THE RELATIONSHIP BETWEEN THE PROMOTION OF ELONGATION OF FERN PROTONEMATA BY LIGHT AND GROWTH SUBSTANCES

Germinating spores of the fern Onoclea sensibilis L. were grown in darkness, so that they developed as filaments (protonemata). Brief daily exposure of the filaments to red, far-red or blue light increased the rate of filament elongation. Filament elongation was also promoted by indoleacetic acid. When filament elongation was promoted with both indoleacetic acid and exposure to light, the growth promotions caused by red and far-red light were additive to auxin-induced growth. Blue light promoted elongation only at sub-optimal concentrations of auxin. Elongation induced by guanine was additive to red- and far-red-induced elongation. Gibberellic acid had no effect on elongation under any condition. Blue-light-induced elongation resembled auxin-induced elongation in its requirement for exogenous sucrose and sensitivity to inhibition by parachlorophenoxyisobutyric acid. Red and far-red light were active regardless of the presence or absence of sucrose and promoted elongation at a concentration of parachlorophenoxyisobutyric acid which completely inhibited blue-light-induced elongation.     

Photocontrol of spore germination in the fern Thelypteris kunthii

The light requirement for germination in spores of the fern Thelypteris kunthii (Desv.) Morton was fully satisfied by a long period of continuous red light or partially by intermittent, short periods of red light. Red light-potentiated spore germination was inhibited by brief far-red light irradiation, indicating phytochrome involvement. Repeated exposure of spores to prolonged red and short far-red irradiations, or exposure of red-potentiated spores to far-red light after an extended period in darkness, led to their escape from inhibition of germination by far-red light. Prolonged irradiation of spores with blue light before or after red light treatment partially antagonized the effect of red light.     

BLUE LIGHT IN THE DEVELOPMENT OF FERN GAMETOPHYTES AND ITS INTERACTION WITH FAR-RED AND RED LIGHT

Two effects of blue light on the development of Onoclea sensibilis spores are demonstrated. Brief irradiation promotes the protonemal or filamentous type of growth, and the rate of filament elongation is greater than in darkness. Longer periods of irradiation induce the formation of 2-dimensional prothallia. Blue-light treatments which promote filament elongation interact with elongation-promoting far-red light. Far-red irradiation alone promotes filament elongation to a greater extent than blue light, but a blue-light irradiation, either following or preceding far-red treatment, strongly inhibits the far-red promotion. In darkness, a slow recovery from the blue-light-induced loss of sensitivity to far-red takes place. The recovery may be greatly accelerated by interposing a red-light treatment between blue and far-red irradiation.     

THE EFFECT OF LIGHT ON THE CELLULAR COMPONENTS OF POLARIZED GROWTH IN BEAN INTERNODES

First internodes of light-grown bean seedlings exposed to supplementary red and far-red light and those of dark-grown seedlings were sectioned and studied to determine the effects of irradiation on the cellular components of polarized growth. Cell counts and measurements of epidermis, cortex, and pith are given. Increased length of internodes of far-red-treated plants was caused by both increased rate and increased duration of cell elongation. The effect of far-red light is interpreted as a reversal of the accelerating effect of light upon cell maturation. It is suggested that investigations of the mechanism of the red, far-red response of stems be concerned with the processes involved in cell elongation. In darkness, rate and duration of cell division as well as rate and duration of cell elongation were greater than in any of the irradiated plants, indicating that only part of the photocontrol of stem elongation is mediated through the red, far-red system.     

Light germination ofAnthoceros miyabeanus spores

The effects of light on the spore germination of a hornwort species,Anthoceros miyabeanus Steph., were investigated. Spores of this species were photoblastic, but their sensitivities to light quality were different. Under either continuous white, red or diffused daylight, more than 80% of the spores germinated, but under blue light none or a few of them germinated. Under continuous far-red light or in total darkness, the spores did not germinate at all.Anthoceros spores required red light irradiation for a very long duration, i.e., over 12–24 hr of red light for saturated germination. However, the spore germination showed clear photo-reversibility by repeated irradiation of red and far-red light. The germination pattern clearly varied with the light quality. There were two fundamental patterns; (1) cell mass type in white or blue light: spores divide before germination, and the sporelings divide frequently and form 1–2 rhizoids soon after germination, and (2) germ tube type in red light: spores germinate without cell division, and the single-cell sporelings elongate without cell division and rhizoid formation.     

