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.     

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.     

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.     

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.     

TEMPERATURE AND MYCOCHROME SYSTEM IN NEAR-UV LIGHT INDUCIBLE AND BLUE LIGHT REVERSIBLE PHOTOINDUCTION OF CONIDIATION IN Alternaria tomato*

Abstract— When dark-grown colonies were exposed to near-UV light, conidiophore formation was induced, and conidia developed during a subsequent dark period. Simultaneous exposure to near-UV and blue light inhibited induction of conidiation. The inductive effect of near-UV light irradiation was greatly enhanced by treating colonies with low temperatures for 4 h during the 2nd-6th hour of incubation in the dark following inductive irradiation: the enhancement was greatest at 21^°C. The extent of inhibition by blue light increased with the temperature between 10 and 28^°C. This diminution by low temperature was greatest when colonies previously kept at a low temperature were exposed to inductive irradiation: the longer the duration between inductive irradiation and temperature treatment, the lower the diminution. Higher fluences of blue light were required to suppress conidial induction at lower temperature. Thus, it is evident that the inductive effect of near-UV light irradiation on conidiation and the suppressive effect of blue light irradiation are each responsive to different temperatures applied at different times.     

SYNERGISTIC INTERACTION BETWEEN THE TRYPTOPHAN PHOTOPRODUCTS AND NEAR-ULTRAVIOLET LIGHT IN AMOEBA

Abstract. Both S- and G₂-phase cells of amoeba are found to be sensitized by coirradiation of cells with Tryptophan (Trp). At a fluence of 1.5 times 10⁵ Jm^-2 to which G₂ phase cells are resistant, they show a drastic (8-fold) increase in sensitivity when irradiated in 0.5% Trp solution. Freshly irradiated Trp solution is found to be lethal to G₂ cells that have received a sublethal fluence of near-UV light. Irradiated Trp, however, does not kill unirradiated amoeba.
Nuclear transplantation experiments have shown that the lethality of cells when coirradiated with Trp is due to damage located predominantly in the cytoplasm. It is suggested that sublethal fluence of near-UV light makes the cells "leaky" to the toxic photooxidation products of Trp solution.     

RELATIVE IMPORTANCE OF BLUE LIGHT AND LIGHT ABSORBED BY PHYTOCHROME IN GROWTH OF MUSTARD (Sinapis alba L.) SEEDLINGS*

Abstract— Hypocotyl straight growth in mustard (Sinapis alba L.) responds very strongly and in precisely the same way to low fluence rate red (RL) and white light (WL). The effect of weak light can be attributed fully to light absorption by phytochrome. Only with increasing fluence rate an effect of blue light (BL) comes into play which cannot be explained by the action of phytochrome. However, this specific action of BL can be demonstrated in hypocotyl growth of mustard seedlings only up to 5 days after sowing (25°C). With older seedlings control of hypocotyl growth seems to be exerted exclusively via phytochrome. Regarding the far-red light dependent “high irradiance reaction” (FR-HIR) it was found that it plays a dominant role in growth of mustard only during a relatively short period. It tends to disappear in favor of a RL-HIR between 3 and 4 days after sowing. It is concluded that the seedling exhibits a largely endogenous temporal pattern of responsiveness to light. Phototropism of the mustard seedling can be elicited by low fluence rates (< 1 mW m^?2) of unilateral BL. This same light has no effect on straight growth. It is concluded that BL-dependent phototropic growth response of a hypocotyl and the effect of BL on longitudinal growth of the hypocotyl are unrelated phenomena.     

PHOTOPHOBIC BEHAVIORAL RESPONSES OF EUGLENA IN A LIGHT INTENSITY GRADIENT AND THE KINETICS OF PHOTORECEPTOR PIGMENT INTERCONVERSIONS*

Abstract— Accumulation of Euglena gracilis in small illuminated regions called light traps is due to a phobic response to the diminished light intensity at the boundary of the region. The rate of such accumulation of cells was measured as functions of both the light intensity within the trap and the change of intensity at the boundary of the trap. The initial rate of accumulation of a population of cells was taken to be a direct measure of the phobic response of a single typical cell. The data indicate that the strength of the behavioral response in a single cell may be described as being proportional, to the rate of change of the amount of photochemically active form of a photoreceptor pigment molecule which can exist in two predominant forms.     

