High Irradiance Response Research Articles

Phytochrome elicits the cryptic red-light signal which results in amplification of anthocyanin biosynthesis in sorghum

Anthocyanin synthesis in Sorghum bicolor Moench induced by a low-fluence response of phytochrome (phy) is multiplicatively amplified by a cryptic red-light signal (CRS) produced by red light (R). The photoreceptor for CRS and its features in CRS production were studied. (i) An action spectrum determined with a 200-s light pulse of wavelengths from 347 to 693 nm had peaks at 657 and 378 nm. (ii) The CRS-producing effect of R, even as short a pulse as 20 s, was neither suppressed by an immediately subsequent far-red light (FR) pulse nor increased by placing a dark interval of 180 s between R and FR; simultaneous FR, however, suppressed the R action in accordance with the resulting ratios of the FR-absorbing form (Pfr) to total phy. (iii) The effect of R increased with increasing fluence rate to plateau at the same fluence rate regardless of the pulse length, but the level of this plateau depended on the pulse length. (iv) The effect of R increased with increasing pulse length when compared at the same fluence, whether saturating or unsaturating; thus, no reciprocity law holds. These results indicate that the photoreceptor for CRS production is a phy, Pfr being active, which presumably shows very fast dark reversion to the R-absorbing form without absorbing FR. The possible CRS-production mechanism of the phy and its significance in the so-called R high-irradiance response of phy are discussed.

In Vivo Characterization of Chimeric Phytochromes in Yeast

Phytochromes are plant photoreceptors that play a major role in photomorphogenesis. Two members of the phytochrome family have been characterized in some detail. Phytochrome A, which controls very low fluence and high irradiance responses, is rapidly degraded in the light, forms sequestered areas of phytochrome (SAPs), and does not exhibit dark reversion in monocotyledonous seedlings. Phytochrome B mediates red/far-red reversible responses, is stable in the light, and does not form SAPs. We report on the behavior in yeast of the phytochrome apoproteins of rice PHYA, tobacco PHYB, and chimeric PHYAB and PHYBA and on the behavior of the respective holoprotein adducts after assembly with phycocyanobilin chromophore (PHY*). SAP-like formation in yeast was not observed for PHYB, but was detectable for PHYA, PHYAB, and PHYBA. Rice PHYA* did not undergo dark reversion in yeast. Surprisingly, all other tested phytochrome constructs did exhibit dark reversion, including chimeric phytochromes with a short N-terminal part of tobacco PHYB or parsley PHYA fused to rice PHYA. Furthermore, the proportion of phytochrome undergoing dark reversion and the rate of reversion were increased for both the N terminus-swapped constructs and PHYBA*. These results are discussed with respect to structure/function analysis of phytochromes A and B.

Different Regulation of β-amylase and Starch Phosphorylase by Light in Dormant and Non-Dormant Turions of Spirodela polyrhiza

Summary The influence of single red light pulses and of continuous red light on the activity of two enzymes, i.e. β-amylase (EC 3.2.1.2) and starch phosphorylase (EC 2.4.1.1), was investigated in dormant and non-dormant turions of Spirodela polyrhiza. The activity of β-amylase was induced by continuous red light but not by red light pulses in dormant turions. In non-dormant turions, however, the activity of β-amylase following a single red light pulse was two times higher than after continuous red light. This high effectiveness of a single red light pulse, as compared with continuous red, represents a novel mode of regulation of enzyme activity by light. It is suggested that a low fluence response of phytochrome is abolished by a high irradiance response elicited by continuous red light. The light-induced increase in the activity of β-amylase is due to de novo synthesis of the enzyme as shown by Western blot analysis using polyclonal antibodies raised against β-amylase from Spirodela polyrhiza. The presence of light-activable precursor was excluded by showing that cysteine added to the extraction medium did not influence the activity of β-amylase. In contrast, the activity of starch phosphorylase was induced by continuous red light in dormant turions but decreased in non-dormant turions. During the cold after-ripening treatment turions qualitatively changed their response to light, becoming light-insensitive when partly after-ripened (21 days). In all instances, a red light pulse was ineffective in inducing starch phosphorylase. Isoforms of β-amylase and starch phosphorylase were separated by non-denaturating PAGE. No special induction by light of any specific isoform of β-amylase and starch phosphorylase was found in turions. Since both enzymatic activities and their regulation by light do not correlate with the breakdown of starch, we conclude that they are not directly involved in the regulation of starch degradation in turions.

