Violent Spore Release in Some Fungi Imperfecti

Violent spore discharge in the Hyphomycetes Alternaria tenuis,Memnoniella supsimplex, Curvularia lunata, C. geniculata, Corynesporacassiicola, and Zygophiala jamaicensis occurs after transferfrom a humid to a dry atmosphere. Mechanisms of discharge aresuggested.     

Stimulation of Spore Discharge in Sordaria by Carbon Dioxide

The effect on the rate of spore discharge in Sordaria fimicolaof small concentrations of carbon dioxide has been studied.Substitution of an air-stream containing 0.2-2.5 per cent carbondioxide for air freed from this gas circulating over the peritheciaalways led to a marked increase in the rate of spore discharge.     

Effects of Airspeed and Humidity Changes on Spore Discharge in Sordaria fimicola

An improved experimental system is described for studying therate of spore discharge in Sordaria fimicola subjected to anair-stream of controlled rate and relative humidity. Under nearlysaturated conditions the rate of air-flow does not seem to affectthe rate of discharge. Reduction of the R.H. from 95 per centto 70 per cent or 35 per cent leads to an immediate but temporaryincrease in discharge rate. 70 per cent R.H. seems only to affectactual discharge, but prolonged exposure to 35 per cent R.H.interferes as well with earlier stages in spore development.It is suggested that the immediate effect on discharge is physicalrather than biochemical. There is evidence that the perithecialwall plays some part in the control of spore discharge.     

Spore Discharge in Basidiomycetes: III. Spore Liberation from Narrow Hymenial Tubes

The hymenial tubes of Polyporus betulinus were shown to be approximatelycylindrical,and to have a sporulating hymenium over the greater part oftheir surface. Sporophores were tilted through measured anglesfrom the vertical in the field and in the laboratory, and theeffect on spore liberation was measured by determining the numberof spores liberated from particular tubes in standard time.All tilting caused a reduction in spore liberation. The amountof this reduction was found to compare closely with that predictedfrom calculation of the amount of hymenium lying directly abovethe orifice at various angles of tilt. It is therefore concludedthat liberation from the tube can be accounted for by the initialviolent discharge of the spore and gravitational attractionalone. An addendum corrects an earlier paper (1959) in thisseries. It shows that the variation in density of spore depositscollected from Trametes gibbosa was due to air currents in theapparatus, not to variation in sporulation rate.     

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     

On Light-stimulated Spore Discharge in Sordaria

A rough action-spectrum curve (in the range 400–600 mµ)for light-stimulated spore discharge in Sordaria fimicola iscompared with the absorption-spectrum curve of an ethyl-alcoholextract of the fungus. The two curves show some agreement. Amutant lacking black pigment (probably melanin) is shown torespond to light in its spore discharge as does the normal,but evidence is given that it is more sensitive to light. Itseems that the melanin in the perithecial wall of the normalstrain acts as a neutral filter reducing the effectiveness ofthe incident light.     

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.     

Spore discharge in Cordana musae (Zimm.) H?hnel and Zygosporium oscheoides Mont.

In Cordana musae and Zygosporium oscheovides violent spore dischargeoccurs under conditions of rapidly decreasing vapour-pressure;discharge is not dependent on light. Mechanisms of dischargeare postulated. In C. musae, on drying, water evaporates frominside the conidium and conidiophore, causing a negative pressureto develop in the solutions inside these structures and thewalls to be drawn inwards under tension. The sudden appearanceof a gas bubble in one or both of these structures releasesthe tension, and the resultant jolt causes the conidium to beshot away. In Z. oscheoides similar tension and wall distortiondevelop in a specialized conidiophore cell, the vesicle, onwhich the conidia are poised. On appearance of a gas phase,the vesicle wall springs back to its original form and the conidiaare shot away. The vesicle is likened to a single annulus cellin the fern sporangium. The similarity between discharge inC. musae, Z. oscheoides, and Deightoniella torulosa is brieflydiscussed.     

