The composition and function of thylakoid membranes from pea plants grown under white or green light with or without far-red light

Pea plants ( Pisum sativum L. ev. Greenfeast) were grown for 2 to 3 weeks in while (˜ 50 μmol photons m⁻² s⁻¹; 400–700 nm) or green (˜ 30 μmol photons m⁻² s ⁻¹ 400–700 nm) light (16 h day/8 h night), with or without far-red light. Supplementary far-red light decreased leaf area and increased internodal length in both white and green light, demonstrating that phytochrome influenced leaf size and plant growth. However, there was no effect of far-red light on chlorophyll a /chlorophyll b ratios, chlorophyll-protein composition, the stoichiometry of electron transport complexes or photosynthetic function of isolated thylakoids. These results suggest that phytochrome is ineffective in modulating the composition and function of thylakoids in pea plants grown at low irradiance. One possible explanation of the ineffectiveness of phytochrome on thylakoids is discussed in terms of the drastic attenuation of red relative to far-red light in green tissue.     

Effects of oxidative stress on chlorophyll biosynthesis in cucumber (Cucumis sativus) cotyledons

We conducted a series of experiments to assess the effects of oxidative stress on chlorophyll biosynthesis in the vascular plant Cucumis sativus (cucumber). Specifically, cucumber cotyledons were treated with 100 μ M methyl viologen (MV) and subsequently exposed to dark (0 μE m⁻² s⁻¹), low light (40–45 μE m⁻² s⁻¹), or high light (1500–1600 μE m⁻² s⁻¹). Following treatment, extracts of these samples were subjected to high-performance liquid chromatography (HPLC) to quantitate the accumulation of chlorophyll biosynthetic pathway intermediates. The results of these analyses revealed significant accumulation of Mg-protoporphyrin IX monomethyl ester (Mg-proto IX ME) in green (14-h illuminated) as well as in etiolated cotyledons with MV treatment. These data suggest that MV-induced oxidative stress may have inhibited Mg-proto IX ME cyclase activity. Upon exposure to high light, in the presence or absence of MV, both green and etiolated cotyledons predominantly accumulated protoporphyrin IX (Proto IX). These elevated levels of Proto IX might be attributable to attenuated activity of any or all of the following enzymes: Mg-chelatase, Fe-chelatase and protoporphyrinogen IX oxidase. We also observed that MV-induced oxidative stress impacts on chlorophyll biosynthesis to a greater extent than on photosystem II. These results demonstrate that oxidative stress impedes key steps in chlorophyll biosynthesis by either directly or indirectly inhibiting the activity of these enzymes.     

Light effects on in vitro adventitious root formation in axillary shoots of mature Prunus serotina

Light effects on in vitro adventitious root formation in axillary shoots of a 95-year-old black cherry ( Prunus serotina Ehrh.) were examined using microcuttings derived from cultured vegetative buds. Three studies were performed: 1) complete darkness and 4 levels of continuous white light irradiance were tested at 70, 278, 555 and 833 μmol m⁻² s⁻¹; 2) white, red, yellow and blue light were tested to assess the importance of spectral quality; and 3) the effect of blue light at intensities of 7,15, 22 and 30 μmol m⁻² s⁻¹ was also studied, Measurements included rooting percentage, total number of roots per shoot, and shoot and root dry weight. There was a strong negative effect of white light intensity upon root formation. Blue light between 15 and 22 μmol m⁻²: s⁻¹ significantly retarded root formation and completely inhibited it at 36 μmol m⁻² s⁻¹. Shoots treated with yellow light exhibited the highest rooting percentage, mean number of roots per shoot, and root dry weight.     

