Circadian Rhythms in Stomatal Responsiveness to Red and Blue Light

Stomata of many plants have circadian rhythms in responsiveness to environmental cues as well as circadian rhythms in aperture. Stomatal responses to red light and blue light are mediated by photosynthetic photoreceptors; responses to blue light are additionally controlled by a specific blue-light photoreceptor. This paper describes circadian rhythmic aspects of stomatal responsiveness to red and blue light in Vicia faba. Plants were exposed to a repeated light:dark regime of 1.5:2.5 h for a total of 48 h, and because the plants could not entrain to this short light:dark cycle, circadian rhythms were able to "free run" as if in continuous light. The rhythm in the stomatal conductance established during the 1.5-h light periods was caused both by a rhythm in sensitivity to light and by a rhythm in the stomatal conductance established during the preceding 2.5-h dark periods. Both rhythms peaked during the middle of the subjective day. Although the stomatal response to blue light is greater than the response to red light at all times of day, there was no discernible difference in period, phase, or amplitude of the rhythm in sensitivity to the two light qualities. We observed no circadian rhythmicity in net carbon assimilation with the 1.5:2.5 h light regime for either red or blue light. In continuous white light, small rhythmic changes in photosynthetic assimilation were observed, but at relatively high light levels, and these appeared to be attributable largely to changes in internal CO2 availability governed by stomatal conductance.     

Predictors of light-limited growth and competition of phytoplankton in a well-mixed water column

Based on a model of light limited growth, Huisman and Weissing found that in a well mixed water column with constant light supply (energy reaching the water surface), equilibrium growth and competition of phytoplankton for light can be characterised by a critical light intensity at the base of the column (I*out). The present study attempts to give a further insight into this model. We first analyse the dependence of the critical light intensity on four parameters: initial slope of the photosynthesis-intensity (p-I) curve, maximal photosynthetic rate, the light-saturated parameter Ikand specific carbon loss rate. Increases in the first two parameters tend to reduce the critical light intensity and increases in the last two tend to increase the critical light intensity. Then we analyse the performance of a model under variable light supply with a time-scale of 1 day (24 hr). Within this time-scale, the critical light intensity changes with time. However, the equilibrium growth and the outcome of competition for light can be adequately characterised by critical light extinction defined as the upper limit of total light extinction due to both biomass and non-living matter in the water column. Under constant light supply, a critical light intensity uniquely corresponds to a critical light extinction. Therefore, critical light extinction can be utilised to predict the equilibrium growth and the outcome of competition under both constant and variable light supply. By changing the maximal light supply at noon, seasonal succession of species composition of communities is investigated. The possible effect of two typical photoresponses, photoadaptation and photoinhibition, on growth and competiton are discussed. Copyright 1999 Academic Press.     

The Light Requirement for Different Steps in the Development of Chloroplasts in Excised Wheat Roots

For the formation of chloroplasts in excised wheat roots blue light is necessary, but red light enhances the blue light effect. Intensity-response relationships and action spectra have been determined for the specific blue light effect as well as for the effect of red light when given before or after blue light. The longest wave length giving a blue light effect is about 500 nm. The spectrum shows a peak at about 450 nm, and a considerable effect extends down to 368 nm, the shortest wave length tested. The photoreceptor mechanism for the “blue reaction” is thought to be related to that of certain phototropisms and chloroplast movements, and to that involved in carotenoid synthesis in certain fungi. The active pigment is thought to be either a carotenoid or a flavoprotein, probably the latter. The effect of red light shows similar action spectra whether the red light is given before or after the blue, and they are consistent with protochlorophyll absorption. However, much less light is needed to produce a detectable response when the red light is given before the blue light than after. Because of the similarity of the spectra phytochrome may also be involved in the response to red light when given before blue. Far-red reversal has not been tested. The results are discussed in relation to the development of chloroplasts in leaves.     