Modes of action of phytochromes

through largely unknown pathways into molecular and physiological changes that modulate growth and developPhytochrome is a family of photoreceptors whose ment. Phytochromes posses two photo-interconvertible apoproteins are encoded by divergent genes. Three forms, the R-absorbing form (Pr) and the FR-absorbing modes of action of phytochromes, very-low-fluence form (Pfr) and are synthesised in the Pr form. In recent responses (VLFR), low-fluence responses (LFR) and years it has been established that phytochromes are a high-irradiance responses (HIR), have been considered small family of photoreceptors whose apoproteins are in the literature to define the quantitative relationship encoded by diVerent genes (Clack et al., 1994; Hauser between response and predicted levels of the far-red et al., 1995). Traditionally, three ‘modes of action’ of light absorbing form of phytochrome. The results of phytochromes have been described: very-low-fluence recent experiments with phytochrome mutant and responses ( VLFR); low-fluence responses (LFR) and transgenic seedlings are consistent with the notion high-irradiance responses (HIR). Assigning particular that phytochrome B, and in some cases other stable responses to particular molecular species of phytochromes phytochromes, mediate LFR whereas phytochrome A is one of the important areas of research in the field mediates both VLFR and HIR. The mechanisms of photomorphogenesis. This review seeks to bring involved in phytochrome A inductive responses (VLFR) together data on the modes of action of phytochrome versus HIR appear to be different in some ways. with information on diVerent phytochrome molecular However, VLFR, LFR and HIR can converge to control species. the transcriptional activity of a single gene promoter. The analysis of phytochrome transduction chains is The implications of these findings for the under- currently concentrating the eVorts of many laboratories. standing of phytochrome phototransduction are Experimental approaches include the characterization of discussed. the photoreceptor molecule, in particular the biological significance of diVerent domains (Cherry et al., 1993;

The photoreceptors in the high irradiance response of plants

Several studies show that the high irradiance response (HIR) of plants is probably due to two photoreceptors. One of the photoreceptors is phytochrome, and the other is an unidentified pigment provisionally named heliochrome. One of the functions of heliochrome is the synthesis of phytochrome, using far‐red and blue radiations of high intensities, to replace the phytochrome destroyed by light. Another possible function could be an interaction of heliochrome with a substance produced by phytochrome. The data presented show that heliochrome is a pigment with different properties from phytochrome. It shows a far‐red/green reversibility. Heliochrome has been shown to participate with phytochrome in such HIRs as leaf movement in Albizzia and flowering in a long‐day plant. The first event initiated by phytochrome and by heliochrome could be the generation of a strong positive, electrostatic charge in the cell membrane.

The photoreceptors in the high irradiance response of plants

The photoreceptors in the high irradiance response of plants

The VLF loci, polymorphic between ecotypes Landsberg erecta and Columbia, dissect two branches of phytochrome A signal transduction that correspond to very‐low‐fluence and high‐irradiance responses