PHOTOSYNTHETIC RESPONSE OF STREAM PERIPHYTON TO FLUCTUATING LIGHT REGIMES

  An experiment was conducted on intact algal assemblages of stream periphyton to test their response to fluctuating and constant light regimes having the same mean intensity. The light regimes (in μmol·m⁻²·s⁻¹) were constant light at 100, light fluctuating between 50 and 150 with a period of 5 min, and light fluctuating between 10 and 460 with periods of either 4:1 or 8:2 min. Compared to the rates measured under 100 in μmol·m⁻²·s⁻¹ constant light conditions, fluctuations ranging between 50 and 150 in μmol·m⁻²·s⁻¹ with a 5-min period produced a 23% greater rate of photosynthesis. Conversely, fluctuations between 10 and 460 in μmol·m⁻²·s⁻¹ led to a 59%–74% decrease in photosynthetic activity. Detailed examination of periphytic algal responses to fluctuating light revealed that higher light intensities produced steeper photosynthesis/time slopes, but it was the combined interaction with lower light intensity that ultimately determined overall photosynthetic rate for a given light regime. This study offers compelling evidence that variable light regimes have important consequences for algal photosynthesis in natural streams.     

Characteristics of the light-induced generator potentials in the liverwort Conocephalum conicum

Light-induced generator potentials were examined in the liverwort Conocephalum conicum L. by applying sub-threshold light stimuli (400–750 nm). The fluenceresponse dependence was determined for various wavelengths. At low values of quantum flux density (less than 1 μmol m⁻² s⁻¹) the plant reacts to the light pulses with hyperpolarization which, with a rise in the pulse intensity, is masked by the depolarization of the transmembrane potential. The action spectrum of depolarization shares many features with the absorption specturm of photosynthetic pigments. Application of inhibitors and electron donors showed a predominant role of photosystem II in the formation of the generator potential. No significant Emerson enhancement was observed after simultaneous illumination with light beams of 650 and 700 nm.     

Mechanism of copper-enhanced photoinhibition in thylakoid membranes

The effect of copper on photoinhibition of photosystem II (PSII) in vitro was studied in bean ( Phaseolus vulgaris L. cv. Dufrix) and pumpkin ( Cucurbita pepo L.) thylakoids. The thylakoids were illuminated at 200–2 000 μmol photons m⁻² s⁻¹ in the presence of 70–1 830 added Cu²⁺ ions per PSII. Three lines of evidence show that the irreversible damage of PSII caused by illumination of thylakoids in the presence of Cu²⁺ was mainly due to donor-side photoinhibition resulting from inhibition of the PSII donor side by Cu²⁺. First, addition of an artificial electron donor partially restored PSII activity of thylakoids that had been illuminated in the presence of Cu²⁺. Second, already moderate light was enough to cause rapid inhibition of PSII, and the inhibition could be saturated by light. Third, the extrinsic polypeptides of the oxygen-evolving complex were found to become oxidized by the combined effect of Cu²⁺ and light. The presence of oxygen was not necessary for the copper-induced enhancement of photoinhibition of PSII. When the illumination was prolonged, copper caused a gradual collapse of the thylakoid structure by increasing degradation of thylakoid proteins.     

Effect of ultraviolet radiation on accumulation and leakage of 86Rb+ in guard cells of Vicia faba

The effects of UV-C (254 nm), UV-A (365 nm) and broad-band UV (280–380 nm) on guard cells of Vicia faba L. cv. Long Pod were investigated in the presence of white light (450 μmol m⁻² s⁻¹). UV-C (7 μmol m⁻² s⁻¹) was found to cause leakage of ⁸⁶Rb⁺ from guard cells, while UV-A (0.3 μmol m⁻² s⁻¹) stimulated increased uptake in these cells. A relatively small stimulatory effect was observed by broad-band UV (3 μmol m⁻² s⁻¹) during the first 30 min of irradiation with an apparent equilibration of influx and efflux thereafter. Leakage of ⁸⁶Rb⁺ from guard cells continued despite the removal of UV-C and an increase in the amount of white light from 450 to 1500 μmol m⁻² s⁻¹, suggesting that membranes were irreversibly damaged. Irradiation of guard cells with UV-C for 30, 45 and 90 min indicated that these cells began to be affected already by 30 min UV-C irradiation.     

