An Analysis of Spore Discharge in Sordaria

Sordaria fimicola can develop mature perithecia from which sporesare discharged when grown in darkness or in light. Under conditionsof alternating dark and light (12 hrs.: 12 hrs.) each day, sporedischarge is periodic with a low rate during the dark period,succeeded by a gradual rise to a relatively high rate in thelight period followed by a decline before the onset of the nextdark period. There is no trace of an endogenous rhythm. Transferfrom darkness to light always leads to an increase in the rateof discharge, and from light to dark to a decrease. The heightof the peak of discharge rate attained in light following adark period seems to be related to the length of the precedingdark period. Experiments with light of different colours but of roughly thesame energy value show that it is the blue rays that are mainlyeffective. From cultures of filter-paper yeast-extract mediuman orange pigment can be extracted with a maximum absorption,in the visible spectrum, at 470 mµ. It is possible thatthis is important in connexion with the sensitivity of the fungusto blue light.     

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.     

Effect of Light on Spore Discharge of Selected Basidiomycetes

Following entrainment of nine selected species of basidiomyceteswith alternating 12 h light and 12 h dark periods, circadianrhythms with nocturnal peaks of spore discharge were found topersist in continuous light or dark for periods of 4 –9d. However, when light and dark periods were reversed followingentrainment, Piptoporus belulinus. Panellus stipticus, Awiculariaauricula and Dacrymyces deliquescens made immediate readjustmentof their rhythms; Coriolus Dersicolor, Stereum hirsutum andClitocybe nebularis took 24 h to readjust with peaks of sporedischarge synchronized with the new dark time periods, whileGanoderma applanatum and Trametes betulina took 48 h to readjust.These reactions indicate exogenous rhythms Light effects, spore discharge, basidiomycetes     

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.     

Studies in Stomatal Behaviour: IX. PHOTOPERIODIC EFFECTS ON RHYTHMIC PHENOMENA IN XANTHIUM PENNSYLVANICUM

An investigation of the effects of different day-length treatmentson stomatal behaviour in Xanthium pennsylvanicum Wall. has shownthat there are differences in long-and short-day treatmentssimilar to those first reported by Schwabe (1952) for Chrysanthemumand Kalanchoe, viz. stomatal opening towards the end of thenight in short days, but not in long days (short night precededby a period of low intensity illumination). In Xanthium therewere in addition very marked differences in rates of stomatalopening in the morning after different lengths of night. Theseeffects were not persistent, there being an immediate reversalupon a change from long- to shrot-day treatment, or vice versa. Further investigation showed that there was an endogenous rhythmaffecting the stomata in continuous darkness; rate of openingwas slow after very short nights, but became greater with extensionof the night to 14–16 hrs., this being the first ‘peak’of an ‘opening ability’ rhythm. The rhythm diedout rapidly and the second cycle was much reduced in amplitude.The period of the rhythm appeared to be approximately 24 hrs.The phase was set mainly by the time of onset of darkness, butthe duration of the pretreatment with low intensity illuminationwas also important—prolonging this was found to reudcethe time in darkness before the first peak. Each hour of lightof 1,500 lux given before darkness was found to be equivalentot approximately 0.3 hr. to darkness. The predominant effect of the length of the preceding nightwas on the slope of the opening curves rather than on the timeof onset of opening following illumination. The first peak of the opening ability rhythm was often manifestedeven in continuous darkness by a period of ‘night opening’of the stomata. The results are discussed and compared with those of other authorsand attempts are made to relate them to theories of the stomatalmechanism.     

Ascospore discharge in Pleospora herbarutn (Pers.) Rabenh. (Dothideales) stimulated by near-ultraviolet radiation

An isolate of P. herbarum from beet seed failed to discharge ascospores in darkness but did so when exposed to light either continuously or cyclically (12 h light/12 h dark). When colonies with mature asci were subjected to a regime of alternating light and darkness for 54 days at a constant temperature of 20^°C, ascospores were discharged over the entire period. Maximum discharge occurred on the 23rd day; few spores were liberated towards the end of the period. Light-induced spore discharge occurred over a wide temperature range (10–30^°C) with the optimum being approximately 14–23^°C. When light of different wavelengths (300 nm-infrared) was tested, only near-ultraviolet (310–330 nm) radiation stimulated ascospore discharge. Vertical height of ascospore discharge was also determined. When ascospores were trapped above colonies over a range of heights (2–80 mm), most spores were caught at 2 mm; none was caught at heights above 30 mm. The number of spores trapped at 30 mm was only 1.3% of the capture at 2 mm.     

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.     

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.     

