and may be retarded. A chemically defined mediumwith starch as the carbon source allows fruiting, but at a lowlevel. Temperature has a profound effect on basidiocarp development;above 25 C. no fruit-bodies are normally formed although vegetativegrowth is approximately optimal at that temperature. For overallfruit-body production at 20 C, light above 100 lux is necessaryand light remains a limiting factor up to about 1, 000 lux.Under continuous light of suitable intensity, fruit-bodies continueto develop and discharge glebal-masses for many weeks. Thereis a distinct periodicity of discharge with (at 20 C.) about12 days between peaks of activity. This corresponds with thetime taken for basidiocarp initiation and development. A number of developmental stages of the basidiocarp are recognized.The final stage, glebal-mass discharge from stellately openedfruit-bodies, is indifferent to light, but all other stagesare stimulated by light. The light intensity for effective stimulationfalls during development and for the penultimate stage an intensityas low as 1 lux is effective. Only light of wave-length below500 mµ is active in overall basidiocarp development. Inthe sensitive region between 400 mµ and 500 mµ,there appear to be peaks of sensitivity around 440 mµand 480 mµ. In alternating light and darkness, simulating natural conditions,glebal-mass discharge occurs in the light periods. With a regimenof 24 hours light and 24 hours of darkness discharge is mainlyin the dark periods.     

TWO LIGHT REACTIONS IN THE PHOTOPERIODIC CONTROL OF FLOWERING OF LEMNA PERPUSILLA AND L. GIBBA

The effects of light quality on the photoperiodic control inthe flowering of a SD duckweed, Lemna perpusilla strain 6746,and a LD duckweed, L. gibba strain G3, were investigated withspecial reference to the interaction between R and B or FR lights. In the diurnal alternation of R or G light and dark periods,L. perpusilla responded as a SDP, but in that of B or FR lightit was almost daylength-indifferent. On the other hand, L. gibbaresponded as a LDP under B, R or FR light, although the criticallight length was altered by the light quality. In the diurnal alternation of R and B or FR light periods containingno dark period, L. perpusilla flowered with the shortening ofthe optimal and critical R light lengths, compared with theplant exposed to that of R light and dark period. The floweringresponse of L. gibba to the R light length showed double peaks,that is, the first peak at the R duration less than 9 hours,and the second at the R duration longer than 9 hours. The firstpeak corresponds to the optimal R light length in L. perpusilla. Under the CL with a mixture of R and B or FR lights, the floweringand frond production were influenced by the intensity ratioof two light given. In both plants, the optimal ratio of B toR or FR to R for the flowering was always greater than thatfor the frond production. It is suggested that the B or FR light interacts with the Rlight in the photoperiodic process in the plants and this interactionbetween the R and B or FR lights should be of importance forobtaining a better understanding of photoperiodism. (Received August 28, 1965; )     

Growth and Dry-weight Distribution in Callistephus chinensis as Influenced by Lighting Treatment

Plants of two cultivars of Callistephus chinensis (Queen ofthe Market and Johannistag) were grown in 8 h of daylight perday with one of the following treatments given during the 16h dark period: (a) darkness—‘uninterrupted night’,(b) I h of light in the middle of the dark period—a ‘nightbreak’, (c) I min of light in every hour of the dark period—‘cycliclighting’, (d) light throughout—‘continuouslight’. The plants receiving uninterrupted dark periods remained compactand rosetted in habit with small leaves, while leaf expansion,stem extension, and flower initiation were promoted in all threeillumination treatments (b, c, d). Although these three treatmentsproduced similar increases in leaf area, continuous light wasthe most effective for the promotion of both stem growth andflower initiation while cyclic lighting was generally more effectivethan a I-h night break. Continuous light also caused more dry matter to be divertedto stems at any given vegetative dry weight and it was shownthat the stem weight ratio of both varieties was correlatedwith stem length.     