CIRCADIAN GROWTH IN PORPHYRA UMBILICALIS (RHODOPHYTA): SPECTRAL SENSITIVITY OF THE CIRCADIAN SYSTEM

The circadian rhythm in growth of the red macroalga Porphyra umbilicalis (Linnaeus) J. Agardh was investigated under different spectral light conditions in laboratory-grown thalli. A free-running rhythm was observed in constant green or red light at irradiances of 2.5 to 20 μmol photons·m⁻²·s⁻¹, whereas arhythmicity occurred in constant blue light at 6–20 μmol photons·m⁻²·s⁻¹. The circadian oscillator controlling growth rhythmicity in Porphyra uses most of the visible sunlight spectrum and possibly multiple photoreceptors with a high sensitivity for blue light and a lower sensitivity for red light. This was inferred from three experimental results: (1) The free-running period, τ, of the growth rhythm decreased with increasing irradiance, from approximately 25 h at 2.5 μmol photons·m⁻²·s⁻¹ to 22 h at 20 μmol photons·m⁻²·s⁻¹ in red or green light, (2) Dark pulses of 3 h duration, interrupting otherwise continuous green or red light, caused advances during the subjective day and delays during the subjective night; the circadian oscillator in Porphyra can discriminate darkness from green or red light, and (3) Low-irradiance blue light pulses (2.5 μmol photons·m⁻²·s⁻¹) shifted the growth rhythm in red light of higher irradiance (e.g. 10 μmol photons·m⁻²·s⁻¹), and a strong, high amplitude, type 0 phase response curve was obtained that is usually observed with light pulses shifting a circadian rhythm in otherwise continuous darkness.     

Photosystem I acceptor side limitation is a prerequisite for the reversible decrease in the maximum extent of P700 oxidation after short-term chilling in the light in four plant species with different chilling sensitivities

Changes in the extent of P700 oxidation (P700⁺) were investigated after chilling of barley, rice, pumpkin, and cucumber leaf segments at 4°C for 1 h under light with various photon flux densities. At 50 µmol photons m⁻² s⁻¹, the decrease in P700⁺ was observed only in cucumber, but at 150 µmol photons m⁻² s⁻¹, it was found in all plants except barley, revealing their expected chilling sensitivities. However, the decrease in P700⁺ by this short-term chilling was reversible in the presence of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea or methyl viologen, and it did not show any causal relationship with the decrease in the electron transfer rate nor with the down-regulation of photosystem II through the accumulation of zeaxanthin and the development of non-photochemical quenching. These results led to the suggestion that photosystem I (PSI) acceptor side limitation is a prerequisite for the decrease of P700⁺. Furthermore, PSI acceptor side limitation could be mainly due to limitation of electron-sink pathways such as CO₂ assimilation and ascorbate–glutathione cycle, because treatment with glycolaldehyde which inhibits the former pathway, and with KCN which inhibits both pathways, decreased P700⁺ by 20–30% in barley leaves after chilling in the light.     

Effect of light quality (blue, red) and fluence rate on the synthesis of pigments and pigment-proteins in maize and black pine mesophyll chloroplasts

Maize ( Zea mays L. hybrid ZP-704) and black pine ( Pinus nigra Arn.) were grown for five days at low fluence rate (0.4–4.0, μmol m^–2 s⁻¹) in blue or red light. Compared to red light of the same fluence rate, blue light effects in maize were repressive for the accumulation of Chita, b , carotenoids and light-harvesting complex-2 (LHC-2) proteins. The maximal reduction of proteins bound to the light-harvesting complex of photosystem 2 and pigments was attained at different fluence rate levels. In black pine, blue light compared to the red of the same fluence rate level either activated or reduced accumulation of pigments and LHC proteins, the effect being dependent on its fluence rate level. At fluence less than 3.0 μmol m⁻² s⁻¹ blue light was more efficient for the synthesis of Chi a, b and carotenoids, hut for LHC-2 complexes, fluence rates between 0.4 and 1.5 [μmol m⁻² s⁻¹ were more effective. In pine the effects of the two lights on the accumulation of pigments and LHC proteins were demonstrated separately and were dependent on fluence rate level. This suggests irradianoe-controlled activation/deactivation of the photoreceptor at the level of the cell.     