Phytochromes play a key role in the perception of light signals by plants. In this study, the three classical phytochrome action modes, i.e. very‐low‐fluence responses (VLFR), low‐fluence responses (LFR) and high‐irradiance responses (HIR), were genetically dissected using phyA and phyB mutants of Arabidopsis thaliana (respectively lacking phytochrome A or phytochrome B) and a polymorphism between ecotypes Landsberg erecta and Columbia. Seed germination and potentiation of greening, hypocotyl growth inhibition and cotyledon unfolding in etiolated seedlings of the ecotype Landsberg erecta showed biphasic responses to the calculated proportion of active phytochrome established by one light pulse or repeated light pulses. The first phase, i.e. the VLFR, was absent in the phyA mutant, normal in the phyB mutant (both in the Landsberg erecta background) and severely deficient in Columbia. The second phase, i.e. the LFR, was present in the phyA mutant, deficient in the phyB mutant and normal in Columbia. Under continuous far‐red light, HIR of etiolated seedlings were absent in phyA and normal in phyB and Columbia. The segregation of VLFR in recombinant inbred lines derived from a cross between Landsberg erecta and Columbia was analysed by MAPMAKER/QTL. Two quantitative trait loci, one on chromosome 2 (VLF1) and another on chromosome 5 (VLF2), were identified as responsible for the polymorphism. Phytochrome A is proposed to initiate two transduction pathways, VLFR and HIR, involving different cells and/or different molecular steps. This is the first application of the analysis of quantitative trait loci polymorphic between ecotypes to dissect transduction chains of environmental signals.

The VLF loci, polymorphic between ecotypes Landsberg erecta and Columbia, dissect two branches of phytochrome A signal transduction that correspond to very-low-fluence and high-irradiance responses.

Phytochromes play a key role in the perception of light signals by plants. In this study, the three classical phytochrome action modes, i.e. very-low-fluence responses (VLFR), low-fluence responses (LFR) and high-irradiance responses (HIR), were genetically dissected using phyA and phyB mutants of Arabidopsis thaliana (respectively lacking phytochrome A or phytochrome B) and a polymorphism between ecotypes Landsberg erecta and Columbia. Seed germination and potentiation of greening, hypocotyl growth inhibition and cotyledon unfolding in etiolated seedlings of the ecotype Landsberg erecta showed biphasic responses to the calculated proportion of active phytochrome established by one light pulse or repeated light pulses. The first phase, i.e. the VLFR, was absent in the phyA mutant, normal in the phyB mutant (both in the Landsberg erecta background) and severely deficient in Columbia. The second phase, i.e. the LFR, was present in the phyA mutant, deficient in the phyB mutant and normal in Columbia. Under continuous far-red light, HIR of etiolated seedlings were absent in phyA and normal in phyB and Columbia. The segregation of VLFR in recombinant inbred lines derived from a cross between Landsberg erecta and Columbia was analysed by MAPMAKER/QTL. Two quantitative trait loci, one on chromosome 2 (VLF1) and another on chromosome 5 (VLF2), were identified as responsible for the polymorphism. Phytochrome A is proposed to initiate two transduction pathways, VLFR and HIR, involving different cells and/or different molecular steps. This is the first application of the analysis of quantitative trait loci polymorphic between ecotypes to dissect transduction chains of environmental signals.

Seed germination of Arabidopsis thaliana phyA/phyB double mutants is under phytochrome control.

We examined the photocontrol of seed germination in the phyA/phyB double mutants of Arabidopsis thaliana seeds. Dormant phyA/phyB seeds showed a red/far-red light (R/FR)-reversible induction of seed germination. This suggests the involvement of at least one other phytochrome, phyC, D, and/or E, in controlling seed germination. We designated this spectrally active phytochrome in phyA/phyB as phyX. The full reversibility of the R-induced germination by subsequent FR pulses, and the observation that the response is reversible by FR, even after a 3-h R treatment, indicates that this phyX response belongs to the low-fluence-response type. Thus, this phyX response is functionally related to phyB-mediated responses. However, in contrast to phyB-controlled seed germination, this phyX-mediated response needs a prolonged imbibition period and exhibits reversibility kinetics different from that needed for phyB. Furthermore, this phyX response requires a prolonged irradiation time and shows a fluence rate response dependency, showing a similarity to the high irradiance response of photomorphogenesis. Thus, phyX, with regard to its control of seed germination, is a functionally new phytochrome that shares some characteristics of both phyA- and phyB-mediated responses.

