An unusual effect of the far-red absorbing form of phytochrome: Photoinhibition of seed germination inBromus sterilis L.

Seeds ofBromus sterilis L. germinated between 80–100% in darkness at 15° C but were inhibited by exposure to white or red light for 8 h per day. Exposure to far-red light resulted in germination similar to, or less than, that of seeds maintained in darkness. Germination is not permanently inhibited by light as seeds attain maximal germination when transferred back to darkness. Germination can be markedly delayed by exposure to a single pulse of red light following 4 h inhibition in darkness. The effect of the red light can be reversed by a single pulse of far-red light indicating that the photoreversible pigment phytochrome is involved in the response. The response ofB. sterilis seeds to light appears to be unique; the far-red-absorbing form of phytochrome (Pfr) actually inhibiting germination.Abbreviations Pr red absorbing form of phytochrome - Pfr far-red absorbing form of phytochrome     

Phytochrome control of rapid nyctinastic movements and membrane permeability in Albizzia julibrissin

Summary The rapid nyctinastic movements of Albizzia julibrissin pinnules are under the control of phytochrome. When given prior to a dark period, red light facilitates and far-red light inhibits pinnule closure in the dark. These light effects are mutually photoreversible. The opening reaction of the pinnules following a dark period is mediated mainly by blue light. The nyctinastic closure response is accompanied by an increased rate of electrolyte efflux from the cut pinna base. This observation, coupled with the fact that the rapid nyctinastic movement is not affected by actinomycin D, supports the view that phytochrome control of the sleep movement is not mediated through effects on RNA metabolism, but rather through changes in membrane permeability.     

Phytochrome-mediated germination control of maize caryopses

Maize caryopses sown in water germinate equally well either in darkness or under any light regime. However, when they are imbibed in mannitol solutions, continuous far-red light proves to be strongly inhibitory on the final germination as compared to darkness. Similar but less pronounced inhibition is also exhibited by continuous red or blue light. Intermittent far-red light can partially substitute for continuous far-red light in inhibiting maize caryopsis germination, and its effect is reversed to the intermittent red light level when red light is given immediately after each far-red illumination. These results are interpreted as a proof of existence and involvement of phytochrome in the germination control of maize caryopses, though its manifestation is realized only under osmotic stress.Abbreviations D darkness - FR far-red - R red - B blue - c-FR, c-R, c-B continuous FR, R, B, resp. - i-FR, i-R intermittent FR, R, resp.     

Effect of far-red and green irradiation on the nyctinastic closure of albizzia leaflets

The nyctinastic closing of Albizzia julibrissin pinnules is delayed by exposure to far-red radiation at 710 and 730 nanometers, with the former more effective than the latter. Far-red radiation at 750 and 770 nanometers has no effect on the process. Red light at 660 nanometers, which by itself has no effect, delayed closure when given before or simultaneously with far-red radiation at 750 or 770 nanometers. Low doses of green light, on the other hand, prevented all far-red radiations from delaying closure when given together with one of them. Effectiveness peaks at 550 nanometers. Green light by itself has no effect on the closing process.

From these and previous results, it is concluded that phytochrome is one of two photoreceptors in the process, that the other photoreceptor is an unknown pigment, and that the unknown photoreceptor requires some prior effect of the far-red-absorbing form of phytochrome before its action. Predictions are made of some of the properties of the unidentified pigment.

     

Phytochrome Levels in Light-Grown Avena Change in Response to End-of-Day Irradiations

The effect of 15-minute end-of-day irradiations on photoreversible phytochrome levels in light-grown oat (Avena sativa L., cv Garry) seedlings was investigated. Oat seedlings were grown in a cycle of 8 hours of natural daylight and 16 hours of complete darkness, from sowing until harvest at day 10. The level of extractable, photoreversible phytochrome per unit fresh weight was 60% higher after end-of-day far-red irradiation than after either end-of-day red irradiation or end-of-day far-red followed by end-of-day red. Seedlings irradiated with end-of-day far-red also exhibited a small but significant increase in shoot height and fresh weight per seedling. Extracts of seedlings given each of these end-of-day treatments were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electroblotted, and immunostained with monoclonal antibodies specific to different phytochromes. Regardless of end-of-day light treatment, phytochrome that is abundant in etiolated tissue was below the limit of detection, indicating that one or more of the phytochromes predominating in green tissue changes in abundance.     

