Are two photoreceptors involved in the flowering of a long-day plant?

The effect of daylength extension with narrow spectral bands on the flowering of a long-day plant, Brassica campestris L. cv. Ceres, was investigated to obtain clues to the identity of the photoreceptor involved. Extension of a 9 h photoperiod with 5 h of light pulses at various wavelengths resulted in maximal flowering occurring after irradiation at 710 nm, less at 730 nm, and none at 550, 660 and 750 nm. Flowering at 710 and 730 nm was negated by simultaneous exposures at 550 nm, but not at 660 nm. A short preirradiation at 660 nm enabled a following irradiation at 750 nm to induce flowering. This latter induction was prevented by 550 nm irradiation.
Short flashes of light at 710 nm induced flowering that was negated by a following flash at 550 nm but not at 660 nm. The negation by 550 nm radiation was prevented by subsequent flashes at 710 nm, indicating photoreversibility. A flash at 660 nm enabled subsequent light flashes at 750 nm to initiate flowering that was reversed by a following 550 nm flash.
From the results showing the necessity of red and far-red lights, it is proposed that flowering in this long-day plant is due to two photoreceptors - one is phytochrome and the other an unknown pigment with far-red, green photoreversible properties. By using fluence response data, it is deduced that the unidentified photoreceptor has weak absorption bands in the far-red, but has a strong absorption band in the green. Flowering is induced when effects of red light absorbed by phytochrome interact with effects of far-red light absorbed by the unidentified photoreceptor.     

Wavelength Dependence in the Red and Far-Red for Induction of Phytochrome Increase in Mung Bean Hooks

The wavelength dependence for a radiation induced increase of phytochrome in mung bean hooks (Vigna radiata L.), preirradiated with red light, was determined between 640 to 800 nm. Radiation between 640 to 700 nm and 780 to 800 nm had little effect on phytochrome concentration in hooks pretreated with red. Two bands of far-red light, one at 710 nm and the other at 750 to 760 nm, were found to increase phytochrome content about four times. Besides the requirement for a photochemical process, one or more dark processes appear to be necessary for the induction of phytochrome increase.     

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.     

Phytochrome-mediated effects of near-ultraviolet radiation in the induction of flowering in etiolated Lemna paucicostata T-101, a short-day plant

Induction of flowering of etiolated Lemna paucicostata Hegelm. T-101, a short-day plant, was inhibited by far-red (FR) or blue light (BL) applied at the beginning of a 72-h inductive dark period which was followed by two short days. In either case the inhibition was reversed by a subsequent exposure of the plants to near-ultraviolet radiation (NUV), with a peak of effectiveness near 380 nm. Inhibition by BL or FR and its reversion by NUV are repeatable, i.e., NUV is acting in these photoresponses like red light although with much lower effectiveness. Thus, it is considered that NUV acts through phytochrome and no specific BL and NUV photoreceptor is involved in photocontrol of floral induction on this plant.Abbreviations BL blue light - FR far-red light - NUV near ultraviolet radiation - P red-absorbing form of phytochrome - P_fr far-red absorbing form of phytochrome - R red light     

Flowering responses to altered expression of phytochrome in mutants and transgenic lines of Arabidopsis thaliana (L.) Heynh.

The long-day plant Arabidopsis thaliana (L.) Heynh. flowers early in response to brief end-of-day (EOD) exposures to far-red light (FR) following a fluorescent short day of 8 h. FR promotion of flowering was nullified by subsequent brief red light (R) EOD exposure, indicating phytochrome involvement. The EOD response to R or FR is a robust measure of phytochrome action. Along with their wild-type (WT) parents, mutants deficient in either phytochrome A or B responded similarly to the EOD treatments. Thus, neither phytochrome A nor B exclusively regulated flowering, although phytochrome B controlled hypocotyl elongation. Perhaps a third phytochrome species is important for the EOD responses of the mutants and/or their flowering is regulated by the amount of the FR-absorbing form of phytochrome, irrespective of the phytochrome species. Overexpression of phytochrome A or phytochrome B resulted in differing photoperiod and EOD responses among the genotypes. The day-neutral overexpressor of phytochrome A had an EOD response similar to all of the mutants and WTs, whereas R EOD exposure promoted flowering in the overexpressor of phytochrome B and FR EOD exposure inhibited this promotion. The comparisons between relative flowering times and leaf numbers at flowering of the over-expressors and their WTs were not consistent across photoperiods and light treatments, although both phytochromes A and B contributed to regulating flowering of the transgenic plants.     