Effects of different LED sources on the growth and nitrogen metabolism of lettuce

Using a light-emitting diode (LED) as the light source, the effects of eight different light treatments [white light (control, W), purple light (P), blue light (B), red light (R), green light (G), yellow light (Y), red–blue light in a 9:1 ratio (9R/1B), and red–blue light in a 4:1 ratio (4R/1B)] on the growth, quality and nitrogen metabolism of lettuce were studied. The results showed that compared with the white light, the purple light, blue light, red light, and the red-blue light combination could all increase the biomass of the aboveground part of lettuce to various degrees, while green light and yellow light inhibited lettuce growth. Under blue light, the contents of soluble protein and flavonoid in lettuce were the highest; under red light, the soluble sugar content was the highest, while the contents of soluble protein, free amino acids, and vitamin C (VC) were relatively higher under the 4R/1B light condition. Compared with white light, the sources of purple, blue, and red lights as well as the red–blue light combination all significantly reduced nitrate accumulation in lettuce, and the activities of the nitrogen (N) metabolism-related enzymes such as nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, and glutamate dehydrogenase were increased to varying degrees. In contrast, the contents of nitrate and ammonium N were significantly accumulated in lettuce under green light, and the activities of relative enzymes were significantly reduced. Therefore, the purple light, blue light, and red–blue combination light sources could promote N assimilation and improve the aboveground biomass accumulation in lettuce by improving the activity of the N metabolism-related enzymes in lettuce. Particularly under the 4R/1B light source, the biomass, soluble protein, VC, and total amino acid content were rather high in lettuce, which indicated that the 4R/1B light source could better effectively improve the nutritional quality and promote the growth of lettuce, while yellow light and green light are not suitable to serve as direct sources in a plant factory. These results provide a certain theoretical basis for the regulation of the light environment in cultivation facilities.     

Light quality influences adventitious shoot production from cotyledon explants of lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L.)

Summary The influence of light quality on competence and determination for organogenesis was investigated using lettuce cotyledon explants. Lettuce seedlings from four genotypes were germinated in the dark or under white, red, or blue light. Cotyledon explants were excised and cultured on a shoot-inducing medium for 28 d under white light. Germination in the dark reduced shoot numbers, suggesting that light improves the competence of explants for organogenesis. When explants from seedlings germinated under white light were cultured under different light qualities, blue was found to inhibit shoot production while red light either promoted production or had no effect on shoot number compared to controls. Treatment with blue plus red light failed to overcome the inhibition by blue light. To ascertain the temporal responses of explants to light quality, they were cultured under red or blue light prior to transfer to the alternate treatment. Exposure to blue light within 7 d of excision permanently reduced explant competence for organogenesis. Exposure after this time had a minimal effect. These results suggest that both phytochrome and cryptochrome can regulate shoot production from lettuce cotyledon explants and blue light can only inhibit organogenesis, in lettuce, during a relatively small developmental window.     

Growth towards light as an adaptation to high light conditions in Chara branches

Growth of plants or plant organs towards more light is commonly interpreted as an adaptation to low light conditions. Here, we show for the first time, in a study of charophyte branches, a growth-based orientation towards light functioning as a mechanism to protect the plant from excessive light. Two Chara species were exposed to five different intensities of photosynthetically active radiation and species traits and pigmentation were measured. Branches of plants exposed to higher light intensities were convergent and pointed steeply upwards, whereas those exposed to lower light intensities grew nearly straight and were less inclined. Only branches that increased in length during the experiments reacted to differences in light intensity. This indicates that branch orientation is determined by a light-dependent growth reaction. Orientation of charophyte branches towards light is accompanied by a decrease in chlorophyll a (Chla) content and a lower Chla : carotenoid ratio, which clearly indicates that the plant is taking protective measures against potentially damaging excess light conditions. We suggest that the growth-based orientation of Chara branches towards light may protect sexual organs, which grow on adaxial branch sides, from light damage. In addition, the upward orientation of branches might lead to increased light transmission within dense charophyte beds, thus enabling an enhanced gross production.     

Amino acid sequence of the phosphorylation site of bovine cardiac myosin light chain

Amino acid sequences of peptides containing the phosphorylation site of bovine cardiac myosin light chain (L2) were determined. The site was localized to a serine residue in the tentative amino terminus of the light chain and is homologous to phosphorylation sites in other myosin light chains. Phosphorylation of bovine cardiac light chain by chicken gizzard myosin light chain kinase was Ca2+-calmodulin dependent. Kinetic data gave a Km of 107; microM and a Vmax of 23.6 mumol min-1 mg-1. In contrast to what has been observed with smooth muscle light chains, neither the phosphorylation site fragment of the cardiac light chain nor a synthetic tetradecapeptide containing the phosphorylation site were effectively phosphorylated by the chicken gizzard kinase. Phosphorylation of cardiac myosin light chains by chicken gizzard myosin light chain kinase, therefore, requires other regions of the light chain in addition to a phosphate acceptor site.     