PHOTOMORPHOGENESIS IN PTERIS VITTATA: I. PHYTOCHROME-MEDIATED SPORE GERMINATION AND BLUE LIGHT INTERACTION

1. Spores of the fern Pteris vittata did not germinate under totaldark conditions, while an exposure of the spores to continuouswhite light brought about germination. The germination was mosteffectively induced by red light and somewhat by green and far-red,but not at all by blue light. The sensitivity of spores to redlight increased and leveled off about 4 days after sowing at27–28. The promoting effect of red light could be broughtabout by a single exposure of low intensity. Far-red light givenimmediately after red light almost completely reversed the redlight effect, and the photoresponse to red and far-red lightwas repeatedly reversible. The photoreversibility was lost duringan intervening darkness between red and far-red irradiations,and 50% of the initial reversibility was lost after about 6hr of darkness at 27–28. These observations suggest thatthe phytochrome system controls the germination of the fernspore.
2. When the imbibed spores were briefly exposed to a low-energyblue light immediately before or after red irradiation, theirgermination was completely inhibited. The blue light-inducedinhibition was never reversed by brief red irradiation givenimmediately after the blue light. The escape reaction of redlight-induced germination as indicated by blue light given aftervarious periods of intervening darkness was also observed, andits rate was very similar to that determined by using far-redlight. Spores exposed to blue light required 3 days' incubationin darkness at 27–28 to recover their sensitivity tored light. The recovery in darkness of this red sensitivitywas temperature-dependent. It is thus suggested that an unknownbluelight absorbing pigment may be involved in the inhibitionof phytochrome-mediated spore germination.

(Received August 21, 1967; )     

Control of the Entry into S Phase by Phytochrome and Blue Light Receptor in the First Cell Cycle of Fern Spores

Phytochrome- and a blue light receptor-dependent pathway antagonisticallyregulate the first mitosis in spores of the fern Adiantum capillus-venerisL. This study focused on determining which phase(s) of the cellcycle is positively regulated by phytochrome and negativelyregulated by a blue light receptor in germinating spores. Incorporationof the radioactivity of ³H-thymidine into the acid-insolublematerial prepared from the spores indicated that phytochromein the PFR form induced the entry into S phase of the firstcell cycle in the spores 20-28 h after irradiation with redlight. Blue light treatment before or after red light treatmenttotally prevented the P_FR-induced DNA synthesis. Brief irradiationwith red, far-red or blue light showed no effects on mitosisif the irradiation was given 28 h after the red light induction,during S and M phases. These results indicate that phytochromeand a blue light receptor regulate the entry into S phase duringthe first cell cycle of fern spores. ( Accepted July 10, 1997)     

<Emphasis Type="Italic">In vitro</Emphasis> culture of <Emphasis Type="Italic">Drynaria fortunei</Emphasis>, a fern species source of Chinese medicine “Gu-Sui-Bu”

This study reports spore germination, early gametophyte development and change in the reproductive phase of Drynaria fortunei, a medicinal fern, in response to changes in pH and light spectra. Germination of D. fortunei spores occurred on a wide range of pH from 3.7 to 9.7. The highest germination (63.3%) occurred on ½ strength Murashige and Skoog basal medium supplemented with 2% sucrose at pH 7.7 under white light condition. Among the different light spectra tested, red, far-red, blue, and white light resulted in 71.3, 42.3, 52.7, and 71.0% spore germination, respectively. There were no morphological differences among gametophytes grown under white and blue light. Elongated or filamentous but multiseriate gametophytes developed under red light, whereas under far-red light gametophytes grew as uniseriate filaments consisting of mostly elongated cells. Different light spectra influenced development of antheridia and archegonia in the gametophytes. Gametophytes gave rise to new gametophytes and developed antheridia and archegonia after they were transferred to culture flasks. After these gametophytes were transferred to plastic tray cells with potting mix of tree fern trunk fiber mix (TFTF mix) and peatmoss the highest number of sporophytes was found. Sporophytes grown in pots developed rhizomes.     

Control of Mitosis by Phytochrome and a Blue-Light Receptor in Fern Spores

The first mitosis in spores of the fern A. capillus-veneris was observed under a microscope equipped with Nomarski optics with irradiation from a safelight at 900 nm, and under a fluorescent microscope after staining with 4[prime],6-diamidino-2-phenylindole. During imbibition the nucleus remained near one corner of each tetrahedron-shaped dormant spore, and asymmetric cell division occurred upon brief irradiation with red light. This red light-induced mitosis was photoreversibly prevented by subsequent brief exposure to far-red light and was photo-irreversibly prevented by brief irradiation with blue light. However, neither far-red nor blue light affected the germination rate when spores were irradiated after the first mitosis. Therefore, the first mitosis in the spores appears to be the crucial step for photoinduction of spore germination. Furthermore, experiments using a microbeam of red or blue light demonstrated that blue light was effective only when exposed to the nucleus, and no specific intracellular photoreceptive site for red light was found in the spores. Therefore, phytochrome in the far-red absorbing form induces the first mitosis in germinating spores but prevents the subsequent mitosis in protonemata, whereas a blue-light receptor prevents the former but induces the latter.     