Floral initiation in Lolium temulentum L.: the role of phytochrome in the responses to red and far-red light

Summary The possibility that phytochrome is involved in the promotion of flowering by far-red light was investigated. The addition of far-red (FR) to a day extension with red (R) light promotes inflorescence initiation in Lolium. A 2-hour interruption with darkness also promoted flowering compared with the uninterrupted red light control; apex length was further increased by a 10-minute FR irradiation given before the 2-hour dark interruption and was decreased by 10-minutes of R light given in the middle: both FR promotion and R inhibition were reversed by R and FR respectively. Apex length increased approximately linearly with increasing duration of dark interruption up to at least 2 1/2 hours. When varying ratios of R:FR light were substituted for a 2-hour dark period, apex length was increasingly depressed as the % R was increased above 25%; no difference between 25% R/75% FR and 100% FR could be detected. Apex length was inversely linearly related to the calculated [Pfr]/[P] ratios above about 40% Pfr.FR promoted flowering when given during a 5-hour interruption of a day extension with R light but, between 0.25 and 0.90 J m² s^-1, there was no effect of intensity of FR; at 0.11 J m^-2 s^-1 apex length was shorter than at 0.25 J m^-2 s^-1 but longer than in darkness. When the duration of FR (from the beginning of a dark interruption of a day extension with R) was varied, apex length increased with increasing duration of FR up to 1 1/4 to 2 hours but further increasing the duration of FR did not promote flowering more.The results implicate phytochrome in the promotion of flowering by FR light. It has been demonstrated that a low [Pfr]/[P] ratio (less than present in 25% R/75% FR) is needed over a relatively long period of time: this explains why a relatively high proportion of FR light must be added to R for several hours in order to give maximum promotion of flowering. It is concluded that, in Lolium, the increased flowering response to FR light is brought about by a reduction of [Pfr]/[P] ratio at the appropriate time, although the possibility that another effect of far-red is also involved has not been rigorously excluded.     

Inhibitory Action of Blue and Far-Red Light in the Flowering of Lemna paucicostata

In a new strain of short-day duckweed (Lemna paucicostata T-101), blue and far-red light-induced inhibition of flowering was investigated. Flowering of this strain failed to be induced under a short-day photoperiod of blue and far-red light, although it responded as a typical short-day plant in red and white light. When the short-day photoperiod of blue or far-red light was terminated by a 15 min red light pulse, flowering recovered completely. This inducing effect of red light was reversed by subsequent exposure to far-red light. Furthermore, it could be demonstrated that 30 min of blue light completely reversed the flowering inductive effect of 5 min red light and vice versa. Evidence is presented suggesting that the inhibitory action of blue and far red light may be due to the lowering of phytochrome P_fr levels below those required to start the dark reactions which lead to flowering. These results are discussed in relation to the time measurement system of photoperiodism.     

Action spectra for C-glucosylflavone accumulation in Hordeum vulgare plumules

Five-day-old etiolated barley plumules contain the C-glucosylflavones saponarin, lutonarin, and lutonarin 3′-methyl ether. When harvested 24 hr after illumination, increased flavonoid levels were essentially linear with increased energies of monochromatic light at seven wavelengths between 450 and 750 nm. Action spectra for saponarin and for a mixture of lutonarin and its 3′-methyl ether were determined between 380 and 760 nm at 6.6 kerg·cm^?2. The saponarin action spectrum showed distinct peaks at 620 and at 660 nm. These two peaks were similar in their photoreversibility when followed by either 6·6 or 34 kerg·cm^?2 of far-red light. Phytochrome is apparently the photoreceptor for the saponarin action spectrum. Lutonarin and its 3′-methyl ether showed peaks at 520 580, 620 and near 660 nm. The 660 nm peak was not photoreversible by 6·6 kerg·cm^?1, but was by 34 kerg·cm_?2, of far-red light. Phytochrome and protochlorophyll are the likely photoreceptors for these 3′-substituted flavonoids.     

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.

     

Chloroplast movement in Mougeotia induced by blue light pulses

The profile-to-face chloroplast movement in the green alga Mougeotia has been induced by strong blue and near-ultraviolet light pulses (6 J m^-2). Simultaneously, strong red or far-red light (10 W m^-2) was applied perpendicularly to the inducing beam. The response was measured photometrically. Against the far-red background the reciprocity law was found to hold for pulse durations varying two orders of magnitude. The action spectrum exhibited a maximum near 450 nm and a distinct increase in near-ultraviolet. The time-course and the spectral dependence of pulse responses of chloroplasts in Mougeotia were similar to those recorded for other plants which are sensitive only to blue. This points to an alternative sensor system active in the short-wavelength region in addition to the phytochrome system.Abbreviations FR far-red light - Pr red absorbing form of phytochrome - Pfr far-red absorbing form of phytochrome - R red light This paper is dedicated to the memory of Professor Jan Zurzycki     

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.     

