Blue- and red-light action in photoorientation of chloroplasts in Adiantum protonemata

An action spectrum for the low-fluencerate response of chloroplast movement in protonemata of the fern Adiantum capillus-veneris L. was determined using polarized light vibrating perpendicularly to the protonema axis. The spectrum had several peaks in the blue region around 450 nm and one in the red region at 680 nm, the blue peaks being higher than the red one. The red-light action was suppressed by nonpolarized far-red light given simultaneously or alternately, whereas the bluelight action was not. Chloroplast movement was also induced by a local irradiation with a narrow beam of monochromatic light. A beam of blue light at low energy fluence rates (7.3·10^-3-1.0 W m^-2) caused movement of the chloroplasts to the beam area (positive response), while one at high fluence rates (10 W m^-2 and higher) caused movement to outside of the beam area (negative response). A red beam caused a positive response at fluence rates up to 100 W m^-2, but a negative response at very high fluence rates (230 and 470 W m^-2). When a far-red beam was combined with total background irradiation with red light at fluence rates causing a low-fluence-rate response in whole cells, chloroplasts moved out of the beam area. When blue light was used as background irradiation, however, a narrow far-red beam had no effect on chloroplast distribution. These results indicate that the light-oriented movement of Adiantum chloroplasts is caused by red and blue light, mediated by phytochrome and another, unidentified photoreceptor(s), respectively. This movement depends on a local gradient of the far-red-absorbing form of phytochrome or of a photoexcited blue-light photoreceptor, and it includes positive and negative responses for both red and blue light.Abbreviations BL blue light - FR far-red light - Pfr far-red-absorbing form of phytochrome - Pr red-absorbing form of phytochrome - R red light - UV ultraviolet     

Dual effect of phytochrome A on hypocotyl growth under continuous red light

The role of phytochrome A in the control of hypocotyl growth under continuous red light (Rc) was investigated using phyA and phyB mutants of Arabidopsis thaliana, which lack phytochrome A (phyA) or phytochrome B (phyB), respectively, and transgenic seedlings of Nicotiana tabacum overexpressing Avena phyA, compared to the corresponding wild type (WT). In WT seedlings of A. thaliana, hypocotyl growth inhibition showed a biphasic response to the fluence rate of Rc, with a brake at 10^?2μmol m^?2 s^?1. At equal total fluence rate, hourly pulses of red light caused slightly more inhibition than Rc. The response to very low fluences of continuous or pulsed red light was absent in the phyA and phyA phyB mutants and present in the phyB mutant. The second part of the response was steeper in the phyA mutant than in the WT but was absent in the phyB mutant. In WT tobacco the response to Rc was biphasic. Overexpression of Avena phyA enhanced the response only at very low fluence rates of Rc (< 10^?2μmol m^?2 s^?1). In both species, the effect of hourly pulses of far-red light was similar to the maximum inhibition observed in the first phase of the response to Rc. Using reciprocity failure (i.e. higher inhibition under continuous than pulsed light) as the operational criterion, a ‘true’ high-irradiance reaction occurred under continuous far-red light but not under Rc or red plus far-red light mixtures. Native and overexpressed phyA are proposed to mediate very low fluence responses under Rc. In WT A. thaliana, this effect is counteracted by a negative action of phyA on phyB-mediated low-fluence responses.     

Phytochrome modulation of blue-light-induced phototropism

Red light enhances hypocotyl phototropism toward unilateral blue light through a phytochrome‐mediated response. This study demonstrates how the phytochromes modulate blue‐light‐induced phototropism in the absence of a red light pre‐treatment. It was found that phytochromes A, B, and D have conditionally overlapping functions in the promotion of blue‐light‐induced phototropism. Under very low blue light intensities (0.01 µmol m^?2 s^?1) phyA activity is necessary for the progression of a normal phototropic response, whereas above 1.0 µmol m^?1 s^?2 phyB and phyD have functional redundancy with phyA to promote phototropism. PhyA also contributes to attenuation of phototropism under high fluence rates of unilateral blue light, which was previously shown to be dependent on the phototropins and cryptochromes. From these results, it appears that phytochromes are required to develop a robust phototropic response under low fluence rates, whereas under high irradiances where phototropism may be less important, phyA suppresses phototropism.     

