Control of chlorophyll a synthesis by phytochrome and cryptochrome in the red alga Corallina elongata Ellis et Soland

Chlorophyll a synthesis in the red alga Corallina elongata is controlled by phytochrome and by a specific blue light photoreceptor. Although the estimated photoequilibrium of phytochrome is similar in blue and red light, the amount of chlorophyll accumulated is greater in blue light, which implies the action of cryptochrome, according to the criteria for the specific blue light photoreceptor involvement. The amount of chlorophyll synthesized is greater when the level of photoequilibrium approaches 65% (in blue and red light) than with higher levels (72.7% in white light and 70.8% in green light). The action of phytochrome is demonstrated by the induction of chlorophyll synthesis after red pulses and the reversion after far red pulses. The reversion is not complete but the percentage of reversibility is high (85-90%). The amount of chlorophyll accumulated is greater in darkness after the application of red light pulses than in white light after the same light pulses. The induction of chlorophyll synthesis is greater after red pulses than after continuous red light. The existence of a fast destruction of chlorophyll in continuous light is observed. This destruction is greater in the high photoequilibrium of phytochrome (70-72%). The turnover times and the induction mechanism of chlorophyll synthesis must be very fast. This indicates the existence of a possible rapid adaptation to the change in light quality and intensity in the marine system.     

Involvement of phytochrome and a blue light photoreceptor in UV-B induced flavonoid synthesis in parsley (Petroselinum hortense Hoffm.) cell suspension cultures

Flavonoid synthesis in cell suspension cultures of parsley (Petroselinum hortense Hoffm.) occurs only after irradiation with ultraviolet light (UV), mainly from the UV-B (280–320 nm) spectral range. However, it is also controlled by phytochrome. A P_fr/P_tot ratio of approximately 20% is sufficient for a maximum phytochrome response as induced by pulse irradiation. Continuous red and far red light, as well as blue light, given after UV, are more effective than pulse irradiations. The response to blue light is considerably greater than that to red and far red light. Continuous red and blue light treatments can be substituted for by multiple pulses and can thus probably be ascribed to a multible induction effect. Continuous irradiations with red, far red and blue light also increase the UV-induced flavonoid synthesis if given before UV. The data indicate that besides phytochrome a separate blue light photoreceptor is involved in the regulation of the UV-induced flavonoid synthesis. This blue light receptor seems to require the presence of P_fr in order to be fully effective.Abbreviations HIR high irradiance response - P_fr far red absorhing form of phytochrome - P_tet total phytochrome - UV ultraviolet light     

Photoregulation of Protochlorophyllide Oxidoreductase and Rubisco Large Subunit Accumulation in Phytochrome A-Deficient Transgenic Tobacco Plants

Some morphogenetic responses, induced by far red (FR) light in tobacco plants (Nicotiana tabacum L.), were studied. The inhibitory effect of FR irradiation on chlorophyll synthesis in transgenic plants with reduced phytochrome A content was almost absent. Phytochrome A-mediated repression of the por gene was demonstrated with the use of polyclonal antiserum against protochlorophyllide oxidoreductase. Continuous FR light induced the accumulation of Rubisco large subunits in wild-type but not in transgenic tobacco plants. Our data confirm the suggestion that phytochrome A mediates photoregulation of the synthesis of these proteins.     

Action of red and far red light on ribosome formation in cabbage seedlings

The action of light on ribosome formation was examined in the cabbage seedlings, a system extensively used in the studies of anthocyanin synthesis. Ribosomes were extracted 18 h after the beginning of the irradiation and separated by sucrose gradient centrifugation. In the cotyledons of dark-grown cabbage seedlings, a brief red light induces an increase both in total ribosomes and in the fraction present as polysomes; the effect of red light is reversed by far red light, indicating the involvement of phytochrome in polysome formation in cabbage seedlings. Continuous red and continuous far red light are about equally effective in bringing about an increase of total ribosomes and of the polysome fraction. Streptomycin, which inhibits chlorophyll synthesis and chloroplast development, and enhances anthocyanin synthesis in cabbage seedlings, causes a decrease of total ribosomes and of the fraction present as polysomes. In hypocotyls, the red-far red reversibility is evident only for the polysome content and streptomycin does not decrease the polysome/monosomo ratio as it does in cotyledons.     

