PHYTOCHROME-MEDIATED THRESHOLD CONTROL OF HYPOCOTYL GROWTH IN PARTLY DE-ETIOLATED MUSTARD (Sinapis alba L.) SEEDLINGS

Abstract— Hypocotyl elongation in etiolated mustard ( Sinapis alba L.) seedlings is known to be controlled by phytochrome (Pfr) through a threshold mechanism. The Pfr threshold value required to suppress hypocotyl growth was low (3 times 10⁻²% Pfr, based on total phytochrome in the hypocotyl at 36 h after sowing = 100%). In the present study the question was addressed whether the threshold control by Pfr of hypocotyl elongation also operates in light-pretreated, partly de-etiolated seedlings after transfer to darkness. The experimental results show that this is the case. Calculation of the threshold level in far-red light pretreated seedlings led to a very low value (3 times 10⁻⁷%) compared to etiolated seedlings (3 times 10⁻²%). In red light pretreated seedlings the threshold level was calculated to be 9 times 10⁻⁷%. Since the light pretreatment affected the rate of degradation of phytochrome strongly (half-life of Ptot in continuous red light was found to be 35 min in far-red pretreated instead of 47 min in etiolated material), the difference in threshold level between far-red and red pretreated material cannot be interpreted unambiguously. However, the conclusion can be drawn that light nretreatment strongly increases the degradation rate of Pfr and decreases the threshold level.     

RAPID TRANSMISSION OF A PHYTOCHROME SIGNAL FROM HYPOCOTYL HOOK TO COTYLEDONS IN MUSTARD (Sinapis alba L.)

Abstract— In mustard ( Sinapis alba L.) cotyledons appearance of 'potential capacity' to phosphorylate is controlled by phytochrome (P_fr) through a threshold, all-or-none reaction. Phytochrome operates only if the hypocotyl hook is connected to the cotyledons for at least 2.5 min after the end of a 1 min inductive light pulse. Partial irradiations of hook and cotyledons show that in the case of induction of 'potential capacity' rapid transmission of the P_fr signal occurs from the hypocotyl hook to the cotyledons. Phytochrome within the cotyledons is totally ineffective in this regard. A 3 min red light pulse suffices for full transmission of the signal. It seems that an explanation of the facts we have observed requires a revision of the present views about communication between the organs of a plant.     

AN ANALYSIS OF PHYTOCHROME-MEDIATED THRESHOLD CONTROL OF HYPOCOTYL GROWTH IN MUSTARD (Sinapis alba L.) SEEDLINGS†

Hypocotyl elongation in mustard (Sinapis alba L.) seedlings is known to be controlled by phytochrome (P_fr) through a threshold response. This phytochrome-mediated threshold response was studied in detail with the following results: (i) The P_fr threshold value required to suppress hypocotyl growth is much lower (0.03% P_rr, based on total phytochrome in the hypocotyl at 36 h after sowing = 100%) than those threshold valued observed previously in threshold control by hook phytochrome of appearance of 'potential capacity for photophosphorylation' and lipoxygenase appearance in the mustard cotyledons (1.25% P_tr, based on total phytochrome in the hypocotyl at 36 h after sowing = 100%). This probably explains why hypocotyl elongation is so extremely sensitive to light, (ii) The P_fr threshold value controlling hypocotyl growth is a system constant, independent of total phytochrome content, developmental age and actual growth rate, (iii) Threshold control of hypocotyl elongation is unaffected by the removal of the cotyledons and half of the hook. However, removal of the whole hook totally eliminates any light control over the residual hypocotyl growth, (iv) After termination of the threshold control, the hypocotyl growth rate immediately returns to precisely that found in untreated dark control even though the partial growth rates of the different parts of the hypocotyl are quite different, relative to their dark controls. Obviously, the organ grows as an integrated unit.
It is concluded that the all-or-none threshold control over hypocotyl growth is exerted from the plumular hook. It appears that the hook can send off phytochrome all-or-none signals in both directions, to the cotyledons and to the hypocotyl.     

PHOTOTROPISM AND AXIS EXTENSION IN LIGHT-GROWN MUSTARD (Sinapis alba L.) SEEDLINGS‡

Abstract— The relationship between phototropism and axis extension was examined in light-grown mustard (Sinapis alba L.) seedlings using the low pressure sodium lamp (SOX)? technique to eliminate growth responses due to phytochrome. Addition of blue light caused no net inhibition of hypocotyl elongation, but plants showed a phototropic response. Curvature was caused by a simultaneous inhibition of growth on the illuminated side of the hypocotyl and an acceleration on the shaded side. Phototropism thus occurs independently of axis elongation and suggests that they are two separate processes. The results are inconsistent with the Blaauw theory of phototropism.     