Photocontrol of petiole elongation in light-grown strawberry plants

Summary The mode of phytochrome control of elongation growth was studied in fully-green strawberry (Fragaria x Ananassa Duch.) plants. Petiole growth showed two distinct types of response to light. In one, the end-of-day response, petioles were lengthened by low-intensity far-red irradiation for 1 h immediately following the 8 h photoperiod. The response was little or no greater with prolonged exposure and less when the start of far-red was delayed. It was already evident in the first leaf to emerge after treatment began. With the development of successive leaves a second, photoperiodic, type of response appeared, in which petioles lengthened following only prolonged exposure to red, far-red, mixtures of the two, or tungsten lighting, all at low levels of intensity. As with the inhibition of flowering in previous experiments, irradiation with red light during the second half of the otherwise long dark period gave the greatest response.Abbreviations and Symbols FR far-red light - HIR high irradiance response - R red light - P_r phytochrome in the red light absorbing form - P_fr phytochrome in the far-red light absorbing form - SDP short-day plant - LDP long-day plant - PAR photosynthetically active radiation     

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     

Blue- and red-light action in photoorientation of chloroplasts in Adiantum protonemata

An action spectrum for the low-fluencerate response of chloroplast movement in protonemata of the fern Adiantum capillus-veneris L. was determined using polarized light vibrating perpendicularly to the protonema axis. The spectrum had several peaks in the blue region around 450 nm and one in the red region at 680 nm, the blue peaks being higher than the red one. The red-light action was suppressed by nonpolarized far-red light given simultaneously or alternately, whereas the bluelight action was not. Chloroplast movement was also induced by a local irradiation with a narrow beam of monochromatic light. A beam of blue light at low energy fluence rates (7.3·10^-3-1.0 W m^-2) caused movement of the chloroplasts to the beam area (positive response), while one at high fluence rates (10 W m^-2 and higher) caused movement to outside of the beam area (negative response). A red beam caused a positive response at fluence rates up to 100 W m^-2, but a negative response at very high fluence rates (230 and 470 W m^-2). When a far-red beam was combined with total background irradiation with red light at fluence rates causing a low-fluence-rate response in whole cells, chloroplasts moved out of the beam area. When blue light was used as background irradiation, however, a narrow far-red beam had no effect on chloroplast distribution. These results indicate that the light-oriented movement of Adiantum chloroplasts is caused by red and blue light, mediated by phytochrome and another, unidentified photoreceptor(s), respectively. This movement depends on a local gradient of the far-red-absorbing form of phytochrome or of a photoexcited blue-light photoreceptor, and it includes positive and negative responses for both red and blue light.Abbreviations BL blue light - FR far-red light - Pfr far-red-absorbing form of phytochrome - Pr red-absorbing form of phytochrome - R red light - UV ultraviolet     

Action spectrum for the effect of day-extensions on flowering and apex elongation in green,light-grown wheat (Triticum aestivum L.)

Fluence-rate response curves for wavelengths from 640 nm to 730 nm were constructed for the day-extension promotion of flowering in green, light-grown, wheat (Triticum aestivum L., cv. Alexandria), a long-day plant. The resultant action spectrum had action maxima at 660 nm and 716 nm and resembles spectra for the high-irradiance reaction (HIR) seen in etiolated plants. Because, the HIR is thought to be controlled by type I pytochrome (that which is most abundant in etiolated tissue) our results indicate the involvement of type I phytochrome in the photomorphogenesis of a light-grown, green plant.Abbreviations Pr red-light-absorbing form of phytochrome - Pfr far-red-light-absorbing form of phytochrome - Ptot total phytochrome level (Pr+Pfr) - HIR high-irradiance reaction - SDP short-day plant(s) - LDP long-day plant(s)     

Photoperiodic Responses of Brassica campestris cv. Ceres

The photoperiodic responses of Brassica campestris L. cv. Ceres were investigated to determine the suitability of this plant for further studies on the spectral require ments for floral initiation. This is a long-day plant, sensitive to one inductive photocycle on the fourth day from germination. The flowering response increased with the length and intensity of a single period of supplementary light used to extend an 8-hour daylength and was greatest at 25°C. Application of nitrates retarded floral initiation by about two days under short day conditions, but did not affect the re sponse to one long day. Gibberellic acid induced earlier floral initiation under short day conditions. The photoperiodic response was little affected by omitting the main light period immediately before or after the supplementary light, as long as the intensity of supplementary light was greater than 5000 lux. Short interruptions (5–10 minutes) of a single 16-hour dark period with high energy red or far-red radiation did not promote flowering. When given continuously during a single 16-hour dark period, far-red radiation was more effective in flower promotion than an equal energy of red.     