Photocontrol of the synthesis of chlorophyll a and phycocyanin in the cyanobacterium Calothrix crustacea Schousboe]

Chlorophyll a and phycocyanin synthesis in the cyanobacterium Calothrix crustacea Schousboe (ecophene Rivularia bullata) have been studied in white light after the application of red and green light pulses. The light quality produces a complementary pattern in the pigment synthesis. Chlorophyll synthesis is stimulated by red light pulses whereas phycocyanin synthesis is by green light pulses. Because the effect of red light on chlorophyll synthesis shows some far-red photoreversibility, the action of phytochrome is proposed. The green light effect on phycocyanin synthesis is only partially reversed by far-red light. This reversion is lost after incubation in white light for two hours. The effect of green light on phycocyanin synthesis could not only be due to phytochrome since theoretically in green light the level of the active form of phytochrome is lower than in red light. Thus, the action of a specific green light photoreceptor is proposed.     

Phytochrome-mediated induction of phenylalanine ammonia-lyase in the cotyledons of tomato (Lycopersicon esculentum Mill.) plants

Phenylalanine ammonia-lyase (PAL; EC 4.3.1.5.) induction in cotyledons from 96-h dark-grown Lycopersicon esculentum Mill. was studied in response to continuous light and hourly light pulses (blue, red, far red). The increases of PAL promoted by blue and red pulses are reversed completely by immediately following 758 nm irradiations. The response to continuous red light could be substituted for by hourly 6-min red light pulses. The effect of continuous red treatments is mainly due to a multiple induction effect of phytochrome. In contrast to red light, hourly light pulses with far red and blue, light can only partially substitute for continuous irradiation. The continuous blue response could be due to a combination of a multiple induction response and of a high irradiance response of phytochrome. The continuous far red response, could represent a high irradiance response of phytochrome. Dichromatic irradiations indicate that phytochrome is the photoreceptor controlling the light response (PAL) in tomato seedlings.Abbreviations Norflurazon NF-4-chloro-5-(methylamino)-2-(,,,-trifluoro-m-tolyl)-3 (2H) pyridazinone - PAL phenylalanine ammonia-lyase - phytochrome photoequilibrium P_fr/P_tot - P_fr far-red absorbing form of phytochrome - P_r red absorbing form of phytochrome - P_tot total phytochrome: P_r+P_fr     

The involvement of phytochrome in controlling chlorophyll and 5-aminolevulinate formation in a gymnosperm seedling (Pinus sylvestris)

Chlorophyll a (Chl a) accumulation in the cotyledons of Scots pine seedlings (Pinus sylvestris L.) is much higher in the light than in darkness where it ceases 6 days after germination. When these darkgrown seedlings are treated with continuous white light (3,500 lx) a 3 h lag phase appears before Chl a accumulation is resumed. The lag phase can be eliminated by pretreating the seedlings with 7 h of weak red light (0.14 Wm^-2) or with 14 red light pulses separated by relatively short dark periods (<100 min). The effect of 15s red light pulses can be fully reversed by 1 min far-red light pulses. This reversibility is lost within 2 min. In addition, the amount of Chl a formed within 27 h of continuous red light is considerably reduced by the simultaneous application of far-red (RG 9) light. It is concluded that phytochrome (P_fr) is required not only for the elimination of the lagphase but also to maintain a high rate of Chl a accumulation in continuous light. Since accumulation of 5-aminolevulinate (ALA) responds in the same manner as Chl a accumulation to a red light pretreatment it is further concluded that ALA formation is the point where phytochrome regulates Chl biosynthesis in continuous light. No correlation has been found between ALA and Chl a formation in darkness. This indicates that in a darkgrown pine seedling ALA formation is not rate limiting for Chl a accumulation.Abbreviations Chl chlorophyll(ide) - PChl protochlorophyll(ide) - ALA 5-aminolevulinate - P_r the red absorbing form of phytochrome - P_fr the far-red absorbing form of phytochrome - P_tot total phytochrome ([P_r]+[P_fr])     

Chloroplast ribosomal ribonucleic acid synthesis in cultured spinach leaf tissue

Chloroplast rRNA synthesis was studied in spinach leaf tissue cultured under sterile conditions which eliminate bacterial rRNA synthesis. The synthesis was inhibited by darkness, but concomitant cytoplasmic rRNA synthesis was unaffected. A complex pattern of labelled rRNA precursors was found in extracts from cultured leaf tissue by using polyacrylamide-gel electrophoresis. However, differences between the precursor profiles of leaf tissue cultured in the light and in the dark could not be correlated with chloroplast rRNA synthesis since large amounts of high-molecular-weight precursors of cytoplasmic rRNA dominated the pattern in both cases. A double-isotope-labelling technique was used, which enabled light-stimulated rRNA synthesis to be studied in whole leaf tissue. Two rapidly labelled RNA species of molecular weights 1.15x10(6) and 0.65x10(6) were detected, which were thought to have possible precursor significance in the synthesis of mature chloroplast rRNA of molecular weights 1.04x10(6) and 0.56x10(6) respectively. Cycloheximide treatment resulted in the accumulation of RNA of molecular weight 1.8x10(6), whose function is unknown.     