A phytochrome-mediated alteration from stem to rosette growth on chicory root explants in vitro

Chicory root explants (Cichorium intybus L. var. foliosum) of two cultivars, taken before and after hydroponic forcing, were cultured in vitro in complete darkness supplemented with red and far-red light treatments. Using 5 min red light per day, the strong stem elongation occurring in complete darkness was converted to rosette formation. This reaction was reversed to stem elongation (accompanied by leaf formation) adding 15 min far-red light after the red light. Fifteen min far-red light per day alone caused the same reaction as 5 min red/15 min far-red light. Far-red light followed by red light caused rosette formation. In stems, formed under complete darkness in vitro, the presence of phytochrome was shown. No phytochrome was detected in the root fragment itself.Abbreviations R red light - FR far-red light - GA gibberellinic acid - A absorbance - FW fresh weight     

The kinetics of type 1 phytochrome in green,light-grown wheat (Triticum aestivum L.)

The kinetics of type 1 phytochrome were investigated in green, light-grown wheat. Phytochrome was measured by a quantitative sandwich enzyme-linked immunosorbent assay using monoclonal antibodies. The assay was capable of detecting down to 150 pg of phytochrome. In red light, rapid first-order destruction of the far-red-light-absorbing form of phytochrome (Pfr) with a half-life of 15 min was observed. Following white light terminated by red, phytochrome synthesis was delayed in darkness by about 15 h compared to plants given a terminal far-red treatment. Synthesis of the red-light-absorbing form of phytochrome (Pr) was zero-order in these experiments. Phytochrome synthesis in far-red light was approximately equal to synthesis in darkness in wheat although net destruction occurred in light-grown Avena sativa tissues in continuous far-red light, as has been reported for other monocotyledons. In wheat, destruction of Pfr apparently did not occur below a certain threshold level of Pfr or Pfr/total phytochrome. These results are consistent with an involvement of type 1 phytochrome in the photoperiodic control of flowering in wheat and other long-day plants.Abbreviations ELISA enzyme-linked immunosorbent assay - FR far-red light - HIR high-irradiance response - Pfr farred-light-absorbing form of phytochrome - Pr red-light-absorbing form of phytochrome - Ptot total phytochrome (Pr + Pfr) - R red light The authors wish to thank Prof. Daphne Vince-Prue (University of Reading) for many helpful discussions regarding this work. Hugh Carr-Smith was supported by a Science and Engineering Research Council studentship and Chris Plumpton by an Agricultural and Food Research Council (AFRC) studentship. B. Thomas and G. Butcher were supported by the AFRC.     

Interaction of phytohormones and far-red irradiation on the nyctinastic closing of Albizzia julibrissin pinnules

In order for far-red radiation at 760 nm to delay dark closing of Albizzia julibrissin pinnules, red light must be given simultaneously with or just prior to it. Studies have been made to determine whether a phytohormone can replace this red light requirement. Abscisic acid, gibberellin, kinetin, and indole-3-acetic acid have been found to replace the red light. Indole-3-carboxylic acid and a cytokinin antagonist are ineffective. In this hormone and far-red interaction, all hormones are effective at μ M or lower concentrations. The hormones show no interaction with red light at 660 nm. Simultaneous irradiation at 550 nm negates the effect of hormone and far-red interaction in delaying leaflet closing. These results are additional evidence that an unidentified far-red absorbing pigment could be involved with phytochrome in some far-red-mediated processes.     

Evidence for phytochrome involvement in light-mediated stomatal movement in Phaseolus vulgaris L.