EFFECT OF UVA AND BLUE LIGHT ON PORPHYRIN BIOSYNTHESIS IN EPIDERMAL CELLS*

Abstract— To study porphyrin biosynthesis in normal human keratinocytes and A431 cells derived from human epidermoid carcinoma, cultured cells were incubated with delta-aminolevulinic acid (ALA), the precursor of porphyrin synthesis, and accumulation of porphyrins was measured spectrofluorometrically. Both human keratinocytes and A431 cells accumulated porphyrins in a time-dependent and a dose-dependent fashion. Protoporphyrin was the predominant porphyrin accumulated by both cell types. Porphyrin accumulation was enhanced by Ca Mg ethylene-diaminetetraacetic acid, a ferrochelatase inhibitor, and the enhancement was reversed by the addition of iron, suggesting the utilization of iron by ferrochelatase. The effect of light on porphyrin accumulation was evaluated by exposing the ALA-loaded A431 cells to ultraviolet-A (UVA) and blue light radiation, followed by continued incubation with ALA for 2–48 h. There was an enhancement of porphyrin accumulation 2–48 h after the radiation as compared with nonirradiated controls. Consistent with this finding, ferrochelatase activity decreased in these cells at 24 h and 48 h. These data demonstrate that human keratinocytes and A431 cells are capable of porphyrin biosynthesis, and that exposure of porphyrin-containing A431 cells to light, which includes the Soret band spectrum, decreases the ferrochelatase activity, which is responsible, at least in part, for the further increase in porphyrin level.     

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.     

LIGHT PERCEPTION BY PHYTOCHROME in vivo*

The rate of phytochrome photoconversion in cotyledons and hypocotyls of dark-grown seedlings of Cucurbita pepo varies significantly depending on the characteristics of the light field and the position of the sample in the light field. The overall pattern of the effects of light reflectance conditions and angle of incidence of light on phytochrome photoconversion in vivo is predictable and consistent with the results of physiological studies; however, the extent of the effects is much more difficult to predict with a satisfactory degree of approximation. The differences between phytochrome photoconversion kinetics measured in vivo and those predicted from photochemical parameters of purified phytochrome are rather large and suggest that the use of predictions as estimates of the state of phytochrome in vivo may be subject to significant limitations.     

PHOTOCONVERSION OF PHYTOCHROME IN VIVO STUDIED BY DOUBLE FLASH IRRADIATION IN Mougeotia AND Avena

Abstract— The kinetics of phytochrome phototransformation from the red-absorbing form (P_r) to the far-red-absorbing form (P_fr) in vivo at 22°C were studied using a double flash apparatus with 1-ms flashes. Photoconversion by simultaneous flashes of red light saturates at a low P_fr level, indicating the possible attainment of a photoequilibrium between the excitation of P_r and the photoreversion of intermediates in the course of the I-ms flashes. At saturation energy, simultaneous flashes resulted in about 50% as much P_fr as was produced by saturating irradiation with 5 s red light. Intermediates of the phototransformation pathway were analysed by separating two red or a red and a far-red flash by variable dark intervals. In both plants phototransformation intermediates with half-lives < 1 ms occur, but they are too short-lived to characterize by our method. The subsequent intermediates have half-lives of about 7 ms and 150 ms in A vena , 2 ms and 10 ms in Mougeotia. The conversion from P_r to P_fr seems to be completed 1 s after the red flash in Avena. In the alga Mougeotia , P_fr formation seems to be finished within only 50 ms after the inducing red flash. The kinetics obtained from physiological and spectrophotometric experiments with Avena mesocotyls are almost identical. These observations indicate that the physiological response corresponds directly to the amount of P_fr produced and not to phototransformation intermediates or "cycling" between P_r and P_fr.     

CORRELATION BETWEEN ABSORBANCE CHANGES AND A PHYSIOLOGICAL RESPONSE INDUCED BY BLUE LIGHT IN NEUROSPORA

Abstract— Absorbance changes due to the photoreduction of a b -type cytochrome are associated with many biological blue light-controlled processes. Evidence is presented for their causal relationship with the perception of light which induced conidia formation under conditions of starvation in Neurospora crassa mutant albino band, but not with light-induced phase shifts of conidiation bands.     

PHOTORECEPTOR INTERACTION IN PLANT PHOTOMORPHOGENESIS: THE LIMITS OF EXPERIMENTAL TECHNIQUES AND THEIR INTERPRETATIONS

Abstract— Problems concerning the interpretation of interactions of higher plant photomorphogenetic receptors are discussed. The theory that action of a blue light photoreceptor serves only to maintain responsiveness to phytochrome (Responsiveness Theory) is demonstrated to be unable to be properly tested with present techniques. This theory is also unable to explain experimental results any better than an alternative theory that a blue light photoreceptor may require the presence of the active form of phytochrome to express its activity (Presence Theory). This tatter theory is also incapable of being fully tested. There does not appear to be an adequate current theory to explain photoreceptor interactions. Other issues discussed include the use of displacement transducers in growth studies, the induction of phytochrome-type responses by blue light, and the relative importance of the photoreceptors. New data are introduced on the effect of blue light in the end-of-day growth response to phytochrome of the light-grown Cucumis sativus L. hypocotyl, and on the light equivalence principle in the same species.     