The participation of phytochrome in the signal transduction pathway of salt stress responses in Mesembryanthemum crystallinum L.

Continuous irradiation of Mesembryanthemum crystallinum plantswith light of equal amounts of photosynthetically active radiation,but widely different red:far red ratios was used to intervenein phytochrome-mediated signal transduction pathways in thepresence and absence of salt stress. Light with a low ratioof red:far red (in contrast to light with a high ratio of red:farred), caused induction of PEP carboxylase activity, accumulationof the CAM isoform of PEP carboxylase, and the accumulationof malate anion. Taking these as indicators of CAM inductionit is concluded that phytochrome can participate in the signaltransduction pathway leading to CAM in M. crystallinum. A lowratio of red: far red light acted synergystically with saltstress in the induction of these CAM indicators. The simplestinterpretation of this interaction is that the phytochrome-mediatedeffects and salt stress effects acted on the same signal transductionpathway. The accumulation of pinitol was also increased by light witha low ratio of red:far red, consistent with the existence ofa stress syndrome in M. crystallinum which utilizes a commontransduction pathway. A low ratio of red:far red light induced a strong shade avoidanceresponse and, compared to light with a high red:far red ratio,modified chlorophyll content and betacyanin pigment complement. Plants grown in light with a low ratio of red:far red floweredearlier than plants grown in light with a high red:far red ratio. It is concluded that phytochrome can participate in the signaltransduction pathway leading to the induction of both CAM andthe processes which result in pinitol accumulation and pigmentationin M. crystallinum, as well as in the mediation of shade avoidanceand flowering responses. Key words: Mesembryanthemum crystallinum, CAM, phytochrome, signal transduction, drought stress     

Red-light and blue-light photoreceptors controlling chlorophyll a synthesis in the red alga Porphyra umbilicalis and in the green alga Ulva rigida

Chlorophyll synthesis is stimulated by red light in the green alga Ulva rigida C. Ag. and in the red alga Porphyra umbilicalis (L.) Kützing. Because the effect of red light showed some far-red reversibility in successive red and far-red light treatments, the involvement of phytochrome or a phytochrome-like photoreceptor is suggested. The extent of the response is dependent on exposure and photon fluence rate of red-light pulses. In addition to the effect of red light, a strong stimulation of chlorophyll synthesis by blue light was only observed in Ulva rigida. The effect of blue light shows also some far-red reversibility. In the green alga the accumulated chlorophyll is higher after blue light pulses than after red light pulses. In Porphyra umbilicalis , however, the contrary is observed. In Ulva rigida the involvement of a blue light photoreceptor in addition to phytochrome or a phytochrome-like photoreceptor is proposed. The different responses to red and blue light in both algae are explained in terms of their adaptation to the natural light environment.     

Photocontrol of Chlorophyll Loss in Papaya Leaf Discs

Both red and blue light pulses are separately shown to retarddark-stimulated chlorophyll loss of papaya leaf discs suggestingparticipation of phytochrome and blue light photoreceptors inregulating the pigment loss. The red light effect is fully reversibleby far-red light. The partial failure of far-red pulses to reversethe action of blue light suggests that blue light effect maynot be entirely through the phytochrome action. The apparentineffectiveness of continuous white light to check the chlorophyllloss is attributed to a balance of photooxidation and photoprotectionof the pigment. The interaction of blue light and kinetin at its different concentrationssuggests that the effect of interactions is additive. The bluelight effect in retarding chlorophyll loss is partly independentof the hormone level. (Received December 10, 1985; Accepted August 25, 1986)     

Events Surrounding the Early Development of Euglena Chloroplasts: I. Induction by Preillumination 1