EFFECTS OF PHOTON FLUENCE RATE AND LIGHT SPECTRUM COMPOSITION ON GROWTH,PHOTOSYNTHESIS AND PIGMENT RELATIONS IN GRACILARIA SP.1

The optimal photon fluence rate for growth of tha llus tips of Gracilaria sp. was low (about 100 μE·^–2·¹); higher photon fluence rates inhibited growth. Both phycoerythrin (PE) and chlorophyll (chl) contents decreased with increasing photon fluence rates (up to 100 μE·–m^–2s^–1) in a fashion inverse to the growth response. Chl/PE ratios varied directly as the growth response over a larger photon fluence rate range. The peak chl/PE ratios were obtained at a photon fluence rate optimal for growth, suggesting that this parameter may be used to estimate in situ growth rates. A low compensation point (about 7 μE·^–2s^–1) was observed for low light (15 μE·^–2s^–1) grown plants. This compensation point was also obtained for growth in the long–term (5–6 weeks) experiments. Plants grown at 60 and 140 μE·^–2s^–1 showed higher light compensation and saturation points, suggesting that the variations in pigment composition found between the different treatments determine the photosynthetic responses at sub–optimal photon fluence rates. Photosynthetic rates at light saturation were the same, on a biomass basis, for plants grown at the various photon fluence rates. Thus, the photosynthetic dark reactions were not influenced by previous light regimes. It is suggested that maximal photosynthetic rates expressed on a biomass basis better reflect the potential productivity at tight saturation than if expressed on a pigment basis. Gracilaria sp. grew better under non–filtered fluorescent and greenish than under reddish and blue–enriched light of equal and sub–optimal photon, fluence rate. However, the pigment relations of the algae did not change in a direction complementary to the light composition at which they grew. This, together with the relatively higher photosynthetic rates under reddish and blueish light for plants previously grown under reddish and blueish light, suggests that adaptations to variouslight spectra are based on mechanisms different from complementary chromatic adaptation of the pigments.     

LIGHT‐INDUCED MOTILE RESPONSES OF THE ESTUARINE BENTHIC DIATOMS NAVICULA PERMINUTA AND CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)1

Motility of estuarine epipelic (mud‐inhabiting) diatoms is an important adaptation to living in biofilms present within fine sediments. Motility allows cells to migrate within the photic zone in response to a wide range of environmental stimuli. The motile responses of two species of benthic diatoms to photon fluence rates and spectral quality were investigated. Cultures of Navicula perminuta (Grunow) in van Heurck and Cylindrotheca closterium (Ehrenb.) J. C. Lewin et Reimann both exhibited photoaccumulation at ～200 μmol · m^?2 · s^?1 and photodispersal from photon flux densities (PFDs) of ～15 μmol · m^?2 · s^?1. Photokinesis (changing cell speed) contributed toward photodispersal for both species, and red light (λ = 681–691 nm) was most effective at inducing this process. N. perminuta showed a phototactic (directional) response, with active movement in response to a light gradient. Although this response was exhibited in white light, these directional responses were only elicited by wavelengths from 430 to 510 nm. In contrast, C. closterium did not exhibit phototaxis under any light conditions used in this study. Motile benthic diatoms thus exhibit complex and sophisticated responses to light quantity and quality, involving combinations of photokinesis and phototaxis, which can contribute toward explaining the patterns of large‐scale cell movements observed in natural estuarine biofilms.     

LIGHT REQUIREMENTS FOR MONOSPORE GERMINATION IN BANGIA ATROPURPUREA (RHODOPHYTA)1

Monospore germination, in Bangia atropurpurea (Roth) C. Ag. [= B. fuscopurpurea (Dillw.) Lyngb.] is light-dependent. In white light, the percent germination increases with increasing photon fluence rate until the response is saturated at 35 μmol · m^?2· s^?1. At a saturating photon fluence rate in an 18:6 h L:D cycle, 9 days are required for maximum germination. Green light is the most effective spectral region for monospore germination, although the process can occur in red and blue light if sufficiently high photon fluence rates are provided. Monospore germination and photosynthetic oxygen evolution are completely inhibited by DCMU at a concentration of 1 × 10^?6 M. Germination is reduced in a low CO₂ atmosphere and does not occur in the dark when glucose, maltose or inositol are supplied. It is concluded that photosynthesis is required for monospore germination.     