The effects of cadmium on photosynthesis of Phaseolus vulgaris – a fluorescence analysis

Bean plants ( Phaseolus vulgaris L. cv. Scarlett), germinated in darkness for I week, were transferred to light (200 μmol m⁻² s⁻¹) and cultivated for I week in a complete nutrient solution. After this period, cadmium ions in the form of CdSO₄ were added at the concentrations of 0.10.20 and 50 μ M . The effects of this metal on the properties of photosystem II photochemistry were studied by means of modulated fluorescence analysis. Steady state photochemical quenching. non-photochemical quenching and terminal fluorescence were determined in control and cadmiumtreated plants. We postulate that, during short term exposure of plants to cadmium in the early stages of growth, the Calvin cycle reactions are more likely than photosystem II to be the primary target of the toxic influence of cadmium. The reduced demand for ATP and NADPH upon Calvin cycle inhibition causes a down-regulation of photosystem II photochemistry and of the yield of linear electron transport.     

Photophobic stop-response in a dinoflagellate: Modulation by preirradiation

Gyrodinium dorsum Kofoid responds photophobically to flashes of blue light. The photophobic response consists of a cessation of movement (stop-response). Without background light and after a flash fluence above 10 J m⁻², 75–85% of the cells show a stop-response, while only 50% of the cells show this response at 5 J m⁻². With a flash fluence of 5 J m⁻², background light of different wavelengths either increases (614 nm. 5.5–18.2 μmol m⁻² s⁻¹) or decreases (700 nm, 18.4–36.0 μmol m⁻² s⁻¹) the stop-response. Two hypotheses for the mechanism of the modulation by background light of the photophobic response are discussed: an effect of light on the balance of the photosynthetic system (PS I/PS II) or an effect on a phytochrome-like pigment (P_r/P_fr). This study supports the idea that a phytochrome-like pigment works in combination with a blue light-absorbing pigment. It was also found that cells of Gyrodinium dorsum cultured in red light (39.8 μmol m⁻²) had a higher absorption in the red region of the absorption spectra than those cultured in white light (92.7 μmol m⁻²).     

Influence of irradiance on in vitro growth and proliferation of Vaccinium corymbosum (highbush blueberry) and subsequent rooting in vivo

The effects of light on in vitro proliferation and subsequent in vivo rooting and acclimatisation of Vaccinium corymbosum were investigated. The shoots were exposed in vitro to different irradiances (total radiation ranging from 55 to 240 μmol m⁻² s⁻¹) for 7 to 60 days. In vitro growth and proliferation and the possible consequences on in vivo rooting were observed.
As compared to the control treatment (55 μmol m⁻² s⁻¹), higher irradiances improved proliferation and rooting ratios only with short applications (7 days). Short but high (210 μmol m⁻² s⁻¹) exposures applied at the end of the proliferation phase increased in vivo growth and rooting of the shoots. The shoots treated with strong light for longer times (14 and 28 days) showed both inhibition of growth and red colour of leaves and sprouts, and were less vigorous when transferred in vivo.     