Light quality influences the polyamine content of lettuce (Lactuca sativa L.) cotyledon explants during shoot production <Emphasis Type="Italic">in vitro</Emphasis>

Light quality has previously been shown to influence morphogenesis in lettuce cotyledon explants, with white or red light promoting adventitious shoot production, and blue light inhibiting it. Endogenous polyamine (PA) concentrations were compared between explants cultured under different light qualities. Explants cultured under white or red light accumulated PAs during shoot primordia production, with a 5.6-fold increase compared to initial concentrations under white light, and 6.7-fold increase under red light. These results suggest polyamines are involved in the formation of shoot primordia. After 18 days in culture PA concentrations decreased under white light, and to a lesser extent under red light, signaling a shift in polyamine metabolism that correlates with shoot expansion, which occurs more readily under white light. Explants cultured under blue light accumulated polyamines for the first 7 days, to a level 1.3 times greater than initial values, followed by a gradual decline during the remainder of the culture period. Explants cultured under blue light also contained a greater proportion of PCA-insoluble conjugated PAs, compared to explants under white or red light, which contained greater proportions of free or PCA-soluble conjugated polyamines. The ratio of putrescine to spermidine was also different with a lower Put:Spd ratio being associated with shoot production under white or red light, and higher Put:Spd ratio being associated with culture under blue light.     

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.     

Measurement of light gradients and spectral regime in plant tissue with a fiber optic probe

A method is described in which light gradients and spectral regime can be measured within plant tissue using fiber optics. A fiber optic probe was made by modifying a single optical fiber (200 μm diameter) so that it had a light harvesting end that was a truncated tip 20–70 μm in diameter. The probe was a directional sensor with a half-band acceptance angle of 17–20°. Light measurements were made as the fiber optic probe was driven through plant tissue by a motorized micromanipulator, and the light that entered the fiber tip was piped to a spectroradiometer. By irradiating green leaf tissue of the succulent Crassula falcata L. with collimated light and inserting the probe from different directions, it was possible to measure light quality and quantity at different depths. Collimated light was scattered completely by the initial 1.0 mm of leaf tissue, which also greatly attenuated all light except the green and far-red. Light scatter contributed significantly to light quantity and had a pronounced spectral structure. Immediately beneath the irradiated surface the amount of light at 550 nm was 1.2 times that of the incident light. The light gradient declined rapidly to 0.5 times incident light at 1.4 mm depth. In contrast, the amount of light at 750 nm increased during the initial 0.5 mm to 2.9 times incident light and then declined linearly to 0.5 times incident light at the dark side of the leaf (4.5 mm). The implications of the magnitude of the contribution of light scatter to the light gradient is also discussed.     

Stomata from npq1, a zeaxanthin-less Arabidopsis mutant, lack a specific response to blue light

The Arabidopsis mutant npq1, which cannot accumulate zeaxanthin because of a defective violaxanthin deepoxidase, was used to investigate the role of zeaxanthin in the stomatal response to blue light. Neither dark-adapted nor light-treated guard cells or mesophyll cells of the npq1 mutant contained detectable zeaxanthin. In contrast, wild-type guard cells had a significant zeaxanthin content in the dark and accumulated large amounts of zeaxanthin when illuminated. The well-documented red light enhancement of blue light-stimulated stomatal opening, in which increasing fluence rates of background red light result in increased response to blue light, was used to probe the specific blue light response of Arabidopsis stomata. Stomata from the npq1 mutant did not have a specific blue light response under all fluence rates of background red light tested. On the other hand, stomata from leaves of hy4 (cry 1), an Arabidopsis mutant lacking blue light-dependent inhibition of hypocotyl elongation, had a typical enhancement of the blue light response by background red light. The lack of a specific blue light response in the zeaxanthinless npq1 mutant provides genetic evidence for the role of zeaxanthin as a blue light photoreceptor in guard cells.     