Diaphototaxis and gravitaxis in a freshwater Cryptomonas

Phototaxis and gravitaxis are characterized in a freshwater species of the flagellate Cryptomonas. The phototactic orientation in this limnetic species is unusual and differs from all other Cryptomonas species studied so far: At both low (< or = 1O W m-2) and higher fluence rates it orients perpendicular to the light beam (diaphototaxis) while another freshwater Cryptomonas species (strain CR-1) is restricted to positive phototaxis and the marine species, C. maculata, shows both a positive and a more pronounced negative phototaxis. The mechanism of light direction detection seems to depend on a periodic shading or irradiation mechanism as confirmed by the disturbance of phototaxis in the presence of high viscosity media. In addition, this freshwater species possesses a rather pronounced negative gravitaxis which is only partially modified by phototaxis. The ecological consequences of this behavior are discussed.     

Phototropism and polarotropism of primary chloronemata of the moss Physcomitrella patens: responses of mutant strains

The phototropic and polarotropic responses of primary chloronemata grown from germinated minated spores of three mutant strains of the moss, Physcomitrella patens, have been studied and compared with those of the wild-type. The mutants and wild-type show the same qualitative tropic responses but differ with respect to the light conditions under which they are expressed. In both the wild-type and mutants the responses are controlled by phytochrome. In monochromatic red light, at low fluence rates, wild-type primary chloronemata grow positively phototropically in unidirectional light or perpendicular to the electrical vector (E) in polarised light; at high fluence rates growth in unidirectional light is lateral to the incident light or, in polarised light, parallel to E. The mutants, however, show only the lateral phototropic or parallel polarotropic responses at all fluence rates of red light tested. In far-red light, the wild-type primary chloronemata adopt a positive phototropic or a perpendicular polarotropic response; the mutants show the same responses but in a lower percentage of filaments. These results and those at other wavelengths indicate either that the mutants are impaired in their ability to adopt the positive phototropic and perpendicular polarotropic responses or that in the mutants the transition between the “low light” (positive phototropic-perpendicular polarotropic) and the “high light” (lateral phototropic-parallel polarotropic) responses is shifted to a lower photon fluence rate. Possible explanations of this phenotypic difference are discussed.     

The involvement of a protein kinase in phototaxis and gravitaxis of <Emphasis Type="Italic">Euglena gracilis</Emphasis>

The unicellular flagellate Euglena gracilis shows positive phototaxis at low-light intensities (<10 W/m²) and a negative one at higher irradiances (>10 W/m²). Phototaxis is based on blue light-activated adenylyl cyclases, which produce cAMP upon irradiation. In the absence of light the cells swim upward in the water column (negative gravitaxis). The results of sounding rocket campaigns and of a large number of ground experiments led to the following model of signal perception and transduction in gravitaxis of E. gracilis: The body of the cell is heavier than the surrounding medium, sediments and thereby exerts a force onto the lower membrane. Upon deviation from a vertical swimming path mechano-sensitive ion channels are activated. Calcium is gated inwards which leads to an increase in the intracellular calcium concentration and causes a change of the membrane potential. After influx, calcium activates one of several calmodulins found in Euglena, which in turn activates an adenylyl cyclase (different from the one involved in phototaxis) to produce cAMP from ATP. One further element in the sensory transduction chain of both phototaxis and gravitaxis is a specific protein kinase A. We found five different protein kinases A in E. gracilis. The blockage of only one of these (PK.4, accession No. EU935859) by means of RNAi inhibited both phototaxis and gravitaxis, while inhibition of the other four affected neither phototaxis nor gravitaxis. It is assumed that cAMP directly activates this protein kinase A which may in turn phosphorylate a protein involved in the flagellar beating mechanism.     