Phytochrome-regulated Starch Degradation in Germinating Turions of Spirodela polyrhiza

— Turions of the duckweed Spirodela polyrhiza contain about 70% starch on the dry weight basis. The rate of starch degradation in nondormant turions was highest in continuous red light (cR) followed by continuous blue light (cB), whereas continuous far red light (cFR) is almost ineffective. Continuous R could be substituted by repeated R pulses; this effect was not photoreversible by FR pulses applied after hourly R pulses. This suggested that R-dependent high irradiance response is the mode of phytochrome action in mediating starch degradation. Comparing the mode of phytochrome action with that of phytochrome mutants of other plants it can be assumed that phytochrome B is the photoreceptor involved. Application of the translation inhibitor cycloheximide immediately stopped the phytochrome-dependent starch degradation demonstrating the involvement of de novo synthesis of proteins.

Antagonistic but complementary actions of phytochromes A and B allow seedling de-etiolation.

Using dichromatic radiation, we show that the actions of phytochromes A and B (phyA and phyB) in Arabidopsis thaliana are antagonistic in mediating red and far-red radiation effects on seedling de-etiolation and yet act in a complementary manner to regulate de-etiolation, irrespective of spectral composition. At low phytochrome photoequilibria inhibition of hypocotyl extension was strong, because of the action of a far-red high-irradiance response mediated by phyA. At high phytochrome photoequilibria inhibition of hypocotyl extension was also strong, because of the action of phyB. At intermediate photoequilibria hypocotyl inhibition was less strong. In their natural environment, this dual action will strongly retard hypocotyl growth and promote cotyledon opening and expansion both in open daylight and under dense vegetation. Overlapping action by phyA and phyB will substantially promote de-etiolation in sparse vegetation. The antagonistic and complementary actions of phyA and phyB, therefore, allow the optimum regulation of seedling growth after emergence from the soil.

Sucrose control of phytochrome A signaling in Arabidopsis.

The expression of the Arabidopsis plastocyanin (PC) gene is developmentally controlled and regulated by light. During seedling development, PC gene expression is transiently induced, and this induction can be repressed by sucrose. In transgenic seedlings carrying a PC promoter-luciferase fusion gene, the luciferase-induced in vivo luminescence was similarly repressed by sucrose. From a mutagenized population of such transgenic seedlings, we selected for mutant seedlings that displayed a high luminescence level when grown on a medium with 3% sucrose. This screening of mutants resulted in the isolation of several sucrose-uncoupled (sun) mutants showing reduced repression of luminescence by sucrose. Analysis of the sun mutants revealed that the accumulation of PC and chlorophyll a/b binding protein (CAB) mRNA was also sucrose uncoupled, although the extent of uncoupling varied. The effect of sucrose on far-red light high-irradiance responses was studied in wild-type, sun1, sun6, and sun7 seedlings. In wild-type seedlings, sucrose repressed the far-red light-induced cotyledon opening and inhibition of hypocotyl elongation. sun7 seedlings showed reduced repression of these responses. Sucrose also repressed the far-red light-induced block of greening in wild-type seedlings, and both sun6 and sun7 were affected in this response. The results provide evidence for a close interaction between sucrose and light signaling pathways. Moreover, the sun6 and sun7 mutants genetically identify separate branches of phytochrome A-dependent signal transduction pathways.

Photocontrol of anthocyanin biosynthesis in tomato.