Redox Processes in the Blue Light Response of Guard Cell Protoplasts of Commelina communis L

Guard cell protoplasts from Commelina communis L. illuminated with red light responded to a blue light pulse by an H⁺ extrusion which lasted for about 10 minutes. This proton extrusion was accompanied by an O₂ uptake with a 4H⁺ to O₂ ratio. The response to blue light was nil in darkness without a preillumination period of red light and increased with the duration of the red light illumination until about 40 minutes. However, acidification in response to a pulse of blue light was obtained in darkness when external NADH (1 millimolar) was added to the incubation medium, suggesting that redox equivalents necessary for the expression of the response to blue light in darkness may be supplied via red light. In accordance with this hypothesis, the photosystem II inhibitor 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (10 micromolar) decreased the acidification in response to blue light more efficiently when it was added before red light illumination than before the blue light pulse. In the presence of hexacyanoferrate, the acidification in response to a blue light pulse was partly inhibited (53% of control), suggesting a competition for reducing power between ferricyanide reduction and the response to blue light.     

Effect of blue light‐emitting diode light and antioxidant potential in a somatic cell

Light is an indispensable part of routine laboratory work in which conventional light is generally used. Light‐emitting diodes (LEDs) have come to replace conventional light, and thus could be a potent target in biomedical studies. Since blue light is a major component of visible light wavelength, in this study, using a somatic cell from the African green monkey kidney, we assessed the possible consequences of the blue spectra of LED light in future animal experiments and proposed a potent mitigation against light‐induced damage. COS‐7 cells were exposed to blue LED light (450 nm) and the growth and deoxyribonucleic acid (DNA) damage were assessed at different exposure times. A higher suppression in cell growth and viability was observed under a longer period of blue LED light exposure. The number of apoptotic cells increased as the light exposure time was prolonged. Reactive oxygen species (ROS) generation was also elevated in accordance to the extension of light exposure time. A comparison with dark‐maintained cells revealed that the upregulation of ROS by blue LED light plays a significant role in causing cellular dysfunction in DNA in a time‐dependent manner. In turn, antioxidant treatment has been shown to improve cell growth and viability under blue LED light conditions. This indicates that antioxidants have potential against blue LED light‐induced somatic cell damage. It is expected that this study will contribute to the understanding of the basic mechanism of somatic cell death under visible light and maximize the beneficial use of LED light in future animal experiments.     

Light inhibition of internode elongation in green plants

The clongation of the first internode of fully greenVigna sinensis L. is inhibited by white light (W). This inhibition is fluence-rate dependent between 0 and 70 Wm^–2. The kinetics of elongation rate in the light after darkness were investigated with linear displacement transducers. The internode elongation rate does not exhibit any endogenous rhythm. A rapid inhibition occurs during the first 2 or 3 h after the onset of light, and a second type of inhibition (slow reaction) increases from the beginning to the 8th hour of light. The rapid inhibition is not fluence-rate dependent between 20 and 70 Wm^–2, but the slow reaction is. There is no rapid inhibition in a low fluence rate white light to high fluence rate white light transition, only the slow reaction is observed. The responses to different wavebands, i.e., blue light (B), yellow and green light (YG), and red light (R), are the same for the two inhibition reactions. Each waveband used separately does not reproduce the full effect observed in W. Results show a stimulation with B, a greater inhibition activity with YG than with R, and a synergistic action of B and R which when given together lead to an inhibition similar to that obtained in W. Plants returned from the light to darkness progressively recover a high elongation rate without any latent period. The W light regulating internode elongation rate is mainly perceived by the growing internode itself.Abbreviations B blue light - D darkness - F far-red light - HW high fluence rate white light - LW low fluence rate white light - R red light - W white light - YG yellow and green light     

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.     

Light‐induced stabilization of ACS contributes to hypocotyl elongation during the dark‐to‐light transition in Arabidopsis seedlings

Hypocotyl growth during seedling emergence is a crucial developmental transition influenced by light and phytohormones such as ethylene. Ethylene and light antagonistically control hypocotyl growth in either continuous light or darkness. However, how ethylene and light regulate hypocotyl growth, including seedling emergence, during the dark‐to‐light transition remains elusive. Here, we show that ethylene and light cooperatively stimulate a transient increase in hypocotyl growth during the dark‐to‐light transition via the light‐mediated stabilization of 1‐aminocyclopropane‐1‐carboxylic acid (ACC) synthases (ACSs), the rate‐limiting enzymes in ethylene biosynthesis. We found that, in contrast to the known inhibitory role of light in hypocotyl growth, light treatment transiently increases hypocotyl growth in wild‐type etiolated seedlings. Moreover, ACC, the direct precursor of ethylene, accentuates the effects of light on hypocotyl elongation during the dark‐to‐light transition. We determined that light leads to the transient elongation of hypocotyls by stabilizing the ACS5 protein during the dark‐to‐light transition. Furthermore, biochemical analysis of an ACS5 mutant protein bearing an alteration in the C‐terminus indicated that light stabilizes ACS5 by inhibiting the degradation mechanism that acts through the C‐terminus of ACS5. Our study reveals that plants regulate hypocotyl elongation during seedling establishment by coordinating light‐induced ethylene biosynthesis at the post‐translational level. Moreover, the stimulatory role of light on hypocotyl growth during the dark‐to‐light transition provides additional insights into the known inhibitory role of light in hypocotyl development.     