Phycomyces: Phototropism and light-growth response to pulse stimuli

The relationship between phototropism and the light-growth response of Phycomyces blakesleeanus (Burgeff) sporangiophores was investigated. After dark adaptation, stage-IVb sporangiophores were exposed to short pulses of unilateral light at 450 nm wavelength. The sporangiophores show a complex reaction to pulses of 30 s duration: maximal positive bending at 3·10^-4 and 10^-1 J m^-2, but negative bending at 30 J m^-2. The fluence dependence for the light-growth response also is complex, but in a different way than for phototropism; the first maximal response occurs at 1.8·10^-3 J m^-2 with a lesser maximum at 30 J m^-2. A hypertropic mutant, L85 (madH), lacks the negative phototropism at 30 J m^-2 but gives results otherwise similar to the wild type. The reciprocity rule was tested for several combinations of fluence rates and pulse durations that ranged from 1 ms to 30 s. Near the threshold fluence (3·10^-5 J m^-2), both responses increase for pulse durations below 67 ms and both have an optimum at 2 ms. At a fluence of 2.4·10^-3 J m^-2, both responses decrease for pulse durations below 67 ms. The hypertropic mutant (madH), investigated for low fluence only, gave similar results. In both strains, the time courses for phototropism and light-growth response, after single short pulses of various durations, show no clear correlation. These results imply that phototropism cannot be caused by linear superposition of localized light-growth responses; rather, they point to redistribution of growth substances as the cause of phototropism.     

Light quality growth promotion of Spiraea nipponica: the influence of a low photon fluence rate and transfer time to a higher fluence rate

Combinations of different light quality and fluence exposure times were investigated for their effects on in vitro growth of the woody plant Spiraea nipponica. An interaction was demonstrated between different levels of benzyladenine (BA) used for in vitro propagation and the specific light regimes investigated. This relationship was affected by the length of exposure to either white or red/FR light and the time of transfer from one fluence rate to another. In all instances exposure to red/FR light resulted in more extensive growth than under white light. Thus explants cultured under 0.25 and 0.4 mg l^-1 of BA exhibited high shoot proliferation rates when transferred, after 4 weeks of low photon fluence red/FR light, to higher fluence white light for a further week. The proliferation rates obtained were higher than any white light treatment including that with the highest BA level of 0.5 mg l^-1. In addition, the combination of red/FR light exposure with a white light stage of higher fluence improved proliferation at lower exogenous BA levels.     

Chloroplast movements in fern leaves: correlation of movement dynamics and environmental flexibility of the species

The movements of chloroplasts in response to varying levels and wavelengths of incident light were investigated in leaves of four fern species: Adiantum capillus-veneris, Adiantum caudatum, Adiantum diaphanum and Pteris cretica. In all of the species studied blue light induced chloroplast redistribution resulting in face and profile patterns that were typical of low and high fluence rates, respectively. Fluence rate response characteristics and the kinetics of transmission changes accompanying these blue-light-induced movements were similar to those observed in the leaves of higher plants. Only in A. capillus-veneris was the distribution of chloroplasts affected by red light. The response was of the weak-light type, irrespective of the light intensity. The most effective fluence rate for red light was found to be below 7·2 μmol m^–2 s^–1 (1 W m^–2). The effect of red light was far-red reversible, indicating phytochrome involvement. Chloroplast responses were more dynamic in A. capillus-veneris and P. cretica, the two species that exhibited higher environmental flexibility.     

PHYSIOLOGICAL ECOTYPES IN THE MARINE ALGA BOSTRYCHIA RADICANS (CERAMIALES,RHODOPHYTA) FROM THE EAST COAST OF THE U.S.A1

The comparative ecophysiology of nine culture isolates of the eulittoral red alga Bostrychia radicans (Montagne) Montague collected at sites from seven states along the east coast of the U.S.A. was investigated. The growth response in relation to different salinity and light conditions as well as photosynthesis-irradiance curves were studied. In addition, the effect of salt treatment on the content of the isomeric polyols d -sorbitol and d -dulcitol was also studied. All isolates grew between salinities of 5.3 and 70 ppt but with quite different optima and maxima. The isolates were all adapted to low light levels, i.e. growth was already recorded at 2.5 μmol photons·m^?2·s^?1, and growth rates peaked between 40 and 60 μmol photons·m^?2·s_-1. These low-light requirements were also reflected by the photosynthesis-irradiance curves: all plants had low light compensation points (2.5–9.7 μmol photons ·m^?2·^?1) and low photon fluence rates for initial saturation of photosynthesis (38.1–84.7 μmol photons·m^?2·s^?1, indicating that these isolates are “shade-adapted.” Isolates from Florida and Georgia synthesized and accumulated both the osmolytes d -sorbitol and d -dulcitol in increasing salinities, whereas only d -sorbitol was present in plants from North Carolina north to Connecticut. d -sorbitol was always strongly involved in osmotic acclimation. In various isolates from the same location in South Carolina, both polyol patterns were found, i.e. d -sorbitol plus d -dulcitol and d -sorbitol only. All data indicate that B. radicans exhibits a broad salinity tolerance and a low-light preference, which explain the successful colonization of this alga on various intertidal and shaded substrates. The data also clearly indicate intraspecific differences among the nine isolates, which is interpreted as development of different physiological ecotypes.     