GROWTH REGULATION BY ETHYLENE IN FERN GAMETOPHYTES. IV. INVOLVEMENT OF PHOTOSYNTHESIS IN OVERCOMING ETHYLENE INHIBITION OF SPORE GERMINATION

The inhibitory effects of ethylene on spore germination were investigated. In darkness spore germination was completely inhibited by 10 μ1 · 1⁻¹ ethylene. Light partially overcame this inhibition, and the effect of continuous irradiation with white fluorescent light saturated at about 450 μW · cm⁻². Monochromatic red, blue and far-red light were effective in overcoming ethylene inhibition, whereas green was not. Short periodic exposures to red or far-red light were not sufficient to overcome ethylene inhibition. This suggested that phytochrome was not involved. The photosynthetic inhibitor DCMU blocked the effect of light. Infrared gas analysis showed that photosynthesis saturated at about 450 μW · cm⁻² in white light. Red, blue and far-red light were more efficient photosynthetically than green light; DCMU blocked photosynthesis. Normalized curves of photosynthesis and germination vs. light intensity showed a similar dependence on light energy. It was concluded that light appears to overcome the inhibitory effects of ethylene through some process dependent on photosynthesis.     

Interaction of light and temperature on seed germination of Rumex obtusifolius L.

Germination of Rumex obtusifolius L. seeds (nutlets) is low in darkness at 25° C. Germination is stimulated by exposure to 10 min red light (R) and also by a 10-min elevation of temperature to 35° C. A 10-min exposure to far-red light (FR) can reverse the effect of both R (indicating phytochrome control) and 35° C treatment. Fluence-response curves for this reversal of the effect of R and 35° C treatments are quantitatively identical. Treatment for 10 min with light of wavelenght 680, 700, 710 and 730 nm, after R and 35° C treatment, demonstrates that germination induced by 35° C treatment results from increased sensitivity to a pre-existing, active, far-red-absorbing form of phytochrome (Pfr) in the seeds.Abbreviations FR far-red light - P phytochrome - Pr red-absorbing form of P - Pfr far-red-absorbing form of P - R red light     

Control of Flowering of Xanthium pensylvanicum by Red and Far-red Light

A study was made of the effects of various durations, intensities and combinations of red and far-red light interruptions on the flowering responses of Xanthium pensylvanicum Wallr. A dual response to treatments of far-red light was observed. In short dark periods, far-red light alone did not greatly affect flowering but was able to overcome the inhibition of flowering caused by red light. In dark periods longer than 15 hours, far-red inhibited flowering and added to rather than overcame the inhibition by red light. The dark period length required for far-red inhibition remained the same whether far-red was given at the start or at the eighth hour of darkness.

In 48-hour dark periods Xanthium showed 3 responses to additions of red and far-red light breaks: A) response to red light; B) response to far-red light; and C) response to red followed by far-red light. Red light given any time in the first 30 hours of darkness overcame the inhibitory effect of far-red light given at either the start or the eighth hour of darkness. Red light given later than the thirtieth hour did not overcome the far-red effect.

Approximately the same energy of red light was required to overcome the inhibitory effect of far-red at the second hour of darkness as was required to produce maximum red light inhibition at the eighth hour. Although far-red light was most inhibitory when given early in a long dark period, approximately the same energy of far-red light was required to saturate the far-red response at the fourth, eighth and sixteenth hours.

The results are discussed in relation to other reports of far-red inhibition of flowering in short-day plants.

     

The characteristics of light in floral induction in vitro of Cichorium intybus. The possible role of phytochrome

The action of light in the initiation of floral buds in vitro has been studied using monochromatic light qualities on root explants of a long day plant, Cichorium intybus L. cv. Witloof. Red light (660 nm, 0.30 W m^-2) promotes flowering, while far-red (730 nm, 0.31 W m^-2) and irradiation with combined red + far-red (0.20 + 0.41 W m^-2) have no effect. In short day conditions floral response can be obtained in two ways: 1) by interrupting the dark period with 5 brief irradiations of red light (0.45 W m^-2, 12 min) at regular intervals, although these are counteracted by far-red irradiations of equal intensity and duration; 2) by interrupting the long night with 5 h red light applied during the second third of the night, while at the beginning or at the end it is ineffective. Red light efficiency appears to depend on the photosynthetic activity of the tissues, so that flowering increases with increasing intensity of white light and is suppressed if no white light is supplied. The reproductive development is determined by the coordination of proper irradiation conditions with sufficient sensitivity of the perceiving meristematic cells. The period of highest sensitivity to environmental light conditions in the life cycle of a Cichorium root explant occurs between the 8th and the 16th day after the start of the culture. The data strongly suggest that phytochrome is involved in flower induction of Cichorium in vitro.