PHYTOCHROME CONTROL OF DEGREENING OF ATTACHED COTYLEDONS and PRIMARY LEAVES OF MUSTARD (SINAPIS ALBA L.) SEEDLINGS*

Abstract— Mustard ( Sinapis alba L.) seedlings were grown on moistened vermiculite without added nutrients for 17 days in medium white light (4.6 lx). During this period cotyledons as well as primary leaves expressed signs of senescence as defined by the loss of chlorophyll(ide) a ('degreening'). While degreening was detectable 9 days after sowing in the cotyledons it was not measurable before 15 days after sowing in the primary leaves. Phytochrome retards the loss of chlorophyll(ide) a in both organs; however, it does not change the temporal pattern of greening and degreening.     

INVOLVEMENT OF PHYTOCHROME IN LIGHT CONTROL OF STEM ELONGATION IN CUCUMBER (Cucumis sativus L.) SEEDLINGS

A series of polycondensed aromatic N-heterocycles (acridine, benzo-f-quinoline 1,2,7,8-dibenzacridine and 3,4,5,6-dibenzacridine) were adsorbed from the gas phase and from liquid solution on highly dispersed silica gels with very different specific surface areas and pore sizes. The translational mobility of the adsorbed species was quantified by the triplet decay and the delayed fluorescence following bimolecular triplet-triplet annihilation after pulsed laser irradiation. The decay kinetics were analyzed with conventional second order rate equations and with the fractal approach. The first method is reliable without limitations on adsorbents with large pore diameters. It yields second order annihilation constants of 4 times 10¹²-6 times 10¹¹ dm² mol^?1 s^?1 depending on the masses and sterical requirements of the adsorbates. For the second method a spectral dimension d_s= 4/3 was used. This method is quantitatively applicable to all heterocycles adsorbed on silica gel 60 that have very small pore sizes. An activation energy of 4.9 ± 0.5 kcal mol^?1 was obtained for the translational diffusion of acridine on hydroxylated silica gel.     

IN VITRO SYNTHESIS OF PHYTOCHROME APOPROTEIN DIRECTED BY mRNA FROM LIGHT AND DARK GROWN AVENA SEEDLINGS*

Abstract— The aim of this work was to compare the translatability of poly-A-RNA from light- and dark-grown Arena seedlings, the product of which should be phytochrome. Polysomal poly-A-RNA was isolated from 4-day-old dark-grown Avena seedlings (Avena sativa L. cv. Garry) and translated in a rabbit reticulocyte lysate system. Immunoprecipitation by anti-phytochrome serum was used for measurements of specific phytochrome translation. Characterization of the translated and immunoprectpitated protein was performed by comparison with [₃₅S]-methionine in vivo-labelled phytochrome. The specificity of the precipitation was shown by parallel use of non-specific serum and by competitive inhibition of precipitation by exogenous unlabelled phytochrome isolated by affinity chromatography.     

BLUE LIGHT INDUCED PHOTOTRANSFORMATION OF PHYTOCHROME IN THE PRESENCE OF FLAVIN*

Abstract— Irradiation of the P_r form of phytochrome in the presence of flavin mononucleotide (FMN) which absorbs the actinic blue light yields P_fr at a rate greater than that in the absence of FMN. The actinic blue light absorbed by FMN enhances the phototransformation of P_r via the energy transfer from the former to the latter. On the other hand, the photoreversion of P_fr was inhibited by the presence of FMN when illuminated with blue light. The lack of photo-enhancement of the reversion of P_r, by blue light suggests that the P_fr chromophore (acceptor) transition dipole is virtually perpendicular to the FMN transition dipole, as the result of a chromophore reorientation in the P_r→P_fr phototransformation. The fact that blue light absorbed by flavin preferentially enhances the forward phototransformation of phytochrome while inhibiting the reversion may have an important implication in the high irradiance responses in plants in terms of a preferential accumulation of P_fr by blue light excitation.     

INTERACTION BETWEEN PHYTOCHROME AND THE BLUE LIGHT PHOTORECEPTOR SYSTEM IN Mougeotia*

Abstract— Face-to-profile chloroplast movement in Mougeotia was induced by sequences of strong blue and red short irradiations. This type of response occured only when blue light was applied prior to or simultaneously with red light, and far-red irradiation was necessary after the sequence to cancel the remaining gradient of the far-red absorbing form of phytochrome P_fr. The dependence of the response magnitude on blue and red light sequences was studied for a wide range of light durations and dark intervals. The relationship between the response and the dark interval points to the lack of direct coupling between phytochrome and blue-absorbing “cryptochrome”. It was postulated that a photoproduct having a life-time of2–3 min is formed by the blue-light-mediated reaction. This photoproduct interacts with phytochrome during its transformation or with its final P_fr form.     