Juvenile anthocyanin biosynthesis has been studied in dark-grown seedlings of tomato (Lycopersicon esculentum Mill.) wild types (WTs) and photomorphogenic mutants. During a subsequent 24-hr period of monochromatic irradiation at different fluence rates of red light (R) the fluence-rate response relationships for induction of anthocyanin in all the WTs are similar, yet complex, showing a response at low fluence rates (LFRR) followed by a fluence rate-dependent high irradiance response (HIR). In the hypocotyl this response is restricted to the sub-epidermal layer of cells. The high-pigment-1 (hp-1) mutant exhibits a strong amplification of both response components. Theatroviolacea (atv) mutant shows strongest amplification of the HIR component. In contrast, a transgenic line overexpressing an oat phytochrome A gene (PHYA3 (+)) shows a most dramatic amplification of the LFRR component. The far-red light (FR)-insensitive (fri) mutant, deficient in phytochrome A (phyA), lacks the LFRR component whilst retaining a normal HIR. The temporarily R-insensitive (tri) mutant, deficient in phytochrome B1 (phyB1) retains the LFRR, but lacks the HIR. Thehp-1,fri andhp-1,tri double mutant, exhibit amplified, yet qualitatively similar responses to the monogenicfri andtri mutants. Thefri,tri double mutant lacks both response components in R, but a residual response to blue light (B) remains. Similarly, theaurea (au) mutant deficient in phytochrome chromophore biosynthesis and presumably all phytochromes, lacks both response components in the R and FR regions of the spectrum. Experiments at other wavelengths demonstrate that while there is only a small response in the FR spectral region (729 nm) in tomato, there is an appreciable HIR response in the near FR at 704 nm, which is retained in thetri mutant. This suggests that the labile phyA pool participates in the HIR at this wavelength. The intense pigmentation (Ip) mutant appears to be specifically deficient in the B1 induced anthocyanin biosynthesis. Adult plants, grown under fluorescent light/dark cycles, show a reduction of anthocyanin content of young developing leaves upon application of supplemtary or end-of-day FR. The involvement of different phytochrome species in anthocyanin biosynthesis based on micro-injection studies into theau mutant and studies using type specific phytochrome mutants is discussed.

Phytochrome Control of Anthocyanin Biosynthesis in Tomato Seedlings: Analysis Using Photomorphogenic Mutants

AbstractAnthocyanin biosynthesis has been studied in hypocotyls and whole seedlings of tomato (Lycoperskon esculentum Mill.) wild types (WTs) and photomorphogenic mutants. In white light (WL)/dark (D) cycles the fri1 mutant, deficient in phytochrome A (phyA), shows an enhancement of anthocyanin accumulation, whereas the tri1 mutant, deficient in phytochrome Bl (phyBl) has a WT level of anthocyanin. Under pulses of red light (R) or R followed by far‐red light (FR) given every 4 h, phyA is responsible for the non‐R/FR reversible response, whereas phyBl is partially responsible for the R/FR reversible response. From R and blue light (B) pretreatment studies, B is most effective in increasing phytochrome responsiveness, whereas under R itself it appears to be dependent on the presence of phyBl. Anthocyanin biosynthesis during a 24 h period of monochromatic irradiation at different flu‐ence rates of 4 day‐old D‐grown seedlings has been studied. At 660 nm the fluence rate‐response relationships for induction of anthocyanin in the WT are similar, yet complex, showing a low fluence rate response (LFRR) and a fluence rate‐dependent high irradiance response (HIR). The high‐pigment‐1 (hp‐1) mutant exhibits a strong amplification of both the LFRR and HIR. The fri1 mutant lacks the LFRR while retaining a normal HIR. In contrast, a transgenic tomato line overexpressing the oat PHYA3 gene shows a dramatic amplification of the LFRR. The tri1 mutant, retains the LFRR but lacks the HIR, whereas the fri1, tri1 double mutant lacks both components. Only an LFRR is seen at 729 nm in WT; however, an appreciable HIR is observed at 704 nm, which is retained in the tri1 mutant and is absent in the fri1 mutant, indicating the labile phyA pool regulates this response component.

A 146 bp fragment of the tobacco Lhcb1*2 promoter confers very-low-fluence, low-fluence and high-irradiance responses of phytochrome to a minimal CaMV 35S promoter.