Salt induced responses of chloroplast activities in species of differing salt tolerance. Photosynthetic electron transport in Aster tripolium and Pisum sativum

A comparative study was made of the effects of high concentrations of NaCl, KCl and MgCl₂ on two electron transport reactions of thylakoids isolated from a mesophyte, Pisum sativum and a halophyte, Aster tripolium . The rate of photosystem I mediated electron transport from reduced N, N, N', N'-tetramethyl- p -phenylenediamine (TMPD) to methyl viologen was determined polarographically, and photosystem II mediated electron flow from water to 2,6-dichlorophenolindophenol (DCPIP) was monitored spectrophotometrically. The response of photosystem II to increasing in vitro salt concentrations was similar for thylakoids isolated from both A. tripolium and P. sativum , but differences in the response of photosystem I to salinity changes were observed for the two species. Increasing NaCl, KCl and MgCl₂ concentrations produced similar patterns of response of photosystem I activity in P. sativum thylakoids, whilst for A. tripolium KCl induced a completely different response pattern compared to NaCl and MgCl₂. The salinity of the culture medium in which A. tripolium was grown also had an effect on both the absolute in vitro activities of photosystems I and II and their response to changes in salt concentration of the reaction media.     

Photoinhibition of photosynthesis and growth responses at different light levels in psbA gene mutants of the cyanobacterium Synechococcus

Photoinhibition of photosynthesis and growth responses at diffrent light levels (10, 120 and 250 μmol m⁻² s⁻¹) were studied in psbA gene mutants R2S2C3 ( psbAI gene present) and R2K1 ( psbAIIIpsbAIII genes present) of the cyanobacterium Synechococcus sp . PCC 7942 ( Anacystis nidulans R2). Mutant R2K1 (possessing form II of the D1 protein of photosystem II) was much more resistant to photoinhibition than the mutant R2S2C3 (possessing form I of the D1 protein). At moderate inhibitory light levels (100 to 300 μmol m⁻² s⁻¹) this was largely ascribed to an increased rsistance of the photosystem II reaction cetres possessing form II of the D1 protein. However, at higher light levels the higher resistance mutant R2K1 was assigned to a higher rate of photosystem II repair, i.e. turnover of the D1 protein. Moreover, our results support the hypothesis that photoinhibition of photosystem II and photoinhibitory induced quenching are due to separate processes. Results from growth experiments show that the R2K1 mutant has a slower growth rate than the R2S2C3 mutant but shows an increased survival under high light stress conditions. It is hypothesized that high resistance to photoinhibition, though allowing a better survival under high light, is not advantageous for optimal growth.     

Benzyladenine-induced stimulation of 5-aminolevulinic acid accumulation under various light intensities in levulinic acid-treated cotyledons of etiolated cucumber

Benzyladenine (BA) stimulated 5-aminolevulinic acid (ALA) accumulation in the presence of levulinic acid during illumination with 43 μmol m⁻² s⁻¹ light in excised etiolated cotyledons of cucumber ( Cucumis sativus L. cv. Aonagajibai). A short dark-pretreatment (6 h) with BA eliminated the lag phase of ALA accumulation. The rate of ALA accumulation during the steady-state phase in cotyledons pretreated with BA for a long period (14 h) was considerably accelerated compared to that in cotyledons pretreated with BA for 6 h. The rate of ALA accumulation during the lag phase was saturated at a very low light fluence (<1.4 μmol m⁻² s⁻¹) in both BA-pretreated and water-control cotyledons. The steady-state rate of ALA accumulation increased with increasing light fluence up to 43 μmol m⁻² s⁻¹ (parallel to that of Chl formation) in water-control cotyledons. In contrast, in cotyledons pretreated with BA for either 6 or 14 h, the steady-state rate reached a plateau at a very low light fluence. Based on the above results together with our finding that there are two components of Chl formation (M. Dei, 1984. Physiol. Plant. 62: 521–526) possible intermediate steps of Chl biosynthesis pathway affected by BA and light intensity are discussed.     