Photocontrol of growth pattern in a tissue isolated from the gametophytes of bracken fern

A tissue isolated from the gametophytes of bracken fern, Pteridiumaquilinum grew as unbranched filaments in red or green light,or in complete darkness. Irradiation with blue or white lightled to a shift from filamentous to two-dimensional pattern ofgrowth in the tissue. (Received February 13, 1969; )     

Action Spectra for Light-induced Elongation in Fern Protonemata

The action spectrum for promotion of elongation of protonemata of Onoclea sensibilis has peaks at 400–420, 580–600 and 640–660 nm. The largest growth increments at saturating light doses are produced by yellow and far-red light. Elongation induced by yellow and far-red irradiation persists in old as well as young filaments, while red-light promotion is found only in young filaments. The growth promotion caused by yellow light is partially reversed by red light down to the level of growth produced by red irradiation alone. Elongation of rhizoids is under reversible red, far-red control, while yellow light is inactive. A model is proposed and discussed in which the light-sensitive elongation of filaments is accounted for by the presence of three distinct photoreceptors: phytochrome; a pigment absorbing yellow light. P₅₈₀; and a pigment absorbing blue light, P₄₂₀.     

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     

EFFECTS OF RED AND FAR-RED ILLUMINATION ON THE ELONGATION OF FERN PROTONEMATA AND RHIZOIDS

The elongation of fern protonemata is controlled by red andfar-red light in an atypical fashion. Red light promotes theelongation of young plants but inhibits the elongation of olderplants. Far-red light promotes elongation regardless of filamentage, and the maximum promotion by far-red is greater than thepromotion which red light causes in young filaments. The elongationof rhizoids is under typical red, far-red control. Red lightpromotes elongation, and a period of far-red illumination followingred light treatment negates the promotive effects of red light. ¹ Present address of the authors: Dept. of Bacteriology andBotany, Syracuse University, Syracuse, New York, U. S. A. (Received November 5, 1962; )     

Photoregulation of asymmetric cell division followed by rhizoid development in the fern Ceratopteris prothalli

Strap-shaped prothalli of CERATOPTERIS: richardii grown in the dark have an apical meristem, a subapical elongation zone and a basal growth cessation zone [Murata et al. (1997) Plant Cell Physiol. 38: 201]. When the dark-grown prothalli were irradiated with continuous white light, marginal cells of the elongation zone divided asymmetrically, and the resulting smaller cells developed into rhizoids. The asymmetric division was also induced by brief irradiation of red light. The effect of red light was cancelled by subsequent irradiation of far-red light, indicating that the asymmetric division was regulated by phytochrome. Since the response to red light was not observed at 10(1) J m(-2) and saturated at 10(2) J m(-2) and the response is photoreversible by far-red light, the photoresponse was classified as a low-fluence response of phytochrome. Although the asymmetric division was induced by brief irradiation of red light, continuous irradiation of white, blue or red light was necessary to induce rhizoid growth. These results indicate that asymmetric division and subsequent cell growth are independently regulated by light in CERATOPTERIS: prothalli.     

Red light-induced phytochrome relocation into the nucleus in Adiantum capillus-veneris

Phytochromes in seed plants are known to move into nuclei in a red light-dependent manner with or without interacting factors. Here, we show phytochrome relocation to the nuclear region in phytochrome-dependent Adiantum capillus-veneris spore germination by partial spore-irradiation experiments. The nuclear or non-nuclear region of imbibed spores was irradiated with a microbeam of red and/or far-red light and the localization of phytochrome involved in spore germination was estimated from the germination rate. The phytochrome for spore germination existed throughout whole spore under darkness after imbibition, but gradually migrated to the nuclear region following red light irradiation. Intracellular distribution of PHY-GUS fusion proteins expressed in germinated spores by particle bombardment showed the migration of Acphy2, but not Acphy1, into nucleus in a red light-dependent manner, suggesting that Acphy2 is the photoreceptor for fern spore germination.     

PHOTOCONTROL OF THE ORIENTATION OF CELL DIVISION IN ADIANTUM. III. EFFECTS OF METABOLIC INHIBITORS

5-Fluorouracil, 8-azaguanine and ethionine were tested on the orientation of cell division to see whether the two-dimensional development of the fern Adiantum gametophytes was due to newly synthesized protein(s). Using the system in which the orientation of cell division was controlled experimentally by sequential treatment with red light, white light and darkness and by the direction of irradiation, all the inhibitors decreased the rates of cell elongation and cell division of the gametophytes, but did not specifically affect the two-dimensional differentiation at all.     