Studies in Stomatal Behaviour: X. AN INVESTIGATION OF RESPONSES TO LOW-INTENSITY ILLUMINATION AND TEMPERATURE IN XANTHIUM PENNSYLVANICUM

Two experiments are described in which stomatal sensitivityto low-intensity white light was studied for Xanthium pennsylvanicumWall. In the first experiment a daylength extension for 7, 9, or 15hrs. was given using 10, 40, or 160 lux to shorten a basic 16-hr.night, which was also given at its full length as a tenth treatment.Measurements were made of stomatal opening ability on the morningfollowing the different treatments. With a 15-hr. extensionthere was at all intensities a significant response, shown bya reduced rate of opening in the morning. With a 9-hr. extensionusing 40 or 160 lux, opening ability was reduced, but 9 hrs.of 10 lux was insufficinet to produce a detectable effect. The7-hr. extension was ineffective at all three intensities. In the second experiment stomatal behaviour was observed during20 hrs. of either darkness or 10 lux at four temperatures (15,22, 29, and 36°C.). During 20 hrs. of darkness there wasnight opening at all temperatures, but at lower temperaturesit began sooner and lasted longer. These responses to temperaturedid not fit a simple linear relationship, there being a significantcubic term revealed by non-linear regression analysis. Thiscould be explained if the response was considered in terms ofthe magnitude of the change in temperature (from 25°C.)at the beginning of the experiment; there appeared to be sometemperature compensation over a limited range. in 10 lux, nightopening was suppressed at 29° and 36°, but at 15°it was apparently unaffected by the light; at 22° it wasnot completely suppressed by 10 lux but the time of its occurrencewas delayed. Effects of light and temperature are discussed in relation toan endogenous rhythm in darkness which was previoulsy shownto operate in Xanthium pennsylvanicum (Part IX). It is considered that to explain effects of very low intensitylight it will be necessary to recognize a ‘low intensityresponse’ by stomata, which does not operate via changesin guard-cell carbon dioxide.     

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.     

Relative value of light and food as synchronizers of liver phosphorylase circadian rhythm.

The role played by light and feeding schedules on the circadian rhythm of glycogen content and phosphorylase activity of the liver has been studied. In one experiment, mice were subjected to a regimem of constant darkness during 21 days and compared with mice kept in 12 hrs of light alternating with 12 hrs of darkness. Both groups received food and water ad libitum. Liver glycogen content as well as phosphorylase activity showed, with slight differences, similar circadian variations. In a second experiment, mice under similar lighting conditions (LD 12:12), with water access ad libitum, were divided into two groups; one was offered food ad libitum while the other group recieved food from 0700 to 1800 only. This experiment allowed up to compare two different schedules of food intake; ad libitum, normal schedule (from 1800 to 0600) and reversed schedule (from 0700 to 1800). A complete reversal of the circadian rhythm was observed after 21 days in the group with the reverted feeding schedule. We conclude that food can function as the primary synchronizer in spite of the lighting regimen.     

Factors affecting the activity of alatae of the aphids Myzus persicae (Sulzer) and Brevicoryne brassicae (L.)

Apparatus was designed for testing the frequency of flights of aphids under different conditions of temperature, relative humidity, light and pressure. Young aphids (1–4 days after metamorphosis) flew more often and showed less individual variability than older ones; with all ages activity increased for the first few hours under experimental conditions. Starving increased activity for the first 1–2 hr. Aphids used in experiments on a second day flew less frequently than controls which had remained on the plant the first day. Alate B. brassicae were more active than M. persicae : both species showed alternating periods of activity and quiescence.
At light intensities between 100 and 1000 f.c. there was little difference in flight frequency, but below 100 f.c. activity declined rapidly and apparently ceased with darkness.
Changes in relative humidity temporarily affected flight frequency, a change to a higher humidity retarding, a change to a lower increasing it. After adjusting to the change aphids flew readily at all humidities tested between 50 and 100% with temperatures below 80° F. (26.7°C.). The combination of high humidity and high temperature (90°F. = 32·2°C.) sometimes inhibited flight.
Changes of pressure often increased activity temporarily and flight frequency was greater under fluctuating pressure than under constant pressure.
It is concluded that changes in microclimate in crops are adequate to influence frequency of flight of aphids and consequently the spread of virus diseases.     