The effect of turbidity and illumination on the reaction distance and search time of the marine planktivore Gobiusculus flavescens

Both reduced illumination and increased turbidity caused a significant reduction in reaction distance of Gobiusculus flavescens . The longest reaction distance, 18.9 cm for larger prey (Calanus finmarchicus) , occurred at a light level of 80 μmol m ⁻² s ⁻¹ compared to 12.9 cm for a smaller prey (Acartia clausi) at 8 μmol m⁻² s⁻¹. Above a light saturation level of 10 μmol m⁻² s⁻¹, additional light had little influence on reaction distance. In the turbidity experiments, the longest reaction distances were measured at turbidity levels of 10–20 JTU. Prey size influenced reaction distance at all tested light levels. Search time was influenced by prey size only at low illumination. With increasing turbidity, reaction distance to a group of prey was longer than to one prey.     

Effect of light and gibberellic acid on photosynthesis during leaf senescence of alstroemeria cut flowers.

The functioning of the photosynthetic apparatus during leaf senescence was investigated in alstroemeria cut flowers by a combination of gas-exchange measurements and analysis of in vivo chlorophyll fluorescence. Chlorophyll loss in leaves of alstroemeria cut flowers is delayed by light and by a treatment of the cut flowers with gibberellic acid (GA₃). The maximal photosynthesis of the leaves was approximately 6 μmol CO₂ m⁻² s⁻¹ at I 350 μmol m⁻² s⁻¹ (PAR) which is relatively low for intact C₃ leaves. Qualitatively the gas-exchange rates followed the decline in chlorophyll content for the various treatments, i.e. light and GA₃-treatment delayed the decline in photosynthetic rates. However, when chlorophyll loss could not yet be observed in the leaves, photosynthetic rates were already strongly decreased. In vivo fluorescence measurements revealed that the decrease in CO₂ uptake is (partly) due to a decreased electron flow through photosystem II. Furthermore, analysis of the fluorescence data showed a high nonphotochemical quenching under all experimental conditions, indicating that the consumption of reducing power in the Calvin cycle is very low. The chlorophyll, remaining after 9 days incubation of leaves with GA₃ in the dark should be considered as a 'cosmetic' pigment without any function in the supply of assimilates to the flowers.     

Chloroplast structure in normal and pigment-deficient soybeans grown in continuous red or far-red light

Clark L1, a normal green soybean [ Glycine max (L.) Merrill] and Clark y₉y₉, a backross-developed isoline exhibiting pigment deficiency, were grown under continuous red (11 W m⁻² and far-red (9 W m⁻²) light. Chloroplast thylakoids from the unifoliolate leaf (9–10 days old) were isolated and analyzed for pigments, pigment-protein, membrane polypeptides, electron transport and ultrastructural differences. Chloroplasts of soybean plants grown under far-red light have decreased chlorophyll a to chlorophyll b ratio, increased light-harvesting complexes, and grana structure with few stroma-type thylakoids. Photosystem II/photosystem I ratios (PSII/PSI) are higher in far-red due to decreased synthesis of PSI reaction center and/or less antenna associated with PSI.     

Spectacular abundance of ciliates in anoxic pond water: contribution of symbiont photosynthesis to host respiratory oxygen requirements

Abstract: Very large numbers (3466 ml⁻¹) of ciliated protozoa were found living beneath the oxic-anoxic boundary in a stratified freshwater pond. Most ciliates (96%) contained symbiotic algae ( Chlorella spp.). Peak abundance was in anoxic water with almost 1 mol free CO₂ m⁻³ and a midday irradiance of 6 μmol photon m⁻² s⁻¹. Photosynthetic rate measurements of metalimnetic water indicated a light compensation point of 1.7 μmol photon m⁻² s⁻¹ which represents 0.6% of sub-surface light. We calculate that photosynthetic evolution of O₂ by symbionts is sufficient to meet the demand of the host ciliates for 13 to 14 hours each day. Each 'photosynthetic ciliate' may therefore become an aerobic island surrounded by anoxic water.