Preillumination, followed by a dark period prior to exposure of dark-grown nondividing cells of Euglena gracilis var. bacillaris to normal lighting conditions for chloroplast development, results in potentiation, or abolishment of the usual lag in chlorophyll accumulation. The degree of potentiation is a function of the length of the preillumination period, the intensity of preilluminating light, and the length of the dark period interposed before re-exposure to continuous light for development. The optimal conditions are found to be: 90 minutes of preillumination with white light at an intensity greater than 30 microwatts per square centimeter (14 foot candles) followed by a dark period of at least 12 hours. Reciprocity is not found between duration and intensity of preilluminating light. Preillumination with blue light and red light was found to be the most effective in promoting potentiation, and the ratio of effectiveness of blue to green to red is consistent with protochlorophyll-(ide) being the photoreceptor. Although red light is effective, there is no reversal by far red light, and these facts, taken together with the effectiveness of blue light, suggest that the phytochrome system is not involved. The amount of chlorophyll formed at the end of preillumination is proportional to the resulting potentiation, suggesting that the amount of protochlorophyll(ide) removed or chlorophyll(ide) formed regulates this phenomenon. Potentiated and nonpotentiated cells show comparable rates of protochlorophyll(ide) resynthesis, suggesting that this is not the limiting factor in nonpotentiated cells. Although light is required for protochlorophyll(ide) conversion in chlorophyll synthesis, a brief preillumination seems also to initiate the production of components in the subsequent dark period which, in nonpotentiated cells, are ordinarily synthesized during the lag period under continuous illumination. These components are necessary to sustain maximal rates of subsequent chlorophyll accumulation.     

Control of chloroplast formation by light.

Light controls the formation of plastid ultrastructure and the synthesis of chlorophyll, plastid membrane constituents and Calvin cycle enzymes. A respective light-mediated regulation of the genetic apparatus in the nucleus and the plastid compartment has been reported. Three photoreactions are involved in the regulation: (1) the protochlorophyll (ide) leads to chlorophyll (ide) a photoconversion, (2) the formation of physiologically active phytochrome and (3) light absorption by a blue light receptor (cryptochrome). The chloroplast formation in higer plants is chiefly controlled by active phytochrome, while in lower plants cryptochrome is the prevailing regulatory factor.     

Inhibition of carotenoid biosynthesis by the herbicide SAN 9789 and its consequences for the action of phytochrome on plastogenesis

Treatment of the mustard (Sinapis alba L.) seedling with the herbicide SAN 9789 inhibits synthesis of colored carotenoids and interferes with the formation of plastid membrane lipids without affecting growth and morphogenesis significantly. In farred light, which is hardly absorbed by chlorophyll, development of plastid ultrastructure, synthesis of ribulosebisphosphate carboxylase and synthesis of chlorophyll are not affected by SAN 9789. It is concluded that normal phytochrome actions on plastid structural development, protein and chlorophyll syntheses are not affected by the absence of carotenoids provided that there is no significant light absorption in chlorophyll. The findings show that the inhibition of synthesis of one set of plastid membrane components (the carotenoids) does not stop synthesis of other components such as chlorophyll and does not halt membrane assembly. Supplementary experiments with the closely related compound SAN 9785, which affects the amount and composition of plastid lipids but not carotenoid and chlorophyll syntheses, suggest that the effect of the herbicide SAN 9789 is due exclusively to its inhibition of synthesis of colored carotenoids. In the presence of SAN 9789 white or red light at high fluence rate causes photodestruction of chlorophyll and ribulosebisphosphate carboxylase and photodecomposition of thylakoids. These effects are interpreted as resulting exclusively from the self-photooxidation and photosensitizing action of chlorophyll once the protection by carotenoids of chlorophyll against self- and sensitized photooxidation is lost.Abbreviations Carboxylase ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) - Chl chlorophyll a plus chlorophyll b - PAL phenylalanine ammonia-lyase (EC 4.3.1.5) - SAN 9789 -chloro-5-(methylamino)-2-(, , -trifluoro-m-tolyl)-3 (2H) pyridazinone - SAN 9785 4-chloro-5-(dimethylamino)-2-phenyl-3(2H)-pyridazione. SAN 9789 is sold commercially under the trade name Norflurazon - fr far red - wl white light     