Photoinduction of Spore Germination in Marchantia polymorpha L. is Mediated by Photosynthesis

The effects of light on spore germination (protrusion of protonemata)in the liverwort Marchantia polymorpha L. were examined. Sporegermination was found to be light dependent and light irradiationfor 10 h or longer was necessary. Test using specific wavelengthsshowed that the entire spectrum from near UV to red light waseffective, red light being the most effective. Spore germinationcould be induced by intermittent irradiation with 15-min redlight pulses given every 1 or 2 h for 24 h. The effect of intermittentred light was not reversed by subsequent or simultaneous far-redlight irradiation. However, spore germination was inhibitedby the photosynthesis inhibitor DCMU (100 µM). Completeinhibition of spore germination was found when DCMU was givenduring the light period. When DCMU was applied during the darkperiods, only a slight reduction of germination rate was observed.Further, it was found that Chl formed in the spores during imbibitionin darkness. Light sensitivity increased at nearly the samerate as the appearance of Chl. Moreover, spore germination wasinduced in total darkness by the addition of glucose to themedium. These results clearly indicate that photosynthesis mediatesthe photoinduction of spore germination in Marchantia polymorpha. (Received May 13, 1999; Accepted July 14, 1999)     

The Effects of Red, Far-red, and Blue Light on the Geotropic Response of Coleoptiles of Zea mays

The effects of red, far-red, and blue light on the geotropicresponse of excised coleoptiles of Zea mays have been investigated.Seedlings were grown in darkness for 5 or 6 days, exposed tovarious light treatments, and then returned to darkness fordetermination of the geotropic response. The rate of response of the coleoptiles is decreased after theyhave been exposed to red light (620–700 mµ, 560ergs cm^–2sec^–1 for the 24 hrs, but not for the 4hrs, preceding stimulation by gravity. Furthermore, their rateof response is greatly reduced if they are exposed to red lightfor 10 min and then returned to darkness for 20 hrs before geotropicstimulation. At 25° C an interval of 6 to 8 hrs elapses between a 10-minexposure to red light and the first detectable decrease in thegeotropic response of the coleoptile. This interval can be lengthenedby exposing the seedlings to low temperatures (0° to 2°C) after the light treatment but cannot be greatly shortenedby increasing the duration of exposure to red light. Using a standard procedure of exposing 5-day-old etiolated seedlingsto light for various times, replacing them in darkness for 20hrs and then determining the response of the coleoptiles to4 hrs geotropic stimulation, it has been found that: (a) Exposureto red light for 15 sec significantly decreases the geotropiccurvature of the coleoptiles and that further reduction occurson increasing the length of the light treatment to 2 and 5 min.(b) Far-red light has no effect on the geotropic response ofthe coleoptiles but it can completely reverse the effect ofred light. After repeated alternate exposure to red and far-redlight the geotropic response of the coleoptile is determinedby the nature of the last exposure, (c) Complete reversal ofthe effect of red light by far-red radiation only occurs whenexposure to far-red follows immediately after exposure to red.The reversing effect of far-red radiation is reduced if a periodof darkness intervenes between the red and far-red light treatments,and is lost after a dark interval of approximately 2 hrs. The effect of red light on the rate of geotropic response ofthe coleoptiles is independent of their age and length at thetime of excision. Blue light acts in a similar way to red light, but the seedlingsare less sensitive to blue than to red light. Coleoptiles grown throughout in a mixture of continuous, weak,red, and far-red light have a lower rate of geotropic responsethan etiolated coleoptiles.     

Flower Opening in Asiatic Lily is a Rapid Process Controlled by Dark-light Cycling

In commerce, Asiatic lilies are picked in bud, each stem holdingseveral buds. We found flower opening was rapid, taking lessthan 4 h both on the stem and for excised buds. Opening wasalso strongly synchronous. For a 12 h day-night cycle, openingbegan late in the dark period, reaching a mid-point after 11h of darkness. This was equally true of buds that were excisedwhen nearly ready to open, and those with 3–4 d of developmentto complete. Reversing day and night reversed the time of opening,and red light was as effective as white light in providing ‘day’conditions. A 15 min light break during the night did not affectthe opening. Lengthening the night (8, 12, 16 h) and shorteningthe day delayed opening from 9, to 11, to 13 h after the startof darkness, respectively. In continuous light and continuousdark, synchronicity was lost. If opening flowers were held inextended darkness, two phases of opening could be discriminated.In a ‘dark phase’, petals opened to approx. 40°,and anthers remained intact. When such flowers were returnedto light, there was a ‘light phase’, where petalsopened further, became more pigmented and began to recurve,and the anthers dehisced, these events taking only 2–3h. The net result was that flowers became fully open and anthersdehisced approx. 2 h after dawn, regardless of daylength. Copyright2000 Annals of Botany Company Asiatic lily, Lilium hybrid, flower opening, timing, endogenous rhythm, synchronicity     

Studies in Sieve-tube Exudation through Aphid Mouth-parts: The Effects of Light and Girdling