The Photobiology of Fern Gametophytes: I. THE PHENOMENON OF RED/FAR-RED AND YELLOW/F AR-RED PHOTOREVERSIBILITY

This paper shows that the hypothetical yellow-light-absorbingpigment P₅₈₀ is an unnecessary postulate for describing thephotobiology of fern filaments. The existence of P₅₈₀ was originallypredicted on the basis of action and response spectra that assumedthat filament elongation is the growth parameter subject todirect photocontrol. The present work supports an alternativeconcept, that the cross-sectional area at the base of the apicaldome is the photocontrolled parameter. Far-red irradiation reversesthe effects of both red and yellow light, and dose-responsecurves for yellow light parallel but lag behind the curves forred light. These observations indicate that the responses offern filaments to the entire long wavelength spectral region(yellow to far-red) can be attributed to absorption of lightby phytochrome alone.     

Changes in photosensitive stem growth in intact peas following irradiation

Etiolated pea seedlings given a short red-light pretreatment followed by 30 hr of darkness no longer showed a typical red-light inhibition of internode elongation. The induction of phytochrome-insensitive growth was itself mediated by phytochrome, since far-red light reversed the effect of the short red-light pretreatment. Peas grown in white light showed a similar insensitivity to red light. However, in this instance the phytochrome system exerted some control over internode elongation since far-red light promoted growth slightly, and this effect was red-reversible.     

Hormonal influence on photocontrol of the protandry in the genus Helianthus

Under natural photoperiodic conditions protandry in hermaphrodite disc flowers of sunflower (Helianthus annuus L.) is determined by the different elongation rates of the style and filaments. The elongation of the filament and style starts simultaneously after the daily dark period, but the style growth rate is slower. When plants close to anthesis are exposed to continuous white light (WL) a loss of protandry occurs: the filaments do not grow far enough to extrude the anthers from the corolla. The histological analyses show that the number of filament epidermal cells remains unaltered after organ elongation and that cells respond to photoperiod only by cell expansion. Emasculation does not substantially inhibit filament cell expansion, whereas isolation of the filament or stamen from the corolla suggests that this organ could be the perception site of the filament growth stimulus. In vitro treatments with auxin (indole-3-acetic acid, IAA or alpha-naphthaleneacetic acid, NAA) reverses the inhibition of cell expansion caused by continuous WL, whereas gibberellic acid (GA(3)) at high concentrations reproduces the effect of continuous WL. Experiments carried out on various Helianthus spp. show that all these plants have evolved the same photo- and hormonal-control of the protandry. In experiments in which the light treatments were continued for 24 h, the auxins drastically reduced the inhibiting effect of red light (R) and dichromatic treatments FR (far red)+R, whereas GA(3) repressed filament extension regardless of light quality. As far as auxins are concerned, the response of sunflower filaments does not appear to be connected with the polar transport of the hormone. Moreover, the promoting effect of darkness is not mediated by an increase of endogenous free IAA in disc flowers. However, sunflower filaments manifested a similar temporal pattern of response to the light/dark cycle and to auxin.     

PHYTOCHROME ACTION IN ORYZA SATIVA L.: I. GROWTH RESPONSES OF ETIOLATED COLEOPTILES TO RED, FAR-RED AND BLUE LIGHT

1. Under continuous irradiation, the growth of intact rice coleoptilewas strongly inhibited by red light, and somewhat preventedby blue and far-red light. The inhibitory effect of red lighton coleoptile elongation was caused by a low-energy brief irradiation,and a single exposure of 1.5 kiloergs cm^–2 incidentenergy of red light brought about the 50% inhibition. This photoinhibitionof growth was observed only after the coleoptile had elongatedto about 10 mm or longer. The red light-induced effect was reversedby an immediately following brief exposure to far-red light,and the photoresponses to red and far-red light were repeatedlyreversible. The escape reaction of red lightinduced effect tookplace at a rate so that 50% of the initial reversibility waslost within 9 hr in darkness at 27. The inhibition by bluelight and reversal by far-red irradiation was also achievedrepeatedly with successive treatments of the coleoptiles. Theevidence for a low intensity red far-red reversible controlof coleoptile growth, indicative of control by phytochrome,seems clearly established in etiolated intact seedlings.
2. Incontrast, the elongation of apically excised rice coleoptilesegments was promoted by a brief exposure to red light in 0.02M phosphate buffer, pH 7, and the effect was almost completelynullified by an immediately subsequent exposure to far-red light.It becomes evident that the growth of intact coleoptiles wasinhibited by a exposure to red light, while that of excisedsegments in a buffer was rather promoted by red irradiation.The direction of red light induced responses, either promotiveor inhibitory, depends upon the method of bioassay using intactcoleoptiles or their excised segments.

(Received July 24, 1967; )