Altered Phytochrome Regulation of Greening in an aurea Mutant of Tomato

A brief pulse of red light accelerates chlorophyll accumulation upon subsequent transfer of dark-grown tomato (Lycopersicon esculentum) seedlings to continuous white light. Such potentiation of greening was compared in wild type and an aurea mutant W616. This mutant has been the subject of recent studies of phytochrome phototransduction; its dark-grown seedlings are deficient in phytochrome, and light-grown plants have yellow-green leaves. The rate of greening was slower in the mutant, but the extent (relative to the dark control) of potentiation by the red pulse was similar to that in the wild type. In the wild type, the fluence-response curve for potentiation of greening indicates substantial components in the VLF (very low fluence) and LF (low fluence) ranges. Far-red light could only partially reverse the effect of red. In the aurea mutant, only red light in the LF range was effective, and the effect of red was completely reversed by far-red light. When grown in total darkness, aurea seedlings are also deficient in photoconvertible PChl(ide). Upon transfer to white light, the aurea mutant was defective in both the abundance and light regulation of the light-harvesting chlorophyll a/b binding polypeptide(s) [LHC(II)]. The results are consistent with the VLF response in greening being mediated by phytochrome. Furthermore, the data support the hypothesis that light modulates LHC(II) levels through its control of the synthesis of both chlorophyll and its LHC(II) apoproteins. Some, but not all, aspects of the aurea phenotype can be accounted for by the deficiency in photoreception by phytochrome.     

Induction of delta-Aminolevulinic Acid Formation in Etiolated Maize Leaves Controlled by Two Light Systems

A short illumination of etiolated maize (Zea mays) leaves with red light causes a protochlorophyll(ide)-chlorophyll(ide) conversion and induces the synthesis of δ-aminolevulinic acid (ALA) during a subsequent dark period. In leaves treated with levulinic acid, more ALA is formed in the dark than in control leaves. Far red light does not cause a conversion of protochlorophyll(ide) into chlorophyll(ide) and does not induce accumulation of ALA in the dark. Both red and far red preilluminations cause a significant potentiation of ALA synthesis during a period of white light subsequent to the dark period. The results indicate a dual light control of ALA formation. The possible role of phytochrome and protochlorophyllide as photoreceptors in this control system is discussed.     

Circadian Rhythmicity in Excised Samanea Pulvini: II. Resetting the Clock by Phytochrome Conversion

Excised Samanea saman pulvini were incubated in H(2)O or 50 mm sucrose in darkness for 100 to 152 hours except for brief exposures to red or far red light, and angles of opening measured periodically. When pulvini are incubated in H(2)O, the rhythm damps in the open position after two to three cycles irrespective of the light treatments, but when sucrose is available, the now persistent oscillations show large red, far red-regulated effects on phase, amplitude, mesor slope, and entrainment. Single red light pulses rephase the rhythm, with a phase response curve that resembles that reported for other plants and animals; such rephasing is prevented by immediately subsequent far red light, indicating that phytochrome is the photoreceptor. Red light pulses repeated every 24 hours entrain the rhythm, and also prevent damping if presented at an appropriate part of the cycle.     

Light requirement,phytochrome and photoperiodic induction of flowering of Pharbitis nil Chois

For dark-grown seedlings of Pharbitis nil capacity to flower in response to a single inductive dark period was established by 24 h white, far-red (FR) or ruby-red (BCJ) light and by a skeleton photoperiod of 10 min red (R)-24 h dark-10 min R. FR alone was ineffective without a brief terminal (R) irradiation, confirming that the form of phytochrome immediately prior to darkness is a crucial factor for flowering in Pharbitis. The magnitude of the flowering response was significantly greater after 24 h FR or white light (WL) (at 18° C and 27° C) than after two brief skeleton R irradiations, but the increased flowering response was not attributable to photosynthetic CO₂ uptake because this could not be detected in seedlings exposed to 24 h WL at 18° C. Photophosphorylation could have contributed to the increased flowering response as photosystem I fluorescence was detectable in plants exposed to FR, BCJ, or WL, but there were large differences between flowering response and photosystem I capacity as indicated by fluorescence. We conclude that phytochrome plays a major role in photoresponses regulating flowering. There was no simple correlation between developmental changes, such as cotyledon expansion and chlorophyll formation during the 24-h irradiation period, and the capacity to flower in response to a following inductive dark period. Changes in plastid ultrastructure were considerable in light from fluorescent lamps and there was complete breakdown of the prolamellar body with or without lamellar stacking at 27 or 18° C, respectively, but plastid reorganization was minimal in FR-irradiated seedlings.Abbreviations BCJ irradiation from photographic ruby-red lamps - FR far-red light - P_fr far-red-absorbing from of phytochrome - P total phytochrome content - R red light - WL white light from fluorescent lamps