Fluence rate and wavelength dependence of photobleaching in the cyanobacterium Anabaena variabilis

The fluence rate dependence of the photobleaching in the cyanobacterium Anabaena variabilis was studied under physiological conditions. According to the in-vivo absorption spectra measured every day during the 5 d exposition the phycobiliproteins are more sensitive to high fluence rates than chlorophyll a. The carotenoids are least sensitive, so that a relative, but not an absolute increase in the carotenoid content occurred. At very high fluence rates exceeding about 50 Wm^-2 white light the organisms were photokilled after 5 d of irradiation. Measurements of the nitrate concentrations during the experiments have shown that nitrate was not the limiting factor in these experiments. Analysis of the photobleaching kinetics at 13.5 Wm^-2 white light revealed that after about 8 d the contents of all the pigments studied have reached a new, constant level. After exposure of the photobleached cyanobacteria to low irradiances repigmentation occurred. Thus, photobleaching is a light adaptation process and not simply a photodamage phenomenon. Studying the wavelength dependence of photobleaching at a constant photon fluence rate of 4·10^-8 mol cm^-2 s^-1 we found that the photobleaching of both phycobiliproteins and chlorophyll a was exclusively caused by wavelengths absorbed by the phycobiliproteins, mainly phycoerythrocaynin, and red light absorbed by short wavelength chlorophyll. Wavelengths <520 nm were ineffective.     

Adaptation to warming but not cooling at slow rates of stimulus change in thermal threshold measurements

The relationship between thermal detection threshold and rate of temperature change of the thermal stimulus when slow (<1°C?s^?1) rates of change are employed was investigated. Using both the reaction time (RT) inclusive Method of Limits and RT exclusive Method of Levels healthy volunteers had warming (WDT) and cooling detection thresholds (CDT) measured at four different rates of temperature change (0.3, 0.5, 0.7 and 1.0°C?s^?1) from the thenar and/or mental regions using a contact thermode. With the Method of Limits, CDT increased linearly with rate of temperature change suggesting increments were due to RT artefacts. This was further supported by threshold assessment with the Method of Levels which showed CDT were unaffected by the rate of change in the RT exclusive method (P?>?0.1). In contrast, WDT did not increase linearly with rate of stimulus temperature change when the Method of Limits was used and threshold assessment with the Method of Levels showed WDT assessed using a 0.3°C?s^?1 ramp rate were significantly higher than those measured with a 1°C?s^?1 rate of change (P?<?0.05). This study indicates that adaptation to a warming stimulus can occur at faster rates of stimulus change than previously anticipated and identifies differences in warming and cooling pathways in sensitivity to adaptation.     

Separation and properties of prestalk and prespore cells of Dictyostelium discoideum

We describe a method of separating prestalk and prespore cells of Dictyostelium discoideum slugs using a self-generating Percoll gradient. This method gives quantitative recovery of cells and good purity. Separated prestalk and prespore cells possess different levels of the enzymes UDP galactose :polysaccharide transferase, cAMP phosphodiesterase and glycogen phosphorylase. We have used this method, as well as mechanical dissection of slugs, to examine the fate of separated prestalk and prespore cells in Dictyostelium strains that are able to give rise to mature stalk and spore cells in cell monolayers. The results from such experiments provide direct evidence that prestalk and prespore cells from the migrating slug stage are programmed to differentiate into stalk and spore cells respectively.     