Observations made with primary leaves of Phaseolus vulgaris L. demonstrated that phytochrome modulates light-induced stomatal movement. Removal of the far-red-absorbing form of the pigment (Pfr) with far-red (FR) radiation decreased the time required by the stomata to reach maximal opening following a dark-to-light transition; this effect of FR was fully reversible with red. Removal of Pfr with FR also decreased the time required to reach maximal closure following a light-to-dark transition, and the rate of closure was dependent on the final irradiation treatment before darkness. No evidence was found for phytochrome involvement in determining stomatal aperture under constant conditions of either darkness of light.Abbreviations and symbols Chl chlorophyll - D darkness - FR far-red - phytochrome photostationary state - Pfr, Pr FR- and R-absorbing forms of phytochrome, respectively - R red     

Differences in Photoresponse and Phytochrome Spectrophotometry Between Etiolated and De-etiolated Pea Stem Tissue

Morphologically similar pea plants having a 4-fold difference in spectrophoto-metrically detectable phytochrome can be produced by pretreatment of etiolated plants with red light (R) or with red and far-red light combined (RF). A search for response differences which could be ascribed to differences in phytochrome content has resulted only in the establishment of differences due to de-etiolation. Segments of etiolated plants differ from those of plants de-etiolated by R and RF pretreatments in 2 ways. Segments from etiolated plants appear to respond rapidly to the far-red absorbing form of phytochrome (P_FR), while segments from de-etiolated plants do not respond rapidly to P_FR. This statement is based upon 2 observations: (i) the red light induced growth inhibition in segments from etiolated plants rapidly escapes reversibility by far-red light, while with segments from R or RF pretreated plants, the red light effect is fully reversed by subsequent far-red light for up to 2 hr; and (ii) segments from etiolated plants were inhibited to a greater degree than were segments from RF pretreated plants when various photostationary state levels of P_FR were maintained for 30 or 90 min and then removed by photoconversion to P_R. The in vivo nonphotochemical transformation curves of the phytochrome of etiolated and RF pretreated plants appear to differ in 2 related respects: (i) the amount of phytochrome destroyed in de-etiolated tissue is greater than that in etiolated tissue, perhaps as a result of the fact that (ii) the rate and extent of apparent reversion of P_FR to P_R in etiolated tissue is about twice that in de-etiolated tissue.     

Antagonistic action of low-fluence and high-irradiance modes of response of phytochrome on germination and beta-mannanase activity in Datura ferox seeds

Seed germination is often induced by a pulse of red light perceived by phytochrome and cancelled by a subsequent pulse of far-red light. When the pulse of red light is followed by several hours of darkness, a pulse of far-red light is no longer effective and prolonged far-red is necessary to block germination. The aim was to investigate whether the red light pulse and prolonged far-red light act on the same or different processes during germination of Datura ferox seeds. Forty-five hours after the inductive red light pulse, germination could not be blocked by one pulse or six hourly pulses of far-red light, but was significantly reduced by 6 h of continuous far-red light. The pulse of red light increased embryo growth potential and the activities of beta-mannanase and beta-mannosidase extracted from the micropylar region of the endosperm. Continuous far-red light had no effect on embryo growth potential or beta-mannosidase activity, but severely reduced the activity of beta-mannanase. The effect of far-red light had the features of a high-irradiance response of phytochrome. Both germination and beta-mannanase activity were restored by a pulse of red light given after the end of the continuous far-red treatment. It is concluded that the low-fluence response and the high-irradiance response modes of phytochrome have antagonistic effects on seed germination and that the control of beta-mannanase activity is one process where this antagonism is established.     