The occurrence of very-low-fluence responses (VLFR), low-fluence responses (LFR) and high-irradiance responses (HIR) of phytochrome was investigated for the expression of the gene of beta-glucuronidase (gusA) under the control of the tobacco Lhcb1*2 promoter, in etiolated transgenic tobacco seedlings. The activity of beta-glucuronidase (GUS) showed biphasic responses to the calculated proportion of Pfr provided by light pulses. The first phase (i.e. the VLFR) showed a maximum for Pfr levels characteristic of far-red light. The second phase (i.e. the LFR) was observed at higher Pfr levels and was reversible by far-red light pulses. The strong effect of continuous far-red light (i.e. HIR) was fluence-rate-dependent and could not be replaced either by hourly pulses of the same spectral composition and total fluence or by very low fluences of red light. Deletion of the Lhcb1*2 promoter to -453 caused little loss of GUS activity. The -453 to -31, -270 to -31 and -176 to -31 fragments of the Lhcb1*2 promoter conferred proportionally normal VLFR, LFR and HIR to a truncated (-46 to +8) CaMV 35S minimal promoter. This is the first demonstration of the presence of three phytochrome action modes in the control of the transcriptional activity of a single gene. The cis-acting regulatory elements necessary for VLFR, LFR and HIR are present in a 146 bp fragment of the tobacco Lhcb1*2 promoter.

Physiological characterization of exaggerated-photoresponse mutants of tomato

Summary Four monogenic mutants in tomato ( Lycopersicon esculentum Mill.), i.e. three recessive mutations, high-pigment-1 ( hp-1 ), high-pigment-2 ( hp-2 ), and atroviolacea ( atv ), and one dominant mutation, Intense pigmentation ( Ip ), were used in this study. These mutants all show exaggerated photoresponses during deetiolation and seedlings having shorter hypocotyls and higher anthocyanin levels. The hp-1 and hp-2 have higher chlorophyll levels in immature fruit, giving them a dark green colour. Spectrophotometrical and immunological analyses of phytochrome A and phytochrome B revealed no differences between the mutants and the wild types (WTs), suggesting that the mutants are not photoreceptor mutants. Both hp-1 and hp-2 accumulate high levels of anthocyanin in continuous blue (B) and red (R) broad-band light. In contrast, atv has a WT level of anthocyanin in B and an exaggerated response in R. The Ip mutant has the opposite response: a WT level of anthocyanin in R and an exaggerated response in B. In B and R pretreat-ment studies, all mutants show an enhanced R/far-red light (FR)-reversible response compared with WT, but the Ip mutant shows a preferentially enhanced response in B. The hp-1 mutant exhibits a strong amplification of both the low fluence rate response and high irradiance response components of anthocyanin biosynthesis in red light. The atv mutant shows the strongest amplification of the HIR component. The Ip mutant exhibits an exaggerated anthocyanin response in B. All four mutants exhibit a normal elongation response to supplementary FR during the daily photoperiod.

High-irradiance responses induced by far-red light in grass seedlings of the wild type or overexpressing phytochrome A

The occurrence of phytochrome-mediated highirradiance responses (HIR), previously characterised largely in dicotyledonous plants, was investigated in Triticum aestivum L., Zea mays L., Lolium multiflorum Lam. and in both wild-type Oryza sativa L. and in transgenic plants overexpressing oat phytochrome A under the control of a 35S promoter. Coleoptile growth was promoted (maize, ryegrass) or inhibited (wild-type rice) by continuous far-red light (FRc). However, at equal fluences, hourly pulses of far-red light (FRp) were equally effective, indicating that the growth responses to FRc were not true HIR. In contrast, in maize and rice, FRc increased anthocyanin content in the coleoptile in a fluence-rate dependent manner. This response was a true HIR as FRp had reduced effects. In maize, anthocyanin levels were significantly higher under FRc than under continuous red light. In rice, overexpression of phytochrome A increased the inhibition of coleoptile growth and the levels of anthocyanin under FRc but not under FRp or under continuous red light. The effect of FRc was fluence-rate dependent. In light-grown rice, overexpression of phytochrome A reduced leaf-sheath length, impaired the response to supplementary far-red light, but did not affect the response to canopy shade-light. In grasses, typical HIR, i.e. fluence-rate dependent responses showing reciprocity failure, can be induced by FRc. Under FRc, overexpressed phytochrome A operates through this action mode in transgenic rice.