THE BLUE LIGHT RESPONSE OF STOMATA: PULSE KINETICS AND SOME MECHANISTIC IMPLICATIONS*

Abstract— Intact leaves of Commelina communis irradiated with high fluence rates of red light, showed discrete increases in stomatal conductance in response to pulses (1-100 s) of blue light (250 μmol m^?2 s^?1). Red light pulses were ineffective, indicating that the conductance increases were not mediated by photosynthesis and that they constitute a specific stomatal response to blue light. The response peaked 15 min after the pulse and was completed within50–60 min. Conductance increases were proportional to pulse duration up to about 30 s and saturated at longer exposures. The relationship between stomatal responses and pulse duration approximately fitted an exponential function, with a t 9s. Pulse responses at two different fluences indicated that reciprocity held. Responses to two consecutive pulses varied with time between pulses. A saturating pulse applied immediately after a preceding one induced no additional response; two saturating pulses 50 min apart caused two identical, consecutive responses. Total increases in conductance induced by two pulses separated by intermediate time intervals increased with time between pulses with a = 9 min. These results point to a blue light-dependent photoconversion of a molecular form, with the activity of the photoconversion product decaying in a thermal reaction. Under continuous blue light, prevailing fluence rates and rates of the light and thermal reactions are postulated to determine steady-state activities of the photoconversion product and proportional increases in conductance levels. These findings have implications for the environmental and metabolic roles of the stomatal response to blue light.     

BLUE AND ULTRAVIOLET-B LIGHT PHOTORECEPTORS IN PARSLEY CELLS

Abstract— Ultraviolet-B (UV-B) and blue light photoreceptors have been shown to regulate chalcone synthase and flavonoid synthesis in parsley cell cultures. These photoreceptors have not yet been identified. In the present work, we studied UV-B photoreception with physiological experiments involving temperature shifts and examined the possible role of flavin in blue and UV-B light photoreception. Cells irradiated with UV-B light (0.5–15 min) at 2°C have the same fluence requirement for chalcone synthase and flavonoid induction as controls irradiated at 25°C. This is indicative of a purely photochemical reaction. Cells fed with riboflavin and irradiated with 6 h of UV-containing white light synthesize higher levels of chalcone synthase and flavonoid than unfed controls. This effect did not occur with blue light. These results indicate that flavin-sensitization requires excitation of flavin and the UV-B light photoreceptor. The in vivo kinetics of flavin uptake and bleaching indicate that the added flavin may act at the surface of the plasma membrane. In view of the likely role of membrane-associated flavin in photoreception, we measured in vitro flavin binding to microsomal membranes. At least one microsomal flavin binding site was solubilized by resuspension of a microsomal pellet in buffer with high KPi and NaCl concentrations and centrifugation at 38000 g. The 38000 g insoluble fraction had much greater flavin binding and contained a receptor with an apparent K_D of about 3.6 μM and an estimated in vivo concentration of at least 6.7 × 10–⁸M. Flavin mononucleotide, roseoflavin, and flavin adenine dinucleotide can compete with riboflavin for this binding site(s), although each has lower affinity than riboflavin. Most microsomal protein was solubilized by resuspension of the microsomal pellet in non-denaturing detergents and centrifugation at 38 000 g ; however, this inhibited flavin binding, presumably because of disruption of the environment of the flavin receptor. The parsley microsomal flavin binding receptor(s) have a possible role in physiological photoreception.     

BLUE AND GREEN LIGHT EFFECTS,AND THEIR PHOTOREVERSIBILITY,UPON ZOOSPORE PRODUCTION AND THE SYNCHRONIZED CYCLE IN PROTOSIPHON BOTRYOIDES KLEBS*

Abstract —The synchronized asexual cycle in Protosiphon botryoides Klebs is described. A blue light (430 nm) inhibition of zoospore production can be photoreversed by green light (550 nm). Similarly, a green light (550 nm) stimulation of zoospore production can be photoreversed by blue (430 nm) light. 430 nm light also shortens the period of zoospore motility, and 550 nm light lengthens this period.     

INTERACTION BETWEEN UPOSOMES AND MONOLAYERS IN THE PRESENCE OF CALCIUM

Abstract

The interaction between phospholipid vesicles (phosphatidylcholine : phosphatide acid, 90:10 w/w) and phosphatidylcholine : cholesterol (70:30, molar ratio) monolayers at air/water interfscks has been studied at. several concentrations of calcium cation ( Ca²⁺). The liposome vesicles were SUVs and MLVS.

The vesicles interact with the monolayers, rapidly causing a large increase in surface pressure. Limiting values of surface pressure, 2.07-6.99 mN.m-1 for SUVs, and 7.01-11.11 mN.m^?1 for MLVs, were reached in less than 40?min.

Calcium ion concentration affects the liposome size in MLVs, producing an increase of gyration radius. The SUVs are little influenced. The change in size can be due to a variation of liposome composition induced by calcium: cholesterol molecules can migrate from monolayer to liposomes and the redistribution of exchanged lipids in the outer bilayer can also explain the size variation.