Cd has pleiotropic effects on plant physiology and in particular on photosynthesis. It has not been established yet if Cd alters the functioning of the xanthophyll cycle. To answer this question, an exponentially growing culture of the marine diatom Phaeodactylum tricornutum was incubated with Cd (20 mg/l) for 24 h and irradiated with a light activating the xanthophyll cycle, which in diatoms, consists of the reversible deepoxidation of diadinoxanthin to diatoxanthin. The measurements show that the deepoxidation step is not influenced by Cd. In contrast, the Cd concentration used sharply inhibits the epoxidation of diatoxanthin to diadinoxanthin. ß
Pronounced aggregation of the photosystem I1 light-harvesting complex (LHC 11) was observed in low-lightgrown tobacco plants stressed with a strong CO, deficit for 2-3 days. The LHC I1 aggregates showed a typical band at 697-700 n m (F,99) in low-temperature emission spectra. Its excitation spectrum corresponded to that of detergent-solubilized LHC 11. Formation of F699 in stressed plants was not reversed in the dark and leaves did not contain any zeaxanthin showing that neither a light-induced transthylakoid pH gradient nor zeaxanthin was required for LHC I1 aggregation. The C02-stressed plants showed clear signs of photodamage: depression of the potential yield of photosystem I1 photochemistry (FJ F,) by 50-70% and a decline in chlorophyll content by 10-15%. Therefore, we propose that the photodamage to the photosynthetic apparatus is the cause of the LHC I1 aggregation in plants. The Fsg9 exhibited a reversible decrease of its intensity upon irradiation of leaves with intensive light. There was no or only slight decrease around 700 nm in unstressed plants. The nonphotochemical quenching of chlorophyll fluorescence showed the opposite relation, being higher before than after the strong C 0 2 deficit. This discrepancy was likely related to the different LHC 11 aggregation state in control and stressed plants.
The UV light (337 nm) induced blue-green fluorescence emission of green leaves is characterized at room temperature (298 K) by a maximum near 450 nm (blue region) and a shoulder near 525 nm (green region) and was here also studied at 77K. At liquid nitrogen temperature (77 K) the blue (F450) and green fluorescence (F525) are much enhanced as is the red chlorophyll fluorescence near 735 nm. During development of green tobacco leaves the blue fluorescence F450 (77 K)is shifted towards longer wavelengths from about 410 nm to 450 nm. The isolated leaf epidermis of tobacco showed only slight fluorescence emission with a maximum near 410nm. The green fluorescence F525 was found to mainly originate from the mesophyll of the leaf, its intensity increased when the epidermis was removed. The red chlorophyll fluorescence emission was also enhanced when the epidermis was stripped off; this considerably changed the blue/red fluorescence ratios F450/F690 and F450/F735. The epidermis, with its cell wall and W-light-absorbing substances in its vacuole, plays the role of a barrier for the exciting Wlight. In contrast to intact and homogenized leaves, isolated intact chloroplasts and thylakoid membranes did not exhibit a blue-green fluorescence emission. Abbreviations and Symbols F450:blue fluorescence emission at 450 nm F525:green fluorescence emission at 525 nm F685, chlorophyll fluorescence emission at 685, F695, F735: 695 and 735 nm, respectively Bot. Acta 105 (1992) 435-440
Changes in excitonic interactions of photosystem II (PSII) reaction centre (RC) pigments upon light-induced oxidation of primary donor (P680) or reduction of primary acceptor (pheophytin (Pheo)) were analysed using circular dichroism (CD). The CD spectrum of PSII RC shows positive bands at 417, 435 and 681 and negative bands at 447 and 664 nm. Oxidation of the primary donor by illuminating the sample in the presence of silicomolybdate resulted in nearly symmetric decrease of CD amplitudes at 664 and 684 nm. In the Soret region, the maximum bleaching of CD signal was detected at 449 and 440 nm. Accumulation of reduced Pheo in the presence of dithionite brought about much lower changes in CD amplitudes than P680 oxidation. In this case, only a small asymmetric bleaching at 680 and 668 nm in the red region and a bleaching at 445, 435 and 416 nm in the Soret region has been detected. Therefore, we suppose that the contribution of the Pheo of the primary acceptor to the total CD signal of RC is negligible. In contrast to the oxidation of primary donor, the light-induced change in the CD spectrum upon primary acceptor reduction was strongly temperature-dependent. The reversible CD bleaching was completely inhibited below 200 K, although the reduced Pheo was accumulated even at a temperature of 77 K. Since the temperature does not influence the excitonic interaction, the temperature dependence of the CD changes upon Pheo reduction does not support the model of Pheo excitonically interacting with the other chlorophylls (Chl) of the RC. We propose that Pheo should not be considered as a part of a multimer model.
A multichannel kinetic spectrophotometer-fluorimeter with pulsed measuring beam and differential optics has been constructed for measurements of light-induced absorbance and fluorescence yield changes in isolated chlorophyll-proteins, thylakoids and intact cells including algae and photosynthetic bacteria. The measuring beam, provided by a short (2 micros) pulse from a xenon flash lamp, is divided into a sample and reference channel by a broad band beam splitter. The spectrum in each channel is analyzed separately by a photodiode array. The use of flash measuring beam and differential detection yields high signal-to-noise ratio (noise level of 2 x 10(-4) in absorbance units per single flash) with negligible actinic effect. The instrument covers a spectral range between 300 and 1050 nm with a spectral resolution of 2.1, 6.4 or 12.8 nm dependent on the type of grating used. The optical design of the instrument enables measuring of the difference spectra during an actinic irradiation of samples with continuous light and/or saturation flashes. The time resolution of the spectrophotometer is limited by the length of Xe flash lamp pulses to 2 micros.
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