Effect of a Light-break on the Growth and Development of Epilobium hirsutum and Lythrum salicaria in Short Photoperiods

Plants of Epilobium hirsutumand Lythrum salicaria were grownin light for 9 h per day with the following adjustments of lightand dark periods. (a) Photoperiod of 9 h and an uninterrupteddark period of 15 h, (b) photoperiod of 8 h and 1 h of lightof the same intensity in the middle of the dark period. Plants of both the species receiving the 9 h continuous photoperiodremained dwarfed in habit. In the 8 + 1 h light-break treatment,Epilobium showed no response but Lythrum responded by a completedevelopmental shift from a short-day to a long-day form. Leafexpansion, stem elongation, dry matter production and flowerinitiation were promoted. In Epilobium, the pattern of total dry weight distribution toleaves, stem and roots remained similar in the two treatmentsbut in Lythrum light-break treatment caused more dry matterto be diverted to stems at the expense of leaves. These changesin morphogenetic pattern of differentiation were accompaniedby physiological changes as the chlorophyll content and therespiration rates were also higher.     

Regulation of Nitrate Reductase in Acetabularia mediterranea

Nitrate reductase (NR) activity has been characterized on cell-freeextracts of the marine unicellular green alga, Acetabulariamediterranea. Reduced methyl viologen and bromophenol blue aswell as NAD(P)H were effective electron donors for the enzyme. NADH-NR activity increased during cell development reachingits maximum level when the cells approached their maximum length.A substantial increase was also seen in enucleated cells. Theenzyme activity was only present in the green parts of the celland became diminished as the stalks bleached in ageing cells.Under a 12 h light: 12 h darkness photoperiod, NADH-NR activityexhibited pronounced daily fluctuations reaching its maximumon the first hour of illumination (this increase was impairedby cycloheximide) and a minimum in the middle of the dark period. In N-starved algal cultures, NR was constitutively present atappreciable levels and new synthesis took place in the presenceof either NO₃ or low concentrations of NH⁺₄ . However, in thepresence of this cation the enzyme remained mostly in its inactiveform, and could be reactivated in vitro with ferrycyanide. Key words: Nitrate reductase, Acetabularia, daily rhythms, nitrate, ammonium     

Vasopressin deficiency provides evidence for separate circadian oscillators of activity and temperature

Vasopressin-containing Long-Evans and vasopressin-deficient Brattleboro rats were maintained in individual cages while telemetered activity (AC) and body temperature (BT) data were collected. Rats were initially exposed to a 12 h/12-h light/dark cycle (photic zeitgeber) and were allowed ad-libitum access to food and water. Daily feeding, care, and handling (nonphotic zeitgebers) occurred at the beginning of the second hour of the dark cycle. After a 14-day habituation period, rats were subjected to continuous light (LL) or dark (DD) and nonphotic cues were presented irregularly. During the habituation period, both strains exhibited clear 24-h circadian rhythms of AC and BT. In LL or DD, photic cues were removed and nonphotic cues were presented irregularly. There was a shift in the rhythm for Long-Evans animals to 26 h for both AC and BT in LL and 24.6 h in DD. Feeding, care, and handling were seen as minor artifact. In Brattleboro rats, although there were robust 26-h and 24.6-h circadian rhythms of AC in the LL and DD, respectively, BT data were inconsistent and showed sporadic fluctuations. In the BT rhythm of Brattleboro animals, strong peaks were associated with feeding, care, and handling times and trough periods were characterized by a dramatic drop in temperature. This experiment demonstrates that AC and BT are controlled by separate oscillators. In addition, the importance of vasopressinergic fibers in the control of circadian rhythms of BT is evidenced by the loss of circadian rhythms in animals lacking these functional fibers when exposed to free-running paradigms where there is no entrainment of photic or nonphotic oscillators.     

Effects of Anaerobiosis and Respiratory Inhibitors on Blue-Light Inhibition of Spore Germination in Pteris vittata

In a fern, Pteris vittata, inhibition by low-energy blue lightof phytochrome-dependent spore germination was counteractedby anerobiosis and respiratory inhibitors, such as KCN and NaN₃.A 50% inhibition of spore germination in a medium containing0.3 mM NaN₃ required about 8 times longer duration of blue lightirradiation compared with the control. The counteracting effectof NaN₃ continued for about 32 hr after withdrawal of the inhibitor.However, NaN₃ neither induced dark germination nor counteractedthe far-red light inhibition of spore germination. Reducingagents and uncouplers were tested and dithionite and arsenateslightly reversed the blue light inhibition of spore germination. (Received December 17, 1981; Accepted July 8, 1982)