Exudation from the mouth-parts of the willow aphid Tuberolachnussalignus (Gmelin) inserted in the phloem of Salix sp. was studied.Leafy rooted cuttings 80–100 cm. in length were grownin artificial light. Exuding mouth-parts were located on thestem below the crown of leaves and the rates of exudation andconcentration of sucrose in the exudate measured whilst theleaves were alternately illuminated (600–800 f.c.) anddarkened. In darkness the rate of sap and sucrose exudationincreased and the concentration fell, relative to the valuesin the light. Similar effects were produced by girdling thestem just beneath the crown of leaves. These were shown to besecondary effects due to a lessening of transpirational tensionsduring darkness or after girdling. They were eliminated if changesin water tension were avoided. Direct effects of light and girdling on exudation were demonstratedby allowing the leaves to assimilate ¹⁴CO₂ and measuring thespecific activity of the exudate. These changes in radioactivityseen against the constancy in total sugar exudation demonstratea switch in source from leaf to stem. An attempt is made toexplain the pattern of changes in specific activity of the exudateduring light and dark periods in terms of shifts in the locationof contributing sources. The rate of translocation between two aphid colonies situateda measured distance apart was estimated by noting the timeswhen the honeydew from each reached a certain level of radioactivity.A mean figure of 28 cm./hr. was obtained.     

Photomorphogenesis of the Gynophore, Pod and Embryo in Peanut, Arachis hypogaea L.

Darkened excized gynophores ceased to elongate after 8–10days in vitro and started to form a pod. Gynophore elongationwas inhibited to a greater extent in total darkness than underlow irradiance, while pod and embryo growth was stimulated indarkness only. Intact gynophores, enclosed in transparent vials containingglass beads, continued to elongate in both light and darkness.In light the elongating gynophores thickened as they penetratedbetween the glass beads, forming a seedless pod at the bottomof the vials. In the dark the elongating gynophores producedsmall pods in which the seeds had started to grow. Excized gynophores elongated in vitro under continuous whitelight at a rate similar to that of intact exposed gynophores.The rate of elongation in vitro, was lower under continuousblue or red-enriched light, than under white light, and wasfurther reduced under continuous far-red irradiation. Pods didnot form during any of the continuous irradiation treatmentsbut only after transfer to darkness, the largest pods formingafter continuous far-red irradiation. As little as 10 min daily exposure to red or far-red irradiancehad the same effect on gynophore elongation as continuous irradiation.Pods formed only when the daily periods of far-red irradiationwere 30 min or less. Reducing the daily exposures to 2 min decreasedthe time to onset of pod formation from 30 to 16 days. Far-redfollowing red irradiation was effective in inhibiting gynophoreelongation stimulated by red irradiation. Pod formation in red/far-redirradiation was only 50 per cent of that observed in far-redirradiation. The involvement of light in continual gynophoreelongation and in the concomitant inhibition of proembryo growthis discussed. Arachis hypogaea L., peanut, gynophore, photomorphogenesis, embryo development, pod development, proembryo     

GROWTH OF BLADES OF NEREOCYSTIS LUETKEANA (PHAEOPHYTA) IN DARKNESS 1

Short term measurements were made of the relative growth rate of the fast growing portions of blades of Ncreocystis luetkeana (Merl.) Post. et Rupr. during exposure to natural daylight and during prolonged darkness. Growth was only slightly, but significantly, faster during the 12 h of daylight than during the 12 h that included 8 h of darkness. Clearly considerable growth occurred at night. In blades amputated 1 cm beyond the zone measured, growth was slower during both night and day. In continuous darkness growth continued for up to 12 days. It was not influenced by amputation of distal blade tissue but it was increased by severance from the bulb and stipe. A mean volume increase of 50% of the blade tissue was recorded. While the organic content decreased, the drop was half that required to support the increase in volume. There was some evidence against translocation. It is possible that cellular biochemical rearrangements allow a light-independent increase in organic material.     

Effect of light intensity on sporulation of Botryosphaeria ribis

Pycnidia were produced by six of seven isolates ofB. ribis at one or more intensity levels of continuous illumination at 21 °C. Under conditions of alternating light (12 h–27 °C) and darkness (12 h–21 °C) pycnidia formed in cultures of six isolates at three or more intensity levels, while one isolate failed to form pycnidia at any intensity level. Pycnidia did not develop when cultures were incubated in complete darkness. Exposure periods as brief as 2 days under continuous illumination at 21 °C induced pycnidial formation. In alternating light (12 h–27 °C) and darkness (12 h–21 °C), the shortest period of exposure which induced pycnidial formation was 4 days. Continuous illumination at 21 °C favored development of uniloculate pycnidia, while alternating light (12 h–27 °C) and darkness (12 h–21 °C) favored formation of multiloculate pycnidia.Contribution No. 22 from The Botany Section of The Department of Biology, The Pennsylvania State University.     

Stimulation of Spore Discharge by Reduced Humidity in Sordaria

An account is given of experiments on the effect of the humidityof an ambient air-stream on spore discharge in Sordaria fimicola.Change from saturated to dry (35 per cent. R. H.) conditionsinvariably led to a striking increase in the rate of spore discharge,and conversely change from dry to damp conditions produced amarked decrease.