Photocontrol of stem elongation in light-grown plants of Fuchsia hybrida

Stems of the caulescent long-day plant, Fuchsia hybrida cv Lord Byron, showed 2 types of response to light. In one, internode length was increased by far-red irradiation given at the end of an 8 h photoperiod: the response was no greater with prolonged exposure and was less when the start of far-red was delayed. The effect of far-red was reversible by a subsequent exposure to red light. Internode length was inversely proportional to the P_fr/P ratio established before entry to darkness and there was no evidence for loss of P_fr during a 16 h dark period. The inhibitory effect of P_fr acted at a relatively late stage of internode growth. With the development of successive internodes a second response appeared in which stems lengthened following prolonged daily exposures to red or far-red light, or mixtures of the two, or to brief breaks with red or white light. In these later internodes, a short exposure to far-red near the middle of the night was not reversible by red because red alone promoted elongation at this time. Internode length increased with increase in the daily duration of light and, when light was given throughout an otherwise dark period of 16 h, with increase in illuminance to a saturation value of 200 lx from tungsten lamps. Elongation increased as a linear function of decrease in photostationary state of phytochrome down to P_fr/P0.3; however, internodes were shorter in far-red light than in 25% red/red+far-red. It was concluded that stem length is a net response to two modes of phytochrome action. An inductive effect of P_fr inhibits a late stage in internode expansion, and a phytochrome reaction which operates only in light (and may involve pigment cycling) promotes an early stage of internode development. Stem elongation is thus a function both of the daily duration of light and its red/red+far-red content. The outgrowth of axillary buds was controlled by the first type of phytochrome action only.Abbreviations and symbols FR far red light - R red light - P phytochrome - P_fr phytochrome in the far-red light absorbing form - SD 8 h short days - LDP long-day plant - SDP short-day plant     

Control of mitochondrial activities by phytochrome during greening

Mitochondria isolated from 7-day old darkgrown Avena sativa L. (var. Arnold) laminae given 5 min illumination of red light, followed by varying lengths of darkness up to 3 h, showed at least a twofold increase in the rates of both NADH-dependent oxygen consumption and respiratory chain phosphorylation over those of mitochondria isolated from unilluminated tissue. Similar organelles, isolated from tissue given either far-red or red followed by far-red pretreatment, exhibited rates of both functions of between 25% and 75% below those of the mitochondria from unilluminated tissue. The induction-reversion criteria for phytochrome control of respiration and oxidative phosphorylation were satisfied under all experimental conditions during the greening process.Treatment with continuous far-red light, acting presumably through the high irradiance reaction of phytochrome, served to disengage phytochrome activity from photosynthesis. The stimulation of oxidative phosphorylation still occurred under these conditions, slightly slower but much more prolonged in the absence of ATP from photophosphorylation.Abbreviations BSA bovine serum albumen - DAD diaminodurene - EDTA ethylene-diaminetetra-acetic acid - HEPES N-2-hydroxy-ethyl-piperazine-N-2-ethane-sulphonic acid - P_fr phytochrome in the active form     

Phytochrome action and frost hardening in black spruce seedlings

Black spruce [ Picea mariana (Mill.) B.S.P.] development is sensitive to photoperiod. To date the implication of photoperiod, and especially phytochrome, in the frost hardening process of black spruce has not been fully tested. Two light fluence rates, night interruption of darkness, and red vs far-red radiation treatments were applied to black spruce seedlings, followed by freezing at –6°C. Parallel to the freezing test, growth measurements, bud formation and the xylem water potential estimates of the seedlings were done. While dry weight accumulation depends on the irradiation energy level, bud formation and freezing tolerance are photoperiodically sensitive. Furthermore, bud formation and frost hardening are dependent upon whether phytochrome is in the active form or inactive form, as demonstrated by the positive effect of short days, far-red radiation and the reversal of the red effect by far-red radiation. Also, xylem water potential appears to be influenced by short day and far-red conditioning.     

Reversible phytochrome regulation influenced the severity of ozone-induced visible foliar injuries in Trifolium subterraneum L.