Effect of Low-intensity Red and Far-red Light and High-intensity White Light on the Flowering Response of the Long-day Plant Lemna gibba G3

The long-day plant Lemna gibba L., strain G3 exhibits a relatively low sensitivity to short, white-light interruptions given during the dark period of a short-day cycle. However, the plants are fairly sensitive to low-intensity red light treatments given during a 15-hour dark period on the third day of a 2LD-(9L:15D)-2LD-7SD schedule. Far-red light is almost as effective as red light, and attempts to reverse the red light response with subsequent far-red light treatments have not been successful. Blue light proved to be without effect. When plants were grown on a 48-hour cycle with 15 minutes of red light every 4 hours during the dark period, the critical daylength was reduced from about 32 hours to slightly less than 12 hours.

Continuous red light induced a fairly good flowering response. However, as little as 1 hour of white light each day gave a significant improvement in the flowering response over that of the continuous red light control. White light of 600 to 700 ft-c was more effective than white light of 60 to 70 ft-c. The white light was much more effective when divided into 2 equal exposures given 8 to 12 hours apart. These results suggest an increase in light sensitivity with regard to flower induction about 8 to 10 hours after the start of the light period.

     

Circadian rhythms of locomotor activity in solitary and social species of African mole-rats (family: Bathyergidae)

Mole-rats are strictly subterranean and hardly, if ever, come into contact with external light. As a result, their classical visual system is severely regressed and the circadian system proportionally expanded. The family Bathyergidae presents a unique opportunity to study the circadian system in the absence of the classical visual system in a range of species. Daily patterns of activity were studied in the laboratory under constant temperature but variable lighting regimes in individually housed animals from 3 species of mole-rat exhibiting markedly different degrees of sociality. All 3 species possessed individuals that exhibited endogenous circadian rhythms under constant darkness that entrained to a light-dark cycle. In the solitary species, Georychus capensis, 9 animals exhibited greater activity during the dark phase of the light cycle, while 2 individuals expressed more activity in the light phase of the light cycle. In the social, Cryptomys hottentotus pretoriae, 5 animals displayed the majority of their activity during the dark phase of the light cycle and the remaining 2 exhibited more activity during the light phase of the light cycle. Finally in the eusocial Cryptomys damarensis, 6 animals displayed more activity during the light phase of the light cycle, and the other 2 animals displayed more activity during the dark phase of the light cycle. Since all three mole-rat species are able to entrain their locomotor activity to an external light source, light must reach the SCN, suggesting a functional circadian clock. In comparison to the solitary species, the 2 social species display a markedly poorer response to light in all aspects. Thus, in parallel with the sociality continuum, there exists a continuum of sensitivity of the circadian clock to light.     

The Effect of Red Irradiation on Plastid Ribosomal RNA Synthesis in Dark-grown Pea Seedlings

Dark-grown pea seedlings (Pisum sativum L.) were irradiated for a short period each day with low intensity red light (662 nm), red light immediately followed by far red light (730 nm), or far red light alone. Other plants were transferred to a white light regime (14 hours light/10 hours dark). There was no change in the amount of RNA in the tissue on a fresh weight basis after the various treatments. However, compared with dark-grown seedlings, those plants irradiated with red light showed an increase in the net RNA content per stem apex. In addition there was a two- to three-fold increase in ribosomal RNA of the etioplasts relative to the total ribosomal RNA. These increases were comparable to those found in plants grown in the white light regime. The changes were much smaller if the dark-grown plants were irradiated either with red light followed by far red light, or with far red light alone. Thus continuous light is not essential for the production of ribosomal RNA in plastids, and the levels of ribosomal RNA found in chloroplasts can also be attained in etioplasts of pea leaves in the dark provided the leaf phytochrome is maintained in its active form.