Ultraviolet killing and photoreactivation of Phycomyces spores

Vegetative spores of the zygomycete fungus Phycomyces blakesleeanus were used to investigate UV-killing and photoreactivation. Irradiation with far-UV light generated shouldered exponential survival curves. Postirradiation with near-UV, blue or white light caused photoreactivation, i.e. increase of the survival rate. Photon fluenceresponse curves for photoreactivation, which were generated for selected wavelengths between 350 and 610 nm of light, were generally monophasic, while they were biphasic when 385-nm light was used. At the latter wavelength one could distinguish a low-irradiance response and a high-irradiance response. A 7-point action spectrum for photoreactivation displayed a maximum at 385 nm indicating that the DNA-photolyase of Phycomyces contains as chromophores 5,10-methenyltetrahydrofolate (MTHF) and FADH 2. Spores which were obtained from dark-grown Phycomyces stocks showed no photoreactivation in response to low-irradiance light, i.e. exposures shorter than 4 min. For exposures longer than 4 min (high-irradiance response) photoreactivation did occur. The latter response was apparently due to newly induced DNA-photolyase, which is presumably absent in spores of dark-grown stocks. Two light-insensitive mutants, C47 madA and C111 madB, which are defective for phototropism and photodifferentiation, lacked photoreactivation after low and high-irradiance conditions. This finding, too, indicated that the DNA-photolyase is under stringent near-UV/blue-light control and is not expressed in darkness.

Light-driven movements of the trifoliate leaves of bean ( Phaseolus vulgaris L.). Spectral and functional analysis

Summary The light-driven responses of the terminal leaflet of bean were analyzed spectrally and functionally. Laminar elevation increases rapidly in response to continuous overhead exposure of its pulvinus to blue light. This response is enhanced in its early stages by simultaneous exposure to red light. The pulvinus responds similarly to continuous overhead unmixed red, or far-red light, albeit at much lower rates. The response to overhead red, alone, or during enhancement of the response to blue, was not affected by simultaneous far-red. However, the response to blue alone, or enhanced by mixture with red, was partially inhibited by simultaneous exposure to far-red. The results suggest that the response to blue resulted mostly from a blue-absorbing pigment system, but may involve some absorption by phytochrome, while responses to red or far-red, with and without blue, may be mediated by high-irradiance responses of phytochrome. Functional differences between the responses to red and blue become apparent when the abaxial (lower), or lateral sectors of the pulvinus are exposed to them, separately and in combination. These differences suggest that red controls the photonastic unfolding of the pulvinus, whereas blue controls its phototropic responses. These responses co-exist in the same tissue, but are separate and additive.

Phytochrome A and phytochrome B mediate the hypocotyl-specific downregulation of TUB1 by light in arabidopsis.

Arabidopsis contains six alpha-tubulin and nine beta-tubulin genes that are expressed in a tissue-specific and developmentally regulated manner. We analyzed the effects of light on tubulin mRNA abundance in Arabidopsis seedlings using RNA gel blot hybridizations and gene-specific probes. Transcript levels of all 15 tubulin genes were decreased by continuous white light, although to different degrees. Detailed analysis was performed with the beta-tubulin TUB1 gene. The transcript level of TUB1 was high in etiolated seedlings and decreased to approximately 20% of the dark mRNA level after 2 to 6 hr of white light treatment. We showed that this downregulation requires high-irradiance light treatment and that multiple photoreceptors are involved. In particular, using phytochrome mutants and narrow wave band light, we demonstrated that both the phytochrome A (phyA)-mediated far-red light high-irradiance response and the phytochrome B (phyB)-mediated red light high-irradiance response are involved in the downregulation of TUB1 expression by white light. Histochemical analysis of transgenic plants expressing a TUB1-beta-glucuronidase chimeric transgene indicated that the downregulation observed only in hypocotyls and not in roots is controlled transcriptionally.