In Trifolium subterraneum, oxidative stress caused by ozone has been shown to result in more severe visible foliar injuries when plants were kept in dim broadband white light during the night (i.e. a long photoperiod) compared to darkness during the night (a short photoperiod). As phytochrome signalling is involved in photoperiod sensing, the effect of night-time red and far-red illumination on the ozone-induced response was studied. T. subterraneum plants were treated with ozone enriched air (70?ppb) for either 1?h for a single day or 6?h for three consecutive days. After the first ozone exposure, plants were separated into six night-time light regimes during the two subsequent nights (10?h?day, 14?h night): (1) darkness, (2) far-red light (FR), (3) a short night-break of red followed by far-red light during an otherwise dark night (R FR), (4) a short night-break of red, far-red and finally red light during an otherwise dark night (R FR R), (5) dim white light (L) and (6) red light (R). The treatments L and R resulted in significantly more severe ozone-induced visible foliar injuries relative to D and FR treatments, indicating a phytochrome-mediated response. The night-breaks resulted in a photoreversible and significantly different ozone response depending on the light quality of the last light interval (R FR or R FR R), supporting a photoreversible (between P_r and P_fr) phytochrome signalling response. Thus, in T. subterraneum, the outcome of oxidative stress due to ozone appears to depend on the photoperiod mediated by the night-time conformation of phytochrome.     

Photocontrol of seed germination in Impatiens wallerana Hook. f.

General characteristics of light sensitivity of Impatients wallerana seeds were investigated. Germination was absolutely dependent on light, irrespective of temperature. High percentages of germination were obtained by exposure to long periods of illumination or, alternatively, to several repeated short irradiations with red light. In this case, responsiveness to light was not altered by increasing either the initial incubation period in darkness or the dark intervals between short exposures. Effects of red light were reversed by far-red light, thus demonstrating the involvement of phytochrome. Evidence was presented for an interactive effect, of unknown physiological nature between red and far-red light on the germination of the seeds.Abbreviations P_r phytochrome, red light absorbing form - Pfr phytochrome far-red absorbing form     

Phytochrome and internode elongation in Chenopodium polyspermum L. the light fluence rate during the day and the end-of-day effect

The elongation of the fourth internode of fully green Chenopodium polyspermum L. is strongly stimulated by far-red light (FR) given at the end of the day. The end-of-day effect is more important when the plants had been cultivated for several days with a main light period of 140 Wm^-2 than with a main light period of 85 Wm^-2. There exists a quantitative relationship between the FR end-of-day effect mediated by phytochrome and the value of the light fluence during the day.Abbreviations D darkness - FR far-red light - HWL white light at 140 Wm^-2 - LWL white light at 85 Wm^-2 - PAR photosynthetically active radiation - R red light - WL white light     

Pulvini as the Photoreceptors in the Phytochrome Effect on Nyctinasty in Albizzia julibrissin

The fact that far-red pretreatment slows the closing response of Albizzia julibrissin pinnules to darkness was used to locate the photoreceptor region for the role of phytochrome in nyctinasty, and to determine whether the effect is localized or translocated. Illumination of pinnule tissue alone induced no response, while illumination of an area as narrow as 1 mm, including only the tertiary pulvini and adjacent portions of rachilla and pinnules, was sufficient for a full response. This suggests that the pulvini themselves, the sites of the response, act as photoreceptors. In experiments with various shielding devices, pinnules on the same rachilla responded independently to local illumination, suggesting the absence of any translocatable effects.     

Interactions of green or red light with blue light on the dark closure of Albizzia pinnules

The interactions of green or red light with blue light on the dark closing of Albizzia julibrissin Durazz. pinnules have been investigated. Irradiations at 430, 450 and 470 nm progressively delay dark closing with increasing photon fluence rates. Red or green light alone has no effect. However, when the blue fluence rate is low, both red and green light interact with it and increase the delaying effect of the blue light. When the blue fluence rate is high, green light interacts with it to negate some of the effectiveness of the blue light, while red light has no effect. This is similar to results obtained previously with far-red light. It is suggested that the same unidentified photoreceptor is operating in both the far-red and blue regions. The results also indicate the presence of a blue-only absorbing photoreceptor whose action is increased by phytochrome.