Autotropism in Fungal Spores

Autotropism was examined in germinating spore pairs of Rhizopusstolonifer, Mucor plumbeus, Trichoderma viride, and Botrytiscinerea. When germinated on agar surfaces the first three speciesexhibited negative autotropism, B. cinerea being neutral inits autotropic behaviour. More pronounced negative autotropismwas shown by the first three species when germinated on a filmof Cellophane applied to an agar surface. Under these conditionsB. cinerea displayed positive autotropism. Spore pairs of R. stolonifer germinated on agar containing cellulosepowder or charcoal showed less negative autotropism than onagar alone. Touching spore pairs of each species showed a markedtendency towards cis-ness, i.e. germ-tubes beginning on thesame side of a line joining the two spore centres, under theculture conditions described, the one exception being the reductionin cis-ness recorded when R. stolonifer was germinated on agarcontaining charcoal. Time courses of germination were determined for single sporesand touching spore pairs of R. stolonifer and M. plumbeus anda significant promotion was obtained in the spore pairs as comparedwith the single spores. Although both these species exhibitmarked negative autotropism there was a strong tendency forthe positive germ-tube, i.e. one beginning more nearly towardsits neighbour, to emerge before the negative germ-tube in thosespore pairs having one germtube positive and the other negativein orientation. Also, in R. stolonifer, the replacement of germination-promotersby germination-inhibitors in filtrates from spore suspensionsas they age is correlated with a change from positive to negativeautotropism in germinating members of (+ –) spore pairs. Possible mechanisms are discussed to account for the observedeffects.     

Spore Discharge in Deightoniella torulosa (Syd.) Ellis

Violent spore discharge in the Hyphomycete Deightoniella torulosais described. On drying, a negative pressure develops in thesolution inside the swollen head of the conidiophore. This causesthe thin wall at the top of the conidiophore to cave inwardsand to be brought into a state of tension. The sudden appearanceof a gas phase in the conidiophore head releases this tension,and the wall rapidly returns to its original rounded shape.The conidium, which is attached to the top of the conidiophore,is thus catapulted away. This appears to be the first recordof a gas phase being associated with violent spore dischargein a fungus.     

Spore Germination in Two Tropical Mosses: Fissidenssp. and Racopilum sp

Optimum germination of spores of a Fissidens species occurredat a relatively higher temperature (30 °C) than the optimumgermination of those of a Racopilum species (25 °C). Subsequentprotonemal growth of the two mosses also showed the same differentialtemperature optima. The high-temperature requirement for germinationof Fissidens spores happens to coincide with the maturationand dispersal of the spores in the dry season, and apparentlyfavours the establishment of new shoots. Fissidens sp, Racopilum sp, tropical mosses, spore germination, temperature adaptation     

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.     

Inhibition of Spore Germination in the Cellular Slime Moulds

The phenomenon of auto-inhibition of spore germination has beendemonstrated in the slime moulds Dictyostelium purpureum, strainno. 2; Dictyostelium discoideum, strain no. 1; Dictyosteliummucoroides, strain no. 2; and Polysphondy-Iium violeceum, strainno. 6. A water-soluble substance present in spores was shownto cause this self-inhibition. The substance was isolated fromeach species studied. Evidence as presented indicates that thesame substance occurs in each species of Dictyostelium, butthat it differs from that found in Pv6. An attempt was alsomade to relate this inhibitory substance to the phenomenon of'differential inhibition' in Dp2, Dd, Dm2, and Pu6. An explanationbased on evidence for differential sensitivity to a common inhibitorof spore germination in Dictyostelium and an inhibition in otherthan the spore stage in Pu6 is presented. Apparent differentialproduction of a common inhibitor in Dictyostelium is also reported.     

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.     

Studies on the Sporogenous Lineage in the Moss Timmiella barbuloides IX. Development of the Tapetum