CHANGES IN THE RATES OF PHOTOCONVERSION OF PHYTOCHROME DURING ETIOLATION IN MUSTARD SEEDLINGS

Abstract— The relative phytochrome photoconversion rates in cotyledons and hypocotylar hook of etiolating mustard ( Sinapis alba L.) seedlings were measured between 16 and 96 h after sowing. It was found that at constant fluence rates photoconversion rate in red light increases in both organs with time whereas the photoconversion rate in far-red (756 nm) light decreases with time of development. Since the isosbestic point remains constant, it was concluded that the observed changes cannot be attributed to changes of extinction coefficients. It was not possible, however, to decide whether the observed changes are due to changes of light attenuation or quantum yields.     

MODE OF COACTION OF PHYTOCHROME AND BLUE LIGHT PHOTORECEPTOR IN CONTROL OF HYPOCOTYL ELONGATION

Abstract The rate of hypocotyl longitudinal growth in seedlings of Sesamum indicum L. is strongly inhibited by continuous blue light (cBL)† and slightly by continuous far-red light while continuous red light (cRL) or red light pulses are hardly effective from 60 h after sowing onwards. Between 36 and 60 h after sowing the growth rate responds to red light pulses the effect of which is fully reversible by long wavelength far-red light. When seedlings are kept in cBL for 3 days and then treated with red light hypocotyl growth rate responds strongly. However, RL effectiveness decreases with time after transfer from BL to RL. BL → darkness transfer experiments with different levels of P_fr established at the beginning of darkness show that after a BL pretreatment phytochrome (P_fr) alone is capable of fully controlling growth rate. When white light (WL) is given no BL effect is detectable in weak WL. Only high light fluxes maintain a typical BL growth rate. At medium WL fluxes elongation rate returns gradually to the dark rate. The simplest explanation of the data is that light absorbed by a separate BL photoreceptor is necessary to maintain responsivity to P_fr. With increasing age of the seedlings the requirement for BL increases strongly. On the other hand, brief light pulses—given to demonstrate photoreversibility of phytochrome—remain equally effective provided that responsivity to P_fr exists.     

MODE OF COACTION BETWEEN UV-A AND LIGHT ABSORBED BY PHYTOCHROME IN CONTROL OF APPEARANCE OF RIBULOSE-1.5-BISPHOSPHATE CARBOXYLASE IN THE SHOOT OF MILO (Sorghum vulgare Pers.)

Abstract— In shoots of milo ( Sorghum vulgare Pers.) appearance of ribulosebisphosphate carboxylase (RuBPCase) and of translatable mRNA for its small subunit is stimulated strongly by red light (R, operating through phytochrome) and UV-A light (UV-A). Ultraviolet-A is more effective than R.
The mode of coaction between phytochrome and light absorbed by the blue/UV-A light photoreceptor ('cryptochrome') was analyzed in detail in case of enzyme appearance. Fluence rate dependencies, lagphases and the time course of the response are compatible with the view that UV-A intensifies a process which is occurring in R alone albeit at a lower rate.
With both light qualities the light effect is fully reversible by far-red light up to 1 h. This means that during this period only phytochrome (P_fr) controls the terminal response, i.e. the actual appearance of RuBPCase. During this 1 h period after the onset of light UV-A or R have no effect on the level of translatable mRNA for the small subunit of RuBPCase indicating that it requires more than 1 h for the light signal to affect gene expression.
When R and UV-A are given longer onset of escape from full reversibility is observed at the same time for both light qualities in the case of RuBPCase appearance. The extent of the reversible response is greater after UV-A pretreatment than after a R pretreatment.
It is argued that the data are consistent with the concept that phytochrome (P_fr) controls the terminal photoresponse, in the present case appearance of RuBPCase, while light absorbed via cryptochrome leads to an increase in responsiveness of the RuBPCase producing machinery towards P_fr.     

PHYTOCHROME INDUCTION OF PHOTOREACTIVATING ENZYME IN Phaseolus vulgaris L. SEEDLINGS*

Abstract— Photoreactivating enzyme (PRE) activity was measured in hypocotyls of Phaseolus vulgaris L. seedlings using a radioimmunoassay for thymine dimers. In dark-grown seedlings a five-fold increase in PRE activity was observed after 6 h of irradiation with blue or far-red light. Short time irradiations with red light were also effective. Reversibility of this red-light-effect by a subsequent short term irradiation with far-red light and also the high effectiveness of continuous far-red light indicate that PRE activity is under phytochrome control. This observation points to PRE induction via gene activation.     