The influence of light on copper‐limited growth of an oceanic diatom,Thalassiosira oceanica (Coscinodiscophyceae)

Thalassiosira oceanica (CCMP 1005) was grown over a range of copper concentrations at saturating and subsaturating irradiance to test the hypothesis that Cu and light were interacting essential resources. Growth was a hyperbolic function of irradiance in Cu‐replete medium (263 fmol Cu′ · L^?1) with maximum rates achieved at 200 μmol photons · m^?2 · s^?1. Lowering the Cu concentration at this irradiance to 30.8 fmol Cu′ · L^?1 decreased cellular Cu quota by 7‐fold and reduced growth rate by 50%. Copper‐deficient cells had significantly slower (P < 0.0001) rates of maximum, relative photosynthetic electron transport (rETR_max) than Cu‐sufficient cells, consistent with the role of Cu in photosynthesis in this diatom. In low‐Cu medium (30.8 fmol Cu′ · L^?1), growth rate was best described as a positive, linear function of irradiance and reached the maximum value measured in Cu‐replete cells when irradiance increased to 400 μmol photons · m^?2 · s^?1. Thus, at high light, low‐Cu concentration was no longer limiting to growth: Cu concentration and light interacted strongly to affect growth rate of T. oceanica (P < 0.0001). Relative ETR_max and Cu quota of cells grown at low Cu also increased at 400 μmol photons · m^?2 · s^?1 to levels measured in Cu‐replete cells. Steady‐state uptake rates of Cu‐deficient and sufficient cells were light‐dependent, suggesting that faster growth of T. oceanica under high light and low Cu was a result of light‐stimulated Cu uptake.     

DEPRESSION OF MASTIGOCLADUS LAMINOSUS (CYANOPHYTA) NITROGENASE ACTIVITY UNDER NORMAL SUNLIGHT INTENSITIES1

The effect of light intensity (PAR) on the nitrogenase activity of Mastigocladus laminosus Cohn was studied by the acetylene reduction technique. Benthic mat from a thermal stream, Hot River, in Yellowstone National Park was used in both experimental and in situ incubations. This hot spring maintained a mean pH of 7.0, was essentially isothermal (ca. 50°C), and had virtually no upstream to downstream physicochemical gradients (P > 0.05). Two surveys of the stream showed that nitrogenase of the M. laminosus mat was significantly more active (P > 0.02) under low light intensities than under high intensities, 252 and 712 μE · m^?2· s^?1, respectively. Maximum activity of Hot River Mastigocladus (268 nmol C₂H₄· mg Chl a^?1· h^?1) occurred at 50% full midday light intensities; the rates at low light (mean = 247 nmol C₂H₄· mg Chl a^?1· h^?1) were significantly (P > 0.001) greater than those at high light (mean = 106). The results indicate that M. laminosus nitrogenase activity is low light adapted and suggest that the temporal pattern for nitrogen fixation might be significantly different from that of thermophilic Calothrix.     

The relationship between the redox state of Q A and photosynthesis in leaves at various carbon-dioxide,oxygen and light regimes

The response of chlorophyll fluorescence elicited by a low-fluence-rate modulated measuring beam to actinic light and to superimposed 1-s pulses from a high-fluence-rate light source was used to measure the redox state of the primary acceptor Q _A of photosystem II in leaves which were photosynthesizing under steady-state conditions. The leaves were exposed to various O₂ and CO₂ concentrations and to different energy fluence rates of actinic light to assess the relationship between rates of photosynthesis and the redox state of Q _A. Both at low and high fluence rates, the redox state of Q _A was little altered when the CO₂ concentration was reduced from saturation to about 600 l·l^-1 although photosynthesis was decreased particularly at high fluence rates. Upon further reduction in CO₂ content the amount of reduced Q _A increased appreciably even at low fluence rates where light limited CO₂ reduction. Both in the presence and in the absence of CO₂, a more reduced Q _A was observed when the O₂ concentration was below 2%. Q _A was almost fully reduced when leaves were exposed to high fluence rates under nitrogen. Even at low fluence rates, Q _A was more reduced in shade leaves of Asarum europaeum and Fagus sylvatica than in leaves of Helianthus annuus and Fagus sylvatica grown under high light. Also, in shade leaves the redox state of Q _A changed more during a transition from air containing 350 l·l^-1 CO₂ to CO₂-free air than in sun leaves. The results are discussed with respect to the energy status and the CO₂-fixation rate of the leaves.Abbreviations and symbols L 1,2 first and second actinic light beam - Q _A primary acceptor of photosystem II - q _Q Q-quenching