The ultrastructure of tapetal cells in Timmiela barbuloideswas investigated in relation to events of sporogenesis. Aftertheir establishment both internally and externally to the sporogonialinitials, tapetal cells enlarge and assume a permanently polarizedorganization after completion of meiosis. A large vacuole isformed in the cell region distal to the spore sac, the nucleusbecomes centrally located, and amyloplasts lie in the cytoplasmadjacent to the spore sac. An extensive endomembrane systemdevelops in tapetal cells during the stage of exine depositionin spore tetrads. Sheets of rough endoplasmic reticulum developfirst around the nucleus then also in close proximity to theplasma membrane abutting the spore sac. Concomitantly, interveningdictyosomes produce a variety of vesicles. Unusual structureswith vesicle-like profiles also occur in the inner tapetum cellwalls abutting the spore sac. At the same time most of the starchis lost from the plastids in which grana-fretwork systems develop.A massive secretion of extremely electron-opaque material isassociated with perine deposition onto the free spore surfaces.Degeneration of the tapetal cells during the terminal stagesof spore maturation is marked by distortion of the organelles,increase in vacuolation and the appearance of electron-opaquematerial between the sheets of endoplasmic reticulum.Copyright1994, 1999 Academic Press Bryophytes, endomembrane dynamics, Timmiella, ultrastructure, development, tapetum     

Spore Germination Patterns in the Ferns, Cyathea and Dicksonia

Cell division patterns during germination of spores of Cyatheaaustralis, C. cooperi and Dicksonia antarctica were examinedby light microscopy of sectioned materials and by the scanningelectron microscope. In C. australis and C. cooperi the rhizoidwas traced to a small cell formed by an asymmetric divisionof the spore by a wall parallel to its equatorial plane. Incontrast, the rhizoid was formed by a division of the sporeparallel to its polar axis in D. antarctica. In spores of bothgenera, a second division wall oriented in a plane perpendicularto the first gave rise to the protonemal cell. Certain aspectsof germination described here in spores of Cyathea and Dicksoniaare in conflict with the published accounts of spore germinationin these genera. Cyathea, Dicksonia, spore germination, cell division pattern     

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.     

Microsporidian Spore Discharge and the Transfer of Polaroplast Organelle Membrane into Plasma Membrane

Electron microscopic examinations of Glugea hertwigi and Spraguea lophii spores indicated the presence of a single plasma membrane; however, this membrane remained in the spore during the discharge of the sporoplasm from the spore. Although discharged spores retained the old plasma membrane, the extruded sporoplasms acquired a new plasma membrane. In order to determine where the new plasma membrane came from, we used two fluorescent probes with membrane affinities. The markers were tested on unfired and discharged spores. The probe, N-phenyl-1-naphthylamine (NPN), labeled the polaroplast membrane in addition to the apolar groups in the posterior vacuoles of unfired spores. After spore discharge, NPN label disappeared from the spore ghosts except for a slight fluorescence on residual plasma membranes. Much of the NPN-labeled membrane reappeared after spore discharge on the outer envelope of discharged sporoplasms. The probe chlorotetracycline (CTC) labeled calcium-associated membranes of spore polaroplasts. During spore discharge, the CTC fluorescence shifted from the polaroplast organelle of unfired spores to the outer envelope of discharged sporoplasms. These results indicate that the polaroplast organelle may provide the new plasma membrane for discharged microsporidian sporoplasms.     

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 Centrifugation on Asymmetric Cell Division and Differentiation of Fern Spores

We have investigated the effects of centrifugation on sporepolarity, asymmetric cell division, and rhizoid differentiationin the sensitive fern Onoclea sensibilis L. Centrifugation at10000 g for 30 min produces a random orientation of spores withstratification of the cell contents. After centrifugation atmost early stages of development, the nucleus retains its normalpattern of migration from the centre of the ellipsoidal sporeto the proximal face and then to an end of the spore, withoutregard to the orientation of stratification. This indicatesthat the polarity of the spore is stable to centrifugation.As long as the nucleus migrates to an end of the spore and asymmetriccell division occurs, the small cell differentiates into a rhizoid.The arrangement of large cytoplasmic organelles appears to haveno influence on nuclear migration, asymmetric cell division,or rhizoid differentiation. The only period during developmentwhen centrifugation blocks asymmetric cell division is immediatelypreceding and during mitosis and cytokinesis. Spores centrifugedat this stage do not complete nuclear migration, and symmetriccell division results, with neither cell differentiating intoa rhizoid. The source of the stable polarity of the spore isdiscussed. cell polarity, rhizoid differentiation, centrifugation, Onoclea sensibilis L., sensitive fern, fern spores