THE EFFECT OF PROLONGED LIGHT EXPOSURE ON THE EFFECTIVENESS OF PHYTOCHROME IN ANTHOCYANIN SYNTHESIS IN TOMATO SEEDLINGS*

Abstract— The hypocotyl of the tomato ( Lycopersicon esculentum ) seedling synthesizes large amounts of anthocyanin if exposed to prolonged light. Single light pulses are totally ineffective. The involvement of phytochrome can be shown by light pulse treatments following a prolonged light exposure. It is predominantly the action of blue/UV light which leads to a high responsiveness of anthocyanin synthesis towards phytochrome. Moreover, the data suggest a phytochrome-independent action of blue/UV light, in particular of UV-B, on anthocyanin synthesis.     

CONTROL OF HYPOCOTYL GROWTH IN MUSTARD SEEDLINGS AFTER LIGHT-DARK TRANSITIONS

Abstract— Six hours of irradiation with white or far-red light strongly stimulates the 'end of day' inhibition of hypocotyl elongation in dark grown Sinapis alba L. Measurements of both reversion kinetics of the inductive light pulse and 'null point' experiments (Hillman, 1965. 1972) indicate that this response is controlled by stable phytochrome. The extent of the reversible response (i. e. length after far-red light minus length after red light) showed rhythmic oscillations after a 6, 48 and 54 h white light pretreatment. The rhythm is started by the transition from light to dark and neither phase nor amplitude is influenced by the level of P at this transition.     

THE BLUE ANOMALOUS EMISSION OF LARGE AND SMALL PHYTOCHROME*

–Small and immunoaffinity-purified large phytochrome (P_r and P_fr) show a so-called anomalous fluorescence with λ^em_max= 470 and 440 nm, respectively, when irradiated within the blue absorption band. Model studies indicate that this emission arises from a dipyrromethenone partial structure produced by a nucleophilic addition to the central carbon C-10 of the bilindione chromophore. The anomalous emitter of phytochrome is thus similar to one bilirubin conformer which has previously been found to contribute to the absorption and emission of the bile pigment.     

CONTROL BY PHYTOCHROME OF CHLOROPHYLL SYNTHESIS IN SEEDLINGS OF SORGHUM VULGARE

The strong effect of light pretreatments on the synthesis of chlorophyll-a and-b in the shoot of Sorghum vulgare (kept under saturating white light) can be attributed to phytochrome only. No specific blue light effect was found. The phytochrome system appears to function perfectly normally under these conditions. Escape from reversibility is not detectable up to approximately 40 min after the onset of an inductive red light pulse. Thereafter, escape is fast, being completed at approximately 2.5 h after the inductive light pulse.     

FLUENCE RATE COMPENSATION OF THE PERCEPTION OF RED: FAR-RED RATIO BY PHYTOCHROME IN LIGHT-GROWN SEEDLINGS*

Abstract— The hypothesis that phytochrome functions as a sensor of vegetational shade through the perception of the red: far-red photon fluence rate ratio requires that the mechanism of perception be compensated for wavelength-independent fluctuations in fluence rate (Smith, 1982). This paper seeks to establish the lower limit of fluence-rate compensation and to assess whether or not compensation is effective at the total fluence rates typical of herbaceous canopies. Using specially-designed cabinets, Sinapis alba L. (white mustard) seedlings were grown from germination under a range of total photosynthetically-active radiation (PAR = 400 to 700 nm) values and a range of red: far-red ratios. The data indicate that fluence-rate compensation is effective above a PAR value of ca. 60 μ.mol m² s'. Pretreating seedlings at high red: far-red ratio and a PAR level of 300 (μmol m₂S_-1for increasing periods of time led to an extension of fluence rate compensation to lower fluence rates. The results are discussed in relation to the photosynthetic competence of the seedlings grown under these conditions.     

THE MODE OF INTERACTION BETWEEN BLUE (UV) LIGHT PHOTORECEPTOR AND PHYTOCHROME IN ANTHOCYANIN FORMATION OF THE SORGHUM SEEDLING

Two non-photosynthetic photoreceptors (phytochrome and a blue light photoreceptor) are involved in light-mediated anthocyanin synthesis in the mesocotyl of Sorghum seedlings. The present study was undertaken to investigate the kind of interaction between phytochrome and the blue light photoreceptor. The data show that phytochrome (P_fr) can only act once a blue light effect has occurred. On the other hand, the blue light effect cannot express itself without P_fr. It is concluded that there is an obligatory dependency (or sequential interaction) between the blue light effect and the light effect occurring through phytochrome, although the blue light photoreaction per se is not affected by the presence or absence of phytochrome. The latter statement is based on the results of dichromatic experiments, i.e. simultaneous, high fluence rate irradiation with two kinds of light. Blue light can be replaced by UV light. It is not clarified yet whether the effect of blue and UV light is due to the same photoreceptor.